Objective: To explore and study the clinical usefulness of continuous dynamic recording of left cardiac function changes forevaluation the improvement in patients with chronic disease after 3 months of intensive control of individualized precision exercise overall manage program. Methods: From 2018 to 2021, 21 patients with chronic cardiovascular and cerebrovascular metabolic diseases mainly controlled by our team were selected to complete the cardiopulmonary exercise test (CPET) and Non-invasive synchronous cardiac function detector (N-ISCFD), electrocardiogram, radial pulse wave, jugular pulse wave and cardiogram data were continuously recorded for 50s.According to the titration results under CPET and continuous functional parameters monitoring, a holistic plan with individualized moderate exercise intensity as the core was developed for 3 months of intensive management, and then N-ISCFD data collection was repeatedafter signing the informed consent. All N-ISCFD data were analyzed in the 50s according to the optimal report mode of Fuwai Hospital and 52 cardiac functional indexes were calculated. The data before and after the enhanced control were compared and the paired T-test was used to statistically analyze the changes of groups. Results: Twenty-one patients with chronic diseases (16 male and 5 female) were (54.05±12.77,29~75) years, BMI (25.53±4.04,16.62~31.7) kg/m².Comparison with baseline,the whole group analysis: ①The body weight, BMI, systolic blood pressure and diastolic blood pressure of patients were significantly decreased(P＜0.01).②CPET Peak VO₂ was (64.93±24.22, 26.96~103.48) %Pred before enhanced control, and (85.22±30.31, 43.95~140.48) %Pred after enhanced control, and increased (35.09±27.87, 0.12~129.35) % after enhanced control compared with before enhanced control. The AT, Peak VO₂/HR, Peak Work Rate, OUEP, FVC, FEV₁, FEV3/FVC% and MVV were significantly increased (P＜0.01) and the Lowest VE/VCO₂ and VE/VCO₂ Slope were significantly decreased(P＜0.01).③Core indicators of left heart function:Ejection fraction was significantly increased from (0.60±0.12,0.40~0.88) to(0.66±0.09, 0.53~0.87)(P＜ 0.01), by (12.39±14.90,-12.32~41.11)%. The total peripheral resistance was significantly decreased from (1579.52±425.45,779.46~2409.61) G/(cm⁴·s),to(1340.44±261.49,756.05~1827.01) G/(cm⁴·s)(P＜0.01), by (12.00±17.27,37.79~28.61) %.The left stroke index, cardiac total power, ejective pressure and left ventricular end diastolic volumewere significantly improved (P＜0.05).The change analysis of each indicator for each patient is shown in the individualized analysis section of this study. Conclusion: Use CPET and continuous functional monitoring we can safely and effectively develop the overall program of individualized exercise in patients with chronic diseases. Long-term intensive management and control can safely and effectively significantly improve the cardiovascular function of patients. Continuous dynamic recording of changes in left and right cardiac functional parameters can be a simple way to supplement CPET to evaluate cardiovascular function.

Objective: To investigate the interventional effects of a new SUR2B/Kir6.1-type K_ATP Channel opener iptakalim on injury renal cells (the renal glomerular endothelial, mesangial and tubular epithelial cells) and its mechanisms. Methods: ①Experimental protocol: control: the cells were treated with with 0 mg/L uric acid for 24 h; model: the cells were treated with with 1 200 mg/L uric acid for 24 h; pretreatment with iptakalim: the cells were pretreated with 0.01,0.1,1,10,100 μmol/L iptakalim for 24 h prior to treatment with 1 200 mg/L uric acid for 24 h; pretreatment with glibenclamide: the cells were preincubated with/without 10 μmol/L glibenclamide for 1 h and then treated with 10 μmol/L iptakalim for 24 h followed by incubation with 1 200 mg/L uric acid for another 24 h. ②The cell viability was measured by MTT assay and flow cytometry; the protein expressions of Kir6.1 and SUR2B and nuclear translocation were detected by immunostaining; the protein expressions of Kir6.1 and SUR2B were determined by Western blot analysis; adhesion of mononuclear cells to endothelial cells were tested by fluorimetric assay; the content of MCP-1 was measured by enzyme linked-immunosorbent assay (ELISA). Results: The renal glomerular endothelial, mesangial and tubular epithelial cells were exposed to 1 200 mg/L uric acid for 24 h. Compared with the control group, 1 200 mg/L uric acid decreased the cell survival rates significantly (P＜0.01, P＜0.01, P＜0.01). Compared with the model group, pretreatment with 0.1, 1, 10, 100 μmol/L iptakalim could remarkably alleviate cellular damages of glomerular endothelium, mesangium cells induced by uric acid (P＜0.05, P＜0.01, P＜0.01, P＜0.01). The K_ATP channel blocker could clearly reduce survival rates of the renal glomerular endothelial, mesangial cells(P＜0.01) and markedly reverse the inhibitory effects of iptakalim on cell death (P＜0.05, P＜0.01), no obvious difference in comparison with the model group (P＞0.05). Compared with the model group, pretreatment with 10, 100 μmol/L iptakalim could notably attenuate cellular damages of tubular epithelial cells induced by uric acid (P＜0.05, P＜0.05). The K_ATP channel blocker could obviously damage the tubular epithelial cells (P＜0.01), no obvious difference in comparison with the model group (P＞0.05). Compared with control group, exposure of renal tubular epithelial, mesangial and glomerular endothelial cells to 1 200 mg/L uric acid for 24 h caused a significant increase in the protein expressions of Kir6.1 and SUR2B(P＜0.05). Compared with the model group, the overexpressions of Kir6.1 and SUR2B were suppressed in presence of iptakalim at a concentration of 10 μmol/L (P＜0.05). These decreases in the expressions of Kir6.1 and SUR2B were prevented by the K_ATP channel blocker, no obvious difference in comparison with the model group (P＞0.05). Compare with the control group, monocytic adhesion to renal glomerular endothelial cells was notably promoted by 1 200 mg/L uric acid for 24 h (P＜0.01). Pretreatment with 10 μmol/L iptakalim for 24 h significantly reduced the monocytic adhesion in comparison with the model group (P＜0.05). It was showed that the inhibitory effects of iptakalim were antagonized by the K_ATP channel blocker, no obvious difference in comparison with the model group (P＞0.05). After stimulating glomerular endothelial cells with 1 200 mg/L uric acid for 24 hours, the secretion of MCP-1 was significantly increased compared to the control group (P＜0.05). Compare with the model group, preincubation with 10 μmol/L iptakalim significantly decreased MCP-1 production (P＜0.05). K_ATP channel blocker suppressed the downregulation of MCP-1 protein synthesis induced by iptakalim. After stimulation with uric acid, translocation of NF-κB from cytoplasms to nuclei of renal glomerular endothelial cells were observed, while that of NF-κB was suppressed in presence of iptakalim at the concentration of 10 μmol/L. This inhibition of NF-κB translocation was clearly prevented by K_ATP channel blocker. Conclusion: These results suggests that a new SUR2B/Kir6.1-type K_ATP channel opener iptakalim plays interventional roles in renal cells damages caused by uirc acid and its mechanism is involved in activating K_ATPchannels .

Objective: To investigate the effects and its mechanisms of silence information regulator 7(SIRT7)on mouse renal podocytes proliferation and apoptosis under high glucose environment. Methods: Mouse renal podocytes cultured with high glucose and treated with different methods were divided into the following groups:control group(Control),high glucose group(HG),high glucose+transfecting with SIRT7 overexpression vetor(pcDNA3.1-SIRT7) group(SIRT7 OE+HG),high glucose+transfecting with the negative control vetor(pcDNA3.1)group(SIRT7 OE-NC+HG),high glucose+transfecting with small interfering RNA-SIRT7 (siRNA-SIRT7) group (siRNA-SIRT7+HG), high glucose+ transfecting with siRNA-SIRT7 control group (siRNA-SIRT7-NC+ HG). Viability of proliferation was examined by CCK-8 method.Rate of apoptosis was detected by flow cytometry. The level of SIRT7 mRNA expression was measured by qRT-PCR. Western blot was performed to detect the protein expression of Nephrin and key factors of Wnt/β-catenin signaling pathway. Results: The CCK-8 result showed that,compared with control group, the proliferative activity of mouse renal podocytes in HG group was decreased (P＜0.05). After transfected with SIRT7 overexpression vetor or small interfering RNA-SIRT7,compared to HG group,the cell proliferation activity was further decreased in siRNA-SIRT7 group(P＜0.05),but it was enhanced in SIRT7 OE+HG group (P＜0.05). The results of flow cytometry showed that compared with the control group, the apoptosis rate of cells in the HG group was increased (P＜0.05). Compared with the HG group, the apoptosis rate of cells in the siRNA SIRT7+HG group was increased significantly(P＜0.05), while that in the SIRT7 OE+HG group was decreased (P＜0.05). Compared with control group,the expressions of Nephrin, Wnt5a and β-catenin were inhibited in HG group (P＜0.05). compared to HG group,siRNA-SIRT7 could down-regulate the expression levels of Nephrin, Wnt5a and β-catenin in siRNA-SIRT7 group (P＜0.05), SIRT7 overexpression could up-regulate the expression levels of Nephrin, Wnt5a and β-catenin in SIRT7 OE+HG group (P＜0.05). Conclusion: The findings suggest that high glucose environment is an important factor to inhibit the proliferation and induce apoptosis of mouse renal podocytes.Overexpression of SIRT7 can reverse the effects by activating Wnt/β-catenin signaling pathway and up-regulating β-catenin expression.

Objective: To investigate the protective effects of erythropoietin derived peptide, also known as spiral B surface peptide (HBSP), on kidney and aggregated proteins (Agrin) levels in acute skeletal muscle strain rats. Methods: Forty SPF grade SD male rats were randomly divided into control group, injury group, HBSP group and EPO group, with 10 rats in each group. Acute skeletal muscle strain animal models were established except the control group. After successful modeling, the rats in HBSP group and EPO group were intraperitoneally injected with 60 μg/kg HBSP and 5 000 U/kg recombinant human erythropoietin (rhEPO), and the rats in the control group and the injured group were intraperitoneally injected with 0.9% normal saline. Renal function was monitored with relevant kits; Hematoxylin-eosin staining was used to observe the pathological morphology of kidney tissue and skeletal muscle strain tissue. The apoptosis rate of renal tissue cells was detected by in situ terminal transferase labeling (TUNEL). Western blot and quantitative polymerase chain reaction (Q-PCR) were used to determine the expressions of Agrin and muscular-specific kinase (MuSK) in the injured skeletal muscle of rats in each group. Results: Compared with the control group, the renal function indexes serum creatinine (Cr), urea nitrogen (BUN) and 24 h urinary protein (UP24) levels of rats in injured group were increased (P＜ 0.05), but the levels of BUN, Cr and UP24 of rats in HBSP group were decreased (P＜0.05). Compared with HBSP group, there were no significant differences in the above indexes in EPO group (P＞0.05). In the control group, the muscle fiber structure was intact, the shape and structure of the fiber bundles were normal, and there was no infiltration of red blood cells and inflammatory cells in the interstitium, and no fibrohyperplasia. In the injured group, the muscle tissue showed sparse and irregular arrangement, and the interstitial widened with a large number of inflammatory cells and red blood cell infiltration. Erythrocytes and inflammatory cells were reduced in HBSP group and EPO group, and the transverse and longitudinal lines of muscle were clear. The glomerular structure of the rats in the fibrohyperplasia control group was intact and no lesions were observed. In the injured group, glomerular hypertrophy and significant matrix hyperplasia were observed, as well as the expansion of renal cysts with vacuolar and significant inflammatory infiltration were observed, and the inflammatory infiltration was reduced in the HBSP and EPO groups. Glomerular hypertrophy and hyperplasia were alleviated. The apoptosis rates of kidney cells in control group, injured group, HBSP group and EPO group were (4.05±0.51) %, (26.30±2.05) %, (14.28±1.62) % and (16.03±1.77) %, respectively, and there were significant differences among these groups (P＜0.05). Compared with control group, the levels of Agrin and MuSK in skeletal muscle pulled tissue were significantly decreased (P＜0.05), and those of HBSP group and EPO group were significantly increased compared with injured group (P＜0.05), but there was no significant difference between HBSP group and EPO group (P＞0.05). Conclusion: Erythropoietin derived peptide (HBSP) has obvious intervention effects on renal function injury in rats with acute skeletal muscle strain, and its mechanisms may be related to reducing the apoptosis rate of renal tissue cells and activating Agrin and MuSK expression.

Objective: To investigate the effect of SIX2 gene on the proliferation of bovine skeletal muscle satellite cells. Methods: Bovine skeletal muscle satellite cells were used as experimental materials, and the expression of SIX2 gene in bovine skeletal muscle satellite cells was detected by real-time quantitative PCR at 24 h, 48 h, and 72 h of proliferation. The SIX2 gene overexpression vector was constructed by homologous recombination. The SIX2 gene overexpression plasmid and the control empty plasmid were transfected into bovine skeletal muscle satellite cells, and each group had three complex Wells. The cell viability was detected by MTT assay at 24 h, 48 h and 72 h after transfection. At 48 h after transfection, the cell cycle was detected by flow cytometry, and the expressions of cell proliferation marker genes were detected by real-time quantitative PCR (qRT-PCR) and Western blot. Results: With the proliferation of bovine skeletal muscle satellite cells, the expression of SIX2 mRNA was increased. Compared with the control group, the expressions of SIX2 mRNA and protein in the SIX2 gene overexpression plasmid group were increased by 18 and 2.6 times, respectively (P＜0.01). The cell viability of the SIX2 gene overexpression plasmid group was increased (P＜0.01), the proportion of G1 cells was decreased by 24.6%, and the proportion of S phase and G2 phase cells was increased by 20.3% and 4.31%, respectively (P＜0.01). The mRNA and protein expressions of Pax7 gene were increased by 15.84 and 1.22 times, respectively, and the mRNA and protein expressions of proliferation marker genes PCNA and CCNB1 were increased by 4.82, 2.23,1.55 and 1.46 times, respectively (P＜0.01). Conclusion: Overexpression of SIX2 gene promotes the proliferation of bovine skeletal muscle satellite cells.

Objective: To investigate the effects of cigarette smoke extract (CSE) on the mitochondrial function of macrophages. Methods: RAW264.7 macrophages were used for the experiment in this study. When the cell density was about 70%, the old culture medium was abandoned, and the 100% CSE stock solution was diluted with serum-free DMEM and FBS into 1%, 5%, 15%, 25% and 90% CSE and added to the well plate. The cell activity of RAW264.7 cells treated with CSE at different concentrations for 24 h was detected by CCK-8 method. Then the optimal CSE concentration was selected to treat cells for 0 h, 24 h, 48 h or 72 h respectively, and CCK-8 assay was used to detect the cell activity of CSE treated cells at different time groups. After the cells were treated with 0%, 5% and 25% CSE for 24 hours, cell necrosis and apoptosis was detected by Annexin V-FITC /PI staining; Mitochondrial membrane damage of RAW 264.7 was detected by mitochondrial membrane potential assay kit with JC-1; Macrophages were stained with ROS-specific dye DCFH-DA, and then Flow cytometer was used to determine the fluorescence and the proportion of ROS-positive macrophages; the enhanced ATP assay kit was used to detect the intracellular ATP concentration. Results: ①Compared with 0% CSE, cell viability was increased significantly in 1% CSE group (P＜0.01), cell viability was decreased significantly when CSE concentration was above 5% (P＜0.05); Macrophages were treated with 5% CSE, and cell viability was decreased significantly with the increase of treatment time (P＜0.01). ②Compared with 0% CSE, 5% CSE and 25% CSE mainly caused macrophage necrosis, decreased mitochondrial membrane potential, increased ROS production and decreased ATP significantly (P＜0.05 or P＜0.01), and the changes were more significant in 25% CSE treatment group(P＜0.05 or P＜0.01). Conclusion: CSE may affect mitochondrial function of macrophages, leading to decreased cell viability and necrosis.

Objective: To investigate the effects of oil-mist particulate matter (OMPM) on cardiac tissue structure fibrosis in rats and the role of epithelial-mesenchymal transition (EMT). Methods: Six-week-old Wistar rats (half male and half female) were randomly divided into 3 groups: control group (without OMPM exposure), low-dose exposure group (50 mg/m³) and high-dose exposure group (100 mg/m³), 18 rats in each group, with 6.5 hours per day of dynamic inhalation exposure. After 42 days of continuous exposure, cardiac tissues were collected for morphological observation; Western blot was used to detect fibrosis markers collagen I and collagen III levels, epithelial marker E-cadherin levels, interstitial markers N-cadherin, fibronectin, vimentin, alpha-smooth muscle actin (α-SMA) levels, and EMT transcription factor Twist protein levels; Real-time polymerase chain reaction (RT-qPCR) was used to detect collagen I and collagen III mRNA levels. Results: After OMPM exposure, myocardial cell edema and collagen fiber deposition were increased gradually with increasing exposure dose. Western blot results showed that compared with the control group, the expression levels of collagen I, collagen III, N-Cadherin, fibronectin, vimentin, α-SMA, and Twist protein were increased significantly in the low-dose exposure group and the high-dose exposure group (P＜0.01), and protein expression levels were higher in the high-dose exposure group than those in the low-dose exposure group (P＜0.01). In contrast, E-Cadherin protein expression levels were decreased significantly, and lower in the high-dose exposure group (P＜0.01). RT-qPCR results showed that compared with the control group, collagen I and collagen III mRNA levels were increased significantly in the low-dose exposure group and the high-dose exposure group (P＜0.01), and were increased with increasing exposure dose. (P＜0.01). Conclusion: OMPM may induce cardiac fibrosis in rats by promoting EMT process.

Objective: To study the effects of estradiol (E2) on alleviating myocardial ischemia/reperfusion(I/R) injury through estrogen receptorβ(ERβ) mediated extracellular regulated protein kinases(ERK) pathway activation. Methods: Eighty-four adult female SD rats were ovariectomized and randomly divided into control group, NC siRNA adeno-associated virus (AAV) group received sham operation, the myocardial I/R injury model was prepared by ligation of the left anterior descending coronary artery in I/R group, E2+I/R group, NC siRNA AAV+I/R group, NC siRNA AAV+E2+I/R group and ERβ-siRNA AAV+E2+I/R group. E2+I/R group, NC siRNA AAV+E2+I/R group and ERβ-siRNA AAV+E2+I/R group were treated with E2 0.8 mg/kg by gavage for 60 days before modeling. NC siRNA AAV+I/R group, NC siRNA AAV+E2+I/R group, and ERβ-siRNA AAV+E2+I/R group were treated with AAV by caudal vein injection 24 h before modeling. After 120 min of reperfusion, the contents of serum lactate dehydrogenase (LDH), phosphocreatine kinase (CK), phosphocreatine kinase isoenzyme (CK-MB), myocardial infarction area and the expressions of ERβ, p-ERK, the contents of tumor necrosis factor-α(TNF-α), interleukin-1β(IL-1 β), malondialdehyde (MDA) and total antioxidant capacity (T-AOC) in myocardium were measured. Results: The contents of serum LDH, CK, CK-MB, myocardial infarction area and the contents of TNF-α, IL-1 β, MDA in myocardium of I/R group were higher than those of the control group, the expression levels of ERβ and p-ERK and the content of T-AOC were lower than those in the control group (P＜0.05). The contents of serum LDH, CK and CK-MB, myocardial infarction area and the contents of TNF-α, IL-1 β and MDA in myocardium of E2+I/R group were lower than those of the I/R group, the expression levels of ERβ and p-ERK and the content of T-AOC were higher than those of the I/R group(P＜0.05). After knockdown ERβ by caudal vein injection of ERβ-siRNA AAV, the contents of serum LDH, CK and CK-MB, myocardial infarction area and the contents of TNF-α, IL-1 β and MDA in myocardium of ERβ-siRNA AAV+E2+I/R group were higher than those of NC-siRNA AAV+E2+I/R, the expression levels of ERβ and p-ERK and the content of T-AOC were lower than those of NC-siRNA AAV+E2+I/R(P＜0.05). Conclusion: E2 has protective effects on myocardial I / R injury in ovariectomized rats, which are related to the promotion of ERβ mediating the activation of ERK pathway, reducing inflammatory and oxidative stress responses.

Objective: To study the protective effects of resveratrol (RSV) on cardiac function in rats with high altitude hypobaric hypoxia and its mechanisms. Methods: Thirty-six rats were randomly divided into control group, hypobaric hypoxia group (HH) and hypobaric hypoxia + RSV group (HH+RSV) according to the random number, 12 rats in each group. Rats in the HH and HH+RSV groups were subjected to chronic long-term high altitude hypobaric hypoxia intervention for 8 weeks in a hypobaric chamber at a simulated altitude of 6 000 m for 20 h / d. The rats of HH + RSV were fed with RSV at a dose of 400 mg/(kg·d). The rats were tested once a week for body weight and twice a week for food intake. Before execution, the rats were tested by blood cell analyzer for routine blood parameters and echocardiogram for cardiac function parameters in each group. The routine blood indexes of each group were measured by blood cell analyzer, the cardiac function indexes of each group were measured by echocardiography, myocardial hypertrophy was evaluated by HE staining, myocardial tissue reactive oxygen levels were evaluated by dihydroethidium (DHE) staining. Oxidative stress was evaluated by serum and myocardial tissue total antioxidant capacity (T-AOC), superoxide dismutase activity (SOD) and malondialdehyde (MDA) content. Results: Compared with the C group, the body mass and food intake of rats were decreased significantly (P＜0.05) in HH group, while compared with the C group, RSV had no significant effects on the body mass and food intake of rats in the HH+RSV group (P＞0.05). Compared with the C group, the levels of erythrocytes and hemoglobin of rats in the HH group were increased significantly (P＜0.05), while the platelet concentration was decreased significantly(P＜0.05); compared with the HH group, the erythrocyte and hemoglobin levels were decreased significantly (P＜0.05) and platelet concentration was increased significantly(P＜0.05) in rats of the HH+RSV group. Compared with the C group, the cardiac coefficient, myocardial fiber diameter and thickness were significantly increased in the HH group (P＜0.05); compared with the HH group, the cardiac coefficient and myocardial fiber thickness were significantly decreased in the HH+RSV group (P＜0.05). Echocardiographic analysis showed a significant increase in ventricular wall thickness (P＜0.05) and a significant decrease in ejection fraction and cardiac output (P＜0.05) in the HH group compared with the C group, and a significant decrease in ventricular wall thickness and a significant improvement in cardiac function (P＜0.05) in the HH+RSV group compared with the HH group. The results of DHE staining showed that myocardial tissue reactive oxygen levels were increased significantly in the HH group compared with the C group (P＜0.05); myocardial tissue reactive oxygen levels were significantly restored in the HH+RSV group compared with the HH group (P＜0.05). The oxidative/antioxidant results showed that the serum and myocardial T-AOC and SOD activities were decreased significantly (P＜0.05) and the MDA level was increased significantly (P＜0.05) in the HH group compared with the C group; the serum and myocardial T-AOC and SOD activities were increased significantly (P＜0.05) and the MDA level was decreased significantly(P＜0.05) in the HH+RSV group compared with the HH group. Conclusion: Long-term plateau hypobaric hypoxia exposure leads to myocardial hypertrophy and reduced cardiac function in rats. Resveratrol intervention significantly improves myocardial hypertrophy and cardiac function in rats caused by altitude hypobaric hypoxia exposure, which is closely related to reducing of reactive oxygen species and improving myocardial oxidative stress levels.

Objective: To investigate the effects of panax notoginseng saponins (PNS) on pulmonary vascular remodeling and SIRT1/FOXO3a/p27 pathway in pulmonary arterial hypertension (PAH) rats. Methods: Male SD rats weighing 200~250g were randomly divided into control group, monocrotaline group (MCT) and monocrotaline + panax notoginseng saponins group (MCT+PNS), with 10 rats in each group. The rats in control group were injected intraperitoneally with normal saline 3 ml/kg on the first day, then injected intraperitoneally with normal saline 2.5 ml/kg every day. The rats in MCT group were injected intraperitoneally with MCT 60 mg/kg on the first day, followed by daily injection of normal saline 2.5 ml/kg. In MCT+PNS group, 60 mg/kg MCT was injected intraperitoneally on the first day, and 50 mg/kg PNS was injected intraperitoneally every day. The above models were fed conventionally for 4 weeks. After the modeling was completed, the mean pulmonary artery pressure (mPAP) and right ventricular systolic pressure (RVSP) of rats in each group were detected by right heart catheter method, weighed and calculated right ventricular hypertrophy index (RVHI), and the pulmonary vascular structure and morphological changes were observed by HE and Masson staining. The protein and gene expressions of SIRT1, FOXO3a, p27, PCNA and Caspase-3 were detected by qPCR and Western blot. Results: Compared with control group, mPAP, RVSP and RVHI in MCT group were increased significantly (P＜0.01), pulmonary vessels were thickened significantly and collagen fibers were increased, protein and gene expressions of SIRT1, FOXO3a, p27 and Caspase-3 were decreased (P＜0.05 or P＜0.01). The protein and gene expressions of PCNA were increased (P＜0.05). Compared with MCT group, the levels of mPAP, RVSP and RVHI in MCT+PNS group were decreased significantly (P＜0.05 or P＜0.01), pulmonary vascular thickening was alleviated and collagen fibers were reduced. The protein and gene expressions of SIRT1, FOXO3a, p27 and Caspase-3 were increased (P＜0.05 or P＜0.01), while the protein and gene expressions of PCNA were decreased (P＜0.05 or P＜0.01). Conclusion: Panax notoginseng saponins can relieve pulmonary vascular remodeling in rats with pulmonary hypertension by activating SIRT1/FOXO3a/p27 pathway.

Objective: To investigate the protective effects of salidroside on endothelial cells in rats with frostbite after chronic hypoxia. Methods: Healthy male SD rats, randomly divided into 3 groups with 10 rats in each group, which included the sham injury group, the model group, and the model +salidroside group. The rats in each group were placed in a composite low-pressure chamber to simulate a environment with a pressure of 54.1 kpa and a temperature of 23~25°C. The rats were exposed to hypoxia under these conditions for 14 days, during the experimental time the rats in the model+salidroside group were treated with 50 mg/kg salidroside daily. After the rats were removed from the low-pressure chamber, except for the sham injury group, frozen iron sheets were applied tightly to the back of the rats for 30 s, supplemented with low temperature for frostbite modeling. Blood and skin tissues were collected at 12 hours after modeling for testing. The structural changes in tissue and vascular endothelial cells were observed in the frostbite region. Vascular endothelial cell particulate EMP levels were detected. The levels of ICAM-1, sEPCR, vWF, ET-1 and NO secretion were determined. The expression levels of HIF-1α, p-PI3K, p-Akt and VEGF were detected by Western blot. Results: Salidroside could effectively reduce skin collapse in frostbitten areas. It could reduce the injury of frostbitten tissues, and improve the subcutaneous tissue necrosis and inflammatory cell infiltration. The autophagy of vascular endothelial cells was reduced. Compared with the model group (0.250±0.165)%, the expression of EMPs in the model+salidroside group (2.453±0.196)% was increased significantly (P＜0.01). In addition, the contents of NO (2.622±0.219)pg/ml was also significantly higher than that of the model group (1.616±0.152)pg/ml (P＜0.01), and the content of vWF (233.50±13.43)pg/ml was lower than that of the model group (315.60±8.78)pg/ml (P＜0.05). There was no significant difference in the levels of ICAM-1, sEPCR and ET-1. Salidroside significantly decreased the expressions of p-PI3K, p-Akt, VEGF and HIF-1α protein in vascular endothelial cells of rats with frostbite (P＜0.01). Conclusion: Salidroside can reduce endothelial cell damage, reduce endothelial cell autophagy and promote endothelial cell regeneration. Based on the PI3K/Akt pathway, salidroside has a good protective effect on endothelial cells of rats with frostbite after chronic hypoxia.

Objective: To investigate the effects of Rosa roxburghii on insulin resistance in obese rats and the regulation of phosphatidylinositol 3-kinase (PI3K)/ protein kinase Bβ(PKB_β/Akt2)/ glucose transporter 4(GLUT4) signaling pathway. Methods: Five-week-old male SD rats were randomly divided into normal control group (NC), model group (M), positive control group (PC), low-dose rosa roxburghii group (LD) and high-dose rosa roxburghii group (HD), with 10 rats in each group. The rats in the NC group were fed with normal diet, while those in the M, PC, LD and HD groups were fed with high-fat diet. From the 13th week, according to the dose standard of 6 ml/kg, rats in the LD group were intragastrically administered with 100 mg/kg Rosa roxburghii Tratt, the HD group were treated with 300 mg/kg Rosa roxburghii Tratt, the PC group were treated with 0.11 g/kg Chiglitazar sodium, and the NC and M groups were intragastrically administered with the same volume of normal saline. The body weight was measured every week until 20 weeks. The rats were sacrificed 24 h after the last experiment. Blood and skeletal muscle were collected. Serum total cholesterol (TC) and triglyceride (TG) contents were detected by colorimetric method, serum superoxide dismutase (SOD) activity was detected by xanthine oxidase method, serum malondialdehyde (MDA) content was detected by thiobarbituric acid method, blood glucose (FBG) value was detected by glucose oxidase method, insulin (FINS) content was detected by ELISA, and PI3K, Akt2, and GLUT4 protein and gene expressions were detected by Western blot and reverse transcription-polymerase chain reaction (RT-PCR). Results: Compared with the NC group, the body weight, serum MDA, TG, TC, FBG, FINS, HOMA-IR levels in the M group were significantly increased (P＜0.01), while SOD activity, PI3K、Akt2、GLUT4 protein and mRNA expression levels were significantly increased(P＜ 0.01). Compared with group M, the body weight, serum MDA, TG, TC, FBG, FINS, and HOMA-IR were decreased significantly in LD group, HD group and PC group (P＜0.05 or P＜0.01), while SOD activity, PI3K, Akt2, GLUT4 protein and mRNA expression levels were increased significantly (P＜0.05 or P＜0.01). Conclusion: Rosa roxburghii can improve insulin resistance in obese rats by antioxidant stress and up-regulating the expressions of PI3K, Akt2, and GLUT4 proteins and genes, which may be related to the PI3K/Akt2/GLUT4 signaling pathway.

Objective: To investigate the effects of aerobic intermittent exercise on the expressions of KLF15/mTOR related proteins to improve skeletal muscle lesions in type 2 diabetes rats. Methods: The experimental model of type 2 diabetes rats was established by feeding high-fat diet for 4 weeks and intraperitoneal injection of streptozotocin (STZ). After modeling, rats were randomly divided into two groups: diabetes model group (DM), diabetes+exercise group (DE), and normal rats were set as control group (C), 10 rats in each group. Group DE was given 8-week aerobic intermittent treadmill exercise intervention, while group C was not given any intervention. At the end of the experiment, the expressions of KLF15, mTOR, p-mTOR, and cleared caspase-3 in gastrocnemius muscle were detected by Western blot. The histopathologic changes of gastrocnemius were observed under microscope; skeletal muscle cells apoptosis rates and muscle mass were examined respectively using HE staining and TUNEL fluorescence staining. At the same time, changes of blood glucose and serum insulin, and weight were examined in the end of the experiment. Results: ①Compared with group C, the wet weight of gastrocnemius muscle and body weight, ratio of wet gastrocnemius muscle and body weight in group DM were decreased(P＜0.05 or P＜0.01); compared with group DM, the wet weight of gastrocnemius muscle, ratio of wet gastrocnemius muscle and body weight in the group DE were increased significantly (P＜0.05). ②Compared with group C, the fasting blood glucose level of group DM was increased significantly (P＜0.01), while serum insulin level of the group DM was decreased significantly(P＜0.01);compared with group DM, the above indexes were opposite in the group DE with intervention(P＜0.05). ③Compared with group C, the morphology of skeletal muscle cells in group DM was abnormal, the number of muscle nuclei was increased, the transverse lines were blurred and disappeared, the sarcomere was broken, and some muscle fibers were dissolved. Compared with group DM, the abnormal cell morphology, segmental injury of sarcomere and dissolution of muscle fibers in group DE were improved. The sarcolemma was more complete and the arrangement of muscle nuclei was more orderly. ④Compared with group C, the expressions of KLF15 and cleaved caspase-3, cells apoptosis rates in group DM were increased significantly(P＜0.01), while p-mTOR/mTOR level was decreased(P＜0.01) ; compared with group DM, the above indexes were opposite in the group with intervention(P＜0.05 or P＜0.01). Conclusion: Aerobic intermittent exercise is beneficial to improve the skeletal muscle pathological changes in type 2 diabetes rats, which may be due to the effective regulation of KLF15/mTOR related protein expression and the reduction of apoptosis damage.

Objective: To investigate the effects of adipokines chemerin on the improvement of islet function caused by exercise in mice with diabetes, and the possible mechanism of glucagon-like peptide 1 (GLP-1). Methods: Male ICR mice were randomly divided into a control group fed with normal diet (Con, n=6) and a diabetic modeling group fed with 60% kcal high-fat diet (n=44). After 6 weeks, the diabetic modeling group was once given a fasting intraperitoneal injection of streptozotocin (100 mg/kg). The successfully modeled mice were divided into diabetes group (DM), diabetes plus exercise group (EDM), and diabetes plus exercise and exogenous chemerin group (EDMC), 6 in each group. Mice in exercise groups participated in a six-week modest intensity treadmill running exercise with a gradually increased load. Mice in the EDMC group were intraperitoneally injected with exogenous chemerin (8 μg/kg) from the 4th week of the exercise period, six days per week, and one time per day. And the other groups were untreated. Adipose chemerin knockout mice were constructed. Then they and the control mice were divided into 6 groups (n=4): Normal diet control group (Con-ND), Normal diet chemerin knockout heterozygote mice group (Chemerin(+/-)-ND), Normal diet chemerin knockout homozygotes mice group(Chemerin(-/-)-ND), High-fat diet control group (Con-HFD), High-fat diet chemerin knockout heterozygote mice group (Chemerin(+/-)-HFD), High-fat diet chemerin knockout homozygotes mice group (Chemerin(-/-)-HFD). They were fed with normal or high-fat diet for 11 weeks and oral glucose tolerance test (OGTT) was conducted. After the mice of each group were executed under anesthesia, the samples such as pancreas and colon were collected. Fasting blood glucose (FBG) and fasting insulin (FINS) levels in mice were measured, and the insulin resistance index (HOMA-IR) was calculated. HE staining was used to observe the structure of islets. ELISA was used to detect the GLP-1 level in serum. The mRNA levels of proglucagon (GCG) and chemerin in the colon were measured by real-time PCR. And the protein levels of GCG and chemerin in the colon were detected by Western blot. Results: Compared with the DM group, the vacuolar degeneration and shrinkage of islet cells in the EDM group were reduced, the islet structure was improved, while the levels of FINS, HOMA-IR and FBG were decreased significantly (P＜0.05 or P＜0.01). The colon and serum chemerin levels were decreased significantly(P＜0.05), while the colonic GCG mRNA and protein levels were increased significantly (P＜0.05 or P＜0.01). Compared with the EDM group, the islet cells in the EDMC group were shrunken, with unclear borders. The structure of the islets was damaged, and the levels of FINS, HOMA-IR and FBG were increased significantly (P＜0.01), while the mRNA and protein levels of GCG were decreased significantly (P＜0.05 or P＜0.01). Compared with the Con-HFD group, the blood glucose at 30, 90 and 120 min after oral glucose in the chemerin (-/-)-HFD group was significantly lower (P＜0.01), and the area under the blood glucose time curve was significantly lower (P＜0.01). The islets had clear structure, regular shape and well-defined boundaries, while the serum GLP-1 and colonic GCG protein levels were increased significantly(P＜0.05). Conclusion: Aerobic exercise improves the structure and function of pancreatic islets by reducing the level of chemerin in diabetes mice, which is related to the negative regulation of chemerin on GLP-1 level.

Objective: To investigate the effects of adding whole body vibration (WBV) to routine exercise regimen of pulmonary rehabilitation (PR) on bone strength, lung function and exercise ability of elderly patients with stable chronic obstructive pulmonary disease (COPD) complicated with osteoporosis (OP). Methods: Thirty seven elderly patients with stable COPD were randomly divided into control group (group C, n=12, age: 64.6±3.8 years), conventional PR group (PR group, n=12, age: 66.1±4.9 years), and whole body vibration combined PR group (WP group, n=13, age: 65.5±3.3 years). Before intervention, X-ray and computerized tomography bone scan, bone metabolic markers, pulmonary function, cardiopulmonary exercise, 6-minute walking and isokinetic muscle strength were performed, and then intervened for 36 weeks, three times/week, among which group C subjects were given routine treatment, PR group added aerobic running and static weight resistance on the basis of routine treatment, and WP group added WBV on the basis of PR group intervention. After the intervention, the same indicators were detected. Results: Compared with before the intervention, the pulmonary function indexes of each group were significantly improved after the intervention (P＜0.05), and the bone mineral density and bone microstructure indexes of the patients in the WP group were also significantly improved (P＜0.05). Compared with group C and group PR, the bone mineral density, bone microstructure, parathyroid hormone (PTH), insulin-like growth factor-1 (IGF-1), interleukin-6 (IL-6), osteocalcin (OCN) and other bone metabolism indexes, knee flexion, peak extension torque, fatigue index and muscle strength of patients in WP group were significantly improved (P＜0.05). Conclusion: Adding WBV to the conventional PR regimen can improve the bone strength, lung function and exercise capacity of elderly patients with COPD complicated with OP, and may be able to make up for the deficiency of the current conventional PR regimen for insufficient muscle and bone stimulation.

Objective: To investigate the effects of moderate intensity continuous training (MICT) and high intensity intermittent training (HIIT) on the ultrastructure of myocardium and soleus in rats with high fat diet, and to explore the mechanisms. Methods: 5-week-old male SD rats were randomly divided into normal diet quiet group (C), high-fat diet quiet group (F), high-fat MICT group (M) and high-fat HIIT group (H), with 8 rats in each group, and the fat content of the high-fat dietary feed was 45%. The M and H groups were given 12 weeks of treadmill running with an incline of 25°. The M group was given continuous exercise with 70%VO₂max intensity, and the H group was given intermittent exercise with 5 min 40%~45%VO₂max and 4 min 95%~99%VO₂max intensity successively. After the intervention, the contents of free fatty acid (FFA), triglyceride (TG), high density lipoprotein cholesterol (HDL) and low density lipoprotein cholesterol (LDL) in serum were detected. Transmission electron microscopy was used to observe the ultrastructure of myocardium and soleus in rats. Western blot was used to detect the protein expressions of AMPK, malonyl-CoA decarboxylase (MCD) and carnitine palmitoyl transterase 1 (CPT-1) in myocardium and soleus. Results: Compared with C group, the body weight, Lee's index, the contents of LDL, TG and FFA in serum were increased, the content of HDL was decreased (P＜0.05), the protein expressions of AMPK and CPT-1 in myocardium and soleus were increased, the protein expression of MCD was decreased (P＜0.05), and the ultrastructure was damaged in group F. Compared with F group, the body weight and Lee's index were decreased, the contents of LDL and FFA in serum were decreased (P＜0.01), the protein expressions of AMPK, MCD and CPT-1 in myocardium were increased, and the protein expressions of AMPK and MCD in soleus were increased (P＜0.05), and the ultrastructural damage was attenuated in M and H groups. Compared with M group, the content of HDL in serum was increased (P＜0.01), the protein expressions of AMPK and MCD in myocardium were increased, and the ultrastructural damage was mild, the protein expression of AMPK in soleus was decreased, the protein expression of MCD in soleus was increased (P＜0.05), and the ultrastructural damage was severe in group H. Conclusion: MICT and HIIT have different effects on the ultrastructure of myocardium and soleus in high-fat diet rats by intervening the protein expression of AMPK, MCD and CPT-1.

Objective: To investigate the effects of one-time exhaustive exercise on coagulation state in rats and its mechanism. Methods: Forty-eight SD rats were randomly divided into control group and exhaustive exercise group, 24 rats in each group. Rats in exhaustive exercise group were trained with treadmill training for 25～50 min at a time on non-slope treadmill and the initial speed of 5 m/min was uniformly accelerated to 25 m/min until the rats exhausted. Thrombelastography (TEG) was used to monitor the coagulation function of rats after training. The ligation model of inferior vena cava (IVC) was established to evaluate thrombosis. The phosphatidylserine (PS) exposure and Ca²⁺ concentration were detected by flow cytometry. The production of FXa and thrombin was detected by microplate reader. The clotting time was measured by using coagulometer. Results: Compared with the control group, the blood of rats in the exhaustive exercise group exhibited hypercoagulable state. The probability of thrombus formation, weight, length and ratio in the exhaustive exercise group were significantly higher than those in the control group (P＜0.01). The levels of PS exposure and intracellular Ca²⁺ concentration of red blood cells (RBCs) and platelets in the exhaustive exercise group were increased significantly (P＜0.01). The blood clotting time of RBCs and platelets was shortened (P＜0.01), and the production of FXa and thrombin was increased significantly (P＜0.01) in the exhausted exercise group, and both were inhibited by lactadherin (Lact, P＜0.01). Conclusion: The blood of exhaustive exercise rats is in a hypercoagulable state and the risk of thrombosis is increased. The increased PS exposure of RBCs and platelets caused by exhaustive exercise may be an important mechanism of thrombosis.

Objective: This experiment was designed to observe the effects of aerobic exercise on depressive behavior in rats induced by chronic unpredictable mild stress (CUMS), and to explore the possible mechanism by detecting the proteins related to mitochondrial autophagy. Methods: SD rats were randomly divided into three groups: blank control group (C, n=12), depression model group (D, n=12) and post-depression exercise group (D+E, n=12). Group D and D+E were modeled with CUMS for 28 days, and group D+E underwent aerobic exercise intervention for 4 weeks after model establishment. Then the behavior of rats was evaluated. The concentrations of whole brain dopamine and norepinephrine were determined by ELISA kits. The morphology and structure of mitochondria in the frontal lobe were observed with the transmission electron microscope (TEM). Mitochondrial autophagy lysosomes were localized by immunofluorescence colocalization. The expressions of LC3 and P62 proteins in the frontal lobe were measured with Western Blotting. The relative content of mitochondrial DNA was detected using Real-time PCR. Results: ①Compared with group C, the sucrose preference ratio in group D was decreased significantly(P＜0.01); Compared with group D, the sucrose preference ratio in group D+E was increased significantly (P＜0.01). ②In the open field experiment, compared with group C, group D had a significant decrease in activity, average speed and total distance (P＜0.05); Compared with group D, the average rate of activity in group D+E was significantly higher (P＜0.05). ③ELISA results showed that the levels of whole brain dopamine and norepinephrine were significantly lower in group D rats than those in group C (P＜0.05). ④Under transmission electron microscopy, compared with group C, group D had different degrees of mitochondrial swelling, decreased crest density, and intermembrane space dilation.; Compared with group D, a significant increase in mitochondrial autophagosomes and autophagic lysosomes was observed in neurons in group D+E. Increased co-localization of mitochondria with lysosomes in the D+E group could be observed under fluorescence microscopy. ⑤Compared with group C, the expression of P62 was increased significantly(P＜0.05), and LC3II/LC3I ratio was decreased significantly (P＜0.05) in group D; Compared with group D, LC3II/LC3I ratio was significantly higher in group D+E than that in group D (P＜0.05). ⑥Compared with group C, the relative number of mitochondrial DNA in the frontal lobe of group D was increased significantly (P＜0.05). Conclusion: Aerobic exercise has a significant improvement effect on depression induced by CUMS in rats, and its mechanism may be related to the upregulation of the level of linear autophagy.

Objective: To investigate the effects of bosutinib on the malignant behavior of thyroid papillary carcinoma B-CPAP cells and its possible mechanisms. Methods: Thyroid papillary carcinoma B-CPAP cells were cultured in vitro with a concentration gradient of(1、2、3、4 and 5 μmol/L)bosutinib intervened for 24 hours, DMSO was used as the control group. Five parallel compound holes were set in each group. Cell counting kit (CCK-8 method) method was used to detect cell proliferation. Transwell assay and cell wound healing assay were used to detect cell invasion and migration. TUNEL staining assay and flow cytometry were used to detect cell apoptosis. Western blot was used to detect the expressions of autophagic proteins (Beclin-1, LC3, p62) and signal pathway proteins (SIK2, p-mTOR, mTOR, p-ULK1, ULK1). Results: Compared with the control group, the cell proliferation activity, migration ability and invasion ability were decreased (P＜0.01), while the cell apoptosis rate was increased (P＜0.01) in the bosutinib concentration groups of 2, 3, 4 and 5 μmol/L . In the concentration groups of 4 and 5 μmol/L, the expression of Beclin-1 (P＜0.05), LC3- Ⅱ/LC3- Ⅰ (P＜0.05), SIK2 (P＜0.01) and p-ULK1 (P＜0.01) protein was decreased, while the expression of p62 (P＜ 0.05) and p-mTOR (P＜0.01) protein was increased. Conclusion: Bosutinib may inhibit the autophagy of thyroid papillary carcinoma cells through SIK2-mTOR-ULK1 signaling pathway to inhibit their proliferation, invasion and migration and promote apoptosis, thereby weakening their malignant behavior.

Objective: To investigate the effects of fucoidan inducing impairment of human osteosarcoma cell 143B, as well its mechanisms. Methods: After 143B cells were treated with different concentrations of FUC (0, 0.5, 1, 10, 100, 400, 800 μg/ml) for 48 h, the cell viability and dehydrogenase (LDH) level were detected by MTT assay and chemical colorimetry with six multiple wells for each concentration. Based on MTT results, we determined the value of IC₅₀ was 244.5 μg/ml. The follow-up experiments were divided into control group (without FUC), FUC (10 μg/ml)-treated group, FUC (100 μg/ml)-treated group, FUC (400 μg/ml)-treated group and positive group (resveratrol, 40 μmol/L). There were four multiple wells for each concentration, and each experiment was repeated at least three times. Flow cytometry was performed to detect cell apoptosis and intracellular reactive oxygen species (ROS) level; acridine orange (AO) staining and lyso-tracker red staining were used to observe the autophagolysosome formation; chemical colorimetric analysis was performed to determine malondialdehyde (MDA) content and the activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px); Western blot was used to detect protein expressions of nuclear factor E2-associated factor 2 (Nrf2), heme oxygenase 1 (HO-1) and autophagy-associated proteins including microtubule-associated light chain protein 3 (LC-3), Atg7, Beclin-1 and p62. Results: Compared with control group, the cell viability was decreased significantly in FUC (100～400 μg/ml)-treated groups (P＜0.01); LDH levels in the supernatant (P＜0.05 or P＜0.01), the percentage of cell apoptosis (P＜0.01), intracellular ROS level and MDA content (P＜0.01) were increased remarkably; protein expressions of Atg7 and Beclin-1 were upregulated (P＜0.05 or P＜0.01); the conversion from LC-3I to LC-3II was significant (P＜0.01) together with elevation of autophagolysosome formation (P＜0.05 or P＜0.01); while the activities of SOD and GSH-Px and protein expressions of Nrf2, HO-1 and p62 were decreased remarkably (P＜0.05 or P＜0.01). Conclusion: FUC (100～400 μg/ml) treatment induces oxidative damage and autophagic death in osteosarcoma 143B cells.

Objective: To investigate the effects of ACC1 knockdown on human glioma U251 cell migration and its molecular mechanisms. Methods: Human glioma U251 cell line was used. The experiment was carried out in three steps. Experiment 1: knockdown of ACC1 in U251 cells (shACC1) and its control (NC) U251 cells were established by transfection of shACC1 lentivirus and negative control virus. The cell migration was detected by Transwell migration assay and scratch test. Western blot (WB) was performed to detect the levels of ACC1, Vimentin, Fibronectin, N-cadherin, E-cadherin and Slug proteins. Experiment 2: RT-qPCR and WB were performed to verify the RNA-seq result, upregulation effect of ACC1 knockdown on PAI-1 in U251 cells. The cells then were treated with PAI-1 inhibitor PAI-039, and the cell migration was detected by Transwell migration assay and scratch assay. The protein levels of ACC1, PAI-1, Vimentin, Fibronectin, N-cadherin, E-cadherin and Slug were examined by WB. Experiment 3: the molecular mechanisms of knocking down ACC1 to increase PAI-1 were explored. The cells were treated with acetyltransferase inhibitor C646, and cell migration was examined by Transwell migration assay and scratch assay. WB was conducted to test the levels of ACC1, H3K9ac, PAI-1, Vimentin, Fibronectin, N-cadherin, E-cadherin and Slug proteins. Each experiment was repeated three times. Results: Experiment 1: lentivirus transfection was performed on glioma U251 cells. Compared with NC group, the expression level of ACC1 in shACC1 group was decreased significantly, indicating that lentivirus transfection was successful (P＜0.01), and the number of migrated cells in shACC1 group was increased significantly (P＜0.01). Migration-related proteins Vimentin, Fibronectin, N-cadherin and Slug were up-regulated, while E-cadherin was down-regulated (P＜0.01). Experiment 2: Compared with NC group, PAI-1 mRNA level in shACC1 group was up-regulated. Compared with control group, cell migration in shACC1+PAI-039 group was decreased (P＜0.01), and migration-related proteins Vimentin, Fibronectin, N-cadherin, and Slug were up-regulated. E-cadherin expression was down-regulated (P＜0.01). Experiment 3: Compared with NC group, the concentration of acetyl-coA and the expression level of H3K9ac in shACC1 group were increased significantly (P＜0.01); After further treatment with histone acetyl transferase inhibitor C646, PAI-1 mRNA level was decreased, cell migration number and H3K9ac expression level were decreased in shACC1+C646 group compared with control group (P＜0.01). Migration-related proteins Vimentin, Fibronectin, N-cadherin and Slug were up-regulated, while E-cadherin was down-regulated (P＜0.01). Conclusion: Knockdown of ACC1 promotes the migration of human glioma U251 cells by increasing histone acetylation which elevates the level of PAI-1.

Objective: To investigate the effects of propranolol on the subcutaneous tumorigenesis of esophageal squamous cell carcinoma (ESCC) cells and the proliferation, migration, cell cycle, apoptosis and autophagy of ESCC cells and its possible molecular mechanisms. Methods: The cell proliferation was detected by MTT (methyl thiazol tetrazolium) assay: ESCC Eca109, KYSE-450 and TE-1 cells were routinely cultured. PBS (Phosphate buffer saline) group (without propranolol) and treated groups (40, 60, 80, 100 μmol/L propranolol) were set up with 5 wells in each group. After treatment for 0, 24, 48, 72 h, 10 μl (5 mg/ml) of MTT was added to each well, and the absorbance was measured at 490 nm. The cell migration was tested by Transwell assay: ESCC Eca109, KYSE-450 and TE-1 cells were routinely cultured, and PBS group (without propranolol) and treated groups (40, 60 μmol/L) were set up with 2 wells in each group. Photos were taken 40 h later, and the experiment was repeated for three times before statistical analysis. The cell cycle and apoptosis were detected by flow cytometry assay: ESCC Eca109, KYSE-450 and TE-1 cells were routinely cultured. PBS group (without propranolol) and treated group (80 μmol/L) were set up, fixed, stained, and fluorescence at 488 nm was detected. The protein levels were detected by Western blot: ESCC Eca109 and KYSE-450 cells were routinely cultured. PBS group (without propranolol) and treated groups (60, 80 μmol/L) were set up followed by gel electrophoresis, wet membrane transfer, and ECL imaging. The experiment was repeated for three times and then analyzed statistically. Subcutaneous tumor formation experiment in nude mice: 10 nude mice were assigned PBS group (without propranolol) and treated group (with propranolol). Five mice in each group were inoculated with 5×10⁶ cells/100 μl (Eca109) into the right underarm. The treated group was given a gavage of 0.4 ml/kg (6 mg/kg) every other day, and the tumor size was measured every other day for 3 weeks. After 20 days, the nude mice were dislocated and sacrificed to take tumor tissue. Result: The results showed that propranolol inhibited the proliferation of Eca109, KYSE-450 and TE-1 cells with IC₅₀ of around 70 μmol/L for 48 h. Eca109, KYSE-450 and TE-1 cell migration was inhibited by propranolol in a dose-dependent manner (P＜0.05); Propranolol blocked the cell cycle of Eca109 in G2/M phase, blocked the cell cycle of KYSE-450 and TE-1 in G0/G1 phase, and promoted apoptosis of three kinds of cells (P＜0.05). The results of cell fluorescence showed that LC3 fluorescence intensity of TE-1 was increased after 12 h, 24 h and 36 h treatment with propranolol (P＜0.05). Western blot results showed that compared with PBS group, the protein expressions of p-mTOR, p-Akt and cyclin D1 were down-regulated, while cleaved caspase 9 level was up-regulated (P＜0.05). The results of subcutaneous tumor formation in nude mice showed that the tumor weight of PBS group was (0.91±0.05)g, and that of the experimental group was(0.65±0.12)g, the difference was statistically significant (P＜0.05). Conclusion: Propranolol inhibits the proliferation, migration and cell cycle,promotes apoptosis and autophagy of ESCC cells, and inhibits subcutaneous tumor growth in nude mice. The mechanism might be related to the inhibition of PI3K/AKT/mTOR signaling pathway.

Objective: To investigate the effects of polystyrene nanoplastics (PS-NPs) exposure during gestation on the growth and neurotoxicity of fetal rats. Methods: Twenty-seven SD pregnant rats were randomly divided into 9 groups with three rats in each group. The experimental group of PS-NPs was given 0.5, 2.5, 10 and 50 mg/kg of PS-NPs suspension with different particle sizes (25 and 50 nm) by gavage, wihe the control group was given ultrapure water by gavage. The time of gavage is from the 1st to the 18th day of pregnancy. The morphological changes of the placenta were observed; compare the number of male and female fetuses, live/dead/absorbed fetuses, body weight, body length, placental weight, and organ coefficients of kidney, liver, brain and intestine of fetal rats; the prefrontal cortex, hippocampus and striatum of the fetal rats were taken to measure related biochemical indicators. Results: Compared with the control group, the placenta of the PS-NPs exposed group was found to have structural damage, which increased in a dose-dependent manner. The area ratio of trophoblast was significantly increased (P＜0.05), and the area ratio of labyrinth was significantly decreased (P＜0.05); In the prefrontal cortex, hippocampus and striatum of fetal rats, the levels of IL-1β, IL -6 and TNF-α were significantly increased in the 10 and 50 mg/kg PS-NPs exposed group (P＜0.05), and more significantly elevated in the 25 nm group than those in the 50 nm group at 10 mg/kg exposure (P＜0.05) the CAT activity was significantly decreased in 2.5, 10 and 50 mg/kg PS-NPs exposure groups (P＜0.05), while the SOD and GSH-Px activities were significantly decreased in 25 nm exposure groups and 2.5, 10 and 50 mg/kg 50 nm PS-NPs exposure groups (P＜0.05), the MDA content was significantly increased in 10, 50 mg/kg 25 nm PS-NPs exposure groups and 50 mg/kg 50 nm PS-NPs exposure groups (P＜0.05). Conclusion: Maternal PS-NPs exposure during gestation may affect the growth and development of fetal rats by damaging the placental barrier and produce neurotoxicity in fetal rats, causing oxidative stress and inflammatory responses in various brain regions, and smaller particle sizes and higher doses of polystyrene nanoplastic exposure have more significant neurotoxic effects on the offspring.

Objective: To study the effects of Cistanche deserticola and its active components Cistanche deserticola polysaccharide and Echinacoside on intestinal flora of antibiotic-associated diarrhea (AAD) mice. Methods: Forty-eight Balb/c mice were randomly divided into control (Con) group, AAD Group, inulin (Inu) group, Cistanche deserticola (RCR) group, Cistanche deserticola polysaccharide (RCRDT) group and Echinacoside (Ech) group with 8 mice in each group. The diarrhea model of mice was induced by intragastric administration of lincomycin hydrochloride(3 g/kg) for 7 days, and then treated by intragastric administration of INU(5 g/kg), RCR(5 g/kg), RCRDT(200 mg/kg) and ECH (60 mg/kg),0.2 ml once a day for 7 days, Con group and AAD group were given the same volume of normal saline. By observing general signs of mice, colon HE staining, 16S rDNA high-throughput sequencing analysis, the effects of Cistanche deserticola, Cistanche deserticola polysaccharide and Echinacea glycoside on the imbalance of intestinal flora induced by antibiotics in mice were evaluated. Results: Compared with Con group, AAD group mice lost weight, presented obvious diarrhea symptoms, inflammatory changes in colon tissue and decreased intestinal flora diversity (P＜0.05) indicating the success of the model. Compared with AAD group, the weight and diarrhea of INU, RCR, RCRDT and ECH groups were significantly improved, and the colon pathology of ECH group was restored to normal level. Compared with AAD group, RCR group, RCRDT group and ECH group had significantly decreased intestinal Firmicutes, increased Blautia and Lachnoclostridium, and decreased Clostridium_sensu_stricto_1 (P＜0.05) . In ECH group, the abundance and diversity of intestinal microflora were returned to normal level, and the structure of intestinal microflora was well adjusted, the contents of Bacteroides, Flavonifractor, Agathobacter, Lachnoclostridium and Prevotella-9 were increased (P＜0.01). Conclusion: Both Cistanche deserticola and its active components cistanche deserticola polysaccharide and echinacoside can regulate the intestinal flora imbalance caused by antibiotics and improve the symptoms of AAD, especially echinacoside.

Objective: To investigate the effects of Mijian Daotong Bowel Suppository (MJDs) on the compound diphenoxylate induced constipation model of male rats and its mechanisms. Methods: Sixty SD male rats were randomly divided into blank group, model group, positive group and MJDs group. The constipation model was established by using compound diphenoxylate gavage. The rats in blank group and model group were treated with saline by enema, the rats in positive group and MJDs group were given Kaisailu and honey decoction laxative suppository by enema, respectively, once a day for 10 days. The body weight, fecal water content, gastric emptying rate (GER) and carbon ink propulsion rate (CIPR) of rats were observed during modeling and administration. The effects of MJDs on the pathological changes of colon tissue in constipation rats were investigated by hematoxylin-eosin (HE) staining. The effect of MJDs on 5-hydroxytryptamine (5-HT) in the colon of constipation rats was investigated by ELISA kit. The effects of MJDs on the expressions of aquaporins 3 (AQP3) and aquaporins 4 (AQP4) in the colon of constipation rats were detected by immunohistochemistry. Results: After 10 days of administration, compared with the blank group, the body weight, fecal water content, carbon ink propulsion rate and colon 5-HT content in the model group were decreased significantly, while the expression levels of AQP3 and AQP4 in the colon were increased significantly (P＜0.05, P＜0.01). Compared with the model group, the fecal water content and colon 5-HT content in the positive group were increased significantly, and the expressions of AQP3 and AQP4 in the colon were decreased significantly. The body weight, fecal water content and colon 5-HT content in the MJDs group were increased significantly, and the expressions of AQP3 and AQP4 was decreased significantly (P＜0.05, P＜0.01). Compared with the positive group, the fecal water content of the MJDs group was decreased significantly, and the expressions of AQP3 and AQP4 in the colon of the MJDs group was decreased significantly (P＜0.05, P＜0.01). Gastric emptying rate was not statistically significant difference between the groups. Conclusion: MJDs has good therapeutic effects on constipation, and its mechanisms may be related to up-regulating the content of 5-HT in the colon and down-regulating the expressions of AQP3 and AQP4 in the colon.

Objective: To investigate the intervention effects of curcumin (Curc) on liver injury induced by chronic alcohol addiction in mice. Methods: Thirty Balb/c mice were randomly divided into normal control group (Control), model group (Model), low-dose Curc group (5 mg/kg, Curc-L), medium dose Curc group (10 mg/kg, Curc-M) and high-dose Curc group (15 mg/kg, Curc-H), with 6 mice in each group. The chronic alcohol addiction liver injury model was prepared with 20% liquor. The mice in control group were given 2 ml of normal saline every day. The mice in model group were given 5 ml/kg of 20% liquor every day, and the mice in Curc treatment group were treated with Curc at the doses of 5, 10, 15 mg/kg in 2 ml saline every day for 35 days. The weight of liver was measured and the health status of mice was observed. Serum ALT, AST, ALP and liver TG, TC, HDL-C, LDL-C, MDA, SOD, GSH-Px and NO were measured. The pathological changes of liver tissues stained with hematoxylin and eosin were observed. Results: Compared with the control group, the liver mass and serum levels of ALT, AST, ALP, MDA, NO, TC, TG, HDL-C, LDL-C in the model group were increased significantly (P＜0.05, P＜0.01), the activities of SOD and GSH-Px were decreased significantly (P＜0.05, P＜0.01), the liver cells were vacuolated and infiltrated with inflammatory cells, and the expression levels of NF-κB and MAPK protein in liver tissues were increased significantly (P＜0.01). Compared with the model group, the levels of ALT, AST, ALP, MDA, NO, TC, TG, HDL-C, LDL-C in Curc group were decreased significantly nd the activities of SOD and GSH-Px were increased significantly (P＜0.05, P＜0.01). Conclusion: Curc can effectively reduce liver tissue damage by regulating NF-κB/MAPK signal pathway.

Objective: To investigate the alleviating effect of hydrogen (H₂) on homocysteine (Hcy) levels and non alcoholic fatty liver in rats with hyperhomocysteinemia (HHcy). Methods: After one week of adaptive feeding, Wistar rats were randomly divided into three groups: the general diet group (CHOW), the high methionine group (HMD), and the high methionine plus hydrogen rich water group (HMD+HRW), with 8 rats in each group. The CHOW group was fed with AIN-93G feed, while the HMD and HMD+HRW groups were fed with AIN-93G+2% methionine feed to construct an HHcy model. The HMD+HRW group was also gavaged with hydrogen rich water (3 ml/animal, twice a day, with a hydrogen concentration of 0.8 mmol/L), and body weight data were recorded. After 6 weeks of feeding, the plasma and liver samples were processed and collected. The plasma homocysteine (Hcy) and lipid contents of each group were measured, and the histological morphology of the liver was observed. The activities of key enzymes in the Hcy metabolism pathway and mRNA expression were detected in the liver. Results: Compared with the CHOW group rats, the Hcy level in the blood of HMD rats was significantly increased significantly (P＜0.05). Pathological tissue sections showed liver enlargement, injury, and fatty liver in the rats; Compared with the HMD group rats, the HMD+HRW group rats showed a significant decrease in Hcy in the blood, reduced liver damage, and increased Hcy metabolism key enzyme activity and mRNA expression in the liver, with statistical differences (P＜0.05). Conclusion: Hydrogen has a significant improvement effect on liver injury induced by HMD diet in HHcy rats, possibly by enhancing the three metabolic pathways of Hcy to reduce excessive Hcy in the body, thereby improving liver metabolic function and symptoms of non-alcoholic fatty liver disease.

Objective: To investigate neuroprotective effects of total saponins from Trillium tschonoskii Maxim (TST) on vascular cognitive impairment (VCI) rats through inflammatory body of the NOD-like body protein 3 (NLRP3) regulated by endoplasmic reticulum stress (ERS). Methods: SD rats were divided into sham-operated group (SHAM), model group (VCI, bilateral neck arterial ligation (BCCAO) method), TST intervention group (TST, 100 mg/kg), and positive group (donepezil hydrochloride, 0.45 mg/kg ), continuous administration for 4 weeks. The ability of learning and memory was evaluated by the morris water labor. The tissue pathological changes were observed by HE and NISSL staining. Western blot was used to detectendoplasmic reticulum-related proteins GRP78, IRE1, XBP1. Inflammasome-related proteins NLRP3, ASC, Caspase-1, IL-18, IL-1β. Results: Compared with the SHAM group, the escape latency of VCI group rats was prolonged significantly, and the number of times of crossing the platform and the percentage of target quadrant residence time were shortened (P＜0.01); The cells in the hippocampus of VCI rats were damaged, with obvious pyknosis, decreased number of neurons and damage of cell body structure; The endoplasmic reticulum and inflammatory corpuscle-associated proteins were increased in VCI group (P＜0.05 or P＜0.01). Compared with the VCI group, the TST group and the positive group had less time to search for the platform, and the ratio of the times of crossing the platform to the time in the target quadrant was longer (P＜0.05 or P＜0.01). There was no significant difference in the times of crossing the platform between the positive group and VCI group (P＞0.05); The cell damage, nuclear pyknosis and the number of neurons in TST and positive groups were significantly reduced; The endoplasmic reticulum associated proteins and inflammatory body associated proteins in TST group and positive group were decreased to different degrees (P＜0.05 or P＜0.01). Conclusion: TST has neuroprotective effects on VCI rats, and its mechanism may be related to the involvement of ERS in the regulation of NLRP3 inflammatory small bodies.

Objective: To investigate the effects of bosutinib on the early stage of cerebral ischemia-reperfusion injury in rats. Methods: Forty Sprague-Dawley rats were randomly divided into four groups (random number method), 10 rats in each group; sham group (control group): only neck vessels were isolated without other treatments; MCAO (model group): the rat brain ischemia/reperfusion injury model was made by a modified wire bolus method,ischemia for 2 h followed by reperfusion for 24 h; DMSO group (solvent group): DMSO ( 0.752 ml/kg) was injected into the tail vein one day before the experiment, brain ischemia 2 h reperfusion for 24 h; Bosutinib group (intervention group): one day before the experiment, the tail vein was injected with Bosutinib (4 mg/kg), brain ischemia 2 h reperfusion for 24 h. After 24 h of ischemia reperfusion, neurological function score was performed; brain infarct area was calculated after staining with TTC; SIK2 was detected by Western blot; the contents of TNF-α and IL-6 in brain tissue were detected by ELISA. Results: Compared with the sham group, the neurological function scores, the infarct volume percentages and the levels of inflammatory factors IL-6 and TNF-α of the MCAO and DMSO groups were increased significantly (P＜0.05 or P＜0.01). Compared with the MCAO and DMSO groups, the above mentioned indexes of the bosutinib group were all decreased significantly (P＜0.05 or P＜ 0.01). Compared with sham group, the expression levels of SIK2 protein in MCAO and DMSO groups had no significant changes(P＞ 0.05); compared with the MCAO and DMSO group, the expression level of SIK2 protein in the bosutinib group was decreased significantly (P＜0.05). Conclusion: Bosutinib reduces cerebral ischemia-reperfusion-induced injury, and its possible mechanism is related to the decreased expression of SIK2 protein and inflammatory factors.

Objective: To investigate the effects of repeated sevoflurane exposure on hippocampal cell apoptosis and long-term learning and memory ability in neonatal rats and its regulation on PI3K/AKT pathway. Methods: Ninety SD rats were randomly divided into the control group (25% oxygen), the single exposure group (inhalation of 3% sevoflurane and 25% oxygen on the 6th day after birth), the 3-times exposure group (inhalation of 3% sevoflurane and 25% oxygen on the 6th, 7th, and 8th day after birth), the 5-times exposure group (inhalation of 3% sevoflurane and 25% oxygen on the 6th, 7th, 8th, 9th and 10th day after birth), and the 5-times exposure + 740Y-P (PI3K activator) group (intraperitoneal injection of 0.02 mg/kg 740Y-P after inhalation of sevoflurane for 5 times) according to the random number table method. Morris water maze was used to measure the learning and memory ability; HE staining and transmission electron microscopy were used to observe the morphological and structural changes of neurons in the hippocampus; TUNEL was used to detect the apoptosis of hippocampal nerve cells; Western blot was used to detect the expressions of apoptosis-related proteins (Caspase-3, Bax, Bcl-2) and PI3K/AKT pathway related proteins in the hippocampus of rats. Results: Compared with the control group and the single exposure group, the learning and memory abilities of rats in the 3-times exposure group and the 5-times exposure group were severely reduced, the morphology and structure of hippocampal neurons were severely damaged, and the apoptosis rate of hippocampal nerve cells was increased (P＜0.05), the expressions of Capase-3 and Bax proteins were significantly increased (P＜0.05), and the expressions of Bcl-2 protein and PI3K/AKT pathway protein were significantly decreased (P＜0.05). With the increase in the number of exposures to sevoflurane, the learning and memory abilities of rats were significantly reduced, the hippocampal neuron cells were severely damaged, the hippocampal neuronal apoptosis rate was significantly increased (P＜0.05), and the expressions of PI3K/AKT pathway proteins were significantly reduced (P＜0.05). Compared with the 5-times exposure group, the learning and memory abilities and hippocampal neuron structure of rats in the 5-times exposure +740Y-P group were restored to a certain extent, and the hippocampal neuronal apoptosis rate, the levels of capase-3 and Bax protein were significantly reduced (P＜0.05), while the expressions of Bcl-2 protein and PI3K/AKT pathway protein were increased significantly (P＜0.05). Conclusion: Repeated exposure to sevoflurane can significantly reduce the learning and memory abilities of neonatal rats and exacerbate hippocampal neuronal apoptosis, which may be mediated by inhibiting the PI3K/AKT pathway.

Objective: To explore the changes in the excitatory/inhibitory (E/I) balance of pyramidal neurons in prefrontal cortex and hippocampus in mice with anxiety disorder induced by chronic unpredictable mild stress (CUMS). Methods: Twenty-four C57/BL6 male mice were randomly divided into control group (CTRL) and model group (CUMS), with 12 mice in each group. The mice in CUMS group were subjected to 21 days of stress, including restraint for 1 h, reversed day/night cycle for 24 h, forced warm water bath for 5 min, water/food deprivation for 24 h, housing in wet sawdust for 18 h, shaking the cage for 30 min, noise for 1 h, and social stress for 10 min. CTRL group mice were fed normally. Anxiety-related behavioral tests and whole-cell recording tests were performed after modeling. Results: Compared with CTRL group, the time of spent in the central arena of CUMS group was reduced significantly in open field test (P＜0.01), the time and number of entering the open arms were decreased significantly in elevated plus maze test (P＜0.01), and the time of staying in the closed arms was increased significantly in CUMS group (P＜0.01). The sEPSC frequency, capacitance and E/I ratio of dlPFC, mPFC and vCA1 pyramidal neurons of mice in CUMS group were increased significantly (P＜0.01), while sEPSC amplitude, sIPSC frequency, amplitude and capacitance were not significantly changed (P＞0.05). The frequency, amplitude, capacitance and E/I ratio of sEPSC and sIPSC of dCA1 pyramidal neurons were not significantly changed (P＞0.05). Conclusion: The anxiety-like behavior of CUMS-induced mice may be the result of the participation of multiple brain regions, which is mainly related to the increase of the excitability of pyramidal neurons in dlPFC, mPFC and vCA1 brain regions, but seems to have little relationship with dCA1 brain regions.

Objective: Three modeling methods were used to establish a mouse primary liver cancer model, and compared them to find a more optimal modeling method. Methods: Forty 15-day-old C3H/HeN male mice were randomly divided into groups I-IV, 10 mice in each group. Group Ⅰ were not treated; Group Ⅱ were intraperitoneally injected with 25 mg/kg diethylnitrosamine (DEN) once; Group Ⅲ were intraperitoneally injected with 100 mg/kg DEN once; Group Ⅳ were intraperitoneally injected with 25 mg/kg DEN once and followed by another intraperitoneal injection of 100 mg/kg DEN at 42 days of age. The mortality of mice in each group was analyzed. At the 18th week of modeling, blood was collected from eyeballs after anesthesia, and liver was taken from abdominal cavity after neck was broken. The appearance of liver, the number of cancer nodules and the incidence of liver tumor were observed. The histopathological changes of liver were observed by HE staining. The serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were detected. Results: At the 18th week of modeling, compared with the group I, serum levels of ALT and AST in groups II-IV were increased significantly (P＜0.05); The number of cancer nodules and the incidence of tumors in the surviving mice of groups III and IV were also increased significantly (P＜0.05). At the 18th week of modeling, no mice died in both groups I and II, and the incidence of liver cancer was 0%; The incidence of liver cancer in surviving mice in both groups III and IV was 100%, but the mortality rate of mice in group III was as high as 50%, and that in group IV was only 20%. Conclusion: C3H/HeN male mice can successfully establish a mouse liver cancer model by intraperitoneal injection of 25 mg/kg of DEN once at the age of 15 days and another intraperitoneal injection of 100 mg/kg of DEN once at the age of 42 days with short cycle and low mortality, which is an ideal method to establish a primary liver cancer model.