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Effects of Sox11 gene on neuronal migration in the development mouse cerebral cortex
LYU Ye, SUN Xiao-wan, ZHANG Cong, LUAN Zhi-lin
CJAP    2021, 37 (4): 337-342.   DOI: 10.12047/j.cjap.6144.2021.052
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Objective: To study the effect of the schizophrenia susceptible gene Sox11 on the migration of cortical neurons using mice as experimental animals. Methods: The real-time quantitative PCR and in situ hybridization were used to clarify the expression pattern of Sox11 in the cerebral cortex during development (E14.5, P0, P7, P14). The techniques of plasmid construction, transfection, in utero electroporation and immunostaining were used to explore the role of Sox11 in the neuronal radial migration by transfecting control shRNA plasmid, mSox11 shRNA plasmid and mSox11 shRNA post-interference recovery plasmid in mice of different ages (E17.5, P0, P4, P7). Results: Compared with control neurons, the migration of mSox11 shRNA transfected neurons was delayed significantly. When a part of the neurons in the control group had reached the surface of the neocortex, most of the neurons transfected with mSox11 shRNA remained in the middle area of the neocortex. After the rat Sox11 (rSox11) gene overexpression vector was used to recuse the mouse Sox11 (mSox11) gene interference in mice, the distribution of neurons after migration was basically the same as the control. The distribution of migrating neurons in the subventricular zone (SVZ), intermediate zone (IZ), and cortical plate (CP) was different significantly (P<0.01) after Sox11 interference and recovery. Conclusion: Sox11 can promote the migration of cortical neurons, suggesting that Sox11 plays a crucial role in the migration process of mouse cortical neurons.
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Changes of lncRNA-mRNA co-expression network induced by mtDNA3010A/G genotype mutation in hypoxia
CHEN Yu, GONG Liang, LUO Yong-jun
CJAP    2021, 37 (4): 343-348.   DOI: 10.12047/j.cjap.6076.2021.038
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Objective: To analyze the long non-coding RNA (lncRNA) and messenger RNA (mRNA) co-expression network changes induced by mtDNA3010A/G mutation in acute hypoxia, and to investigate the role of key lncRNA and mRNA in the regulation of gene expression induced by hypoxia. Methods: The genotype combinations A-C-C and G-C-C of mitochondrial DNA 3010-5178-10400 were screened, and genotypes of mtDNA3010A and mtDNA3010G fusion cells were constructed by using osteosarcoma cell treated by ethidium bromide without mitochondrion (ρ0206 cell) as donors. After treatment with 1% O2 24 h, the lncRNA - mRNA expression chip was applied to detect the differently expressed lncRNA and mRNA in two kinds of fusing cells, and fluorescence quantitative polymerase chain method was used to verify differently expressed mRNA. Bioinformatics methods were applied to build co-expression network of lncRNA-mRNA, predict target genes of differently expressed lncRNA, and the functions of differently expressed mRNA and target genes predicted by lncRNA were also analyzed based on gene ontology (GO) and the Kyoto encyclopedia of genes and genomes (KEGG) forecast analysis. Results: After treatment with 1% O2 for 24 h, compared with mtDNA3010G fusion cells: 688 lncRNAs were up-regulated, 21 were more than 2 times; 1098 were down-regulated, and 4 were more than 2 times. There were 1151 mRNA expressions up-regulated, 14 were more than 2 times, 539 mRNA expressions were down-regulated, and 3 were more than 2 times. Conclusion: MtDNA3010A/G genotype mutation under hypoxia is able to affect the lncRNA-mRNA regulatory network, and the differentially expressed lncRNA and mRNA may play an important role in regulation network of gene expression induced by hypoxia, which is expected to be a target for the regulation of hypoxia reaction from the perspective of mitochondria.
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Effects of Notch3 on gene expression and signal pathway of pancreatic stellate cell activation
SONG Hai-yan, ZHOU Zhi-xin, ZHANG Yu-xiang
CJAP    2021, 37 (4): 349-353.   DOI: 10.12047/j.cjap.6084.2021.024
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Objective: The pancreatic stellate cells( PSCs) of mice were isolated and cultured and the effects of Notch3 siRNA on PSCs gene expression were detected. Methods: were PSCs of mice were isolated and cultured. The expressions of α-SMA, fibonectin and collagen I in activated PSCs were detected by immunofluorescence. The PSCs were divided into four groups, blank control group (MOCK group), negative siRNA control group (NC group) , Notch3 siRNA group (N3 siRNA group) and Notch3 siRNA-1 group (N3 siRNA-1 group). Cell treatment: the same transfection method was applied to transfect PSCs for 48h. Then total RNA was extracted from each group, and the concentration and purity of RNA was measured. The transcriptome sequencing and analysis were performed by ANOROAD Gene technology (Beijing) co., LTD. Results: The immunofluorescence results showed that α-SMA, fibonectin and collagen I were significantly expressed in activated PSCs. The analysis of the sequencing results showed that the gene expressions of α-SMA, collagen I, fibronectin, CTGF and PCNA of PSCs were down-regulated in N3 siRNA group and N3 siRNA-1 group comparing with NC group. The genes involved in collagen metabolism were up-regulated, the gene expression of positive regulation of collagen biosynthesis was down-regulated, while that of negative regulation of collagen biosynthesis was up-regulated in N3 siRNA group and N3 siRNA-1 group comparing with NC group. The genes that regulated cell aggregation were down-regulated, and the genes that regulated extracellular matrix were down regulated in N3 siRNA group and N3 siRNA-1 group comparing with NC group. Inhibition of Notch3 expression in PSCs could affect the gene expressions of cell adhesion molecule signaling pathway, MAPK signaling pathway and TGF-β signaling pathway. Conclusion: Inhibition of Notch3 expression can inhibit activation of PSCs , and reduce the ability of proliferation, migration and aggregation and ECM synthesis of PSCs. Inhibition of Notch3 expression may affect other signal pathways such as cell adhesion molecule signaling pathway, MAPK signaling pathway and TGF-β signaling pathway, but its effects need further validation.
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Effects of GPR81 agonist on insulin resistance in rats with nonalcoholic fatty liver disease
ZHANG Yu, LU Qing-hua, CAO Hai-fang, ZHANG Sheng-rong, WANG Hu-de
CJAP    2021, 37 (4): 354-358.   DOI: 10.12047/j.cjap.6055.2021.033
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Objective: To investigate the effects of NOD-like receptor protein 3 (NLRP3) signaling pathway on insulin resistance and the intervention of lactic acid receptor G protein-coupled receptor 81 (GPR81) agonist in nonalcoholic fatty liver disease (NAFLD) rats. Methods: Thirty SD male rats were randomly divided into three groups: control group, NAFLD group and GPR81 agonist group, with 10 rats in each group. Nonalcoholic fatty liver rat model was established by high fat diet. The rats in GPR81 agonist group were injected intraperitoneally with GPR81 specific agonist lactate (50 nmol/L) on the basis of nonalcoholic fatty liver model once a week, and the other two groups were injected with the same amount of normal saline for 12 weeks. The levels of liver biochemical indexes, fasting blood glucose, insulin and inflammatory factors in liver homogenate were measured, and the histopathological morphology of liver in each group was observed. The protein expressions of NLRP3, apoptosis-associated speck-like protein containing a CARD (ASC), cysteinyl aspartate specific proteinase-1 (caspase-1), insulin receptor substrate 1 (IRS-1), Tyr465-IRS-1, Ser636-IRS-1, glucose transporter 4 (GLUT4) in liver tissue were detected by Western blot. The mRNA expression levels of NLRP3, ASC, caspase-1, IRS-1 and GLUT4 in liver tissue were detected by qRT-PCR. Results: Compared with the control group, the serum levels of triglyceride (TG), alanine aminotransfease (ALT), aspartate aminotransfease (AST), fasting plasma glucose (FPG), fasting insulin (FINS) and homeostasis model assessment of insulin resistance (HOMA-IR) of NAFLD group were increased significantly (P< 0.05). The results of liver histopathology showed that in NAFLD group, there were obvious fatty changes in liver tissue, fat droplets in hepatocytes and inflammatory cell infiltration. And the mRNA and protein expressions of NLRP3, ASC, caspase-1 and the protein expression of Ser636-IRS-1 in NAFLD group were increased significantly, and the contents of interleukin 1β (IL-1β) and interleukin 18 (IL-18) in liver and serum were increased, while the mRNA and protein expressions of IRS-1 and GLUT4 and Tyr465-IRS-1 were decreased significantly (P<0.05). Compared with NAFLD group, the above indexes of GPR81 agonist group were all significantly improved. Conclusion: The activation of NLRP3 signaling pathway mediates the production of inflammatory factors and promotes the development of NAFLD. GPR81 agonist may be a potential treatment for NAFLD.
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Effects of acupuncture on endoplasmic reticulum pathway in exercise-induced skeletal muscle damage in rats and its mechanism
DING Hai-li, HUANG Zeng-hao, REN Zai-fang, SUN Zhu-xin, LI Jun-ping, WANG Rui-yuan
CJAP    2021, 37 (4): 359-364.   DOI: 10.12047/j.cjap.6097.2021.044
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Objective: To observe the effects of acupuncture on the endoplasmic reticulum (ER) functional enzymes sarcoplasmic reticulum Ca2+-ATPase (SERCA), protein disulfide isomerase (PDI), glucose-regulated protein 78(GRP78) and PERK pathways in rats with exercise-induced skeletal muscle damage, and to explore the mechanisms of acupuncture in preventing and treating exercise-induced skeletal muscle damage. Methods: Eight-week-old male SD rats were randomly divided into control group (group C, n=6), exercise group (group E, n=30), acupuncture group (group A, n=30) and exercise acupuncture group (group EA, n=30). Among them, the E and EA group were established an exercise-induced skeletal muscle damage model by a single eccentric exercise, and acupuncture intervention was applied 0.5 cm above the Achilles tendon of the rat's calf immediately after EA exercise, and in group A, acupuncture intervention was applied during the same period. Each group was divided into 0 h/12 h/24 h/48 h/72 h (n=6) according to different sampling time points after exercise and acupuncture intervention, and soleus muscle was collected at the corresponding time for index test. The ultrastructure of muscle fibers was observed by transmission electron microscopy; the contents of SERCA and PDI were determined by ELISA; and the expressions of ER stress marker proteins GRP78 and p-PERK and p-eIF2α were detected by Western blot. Results: Compared with group C, there were no significant differences in the indicators of group A at all time points (P> 0.05), the ultrastructure of muscle fibers in group E showed different damages, SERCA content was significantly decreased from 0 h to 48 h (P<0.05), PDI content was significantly increased from 0 h to 72 h (P<0.05), GRP78 expression was significantly increased from 0 h to 72 h (P<0.05), p-PERK expression was significantly increased from 0 h to 24 h (P<0.05), and p-eIF2α expression was consistent with p-PERK. Compared with the corresponding times in group E, the ultrastructure of muscle fibers in group EA was significantly alleviated, SERCA content was significantly increased from 48 h and 72 h (P<0.05), PDI content was significantly increased from 0 h to 72 h (P<0.05), and GRP78 expression was significantly decreased from 0 h to 72 h (P<0.05). Conclusion: Acupuncture can effectively ameliorate exercise-induced skeletal muscle damage and alleviate ER stress after a large load eccentric exercise. The mechanism of them may be related to the up-regulation of protein disulfide isomerase PDI and the inhibition of ER stress PERK pathway.
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Effects of electroacupuncture on autophagy factors and liver lipid metabolism in rapidly aging mice
YANG Yun- hao, PANG Fang, HUANG Si-qin, YANG Zhi-xue, ZHU Zheng-wei, DAI Pan, GUO Xiao, LIAO Dong-mei, TANG Cheng-lin
CJAP    2021, 37 (4): 365-370.   DOI: 10.12047/j.cjap.6156.2021.053
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Objective: To study the effects of electroacupuncture on the expressions of autophagy-related factors LC3-Ⅱ, Beclin1, Atg7, and P62 in the liver of rapidly aging (senescence accelerated mouse/prone8,SAMP8) mice, and to explore the mechanisms of electroacupuncture to improve liver lipid metabolism in mice. Methods: Thirty-week-old male SAMP8 mice were randomly divided into model group, drug group, and electroacupuncture group, with 7 mice in each group. Seven anti-rapid aging SAMR1 mice of the same age were used as the control group. The animals in the control group and the model group were bred routinely for 2 weeks without any intervention; the drug group was treated with intraperitoneal injection of rapamycin at the dose of 10 mg·kg-1·d-1, once a day, 6 consecutive days a week; the electroacupuncture group was given "Shenshu" and "Taichong" Electroacupuncture at point(15 minutes a day, 6 consecutive days a week). The serum lipid metabolism and liver lipid deposition of mice were detected, the distribution of liver autophagy body, the protein and mRNA expressions of liver LC3 - Ⅱ, Beclin1, Atg7 and P62 were determined. Results: Compared with the control group, the total cholesterol (TC), triglycerides (TG), and low density lipoprotein (LDL) of the model group were increased significantly(P<0.01). In the model group, lipid droplet deposition was obvious, autophagosomes were decreased, the protein and mRNA expression levels of autophagy- related factors LC3-Ⅱ, Beclin1 and Atg7 were decreased significantly (P<0.01), while the protein and mRNA expressions of P62 were increased significantly (P<0.01). Compared with the model group, the serum contents of TG, TC, and LDL of the mice in the electroacupuncture group and the drug group were decreased significantly (P<0.01), lipid droplet deposition was reduced, autophagosomes were increased, the protein and mRNA expression levels of LC3 -Ⅱ, Beclin1 and Atg7 were increased significantly(P<0.01), and the protein and mRNA expression levels of P62 were decreased significantly(P<0.01). The protein and mRNA expression levels of Beclin1 and Atg7 in the liver of the electroacupuncture group were not significantly different from the drug group (P>0.05). Conclusion: Electroacupuncture can alleviate liver lipid metabolism disorders, which may be related to the regulation of the expressions of liver autophagy related factors LC3-Ⅱ, Beclin1, Atg7, and P62, thereby promoting liver autophagy in SAMP8 mice.
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Characteristics of circulation and microcirculation in healthy people of Han nationality at different altitudes
HE Zong-zhao, DENG Li, MA Si-qing, LI Xin-hui, WANG Hao
CJAP    2021, 37 (4): 371-375.   DOI: 10.12047/j.cjap.6014.2021.006
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Objective: To observe the circulation and microcirculation characteristics of healthy han people in Qinghai at different Guoluo (4 100 m) and Huzhu (2 600 m) and in Shanghai in plain area (4 m). Methods: The 28 healthy han people aged (45.62±10.15) from Guoluo in Qinghai, 27 healthy han people aged (47.25±8.43) from Huzhu in Qinghai and 20 healthy han people aged (43.12±8.28) from Shanghai were divided into three groups: Guoluo group (GL), Huzhu group (HZ) and Shanghai group (SH); Venous blood was collected for routine blood test: red blood cells count(RBC), hemoglobin(HGB), hematocrit(HCT), platelet(PLT); and coagulation index: prothrombin time(PT), interntionl normlized ratio(INR), fibrin(Fib), activated partial thromboplastin time (APTT), thrombin time(TT), continuous non - invasive hemodynamic monitor (CNAP) was used for continuous measurement: cardiac output (CO), heart rate (HR), stroke volume (SV), pulse pressure variation rate (PPV), systemic vascular resistance index (SVRI), mean arterial pressure (MAP); pulse oxygen monitoring instrument is used for monitoring SPO2 (pulse oxygen); the total vascular density (TVD), perfusion vascular density (PVD), proportion of perfused vascular(PPV), and microvascular flow index (MFI) of sublingual microcirculation were observed by using peripheral dark field imaging (SDF) monitoring technique. Results: Compared with the Shanghai group, the RBC and HGB levels in the Huzhu group and the Guoluo group were significantly increased (P<0.05), while PLT levels were significantly decreased (P<0.05); The levels of PT, Fib, APTT, TT, CO, HR, SV, MAP and SPO2 in the Huzhu group were significantly decreased (P<0.05), while the levels of TVD, PVD, PPV and MFI were significantly increased (P<0.05). In Guoluo group, the levels of HCT, Fib, CO, SV, PPV, MAP, TVD and PVD were significantly increased (P<0.05), while the levels of PT, INR, TT and SPO2 were significantly decreased (P<0.05). Compared with the Huzhu group, the levels of RBC, HGB, HCT, Fib and APTT of Guoluo group were significantly increased (P<0.05); while the levels of SPO2, TVD, PVD, INR and TT were significantly decreased (P<0.05). Conclusion: The microcirculation of healthy people in different altitude areas is significantly different, and the microcirculation density of healthy people in high altitude areas is significantly higher than that in plain areas, especially in moderate altitude areas. Its special physiological significance is of guiding significance for the treatment of infectious shock and chronic altitude disease in plateau areas.
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CJAP    2021, 37 (4): 376-379.   DOI: 10.12047/j.cjap.6022.2021.009
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Effects of Mongolian medicine Shaosha-7 on myocardial ischemia/ reperfusion injury of rats
YANG Xiang-jun, WANG Yu, E.bayarima, ZHAO Ming, S. hexigezarigala
CJAP    2021, 37 (4): 380-384.   DOI: 10.12047/j.cjap.6114.2021.029
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Objective: To investigate the effects of Shaosha-7 in rats with myocardial ischemia-reperfusion injury and its mechanisms. Methods: male SD rats were divided into sham operation group (n=10), myocardial ischemia-reperfusion injury group (n=10), low, medium and high dose of Shaosha-7 groups (n=10), and positive drug group (n=10). The rats of Shaosha-7 (low, medium and high dose) groups were treated with Shaosha-7 at the doses of 0.4, 0.8 and 1.6 g/kg respectively, once a day for 15 days. The rats of positive drug group were treated with 0.3 g/kg Danshen, once a day for 15 days. The rats of the sham operation group and myocardial ischemia-reperfusion injury were treated with 2 ml/100 g distilled water, once a day for 15 days. After 15 days, the rats of the model group and the treatment group underwent thoracotomy and ligation of coronary artery for 30 minutes, then thoracic cavity was closed after reperfusion. Rats in six groups were executed electrocardiographic examination and their hearts were taken for Hematoxylin-Eosin (HE) staining and 2, 3, 5-triphenyltetrazolium chloride (TTC) staining to observe infarct area and myocardial pathological changes. The contents of cTnI, CK-MB, LDH, MDA, SOD, GSH-Px, TNF-α, IL-18, IL-6 and IL-1 β in serum were detected by ELISA. The expression of NF-кB was detected by immunohistochemistry. Results: Compared with the sham operation group, the infarct size, the levels of cTnI, CK-MB, CK-MB, LDH, MDA, GSH-Px, TNF-α, IL-18, IL-6, IL-1β and NF-кB were increased and the content of SOD were decreased in rats with myocardial ischemia-reperfusion injury. Compared with the rats with myocardial ischemia-reperfusion injury, Shaosha-7 improved the arrhythmia and pathological changes, reduced the infarct area, decreased the contents of cTnI, CK-MB, LDH, MDA, GSH-Px, TNF-α, IL-18, IL-6, IL-1 β, increased the content of SOD, decreased the expression of NF-кB. Conclusion: Mongolian medicine Shaosha-7 can effectively alleviate myocardial ischemia-reperfusion injury in rats. This study provides a theoretical basis for the treatment of myocardial ischemia-reperfusion (I/R) injury with Shaosha-7.
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The regulatory role of autophagy in rats lung ischemia/reperfusion injury
HAO Mao-lin, LOU Guo-qiang, LIU Xiu-jie, QIAN Wei, WANG Jia, ZHOU Zhuo-lin, WANG Wan-tie
CJAP    2021, 37 (4): 385-388.   DOI: 10.12047/j.cjap.6061.2021.019
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Objective: To investigate the role of cell autophagy in lung ischemia/reperfusion injury in rats. Methods: Forty SD rats were randomly divided into 5 groups (n=8): ①Sham operated group (sham group):just open rat chest for 3.5 h; ②Ischemia/reperfusion group (I/R group):after open chest, clamp pulmonary hilus for 0.5h then reperfusion for 3 h; ③Solvent group (DMSO group): intraperitoneal injection of DMSO solution for 1h before operation; ④Autophagic inhibitor group (3-MA group); ⑤Autophagic agonist group (Rap group): intraperitoneal injection of autophagic agonist rapamycin before operation; the rest operations of DMSO, 3-MA and Rap groups are the same as that of I/R group. At the end of the experiment, the rats were killed by euthanasia-killing. The lung tissues were collected and the wet/dry weight ratio (W/D) and total lung water content (TLW) of the lung tissues were detected. The lung tissue structure and cell ultramicro morphology were observed by light microscopy and electron microscopy and the injuried alveolar rate(IAR) was calculated. The autophagy-related protein expressions were detected by Western blot. Results: Compared with sham group, the levels of W/D, TLW and IAR were increased, the expressions of autophagy related protein and p-AMPK, Beclin 1, LC3 II were also increased in other four groups, while the protein expressions of p-mTOR and p62 were decreased significantly (P< 0.05 or P<0.01). Under the light microscope, the other groups of lung tissue had edema and exudation in varying degrees, the structure of alveoli was disordered, the ultrastructural damage of cells was aggravated under the electron microscope, and autophagosome could be observed. Compared with DMSO group, the expressions of autophagy related protein, the levels of W/D, TLW and IAR in 3-MA group were decreased (P<0.05 or P<0.01), the edema of lung interstitial was lighter, and less cells were found in alveolar cavity. Ultrastructural damage was also lighter and with less autophagosome. Besides, there was no significant difference among I/R, DMSO and Rap groups (P>0.05). Conclusion: Autophagy can be activated during ischemia/reperfusion in rats to induce lung injury.
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CJAP    2021, 37 (4): 389-392.   DOI: 10.12047/j.cjap.6027.2021.030
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Changes of FLI-1 protein expression in mice with pulmonary endothelial barrier dysfunction following acute lung injury induced by lipopolysaccharide
XU Li-cong, HUANG Ting-ting, HAO Jia-le, FAN Xiao-fang, GONG Yong-sheng, MAO Sun-zhong
CJAP    2021, 37 (4): 393-396.   DOI: 10.12047/j.cjap.6080.2021.040
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Objective: To observe changes of Friend leukemia virus integration 1 (FLI-1) protein expression of pulmonary tissue in mice with pulmonary endothelial barrier dysfunction following acute lung injury (ALI) induced by lipopolysaccharide (LPS). Methods: The mouse model of ALI was established by injection of LPS (7.5 mg/kg, i.p. ). At 0 h, 12 h, 24 h and 48 h after LPS injection, pulmonary microvascular endothelial permeability and lung wet/dry weight ratio (W/D) were assessed. The contents of TNF-α and IL-6 in bronchoalveolar lavage fluid (BALF) were detected by ELISA method. The protein levels of FLI-1 and Src protein tyrosine kinase (SRC) were analyzed by Western blotting.Results: ①Pulmonary microvascular endothelial permeability at 12 h and 24 h were significantly higher than those of 0 h by 74.3% and 162.4%, respectively, while that of 48 h was lower than that of 24 h by 27.0% (P<0.05). The W/D at 12 h and 24 h were significantly higher than those of 0 h by 50.1% and 122.9%, respectively, while that of 48 h was lower than that of 24 h by 10.7% (P<0.05). ②The contents of IL-6 and TNF-α in BALF at 12 h and 24 h were significantly higher than those of 0 h, while those of 48 h were significantly lower than those of 24 h by 28.3% and 21.6% (P<0.05), respectively. ③The protein level of FLI-1 in lung at 12 h and 24 h were down-regulated than those of 0 h by 20.4% and 56.9%, respectively, while that of 48 h was up-regulated than that of 24 h by 18.2% (P<0.05). The protein level of SRC in lung at 12 h and 24 h were up-regulated than those of 0 h by 76.8% and 176.7%, respectively, while that of 48 h was down-regulated than that of 24 h by 33.4% (P<0.05).④Same as the protein level of FLI-1 with the protein level of SRC in lung, pulmonary microvascular endothelial permeability was significantly negative correlated with the protein level of FLI-1 in lung, while it was significantly positive correlated with the protein level of SRC (P<0.01). Conclusion: FLI-1 participates in the pathological proceeding of pulmonary endothelial barrier dysfunction following ALI induced by LPS.
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Protective effects of total saponins of Codonopsis on ulcerative colitis induced by TNBS in rats and its mechanism
LIU Xue-feng, QIAO Jing, GAO Jian-de, CHEN Zheng-jun, LIU Xiong
CJAP    2021, 37 (4): 397-401.   DOI: 10.12047/j.cjap.6051.2021.032
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Objective: To study the protective effects and mechanisms of total saponins of Codonopsis (TSC) on ulcerative colitis in rats. Methods: Fifty male Wistar rats were randomly divided into 5 groups: control group, model group, salazosulfadiazine (SASP) positive control group (0.3 g/kg), TSC high- and low-dose experimental groups(1.2, 0.4 g/kg). UC rat model was established by trinitrobenzene sulfonic acid (TNBS)/ ethanol enema. After administration for 21 days, the rats' symptoms and signs, disease activity index (DAI), colonic mucosal injury index (CMDI) and colonic tissue morphology were observed. The contents of superoxide dismutase (SOD), malondialdehyde (MDA), inflammatory cytokines interleukin-6 (IL-6), interleukin-10 (IL-10) and tumor necrosis factor (TNF-α) in colon tissues were determined. Protein expression of nuclear nuclear transcription factor-κB (NF-κB) in colon tissues was detected. Finally, the effect of TCS therapy was evaluated. Results: Compared with the control group, the DAI and CMDI scores of the rats in the model group were increased significantly, meanwhile the colonic mucosa was seriously damaged, indicating that the model was successful. Compared with the model group, the TSC high and low dose groups could significantly reduce the DAI and CMDI score (P<0.05) and improve the colonic mucosa form. TSC also could increase the SOD activity and decrease MDA content in colon tissues(P<0.05), while inhibit the levels of IL-6 and TNF-α mRNA in the colon tissues and promote the expression of IL-10 mRNA (P<0.01). At the same time, TSC reduced the expressions of NF-κB protein in the colon (P<0.01). The TSC high-dose group was superior to the low-dose group (P<0.05). Conclusion: TSC has significant protective effects on ulcerative colonic mucosal damage in UC rats, and there is a dose-dependent relationship; its mechanism may be related to anti-lipid peroxidation and inhibiting the NF-κB signaling pathway to regulate the release of inflammatory factors.
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Effects of miR-335-5p targeting G6PD on proliferation and apoptosis of colon cancer cells
FAN Xiao-qun, LI Huan, ZHU Hua-xiong, HUANG Jian-ping, HE Lyu-fen
CJAP    2021, 37 (4): 402-406.   DOI: 10.12047/j.cjap.6090.2021.041
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Objective: To investigate the effects of miR-335-5p targeting glucose-6-phosphate dehydrogenase (G6PD) on the proliferation and apoptosis of colon cancer cells. Methods: Normal colon cell group, blank control group, NC group and miRNA-335-5p mimic group were set up. Colonic epithelial cells (IEC) and human colon cancer cells SW480 were cultured in vitro, and the cells in the NC group and miRNA-335-5p mimic group cells were transfected. RT-qPCR was used to detect the expression levels of miR-335-5p and G6PD mRNA in each group of cells. The targeting effect of miR-335-5p on G6PD was verified by Double Luciferase Report experiment. MTT assay was used to detect cell proliferation. Flow cytometry was used to detect the apoptosis rate. The expressions of G6PD, Bax, Bcl-2 and caspase-3 were detected by Western blot. Results: Compared with normal colon cells, the relative expression levels of miR-335-5p in SW480 cells of colon cancer in the blank control group and NC group were decreased, and the relative expression level of G6PD mRNA was increased (P<0.05); compared with the blank control group and NC group, the expression level of miR-335-5p in miR-335-5p mimic group was increased significantly, and the expression of G6PD mRNA was decreased significantly (P<0.05). Compared with the blank control group and NC group, the proliferative activity of colon cancer SW480 cells in miR-335-5p mimic group was decreased significantly, and the apoptosis rate was increased significantly (P<0.05). The relative activity of luciferase in miR-335-5p mimic + WT-G6PD 3 '- UTR group was lower than that in miR-335-5p NC + WT-G6PD 3' - UTR group (P<0.05). Compared with the blank control group, the relative expression levels of G6PD and bcl-2 protein in miR-335-5p mimic group were decreased significantly, and the expression levels of Bax and caspase-3 protein were increased significantly (P<0.05). Conclusion: MiR-335-5p may inhibit the proliferation and promote apoptosis of colon cancer cells by targeting G6PD.
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Clinical and prognostic features of the patients with esophageal squamous cell carcinomas as a first primary malignancy
WEN Zhen, ZHANG Yan-qiu, WU Rong, PENG Xue-mei, CHEN Bin, LENG Ai-min
CJAP    2021, 37 (4): 407-414.   DOI: 10.12047/j.cjap.6020.2021.008
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Objective: To retrospectively analyze the clinical characters and prognosis of the patients with Esophageal Squamous Cell Carcinomas as First Primary Malignancy (ESCCFPM), which will help us better understand the relationship between Esophageal Squamous Cell Carcinoma (ESCC) and other cancers, and to provide appropriate research evidence for the clinical diagnosis and treatment. Methods: The clinicopathological and follow-up data of 540 Patients with ESCCFPM between January 1, 2004 and December 31, 2016 were collected from the Surveillance, Epidemiology and End Results (SEER) database of National Cancer Institute. The Kaplan-Meier method was used to determine Overall Survival (OS) curves of ESCC patients, and the Log-Rank test was used to estimate differences in survival. The Cox proportional hazards models were adopted for the prognosis analyses. Results: Regarding the number of multiple primary malignancies (MPMs), 491 had two malignancies, 42 had three malignancies and 7 had four malignancies. ESCCFPM is more common among males. The high incidence age is between 61 and 80 years old. Tumors of the respiratory system (36.9%), were the most common MPMs followed by digestive system (35.2%) and reproductive system (8.9%). The 1-year, 3-year, 5-year OS rates for patients with ESCCFPM were 76.9%, 50.4% and 38.9%, respectively. The age of the ESCC diagnosed, T stage, time of occurrence, carcinoma number, lymph node dissection, surgery, radiotherapy and chemotherapy were the prognostic factor of overall survival for ESCCFPM patients. Age, race, T stage, time of occurrence surgery and radiotherapy were independent prognostic factors for the whole cohort by multivariate survival analysis. Conclusion: ESCCFPM,mainly two-lesion cancer, is most commonly found in respiratory system and digestive systems. Enhanced follow-up of respiratory and digestive tumors in ESCCFPM patients aged 61-80 may help identify multiple primary malignancies. Surgery, radiotherapy and chemotherapy may improve overall survival for ESCCFPM patients.
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The effects of shikonin on liver cancer cells SMMC-7721 apoptosis and its mechanism
WANG Yu, HE Shi-yan, ZHU Ruo-ting, KE Rui-jun, CHEN Jia-yu
CJAP    2021, 37 (4): 415-418.   DOI: 10.12047/j.cjap.6137.2021.051
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Objective: To investigate the effects and molecular mechanisms of shikonin on liver cancer SMMC-772 cells. Methods: SMMC-7721 cells were treated with shikonin at the concentrations of 0, 5, 20, 80 and 320 ng/ml for 0, 24, 48 and 72 h respectively. The proliferative activity of the cells was detected by CCK8 assay. The nuclear type changes of cells was observed after hoechst 33342 staining. Flow cytometry was used to analyze cell apoptosis and death rate. The expressions of proteins in cells were determined by Western blot, and the tumor inhibitive effects were observed through anti-tumor experiment on the BALB/c mice. Results: In vitro experiments, shikonin could inhibit the proliferation of SMMC-7721 cells and induce their apoptosis(P<0.01), up-regulate the expression of p53 gene, down-regulate the phosphorylation levels of AKT and PI3K protein. In vivo study also confirmed that shikonin could significantly inhibit the growth of tumor in tumor-bearing mice(P<0.01)in dose-dependent and time-dependent manners. Conclusion: Shikonin can inhibit the proliferation activitity and induce apoptosis of SMMC-7721 cells by affecting the PI3K/AKT signal pathway and has potential anti-liver cancer functions.
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Effects of betulinic acid on apoptosis of human gastric cancer MGC-803 cells
FAN Hua, SHAO Shu-li, HE Meng-qi, HUANG Xin , ZHANG Wei-wei, ZHANG Zhen-zhu
CJAP    2021, 37 (4): 419-422.   DOI: 10.12047/j.cjap.6078.2021.039
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Objective: The effects of betulinic acid (BA) on apoptosis of human gastric cancer MGC-803 cells was investigated by using human gastric cancer MGC-803 cells as experimental materials, and the basis for its clinical application was provided. Methods: The human gastric cancer MGC-803 cells were divided into 4 groups,each group was set with 3 replicates.The control group was MGC-803 cells without being added betulinic acid; the other 3 groups of experimental groups were treated with betulinic acid at final concentrations of 10, 20 and 30 μg /ml respectively. Cells were treated with betulinic acid of different concentrations for 48 h. Laser confocal microscope was used to observe morphological changes of MGC-803. The activities of Caspase-3 and Caspase-9 were detected by an assay kit. Flow cytometry was applied to determine mitochondrial membrane potential. The mRNA and protein levels of Caspase-3, Caspase-9 and Cyt c were also detected by qRT-PCR and Western blot, respectively. Results: Compared with the control group, the activities of Caspase-3 and caspase-9 were increased(P<0.01), while the mitochondrial membrane potential was decreased significantly(P<0.01). The mRNA and protein expressions of Caspase-3, caspase-9 and Cyt c were up-regulated significantly(P<0.01). Conclusion: In the final concentration range of 10 ~ 30 μg/ml, betulinic acid can induce apoptosis of human gastric cancer MGC-803 cells by regulating the expression of Caspase-3, Caspase-9 and Cyt c.
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Progranulin attenuated asthma inflammation by inhibiting the expression of IL-6
QI Yu-hong, SUN Si, WANG Ling, YANG Jing, WU Guang-ying, HUANG Qi, XU Wen-chun
CJAP    2021, 37 (4): 423-428.   DOI: 10.12047/j.cjap.6126.2021.050
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Objective: To investigate the role and mechanism of progranulin (PGRN) in asthma. Methods: Control group and model group were set up in wild and IL-6 knockout (IL-6 ko) mice, respectively. For asthma model, mice were intraperitoneally sensitized with 100 μg OVA on days 0 and 7, followed by aerosol challenges with 5% OVA for 30 min per day from day 14 to 21, and mice were sacrificed 24 h after the last challenge. The mice in control group were treated in the same way with PBS. Bronchoalveolar lavage fluid (BALF) was collected for leukocytes count and differential count. The pathological changes of lung tissues were observed by H&E staining. The cytokines in lung homogenate, serum and BALF were detected by Q-PCR and ELISA. The in vitro model of asthma was induced by stimulating A549 or BEAS-2B cells with IL-13. Each group was replicated in three wells and four groups were designed: PBS group, IL-13 treatment group, IL-13 + rhPGRN treatment group, inhibitors of p38 phosphorylation (SB203508) treatment group. The cells or supernatant were collected after 0~48 h. PGRN and IL-6 levels were determined by Q-PCR and ELISA, the level of p38 phosphorylation was tested by Western blot (WB). Results: Compared with control group, PGRN levels were decreased in lung homogenate and BALF (P<0.05), and PGRN presented a downtrend in serum, however, the level of IL-6 in BALF was increased in asthma mice (P<0.01). In IL-6 ko asthma mice, compared with the wild asthma mice, leukocytes, especially neutrophils in BALF were decreased (P<0.05), but PGRN was increased (P<0.05), lung pathological damage was significantly alleviated. In vitro experiments, compared with PBS group, PGRN level was decreased (P<0.05), IL-6 level was increased (P<0.01), phosphorylation of p38 was activated in IL-13 treatment group. Compared with IL-13 treatment group, in IL-13 + PGRN treatment group, IL-6 level was decreased (P<0.05); phosphorylation of p38 was inhibited (P<0.05); and the production of IL-6 (P<0.05) was decreased after treatment with inhibitor of p38 phosphorylation. Conclusion: PGRN inhibited the production of IL-6 by suppressing the p38 phosphorylation to alleviate asthmatic airway inflammation.
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CJAP    2021, 37 (4): 429-432.   DOI: 10.12047/j.cjap.6121.2021.047
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Effects of astaxanthin combined with aerobic exercise on renal aging of rat induced by D-galactose and its mechanism
NIU Yan-long, CAO Jian-min, ZHOU Qi-yun, HU Ge, GUO Xian, LIU Jian, HAO Min, ZHANG Tao, ZHOU Hai-tao
CJAP    2021, 37 (4): 433-438.   DOI: 10.12047/j.cjap.6064.2021.034
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Objective: To study the effects and mechanisms of astaxanthin combined with aerobic exercise on renal senescence of rat induced by D-galactose. Methods: Sixty 3-month-old SPF SD rats were divided into control group (C group), acute senescence group (S group), astaxanthin+acute senescence group (AS group), aerobic exercise+acute senescence group (ES group), astaxanthin+aerobic exercise+acute senescence group (AES group), by two-factor two-level 2×2 factorial design with 12 rats in each group. Acute senescence model of rat was establshed by intraperitoneal injection with 100 mg/(kg·d) D-galactose, and the intervention was conducted with 20 mg/(kg·d) astaxanthin and/or aerobic exercise with 60% VO2max for 6 weeks. The histopathological/ultrastructural changes of the kidney were observed by light microscope/electron microscope; the levels of SOD, γ-GCS and MDA were detected by ELISA, and LDF in kidney was determined by fluorescence colorimetry; the protein expression of Nrf2 signaling pathway was detected by immunohistochemistry. Results: Compared with AS and ES group, in AES group, the improvement of renal tissue morphology/ultrastructure was more significant; LDF was decreased significantly (P<0.01); SOD activity was significantly increased (P<0.01); γ-GCS was significantly higher than that of AS group, but not significantly different from that of ES group (P>0.05); there was no significant difference in MDA between groups (P>0.05); the levels of Nrf2 and p-Nrf2 were increased significantly (P<0.05, P<0.01); HO-1 was significantly higher than that of ES group(P<0.05), but not significantly different compared with that of AS group(P>0.05). Conclusion: Astaxanthin combined with aerobic exercise can delay aging process of kidney, its mechanism may be that the combination regulate the protein expression in Nrf2 signaling pathway, Ⅱ detoxifying enzymes and antioxidant enzyme activity, and improve oxidative stress in kidney of rat induced by D-galactose.
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Effects of aerobic interval training on myocardial oxidative stress and inflammation in rats with myocardial infarction and its mechanism
GENG Yuan-wen, LIN Qin-qin, WANG Xiang-yi, LI Ruo-ming, TIAN Zhen-jun
CJAP    2021, 37 (4): 439-444.   DOI: 10.12047/j.cjap.6094.2021.043
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Objective: The present study was to determine the effects of aerobic interval training (AIT) on the expressions of SIRT1, Nox4 and inflammatory factor in the heart of rats with myocardial infarction (MI). Methods: Male Sprague Dawley rats were randomly divided into sham-operated group (C), sedentary MI group (MI) and MI with AIT group (ME) (n=10). The MI model was established by ligation the left anterior descending coronary artery. Rats in C groups were subjected to the same surgery, but only threaded and not ligated. After surgery 1 week, rats in ME groups took adaptability training for 1 week, and then subjected to 4 weeks treadmill exercise training. After training, the hearts were collected for histological observation. The level of malondialdehyde (MDA) and the activity of superoxide dismutase (SOD) in heart was assessed by ultraviolet spectrophotometry. The activity of lactate dehydrogenase (LDH) was determined by enzyme linked immunosorbent assay. The expression of sirtuin1 (SIRT1) mRNA was examined by real-time quantitative polymerase chain reaction (RT-qPCR). The protein expressions of SIRT1, NADPH oxidase 4 (Nox4), tumor necrosis factor-alpha (TNF-α) and interleukin-1β (IL-1β) were detected by Western blotting. The levels of reactive oxygen species (ROS) were detected by dihydroethidium (DHE) staining. Results: Compared with the C group, the expression level of cardiac Nox4 protein was increased (P<0.01), the level of MDA, activity of LDH and the level of ROS were increased significantly (P<0.01), and the expressions of TNF-α and IL-1β protein were augmented in the heart of rats with MI (P<0.01). However, the expressions of SIRT1 mRNA and protein and the activity of SOD were obviously decreased in MI group (P<0.01). Furthermore, compared with the MI group, AIT increased the expressions of SIRT1 mRNA and protein and the activity of SOD in the heart of ME group (P<0.01); Meanwhile, the expressions of cardiac Nox4, MDA level, LDH activity and ROS level were diminished in ME group (P<0.01) as well as the decreased expressions of TNF-α and IL-1β protein (P<0.01). SIRT1 expression was negatively related to the expressions of NOX4 and ROS. Conclusion: AIT obviously inhibited myocardial oxidative stress and inflammatory reaction, improved cardiac function in rats with MI, and the mechanism was closely related to the activation of SIRT1-Nox4-ROS signaling pathway.
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A method of marking neurons on brain slices using patch clamp technique
GAO Jie, HUANG Yan-yun, ZHANG Yu-tong, DU Xiang-xin, ZHANG Li-na, HAO Na, GUO Xia, LI Jian-guo, ZHANG Yu
CJAP    2021, 37 (4): 445-448.   DOI: 10.12047/j.cjap.6035.2021.031
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Objective: To introduce a method of marking neurons using patch clamp technique. Methods: The brain slices of the target area was cut with a vibrating microtome. The glass microelectrode was perfused with the electrode liquid containing NeurobiotinTM Tracer, and the whole-cell patch-clamp recording was performed. After recording, the brain slices were fixed and rinsed with 4% paraformaldehyde. After stained in phosphate buffer with Streptavidin-Texas Red and Triton X-100 for at least 2 hours, the neurons can be observed under a fluorescence microscope. Results: The cell membrane voltage was clamped at -70 mV, and the neuron showed a gradually increasing membrane current after step stimulation. When recording in the current clamp mode, the step stimulus caused the neuron to depolarize to the threshold potential and then burst into action potentials. The morphology of intact neurons with clear cell body and protrusions of a neuron could be observed under a fluorescence microscope. Conclusion: This method is suitable for observing the morphological features of the recorded neuron after patch clamp experiments, which is easy to operate, and the image is intuitive and clear.
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Effect of different work rate increasing rate on the overall function evaluation of cardiopulmonary exercise testing I—peak parameters and changes in respiratory exchange rate
HAO Lu, SUN Xing-guo, SONG Ya, LIU Fang, TAI Wen-qi, GE Wan-gang, LI Hao, ZHANG Ye, CHEN Rong, , ZOU Yu-xin, , MA Ming-xin, , XIA Rui, , HUANG Yan, XIE You-hong
CJAP    2021, 37 (2): 113-119.   DOI: 10.12047/j.cjap.0083.2021.113
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Objective: To observe the effect of healthy volunteers different work rate increasing rate cardiopulmonary exercise test (CPET) on the peak exercise core indicators and the changes of respiratory exchange rate (RER) during exercise, to explore the effect of different work rate increasing rate on CPET peak exercise related indicators. Methods: Twelve healthy volunteers were randomly assigned to a moderate (30 W/min), a relatively low (10 W/min) and relatively high (60 W/min) three different work rate increasing rate CPET on different working days in a week. The main peak exercise core indicators of CPET data: VO2, VCO2, work rate (WR), breathe frequency(Bf), tidal volume (VT), ventilation (VE), heart rate (HR), blood pressure (BP), Oxygen pulse(O2P), exercise time and RER for each period of CPET were analyzed using standard methods. The ANOVA test and paired two-two comparison was performed on the difference of each index in the three groups of different work rate increasing rate. Results: Compared with the moderate work rate group, the peak work rate of the lower and higher work rate groups were relatively lower and higher, respectively ((162.04±41.59) W/min vs (132.92±34.55) W/min vs (197.42±46.14) W/min, P<0.01); exercise time was significantly prolonged and shortened ((5.69 ± 1.33) min vs (13.49 ± 3.43) min vs (3.56 ± 0.76) min, P<0.01); peak RER (1.27 ± 0.07 vs 1.18 ± 0.06 vs 1.33 ± 0.08, P<0.01~P<0.05) and the recovery RER maximum (1.72±0.16 vs 1.61±0.11 vs 1.81±0.14, P<0.01~P<0.05) were significantly decreased and increased. Conclusion: Different work rate increasing rate of CPET significantly change the Peak Work Rate, exercise time, Peak RER, and maximum RER during recovery. The CPET operator should choose an individualized work rate increasing rate that is appropriate for the subject, and also does not use a fixed RER value as a basis for ensuring safety, the subject's extreme exercise, and early termination of exercise.
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Effect of different work rate increasing rate on the overall function evaluation of cardiopulmonary exercise testing II— sub-peak parameters
HAO Lu, SUN Xing-guo, SONG Ya, LIU Fang, TAI Wen-qi, GE Wan-gang, LI Hao, ZHANG Ye, CHEN Rong, , ZOU Yu-xin, , MA Ming-xin, , XIA Rui, , HUANG Yan, XIE You-hong
CJAP    2021, 37 (2): 120-124.   DOI: 10.12047/j.cjap.0084.2021.114
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Objective: To observe the effect of healthy volunteers different work rate increasing rate cardiopulmonary exercise testing (CPET) on the sub-peak parameters . Methods: Twelve healthy volunteers were randomly assigned to a moderate (30 W/min), a relatively low (10 W/min) and relatively high (60 W/min) three different work rate increasing rate CPET on different working days in a week. The core indicators related to CPET sub-peak exercise of 12 volunteers were compared according to standard Methods: anaerobic threshold (AT), oxygen uptake per unit power (ΔVO2/ΔWR), oxygen uptake eficiency plateau,(OUEP), the lowest average of 90 s of carbon dioxide ventilation equivalent (Lowest VE/ VCO2), the slope of carbon dioxide ventilation equivalent (VE/ VCO2 Slope) and intercept and anaerobic threshold oxygen uptake ventilation efficiency value (VO2/ VE@AT) and the anaerobic threshold carbon dioxide ventilation equivalent value (VE/ VCO2@AT). Paired t test was performed on the difference of each parameter in the three groups of different work rate increasing rate. Results: Compared with the relatively low and relatively high work rate increasing rate group, the moderate work rate increasing rate group uptake eficiency plateau, (42.22±4.76 vs 39.54±3.30 vs 39.29±4.29) and the lowest average of 90 s of carbon dioxide ventilation equivalent (24.13±2.88 vs 25.60±2.08 vs 26.06±3.05) was significantly better, and the difference was statistically significant (P<0.05); Compared with the moderate work rate increasing rate group, the oxygen uptake per unit work rate of the relatively low and relatively high work rate increasing rate group increased and decreased significantly ((8.45±0.66 vs 10.04±0.58 vs 7.16±0.60) ml/(min·kg)), difference of which was statistically significant (P<0.05); the anaerobic threshold did not change significantly ((0.87±0.19 vs 0.87±0.19 vs 0.89±0.19) L/min), the difference was not statistically significant (P>0.05). Conclusion: Relatively low and relatively high power increase rate can significantly change the CPET sub-peak sports related indicators such as the effectiveness of oxygen uptake ventilation, the effectiveness of carbon dioxide exhaust ventilation, and the oxygen uptake per unit work rate. Compared with the moderate work rate increasing rate CPET, the lower and higher work rate increasing rate significantly reduces the effectiveness of oxygen uptake ventilation and the effectiveness of carbon dioxide exhaust ventilation in healthy individuals. The standardized operation of CPET requires the selection of a work rate increasing rate suitable for the subject, so that the CPET sub-peak related indicators can best reflect the true functional state of the subject.
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A preliminary report on the variation of respiratory heart rate during sleep in normal subjects and patients with chronic diseases without sleep apnea
WANG Ji-nan, SUN Xing-guo, XIE You-hong, SONG Ya, TAI Wen-qi, ZHOU Qing-qing, ZHANG Yan-fang, SHI Chao, XU Fan, LIU Fang, ZHANG Ye, HAO Lu, GE Wan-gang, LI Hao, XU Dan-dan
CJAP    2021, 37 (2): 125-134.   DOI: 10.12047/j.cjap.0102.2021.115
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Objective: The new theory of holistic integrative physiology and medicine, which describes the integrative regulation of respiratory, circulatory and metabolic systems in human body, generates the hypothesis of that breath is the origin of variability of circulatory parameters. We investigated the origin of heart rate variability by analyzing relationship between the breath and heart rate variability (HRV) during sleep. Methods: This retrospective study analyzed 8 normal subjects (NS) and 10 patients of chronic diseases without sleep apnea (CDs-no-SA). After signed the informed consent form, they performed cardiopulmonary exercise testing (CPET) in Fuwai Hospital and monitored polysomnography (PSG) and electrocardiogram (ECG) during sleep since 2014. We dominantly analyzed the correlation between the respiratory cycle during sleep and the heart rate variability cycle of the ECG R-R interval. The HRV cycle included the HR increase from the lowest to the highest and decrease from the highest to the lowest point. The number of HRV (HRV-n), average HRV time and other parameters were calculated. The breath cycle included complete inhalation and subsequent exhalation. The number of breath (B-n), average breath time and other breath parameters were analyzed and calculated. We analyzed each person's relationship between breath and HRV; and the similarities and differences between the NS and CDs-no-SA groups. Independent sample t test was used for statistical analysis, with P<0.05. Results: CPET core parameter such as Peak VO2 (83.8±8.9)% in NS were significantly higher than that (70.1±14.9)% in patients of chronic diseases without sleep apnea (P<0.05), but there was no difference between their AHI (1.7±1.3) in NS and AHI (2.9±1.2) in CDs-no-SA (P>0.05). The B-n and the HRV-n (6581.63±1411.90 vs 6638.38±1459.46), the average B time and the average HRV time (4.19±0.57)s vs (4.16±0.62)s in NS were similar without significant difference (P>0.05). The comparison of the numbers in CDs-no-SA were the number (7354.50±1443.50 vs 7291.20±1399.31) and the average times ((4.20±0.69)s vs (4.23±0.68)s) of B and HRV were similar without significant difference (P>0.05). The ratios of B-n/HRV-n in NS and CDs-no-SA were (0.993±0.027 vs 1.008±0.024) and both were close to 1 and similar without significant difference (P>0.05). The average magnitude of HRV in NS ((5.74±3.21) bpm) was significantly higher than that in CDs-no-SA ((2.88±1.44) bpm) (P<0.05). Conclusion: Regardless of the functional status of NS and CDs-no-SA, there is a similar consistency between B and HRV. The origin of initiating factors of HRV is the respiration.
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Preliminary analysis of the influence of breathing on heart rate variability in chronically ill patients with sleep apnea
ZHOU Qing-qing, SUN Xing-guo, WANG Ji-nan, TAI Wen-qi, SONG Ya, HAO Lu, ZHANG Ye, GE Wan-gang, LI Hao, ZHANG Yan-fang, SHI Chao, XU Fan, XU Dan-dan, XIE You-hong
CJAP    2021, 37 (2): 135-141.   DOI: 10.12047/j.cjap.0103.2021.116
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Objective: Based on the hypothesis that respiration causes variability of circulatory indicators proposed by the holistic integrated physiology and medicine theory, the correlation between respiration and heart rate variability during sleep in chronically ill patients with abnormal sleep breathing is analyzed. Methods: Eleven chronically ill patients with abnormal sleep breathing and apnea-hypopnea index (AHI) ≥15 times/hr are recruited. After signing the informed consent, they completed the standardized symptomatic restrictive extreme exercise cardiopulmonary exercise testing (CPET) and sleep breathing monitoring Calculate and analyze the rules of respiratory nasal airflow and ECG RR interval heart rate variability during the oscillatory breathing (OB) phase and the normal steady breathing phase of the patient during sleep, and use the independent sample t test to compare with normal people and no sleep breathing abnormalities in the same period in this laboratory. Of patients with chronic diseases are more similar and different. Results: The peak oxygen uptake and anaerobic threshold (AT) of CPET in chronic patients with abnormal sleep apnea were (70.8±13.6)% Pred and (71.2±6.1)% Pred; 5 cases of CPET had exercise induced oscillatory breathing (EIOB), 6 An example is unstable breathing, which indicates that the overall functional status is lower than normal. In this group of patients with chronic diseases, AHI (28.8±10.0) beats/h, the ratio of the total time of abnormal sleep breathing to the total time of sleep (0.38±0.25); the length of the OB cycle (51.1±14.4)s. The ratio (Bn/HRV-B-n) of the number of breathing cycles in the normal and steady breathing period to the number of heart rate variability cycles in this group of patients with chronic diseases is 1.00±0.04, and the CV (SD of HRV-B-M/x) is (0.33 ±0.11), blood oxygen saturation (SpO2) did not decrease significantly, the average amplitude of heart rate variability (HRV-B-M) of each respiratory cycle rhythm was (2.64±1.59) bpm, although it was lower than normal people (P<0.05) , But it was similar to chronic patients without sleep apnea (P>0.05). In this group of patients with chronic diseases, the ratio of the number of respiratory cycles to the number of heart rate variability cycles (OB-Bn/OB-HRV-B-n) during OB is (1.22±0.18), and the average amplitude of heart rate variability for each respiratory cycle rhythm in OB (OB -HRV-B-M) is (3.56±1.57)bpm and its variability (OB-CV = SD of OB-HRV-B-M/x) is (0.59±0.28), the average amplitude of heart rate variability in each OB cycle rhythm (OB-HRV-OB-M) is (13.75±4.25)bpm, SpO2 decreases significantly during hypoventilation during OB, and the average decrease in SpO2 during OB (OB-SpO2-OB-M) is (4.79±1.39)%. The OB-Bn/OB-HRV-B-n ratio, OB-HRV-OB-M and OB-SpO2-OB-M in the OB period are all significantly higher than the corresponding indicators in the normal stable breathing period Large (P<0.01). Although OB-HRV-B-M has no statistically significant difference compared with HRV-B-M in normal stable breathing period (P>0.05), its variability OB-CV is significantly increased (P<0.01). Conclusion: The heart rate variability of chronic patients with abnormal sleep breathing in the OB phase is greater than that of the normal stable breathing period. When the breathing pattern changes, the heart rate variability also changes significantly. The number of breathing cycles in the stable breathing period is equal to the number of heart rate variability cycles.The ratio is the same as that of normal people and chronically ill patients without sleep apnea, confirming that heart rate variability is respiratory origin; and the reduction of heart rate variability relative to the respiratory cycle during OB is directly caused by hypopnea or apnea at this time, and heart rate variability is also breathing source.
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Screening biomarkers for hypertensive heart disease: Analysis based on data from 7 medical institutions
ZHANG Xue-mei, ZHONG Xiao-gang, GONG Jun, TIAN Jun, ZHANG Yi, CHEN Ying-zhe, CUI Jing, WANG Zeng-zi, RAN Shu-qiong, XIANG Tian-yu, XIE You-hong, SUN Xing-guo
CJAP    2021, 37 (2): 142-146.   DOI: 10.12047/j.cjap.0093.2021.117
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Objective: To screen the influencing factors of hypertensive heart disease (HHD), establish the predictive model of HHD, and provide early warning for the occurrence of HHD. Methods: Select the patients diagnosed as hypertensive heart disease or hypertensionfrom January 1, 2016 to December 31, 2019, in the medical data science academy of a medical school. Influencing factors were screened through single factor and multi-factor analysis, and R software was used to construct the logistics model, random forest (RF) model and extreme gradient boosting (XGBoost) model. Results: Univariate analysis screened 60 difference indicators, and multifactor analysis screened 18 difference indicators (P<0.05). The area under the curve (AUC) of Logistics model, RF model and XGBoost model are 0.979, 0.983 and 0.990, respectively. Conclusion: The results of the three HHD prediction models established in this paper are stable, and the XGBoost prediction model has a good diagnostic effect on the occurrence of HHD.
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Max test verify further clinical research for whether individualized symptom-limited cardiopulmonary exercise testing is the maximum extreme exercise
ZHANG Ye, SUN Xing-guo, LIU Fang, HAO Lu, SONG Ya, TAI Wen-qi, GE Wan-gang, LI Hao, SHI Chao, XU Fan, WANG Ji-nan, ZHANG Yan-fang, ZHOU Qing-qing, CHEN Rong, , ZOU Yu-xin, MA Ming-xin, , XIA Rui, HUANG Yan, XIE You-hong
CJAP    2021, 37 (2): 147-153.   DOI: 10.12047/j.cjap.0080.2021.118
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Objective: To verify that the cardiopulmonary exercise testing (CPET) performed by clinical subjects is the maximum extreme exercise, we designed The Max test(Max)during clinical CPET. We used Max to verify the accuracy of the quantitative CPET evaluation result, and whether it is feasible and safe to use the specific value of a certain index as the standard for stopping CPET. Methods: Two hundred and sixteen cases from Fuwai Hospital were selected during June 2017 to January 2019,including 41 healthy person(control group) and 175with cardiovascular diseases(patient group),The patients had a CPET peak RER ≤ 1.10, or the peak heart rate and peak blood pressure were basically non-responsive.The Max was first attempted in 60 subjects,and this study is further expanded . When the CPET ended, they had a 5-minute break, then the Max, during which, they cycled with a velocity of ≥ 60 r/min, at a constant intensity equivalent to to 130% of peak work,until exhausted.The difference and percentage difference between the peak heart rate and the peak oxygen uptake were calculated. ①If the percentage difference of heart rate and oxygen uptake are all less than -10%,then the Max is defined as failure,otherwise it is succesful. 2 If the percentage difference is between -10%~10%, then the Max is successful, which proved that the CPET is precise.③If the difference is ≥10%, the Max is successful, which proves that the CPET is non-extreme exercise. Results: Patient group's Peak VO2(L/min,ml/(min·kg)),anaerobic threshold (L/min,ml/(min·kg),%pred),Peak VO2/HR(ml/beat, % pred),Peak RER,Peak SBP,Peak WR,peak heart rate,OUEP (ratio,%pred) were lower than those of the control group(P<0.05).The VE/ VCO2 Slope (ratio,%pred)and Lowest VE/ VCO2(ratio,%pred) were higher in the patient group than in the control group (P<0.05).No adverse events occurred during the CPET and Max in all cases. Among the 216 cases,Max was successful in 198 cases(91.7%).CPET was proved to be maximum extreme exercise for 182 cases,non-maximum extreme exercise for 16 cases,and failed in 18 cases(8.3%).Conclusion: For CPET with a low peak RER and a maximum challenge,the Max can confirm the accuracy of the objective quantitative assessment of CPET. Max is safe and feasible,and that deserved further research and clinical application.
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A clinical research report on the pathophysiological characteristics of exercise in patients with mitral regurgitation
Mu xiya, SUN Xing-guo, TAI Wen-qi, WANG Ji-nan, ZHOU Qing-qing, ZHANG Yan-fang, SONG Ya, SHI Chao, XU Fan, XU Dan-dan, SUN Shu-yan
CJAP    2021, 37 (2): 154-161.   DOI: 10.12047/j.cjap.0105.2021.119
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Objective: Cardiopulmonary exercise testing (CPET) was used to investigate the exercise pathophysiology of mitral regurgitation. Methods: 26 patients with moderate and severe mitral regurgitation who completed standardized extreme exercise CPET under strict quality control after signing informed consent since 2016, and 11 normal subjects in the same period as the control group. The core indexes of CPET were analyzed and calculated according to the standard method and compared with normal subjects for intergroup statistical independent sample t-test. At the same time, the patients with heart failure and exercise oscillation breathing (OB) were divided into two subgroups: 11 cases without heart failure, 15 cases with heart failure, 8 cases with non-OB and 18 cases with OB, and their similarities and differences were compared between each subgroup. Results: The core indexes of CPET, such as peak oxygen uptake (85.60 ±9.06)%pred and anaerobic threshold (AT, (87.59 ±15.38)%pred) were normal. The peak oxygen uptake of CPET in patients with mitral regurgitation was (48.15 ±12.11)%pred, peak oxygen pulse was (66.57 ±12.20)%pred, AT was (56.75 ±11.50)%pred, oxygen uptake efficiency plateau was (88.24 ±16.42)%pred , lowest value of carbon dioxide ventilatory efficiency was (125.89 ±27.05)%pred and slope of carbon dioxide ventilatory efficiency was (128.31 ±31.68)%pred. Among them, only oxygen uptake efficiency plateau (OUEP) was normal and low, and the other indexes were significantly abnormal. There were significant differences between the patients and the control group (P<0.01). There was no significant difference between the non-OB group and the OB group, but there was significant difference between the non-OB group and the control group (P<0.05). There was no significant difference between the non-heart failure group and the heart failure group, but there was significant difference between the non-heart failure group and the control group. Conclusion: All the core indexes of cardiopulmonary exercise are significantly abnormal in patients with mitral regurgitation who are significantly lower than those in normal subjects except for the low effectiveness of oxygen ventilation. And with or without heart failure and OB did not affect the cardiopulmonary function.
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Clinical study on the characteristics of exercise pathophysiological in patients with severe heart failure
CHEN Rong, SUN Xing-guo, ZHANG Ye, ZOU Yu-xin, FENG Yun-hong, MA Ming-xin, XIA Rui, WANG Dong, HUANG Yan, LI Hao, YANG Ge
CJAP    2021, 37 (2): 162-168.   DOI: 10.12047/j.cjap.0074.2021.120
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Objective: The cardiopulmonary function of patients with chronic heart failure (CHF) was severely limited, but the holistic integrative exercise pathophysiology is still unclear. Methods: After signed the consent form, Eighty three patients with severe CHF from October 2016 to October 2017 in Fuwai Hospital were performed Ramp incremental loading program CardioPulmonary Exercise Testing (CPET), and 12 normal subjects served as control. CPET were performed according to standard of Harbor-UCLA MC and the circulatory, respiratory and metabolic parameters during CPET were measured and analyzed. Results: Peak oxygen uptake (Peak VO2) in CHF (14.33±2.69) ml/(min·kg), (44.25±14.74)%pred was significantly lower than control ((29.42±5.46) ml/(min·kg), (83.88±6.28)%pred). Other core parameters of CPET such as anaerobic threshold (AT), peak oxygen pulse, oxygen uptake efficiency platform (OUEP), the lowest of carbon dioxide output ventilation ratio (Lowest VE/VCO2), and carbon dioxide output ventilation slope (VE/VCO2 Slope) in CHF were significantly different with the control group(P<0.01). The core parameters of lung function, such as forced expiratory volume in the first second (FEV1), forced vital capacity (FVC), FEV1/FVC, and carbon monoxide diffusion (DLCO) were significantly decreased (P<0.01). Systolic blood pressure during all stages of CPET in CHF was significantly lower than control group (P<0.05); Heart rate at AT, peak and recovery stages were significantly lower than control (P<0.01). Minute ventilation, tidal volume and respiratory frequency at rest, warm-up were significantly higher than control (P<0.05). Tidal volume at recovery was significantly higher than control (P<0.05). VO2 at AT, peak and recovery stages in CHF were significantly higher than control (P<0.01). Oxygen pulse at AT and peak were significantly higher than control (P<0.01). Pulse oxygen saturation during all stages of CPET in CHF were significantly lower than control (P<0.01). Conclusion: The decreased holistic functional capacity of cardiogenic CHF dominantly due to circulatory limitation, and secondly due to respiratory and metabolic limitation.
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