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  • Table of Content
      28 July 2018, Volume 34 Issue 4 Previous Issue    Next Issue
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    Protective effects of exogenous H2S to the recovery of hypoxia post-conditioning on aged H9C2 cells and the underling mechanisms
    SUN Wei-ming, ZHANG Yuan-zhou, WEN Xin, XI Yu-xin, YUAN Di, WANG Yue-hong, WEI Can, XU Chang-qing, LI Hong-zhu
    CJAP. 2018, 34 (4): 289-293.   DOI: 10.12047/j.cjap.5642.2018.067
    Abstract   PDF (1510KB) ( 134 )
    Objective: To investigate the recovery of protective effects of exogenous hydrogen sulfide (H2S) on hypoxia post-conditioning in aged H9C2 cells and its mechanism. Methods: H9C2 cells (cardiomyocytes line) were treated with 30 μmol/L hydrogen peroxide (H2O2) for 2 hours, then cultured for 3 days in order to induce cellular aging. Aged H9C2 cells were randomly divided into 5 groups (n=8):Control group (Control), hypoxia/reoxygenation group (H/R), H/R + NaHS group, hypoxia post-conditioning (PC) group, PC+NaHS group. H/R model:the cells were exposed to hypoxic culture medium (serum and sugar free medium, pH=6.8) for 3 hours and then cultured at normal condition for 6 hours. PC model:at the end of hypoxia for 3 hours, the cells were exposed to normoxic culture solution for 5 minutes, then the cells were placed in hypoxic solution for 5 minutes, the cycle above-mentioned was repeated 3 times and followed by reoxygenation for 6 hours. Advanced glycation end products (AGEs) content and caspase-3 activity were detected by ELISA. The cell viability was observed by cell counting kit-8 (CCK-8). The reactive oxygen species (ROS) levels were analyzed using 2, 7-dichlorodihydrofluorescein diacetate (DCFH-DA) staining. The apoptotic rate was determined through Hoechst 33342 staining. The mRNA levels of relative gene expression were detected by real-time PCR. Results: Thirty μmol/L H2O2 induced H9C2 cell senescence while did not lead to apoptosis. Compared with control group, cell viability was decreased, the apoptotic rate、levels of ROS and the mRNA of caspase-3, caspase-9 and Bcl-2 were increased in H/R and PC groups (P<0.01). There were no differences in the above indexes between PC group and H/R group. Supplementation of NaHS increased cell viability and decreased apoptotic rate and oxidative stress. The effects of PC + NaHS on the above indexes were better than those of H/R+NaHS group. Conclusion: Exogenous H2S can restore the protective effect of PC on the aged H9C2 cells, and its mechanism is related to the inhibition of oxidative stress and apoptosis.
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    Effect of EPO on PRDM16, FGF21 expression and STAT phosphorylation of brown adipose tissue in HFD mice
    ZHANG Rui, CHEN Lu, GE Jun-mei, MA Gen-shan, CAI Jun-yan
    CJAP. 2018, 34 (4): 294-298.   DOI: 10.12047/j.cjap.5654.2018.068
    Abstract   PDF (1392KB) ( 250 )
    Objective: To investigate the effect of erythropoietin (EPO) on blood glucoseand plasma insulin level, index of insulin resistance (HOMA-IR), introperitoneal glucose tolerance test (IPGTT), the mRNA and protein level of PR domain-containing 16 (PRDM16), phosphorylated signal transducer and activator of transcription 3 (p-STAT3), fibroblast growth factor 21 (FGF21) of brown adipose tissue (BAT) in mice fed with high fat diet (HFD) in order to provide clues for the mechanism of obesity and complication.Methods: Twenty C57BL/6J male mice fed with HFD were randomly divided into control group (HFD-Con) and EPO group (HFD-EPO), mice in the two groups were injected intraperitoneally normal saline and EPO (200 IU/kg) res pectively, 3 times per week for consecutive 4 weeks.Then the body weight, blood glucose, plasma insulin level, HOMA-IR and IPGTT were detected.The mRNA and protein level of PRDM16, FGF21, p-STAT3/STAT3 in brown adipose tissue were detected by real-time quantitative RT-PCR and Western blot respectively.Results: After intraperitoneal injection of EPO for 4 weeks, the body weight of the mice in HFD-EPO and HFD-Con groups was (26.65±0.85) g and (31.50±1.6 0) g respectively.The blood glucose of the mice in HFD-EPO group[(62.79±8.09) mg/dl]was significantly decreased compared with that in HFD-Con group[(91.06±9.86) mg/dl].The plasmainsulin level in HFD-EPO group[(10.56±1.06)μU/ml]was significantly decreased compared with that in HFD-Con group[(13.2±1.1)μU/ml, P< 0.01].The level of IPGTT in HFD-EPO group was significantly ameliorated and th e HOMA-IR decreased compared with those in HFD-Con group.The mRNA and protein expressions of PRDM16, FGF21 and the level of STAT3 of brown adipose tissue in HFD-E PO group were increased obviously.And there was no difference of FGF21 mRNA content in liver and FGF21 content in plasmabetween the two groups.Conclusion: EPO could promote differentiation of brown adipose tissue by increase in the express ion of PRDM16, and decrease the blood glucose level, ameliorate glucose metabolism in obses mice.
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    Effect of particulate matter 2.5 at urban centre of Hangzhou on lung impairment in rats
    TIAN Cheng-yuan, YAN Ming, ZHANG yun, LIU Ke-zhou, GUO Miao, SUN Yong-hong, XUE Ling-yun, ZHU Lei, XU Ying
    CJAP. 2018, 34 (4): 299-303.   DOI: 10.12047/j.cjap.5651.2018.069
    Abstract   PDF (1254KB) ( 148 )
    Objective: To explore the effect of ambient particle matter 2.5 (PM2.5) collected in the urban center of Hangzhou on the lung injury of rats and on the activating of endoplasmic reticulum pathway. Methods: PM2.5 samples were collected on quartz fiber filters using a PM2.5 high-volume air sampler in the urban area of Hangzhou. The collected PM2.5 particles were extracted in ultrapure water and concentrated by vacuum freeze-drying. Twenty-four male Sprague-Dawly (SD) rats were randomly divided into 3 groups:saline control group, low dose PM2.5 exposure group (5 mg/kg BW) and high dose PM2.5 exposure groups (25 mg/kg BW). Each group received intratracheal instillation of PM2.5, once a week for 4 weeks. Twenty-four hours after the last exposure, the rats were narcotized and sacrificed, left lung was isolated and fixed with 4% paraformaldehyde for histopathological detection. The bronchoalveolar lavage fluid (BALF) was collected from the right lung. The total antioxidant capacity (T-AOC) level, the activities of superoxide dismutase (SOD) and lactic dehydrogenase (LDH) in BALF were detected by chemical colorimetry. The level of tumor necrosis factor-alpha (TNF-α), interleukin-1 beta (IL-1β) and interleukin-6 (IL-6) cytokines in BALF was measured by enzyme linked immunosorbent assay (ELISA). And the protein expressions of glucose-regulated protein 78 (GRP78), phosphorylated protein kinase receptor-like endoplasmic reticulum kinase (p-PERK), phosphorylated eukaryotic translation initiation factor (p-eIF2α), transcription factors C/EBP homologue protein (CHOP), inositol-requiring enzyme 1α (IRE1α) and X-box binding protein 1 (XBP1) in lung tissue were determined by Western blotting. Results: Compared with control group, rats in both low dose (5 mg/kg) and high dose (25 mg/kg) PM2.5-treated groups showed obviously dose-dependent pulmonary toxicity including thickening of alveolar walls, narrowing of alveolar space, interstitial hyperplasia and inflammatory cell infiltration. Compared with control group, T-AOC level and the SOD activity in BALF in both PM2.5-treated groups were decreased dose-dependently (P<0.05), whereas the LDH activity in BALF were increased in a dose-dependent manner (P<0.05). Exposure to PM2.5 resulted in a increasing of the release of proinflammatory cytokines including TNF-α, IL-1β and IL-6 in rat lung in a dose-dependent manner (P<0.05). The levels of GRP78, p-PERK, p-eIF2α, CHOP, IRE1α and spliced XBP1 (XBP1-S) were significantly up-regulated, whereas the level of unspliced XBP1 (XBP1-U) was down-regulated in the rat lung tissue of high-dose PM2.5 treated group. Conclusion: The PM2.5 in the urban area of Hangzhou can significantly cause lung inflammatory injury in rats. Both oxidative stress and activation of ER stress pathways may be related to such PM2.5 inhalation-induced lung inflammatory injury.
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    Effects of miRNA-1,miRNA-21 in plasma on in-stent restenosis in patients with coronary heart disease and diabetes mellitus after percutaneous coronary intervention
    GUAN Jing-jing, ZHANG Ying, LIU Yu-jie
    CJAP. 2018, 34 (4): 304-308.   DOI: 10.12047/j.cjap.5643.2018.070
    Abstract   PDF (1219KB) ( 177 )
    Objective: To observe the expression differences of the plasma miRNA-1, miRNA-21 between patients with coronary heart disease (CHD) and without coronary artery lesions, between patients with in-stent restenosis (ISR) and none in-stent restenosis (NISR), and to study their predictive value for ISR occurred after percutaneous coronary intervention (PCI) in patients with CHD and diabetes mellitus (DM). Methods: The selected subjects were divided into CHD group in which patients were implemented stenting (n=187), and control group in which patients were without coronary artery lesions (n=195). According to the guidelines, the control group was divided into normal group (n=150), simple-DM group (n=45); the CHD group was divided into simple-CHD group (n=119) and CHD-DM group (n=68), the CHD group was also divided into ISR group (n=48), NISR group (n=139), and the ISR group was divided into simple-ISR group (n=26) and ISR-DM group (n=22) again. Plasma was collected from each group, and total RNA was extracted, the level of blood miRNA-1, miRNA-21 of each group was detected, and their level differences were analyzed. Results: Compared with control group, the level of miRNA-1 and miRNA-21 of CHD group was increased (P<0.05); compared with NISR group, the level of miRNA-1 and miRNA-21 of ISR group was increased (P<0.05). The incidence of ISR of CHD-DM group was obviously higher than that of simple-CHD group, ISR-DM group's level of miRNA-21 was higher than that of simple-ISR group (P<0.05), and there was no difference of miRNA-1 level between ISR and ISR-DM group (P<0.05). In Logistics, for CHD patents, the OR of DM, miRNA-1, miRNA-21 were 2.132, 3.066, 1.924 respectively (P<0.05); for CHD patents with ISR, the OR of DM, miRNA-21 were 2.123, 3.066 respectively (P<0.05); especially for CHD and DM patents with ISR, the OR of miRNA-21 was 9.148 (P<0.05). In ROC curve, for CHD patients with ISR, the AUC of miRNA-1, miRNA-21 were 0.854, 0.857 respectively; for CHD-DM patients with ISR, the AUC of miRNA-21 was 0.783. Conclusion: To predict the occurrence of ISR for CHD patients, the plasma miRNA-1 and miRNA-21 have a relatively high specificity and sensitivity, for CHD patients with DM, miRNA-21 may have a higher clinical value.
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    Effect of ursolic acid on cardiomyopathy of mice with diabetes and its mechanism
    YANG Zhang-liang, XU Hui-lin, CHENG Yin, ZHAO Jin-guo, ZHOU Yu-jie, WENG Yang-jing, WANG Xu-tao, QI Min-you
    CJAP. 2018, 34 (4): 309-312.   DOI: 10.12047/j.cjap.5638.2018.071
    Abstract   PDF (1183KB) ( 241 )
    Objective: To study the effect of ursolic acid on cardiomyopathy in mice with diabetes induced by high-fat diet combined with low dose streptozotocin, and to explore its possible mechanism. Methods: Thirty male ICR mice were randomly divided into control group (n=10) and moulding group (n=20), the mice in the two groups were fed with regular diet and high-fat diet respectively for 6 weeks, and then the mice in the moulding group were injected with streptozotocin (30 mg/kg) for 5 successive days to induce diabetes mellitus (DM). Fasting blood glucose (FBG) was measured after 9 days. Mice with FBG over 11.1 mmol/L were regarded as DM. Twenty DM mice were randomly divided into model group and ursolic acid group (n=10). Mice in each group were continuously administrated ursolic acid (100 mg/kg) or corresponding solvent intragastrically for 8 weeks. After that, FBG was measured, body weight (BW), heart weight and left ventricular weight were weighed in order to calculate the heart mass index (HMI) and left ventricular mass index (LVMI). Levels of creatine kinase (CK), lactate dehydrogenase (LDH) in serum and the level of superoxide dismutase (SOD), malondialdehyde (MDA) in myocardial tissue were detected. HE staining was used to observe pathological changes of myocardial tissue. Immunohistochemistry was employed to determine the expression of NOD-like receptor protein 3 (NLRP3) and interleukin 1β (IL-1β). Results: Compared with the control group, HMI, LVMI were apparently enlarged, levels of FBG, CK, LDH in serum and MDA in myocardial tissue were extremely increased, while the activity of SOD in myocardial tissue were extraordinary decreased in diabetic group. HE staining of myocardium showed that arrangement disorder of myocardial fibers, edema and hypertrophy in myocardial cell, as well as inflammatory cell infiltration in model group. Immunohistochemistry showed that the expression of NLRP3 and IL-1β in myocardial tissue increased obviously in model group, the above changes inursolic acid group were significantly ameliorated. Conclusion: Ursolic acid has a obvious protective effect on myocardial injury in mice with diabetes induced by high-fat diet combined with low dose streptozotocin, and its mechanism may be associated with inhibiting NLRP3 inflammasome activation, reducing IL-1β generation and alleviating myocardial inflammatory injury.
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    Protective effect of simvastatin on kidney of rats with diabetes mellitus and the possible mechanism
    WAN Xin, WANG Xi, LI Fan-lu, WU Ya-li, LIU Xin, CHEN Huan-zhen, YANG Jing, CUI Xiang-li
    CJAP. 2018, 34 (4): 313-317.   DOI: 10.12047/j.cjap.5636.2018.072
    Abstract   PDF (1178KB) ( 133 )
    Objective: To observe the protective effect of simvastatin on renal injury in diabetic rats and to explore the possible molecular mechanism. Methods: Twenty-four SD rats were randomly divided into normal control (NC) group (n=8) and modeling group (n=16).The rats in modeling group were injected with streptozotocin intraperitoneally at a dose of 55 mg/kg to establishing diabetic rat model. After diabetic ratmodel established successfully, the diabetic rats were randomly subdivided into diabetes mellitus (DM) group and diabetes mellitus + simvastatin (DM+Sim) group (n=8).Rats in DM+Sim group were given simvastatin at a dose of 40 mg/kg by oral gavages, once a day for 4 weeks. Morphological changes and interstitial fibrosis of kidney were observed by histopathological method. The expressions of relative protein in endoplasmic reticulum stress, inflammatory molecules in renal tissues and cells apoptosis were detected by molecular biology method. Results: ① Compared with NC group, the pathological changes of glomerulus and tubulointerstitium were obvious, and the collagen fibers were obviously erythrophilous and unevenly distributed in DM group. Compared with DM group, the morphological changes and fibrosis were significantly improved in DM+Sim group. ② The expressions of GRP78, p-IRE1α, NF-κB p65 and MCP-1 in DM group were significantly higher than those in NC group (P<0.05), while the expressions of GRP78, p-IRE1α, NF-κB p65 and MCP-1in DM + Sim group were decreased (P<0.05). ③ There were a small number of apoptotic nuclei in the glomeruli and adjunctive renal tubules in NC group detected by TUNEL assay, while there were a large number of apoptotic nuclei in DM group (P<0.01). The number of apoptotic nuclei was decreased significantly in DM+Sim group (P<0.01). Conclusion: Morphologicalchanges and fibrosis of renal tissue are improved obviously, and the number of apoptotic cells is decreased significantly after administration of simvastatin in diabetic rats. Simvastatin exertsthe protective effect on diabetic nephropathy by inhibiting endoplasmic reticulum stress and NF-κB inflammatory signaling pathway, and reducing renal cell apoptosis.
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    The relationship between autophagy activation in spinal cord and type 2 diabetic neuropathic pain in rats
    ZHU Ya-bing, JIA Gai-li, LU Jia-hui, ZHANG Mao-biao, LI Jun, CAO Hong
    CJAP. 2018, 34 (4): 318-323.   DOI: 10.12047/j.cjap.5644.2018.073
    Abstract   PDF (1349KB) ( 268 )
    Objective: To investigate the relationship between autophagy function in spinal cord and type 2 diabetic neuropathic pain in rats. Methods: Forty-two male Sprague-Dawley rats were fed with a high-sugar, high-fat diet for 8 weeks to induce the insulin resistance, and then received a single intraperitoneal streptozocin (STZ) injection to establish type 2 diabetes rat model. Two weeks after STZ injection, mechanical withdrawal threshold (MWT) and thermal withdrawal latency (TWL) of rats were detected, the rats with MWT and TWL decreasing to below 80% compared to baseline were chosen as type 2 diabetic neuropathic pain rats (group DNP, n=24), the rest of the rats were chosen as type 2 diabetic non-neuropathic pain rats (group DA, n=18). And another 18 normal rats randomly selected from the total were classified as control group (group C) and fed with common forage for 8 weeks. The MWT and TWL were measured again on the 3rd, 7th and 14th day after determining the grouping of DA and DNP, and then, the lumbar segments 4~6 of the spinal cord were removed from the executed rats for determination of the expressions of microtubule-associated protein light chain 3 (LC3)、Beclin-1and P62 by Western blot. The co-expressions of P62 with GFAP or OX-42 or NeuN in spinal dorsal horn were detected in another 6 lumbar segments of diabetic neuropathic pain (DNP) rats on the 7th day by immunofluorescence double dye method. Results: Compared with group C, the insulin level was increased and ISI decreased in SD rats fed with high-sugar, high-fat diet, that meant the rats in insulin-resistance. After STZ injection, blood glucose rose to the standard of type 2 diabetes mellitus, i.e. ≥ 16.7 mmol/L. Compared with group C and group DA, MWT was significantly decreased, TWL shortened and the expression of LC3-Ⅱ and Beclin-1 in the spinal dorsal horn up-regulated, P62 expression down-regulated on the 3rd, 7th and 14th day in group DNP (P<0.05). P62 was mainly localized in spinal dorsal horn and coexisted with neurons, and spots of P62 immunoreactivity could be detected in a few microglia but not observed in astrocyte. Conclusion: The changes in expression of LC3-Ⅱ、Beclin-1 and P62 in spinal cord of type 2 diabetes neuropathic pain rats means autophagy activation of spinal, up-regulated autophagy of neurons in spinal dorsal horn mainly involves in the formation and development of type 2 diabetic neuropathic pain in rats.
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    The role of endoplasmic reticulum stress in pulmonary hypertension in rat induced by chronic hypoxia and hypercapnia
    ZHANG Jing-jing, CHEN Jun-hao, ZHAO Mei-ping, WU Yuan-ling, ZHANG Cong-cong, YING Lei, CHEN Xi-wen, WANG Wan-tie
    CJAP. 2018, 34 (4): 327-333.   DOI: 10.12047/j.cjap.5644.2018.075
    Abstract   PDF (1421KB) ( 170 )
    Objective: To observe the pulmonary vascular remodeling in rats with pulmonary hypertension induced by hypoxia and hypercapnia, and to explore the role of endoplasmic reticulum stress in pulmonary hypertension.Methods: Forty SD rats were random-ly divided into four groups:normoxic control group (N), hypoxia hypercapnia group (HH), ERS inhibitor 4-phenylbutyric acid group (4-PBA), endoplasmic reticulum stress (ERS) pathway agonist tunicamycin group (TM), ten rats in each group.The mean pulmona-ry artery pressure (mPAP), mean carotid artery pressure (mCAP) and right ventricular hypertrophy index of rats in each group were measured.Pulmonary artery smooth muscle cells were identified by immunofluorescence α-smooth muscle actin (α-SMA).Morphologi-cal changes of lung tissue and pulmonary artery were observed by electron microscope.The apoptotic index of pulmonary artery smooth muscle cells in each group was detected by TUNEL.Reverse transcription polymerase chain reaction (RT-PCR) and Western blot were used to detect the expression of glucose-regulated protein (GRP78), C/EBP homologous protein (CHOP), c-Jun N-terminal kinase (JNK) and cysteinyl aspartate specific proteinase-12 (caspase-12) mRNA and protein in each group.Results: ①Compared with the N group, the mPAP, the ratio of right ventricle weight to left ventricle plus ventricular septum weight[RV/(LV+S)]and the ratio of pulmonary artery wall area to total tube area (WA/TA) were increased (P<0.01), and the ratio of pulmonary artery luminal area to total tube area (LA/TA) were decreased (P<0.01), pulmonary artery smooth muscle cell apoptosis index were decreased (P<0.05 or P<0.01) in HH group, 4-PBA group and TM group.ERS related protein and mRNA expressions were increased, the differences were statistically significant.②Compared with the HH group, the mPAP, [RV/(LV+S)]and WA/TA of 4-PBA group were decreased (P <0.01), LA/TA and pulmonary artery smooth muscle cell apoptosis index were increased (P<0.01, P<0.05).The expressions of ERS related protein and mRNA were all decreased (P<0.05 or P<0.01).③Compared with the HH group, the mPAP, [RV/(LV+S)]and WA/TA of TM group were increased (P<0.05 or P<0.01), pulmonary artery middle layer thickened, LA/TA and pulmonary artery smooth muscle cell apoptotic index were decreased (P<0.01).ERS related protein and mRNA expressions were increased with statistical significance except GRP78 protein.Conclusion: Pulmonary vascular remodeling in rats with pulmonary hypertension induced by hypoxia and hypercapnia may be related to the excessive proliferation of pulmonary artery smooth muscle cells and too little apopto-sis;ERS related factors (JNK, caspase-12 and CHOP) are involved in the regulation of pulmonary hypertension induced by hypoxia hypercapnia.
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    Prevalence and risk factors of chronic mountain sickness in Pamirs plateau
    YANG Sheng-hong, LI Bin, GAO Liang, LI Nian-hua
    CJAP. 2018, 34 (4): 336-339.   DOI: 10.12047/j.cjap.5607.2018.077
    Abstract   PDF (925KB) ( 41 )
    Objective: To investigate the prevalence of chronic mountain sickness (CMS) and its predisposing factors in Pamirs plateau and analysis clinical feature and risk factors. Methods: Eight hundred and twenty-four individuals in Pamirs plateau were surveyed used Qinghai CMS scoring system. Demographics, BP, SaO2, heart rate and specified symptoms of CMS were recorded, Hb level was estimated. Results: Overall CMS prevalence was 22.08%, including 21.84% mild and 0.24% moderate without severe patient. Gender, nation, age, immigration time, altitude, occupation and accommodation mode were risk factors for CMS (P<0.05 or P<0.01), CMS prevalence was higher than HAPC rate both in male and female (χ2=90.59 and 44.13, P<0.01). Multiple linear regression analysis showed age, BMI, SaO2 and systolic pressure correlated with CMS (P<0.01). Conclusion: Chronic mountain sickness prevalence in Pamirs plateau is rather high but high altitude polycythemia prevalence is low, so the disease type is high altitude deterioration. Work and exercise more reasonable to avoid overtiredness, oxygen inhalation to raise blood oxygen saturation, control blood pressure and resting therapy can improve CMS symptom and decrease prevalence.
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    Comparison of differential metabolites in urine of the middle school students with chronic fatigue syndrome before and after exercise
    CHI Ai-ping, WANG Zi-nan, SHI Bing, YANG Xiao-fan, MIN Rui-xin, SONG Jing
    CJAP. 2018, 34 (4): 340-344.   DOI: 10.12047/j.cjap.5633.2018.078
    Abstract   PDF (1391KB) ( 184 )
    Objective: To study the differential metabolites in urine and the characteristics of metabolic pathway of middle school students with chronic fatigue syndrome (CFS) before and after exercise, and then explain the metabolic mechanism of CFS. Methods: Eight male middle school students (age:17-19) with CFS were selectedas the CFS group according to CFS screening criteria of the U.S. centers.At the same time, 8 male health students of the same age from the same school were selected as the control group. They were administrated to do one-time exercise on the improved Harvard step (up and down steps 30 times/min for 3minutes). Their urinewascollected before and after exercise, and the differential metabolitesin urine were detected by liquid chromatography-mass spectrometry (LC-MS). The multidimensional statistical methods were used to analyze the metabolites by principal component analysis (PCA) and orthogonal projections to latent structures-discriminant analysis (OPLS-DA). Finally, MetPA database was used to analyze the metabolites and to construct the correlativemetabolic pathways. Results: Compared with the control group, the creatine, indoleacetaldehyde, phytosphingosine and pyroglutamic acid were selected as differential metabolites and the contents of those were decreased significantly (P<0.05 or P<0.01) in CFS groupbefore the step movement. However, 11 differential metabolitesin CFS group were selected out after exercise, which were nonanedioic acid, methyladenosine, acetylcarnitine, capric acid, corticosterone, creatine, levonorgestrel, pantothenic acid, pyroglutamic acid, xanthosine and xanthurenic acid in sequence, the contents of methyladenosine and creatinewere significantly increased (P<0.05) and the contents of the other 9 differentialmetabolites were significantly decreased (P<0.05 or P<0.01)compared with the control group.The 15 differential metabolites mentioned above were input MetPA database in order to analyze the metabolic pathways weighted score.The results showed that the arginine-proline metabolism pathway disordersweredetected in theCFS group before exercise, the marker metabolite wascreatine. And 3 metabolic pathwaysdisorder weredetectedin the CFS groupafter exercise, which were arginine-proline metabolism, biosynthesis of pantothenic acid and CoA, steroid hormone biosynthesis, and the marker metabolites, in turn, werecreatine, pantothenic acid and corticosterone. Conclusion: The disorder of arginine-proline metabolic pathway is detected in CFS middle school students before exercise intervention. After exercise, it can be detected that the steroid hormone biosynthetic metabolic pathway, pantothenic acid and CoA metabolic pathways also have metabolic disorders.
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    Effect of exercise on adiponectin in aged obese rats
    SU Min, BAI You-ping, SONG Wei-wei, WANG Ming, SHEN Rui-rui, DU Kang-jian, XIA Xing-quan, NIE Liu-wang
    CJAP. 2018, 34 (4): 345-349.   DOI: 10.12047/j.cjap.5660.2018.079
    Abstract   PDF (984KB) ( 106 )
    Objective: To explore effects of exercise on the expression of adiponectin mRNA and protein in visceral adipose tissue, plasma adiponectin concentration, and insulin resistance of aged obese rats. Methods: Male SD rats age to 21 days old were fed with high-fat diet (fat percentage was 36.3% to 40.0%) for three stages of adolescence, maturity and old age to establish elderly obesity rats model. When the rats aged to 60 weeks old, natural growing elderly rats were randomly divided into control group (C) and aged exercise group (AE), n=6; elderly obesity rats were randomly divided into obesity control group (OC) and obesity exercise group (OE), n=6. The treadmill grade was 0°, the exercise speed and time were 15 m/min×15 min, 4 groups each time, between consecutive groups the rats had 5 minutes rest, the rats were exercised for 60 minutes every day, five days a week, continuous exercise for 8 weeks. Then, the expressions of adiponectin mRNA and protein in visceral adipose tissue were determined. The concentrations of blood glucose, plasma adiponectin and insulin were measured. Insulin resistance was calculated. Results: Comparison with control group, the expressions of adiponectin mRNA and protein were obviously decreased, the concentration of blood glucose and insulin resistance were significantly increased in obesity control group, while the expressions of adiponectin mRNA and protein were obviously increased. Comparison with obesity control group, the expressions of adiponectin mRNA and protein, the concentration of plasma adiponectin were obviously increased, the concentration of blood glucose and insulin resistance were significantly decreased in obesity exercise group. Conclusion: Adiponectin mRNA and protein expression in visceral adipose tissue is decreased and accompanied by high blood glucose and insulin resistance in elderly obesity rats. Exercise can increase the adiponectin mRNA and protein expression in visceral adipose tissue, elevate levels of plasma adiponectin, and decrease the level of blood glucose and insulin resistance in elderly obesity rats.
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    Effect of lower-limb dominance and non-dominance shuttle runs under load carriage on the balance responses in young cadets
    MA Ji-zheng, WANG Zeng-gang, HU Fei, ZHAO Yan, YE Qiang, HUANG Qiang-nian, XU Sheng-jia
    CJAP. 2018, 34 (4): 350-354.   DOI: 10.12047/j.cjap.5634.2018.080
    Abstract   PDF (938KB) ( 99 )
    Objective: This test was designed to evaluate the effect of lower-limb dominance and non-dominance shuttle runs under load carriage during different exercise load at the same exercise intensity on the balance responses. Methods: Ten healthy young males were joined in this experiment, they were (20.80±2.04) years old and (173.99±2.87) cm tall. In a randomized cross-over design, they performed four times shuttle runs under unilateral load carriage:20 m×5 at dominant side, 20 m×5 at non-dominant side, 20 m×10 at dominant side, 20 m×10 at non-dominant side respectively. Balance abilities were evaluated immediately and 20 minute post-exercise respectively, and R-R interval was recorded. Results: The HR, EPOC and TRIMP for all exercise load were increased significantly after shuttle runs compared to rest (P<0.01), the EPOC and TRIMP for 20 m×10 shuttle runs were significantly higher than those for 20 m×5 shuttle runs at the dominant and non-dominant side (P<0.01). Compared to the rest, the instability indices (general, anteroposterior and mediolateral movement) for 20 m×5 and 20 m×10 shuttle runs immediately post-exercise at the dominant and non-dominant side were increased significantly (P<0.05), and the magnitude of the mediolateral movement for 20 m×10 shuttle runs was higher than that of 20 m×5 shuttle runs (P<0.05). While there were no obvious difference between the dominant and the non-dominant side (P>0.05), which showed symmetrical change. In addition, during 20 minute recovery, the balance ability for all exercise load was returned to the rest value (P>0.05). Conclusion: The shuttle runs could impair the trunk control ability immediately post-exercise, the magnitude of mediolateral movement was increased as the exercise load increased. The changes of balance responses were similar between the dominant and the non-dominant side, the dominant and the non-dominant side might show cross-effects.
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    Effect of oxidative stress on periprosthetic osteolysis induced by TCP wear particles in mouse calvaria and its mechanism
    LIU Jian-rong, MAO Hong-jiao, GUO Gao-li, FU Jing-lei, TONG Xia, QIAN Qin-qing, LUO Hua-gang, ZHANG Yun
    CJAP. 2018, 34 (4): 355-359.   DOI: 10.12047/j.cjap.5639.2018.081
    Abstract   PDF (1444KB) ( 69 )
    Objective: To explore the effect of oxidative stress on periprosthetic osteolysis induced by TCP wear particles in mouse calvaria and its mechanism. Methods: Thirty-six male ICR mice were randomly divided into three groups (n=12):sham group, TCP wear particles (TCP) group and N-acetyl-L-cysteine (NAC) group. Aperiprosthetic osteolysis model in mouse was established by implanting 30 mg of TCP wear particles onto the surface of bilateral parietal bones following removal of the periosteum. On the 2nd day post-operation, NAC (1.0 mg/kg) was locally injected to the calvarium under the periosteum every other day for 2 weeks. Then, all the mice were sacrificed to obtain blood and the calvaria. Periprosthetic osteolysis in the mouse calvaria was observed by tartrate resistant acid phosphatase (TRAP) staining; serum levels of tumor necrosis factor-alpha (TNF-α), interleukin-1beta (IL-1β), interleukin-6 (IL-6); total anti-oxidation capacity (T-AOC) and superoxide dismutase (SOD) activity were examined by ELISA and chemical colorimetry, respectively; protein levels of glucose-regulated protein 78 (GRP78), protein kinase R-like ER kinase (PERK), phospho-PERK (p-PERK), eukaryotic initiation factor 2α (eIF2α) and phospho-eIF2α (p-eIF2α) in periprosthetic bone tissue were detected by Western blot. Results: Compared with sham group, serum levels of TNF-α, IL-1β and IL-6, and osteolysis area were increased obviously in TCP group (P<0.05), and serum level of T-AOC and SOD activity were decreased significantly in TCP group (P<0.05), GRP78 expression, the ratio of p-PERK and PERK, p-eIF2α and eIF2α in the mouse calvaria of TCP group were up-regulated markedly. Compared with TCP group, serum levels of TNF-α, IL-1β and IL-6, and osteolysis area were decreased markedly in NAC group (P<0.05), serum level of T-AOC and SOD activity were increased obviously in NAC group (P<0.05), and GRP78 expression, the ratio of p-PERK/PERK and p-eIF2α/eIF2α were obviously down-regulated. Conclusion: Inhibition of oxidative stress can prevent periprosthetic osteolysis induced by TCP wear particles, which may be mediated by inactivation of PERK/eIF2α signaling pathway.
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    DNA methylation of HDAC4 gene affects the trans-differentiation process of hMSCs into sweat gland like cells
    ZHAO Huan-jun, DING Lu, FU Xiao-xiao, WANG Hui-ying, ZHANG Cui-ping
    CJAP. 2018, 34 (4): 360-363.   DOI: 10.12047/j.cjap.5584.2018.082
    Abstract   PDF (1319KB) ( 123 )
    Objective: To investigate the changesof DNA methylation in histone deacetylases 4 gene (HDAC4) and its effectduring the trans-differentiation process of human mesenchymal stem cells (hMSCs) into sweat gland like cells (SGLCs). Methods: Selected cell lines of human mesenchymal stem cells (hMSCs) were cultured and expended in vitro, the third generation ofhMSCs and heat-shocked sweat gland cells were picked up, and were co-culturedwith adding inducible factor in the transwell chamber. The sweat gland like cells (SGLCs)in experiment group and the hMSCs in control group were collected, the changes of DNA methylation degree of CpG dinucleotide sitesin histone deacetylases 4 gene (HDAC4) promotor were detected by methylation specific PCR (MSP)andMaldi-TOF Mass Array. And then, the hMSCs in experiment group were treated with 5-aza-CdR (5-aza-2-deoxycytidine, 10 μmol/L), while the hMSCsin control group were culturedwith PBS at the same time. ThemRNA expressions of HDAC4 gene and carcino-embryonic antigen (CEA)gene in the two groups were measured by RT-PCR. Results: The methylation of HDAC4gene in hMSCs was in high level before induction, the methylation degreeof CpG dinucleotide sites located in cg2463009 was 0.901, and the methylation degree of HDAC4gene in SGLCs was markedly decreased by 37% after induction, which was 0.531. The methylationlevel of CpG dinucleotide sites located in cg14823429was changed from 0.687to 0.386 after induction. The mRNA expression of HDAC4 gene was upregulated in test group after treated with 5-aza-CdR for 48 hours, the mRNA expression of CEA gene related with transdifferentiation was enhanced too at the same term, there was significantly statistic difference compared with control group (P<0.05). Conclusion: Methylation of HDAC4 gene participates in the regulation of the trans-differentiation of hMSCs into sweet gland like cells.
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    Effect of Angelica sinensis radix on colonic morphology and mucus secretion of experimental XuexuBianmi model mice
    DU Li-dong, REN Yuan, WU Guo-tai, NIU Ting-hui, WANG Zhi-wang, SHAO Jing
    CJAP. 2018, 34 (4): 367-370.   DOI: 10.12047/j.cjap.5616.2018.084
    Abstract   PDF (1243KB) ( 95 )
    Objective: To study the effects of Angelicaesinensis radix (Danggui) decoction on the therapeutic action and the colonic morphology and mucus secretion in XuexuBianmi model mice. Methods: Sixty Kunming mice, maleandfemaleinhalf, were randomly divided into six groups according to gender and weight (n=10):normal control group, XuexuBianmi model group, positive control group, and high-dose, middle-dose and low-dose Danggui groups. Except the normal control group, the mice in the rest groups were orally administrated with diphenoxylate (DPN) and subcutaneously injected with acetylphenyhydrazine (APH) and intraperitoneally injected with cyclophosphamide (CPA) to copy XuexuBianmi model. Fromthe 14th day, the mice in Danggui groups were orally administered with different doses of Danggui decoction (16.67、8.33、4.17 g/kg), the mice in positive control group were orally administered with Changtongshu granule (5 g/kg), the mice in XuexuBianmi model group were administered with normal saline (NS) at the same volume (10 ml/kg), once a day for consecutive 28 days. The general status were observed, the first black defecation time (FBDT), the water content in stool and in colon in mice of all groups were tested. And the colon tissue was stained with hematoxylin-eosin (HE) and AB-PAS to observe the changes of colonic morphology and the mucus secretion. Results: Compared with the normal control group, there appeared the XuexuBianmi syndromes in model group as follows, the defecation time (FBDT) was significantly prolonged, the water contentinstool and in colon were decreased (P<0.01), the colonic mucosa and gland were atrophied, mucous membrane layer was thinned (P<0.01), mucus secretion was decreased. Compared with the XuexuBianmimodel group, the Xuexu and Bianmi syndromes were improved, the FBDT was significantly shortened (P<0.05, P<0.01), the water content of colon was increased in the three groups of Danggui decoction (P<0.05, P<0.01). The water content of the stool was obviouslyincreased in groups of 16.67and 8.33kg.dDanggui decoction (P<0.05). The atrophy of colon mucosa and gland was improved, the mucus secretion was increased, and the colon lubrication function was improved in the three groups in different doses of Danggui decoction. Conclusion: Dangguican improve the mucosal atrophy, and increase the secretion of colonic mucus, then the stool is softened and promoted to discharge.
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    Antithrombotic effects of recombinant hirudin in mice and its mechanism
    CAO Wen-li, LI JING
    CJAP. 2018, 34 (4): 371-374.   DOI: 10.12047/j.cjap.5664.2018.085
    Abstract   PDF (927KB) ( 114 )
    Objective: To investigate the antithrombotic effects of recombinant hirudin and its mechanism. Methods: Sixty male Kunming mice were randomly divided into 6 group (n=10):control group, model group, aspirin (25 mg/kg) group, recombinant hirudinlow, middle and high dose (0.05, 0.1, 0.2 mg/kg) groups.Except mice in control group, 2.5 mg/kg carrageenan was injected intraperitoneallyto mice in the other groups to produce thrombosis on the mice tail. The mice in aspirin group were administrated intraperitoneally 25 mg/kg aspirin, the mice in recombinant hirudinlow, middle and high dose groups were administrated intraperitoneally 0.05, 0.1, 0.2 mg/kg combinanthirudin, the mice in control group and model group were administrated intraperitoneallynormal saline at the same volume respectively at 24 h, 0.5 h before injecting carrageenan and 24 h after injecting carrageenan. The black tail length of mice and the incidence of black tail were observed at 48h after injection of carrageenan; prothrombin time (PT), activated partial thromboplastin time (APTT), tissue plasminogen activator (t-PA), type-1 plasminogen activator inhibitor (PAI-1), 6-keto-PGF1α, and thromboxane B2 (TXB2) level in mice plasma were determined. Results: As compared with control group, the mice in model group presented tail thrombosis; PT level in plasma was significantly shortened (P<0.01), PAI-1 and TXB2levels in plasma were significantly increased (P<0.01), while the t-PA and 6-keto-PGF1α levels in plasma in model group were significantly decreased (P<0.01). As compared with model group, the thrombus length in the tail was significantly shortened (P<0.05, P<0.01), PT level was obviously prolonged (P<0.01), and the plasma levels of PAI-1 and TXB2 were significantly decreased (P<0.01), while the plasma levels of t-PA and 6-keto-PGF1α were significantly increased (P<0.01)in the mice of recombinant hirudin low dose, middle dose, high dose groups and aspirin group. As compared with aspirin group, the thrombus length in the tail was significantly increased (P<0.05), PT level was obviously shortened (P<0.01), and the plasma levels of PAI-1 and TXB2 were significantly increased (P<0.01)in the mice of recombinant hirudin low dose group; the plasma level of 6-keto-PGF1α was significantly decreased (P<0.01, P<0.05) in the mice of recombinant hirudin low dose and middle dose groups; the plasma levels of PAI-1 and TXB2 were significantly increased (P<0.01, P<0.05)in the mice of recombinant hirudin middle dose group. Conclusion: The recombinant hirudin can fight against thrombosis, its antithrombotic mechanisms may be related to its influence on the exogenous coagulation system and the promotion of fibrinolysis function.
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    Effect of matrine on tumor growth and inflammatory factors and immune function in Wistar rat with breast cancer
    ZHANG Rui-kui, WANG Chao
    CJAP. 2018, 34 (4): 375-378.   DOI: 10.12047/j.cjap.5657.2018.086
    Abstract   PDF (935KB) ( 147 )
    Objective: To study the effect of matrine on tumor growth, inflammatory factors and immune function in Wistar rat with breast cancer. Methods: Sixty female Wistar rats were randomly divided into control group (n=10) and the modeling group of breast cancer cell tumor-bearing rat (n=50), then the rats in modeling group were randomly divided into five groups (n=10):vehicle group, matrine low dose group (50 mg/kg), medium dose group (100 mg/kg), high dose group (200 mg/kg), and lentinan group (200 mg/kg). Except the control group, each rat in the other groups was subcutaneously inoculated 0.4 ml Walker 256 breast cancer cell suspension (5×107 cells/ml) in the right axillary. Each group was treated with corresponding drug by ig administration (10 ml/kg body weight) twice a day for 14 days. After 14 days, the blood sample was collected from ventral aorta, the tumor was removed and weighed to calculate tumor inhibitory rate. The levels of interleukin-2 (IL-2), interferon-γ (IFN-γ), interleukin-6 (IL-6), interleukin-10 (IL-10), transforming growth factor-β (TGF-β), CD3+, CD4+, CD8+, IgG, IgM, IgA in peripheral blood were determined. Results: The mean tumor weight of matrine low-dose, medium-dose, high-dose groups and lentinan group were (4.99±0.93) g, (4.52±0.92) g, (4.22±1.18) g and (4.52±0.92) g respectively, which were significantly lower than that in model group. There was no statistical difference on the mean tumor weight among matrine groups and lentinan group (P>0.05). After the drug intervention, the tumor inhibitory rates of matrine low-dose, medium dose, high-dose groups and lentinan group were 24.6%, 31.7%, 36.3%, and 27.9% respectively, there was no statistical difference among the four groups. The levels of IL-2, IFN-γ, CD8+ in vehicle group were lower than those of control group obviously (P<0.01), however, the levels of IL-6, IL-10, TGF-β, CD3+, CD4+, IgG, IgM, IgA were higher significantly than those of control group (P<0.01). The levels of IL-2, IFN-γ, CD8+ in matrine low-dose, medium dose, high-dose groups and lentinan group were higher than those of vehicle group obviously (P<0.01, P<0.05); while the levels of IL-6, IL-10, TGF-β, CD3+, CD4+, IgG, IgM, IgA were lower than those of model group markedly (P<0.01, P<0.05). The levels of IgM and IgA in matrine low-dose and medium-dose groups were higher than those of lentinan group obviously (P<0.01, P<0.05); the levels of IL-2, IFN-γ and IgA in matrine high-dose group were higher than those of lentinan group obviously (P<0.01, P<0.05); while the levels of IFN-γ in matrine low-dose group were lower than those of lentinan group markedly (P<0.05); the levels of IL-10 and CD4+ in matrine high-dose group were lower than those of lentinan group markedly (P<0.01, P<0.05). Conclusion: Matrine has an obvious antitumor action which is related to its ability to enhance cellular and humoral immunity, reduce inflammatory reaction.
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    Effects of combined administration of Radix Angelicae Sinensis and hydrocortisone on the therapeutic action in the murine asthma model
    WANG Zhi-wang, FU Xiao-yan, YAO Nan, GONG Zi-han, CHENG Xiao-li, DUAN Hai-jing, REN Yuan
    CJAP. 2018, 34 (4): 379-384.   DOI: 10.12047/j.cjap.5648.2018.087
    Abstract   PDF (1687KB) ( 137 )
    Objective: To investigate the effects of Radix Angelicae Sinensis (RASI) and hydrocortisone combination on the murine asthma model and the mechanism. Methods: BALB/c mice were randomly divided into control group, blood stasis model group, asthma model group, HSS group, RASI group and RASI+HSS group (n=12). Ovalbumin (OVA) was used to replicate mice asthma model and hydrocortisone sodium succinate (HSS) to copy blood stasis model. Effects of RASI, HSS and their combination on hemorheology, anti-asthma (asthmatic behaviors, lung function, lung index and water content in lung tissue) were observed. and anti-asthma mechanisms The expression of relative cytokines, high-mobility group box 1 (HMGB1), toll-like receptor 4 (TLR4) and nuclear factor-κB (NF-κB) was detected by ELISA and immunohistochemistry respectively. Results: Eight g/kg RASI, 0.05 g/kg HSS and their combination could significantly relieve asthma behavioral indicators, improve lung function, reduce lung index and water content in lung tissue, decrease the levels of TNF-α, IL-1β and IL-6 in broncho-alveolar lavage fluid (BALF), and inhibit the high expression of HMGB1, TLR4 and NF-κB in lung tissue. The improvement of lung function and the decrease in level of relative cytokines (TNF-α、IL-1β及IL-6) were better in RASI+HSS group than those in RASI group and HSS group, and the inhibition of protein expression of TLR4 and NF-κB was also too. Combined administration of RASI and hydrocortisone could decrease serum thromboxane B2 (TXB2) content and blood viscosity, which were increased induced by hydrocortisone. Conclusion: Combined administration of RASI and hydrocortisone have obvious anti-asthma effects and one of the mechanisms is to inhibit protein synthetization of HMGB1, TLR4 and NF-κB.The combined administration of RASI and hydrocortisone has stronger improvement of lung function than that of RASI and hydrocortisone alone, and it may be related to the inhibition of TLR4 and NF-κB synthetization. The combined administration of RASI can alleviate abnormal changes of hemorheology induced by hydrocortisone in treatment of asthma.
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  2019-03-15

Sponsor
Chinese Association for Physiological Sciences Academy of Military Medical Sciences Institute of Health and Environmental Medicine
Associate Sponsor
Institute of Basic Medical Sciences
Editor in Chief
WANG Hai
Edited and Published by
Editorial Board,Chinese Journal of Applide Physiology;Dali Dao,Tinanjin 300050,China



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