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Reproduction,genotype identification and evaluation of APP/PS1 transgenic mice
TAN Long, LI Hai-qiang, LI Yi-bo, LIU Wei, PANG Wei, JIANG Yu-gang
CJAP    2018, 34 (2): 111-114.   DOI: 10.12047/j.cjap.5541.2018.027
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Objective:To identify the genotype of (APP/PS1) transgenic mice and evaluate the changing of cognitive and behavioral fu nctions, provide an effective animal model for the Alzheimer's disease (AD) research. Methods:Male APP/PS1 transgenic mice mated with female APP/PS1 transgenic mice, and the genotype of their filial mice was identified by PCR. The APP +/PS1 + mice were assigned into AD model group (AD group, n=8), and the APP/PS1 mice were assigned into control group (CT group, n=8). The Morris water maze test was carried out to detect the capacity of learning and memory of mice. After that, the mice were sacrificed and the brain tissues were sampled and stained by HE and congo red for the pathological examination. Results:①A APP/PS1 genome DNA about 360 bp size was detected. The methods of feeding and breeding were successful to attain APP/PS1 transgenic mice.②Statistical significance was found in the differences of the capacity of learning and memory between 7-month-old APP/PS1 positive mice and negative mice (P<0.05).③The results of HE stain showed that the structure and cellular morphology of hippocampus of AD mice were obviously abnormal. The results of congo red stain showed that positive amyloid plaque was observed in brains of AD mice. Conclusion:APP/PS1 transgenic mice present typical symptoms and behaviors of Alzheimer's disease. The transgenic mouse is an effective tool for the research and prevention of AD.
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Selection of modeling time for type 2 diabetes mellitus mouse
YANG Zhang-liang, TONG Hai-ling, SUN Meng-die, YUAN Jie, HU Ying, WANG Xu-tao, QI Min-you
CJAP    2019, 35 (2): 155-159.   DOI: 10.12047/j.cjap.5728.2019.034
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Objective: To analyze the changes of blood biochemical index and the pathological changes of myocardium and kidney in type 2 diabetic mouse at different time points, which can provide the basis for the selection of type 2 diabetic modeling time for later research. Methods: After 6 weeks of feeding with high-fat diet, 24 healthy male ICR mice were injected with streptozocin (STZ, 30 mg/kg) intraperitoneally for 5 days to establish diabetic models. After 9 days, a random blood glucose ≥ 11.1 mmol / L was measured as diabetic mice. 4, 6 and 8 weeks after successfully preparing the diabetic mouse, 8 diabetic mice (a group)would be sacrificed each time. Then the biochemical and pathological conditions were analyzed: ① the indexes of heart and kidney were calculated. ②the serum levels of creatine kinase (CK), lactate dehydrogenase (LDH), creatinine (Cr) and blood urine nitrogen (BUN) were determined. ③ Histopathological changes of myocardium and renal tissues were observed by hematoxylin and eosin (HE) staining. Masson staining was used to observe the fibrosis of myocardium. PAS staining was adopted to observe the pathological changes of renal tissue. In addition, 8 ICR male mice were taken as the control group. Results: At the 4th, 6th and 8th week, cardiac organ coefficient, the values of LDH and CK were all increased compared with the control group. Cardiomyocyte hypertrophy and myocardial fibrosis could be observed. Renal organ coefficient, the values of Cr and BUN were increased. Glomerular hypertrophy, basement membrane thickening and atrophy could be perceived. Conclusion: At the 6th week, related biochemical and pathological changes in diabetic mice were comparatively obvious and breeding time was relatively short. Thus, 6 weeks after the preparation of the diabetic mice would be the optimal time for type 2 diabetes mellitus modeling, proper for inventions of drugs and other research purposes including pathology, physiology, biochemistry, etc.
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The research of hemoglobin mass on athletic biological passport for blood doping detection
WANG Jun, ZHANG Mei-ling, WANG Song-li, LIU Min, YANNIS Pitisladis
CJAP    2018, 34 (6): 519-523.   DOI: 10.12047/j.cjap.5701.2018.116
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Objective: To research the changes of blood parameters on different time points after micro-rHuEPO injection on young men and hope to provide evidences for Athletic Biological Passport (ABP) using in the detection of blood doping.Methods: Fourteen health young men were injected with micro-rHuEPO for 7 weeks, twice per one week.The subjects were treated with ferralia 105 mg every day during the injections.The control group was administrated with the same volume of saline solution injection and placebo 105 mg.The blood parameters (red blood cell, hemoglobin, reticulocyte, total hemoglobin, plasma volume, etc) were tested 11 times (7 days before the first injection, 3, 10, 17, 24, 31, 38 and 45 days after the first injection, 1, 2 and 3 weeks post the seventh week of last injection).By analyzed these total and concentration parameters, hope to know the effective of two kinds of parameters on ABP.Results: The levels of RBC and[Hb] were increased sharply after two weeks of injection and reached the peak in 5-6 weeks (9-10%, P<0.01).It sustained till 3 weeks after the last injection.Total hemoglobin was increased significantly during the whole procedure of injections and reached the peak in week 5 (10%, P<0.01) till 1 week after the last injection.Red blood cell volume was increased with the EPO injection and reached the peak in week 5 (P<0.01), but blood volume wasn't increased significantly.Plasma volume was decreased with EPO injection and it was hemoconcentration.Conclusion: It can enhance the total and concentration parameters through 7-week micro-rHuEPO injections and the total haemoglobin is more sensitive.So tHb can be used to detect blood doping in ABP.At the end of the last injection, the total blood parameters, such as the total hemoglobin, returned to normal, while the concentration index remained highly, possibly as a result of blood concentration.
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Prognosis-related miRNA bioinformatics screening of lung adenocarcinoma and its clinical significance
ZHANG Hao-min, YANG Bo, CHEN Hong-fei, CHI Xiao-hua, XI Yi-bo, CHEN Xi-meng, GUO Bin, HE Pei-feng, LU Xue-chun
CJAP    2018, 34 (6): 530-535.   DOI: 10.12047/j.cjap.5755.2018.118
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Objective: To investigate the prognosis-related miRNA histological features and clinical significance of lung adenocarcinoma. Methods: Using The Cancer Genome Atlas (TCGA) data, the miRNA expression profile data of human lung adenocarcinoma were searched for differential analysis, and the prognosis-related miRNAs were screened by Cox risk regression model. The targeted miRNAs were predicted by mirwalk analysis platform, KEGG functional enrichment analysis, and finally, predict the function of prognosis-related miRNAs. Results: A total of 46 differential miRNAs in lung adenocarcinoma were screened, including 19 up-regulated and 27 down-regulated. Six prognostic-related miRNAs were screened by Cox survival analysis, namely hsa-mir-21, hsa-mir-142, hsa-mir-200a high expression, hsa-mir-101, hsa-let-7c, hsa-mir-378e low expression, hsa-mir-21 and hsa-mir-378e were associated with poor prognosis in patients with lung adenocarcinoma, and the survival time was shortened significantly (P<0.05, AUC=0.618). KEGG analysis showed that the above prognosis-related miRNA targeting regulatory genes were related with immune response pathways, miRNA and cancer pathways, metabolic pathways and so on. Conclusion: Hsa-mir-21 and hsa-mir-378e are associated with poor prognosis of lung adenocarcinoma, and may be used as a molecular marker for prognosis of lung adenocarcinoma after further clinical verification.
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CJAP    2019, 35 (2): 113-115.   DOI: 10.12047/j.cjap.5776.2019.025
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How to follow the four principles of experimental design in physiological research
HU Wan, SUN Yan-song, HU Liang-ping
CJAP    2017, 33 (1): 89-93.   DOI: 10.12047/j.cjap.5448.2017.023
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Objective: To offer a series of efficient methods to physiological researchers in following the four principles of experimental de-sign (hereinafter referred to as four principles).Methods: We expounded the concrete way of following the four principles through giving an outline of the four principles, elaborating the common mistakes and cases in the applications of four principles in physiological research and in-troducing briefly how to follow the four principles in physiological research.Results: We should choose the appropriate randomization method according to the actual situation, usually stratified randomization was a good way. We should strive to set reasonable control group to reflect the value of control principle veritably. Giving the appropriate sample size for each group rightly was the basic guarantee for the research result could be reproduced. Following the principles of randomized, controlled and repetition strictly could achieve an excellent balance between con-trol groups ultimately, that was to say that the balanced principle was a principle of gatekeeper.Conclusion: Following the four principles strictly is critically important to assure the result accurately and reliably in physiological research.
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Analysis of high-dose dexamethasone in different cycles on efficacy and safety of newly diagnosed ITP
XIAO Yue, LI Tang-fei
CJAP    2018, 34 (5): 436-440.   DOI: 10.12047/j.cjap.5673.2018.099
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Objective: To compare the efficacy and safety of high-dose dexamethasone in treating new-diagnosed primary immune thrombocytopenia (ITP) in monocycle, di-cycle, tri-cycle. Methods: Divided by the ratio of 1:1:1, 93 newly diagnosed patients were randomly accepted monocycle (Group A:dexamethasone 40 mg once a day, from day1 to day4), dicycle (Group B:dexamethasone 40 mg once a day, from day 1 to day 4, day 15 to day 18), tri-cycle (Group C:dexamethasone 40 mg once a day, from day 1 to day 4, day 15 to day 18, day 29 to day 32) of high-dose dexamethasone treatment. Its efficacy and safety on the patients in three groups were compared. Results: Ninety-three newly patients with new-diagnosed ITP were divided into Group A, B, and C, 31 patients in each group. In terms of short-term benefits, there was no statistically significant difference among the 7th and 14th day complete response rate after end of treatment. However, there was statistically significant difference after the end of treatment on the 7th day response rate (41.9% vs 70.0% vs 90.0%, P<0.01) and the 14th day response rate (16.1% vs 36.70% vs 63.3%, P< 0.01); in terms of long-term benefits, there was no statistically significant difference among the 120-day response rate, the complete response rate within the treatment on the 60th, 90th and 120th day and the relapse rate at 90th and 120th day; however, there was statistically significant difference among the 60- day response rate (10.0% vs 26.6% vs 53.3%, P<0.01), 90-day(0.0% vs 13.3% vs 30.0%, P<0.01) and 60-day relapse rate(88.9% vs 73.3% vs 46.7%, P<0.01). Mostly of the treatment-related adverse reactions in the three groups were mild, and most patients are tolerable. Conclusion: Although the complete response rate of ITP patients did not improved by increasing the cycle of high-dose dexamethasone, but improved response rate in three months, and adverse reactions were tolerable, which could be used as a reference for clinical use.
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Effects of deoxygedunin on Alzheimer-like pathologic dysfunction induced by D-galactose combined with AlCl3
CHEN Jian-guo, JIANG Qi-chuan, WEN Bo, WANG Ruo-ya, WU Ya-geng, LI Xiang
CJAP    2018, 34 (6): 496-500.   DOI: 10.12047/j.cjap.5732.2018.111
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Objective: To investigate the effects of Deoxygedunin on Aβ deposition, learning memory, and oxidative stress induced by D-galactose combined with AlCl3 in model rats with Alzheimer's disease and its possible mechanism.Methods: Male SD rats were randomly divided into three groups (n=12):control group, model group (AD) and intervention group (AD+Deo). Morris water maze test was used to detect learning/memory and cognitive function in rats.Glutathione peroxidase (GSH-Px), superoxide dismutase (SOD) and malondialdehyde (MDA) contents in homogenate of hippocampus were detected by enzyme-linked immunosorbent assay (ELISA).Tau protein expression in rat cerebral cortex was detected by immunohistochemistry.Western blot was used to detect the expressions of extracellular signal regulated kinase 1(ERK1), protein kinase B (PKB) and tropomyosin-related kinase B (TrkB) on TrkB signaling pathway.Results: The results of water maze test showed that D-galactose combined with AlCl3 induced a significant increase in the escape latency compared with the control group (P<0.05).Deoxygedunin could reverse the increase of the escape latency of the model group (P<0.05).On the 7th day after removal of the platform, the model group showed an increase in escape latency compared with the control group and the intervention group (P<0.01), and the number of crossing platforms was declined (P<0.05); The results of immunohistochemistry and ELISA showed that the expressions of Aβ and tau protein in the model group were increased significantly compared with those of the control group (P<0.01).The activities of SOD and GSH-Px were decreased significantly and the content of MDA was increased significantly.Compared with the model group, Deoxygedunin could reverse the increase of the expressions of Aβ and tau protein (P<0.01), the decrease of SOD and GSH-Px activities (P<0.05) and the increase of the MDA content (P<0.05).Western blot results showed that Deoxygedunin treatment reversed the decreased phosphorylation levels of TrkB, AKT and ERK1 in hippocampus of the model group.Conclusion: Supplement of Deoxygedunin can significantly reverse Aβ deposition, oxidative stress and cognitive deficits by activating the TrkB signal transduction pathway, which suggest that Deoxygedunin may serve as a promising therapeutic candidate for attenuating AD-like pathological dysfunction induced by D-galactose combined with AlCl3.
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Effects of metformin on depressive behavior in chronic stress rats
LI Gai-fen, ZHAO Ming, ZHAO Tong, CHENG Xiang, FAN Ming, ZHU Ling-ling
CJAP    2019, 35 (3): 245-249.   DOI: 10.12047/j.cjap.5775.2019.052
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Objective: To detect the effects of metformin on the depressive-like behaviors in rats. Methods: Forty male SD rats were randomly divided into four groups: control group (CON group), metformin group (MET group), model group (CUMS group), model + metformin group (CUMS + MET group), 10 rats in each group. Chronic unpredictable mild stress (CUMS) method was used to establish rat depression model in three weeks. After the model was established successfully, two metformin groups were intraperitoneally injected with metformin (100 mg/kg), while the control group and the model group were injected with the same amount of saline once a day for two weeks. After that, the changes of weight gain, sucrose water preference experiment, forced swimming test, tail suspension immobility test and open field test were detected. The morphological changes of hippocampus were observed by Nissl staining. Results: Compared with the control group, the weight gain of rats in CUMS group was significantly slowed down (P<0.05), the sucrose preference rate and the spontaneous activity were significantly reduced (P<0.05), and the immobility time in forced swimming and tail suspension immobility test was significantly prolonged (P<0.05), and the morphological structure of hippocampus was changed, which confirmed the success of CUMS depression model. Compared with CUMS group, metformin treatment had no significant effect on body weight of rats, but it could significantly improve sucrose water intake, immobility time and spontaneous activity of CUMS depression model rats (P<0.05), and improve the abnormal morphological changes of hippocampus in CUMS rats. Conclusion: Metformin has a therapeutic benefit against CUMS-induced depression, which provides a new treatment for patients with diabetes mellitus complicated with depression.
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Protective effect of Ginkgo biloba extract on paracetamol-induced acute hepatic injury in mice
ZHANG Quan-shu, WANG Xiang-peng, XIE Yan-ni, WU Lu-lu, LIU Hong
CJAP    2018, 34 (5): 432-435.   DOI: 10.12047/j.cjap.5655.2018.098
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Objective: To investigate the protective effects of Ginkgo biloba extract(GBE) on paracetamol(APAP)-induced acute hepatic injury in mice and its mechanism. Methods: Thirty mice were randomly divided into control group, model group, GBE low, medium and high-dose(50,100,and 200 mg·kg-1)groups,with 6 mice in each group. All mice except control group were administered with APAP(300 mg/kg)for one time by intraperitoneal injection. The mice in GBE low, medium and high-dose groups were intragastric administered with GBE for 2 d consecutively, then samples were harvested for analysis. The appearance and pathology of liver were observed. The levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in serum and the levels of superoxide dismutase (SOD), myeloperoxidase(MPO), glutathione (GSH) and malondialdehyde (MDA) in hepatic tissue were measured. Western blot was used to detect the protein expressions of Nrf2 and HO-1. Results: Compared with control group, in model group, the appearance and pathology of liver were bad, the levels of ALT,AST,TNF-α and IL-6 in serum were increased significantly(P<0.01),the levels of GSH and SOD were decreased while the levels of MDA and MPO were increased in hepatic tissue(P<0.01), the expressions of Nrf2 and HO-1 were increased in hepatic tissue(P<0.05). Compared with model group, in GBE groups, the appearance and pathology of liver were improved, the levels of ALT,AST,TNF-α and IL-6 in serum were decreased significantly(P<0.01), the levels of GSH and SOD were increased while the levels of MDA and MPO were decreased in hepatic tissue(P<0.01), the expression of Nrf2 and HO-1 were increased in hepatic tissue(P<0.05). The high-dose of GBE possessed the most obvious treatment effect among them. Conclusion: GBE may play a protective role in APAP-induced acute hepatic injury through Nrf2/HO-1 pathway.
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CJAP    2018, 34 (5): 461-463.   DOI: 10.12047/j.cjap.5718.2018.104
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The effects of 10-week core strength training on the body balance in the middle-aged men
LIU Shan-yun, ZHANG Xiang, SUN Jiang-bo, LI Qian-nan, LIU Yue, LIAN Zhi-qiang, ZHANG De-rong
CJAP    2017, 33 (4): 365-368.   DOI: 10.12047/j.cjap.5482.2017.088
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Objective: To observe the effects of core strength training on the body balance of middle-aged men, which could provide evidence for improving the body balance and reducing the risk of falls in the middle-aged men.Methods: Sixteen 50~60 years old men were randomly divided into experimental group (core strength training, n=8) and control group(n=8). Intermittent core strength trainings were used for dynamic and static training, push-pull training and unarmed lower limb strength training by suspension ropes and yoga mat. The time for training was 50~60 min/day, 4~5 times/week for a total of 10 weeks. The men in control group remained their original living habits. We measured the ability of body static balance and dynamic balance before and after the core strength training.Results: ① Compared with before exercise training, both closed single foot standing time and the body dynamic balance were increased significantly in the experimental group after exercise training(P<0.01)(10.63±1.69 s vs 9.00±2.27 s; 77.38±10.94 vs 89.50±5.53). ② Compared with before exercise training, star excursion balance test(SEBT) values were significantly increased in the left leg(the right leg support) in eight directions and the right leg(the left leg support)in six directions (P<0.01).Conclusion: Ten-week core strength training can significantly increase the body static and dynamic balance in the middle-aged men.
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Effects of Dandelion polysaccharide on IL-6/STAT3 signaling pathway in ulcerative colitis rats
WANG Qian, BIE Yu-long, WANG Dou, FAN Wen-tao
CJAP    2017, 33 (5): 422-425.   DOI: 10.12047/j.cjap.5525.2017.102
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Objective: To investigate the effects of dandelion polysaccharide on IL-6/STAT3 signaling pathway in rats with ulcerative colitis.Methods: Forty SD rats were randomly divided into four groups (n=10):control group, model group, positive control group and dandelion polysaccharide group. The ulcerative colitis model was established by treated with 2,4,6-trinitrobenzene sulfonic acid (TNBS). The positive control group was treated with mesalazine 10 mg/kg·d and dandelion polysaccharide group was treated with dandelion polysaccharide 10 mg/kg·d. The levels of interleukin -6(IL-6), colonic myeloperoxidase (MPO) and interleukin-6 receptor (sIL-6Rα) were measured after 4 weeks of treatment. The pathological changes of colonic mucosa were observed in rats. The gene expressions of glycoprotein 130 (gp130), transcriptional activator3(STAT3) and IL-6 were detected.Results: Compared with the normal control group, the level of serum IL-6 in the model group was significantly higher than that in the control group (P<0.01). Compared with the model group, the serum levels of IL-6 in the dandelion polysaccharide group and the methacetin group were significantly decreased (P<0.01). Compared with the model group, the MPO positive density of the rats in the dandelion polysaccharide group and the methacetin group was significantly lower than that in the normal group (P<0.01). Compared with the model group, the levels of sIL-6Rα and gp130 in the rats were significantly lower than those in the model group (P<0.01). Compared with the model group, the expressions of STAT3 and IL-6 mRNA in the intestinal tissue of the rats in the dandelion polysaccharide group and the methacetin group were decreased significantly.Conclusion: Dandelion polysaccharide can decrease the level of IL-6 in rats with ulcerative colitis, regulate the expression of sIL-6Rα and gp130 protein in IL-6/STAT3 pathway, and then down-regulate the expressions of STAT3 and IL-6 mRNA in intestinal tissue of rats, alleviate the colon inflammation state, protect and repair the mucosal tissue. Dandelion polysaccharide plays a role in the treatment of ulcerative colitis.
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Effects of Health Qigong Baduanjin on diabetes related indexes in middle-aged obese women
LIU Tao, BAI Shi, ZHANG Rong-chao
CJAP    2018, 34 (1): 19-22.   DOI: 10.12047/j.cjap.5484.2018.006
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Objective: To investigate the effect of Health Qigong Baduanjin on the related indexes of obese middle aged women with diabetes and to provide new ideas for the intervention treatment of diabetes.Methods: A total of 40 middle-aged female obese diabetic patients were randomly divided into the control group and the exercise group(n=20), the age was(57.2±5.4) years old. Fitness training group performed eight new Baduanjin exercises for 24 weeks of intervention, the control group did not exercise, body weight, waist circumference, body mass index (BMI), waist hip ratio (WHR), fasting blood glucose (FPG), glycosylated hemoglobin (HbAlc), triglyceride(TG), total cholesterol (TC), low density lipoprotein (LDL), high density lipoprotein (HDL) levels and serum retinol binding protein 4(RBP4) index were observed in the two groups.Results: After exercise, the waist, WHR, FPG, TG, HbAlc, HDL and RBP4 levels of the the patients in the experimental group were decreased significantly compared with those of before exercise and those of the patients in the experimental control group before and after exercise (P<0.05).Conclusion: Health Qigong Baduanjin can reduce the blood sugar of obese female patients with diabetes, and has some improvement effect on the body part of obesity and blood lipid indicators.
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Effects of Ringer’s solution with different concentrations of alcohol on biphasic compound action potentials of frog sciatic nerve trunk
HUANG Zhi-hua, WEI Pei-jian, JIANG Ling, CHEN Sui, CHENG Bi-hong, LIN Ying, WU Lin-geng, XU Qiu-xiong, WU Shao-wei, WANG Hai-yan, SHEN Jian-xin
CJAP    2019, 35 (3): 232-238.   DOI: 10.12047/j.cjap.5750.2019.050
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Objective: To quantitatively investigate the effects of Ringer’s solution with different concentrations of alcohol (1%~80%) on biphasic compound action potentials (AP) from frog sciatic nerve trunk, and their recoveries from alcohol effects. Methods: Individual segments of frog sciatic nerve trunk with a length of 6 to 8 cm were prepared. Ringer’s solution with different concentrations of alcohol (0%, 1%, 2%, 4%, 8%, 16%, 32%, 48%, 64% and 80%) was applied onto the segment of the trunk between the stimulus and ground electrodes via an agent reservoir which was newly armed in a nerve trunk shielded chamber for 5 minutes. The nerve trunk was respectively electro-stimulated to generate the biphasic compound AP which was recorded using the experimental system of BL-420F. This was followed by 5 times washout plus 5 min administration with Ringer’s solution before recovery recording of AP. Results: Compared to normal Ringer’s solution, Ringer’s solution with alcohol at ≤4% did not have dramatic impacts on the AP amplitude and conduction velocity, while Ringer’s solution with alcohol at ≥8% there was significant decrease in these two parameters. Ringer’s solution with alcohol at the conentrations of 16%, 32% and ≥48% could prevent a small proportion (30%), a large proportion (90%) and all (100%) of sciatic nerve trunks, respectively, from generating AP. Washout with normal Ringer’s solution after alcohol application at the concentration of ≤32%, AP could totally recover to normal status. While alcohol at the concentration of 48%, 64% and 80%, the probabilities to regenerate APs were 90%, 40% and 0%, and the AP amplitudes were decreased to 60%, 36% and 0%, respectively. After washout, AP conduction velocity showed no difference with alcohol at the concentration of ≤8% when compared with that before washout, while it could not be recovered to normal under alcohol at ≥16%. Conclusion: Ringer’s solution with different concentrations of alcohol exerts different effects on biphasic compound AP amplitude and conduction velocity. Hopefully, our findings could be helpful for the alcoholic usage and its recovery from alcoholic damage.
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The protective effect of tea polyphenols on chronic alcoholic liver injury in rats
ZHANG Yong, LI Miao-miao, HUA Tian-miao, SUN Qing-yan
CJAP    2018, 34 (6): 481-484.   DOI: 10.12047/j.cjap.5708.2018.107
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Objective: To investigate the protective effects and potential mechanisms of tea polyphenols intervention on excess alcohol intake induced liver injury in rats. This study established the animal model of chronic liver injury rats induced by alcohol. Our results will provide experimental evidence for the effects of tea polyphenol on chronic alcoholic liver injury. Methods: Alcohol-induced liver injury rat models were established, and the tea polyphenols intervention was performed in the meantime. After 8 weeks, rats were anesthetized, and visceral fat and liver samples were separated, weighted and stored. Visceral fat content was evaluated in fat/body weight ratio. Liver lipid accumulation was assessed by liver index and the result of Oil Red O staining. Hepatic superoxide dismutase (SOD) activity, malondialdehyde (MDA) content, total antioxidant capacity assay (T-AOC) and glutathione peroxidase (GSH-Px) activity were detected. And fatty acid translocase (FAT/CD36) protein level in liver was detected. Results: Compared with the control group rats, the fat/body weight ratio, SOD/MDA, T-AOC and GSH-Px activity of chronic liver injury rats were decreased significantly (P<0.05,P<0.01). Meanwhile the liver index, FAT/CD36 protein level and lipid deposition in liver of chronic liver injury rats were increased (P<0.01). Compared with chronic liver injury rats, the tea polyphenols intervention increased fat/body weight ratio (P<0.05), and significantly increased SOD/MDA, T-AOC and GSH-Px activity (P<0.01). Meanwhile the tea polyphenols intervention reduced liver index (P<0.01), FAT/CD36 protein level (P<0.01) and lipid deposition in liver. Conclusion: Tea polyphenols intervention can improve lipid deposition and oxidative stress in chronic alcoholic liver, which is concurrent with decreased FAT/CD36 protein expression on the hepatocyte membrane.
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Changes of apoE protein expression in lung of mice with hypoxic pulmonary arterial hypertension
LIU Ling-yan, HUANG Tian-peng, HAO Jia-le, CHEN Ran, FAN Xiao-fang, GONG Yong-sheng, MAO Sun-zhong
CJAP    2018, 34 (6): 492-495.   DOI: 10.12047/j.cjap.5723.2018.110
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Objective: To observe the changes of apolipoprotein E (apoE) protein expression of pulmonary tissue in mice with pulmonary hypertension induced by hypoxia. Methods: The animal model of hypoxic pulmonary hypertension was established by exposing the mice to isobaric hypoxic chamber for 3 weeks (23 h/d, regular chow feed).Twenty male wild type (WT) C57BL/6 mice and twenty apoE gene knockout (apoE-KO) mice were randomly divided into normoxia group and hypoxia group. The plasma concentrations of low density lipoprotein (LDL), high density lipoprotein (HDL) and total cholesterol were detected by ELISA method. The protein expression of apoE in lung and liver, and peroxisome proliferators-activated receptor gamma (PPARγ) in lung were measured by Western blot.Results: ①In WT mice, the right ventricular systolic pressure (RVSP) and the weight ratio of right ventricle (RV) to left ventricle plus septum (LV+S) of hypoxia group were significantly higher than those of normoxia group by 68% and 59% (P<0.05), respectively. The plasma concentration of HDL and HDL/LDL of hypoxia group were significantly lower than those of normoxia group by 17% and 40% (P<0.05), respectively.The protein expression of apoE in lung and in liver of hypoxia group were significantly down-regulated than those of normoxia group by 48% and 52% (P<0.05), respectively.The protein expression of PPARγ in lung was significantly down-regulated than that of normoxia group by 37%(P<0.05).RVSP were significantly negative correlated with the protein levels of apoE and PPARγ in lung (P<0.01).② In apoE-KO mice, RVSP and the weight ratio of RV to LV+S of hypoxia group were significantly higher than those of normoxia group by 96% and 86% (P<0.05), respectively.RVSP and RV to (LV+S) of hypoxia group in apoE-KO mice were significantly higher than those of hypoxia group in WT mice by 29% and 24% (P<0.05), respectively.Conclusion: Down-regulated expression of apoE in lung tissue participates in the pathological proceeding of pulmonary hypertension induced by hypoxia.
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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
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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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Effect of Beclin-1 shRNA on hypoxia-induced autophagy in SH-SY5Y cells
LU Na, LI Cheng-zhang, LUO Xiao-qiu, YANG Kun-li
CJAP    2018, 34 (5): 464-469.   DOI: 10.12047/j.cjap.5661.2018.105
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Objective: To observe the effect of hypoxia on autophagy in Beclin-1-knockdown SH-SY5Y cells by constructing a stable transfected SH-SY5Y cell lines of silencing Beclin-1 gene. Methods: Beclin-1shRNA lentiviral vector and negative control lentiviral vector were constructed; the vector was transfected into SH-SY5Y cells; then the expression of Beclin-1 mRNA was detected by RT-PCR, the level of Beclin-1 protein was detected by Western blot. CCK-8 method was used to determine the effect of Beclin-1 knockdown on the viability of SH-SY5Y cells. Next, the blank control, negative control and transfected cells were cultured under 21% normoxia and 5% hypoxia conditions. The expression of LC3 protein in each group was detected by Western blot and the autophagic bodies were observed by electron microscopy. Results: Beclin-1 shRNA significantly inhibited the expression of Beclin-1 mRNA and protein in SH-SY5Y cells; after silencing Beclin 1 gene, the survival rate of Beclin-1 shRNA group cells was no different from that of negative control (NC) group. After 5% hypoxia treatment, compared with NC group, the ratio of LC3Ⅱ/LC3Ⅰand the number of autophagy bodies were all decreased in Beclin-1 shRNA group. Conclusion: Beclin-1 knockdown SH-SY5Y cell lines and negative control cell lines were successfully established. Lentivirus-mediated Beclin-1 shRNA has no effect on the viability of SH-SY5Y cells, but can inhibit hypoxia-induced autophagy.
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Effects of different steps of brisk walking on the fitness of middle-aged and old men
FAN Bin
CJAP    2018, 34 (2): 126-129.   DOI: 10.12047/j.cjap.5605.2018.000
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Objective:To analyze the effect of walking pace on the fitness of middle-aged and elderly people in order to provide more theoretical data for the development of entensive mass fitness based on the means of sports physiological and biochemical techniques. Methods:The selected 80 middle-aged and elderly subjects were divided into three groups:brisk walking group A (n=30, 56.26 ±3.68 years), brisk walking group B (n=30, 57.65 ±4.78 years), and control group C (n=20, 55. 73 ±4. 18 years). Exercise group A:10 000~12 000 steps/day, a total of 16 weeks; walking group B:10 000~12 000 steps/day in the first 10 weeks, 14 000~15 000 steps/day in the last 6 weeks. Control group C:keeping the normal living conditions. The indicators were tested before the experiment and after the tenth week and the sixteenth week of the experiment. Test indicators included systolic blood pressure, diastolic blood pressure, heart rate, vital capacity, timed vital capacity, total cholesterol, triglycerides, high density lipoprotein cholesterol, low density lipoprotein cholesterol. Results:The levels of systolic blood pressure, diastolic blood pressure, total cholesterol and triglyceride were decreased significantly (P<0.05, P<0.01), while the vital capacity and high density lipoprotein cholesterol were increased significantly in both exercise groups afler 10 and 16 weeks compared with those in the same group betor test(P<0. 05, P<0. 01). The levels of diastolic blood pressure, timed vital capacity, serum triglyceride and high density lipoprotein cholesterol in exercise group B after 16 weeks were significantly higher than those of exercise group A (P<0.05). Conclusion:Ten thousand steps away every day can effectively improve the indexes of blood pressure, vital capacity, serum triglyceride, total cholesterol and high density lipoprotein cholesterol of middle-aged and elderly men and it can further improve the indicator above by increasing the number of steps after 10 weeks.
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Anti-hepatoma effects of Smac analogue Birinapant and its related molecular mechanism
JIANG Pan-ruo, KE Rui-jun, ZHU Ming-liao, LOU En-zhe, XIE Jia-geng, CHEN Jia-yu
CJAP    2018, 34 (6): 524-529.   DOI: 10.12047/j.cjap.5685.2018.117
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Objective: To investigate the effects of Birinapant on hepatocellular carcinoma cells and its related molecular mechanisms.Methods: Human hepatocellular carcinoma cells QGY-7701 were treated with 0, 1, 5, 25 and 125 nmol/L Birinapant for 24, 48 and 72 hours respectively, each experiment 3 wells.The proliferation activity of cells, the apoptosis levels, the cells nuclear type, the mitochondrial membrane potential, the transcription and expression levels of genes and the cytotoxicity of Birinapant were analyzed.At the same time, 4-week-old male BALB/C mice were randomly divided into 5 groups, with 20 mice in each group.The mice were inguinal injected with QGY-7701 cells, and then subcutaneous injected with Birinapant (concentrations ranging from 0, 1, 5, 25, 125 μg/kg) in each group after two days, once every other day.On 18th day since first Birinapant injection, 10 mice were killed in each group to weigh tumor tissue and survival time was recorded from the remaining 10 mice.The effects of Birinapant on the growth of the tumor and the survival time of tumor-bearing mice were observed.Results: Compared with the negative control (NC) group, the proliferation activity of QGY-7701 was inhibited significantly after Birinapant treatment and the apoptosis levels were increased significantly (P<0.01).The cell mitochondrial membrane potential was decreased and the karyotype was changed (P<0.01).At the same time, the transcription and expression levels of genes cellular inhibitor of apoptosis protein 1(cIAP-1), cellular inhibitor of apoptosis protein 2(cIAP-2), ras, raf, mek and erk were significantly decreased (P<0.01), while the expression levels of caspase-3 and caspase-9 genes were up-regulated (P<0.01).Compared with the model group (MG), the growth of the tumor was inhibited significantly and the survival time of the tumor-bearing mice was prolonged after Birinapant treatment (P<0.01).Conclusion: Birinapant can inhibit the expression of cIAP-1, cIAP-2 and the proteins of Ras-Raf-MEK-ERK signal pathways, so as to activate the mitochondria mediated endogenous apoptosis pathway.Birinapant shows a certain inhibitory effect on liver cancer.
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Injury of rat blood vessels caused by acute ozone exposure and its mechanism
YANG Hu, LI Ning, HAN Jie, ZHU Chen-li, TIAN Lei, LIN Ben-cheng, XI Zhu-ge, LIU Xiao-hua, CHU Nan
CJAP    2019, 35 (3): 193-198.   DOI: 10.12047/j.cjap.5748.2019.042
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Objective: To investigate the vascular damage effects and possible mechanism of acute exposure to ozone (O3) in male Wistar rats. Methods: One hundred and twenty male Wistar rats were randomly divided into six groups, 20 in each group. The experimental animals were placed in a gas poisoning cabinet, the control group was exposed to filtered air, and the treatment group was exposed to ozone at concentrations of 0.12 ppm, 0.5 ppm, 1.0 ppm, 2.0 ppm, and 4.0 ppm, respectively, for 4 hours. Arterial blood pressure data were obtained by PC-lab medical physiological signal acquisition system. Blood rheology indicators and blood biochemical indicators were detected by Tianjin Dean Diagnostic Laboratory. Serum endothelin-1 (ET-1), homocysteine (HCY), von Willebrand factor (vWF), 8-hydroxydeoxyguanosine (8-OhdG), interleukin (IL-6) and tumor necrosis factor alpha (TNF-α) were detected by enzyme-linked immunosorbent assay (ELISA) microplate assay. Oxidative stress indicators superoxide dismutase (SOD) activity and malondialdehyde (MDA) were determined by xanthine oxidase method, thiobarbituric acid (TBA) method, reduced glutathione (GSH) and nitric oxide (NO) were tested by using microplate colorimetry. Paraffin sections were prepared from thoracic aorta tissue, and vascular structure was observed by HE staining. Results: Acute exposure to 0.12 ppm ozone could cause a significant increase in arterial systolic blood pressure (SBP). Exposure to different concentrations of ozone could cause a significant increase in plasma viscosity, and the K value of the ESR equation was significantly increased in the 1.0 ppm ozone exposure group. Both the relative and reduced viscosities were significantly reduced at ozone concentrations of 0.5 ppm and 4.0 ppm, while the red blood cell deformation index was increased significantly at ozone concentrations of 0.12 ppm, 0.5 ppm, 1.0 ppm, and 2.0 ppm. Acute ozone exposure resulted in the decrease of total cholesterol content. The content of high-density lipoprotein cholesterol (HDL-C) was significantly reduced in the 0.12 ppm ozone exposure group. When the ozone concentration was higher than 1.0 ppm, the body may also had an inflammatory reaction (increased TNF-α) and oxidative stress (increased MDA, decreased GSH). Acute exposure to ozone could lead to elevated levels of ET-1 in the blood, with significant differences in the 4.0 ppm concentration group, while HCY levels were decreased firstly and then increased, reaching the highest in the 1.0 ppm concentration group. No obvious pathological changes were observed in the thoracic aorta. Conclusion: Acute ozone exposure can affect arterial blood pressure, blood rheology and cholesterol metabolism in rats. The possible mechanism is that ozone exposure leads to inflammatory reaction and oxidative stress reaction, causing vascular endothelial function damage, and vascular endothelial cells increase with ozone exposure concentration.
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Current status of blood pressure control in patients with coronary heart disease
ZHANG Yue, WANG Tian-tian, GUO Shu-li, HAN Li-na, ZHANG Li, LI Xiao-ying, FAN Jin
CJAP    2018, 34 (1): 23-27.   DOI: 10.12047/j.cjap.5599.2018.007
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Objective: To evaluate the level of blood pressure control in patients with coronary heart disease (CHD) of China in order to provide guidance for the prevention and treatment of CHD.Methods: The patients with CHD were retrospectively collected from 2011~2014 in PLA General Hospital and Hainan Branch Hospital. Then analyzed the difference of blood pressure compliance rate between different surgical methods percutaneous coronary intervention (PCI), coronary artery bypass grafting(CABG), secondary preventive drugs(aspirin, clopidogrel, nitrates, trimetazidine, nicorandil, hypotensor, hypoglycemic, lipid-lowering drugs) and lifestyle(smoking, drinking, exercise).Results: ①Effects of surgical methods on blood pressure:Male's systolic blood pressure (SBP) and diastolic blood pressure(DBP) in the CABG group were lower in the PCI group and control group, and female's DBP in the CABG group were lower in the PCI group. ②Usage rate of secondary prevention drugs:usage rate of trimetazidine, calcium antagonist, β-blockers, angiotensin-converting enzyme inhibitor (ACEI) and angiotensin receptor antagonist (ARB) in hypertension group were higher than in normal blood pressure group. ③ Lifestyle condition:compliance rate of blood pressure in the smoking group was lower than that in the non smoking group. And there was no significant difference in blood pressure compliance rate among whether drinking and doing exercise or not.Conclusion: Blood pressure control in patients with CHD was still not satisfied. Compared with PCI, CABG may be more beneficial in the control of blood pressure in patients with CHD. Smoking cessation and improving the usage rate of secondary preventive drugs are still the main means of blood pressure control.
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Effects of chloride channels on hemolysis induced by puerarin injection
JIANG Feng-qi, YUAN Ying, XIANG Zuo-jun, HUANG Pan-rui, WU He, WANG Shi-qi, WANG Juan, PENG Hui, LUO Hai
CJAP    2018, 34 (5): 441-444.   DOI: 10.12047/j.cjap.5681.2018.100
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Objective: To observe the effects of blocking and activating chloride channels on hemolysis induced by puerarin injection in rabbits and to investigate the roles of chloride channels in hemolytic reaction induced by puerarin injection. Methods: Rabbit erythrocyte suspension was incubated with different concentrations of puerarin injection(0.75, 1.5, 3, 6, 12 mg/ml) at 37oC for 6 hours. The cell imaging system was employed to observe whether puerarin injection induced hemolysis. The hemolysis rate was detected by microplate reader and flow cytometry. Effects of activating and closing chloride channels on the hemolysis induced by puerarin injection were explored. Results: Puerarin injection could induce the hemolysis of rabbit erythrocytes in vitro. In the range of 1.5 mg/ml~12 mg/ml, puerarin injection could induce hemolysis in a concentration-dependent manner (n=3, P<0.01). The chloride channel blockers tamoxifen (20 μmol/L) and ATP (10 mmol/L) significantly inhibited the hemolysis induced by puerarin injection (n=3~5, P<0.01). Application of low concentration ATP (50 μmol/L) to activate the chloride channel significantly increased puerarin injection induced hemolysis (n=4, P<0.01). Conclusion: The hemolytic effect of puerarin injection is dose-dependent in vitro, and the activation of chloride channel is closely related to the hemolysis induced by puerarin injection.
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Effects of astragaloside-IV on the expression of inflammatory factor and proliferation of glomerular mesangial cells induced by angiotensin Ⅱ
XIONG Li, LYU Meng-juan, DOU De-yu, MA Yu-hong
CJAP    2018, 34 (5): 414-417.   DOI: 10.12047/j.cjap.5660.2018.094
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Objective: To study the effects of astragaloside-IV (As-IV) on the expression of inflammatory factor and proliferation of glomerular mesangial cells (GMCs) induced by angiotensin Ⅱ(AngⅡ). Methods: The in vitro model of diabetic nephropathy(DN) was mimic by angiotensin Ⅱ (10-6mol/L)inducing GMCs injury. Then the GMCs were treated with As-IV at different concentrations(25,50,100 μmol/L)for 48 hours. The proliferation of GMCs was detected by MTT. The level of reactive oxidative species (ROS) was measured by flow cytometry. The expression of monocyte chemoattractant protein-1(MCP-1) protein in supernatant was detected by enzyme- linked immunosorbent assay (ELISA). The expression of transforming growth factor-β1(TGF-β1) in GMCs was measured by Western blot. Results: Compared with model group, the proliferation of GMCs was inhibited in As-IV group. As-IV decreased the level of intercellular ROS, down-regulated the secretion of MCP-1 and the expression of TGF-β1 proteins. Conclusion: As-IV could inhibit cell proliferation and inflammatory factors expression on GMCs induced by AngⅡ.
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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
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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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The exploration of aerobic power and energy expenditure of Chinese rugby players
FU Tao, YANG Yue-wen, WU Peng, LIU Guan-jun
CJAP    2019, 35 (3): 215-218.   DOI: 10.12047/j.cjap.5760.2019.046
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Objective: To explore aerobic power and energy expenditure of high level rugby players in China, which provide experimental basis for accurate training and nutritional strategy in match-play. Methods: Eighteen master rugby players were selected as research subjects. The parameters such as VO2max, lactic aicd threshold (LT) and modify conconi test were measured respectively. The differences of energy were compared between the forward and the defender. The data were analyzed by independent sample t test. Results: The VO2max(42.05±3.69 ml/min·kg-1) of rugby players was poorer. The VO2max of the forward was 38.83±3.52 (ml/min·kg-1), and that of the defender was 47.31±3.17 (ml/min·kg-1),and there was significant difference between the forwards and the defenders (P<0.05). The LT of the defenders was obviously higher than that of the forwards. Modifier conconi test had a high correlation (r = 0.772) with VO2max. The average energy consumption in the first half of the game was about(276.94±18.08)kcals, the second half was(225.58±22.86)kcals, and the second half was less than the first half (P<0.05).Conclusion: The aerobic power is different between the forwards and the defenders. The power of aerobic of Chinese players is weaker than that of the foreign rugby players.
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Bioinformatics analysis of genes related to poor prognosis of human hepatocellular carcinoma and its clinical significance
XI Yi-bo, ZHANG Hao-min, YANG Bo, CHEN Xi-meng, HE Pei-feng, LU Xue-chun
CJAP    2019, 35 (1): 90-96.   DOI: 10.12047/j.cjap.5764.2019.021
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Objective:To screen genes associated with poor prognosis of hepatocellular carcinoma (HCC) and to explore the clinical significance of these genes. Methods: The proper expression profile data of HCC was obtained from the Gene Expression Omnibus (GEO) database, and the differentially expressed genes (DEGs) were identified by differential expression analysis. The DAVID and String database were used for function enrichment analysis and to construct the protein-protein interaction (PPI) network respectively. The Cancer Genome Atlas (TCGA) database and the Cox Proportional Hazard Model were used for prognosis analysis of the DEGs. Results: A eligible human HCC data set (GSE84402) met the requirements. A total of 1141 differentially expressed genes were identified, including 720 up-regulated and 421 down-regulated genes. The results of function enrichment analysis and PPI network performed that CDK1、CDC6、CCNA2、CHEK1、CENPE 、PIK3R1、RACGAP1、BIRC5、KIF11 and CYP2B6 were prognosis key genes. And the prognosis analysis showed that the expressions of CDC6、PIK3R1、KIF11 and RACGAP1 were increased, and the expression of CENPE was decreased, which was closely related to prognosis of HCC. Conclusion: CDC6、CENPE、PIK3R1、KIF11 and RACGAP1 may be closely related to poor prognosis of HCC, and can be used as molecular biomarkers for future research of HCC prognosis.
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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
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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 effects of anthocyanin on chronic inflammatory pain induced by complete Freund's adjuvant and its mechanism
WANG Jing, CHEN Ze-feng, HE Chun-xia, WANG Li-li, ZHENG Jia-hui, ZHANG Hao, WANG Zhan-lei, LU Ya-ping
CJAP    2018, 34 (5): 476-480.   DOI: 10.12047/j.cjap.5671.2018.106
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Objective: To investigate the effects and mechanisms of anthocyanin from Ligustrum vicaryi on chronic inflammatory pain induced by complete Freund's adjuvant. Methods: Thirty male SD rats were randomly divided into three groups (n=10):normal saline control group (NS), chronic inflammatory pain model group(Mod, injected with complete Freund's adjuvant(CFA) 100 μl to the left hind leg), anthocyanin treatment group(Ant, dosed with anthocyanins (90 m), mechanical pain threshold (MPT), and left toe volume in each group were measured before modeling and 1,3,5,7,9,11,13 days after operation. Antioxidant indexes in serum were mensurated by spectrophotometer, and the total capsaicin receptor (TRPV1) and phosphorylated capsaicin receptor (p-TRPV1) in hippocampus were detected by Western blot. Results: In comparison with controls, HPT and MPT were improved (P<0.05),toe swelling was reduced(P<0.05), the serum level of SOD was increased (P<0.01), while the levels of MDA and NO were decreased (P<0.05), the ratio of P-TRPV1/TRPV1 protein was depressed in Mod rat hippocampal region treated with anthocyanin. Conclusion: The results show that anthocyanins has an analgesic effect on chronic inflammatory pain induced by CFA, and its mechanism may be related to the improvement of antioxidant capacity and the reduction of TRPV1 phosphorylation.
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