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  • Table of Content
      28 March 2018, Volume 34 Issue 2 Previous Issue    Next Issue
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    Changes in expression of autophagy-related factors during acute contusion repair of skeletal muscle
    LUO Ao, TANG Cheng-lin, HUANG Si-qin, ZHAO Dan-dan, ZHANG An-ning, GUO Quan-hu, GAO Rui-qi, CAO Jing
    CJAP. 2018, 34 (2): 97-101.   DOI: 10.12047/j.cjap.5647.2018.024
    Abstract   PDF (1160KB) ( 145 )
    Objective:To explore the possible biological mechanism of skeletal muscle contusion repair through researching the changes in expression of autophagy-related genes and proteins in SD rats with acute skeletal muscle contusion. Methods:Six rats were randomly selected as the control group from 30 male SD rats, acute skeletal muscle contusion model were established in the remaining 24 rats with self-made hitter, then the model rats were randomly divided into 4 groups (3 d, 5 d, 7 d, 14 d groups, n=6). On the 3rd, 5th, 7th and 14th day after injury, injured gastrocnemius of each group was harvested. The morphological and the ultra-microstructure changes of gastrocnemius after injury were observed by HE staining and transmission electron microscope (TEM) respectively. The relative protein levels of (LC3-Ⅱ) and P62 of each group were observed by Western blot. The relative mRNA levels of atg7, atg10, atg12, atg16L1 of each group were observed by RTPCR. Results:The results of HE staining showed that compared with the control group, the inflammation reached its peak on the 5th day after injury, new muscle fibers were clearly observed in 7 d group. The results of TEM showed that, compared with the control group, oncotic mitochondria could be clearly seen in the 3 d, 5 d, 7 d groups. Also, the Z line changed from disappearing to drift thickening, sarcoplasmic reticulum dilatation gradually improved, there was no evident difference between the 14 d group and the control group, suggesting that the damage has preliminarily healed. The results of Western blot showed that the expressions of LC3-Ⅱand P62 were increased at first and then decreased. The expression of LC3-Ⅱwas markedly up-regulated in the 3 d, 5 d, 7 d groups compared with the control group and the 14 d group (P<0.01). Similarly, compared with the control group, the expression of P62 reached its peak on the 3rd day after injury (P<0. 01), and returned to normal level on the 14th day. The results of RT-PCR showed that the expression of atg10 mRNA in the natural recovery group of 3 d, 5 d, 7 d, 14 d was firstly decreased and then increased, the atg10 mRNA was markedly down-regulated in the 3 d, 5 d, 7 d groups compared with the control group and the 14 d group (P<0. 01). The expression of atg7, atg12, atg16L1 mRNA was generally increased at first and then decreased, it was markedly up-regulated in the 3 d, 5 d, 7 d groups compared with the control group and the 14 d group (P<0.01, P<0.05, P<0.01). Conclusion:The above results indicate that the autophagy is involved in repair of skeletal muscle injury by its autophagyrelated factors,regularly changes after contusion, and the rate of damage repair may be related to the level of autophagy.
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    Effect of autophagy inhibitor chloroquine on acute alcoholinduced liver disease
    SANG Wen-hua, ZENG Mei-Chun, CHEN Sha, CHEN Ran, FAN Xiao-fang, GONG Yong-sheng, ZHANG Hai-lin, ZHANG Hong-yu, KONG Xiao-xia
    CJAP. 2018, 34 (2): 102-105.   DOI: 10.12047/j.cjap.5551.2018.025
    Abstract   PDF (939KB) ( 57 )
    Objective:To investigate the role of autophagy inhibitor chloroquine (CQ) in acute ethanol-induced liver injury and its mechenism. Methods:Twenty-one C57BL/6 male mice were randomly divided into three groups:control group, ethanol group, CQ + ethanol group (n=7). Mice in ethanol group were administered 33% (v/v) ethanol at a dose of 4.5 g/kg body weight. Ethanol-induced liver steatosis in each group was detected by hematoxylin and eosin staining. Hepatic lipid accumulation was detected by staining with Oil red O. Hepatic tissue triglyceride (TG) levels, serum aspartate aminotransferase(AST) and alanine aminotransferase(ALT) were determined by biochemical assays. Protein expression of microtubule-associated protein 1 light chain 3(LC3) and nuclear factorκB p65(NF-κB p65) were measured by Western blot and immunofluorescence. Pro-inflammatory factors tumor necrosis factor-α(TNF-α)、interleukin 6(IL-6) were detected by ELISA. Results:Compared with control group, ethanol induced liver injury proved by accumulation of hepatic lipids, TG levels, AST and ALT activities were significantly increased by ethanol, protein expression of LC3-Ⅱ was also markedly increased by ethanol. Compared with ethanol group, addition of CQ increased furtherthe level of LC3-Ⅱexpression, and TG amount, serum AST and ALT activities, and the expression of NF-κB p65, TNF-αand IL-6. Conclusion:Acute ethanol-intake could induce liver steatosis and inflammation, and autophagy inhibitor CQ exacerbatedethanol-induced liver injury, suggested that autophagy might be protective effect in acute ethanol-induced liver disease.
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    Anti-arrhythmic effects of taurine-magnesium coordination compound on torsades de pointes
    LI Yan, SUN Kai, AN Meng-yao, PAN Ying-ying, SUN Tao, YIN Yong-qiang, KANG Yi, LOU Jian-shi
    CJAP. 2018, 34 (2): 106-110.   DOI: 10.12047/j.cjap.5596.2018.026
    Abstract   PDF (1489KB) ( 74 )
    Objective:To investigate the effect of taurine magnesium coordination compound (TMCC) on torsades de pointes (TdP) in isolated guinea pig hearts. Methods:Healthy male guinea pigs weighting 250~300 g were randomly divided into 4 groups:①TdP model group (n=7):Isolated hearts were perfused by normal K-H solution 20 minutes, then perfused by slowly activated delayed rectifier potassium current(IKs) blocker 10μmol/L Chromanol 293B under hypokalemic solution(1.8 mmol/L) to establish TdP model;②~④ TdP model + TMCC group (n=6):Isolated hearts were perfused by normal K-H solution for 20 minutes, then perfused by IKs blocker 10μmol/L Chromanol 293B under hypokalemic solution(1.8 mmol/L) for 60 minutes, at the same time TMCC which concentration was 1, 2, 4 mmol/L was administered respectively by Langendorff retrograde aortic perfusion method. Cardiac surface electrocardiogram of guinea pigs in vitro was collected and recorded by Biopac electrophysiological recorder. Incidence of TdP, transmural dispersion of repolarization (TDR), instability of QT interval were acquired from Lead Ⅱ electrocardiograph (ECG) wave forms to describe the effect of TMCC on TdP model. Datas were acquired at the time of 20 min and pre-TdP, in case there was no TdP observed, a value of 60 min was entered for calculation purpose. Results:Incidence of TdP in TdP model group was 6/7. TdP incidence could be decreased significantly by 1, 2, 4 mmol/L TMCC, and was 5/6, 1/6, 0/6 respectively. Compared with the pre-drug, Chromanol 293B under hypokalemic solution in TdP model group increased TDR(corrected) evidently(P<0.01). Compared with the pre-drug, 1, 2, 4 mmol/L TMCC in TdP model + TMCC group could decrease the increased TDR(corrected) induced by Chromanol 293B under hypokalemic solution(P>0.05). Compared with the TdP model group, 2, 4 mmol/L TMCC could evidently decrease the instability of QT interval induced by Chromanol 293B under hypokalemic solution(P<0.05). During the establishment of TdP model, P waves in more than one cardiac cycle continuously were disappeared in ECG. However, P wave could always be seen independent in ECG acquired from TdP model + TMCC group. Conclusion:TMCC can play the role against TdP through decreasing TDR and instability of QT interval, and inhibiting early after depolarization(EAD).
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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
    Abstract   PDF (1106KB) ( 435 )
    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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    Effect of fracture of lower limbs with hemorrhage on myocardial injury and its mechanism in rats
    LI Zhao-hui, ZHANG Ying, LU Yan, LU Qing-ming, XIE Xiao-hua
    CJAP. 2018, 34 (2): 115-119.   DOI: 10.12047/j.cjap.5558.2018.028
    Abstract   PDF (1246KB) ( 54 )
    Objective:To test whether myocardial apoptosis can be induced by traumatic fracture of lower limbs with hemorrhage, in order to lay a foundation of myocardial injury after traumatic fracture for the follow-up study. Methods:Twenty SD rats were randomly divided into two groups, i. e. control group and trauma group(n=10). A rat model of traumatic hemorrhage was establish, and a traumatic model of the original generation of myocardial cell culture was constructed in vitro. The level of interleukin-2(IL-2),IL-6,IL-10 and tumor necrosis factor-α(TNF-α) in rat serum was detected by ELISA at 0, 1, 2, 4, 8, 12, 16, 24 and 48 hour to find the most significant point. The pathological cardiac injury in rats was observed by HE staining under a microscope, and the apoptosis of cultured cardiomyocyte in vitro was detected by TUNEL methods. The expressions of apoptosis gene,(Bcl-2) and Bax, in myocardium of rat and cultured cardiomyocyte in vitro were detected by Western blot and RT-PCR. Results:At the 4th hour after trauma, IL-6 and IL-10 in the serum of rats reached its highest, IL-2 reached its lowest at the 8th hour after trauma, and TNF-αreached its highest at 1 hour after trauma, then all recovered to their normol level gradually. Myocardial HE staining indicated that cardiomyocyte was swelling, disordered derangement, inflammatory cell infiltrated; a large number of myocardial cell nuclei was dyedbrown in TUNEL test which proved that the apoptosis index increased (P<0.05). Western blot and RT-PCR results showed that the expression of pro-apoptotic gene Bax was up-regulated (P<0. 05), while expression of anti apoptosis gene Bcl-2 down-regulated (P<0.05). Conclusion:The myocardial apoptosis can be induced by traumatic fracture of lower limbs with hemorrhage in rats, and then lead to myocardial injury.
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    Effects of centellaasiatica granule on the expression of Smad 2/3, Smad 7 and collagen Ⅳ in the mesangial cells stably expressed TGF-β1
    MA Ji-wei, WANG Hong-tian, LIU Hao-fei, DONG Lei-peng, DING Yuan, BAI Ji-qiong, ZHANG Zhu, DONG Li-jie
    CJAP. 2018, 34 (2): 122-125.   DOI: 10.12047/j.cjap.5568.2018.030
    Abstract   PDF (1455KB) ( 127 )
    Objective:Stably expressed transforming growth factor -beta 1(TGF-β1)MCs were obtained and the effects of centellaasiatica (CA) granule on the expressions of Smad 2/3, Smad 7 and collagen Ⅳ and the level of Smad 2/3 phosphorylation were observed. Methods:Lipofectin method was used to transfect TGF-β1 vector into MC, and the stably expressed TGF-β1 cell lines were selected by G418. The cells were divided into three groups. Control group:normal MC + RPMI 1640 + 10% normal rat serum; TGF-β1 group:stably expressed TGF-β1 MC + RPMI 1640 + 10% normal rat serum; CA group:stably expressed TGF-β1 MC + RPMI 1640 + 10% rat serum containing high CA. The experiments were repeated for five times. The contents of TGF-β1 and collagen Ⅳ in the culture medium were detected with ELISA, the expressions of mRNA and protein of TGF-β1, Smad 2/3, Smad 7 and the level of Smad 2/3 phosphorylation were detected by using real time quantitative polymerase chain reaction and Western blot. Results:The contents of TGF-β1 and collagen Ⅳ in the culture medium of stably-expressed TGF-β1 MC were increased significantly, and the CA could reverse the effects of TGF-β1. The expressions of mRNA and protein of TGF-β1, Smad 2/3 and the level of Smad 2/3 phosphorylation were increased significantly in TGF-β1 transfected MC, and CA could dramatically reduce the expressions of mRNA and protein of TGF-β1, Smad 2/3 and the level of Smad 2/3 phosphorylation. The high expression of TGF-β1 decreased the expression of Smad 7 mRNA and protein, and the CA could antagonize the effect of mRNA expression. Conclusion:The MCs stably-expressed TGF-β1 can activate the TGF-β1/Smad signal pathway and increase the expression of collagen Ⅳ. CA can decrease the occurrence of diabetic nephropathy(DN) by reducing the production of collagen Ⅳ through inhibiting the TGF-β1/Smad signal pathway.
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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
    Abstract   PDF (929KB) ( 110 )
    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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    Effect of two kinds of fermented food on blood glucose and lipids in induced diabetic mice
    YAN Yong-qiu, DAI Guan-hai, JIANG Shi-ping, LU Yu, TONG Ying-peng, WANG Ping
    CJAP. 2018, 34 (2): 130-133.   DOI: 10.12047/j.cjap.5547.2018.032
    Abstract   PDF (937KB) ( 118 )
    Objective:To investigate the effect of two kinds of fermented food, noni juice and natto, on blood glucose and lipids in induced diabetic mice. Methods:Female (ICR) mice were induced into diabetes by an injection of alloxan (55 mg/kg, i. v.). After 72 hours, those mice whose fasting blood glucose levels were over 12.00 mmol/L and urine sugar was strongly positive (+ + +) were regarded as diabetical model and were randomly divided into three groups (n=10):diabetical model (DM) group, noni juice (NJ) group and natto (NT) group. Another ten normal female ICR mice were taken as normal control (NC) group. The mice in NJ and NT groups were gavaged with noni juice (25.0 ml/kg) and natto (0.6 g/kg) respectively. The other two groups were given normal saline (25.0 ml/kg). Continuous gavage administration was given for 30 days, the water-drinking volume and food-intake were recorded. After 1.5 h of the last administration, the glucose tolerance of mice was measured. Finally, the changes in glycated serum protein(GSP), insulin(Ins) and blood lipids of blood samples of mice, taken from the femoral artery, were determined. Results:Compared to the NC group, the water-drinking amount and food-intake, GSP and total cholesterol (TC), triglyceride(TG) and low density lipoprotein (LDL) in DM group were increased significantly (P<0.01), while glucose tolerance, Ins and high density lipoprotein (HDL) were decreased significantly (P<0.01). However, when it came to DM group, NJ and NT could significantly (P<0.01 or P<0.05) reduce, GSP, TG and LDL, meanwhile improve glucose tolerance, Ins and HDL (P<0. 01). Conclusion:Both noni juice and natto could reduce the blood glucose levels in induced diabetical mice and improve blood lipids, which suggested that they may have certain application value in prevention and treatment of diabetic mellitus.
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    Effects of ursolic acid on liver injury and its possible mechanism in diabetes mellitus mice
    WANG Xu-tao, CHEN Si-si, QI Min-you
    CJAP. 2018, 34 (2): 134-136.   DOI: 10.12047/j.cjap.5598.2018.033
    Abstract   PDF (1007KB) ( 142 )
    Objective:To study the effects of ursolic acid on liver injury in diabetic mice induced by high-fat diet combined with streptozotocin(STZ), and to explore its possible mechanisms. Methods:Diabetes mellitus was induced in twenty male ICR mice by a combination of high-fat diet for 6 weeks with low-dose streptozotocin (30 mg/kg, i. p.) for 5 consecutive days. After 9 days, fasting blood glucose levels were determined. Mice with fasting blood glucose levels exceeded 11. 1 mmol/L were diagnosed as diabetic mice and selected for further experiment. These mice were randomly divided into two groups(each group of 10):diabetic group, ursolic acid group (100 mg/kg, i. g.), and another 10 mice were set as control group. After continuous administration for 8 weeks, body weight (BW) were weighed, fasting blood glucose (FBG), total cholesterol (TC), triglyceride (TG), alanine aminotransferase (ALT), aspartate transaminase (AST) in serum and superoxide dismutase (SOD), malondialdehyde (MDA) in liver were measured. HE staining was used to observe pathological changes of liver tissue. Results:Compared with the control group, the level of FBG, TC, TG, ALT, AST, MDA were dramatically increased (P<0. 05, P<0. 01) and SOD was markedly decreased (P<0.01) in the diabetic group; HE staining showed that parts of liver cells swelled and had a light fatty degeneration as well as lymphocyte infiltrated around the portal area in model group. Compared with the diabetic group, the level of FBG, TC, TG, ALT, AST, MDA were significantly declined (P<0.05, P<0.01) and SOD was considerably increased (P<0.01) in the ursolic acid group; HE staining showed that the liver cells relatively arranged in order, edema was not obvious and inflammatory cells infiltrated lightly in the ursolic acid group. Conclusion:Ursolic acid has a protective effect on liver injury in diabetic mice induced by high-fat diet combined with STZ by intraperitoneal ingector, and its mechanism may be associated with lowering blood glucose, regulating the lipid metabolism, reducing oxidative stress and enhancing the ability of anti-oxidation in liver.
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    Effect of dexmedetomidine on apoptosis and CHOP in hypoxia/reoxygenation injury A549 cell
    LUO Zi-yin, GAO Hui, XIANG Bing-qian, QIU Xiao-xiao, DAI Yong-yue, WANG Wan-tie
    CJAP. 2018, 34 (2): 137-142.   DOI: 10.12047/j.cjap.5554.2018.034
    Abstract   PDF (1672KB) ( 21 )
    Objective:To investigate the effects of dexmedetomidine (Dex) on injury of A549 cells induced by hypoxia/reoxygenation(H/R)and the influence of C/EBP homologous protein (CHOP) expression. Methods:Logarithmic growth phase A549 cells(it originated from alveolar type Ⅱ epithelial cell line) were randomly divided into 4 groups (n=10):normoxic control group (N), Dex group (D), hypoxia/reoxygenation group (H), hypoxia/reoxygenation + Dex group(HD). At the beginning of modeling, 1 nmol/L Dex was puted into D and HD groups. N and D groups were cultured in the normoxic incubator for 30 h. H and HD group were incubated in the anoxic cultivation for 6 h, fo llowed by normoxic culture for 24 h. Then A549 cells were observed under the inverted microscope to observe the morphological changes. Cell activity was detected by cell counting Kit-8(CCK-8) and the apoptosis index(AI) was detected by in situ end labeling (TUNEL) method. The expression of CHOP、glucose-regulated protein of molecular weight 78 kDa (Grp78)、cysteinyl aspirate-specificprotease-3 (caspase-3) protein and CHOP、Grp78 mRNA were detected by Western blot and RT-PCR. Results:Compared with N group, the number of adherent cells in H group decreased significantly, and cell morphology changed. The absorbance value in H group decreased obviously (P<0. 01). The AI value and expression of CHOP, Grp78, caspase-3 proteins and CHOP, Grp78 mRNA were significantly increased (P<0.01). Compared with H group, the cell damage in HD group was decreased, the absorbance value increased (P<0.01), the number of apoptosis cells decreased relatively (P<0.01), the expression of CHOP, caspase-3 protein and CHOP mRNA decreased (P<0. 01). Conclusion:Dex has notable effects against H/R injury, which may be related to effective inhibition of apoptosis mediated by the CHOP's signal path.
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    Effects of 16 week aerobic exercise on baPWV and ABI of middle-aged and elderly patients
    LI Ning-chuan, YIN Xia-lian, WEI Xiu-xia, LU Wei, SHEN Shi-wen, XIAO Lang, HU Yu-long
    CJAP. 2018, 34 (2): 145-149.   DOI: 10.12047/j.cjap.5577.2018.036
    Abstract   PDF (950KB) ( 140 )
    Objective:To investigate the interventional effects of 16-week aerobic exercises on the elderly's arteriosclerosis and its mechanism. Methods:Twenty-seven elderly people with the average age of 62. 70 ±3. 26 joined a 16-week square dance/taijiquan exercise program that conducted 60 minutes each time, six times per week. Arterial stiffness and its related indexes such as systolic pressure(SBP), diastolic pressure(DBP), left brachial-ankle pulse wave velocity (L-baPWV), right brachial-ankle pulse wave velocity(R-baPWV), left ankle brachial index (L-ABI), right ankle brachial index(R-ABI), serum triglyceride(TG), total cholesterol(TC), high density lipoprotein cholesterol(HDL-c), low density lipoprotein cholesterol(LDL-c), superoxide dismutase(SOD), malondialdehyde(MDA) and glutathione peroxidase (GSH-Px) were detected at 3 time points including before exercise program, by the end of exercise for 8 weeks and 16 weeks. Results:① Compared with pre-exercise, the R-baPWV and R-ABI of the elderly people were decreased at the end of the 8th week, and the L-baPWV, RbaPWV, R-ABI and L-ABI were decreased significantly at the end of the 16th week. ②Compared with pre-exercise, SBP and DBP were declined markedly (P<0.01, P<0.05) at the end of the 8th week, SBP, DBP and pulse pressure were decreased significantly (P<0.01, P<0.05) at the end of the 16th week. ③Compared with pre-exercise, TC and LDL-c were declined markedly (P<0.01) at the end of the 8th and the 16th week, and there was no difference of the level of TG and LDL-c between pre-exercise and post-exercise. ④There was no evident difference of serum level of SOD, GSH-Px, MDA between pre-exercise and post-exercise at the end of the 8th week. Compared with pre-exercise, the level of serum SOD, GSH-Px was increased evidently while the content of serum MDA was decreased significantly (P<0.01). Conclusion:Sixteen-week aerobic exercises could reduce baPWV and ABI levels, regulate blood pressure, blood lipids and lipid peroxides levels of the elderly evidently, thus improve the controlling quality of atherosclerosis.
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    Effects of tetramethylypyrazine nitrone on proliferation and differentiation of neural stem cells in vitro
    CHEN Shi-jin, CHEN Dong, SHI Yu-fang, LIU jun, HAN Song, LI wei
    CJAP. 2018, 34 (2): 150-153.   DOI: 10.12047/j.cjap.5543.2018.037
    Abstract   PDF (1129KB) ( 32 )
    Objective:To investigate the role of tetramethylpyrazine(TMP) nitrone in proliferation and differentiation of neural stem cells (NSCs). Methods:We separated and cultivated the original generation of NSCs from cerebral cortex of 14 days rat embryo, and the phenotype characteristics of the third-generation NSCs was tested by immunofluorescence. The experiment was divided into control group, β-mercaptoethanol positive control group, tetramethylpyrazine nitrone group and tetramethylpyrazine nitrone + ethylene glycol tetraacetic acid(EGTA) group (n=4). The third-generation cultivation of NSCs was used in the experiment. The effect of tetramethylpyrazine nitrone on the number of NSCs proliferation was determined by BrdU and MTT, and the differentiation of NSCs was determined by Western blot. Results:The primary NSCs was isolated successfully, neurospheres with typical NSCs morphology and expressing nestin was formed at 3-5 days. As BrdU and MTT assay results shown, compared with the control group andβ-mercaptoethanol positive control group, the NSCs proliferation numbers of tetramethylpyrazine nitrone group increased significantly(P<0.05). The results of Western blot showed that the neuronal differentiation rate of NSCs was increased significantly in both the tetramethylpyrazine nitrone group and tetramethylpyrazine nitrone + EGTA group, and the differentiation rate of NSCs in tetramethylpyrazine nitrone + EGTA group increased more significantly(P<0.05). Conclusion:Tetramethylpyrazine nitrone can significantly enhance the proliferation and neuronal differentiation rate of NSCs. Decrease in extracellular Ca2+ can promote the differentiation of NSCs into neurons induced by tetramethylpyrazine nitrone. Ca2+ signaling plays an important role in the differentiation of NSCs into neurons.
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    The influence of benazepril and amlodipine on the expression of secretin and somatostatin in spontaneously hypertensive rats
    JIN Hua, LIU Zhi-jun, YAN Chun-lu, LIU Feng-lin, CHEN Li, ZHANG Qiu-ju, XU Hou-qian, HU Ji-hong, DOU Rong-hai, WEN Xin-yang
    CJAP. 2018, 34 (2): 154-158.   DOI: 10.12047/j.cjap.5542.2018.038
    Abstract   PDF (950KB) ( 35 )
    Objective:Investigate the influence of benazepril and amlodipine on the expression of secretin (PZ) and somatostatin (SS) in spontaneously hypertensive rats (SHR). Methods:Forty-five SHRs (14 weeks old, male) were randomly assigned into 3 groups (n=15):SHR group, Benazepril group (which was given benazepril 0.90 mg·kg-1·d-1) and Amlodipine group (SHRs were given amlodipine 0.45 mg· kg-1·d-1), taking WistarKyoto(WKY) as normal control (n=15), meanwhile, rats in SHR group and WKY group were given the same volume of distilled water. After 8 weeks of intervention, the expression of protein and mRNA of PZ in duodenum and SS in sinuses ventriculi was detected by enzyme-linked immunoassay and RT-PCR. Results:After 8 weeks of intervention, compared with the WKY group, the expression of protein and mRNA of PZ in duodenum and SS in sinuses ventriculi was increased significantly in SHR group (P<0. 05). Compared with SHR group, the expression of PZ in duodenum and SS in sinuses ventriculi was decreased significantly in Benazepril group and Amlodipine group (P<0.05). Compared with Benazepril group, in Amlodipine group the expression of PZ mRNA in duodenum and SS mRNA in sinuses ventriculi was decreased more significantly (P<0.05). Conclusion:The regulation disorder of PZ in duodenum and SS in sinuses ventriculi exists in SHR. The antihypertensive effect of benazepril and amlodipine may be realized by regulating the expression of PZ and SS, while the regulation of amlodipine is more obvious than benazepril.
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    Effect of Astragalus injection on cardiomyocyte apoptosis, endoplasmic reticulum stress and expression of connexin in cardiomyopathy rats induced by adriamycin
    HE Zhi-hui, SHAO Li-qun, XUAN Li-ying, WANG Chun-gui, WEI Cheng-xi, WANG Yu, ZHAO Ming
    CJAP. 2018, 34 (2): 159-163.   DOI: 10.12047/j.cjap.5576.2018.039
    Abstract   PDF (1862KB) ( 142 )
    Objective:To investigate the effect of Astragalus injection on cardiomyocyte apoptosis, endoplasmic reticulum stress and connexin protein in cardiomyopathy rats induced by adriamycin. Methods:Thirty-six male Wister rats were randomly divided into control group (n=12), adriamycin(ADR) group (n=12) and Astragalus group (n=12). The normal saline (10 ml/kg body weight) was injected intraperitoneally in control group rats, ADR (2 mg/kg body weight) was injected intraperitoneally in ADR group rats, ADR (10 ml/kg body weight) and Astragalus injection (10 ml/kg body weight) were injected intraperitoneally in rats of astragalus group, one time a week, totle 3 times. By the end of the 7th week, the left ventricular end-diastolic diameter(LVEDD), left ventricular end-systolic diameter (LVESD) and left ventricular ejection fraction (LVEF) were measured by echocardiography. Then the rats in the three groups were sacrificed and the left ventricle section was stained by HE, Masson, uranyl acetate/lead citrate respectively, the cardiomyopathy and ultrastructural changes were observed under light microscope and transmission electron microscope. The apoptosis of rat cardiomyocyte were analyzed by TUNEL. The expression of connexin Cx43 and p-Cx43 was detected by immunohistochemistry. The expression of glucose-regulated protein 78 (Grp78),activating transcription factor 4 (ATF-4) and C/EBP homologous protein (CHOP) were detected by real time PCR. Results:Compared with control group, LVEDD, LVESD increased and LVEF decreased, myocardial fibers were disordered and edematous, infiltrated by lymphocytes, the mitochondria were destroyed and vacuolized, and the number of cardiomyocyte apoptosis was increased(P<0.01) in ADR group. The expression of Grp78, ATF-4, CHOP and p-Cx43 were increased, and the expression of Cx43 was decreased in ADR group. However, compared with ADR group, LVEDD, LVESD decreased and LVEF increased, the cardiomyopathy and ultrastructural changes were significantly improved, the number of cardiomyocyte apoptosis was significantly decreased (P<0. 01); the expression of Grp78, ATF-4, CHOP and p-Cx43 decreased (P<0.01); the expression of Cx43 increased in Astragalus group (P<0.01). Conclusion:Astragalus injection may effectively improve the myocardial damage induced by adriamycin, its mechanism may be related to the inhibition of endoplasmic reticulum stress (ERS) and the decrease of phosphorylation of CX43 in cardiomyopathy rats induced by adriamycin.
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    The protective effects of Astragaloside Ⅳ on diastolic function of rat thoracic aortic rings impaired by microvesicles
    LI Ye-yi, SHANG Man, ZHANG Kun-wei, WEI Su, LIU Chao, ZHU Qian, ZHAO Jun-yu, WU Yan-na, SONG Jun-qiu, LIU Yan-xia
    CJAP. 2018, 34 (2): 164-168.   DOI: 10.12047/j.cjap.5659.2018.040
    Abstract   PDF (1236KB) ( 63 )
    Objective:To investigate the effects of Astragaloside IV (AST) on diastolic function of rat thoracic aorta rings which was injured by microvesicles derived from hypoxia/reoxygenation (H/R)-treated human umbilical vein endothelial cells (HUVECs), and the mechanism of AST. Methods:H/R-induced endothelial microvesicles (H/R-EMVs) were generated from cultured HUVECs in vitro under the condition of hypoxia for 12 hour/Reoxygenation for 4 hour, H/R-EMVs were stored in D-Hank's solution. Male Wistar rats were underwent thoracotomy, the thoracic aorta with intact endothelium were carefully removed and cut into 3~4 mm rings. The experiment was divided into six groups. H/R-EMVs group:thoracic aortic rings of rats were incubated in culture medium and treated with H/R-EMVs in a final concentration of 10μg/ml; different doses of AST groups:thoracic aortic rings of rats were treated with 10, 20, 40, 60 mg/L AST co-incubated with 10μg/ml H/R-EMVs respectively; control group were treated with the same volume of D-Hank's solution. Duration of incubation was 4 h, each group was tested in five replicate aortic rings. Effects of AST on endothelium-dependent relaxation were detected. The production of nitric oxide (NO) and the level of endothelial NO synthase (eNOS), phosphorylated eNOS (p-eNOS, Ser-1177), serine/threonine kinase (Akt), phosphorylated Akt (p-Akt, Ser-473), extracellular regulated protein kinases (ERK1/2) and phosphorylated ERK1/2 (p-ERK1/2, Thr202/Tyr204) of rat thoracic aortic rings were detected. Results:Tenμg/ml H/R-EMVs could impaire the relaxation of rat thoracic aortic rings significantly (P<0.01). Compared with H/R-EMVs group, relaxation of rat thoracic aortic rings was increased by 20, 40 and 60 mg/L AST in a concentration-dependent manner (P<0.01), the level of NO production was also enhanced (P<0.05, P<0.01). The level of t-eNOS, t-Akt and ERK1/2 was not changed, but the level of p-eNOS, p-Akt and p-ERK1/2 increased by the treatment with AST (P<0.01). Conclusion:AST could effectively ameliorate endotheliumdependent relaxation of rat thoracic aortic rings impaired by H/R-EMVs in a concentration-dependent manner, the mechanism might involve the increase in production of NO, and the protein level of p-eNOS, p-Akt and p-ERK1/2.
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    Effect of Yuyin Ruangan Granule on TGF-β1 expression in hepatic fibrosis rats
    WANG Zhi-wang, FU Xiao-yan, CHENG Xiao-li, BAO Xiao-ying, LIU Xue-feng, DUAN Hai-jing
    CJAP. 2018, 34 (2): 169-172.   DOI: 10.12047/j.cjap.5593.2018.041
    Abstract   PDF (1202KB) ( 156 )
    Objective:To observe the preventive and therapeutic action of Yuyin Ruangan Granule (YRG, Traditional Chinese Medicine) in hepatic fibrosis rats model and its effect on transforming growth factor-β1 (TGF-β1) expression. Methods:The Wistar rats were randomly divided into 6 group (n=10), and the model of hepatic fibrosis rats was established by subcutaneous injected with carbon tetrachloride (CCL4), fed on high-fat diet and 20% ethanol for 6 weeks, to survey the effect and mechanism of YRG preventing hypatic fibrosis by detecting liver function (the activity of alanine aminotransferase(ALT), aspartate aminotransferase(AST), etc.) of liver fibrosis rats, liver fibrosis indicators (hyaluronic acid, Ⅲ procollagen, type IV collagen, laminin and hepatic pathology, etc.), and TGF-β1 expression in liver tissue after 6 weeks treated with YRG through intragastric administration (q. d.). Results:At the 7th week, fibrotic lesions appears distinctly in liver tissue of model group compared with control group (P<0.01), YRG of 6.2~28.8 g/kg could significantly decrease hepatic index, ALT and AST activities, content of hyaluronic acid(HA), Ⅲ procollagen (PCⅢ), type Ⅳ collagen(C-Ⅳ), laminin (LN) in serum, relieve liver fibrosis pathological changes and inhibit TGF-β1 expression in fibrotic liver tissue (P<0.05, P<0.01). Conclusion:YRG has significantly preventive effects on liver fibrosis rats model, and it may be one of its mechanisms to inhibit expression of TGF-β1.
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    Effect of saffron aqueous extract on the level of blood glucose in experimental diabetes mice
    JIANG Shi-ping, SHEN Qian, LU Yu, YAN Yong-qiu, TONG Ying-peng, WANG Ping
    CJAP. 2018, 34 (2): 173-176.   DOI: 10.12047/j.cjap.5588.2018.042
    Abstract   PDF (1068KB) ( 130 )
    Objective:To investigate the effects of saffron aqueous extract (SE) on blood glucose, lipid and pancreatic tissue in streptozocin-induced diabetes mice. Methods:Diabetes mellitus mice were established by intraperitoneal injection of streptozocin (60 mg/kg) for two consecutive days. The 30 well-established diabetes mice were randomly divided into three groups(n=10):diabetic mellitus (DM) group, SE treated (SE) group and positive control (metformin hydrochloride, MH) group. Another ten normal mice were selected as normal control (NC) group. The mice in SE and MH groups were intragastrically administered with SE 100 mg/kg or MH 100 mg/kg once a day for 6 weeks, mice in DM and NC were given normal saline. The amount of food-intake, water consumption and body weight were measured weekly, the changes of the indicators including fasting blood glucose (FBG), oral glucose-tolerance test (OGTT), glycated serum protein (GSP), insulin (INS) and blood lipid were determined after 6 weeks of continuous administration. The pathologic changes in the pancreas tissues were detected by HE staining. Results:Compared with the normal control group, the amount of food-intake, water consumption, area under the curve, FBG, GSP, and total cholesterol (TC) were significantly increased, while fasting weight, INS and high density lipoprotein cholesterol (HDL-c) were dramatically decreased in DM group. Compared with DM group, the water consumption, FBG, area under the curve and TC in SE group were starkly declined with a notable elevation of HDL-c and INS. In addition, the biopsy from DM mice showed the structure of pancreas islet was destroyed and reduced, and vascular proliferation with irregular shape. The damaged pancreas was obviously repaired by giving SE. Conclusion:The saffron aqueous extract had efficacy on blood glucose and blood lipids reduction, improvement on damaged pancreas in streptozocininduced diabetic mice, which suggested that saffron could be used for the treatment in diabetes.
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    The therapeutic effects of Yougui pill on knee osteoarthritis and the expression of Wnt signal pathway related factors in rats
    YAN Chun-lu, LI Sheng-hua, AN Fang-yu, LIU Yong-qi, XIA Peng-fei, MA Zheng-min, NIU Yan-qiang, LIU Peng-yao
    CJAP. 2018, 34 (2): 177-181.   DOI: 10.12047/j.cjap.5603.2018.043
    Abstract   PDF (1481KB) ( 159 )
    Objective:To observe the effects of Yougui pill (Traditional Chinese Medicine) on the related factors of Wnt signal pathway of rats with knee osteoarthritis (KOA), and explore its protective mechanism. Methods:Sixty SPF SD rats were randomly divided into the sham-operative group, model group, glucosamine sulfate group, high-dose, middle-dose, low-dose of Yougui pill treated group (n=10). KOA model was established by modified Hulth method for six weeks. The rats in the high, middle and low-dose of Yougui pill group were treated with Yougui pills at the doses of 20,10 and 5 g/kg respectively by gastrogavage once a day for 8 weeks, while equal volume of normal saline was given to those in the sham and model control group and an equal volume of glucosamine sulfate (1.7 g/kg·d) was given to those in glucosamine sulfate group for 8 weeks. The knee joint was removed after the last dose of drug. The pathological changes of cartilaginous tissues were observed under a microscope. The mRNA levels of Dickkopf homolog 1(DKK1), Wnt induced secreted protein 1(WISP1), Wnt1, low density lipoprotein receptor related protein 5(LRP5) and beta -catenin in rats cartilaginous tissues were analyzed by using RT-PCR method, and the protein contents of DKK1, WISP1, Wnt1, LRP5 and beta-catenin in cartilaginous tissues were detected by Western blot. Results:Compared with the sham group, the articular cartilage was severely damaged, the Mankin score was increased significantly (P<0. 05), the mRNA and protein expression levels of DKK1 in cartilaginous tissue were markedly decreased(P<0.05), while those of WISP, Wnt1, LRP5 and beta-catenin were increased significantly in model group(P<0.05). Compared with model group, the articular cartilage lesions was light (P<0.05), the Mankin Score was decreased significantly(P<0.05), and the mRNA and protein levels of DKK1 in cartilaginous tissue were increased(P<0.05), while those of WISP, Wnt1, LRP5 and beta-catenin were decreased in Yougui pill high-dose group and glucosamine sulfate group (P<0.05). Conclusion:Yougui pill has protective effects on the KOA by inhibiting the expressions of WISP, Wnt1, LRP5, beta-catenin and increasing the expression of DKK1 cytokine in the Wnt signaling pathway.
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    Establishment and application effect of fall prevention and control information system in elderly community
    ZHAO Ting, PI Hong-ying, KU Hong-an, PAN Li, Gong Zhu-yun
    CJAP. 2018, 34 (2): 182-186.   DOI: 10.12047/j.cjap.5540.2018.044
    Abstract   PDF (1133KB) ( 147 )
    Objective:To investigate establishing, applying and evaluating the fall prevention and control information system in elderly community. Methods:Relying on internet technology and informatization means, the fall comprehensive prevention and control strategy of elderly was guided into online from offline. The fall prevention and control information system which was a collection of risk assessment, remote education and feedback was established. One hundred and twenty-six elderly (over 60 years old) in community were screened in this study and 84 high-risk elders who were involved in the remote continuous comprehensive intervention were screened out. Intervening measures included distributing propaganda album, making mission slides and video used to play with the interpretation remotely. Then fall related situation before and after intervention was analyzed and the effectiveness of system evaluated. Results:After remote intervention, the fall incidence of high-risk group decreased from 21.43% to 4.76%(P<0.01). The body balance and gait stability improved clearly(P<0. 01). The rate of taking proper prevention and control behavior significantly improved(P<0.01). They believed in themselves not to fall down with more confidence when taking complex behaviors(P<0.01). The security of environment at home significantly enhanced(P<0. 01). Conclusion:Fall prevention and control information system in elderly community was innovative and convenient. The system could roundly assess the status related to fall and accurately screen out high-risk group. The system could implement the remote continuous comprehensive intervention so that the incident of fall was decrease. In conclusion, the system is stable and effective, can be further popularization and application as a successful pilot.
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    High sensitive ELISA for detection of atrazion
    LI Xiao-li, WANG Ting-ting, LIU Kai, NING Bao-an, MA Xin-hua, LIU Nan, OU Guo-rong, GAO Zhi-xian, LIU Zhong-wen
    CJAP. 2018, 34 (2): 187-192.   DOI: 10.12047/j.cjap.5582.2018.045
    Abstract   PDF (1507KB) ( 102 )
    Objective:To set up ELISA for detection of atrazine with high precision. Methods:The reaction condition of indirect-ELISA was optimized, including atrazine-ovalbumin(AT-OVA) concentration and primary antibody concentration, organic solvent, goat anti-rat immunoglobin G-horseradish peroxidase(IgG-HRP) concentration. The actual samples were detected by the ELISA method established in our laboratory. Then the ELISA method was compared with the HPLC. Results:The specification curve of indirect-ELISA was set up after optimization. The relation coefficient R2=0.9958. The limit of detection (LOD) was 1.972 ng/ml. The percent recovery of the actual samples was in range of 80%~120%. The ELISA detection sensitivity was higher than the HPLC in the range of 0 ng/ml~6 ng/ml atrazine. Conclusion:The ELISA to detect atrazine has good specificity and high precision. And it can be applied in testing real atrazine samples replacing of the large-scale instrument.
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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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