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  28 July 2021, Volume 37 Issue 4 Previous Issue   
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ORIGINAL ARTICLES
Effects of Sox11 gene on neuronal migration in the development mouse cerebral cortex
LYU Ye, SUN Xiao-wan, ZHANG Cong, LUAN Zhi-lin
CJAP. 2021, 37 (4): 337-342.   DOI: 10.12047/j.cjap.6144.2021.052
Abstract   PDF (1788KB)
Objective: To study the effect of the schizophrenia susceptible gene Sox11 on the migration of cortical neurons using mice as experimental animals. Methods: The real-time quantitative PCR and in situ hybridization were used to clarify the expression pattern of Sox11 in the cerebral cortex during development (E14.5, P0, P7, P14). The techniques of plasmid construction, transfection, in utero electroporation and immunostaining were used to explore the role of Sox11 in the neuronal radial migration by transfecting control shRNA plasmid, mSox11 shRNA plasmid and mSox11 shRNA post-interference recovery plasmid in mice of different ages (E17.5, P0, P4, P7). Results: Compared with control neurons, the migration of mSox11 shRNA transfected neurons was delayed significantly. When a part of the neurons in the control group had reached the surface of the neocortex, most of the neurons transfected with mSox11 shRNA remained in the middle area of the neocortex. After the rat Sox11 (rSox11) gene overexpression vector was used to recuse the mouse Sox11 (mSox11) gene interference in mice, the distribution of neurons after migration was basically the same as the control. The distribution of migrating neurons in the subventricular zone (SVZ), intermediate zone (IZ), and cortical plate (CP) was different significantly (P<0.01) after Sox11 interference and recovery. Conclusion: Sox11 can promote the migration of cortical neurons, suggesting that Sox11 plays a crucial role in the migration process of mouse cortical neurons.
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Changes of lncRNA-mRNA co-expression network induced by mtDNA3010A/G genotype mutation in hypoxia
CHEN Yu, GONG Liang, LUO Yong-jun
CJAP. 2021, 37 (4): 343-348.   DOI: 10.12047/j.cjap.6076.2021.038
Abstract   PDF (1401KB)
Objective: To analyze the long non-coding RNA (lncRNA) and messenger RNA (mRNA) co-expression network changes induced by mtDNA3010A/G mutation in acute hypoxia, and to investigate the role of key lncRNA and mRNA in the regulation of gene expression induced by hypoxia. Methods: The genotype combinations A-C-C and G-C-C of mitochondrial DNA 3010-5178-10400 were screened, and genotypes of mtDNA3010A and mtDNA3010G fusion cells were constructed by using osteosarcoma cell treated by ethidium bromide without mitochondrion (ρ0206 cell) as donors. After treatment with 1% O2 24 h, the lncRNA - mRNA expression chip was applied to detect the differently expressed lncRNA and mRNA in two kinds of fusing cells, and fluorescence quantitative polymerase chain method was used to verify differently expressed mRNA. Bioinformatics methods were applied to build co-expression network of lncRNA-mRNA, predict target genes of differently expressed lncRNA, and the functions of differently expressed mRNA and target genes predicted by lncRNA were also analyzed based on gene ontology (GO) and the Kyoto encyclopedia of genes and genomes (KEGG) forecast analysis. Results: After treatment with 1% O2 for 24 h, compared with mtDNA3010G fusion cells: 688 lncRNAs were up-regulated, 21 were more than 2 times; 1098 were down-regulated, and 4 were more than 2 times. There were 1151 mRNA expressions up-regulated, 14 were more than 2 times, 539 mRNA expressions were down-regulated, and 3 were more than 2 times. Conclusion: MtDNA3010A/G genotype mutation under hypoxia is able to affect the lncRNA-mRNA regulatory network, and the differentially expressed lncRNA and mRNA may play an important role in regulation network of gene expression induced by hypoxia, which is expected to be a target for the regulation of hypoxia reaction from the perspective of mitochondria.
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Effects of Notch3 on gene expression and signal pathway of pancreatic stellate cell activation
SONG Hai-yan, ZHOU Zhi-xin, ZHANG Yu-xiang
CJAP. 2021, 37 (4): 349-353.   DOI: 10.12047/j.cjap.6084.2021.024
Abstract   PDF (1124KB)
Objective: The pancreatic stellate cells( PSCs) of mice were isolated and cultured and the effects of Notch3 siRNA on PSCs gene expression were detected. Methods: were PSCs of mice were isolated and cultured. The expressions of α-SMA, fibonectin and collagen I in activated PSCs were detected by immunofluorescence. The PSCs were divided into four groups, blank control group (MOCK group), negative siRNA control group (NC group) , Notch3 siRNA group (N3 siRNA group) and Notch3 siRNA-1 group (N3 siRNA-1 group). Cell treatment: the same transfection method was applied to transfect PSCs for 48h. Then total RNA was extracted from each group, and the concentration and purity of RNA was measured. The transcriptome sequencing and analysis were performed by ANOROAD Gene technology (Beijing) co., LTD. Results: The immunofluorescence results showed that α-SMA, fibonectin and collagen I were significantly expressed in activated PSCs. The analysis of the sequencing results showed that the gene expressions of α-SMA, collagen I, fibronectin, CTGF and PCNA of PSCs were down-regulated in N3 siRNA group and N3 siRNA-1 group comparing with NC group. The genes involved in collagen metabolism were up-regulated, the gene expression of positive regulation of collagen biosynthesis was down-regulated, while that of negative regulation of collagen biosynthesis was up-regulated in N3 siRNA group and N3 siRNA-1 group comparing with NC group. The genes that regulated cell aggregation were down-regulated, and the genes that regulated extracellular matrix were down regulated in N3 siRNA group and N3 siRNA-1 group comparing with NC group. Inhibition of Notch3 expression in PSCs could affect the gene expressions of cell adhesion molecule signaling pathway, MAPK signaling pathway and TGF-β signaling pathway. Conclusion: Inhibition of Notch3 expression can inhibit activation of PSCs , and reduce the ability of proliferation, migration and aggregation and ECM synthesis of PSCs. Inhibition of Notch3 expression may affect other signal pathways such as cell adhesion molecule signaling pathway, MAPK signaling pathway and TGF-β signaling pathway, but its effects need further validation.
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Effects of GPR81 agonist on insulin resistance in rats with nonalcoholic fatty liver disease
ZHANG Yu, LU Qing-hua, CAO Hai-fang, ZHANG Sheng-rong, WANG Hu-de
CJAP. 2021, 37 (4): 354-358.   DOI: 10.12047/j.cjap.6055.2021.033
Abstract   PDF (1971KB)
Objective: To investigate the effects of NOD-like receptor protein 3 (NLRP3) signaling pathway on insulin resistance and the intervention of lactic acid receptor G protein-coupled receptor 81 (GPR81) agonist in nonalcoholic fatty liver disease (NAFLD) rats. Methods: Thirty SD male rats were randomly divided into three groups: control group, NAFLD group and GPR81 agonist group, with 10 rats in each group. Nonalcoholic fatty liver rat model was established by high fat diet. The rats in GPR81 agonist group were injected intraperitoneally with GPR81 specific agonist lactate (50 nmol/L) on the basis of nonalcoholic fatty liver model once a week, and the other two groups were injected with the same amount of normal saline for 12 weeks. The levels of liver biochemical indexes, fasting blood glucose, insulin and inflammatory factors in liver homogenate were measured, and the histopathological morphology of liver in each group was observed. The protein expressions of NLRP3, apoptosis-associated speck-like protein containing a CARD (ASC), cysteinyl aspartate specific proteinase-1 (caspase-1), insulin receptor substrate 1 (IRS-1), Tyr465-IRS-1, Ser636-IRS-1, glucose transporter 4 (GLUT4) in liver tissue were detected by Western blot. The mRNA expression levels of NLRP3, ASC, caspase-1, IRS-1 and GLUT4 in liver tissue were detected by qRT-PCR. Results: Compared with the control group, the serum levels of triglyceride (TG), alanine aminotransfease (ALT), aspartate aminotransfease (AST), fasting plasma glucose (FPG), fasting insulin (FINS) and homeostasis model assessment of insulin resistance (HOMA-IR) of NAFLD group were increased significantly (P< 0.05). The results of liver histopathology showed that in NAFLD group, there were obvious fatty changes in liver tissue, fat droplets in hepatocytes and inflammatory cell infiltration. And the mRNA and protein expressions of NLRP3, ASC, caspase-1 and the protein expression of Ser636-IRS-1 in NAFLD group were increased significantly, and the contents of interleukin 1β (IL-1β) and interleukin 18 (IL-18) in liver and serum were increased, while the mRNA and protein expressions of IRS-1 and GLUT4 and Tyr465-IRS-1 were decreased significantly (P<0.05). Compared with NAFLD group, the above indexes of GPR81 agonist group were all significantly improved. Conclusion: The activation of NLRP3 signaling pathway mediates the production of inflammatory factors and promotes the development of NAFLD. GPR81 agonist may be a potential treatment for NAFLD.
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Effects of acupuncture on endoplasmic reticulum pathway in exercise-induced skeletal muscle damage in rats and its mechanism
DING Hai-li, HUANG Zeng-hao, REN Zai-fang, SUN Zhu-xin, LI Jun-ping, WANG Rui-yuan
CJAP. 2021, 37 (4): 359-364.   DOI: 10.12047/j.cjap.6097.2021.044
Abstract   PDF (2005KB)
Objective: To observe the effects of acupuncture on the endoplasmic reticulum (ER) functional enzymes sarcoplasmic reticulum Ca2+-ATPase (SERCA), protein disulfide isomerase (PDI), glucose-regulated protein 78(GRP78) and PERK pathways in rats with exercise-induced skeletal muscle damage, and to explore the mechanisms of acupuncture in preventing and treating exercise-induced skeletal muscle damage. Methods: Eight-week-old male SD rats were randomly divided into control group (group C, n=6), exercise group (group E, n=30), acupuncture group (group A, n=30) and exercise acupuncture group (group EA, n=30). Among them, the E and EA group were established an exercise-induced skeletal muscle damage model by a single eccentric exercise, and acupuncture intervention was applied 0.5 cm above the Achilles tendon of the rat's calf immediately after EA exercise, and in group A, acupuncture intervention was applied during the same period. Each group was divided into 0 h/12 h/24 h/48 h/72 h (n=6) according to different sampling time points after exercise and acupuncture intervention, and soleus muscle was collected at the corresponding time for index test. The ultrastructure of muscle fibers was observed by transmission electron microscopy; the contents of SERCA and PDI were determined by ELISA; and the expressions of ER stress marker proteins GRP78 and p-PERK and p-eIF2α were detected by Western blot. Results: Compared with group C, there were no significant differences in the indicators of group A at all time points (P> 0.05), the ultrastructure of muscle fibers in group E showed different damages, SERCA content was significantly decreased from 0 h to 48 h (P<0.05), PDI content was significantly increased from 0 h to 72 h (P<0.05), GRP78 expression was significantly increased from 0 h to 72 h (P<0.05), p-PERK expression was significantly increased from 0 h to 24 h (P<0.05), and p-eIF2α expression was consistent with p-PERK. Compared with the corresponding times in group E, the ultrastructure of muscle fibers in group EA was significantly alleviated, SERCA content was significantly increased from 48 h and 72 h (P<0.05), PDI content was significantly increased from 0 h to 72 h (P<0.05), and GRP78 expression was significantly decreased from 0 h to 72 h (P<0.05). Conclusion: Acupuncture can effectively ameliorate exercise-induced skeletal muscle damage and alleviate ER stress after a large load eccentric exercise. The mechanism of them may be related to the up-regulation of protein disulfide isomerase PDI and the inhibition of ER stress PERK pathway.
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Effects of electroacupuncture on autophagy factors and liver lipid metabolism in rapidly aging mice
YANG Yun- hao, PANG Fang, HUANG Si-qin, YANG Zhi-xue, ZHU Zheng-wei, DAI Pan, GUO Xiao, LIAO Dong-mei, TANG Cheng-lin
CJAP. 2021, 37 (4): 365-370.   DOI: 10.12047/j.cjap.6156.2021.053
Abstract   PDF (1939KB)
Objective: To study the effects of electroacupuncture on the expressions of autophagy-related factors LC3-Ⅱ, Beclin1, Atg7, and P62 in the liver of rapidly aging (senescence accelerated mouse/prone8,SAMP8) mice, and to explore the mechanisms of electroacupuncture to improve liver lipid metabolism in mice. Methods: Thirty-week-old male SAMP8 mice were randomly divided into model group, drug group, and electroacupuncture group, with 7 mice in each group. Seven anti-rapid aging SAMR1 mice of the same age were used as the control group. The animals in the control group and the model group were bred routinely for 2 weeks without any intervention; the drug group was treated with intraperitoneal injection of rapamycin at the dose of 10 mg·kg-1·d-1, once a day, 6 consecutive days a week; the electroacupuncture group was given "Shenshu" and "Taichong" Electroacupuncture at point(15 minutes a day, 6 consecutive days a week). The serum lipid metabolism and liver lipid deposition of mice were detected, the distribution of liver autophagy body, the protein and mRNA expressions of liver LC3 - Ⅱ, Beclin1, Atg7 and P62 were determined. Results: Compared with the control group, the total cholesterol (TC), triglycerides (TG), and low density lipoprotein (LDL) of the model group were increased significantly(P<0.01). In the model group, lipid droplet deposition was obvious, autophagosomes were decreased, the protein and mRNA expression levels of autophagy- related factors LC3-Ⅱ, Beclin1 and Atg7 were decreased significantly (P<0.01), while the protein and mRNA expressions of P62 were increased significantly (P<0.01). Compared with the model group, the serum contents of TG, TC, and LDL of the mice in the electroacupuncture group and the drug group were decreased significantly (P<0.01), lipid droplet deposition was reduced, autophagosomes were increased, the protein and mRNA expression levels of LC3 -Ⅱ, Beclin1 and Atg7 were increased significantly(P<0.01), and the protein and mRNA expression levels of P62 were decreased significantly(P<0.01). The protein and mRNA expression levels of Beclin1 and Atg7 in the liver of the electroacupuncture group were not significantly different from the drug group (P>0.05). Conclusion: Electroacupuncture can alleviate liver lipid metabolism disorders, which may be related to the regulation of the expressions of liver autophagy related factors LC3-Ⅱ, Beclin1, Atg7, and P62, thereby promoting liver autophagy in SAMP8 mice.
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Characteristics of circulation and microcirculation in healthy people of Han nationality at different altitudes
HE Zong-zhao, DENG Li, MA Si-qing, LI Xin-hui, WANG Hao
CJAP. 2021, 37 (4): 371-375.   DOI: 10.12047/j.cjap.6014.2021.006
Abstract   PDF (1001KB)
Objective: To observe the circulation and microcirculation characteristics of healthy han people in Qinghai at different Guoluo (4 100 m) and Huzhu (2 600 m) and in Shanghai in plain area (4 m). Methods: The 28 healthy han people aged (45.62±10.15) from Guoluo in Qinghai, 27 healthy han people aged (47.25±8.43) from Huzhu in Qinghai and 20 healthy han people aged (43.12±8.28) from Shanghai were divided into three groups: Guoluo group (GL), Huzhu group (HZ) and Shanghai group (SH); Venous blood was collected for routine blood test: red blood cells count(RBC), hemoglobin(HGB), hematocrit(HCT), platelet(PLT); and coagulation index: prothrombin time(PT), interntionl normlized ratio(INR), fibrin(Fib), activated partial thromboplastin time (APTT), thrombin time(TT), continuous non - invasive hemodynamic monitor (CNAP) was used for continuous measurement: cardiac output (CO), heart rate (HR), stroke volume (SV), pulse pressure variation rate (PPV), systemic vascular resistance index (SVRI), mean arterial pressure (MAP); pulse oxygen monitoring instrument is used for monitoring SPO2 (pulse oxygen); the total vascular density (TVD), perfusion vascular density (PVD), proportion of perfused vascular(PPV), and microvascular flow index (MFI) of sublingual microcirculation were observed by using peripheral dark field imaging (SDF) monitoring technique. Results: Compared with the Shanghai group, the RBC and HGB levels in the Huzhu group and the Guoluo group were significantly increased (P<0.05), while PLT levels were significantly decreased (P<0.05); The levels of PT, Fib, APTT, TT, CO, HR, SV, MAP and SPO2 in the Huzhu group were significantly decreased (P<0.05), while the levels of TVD, PVD, PPV and MFI were significantly increased (P<0.05). In Guoluo group, the levels of HCT, Fib, CO, SV, PPV, MAP, TVD and PVD were significantly increased (P<0.05), while the levels of PT, INR, TT and SPO2 were significantly decreased (P<0.05). Compared with the Huzhu group, the levels of RBC, HGB, HCT, Fib and APTT of Guoluo group were significantly increased (P<0.05); while the levels of SPO2, TVD, PVD, INR and TT were significantly decreased (P<0.05). Conclusion: The microcirculation of healthy people in different altitude areas is significantly different, and the microcirculation density of healthy people in high altitude areas is significantly higher than that in plain areas, especially in moderate altitude areas. Its special physiological significance is of guiding significance for the treatment of infectious shock and chronic altitude disease in plateau areas.
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Effects of Mongolian medicine Shaosha-7 on myocardial ischemia/ reperfusion injury of rats
YANG Xiang-jun, WANG Yu, E.bayarima, ZHAO Ming, S. hexigezarigala
CJAP. 2021, 37 (4): 380-384.   DOI: 10.12047/j.cjap.6114.2021.029
Abstract   PDF (1712KB)
Objective: To investigate the effects of Shaosha-7 in rats with myocardial ischemia-reperfusion injury and its mechanisms. Methods: male SD rats were divided into sham operation group (n=10), myocardial ischemia-reperfusion injury group (n=10), low, medium and high dose of Shaosha-7 groups (n=10), and positive drug group (n=10). The rats of Shaosha-7 (low, medium and high dose) groups were treated with Shaosha-7 at the doses of 0.4, 0.8 and 1.6 g/kg respectively, once a day for 15 days. The rats of positive drug group were treated with 0.3 g/kg Danshen, once a day for 15 days. The rats of the sham operation group and myocardial ischemia-reperfusion injury were treated with 2 ml/100 g distilled water, once a day for 15 days. After 15 days, the rats of the model group and the treatment group underwent thoracotomy and ligation of coronary artery for 30 minutes, then thoracic cavity was closed after reperfusion. Rats in six groups were executed electrocardiographic examination and their hearts were taken for Hematoxylin-Eosin (HE) staining and 2, 3, 5-triphenyltetrazolium chloride (TTC) staining to observe infarct area and myocardial pathological changes. The contents of cTnI, CK-MB, LDH, MDA, SOD, GSH-Px, TNF-α, IL-18, IL-6 and IL-1 β in serum were detected by ELISA. The expression of NF-кB was detected by immunohistochemistry. Results: Compared with the sham operation group, the infarct size, the levels of cTnI, CK-MB, CK-MB, LDH, MDA, GSH-Px, TNF-α, IL-18, IL-6, IL-1β and NF-кB were increased and the content of SOD were decreased in rats with myocardial ischemia-reperfusion injury. Compared with the rats with myocardial ischemia-reperfusion injury, Shaosha-7 improved the arrhythmia and pathological changes, reduced the infarct area, decreased the contents of cTnI, CK-MB, LDH, MDA, GSH-Px, TNF-α, IL-18, IL-6, IL-1 β, increased the content of SOD, decreased the expression of NF-кB. Conclusion: Mongolian medicine Shaosha-7 can effectively alleviate myocardial ischemia-reperfusion injury in rats. This study provides a theoretical basis for the treatment of myocardial ischemia-reperfusion (I/R) injury with Shaosha-7.
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The regulatory role of autophagy in rats lung ischemia/reperfusion injury
HAO Mao-lin, LOU Guo-qiang, LIU Xiu-jie, QIAN Wei, WANG Jia, ZHOU Zhuo-lin, WANG Wan-tie
CJAP. 2021, 37 (4): 385-388.   DOI: 10.12047/j.cjap.6061.2021.019
Abstract   PDF (1448KB)
Objective: To investigate the role of cell autophagy in lung ischemia/reperfusion injury in rats. Methods: Forty SD rats were randomly divided into 5 groups (n=8): ①Sham operated group (sham group):just open rat chest for 3.5 h; ②Ischemia/reperfusion group (I/R group):after open chest, clamp pulmonary hilus for 0.5h then reperfusion for 3 h; ③Solvent group (DMSO group): intraperitoneal injection of DMSO solution for 1h before operation; ④Autophagic inhibitor group (3-MA group); ⑤Autophagic agonist group (Rap group): intraperitoneal injection of autophagic agonist rapamycin before operation; the rest operations of DMSO, 3-MA and Rap groups are the same as that of I/R group. At the end of the experiment, the rats were killed by euthanasia-killing. The lung tissues were collected and the wet/dry weight ratio (W/D) and total lung water content (TLW) of the lung tissues were detected. The lung tissue structure and cell ultramicro morphology were observed by light microscopy and electron microscopy and the injuried alveolar rate(IAR) was calculated. The autophagy-related protein expressions were detected by Western blot. Results: Compared with sham group, the levels of W/D, TLW and IAR were increased, the expressions of autophagy related protein and p-AMPK, Beclin 1, LC3 II were also increased in other four groups, while the protein expressions of p-mTOR and p62 were decreased significantly (P< 0.05 or P<0.01). Under the light microscope, the other groups of lung tissue had edema and exudation in varying degrees, the structure of alveoli was disordered, the ultrastructural damage of cells was aggravated under the electron microscope, and autophagosome could be observed. Compared with DMSO group, the expressions of autophagy related protein, the levels of W/D, TLW and IAR in 3-MA group were decreased (P<0.05 or P<0.01), the edema of lung interstitial was lighter, and less cells were found in alveolar cavity. Ultrastructural damage was also lighter and with less autophagosome. Besides, there was no significant difference among I/R, DMSO and Rap groups (P>0.05). Conclusion: Autophagy can be activated during ischemia/reperfusion in rats to induce lung injury.
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Changes of FLI-1 protein expression in mice with pulmonary endothelial barrier dysfunction following acute lung injury induced by lipopolysaccharide
XU Li-cong, HUANG Ting-ting, HAO Jia-le, FAN Xiao-fang, GONG Yong-sheng, MAO Sun-zhong
CJAP. 2021, 37 (4): 393-396.   DOI: 10.12047/j.cjap.6080.2021.040
Abstract   PDF (1083KB)
Objective: To observe changes of Friend leukemia virus integration 1 (FLI-1) protein expression of pulmonary tissue in mice with pulmonary endothelial barrier dysfunction following acute lung injury (ALI) induced by lipopolysaccharide (LPS). Methods: The mouse model of ALI was established by injection of LPS (7.5 mg/kg, i.p. ). At 0 h, 12 h, 24 h and 48 h after LPS injection, pulmonary microvascular endothelial permeability and lung wet/dry weight ratio (W/D) were assessed. The contents of TNF-α and IL-6 in bronchoalveolar lavage fluid (BALF) were detected by ELISA method. The protein levels of FLI-1 and Src protein tyrosine kinase (SRC) were analyzed by Western blotting.Results: ①Pulmonary microvascular endothelial permeability at 12 h and 24 h were significantly higher than those of 0 h by 74.3% and 162.4%, respectively, while that of 48 h was lower than that of 24 h by 27.0% (P<0.05). The W/D at 12 h and 24 h were significantly higher than those of 0 h by 50.1% and 122.9%, respectively, while that of 48 h was lower than that of 24 h by 10.7% (P<0.05). ②The contents of IL-6 and TNF-α in BALF at 12 h and 24 h were significantly higher than those of 0 h, while those of 48 h were significantly lower than those of 24 h by 28.3% and 21.6% (P<0.05), respectively. ③The protein level of FLI-1 in lung at 12 h and 24 h were down-regulated than those of 0 h by 20.4% and 56.9%, respectively, while that of 48 h was up-regulated than that of 24 h by 18.2% (P<0.05). The protein level of SRC in lung at 12 h and 24 h were up-regulated than those of 0 h by 76.8% and 176.7%, respectively, while that of 48 h was down-regulated than that of 24 h by 33.4% (P<0.05).④Same as the protein level of FLI-1 with the protein level of SRC in lung, pulmonary microvascular endothelial permeability was significantly negative correlated with the protein level of FLI-1 in lung, while it was significantly positive correlated with the protein level of SRC (P<0.01). Conclusion: FLI-1 participates in the pathological proceeding of pulmonary endothelial barrier dysfunction following ALI induced by LPS.
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Protective effects of total saponins of Codonopsis on ulcerative colitis induced by TNBS in rats and its mechanism
LIU Xue-feng, QIAO Jing, GAO Jian-de, CHEN Zheng-jun, LIU Xiong
CJAP. 2021, 37 (4): 397-401.   DOI: 10.12047/j.cjap.6051.2021.032
Abstract   PDF (1807KB)
Objective: To study the protective effects and mechanisms of total saponins of Codonopsis (TSC) on ulcerative colitis in rats. Methods: Fifty male Wistar rats were randomly divided into 5 groups: control group, model group, salazosulfadiazine (SASP) positive control group (0.3 g/kg), TSC high- and low-dose experimental groups(1.2, 0.4 g/kg). UC rat model was established by trinitrobenzene sulfonic acid (TNBS)/ ethanol enema. After administration for 21 days, the rats' symptoms and signs, disease activity index (DAI), colonic mucosal injury index (CMDI) and colonic tissue morphology were observed. The contents of superoxide dismutase (SOD), malondialdehyde (MDA), inflammatory cytokines interleukin-6 (IL-6), interleukin-10 (IL-10) and tumor necrosis factor (TNF-α) in colon tissues were determined. Protein expression of nuclear nuclear transcription factor-κB (NF-κB) in colon tissues was detected. Finally, the effect of TCS therapy was evaluated. Results: Compared with the control group, the DAI and CMDI scores of the rats in the model group were increased significantly, meanwhile the colonic mucosa was seriously damaged, indicating that the model was successful. Compared with the model group, the TSC high and low dose groups could significantly reduce the DAI and CMDI score (P<0.05) and improve the colonic mucosa form. TSC also could increase the SOD activity and decrease MDA content in colon tissues(P<0.05), while inhibit the levels of IL-6 and TNF-α mRNA in the colon tissues and promote the expression of IL-10 mRNA (P<0.01). At the same time, TSC reduced the expressions of NF-κB protein in the colon (P<0.01). The TSC high-dose group was superior to the low-dose group (P<0.05). Conclusion: TSC has significant protective effects on ulcerative colonic mucosal damage in UC rats, and there is a dose-dependent relationship; its mechanism may be related to anti-lipid peroxidation and inhibiting the NF-κB signaling pathway to regulate the release of inflammatory factors.
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Effects of miR-335-5p targeting G6PD on proliferation and apoptosis of colon cancer cells
FAN Xiao-qun, LI Huan, ZHU Hua-xiong, HUANG Jian-ping, HE Lyu-fen
CJAP. 2021, 37 (4): 402-406.   DOI: 10.12047/j.cjap.6090.2021.041
Abstract   PDF (1097KB)
Objective: To investigate the effects of miR-335-5p targeting glucose-6-phosphate dehydrogenase (G6PD) on the proliferation and apoptosis of colon cancer cells. Methods: Normal colon cell group, blank control group, NC group and miRNA-335-5p mimic group were set up. Colonic epithelial cells (IEC) and human colon cancer cells SW480 were cultured in vitro, and the cells in the NC group and miRNA-335-5p mimic group cells were transfected. RT-qPCR was used to detect the expression levels of miR-335-5p and G6PD mRNA in each group of cells. The targeting effect of miR-335-5p on G6PD was verified by Double Luciferase Report experiment. MTT assay was used to detect cell proliferation. Flow cytometry was used to detect the apoptosis rate. The expressions of G6PD, Bax, Bcl-2 and caspase-3 were detected by Western blot. Results: Compared with normal colon cells, the relative expression levels of miR-335-5p in SW480 cells of colon cancer in the blank control group and NC group were decreased, and the relative expression level of G6PD mRNA was increased (P<0.05); compared with the blank control group and NC group, the expression level of miR-335-5p in miR-335-5p mimic group was increased significantly, and the expression of G6PD mRNA was decreased significantly (P<0.05). Compared with the blank control group and NC group, the proliferative activity of colon cancer SW480 cells in miR-335-5p mimic group was decreased significantly, and the apoptosis rate was increased significantly (P<0.05). The relative activity of luciferase in miR-335-5p mimic + WT-G6PD 3 '- UTR group was lower than that in miR-335-5p NC + WT-G6PD 3' - UTR group (P<0.05). Compared with the blank control group, the relative expression levels of G6PD and bcl-2 protein in miR-335-5p mimic group were decreased significantly, and the expression levels of Bax and caspase-3 protein were increased significantly (P<0.05). Conclusion: MiR-335-5p may inhibit the proliferation and promote apoptosis of colon cancer cells by targeting G6PD.
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Clinical and prognostic features of the patients with esophageal squamous cell carcinomas as a first primary malignancy
WEN Zhen, ZHANG Yan-qiu, WU Rong, PENG Xue-mei, CHEN Bin, LENG Ai-min
CJAP. 2021, 37 (4): 407-414.   DOI: 10.12047/j.cjap.6020.2021.008
Abstract   PDF (1409KB)
Objective: To retrospectively analyze the clinical characters and prognosis of the patients with Esophageal Squamous Cell Carcinomas as First Primary Malignancy (ESCCFPM), which will help us better understand the relationship between Esophageal Squamous Cell Carcinoma (ESCC) and other cancers, and to provide appropriate research evidence for the clinical diagnosis and treatment. Methods: The clinicopathological and follow-up data of 540 Patients with ESCCFPM between January 1, 2004 and December 31, 2016 were collected from the Surveillance, Epidemiology and End Results (SEER) database of National Cancer Institute. The Kaplan-Meier method was used to determine Overall Survival (OS) curves of ESCC patients, and the Log-Rank test was used to estimate differences in survival. The Cox proportional hazards models were adopted for the prognosis analyses. Results: Regarding the number of multiple primary malignancies (MPMs), 491 had two malignancies, 42 had three malignancies and 7 had four malignancies. ESCCFPM is more common among males. The high incidence age is between 61 and 80 years old. Tumors of the respiratory system (36.9%), were the most common MPMs followed by digestive system (35.2%) and reproductive system (8.9%). The 1-year, 3-year, 5-year OS rates for patients with ESCCFPM were 76.9%, 50.4% and 38.9%, respectively. The age of the ESCC diagnosed, T stage, time of occurrence, carcinoma number, lymph node dissection, surgery, radiotherapy and chemotherapy were the prognostic factor of overall survival for ESCCFPM patients. Age, race, T stage, time of occurrence surgery and radiotherapy were independent prognostic factors for the whole cohort by multivariate survival analysis. Conclusion: ESCCFPM,mainly two-lesion cancer, is most commonly found in respiratory system and digestive systems. Enhanced follow-up of respiratory and digestive tumors in ESCCFPM patients aged 61-80 may help identify multiple primary malignancies. Surgery, radiotherapy and chemotherapy may improve overall survival for ESCCFPM patients.
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The effects of shikonin on liver cancer cells SMMC-7721 apoptosis and its mechanism
WANG Yu, HE Shi-yan, ZHU Ruo-ting, KE Rui-jun, CHEN Jia-yu
CJAP. 2021, 37 (4): 415-418.   DOI: 10.12047/j.cjap.6137.2021.051
Abstract   PDF (1492KB)
Objective: To investigate the effects and molecular mechanisms of shikonin on liver cancer SMMC-772 cells. Methods: SMMC-7721 cells were treated with shikonin at the concentrations of 0, 5, 20, 80 and 320 ng/ml for 0, 24, 48 and 72 h respectively. The proliferative activity of the cells was detected by CCK8 assay. The nuclear type changes of cells was observed after hoechst 33342 staining. Flow cytometry was used to analyze cell apoptosis and death rate. The expressions of proteins in cells were determined by Western blot, and the tumor inhibitive effects were observed through anti-tumor experiment on the BALB/c mice. Results: In vitro experiments, shikonin could inhibit the proliferation of SMMC-7721 cells and induce their apoptosis(P<0.01), up-regulate the expression of p53 gene, down-regulate the phosphorylation levels of AKT and PI3K protein. In vivo study also confirmed that shikonin could significantly inhibit the growth of tumor in tumor-bearing mice(P<0.01)in dose-dependent and time-dependent manners. Conclusion: Shikonin can inhibit the proliferation activitity and induce apoptosis of SMMC-7721 cells by affecting the PI3K/AKT signal pathway and has potential anti-liver cancer functions.
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Effects of betulinic acid on apoptosis of human gastric cancer MGC-803 cells
FAN Hua, SHAO Shu-li, HE Meng-qi, HUANG Xin , ZHANG Wei-wei, ZHANG Zhen-zhu
CJAP. 2021, 37 (4): 419-422.   DOI: 10.12047/j.cjap.6078.2021.039
Abstract   PDF (2034KB)
Objective: The effects of betulinic acid (BA) on apoptosis of human gastric cancer MGC-803 cells was investigated by using human gastric cancer MGC-803 cells as experimental materials, and the basis for its clinical application was provided. Methods: The human gastric cancer MGC-803 cells were divided into 4 groups,each group was set with 3 replicates.The control group was MGC-803 cells without being added betulinic acid; the other 3 groups of experimental groups were treated with betulinic acid at final concentrations of 10, 20 and 30 μg /ml respectively. Cells were treated with betulinic acid of different concentrations for 48 h. Laser confocal microscope was used to observe morphological changes of MGC-803. The activities of Caspase-3 and Caspase-9 were detected by an assay kit. Flow cytometry was applied to determine mitochondrial membrane potential. The mRNA and protein levels of Caspase-3, Caspase-9 and Cyt c were also detected by qRT-PCR and Western blot, respectively. Results: Compared with the control group, the activities of Caspase-3 and caspase-9 were increased(P<0.01), while the mitochondrial membrane potential was decreased significantly(P<0.01). The mRNA and protein expressions of Caspase-3, caspase-9 and Cyt c were up-regulated significantly(P<0.01). Conclusion: In the final concentration range of 10 ~ 30 μg/ml, betulinic acid can induce apoptosis of human gastric cancer MGC-803 cells by regulating the expression of Caspase-3, Caspase-9 and Cyt c.
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Progranulin attenuated asthma inflammation by inhibiting the expression of IL-6
QI Yu-hong, SUN Si, WANG Ling, YANG Jing, WU Guang-ying, HUANG Qi, XU Wen-chun
CJAP. 2021, 37 (4): 423-428.   DOI: 10.12047/j.cjap.6126.2021.050
Abstract   PDF (1551KB)
Objective: To investigate the role and mechanism of progranulin (PGRN) in asthma. Methods: Control group and model group were set up in wild and IL-6 knockout (IL-6 ko) mice, respectively. For asthma model, mice were intraperitoneally sensitized with 100 μg OVA on days 0 and 7, followed by aerosol challenges with 5% OVA for 30 min per day from day 14 to 21, and mice were sacrificed 24 h after the last challenge. The mice in control group were treated in the same way with PBS. Bronchoalveolar lavage fluid (BALF) was collected for leukocytes count and differential count. The pathological changes of lung tissues were observed by H&E staining. The cytokines in lung homogenate, serum and BALF were detected by Q-PCR and ELISA. The in vitro model of asthma was induced by stimulating A549 or BEAS-2B cells with IL-13. Each group was replicated in three wells and four groups were designed: PBS group, IL-13 treatment group, IL-13 + rhPGRN treatment group, inhibitors of p38 phosphorylation (SB203508) treatment group. The cells or supernatant were collected after 0~48 h. PGRN and IL-6 levels were determined by Q-PCR and ELISA, the level of p38 phosphorylation was tested by Western blot (WB). Results: Compared with control group, PGRN levels were decreased in lung homogenate and BALF (P<0.05), and PGRN presented a downtrend in serum, however, the level of IL-6 in BALF was increased in asthma mice (P<0.01). In IL-6 ko asthma mice, compared with the wild asthma mice, leukocytes, especially neutrophils in BALF were decreased (P<0.05), but PGRN was increased (P<0.05), lung pathological damage was significantly alleviated. In vitro experiments, compared with PBS group, PGRN level was decreased (P<0.05), IL-6 level was increased (P<0.01), phosphorylation of p38 was activated in IL-13 treatment group. Compared with IL-13 treatment group, in IL-13 + PGRN treatment group, IL-6 level was decreased (P<0.05); phosphorylation of p38 was inhibited (P<0.05); and the production of IL-6 (P<0.05) was decreased after treatment with inhibitor of p38 phosphorylation. Conclusion: PGRN inhibited the production of IL-6 by suppressing the p38 phosphorylation to alleviate asthmatic airway inflammation.
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Effects of astaxanthin combined with aerobic exercise on renal aging of rat induced by D-galactose and its mechanism
NIU Yan-long, CAO Jian-min, ZHOU Qi-yun, HU Ge, GUO Xian, LIU Jian, HAO Min, ZHANG Tao, ZHOU Hai-tao
CJAP. 2021, 37 (4): 433-438.   DOI: 10.12047/j.cjap.6064.2021.034
Abstract   PDF (2000KB)
Objective: To study the effects and mechanisms of astaxanthin combined with aerobic exercise on renal senescence of rat induced by D-galactose. Methods: Sixty 3-month-old SPF SD rats were divided into control group (C group), acute senescence group (S group), astaxanthin+acute senescence group (AS group), aerobic exercise+acute senescence group (ES group), astaxanthin+aerobic exercise+acute senescence group (AES group), by two-factor two-level 2×2 factorial design with 12 rats in each group. Acute senescence model of rat was establshed by intraperitoneal injection with 100 mg/(kg·d) D-galactose, and the intervention was conducted with 20 mg/(kg·d) astaxanthin and/or aerobic exercise with 60% VO2max for 6 weeks. The histopathological/ultrastructural changes of the kidney were observed by light microscope/electron microscope; the levels of SOD, γ-GCS and MDA were detected by ELISA, and LDF in kidney was determined by fluorescence colorimetry; the protein expression of Nrf2 signaling pathway was detected by immunohistochemistry. Results: Compared with AS and ES group, in AES group, the improvement of renal tissue morphology/ultrastructure was more significant; LDF was decreased significantly (P<0.01); SOD activity was significantly increased (P<0.01); γ-GCS was significantly higher than that of AS group, but not significantly different from that of ES group (P>0.05); there was no significant difference in MDA between groups (P>0.05); the levels of Nrf2 and p-Nrf2 were increased significantly (P<0.05, P<0.01); HO-1 was significantly higher than that of ES group(P<0.05), but not significantly different compared with that of AS group(P>0.05). Conclusion: Astaxanthin combined with aerobic exercise can delay aging process of kidney, its mechanism may be that the combination regulate the protein expression in Nrf2 signaling pathway, Ⅱ detoxifying enzymes and antioxidant enzyme activity, and improve oxidative stress in kidney of rat induced by D-galactose.
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Effects of aerobic interval training on myocardial oxidative stress and inflammation in rats with myocardial infarction and its mechanism
GENG Yuan-wen, LIN Qin-qin, WANG Xiang-yi, LI Ruo-ming, TIAN Zhen-jun
CJAP. 2021, 37 (4): 439-444.   DOI: 10.12047/j.cjap.6094.2021.043
Abstract   PDF (1296KB)
Objective: The present study was to determine the effects of aerobic interval training (AIT) on the expressions of SIRT1, Nox4 and inflammatory factor in the heart of rats with myocardial infarction (MI). Methods: Male Sprague Dawley rats were randomly divided into sham-operated group (C), sedentary MI group (MI) and MI with AIT group (ME) (n=10). The MI model was established by ligation the left anterior descending coronary artery. Rats in C groups were subjected to the same surgery, but only threaded and not ligated. After surgery 1 week, rats in ME groups took adaptability training for 1 week, and then subjected to 4 weeks treadmill exercise training. After training, the hearts were collected for histological observation. The level of malondialdehyde (MDA) and the activity of superoxide dismutase (SOD) in heart was assessed by ultraviolet spectrophotometry. The activity of lactate dehydrogenase (LDH) was determined by enzyme linked immunosorbent assay. The expression of sirtuin1 (SIRT1) mRNA was examined by real-time quantitative polymerase chain reaction (RT-qPCR). The protein expressions of SIRT1, NADPH oxidase 4 (Nox4), tumor necrosis factor-alpha (TNF-α) and interleukin-1β (IL-1β) were detected by Western blotting. The levels of reactive oxygen species (ROS) were detected by dihydroethidium (DHE) staining. Results: Compared with the C group, the expression level of cardiac Nox4 protein was increased (P<0.01), the level of MDA, activity of LDH and the level of ROS were increased significantly (P<0.01), and the expressions of TNF-α and IL-1β protein were augmented in the heart of rats with MI (P<0.01). However, the expressions of SIRT1 mRNA and protein and the activity of SOD were obviously decreased in MI group (P<0.01). Furthermore, compared with the MI group, AIT increased the expressions of SIRT1 mRNA and protein and the activity of SOD in the heart of ME group (P<0.01); Meanwhile, the expressions of cardiac Nox4, MDA level, LDH activity and ROS level were diminished in ME group (P<0.01) as well as the decreased expressions of TNF-α and IL-1β protein (P<0.01). SIRT1 expression was negatively related to the expressions of NOX4 and ROS. Conclusion: AIT obviously inhibited myocardial oxidative stress and inflammatory reaction, improved cardiac function in rats with MI, and the mechanism was closely related to the activation of SIRT1-Nox4-ROS signaling pathway.
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TECHNICAL NOTES
A method of marking neurons on brain slices using patch clamp technique
GAO Jie, HUANG Yan-yun, ZHANG Yu-tong, DU Xiang-xin, ZHANG Li-na, HAO Na, GUO Xia, LI Jian-guo, ZHANG Yu
CJAP. 2021, 37 (4): 445-448.   DOI: 10.12047/j.cjap.6035.2021.031
Abstract   PDF (1151KB)
Objective: To introduce a method of marking neurons using patch clamp technique. Methods: The brain slices of the target area was cut with a vibrating microtome. The glass microelectrode was perfused with the electrode liquid containing NeurobiotinTM Tracer, and the whole-cell patch-clamp recording was performed. After recording, the brain slices were fixed and rinsed with 4% paraformaldehyde. After stained in phosphate buffer with Streptavidin-Texas Red and Triton X-100 for at least 2 hours, the neurons can be observed under a fluorescence microscope. Results: The cell membrane voltage was clamped at -70 mV, and the neuron showed a gradually increasing membrane current after step stimulation. When recording in the current clamp mode, the step stimulus caused the neuron to depolarize to the threshold potential and then burst into action potentials. The morphology of intact neurons with clear cell body and protrusions of a neuron could be observed under a fluorescence microscope. Conclusion: This method is suitable for observing the morphological features of the recorded neuron after patch clamp experiments, which is easy to operate, and the image is intuitive and clear.
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2021
Vol.37
No.3 
2021-05-28
pp.225-336
No.2
2021-03-28
pp.113-224
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2021-01-28
pp.1-112
2020
Vol.36
No.6 
2020-11-28
pp.529-672
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2020-09-28
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2020-07-28
pp.289-384
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2020-05-28
pp.193-288
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2020-03-28
pp.101-192
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2020-01-28
pp.1-96
2019
Vol.35
No.6 
2019-11-28
pp.481-576
No.5
2019-09-28
pp.385-480
No.4
2019-07-28
pp.289-384
No.3
2019-05-28
pp.193-288
No.2
2019-03-28
pp.101-192
No.1
2019-01-28
pp.1-96
2018
Vol.34
No.6 
2018-11-28
pp.481-576
No.5
2018-09-28
pp.385-480
No.4
2018-07-28
pp.289-384
No.3
2018-05-28
pp.193-288
No.2
2018-03-28
pp.97-192
No.1
2018-01-28
pp.1-96
2017
Vol.33
No.6 
2017-11-28
pp.481-576
No.5
2017-09-28
pp.385-480
No.4
2017-07-28
pp.289-384
No.3
2017-05-28
pp.193-288
No.2
2017-03-28
pp.97-192
No.1
2017-01-28
pp.1-96
2016
Vol.32
No.6 
2016-11-28
pp.481-576
No.5
2016-09-28
pp.385-480
No.4
2016-07-28
pp.289-384
No.3
2016-05-28
pp.193-288
No.2
2016-03-28
pp.97-192
No.1
2016-01-28
pp.1-96


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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
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Editorial Board,Chinese Journal of Applide Physiology;Dali Dao,Tinanjin 300050,China



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