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  • Table of Content
      28 September 2016, Volume 32 Issue 5 Previous Issue    Next Issue
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    Effects of strong noise on cognitive function and some biochemical indexes in serum of guinea pigs
    ZHAO Xiao-yu, LIU Shu-hong, ZHAO Tong, ZHU Ling-ling, FAN Ming, ZHAO Yong-qi, FANG Tao
    CJAP. 2016, 32 (5): 385-389.   DOI: 10.13459/j.cnki.cjap.2016.05.001
    Abstract   PDF (978KB) ( 12 )
    Objective: To explore the effect of strong noise on cognitive function and stress hormones, biochemical metabolism enzymes, neuropeptide etc in theserum of guinea pigs, and screen the sensitive bio-markers of noise-induced cognitive impairment.Methods: Twenty-four guinea pigs were randomly divided into the noise exposure group and the control group, after the white noise at 120 dB continuously exposed for 4 h. Using Morris water maze to detect the cognitive ability of guinea pigs, and using Elisa kits to detect the content of neuropeptide Y (NPY), neuropeptide P (NPP), neuroglobin (NGB), c-reactive protein (CRP), cortisol (CORT), adrenocorticotropic hormone (ACTH), corticotropin releasing hormone (CRH) and epinephrine (E), norepinephrine (NE), angiotensin Ⅱ (AngⅡ), aldosterone (ALD), oxytocin (OT), vasopressin (VAP), alanine aminotransferase (ALT), aspartate transaminase (AST), lactate dehydrogenase (LDH), and creatine kinase (CK) in the serum of guinea pig. Multiple regression analysis was used to evaluatethe sensitive indexes for the influence of cognitive function.Results: Morris water maze test indicated that the average escape latency of noise exposure group extended obviously, cross platform number and residence time in target quadrant were significantly reduced compared with control group(P<0.01). Elisa results showed that the content of the biochemical indexes above of noise exposure grouphad significant statistical difference except AngⅡ compared with control group(P<0.01), among them, the content of CORT, NPY, NE, NGB, NPP, CK changed obviously and contributed greatly to cognitive in turn.Conclusion: Strong noise at 120dB continuous exposure for 4 h caused a significant decline on cognitive function of guinea pigs, it might be related to the abnormal levels of the physiological and biochemical indexes in serum such as stress hormones, biochemical metabolism enzyme, neuropeptides, neuroglobin etc, and we preliminarily filtrated several sensitive index associated with noise-induced cognitive impairment, including CORT、NPY、NE、NGB、NPP and CK.
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    Establishment and evaluation of oxidative stress injury model of in vitro rat aortic endothelial cells
    WU Lei, ZHOU Xue-si, GAO Xiu-jie, YANG Miao-miao, ZHANG Zhi-qing, LI Jing-gang, WANG Tian-hui
    CJAP. 2016, 32 (5): 390-394.   DOI: 10.13459/j.cnki.cjap.2016.05.002
    Abstract   PDF (978KB) ( 13 )
    Objective: To establish a model of oxidative stress injury in cultured rat aortic endothelial cells, and to provide a basis for the research of cell injury and apoptosis. Methods: The rats were decapitated to get the aorta in thoracic operation under aseptic conditions. By subculture after tissue block culture method to get sufficient aortic endothelial cells, cultured in 96-well plates or grow on cover glass for the following test. Without H2O2 group as a control group, with different doses of H2O2 (100,200,300,400,500 μmol/L) treated endothelialcells in 12 h to screen the optimal dose. Based on the results, with the same dose of H2O2 (100 or 200 μmol/L) acted on endothelial cells respectively in different time (3, 6, 9, 12 and 24 h) to screen the optimal duration. Each group was made in sextuplicate. The establishment of the model was evaluated by immunofluorescence,cell viability testing, biochemical indicators detection (lactate dehydrogenase(LDH), nitric oxide(NO), malondialdehyde(MDA), superoxidedismutase(SOD))and apoptosis index testing. Results: Endothelial cells were cultured successfully and verified by immunofluorescence staining of intracellular antigen Ⅷ collagen. With the increase of H2O2 doses at the same action time 12 h, the cell viability was significantly decreased (77.63%±5.20% to 40.90%±2.10%). The same dose(100 μmol/L group and 200 μmol/L group)with the action time increasing, the cellviability was significantly decreased (100 μmol/L group was 86.83%±12.11% to 44.26%±5.70%, 200 μmol/L group was 78.28%±11.98% to 34.45%±5.87%). At dose of H2O2 was 100 μmol/L and treated in 3,6,9,12 and 24 h, LDH-L and MDA were significantly increased after 9 h while NO and SOD were significantly decreased. In H2O2 dose of 100 μmol/L and action time 12 h, flow cytometry showed endothelial cellapoptosis rate was 16.92%±2.37%, significantly higher than the control group of 2.68%±0.47%(P<0.01); TUNEL detected endothelial cell apoptosis index was17.65%±2.36%, which was significantly higher than that in the control group of 3.23%±0.57%(P<0.01). Conclusion: The method was successfullyestablished a model of oxidative stress injury in cultured rat aortic endothelial cells, explore the moderate conditions that induced cells injury and apoptosis which could be a basis for the research.
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    The experimental study of simvastatin on improving aspirin resistance in diabetic rats
    HAO Wei-jun, KE Se-zhang, LIU Lin, LI Jian-hua, LUO Xiao-xing, SUN Yu-fa, CAO Jian, FAN Li
    CJAP. 2016, 32 (5): 395-400.   DOI: 10.13459/j.cnki.cjap.2016.05.003
    Abstract   PDF (1233KB) ( 8 )
    Objective: The purpose of the present study was to investigate the effects of the combination of aspirin, simvastatin in diabetic rat on platelet function.Methods: Eight-week-old male Wistar rats were selected and randomly divided into diabetic group (n=48) and normal control group (n=48). Diabetic rats were injected with 1% STZ (65mg/kg, dissolved in 0.l mmol/L, pH 4.5 citrate buffer) to induce diabetic model and the rats in normal control group were injected with the same dose of citrate buffer. A rat with blood glucose greater than 16.8 mmol/L and along with diabetic symptoms of polydipsia, polyuria and weight loss was considered the successful model of diabetes. Diabetic rats and normal Wistar rats were randomly divided into 4 groups and given aspirin(10 mg/kg), simvastatin(2 mg/kg), combination of aspirin(10mg/kg) and simvastatin(2 mg/kg), PBS for 8 weeks, respectively. The platelet function and the expression of CD62P were evaluated. The levels of nitric oxide (NO), endothelin (ET), thromboxane B2(TXB2), prostacyclin (PGI2), adiponectin (APN), TXB2 were detected in the serum. The expressions of heme oxygenase-1(HO-1), HO-2, endothelial nitric oxide synthase(e-NOS), p-eNOS, B-cell lymphoma-2(Bcl-2), cyclooxygenase-2(COX-2) in thoracic aorta were evaluated by Western blot.Results: Compared with control rats, diabetic rats had high platelet aggregation and activation (P<0.05), which treated aspirin also showed lower aspirin sensitivity (P<0.05). The combination of drugs upregulated the expression of HO-1, eNOS, p-eNOS, BCL-2, APN levels and decreased the expression of COX-2, and had a greater inhibitory effect on platelet aggregation and activation. The combination of drugs improved endothelial function, adjusted TXA2/PGI2 levels and increased NO levels, which resulted in a great potential antiplatelet effect.Conclusion: These results suggest that simvastatin may improvethe effect of aspirin on anti-platelet function in diabetic rats.
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    The impact of electrophysiology of central dopaminergic neurons and the depression-state induced by chronic neuropathic pain
    FU Bo, WENG Xie-chuan, WANG Jing, HUANG Tao, WANG Bin, LIN Shi-de, LIU Shao-jun
    CJAP. 2016, 32 (5): 403-407.   DOI: 10.13459/j.cnki.cjap.2016.05.005
    Abstract   PDF (1811KB) ( 7 )
    Objective: To explore the underlying electrophysiological mechanism of depression induced by chronic pain in dopaminergic neurons in midbrain ventral tegmental area (VTA) of rats. Methods: Twenty-four healthy adult rats were divided into two groups randomly(n=12):12 rats formed the sham group by exposing the spared nerve, and another 12 rats were served as the spared nerve injury (SNI)group by branchedness sciatic nerve injury surgery. The mechanical allodynia test were detected on the day of 3, 7, 14, 28, 42 and 56 after surgery, and the depressive-like behaviors such as open-field test, sucrose preference and forcedswim test were detected at the same time. Then we used the Multichannel Acquisition Processor (MAP) system to record the firing activity of neurons in VTA in both Sham rats and SNI rats.Results: ①Comparing to sham rats, the paw withdrawalmechanical threshold of SNI rats was decreased significantly (P< 0.01);② According to depression-related behavioral test, SNI rats showed significant difference in open field text, sucrose preference, focus swim text comparing with Sham rats (P< 0.01);③ The firing rate and burst activity of dopaminergic neuronsin midbrain VTA are increased in depression rats compare to sham rats(P<0.05).Conclusion: Chronic pain could induce depression, and the increase of spontaneous firing rate of dopaminergic neurons in midbrain VTA might be contribute to the depression induced by the chronic neuropathic pain.
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    The effects and mechanisms of ligustrazine injection on pulmonary arterial hypertension in COPD patients
    ZHAO Mei-ping, MIU Chu-lei, ZHANG Cong-cong, ZHENG Meng-xiao, HUANG Lin-jing, WU Cheng-yun, WANG Wan-tie
    CJAP. 2016, 32 (5): 408-412.   DOI: 10.13459/j.cnki.cjap.2016.05.006
    Abstract   PDF (1268KB) ( 20 )
    Objective: To observe the effects of ligustrazine hydrochloride injection(LHI) on pulmonary arterial hypertension in chronic obstructive pulmonary disease(COPD) patients and to investigate its possible mechanisms. Methods: Twenty-two cases of patients with COPD were randomly divided into conventional treatmentgroup (group C) and ligustrazine treatment group(group L), 11 persons were randomly selected from healthy subjects without lung disease served as normal control group(group N). Group C was given bed rest, low flow oxygen inhalation, bronchial diastolic agent, glucocorticoid and antibiotics and other conventional treatment, and group L was added with ligustrazine hydrochloride injection on the above mentioned basis treatment, group N was given no treatment. After 2 weeks, lung function, blood gas analysis and pulmonary arterial pressure were compared among the three groups, and the content of H2S in plasma was tested with sensitive sulfur electrode method. Results: ①After two weeks treatment, in group L and group C pulmonary function, blood gas analysis, pulmonary artery pressure were obviously improved, and group L was better than group C (P<0.05); ② In group L the content of H2S was increased (P<0.01), group C had no significant difference (P>0.05), and there was a significant difference between the two groups (P<0.01). Conclusion: Combination with LHI can effectively improve lung function. LHI mayrelieve hypoxic hypercapnia pulmonary hypertension induced by COPD through raising the content of H2S.
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    Effects of fasudil on isoproterenol induced cardiac hypertrophy in rats
    CHENG Jie-kun, SUN Xiao-hui, GAO Li-ping, LI Le
    CJAP. 2016, 32 (5): 414-418.   DOI: 10.13459/j.cnki.cjap.2016.05.008
    Abstract   PDF (1093KB) ( 15 )
    Objective: To investigate the effects and mechanisms of fasudil (Fas) on cardiac hypertrophy (CH) induced by isoprenaline(Iso) in rats. Methods: Except for the normal control group, the rest three groups rats were injected subcutaneously with Iso(5mg/kg) for setting up CH model. SD rats were randomly divided into four groups:Normal control group, Iso model group, Fas with low-dose group (5 mg/kg, i.p) and Fas with high-dose group (20 mg/kg, i.p). Animals were treated with Fas for 8 weeks. After the treatment, the following index were detected:heart rate (HR), left ventricular systolic pressure (LVSP), left ventricular end diastolic pressure (LVEDP), left ventricular systolic maximum rate (+dp/dtmax) and left ventricular diastolic maximum rate (-dp/dtmax). The body weight(BW) and heart weight (HW) were weighed and heart weight index (HWI) was calculateed. Myocardial tissue specimens, HE and Masson staining were performed for observing the histopathological changes. The protein expression of extracellular signal-regulated kinase 1/2 (ERK1/2) in myocardial tissue was determined by using immunohistochemical methods. The expression of ERK1/2 mRNA in the myocardial tissue was detected by using semi-quantitative RT-PCR method. Results: Compared with normal control group, HR and LVEDP wereincreased significantly, while LVSP and ±dp/dtmax were decreased significantly; HWI was increased significantly in CH group; The myocardial cell volume and the cell gap were increased,cells arranged disorder, and severe myocardial interstitial fibrosis was presented in CH group. The expression of ERK1/2 mRNA was significantly increased. After Fas treatment, the systolic and diastolic functions of heart were improved, cell volume was reduced, cell gap became small, cells arranged in neat rows, and the fibrosis was significantly reduced. The expressions of RhoA, ERK1/2 mRNA were significantly reduced. The damages of myocardial tissue were improved in different degrees. Conclusion: ERK1/2 signaling pathway is involved in Iso induced CH. Fas protects against the CH of rats induced by Iso, which mechanisms may be related with blocking ERK1/2 signaling pathway.
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    Effects of Mongolian new medicine-Ⅱ on cardiac functions, endoplasmic reticulum stress and apoptosis in congestive heart failure rats with dilated cardiomyopathy
    CHEN Shao-qing, WANG Yi-lin, CAO Xian-yu, ZHANG Qing-shan, CHAI Hua, TAO Xie-xin, ZHAO Ming
    CJAP. 2016, 32 (5): 419-423.   DOI: 10.13459/j.cnki.cjap.2016.05.009
    Abstract   PDF (1254KB) ( 10 )
    Objective: To investigate the effects of Mongolian new medicine-Ⅱ oncardiac functions, myocardial pathology, endoplasmic reticulum stress and myocardial apoptosis in congestive heart failure with dilated cardiomyopathy in rats.Methods: Thirty SD male rats were randomly dividedinto 3 groups(n=10):control group, dilated cardiomyopathy group (intraperitoneal injection of adriamycin 2 mg/kg body weight, 1 time/week, 4 weeks after treatment were observed for 4 weeks), Mongolian new medicine-Ⅱ group(intraperitoneal injection of adriamycin 2 mg/kg body weight, 1 time/week, 4 weeks after treatment, 30 mg/(kg·d)was given Mongolian new medicine-Ⅱ orally for 4 weeks). During the experiment, general conditions of rats were observed. After 8 weeks, these rats were killed after measurement of the cardiac function indexes by high frequency echocardiography. The morphological changes of myocardial tissues were observed by using HEstaining, VG staining and electron microscopic. The myocardial apoptosis was detected by TUNEL method and the expressions of endoplasmic reticulum chaperone GRP78, GRP94, pro-apoptotic factor CHOP and caspase-3 were monitored by Western blot. Results: ① Compared with dilated cardiomyopathy group, the cardiac systolic and diastolic functions were significantly increased in Mongolian new medicine-Ⅱ group, which were reflected in that left ventricular contraction diameter(LVIDs) and left ventricular end-diastolic diameter(LVIDd) were decreased, and left ventricular shortening fraction(FS) and ejection fraction(EF) were increased. The hemodynamic parameters of rats were improved significantly in Mongolian new medicine-Ⅱ group. ②Compared with dilated cardiomyopathy group, the myocardial lesion score was decreased and fibrosis of tissue space was relieved in Mongolian new medicine-Ⅱ group. ③Compared with dilated cardiomyopathy group, the apoptosis of myocardial cells was decreased. ④The expressions of endoplasmic reticulum chaperone GRP78, GRP94, pro-apoptotic factor chop and caspase-3 were decreased in Mongolian new medicine-Ⅱ group. Conclusion: Mongolian new medicine-Ⅱ could improve the pathologic alterations of cardiac cells and cardiac functions, decrease endoplasmic reticulum stress, the degree of fibrosis and myocardial apoptosis. The experimental results may be one of the mechanisms of treatment function of Mongolian new medicine-Ⅱ on dilated cardiomyopathy.
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    Effects of exendin-4 on methylglyoxal-induced oxidative stress in PC12 cells
    ZHOU Qing, WANG Yan-ping, LIU Xiao-ying, LIU Xiao-hong, PAN Xiao-dong, CHEN Zhou, LIU Li-bin
    CJAP. 2016, 32 (5): 426-430.   DOI: 10.13459/j.cnki.cjap.2016.05.011
    Abstract   PDF (1056KB) ( 11 )
    Objective: To study whether Exendin-4(Ex-4) could influence oxidative stress in PC12 cells induced by methylglyoxal and its underlying mechanism. Methods: PC12 cells were cultured with methylglyoxal (0,0.25,0.50,0.75,1.0,2.0 mmol/L) for 12~48 h, or PC12 cells were pretreated with Ex-4 (25, 50, 100, 200 nmol/L) for 24 h then incubatedwith methylglyoxal (0.75 mmol/L) for 24 h. MTT assay was used to measure cell viability. Fluorescent probe method was used to detect reactive oxygen species (ROS) expression. Xanthine oxidase method was used to detect superoxide dismutase (SOD)activity. With pretreatment of Exendin-4 (100 nmol/L) for 24 h,the expressions ofP-IκBα, Inhibitor of NF-κB-α IκBα were detected by Western blot after PC12 cells were exposed to methylglyoxal (0.75 mmol/L) for 1 h. Results: Following methylglyoxal administration, cell viability was gradually decreased in a dose-and time-dependent manner. Pretreatment with Ex-4 for 24 hours, cellviability were gradually increased compared with methylglyoxal-alone group. Pretreatment with Ex-4 (100 nmol/L) for 24 hours, ROS expression was reduced by65.30% (P<0.01) compared with methylglyoxal-alone group, ROS expression in NAC-pretreatment group was reduced by 107.40% (P<0.01); SOD activity in the Ex-4 pretreatment group was increased by 5.30 U/mg prot (P<0.01), SOD activity in the NAC pretreatment group was increased by 8.53 U/mg prot (P<0.01);the ratio of P-IκB-α/IκB-α in the Ex-4 pretreatment group was reduced by 25.50% (P<0.01), the ratio of P-IκB-α/IκB-α in the NAC pretreatment group was reduced by 35.14% (P<0.01). Conclusion: This study demonstrates that Ex-4 can increase the viabilities of PC12 cells and protect PC12 cells from oxidative stress induced by methylglyoxal, the mechanism may involve in suppressing the activation of protein IκB-α.
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    Changes of theta oscillation of prelimbic cortex in heroin-induced CPP rats
    YE Zheng, YU Ran, LI Jing, LI Min, ZHU Zai-man, WANG Dan-dan, PAN Qun-wan
    CJAP. 2016, 32 (5): 431-437.   DOI: 10.13459/j.cnki.cjap.2016.05.012
    Abstract   PDF (1696KB) ( 12 )
    Objective: To investigate the relationship between the changes of neural oscillations and the drug-seeking motivation,record the telemetric local field potentials (LFPs) of the prelimbic cortex (PrL) in the latency of drug-seeking behavior of conditioned place preference (CPP) rats induced by heroin. Methods: The recording electrode was stereotactically implanted intothe PrL cortex of rats. The animals were then randomly divided into operation-only control and heroin-induced CPP groups, respectively. A CPP video system in combination with awireless telemetry device was used for recording LFPs when the rats shuttled between black-white chamber for drug-seeking. The LFPs were analyzed by fast Fourier transform (FFT) and wavelet packet extraction. Results: Compared with operation-only control group, the LFPs recorded in PrL area of heroin-induced CPP group of rat during black-white chamber shuttling showed that the percentage of θ rhythm were increased in right and left PrL, the percentage of γ3 rhythm was increased in left PrL, the phase-amplitude coupling of θ and γ3 was increased in left PrL. After MK-801 Maleate micro injection to PrL area,drug-seeking behavior of rat was markedly reduced and the percentage of θ oscillation was depressed,the phase-amplitude coupling of θ and γ3 was depressed in left PrL compared with operation-only control group. Conclusion: These results suggest that increase of θ oscillations of PrL area may be related to the drug-seeking motivation and behavior launching in heroin-induced CPP groups of rat. The changes of θ oscillations also have close relationship with glutamatergic neuron and the receptor of it on PrL area.
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    Comparative study of high altitude chronic hypoxia on renal function in yak and migrated cattle on Qinghai-Tibetan plateau
    LI Yu-xian, LIU Feng-yun, HU Lin, LIU Shi-ming, WU Tian-yi
    CJAP. 2016, 32 (5): 440-443.   DOI: 10.13459/j.cnki.cjap.2016.05.014
    Abstract   PDF (967KB) ( 23 )
    Objective: To investigate the difference of renal function among yak, migrated cattle on Qinghai-tibetan plateau and lowland cattle, and to further explore the characteristics of renal to adapt chronic hypoxia.Methods: The serum samples of yak(n=84) were collected at 3 000 m,3 500 m,4 000 m and 4 300 m respectively,meanwhile the serum samples of migrated cattle on plateau (n=22) and lowland cattle (n=39) were also collected.The levels of blood urea nitrogen(BUN), creatinine(Crea), blood urea nitrogen/creatinine(BUN/Cr), uric acid(UA), carbon dioxide binding rate(CO2cp), glucose(GLU) in serum were measured by using fully automatic blood biochemical analyzer. We analysed the differences among the above renal functions.Results: With the altitudeincreased, the results showed the levels of UA and CO2cp of yak were increased significantly, as compared to cattle, the levels of BUN and BUN/Cr were increased significantly compared with migrated cattle on plateau and lowland cattle, thelevels of CO2cp and GLU were decreased significantly compared with lowland cattle. As compared to migrated cattle on plateau, the levels of BUN and BUN/Cr of lowland cattle were decreased significantly, the levels of UA and CO2cp were increased significantly.Conclusion: The results indicated that theyak were adaptedto the plateau hypoxia environment and migrated cattle maybe not adapt to the low oxygen environment, they were under the stress situation.
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    Effect of DaChaiHu Decoction on pancreatic fibrosis induced by DBTC combined with alcohol and the mechanism of TGF-beta/Smad signaling pathway
    XU Xiao-fan, JIANG Ting-ting, LIU Fang, ZHANG Xiao-qin, SHI Ying-li, CHEN Yu, LI Tao, ZHANG Hong
    CJAP. 2016, 32 (5): 444-448.   DOI: 10.13459/j.cnki.cjap.2016.05.015
    Abstract   PDF (1127KB) ( 16 )
    Objective: To investigate the effects of transforming growth factor-β (TGF-β) and Smad in chronic pancreatitis (CP) induced by dibutyltin dichloride (DBTC) combined with ethanol, and the effect for prevention and treatment of pancreatic fibrosis by DaChaiHu Decoction (tradeiional Chinese medicine). Methods: Ninety-six healthy male Kunming mice were randomly divided into control group (Con group), chronic pancreatitis (CP group) group, Dachai Hu decoction treatment group (DCHD group) (n=32). Then the mice were treated with DBTC 8 mg/kg by tail vein injection and fed with 10% ethanol replacing the normal drinking water to replicate mouse CP model. After three days of modeling the mice were randomly divided into CP group and DCHD group. The mice in DCHD group were administered intragastrically with DaChaiHu Decoction (1 g/ml, 6 g/kg·d) at the following 8 weeks. After modeling for 1 week, 2 weeks, 4 weeks and 8 weeks, the mice were anesthetized and sacrificed(n=8). The morphology and the degree of fibrosischanges of pancreatic tissue were detected by HE staining. The protein expressions of type I collagen and TGF beta R I, p-Smad 2/3 and Smad 7 in pancreatic tissue were detected by Western blot assay. The expressions of MMP-1/TIMP-1 mRNA inpancreatic tissue were detected by RT-PCR.Results: Compared with control group, DBTC combined with ethanol induced CP with increased activity of serum amylase and hyaluronic acid level. While in DCHD group, the activity of amylase and hyaluronic acid level were decreased significantly. Compared with control group, the protein expressions of COLA1, TGF beta R I, p-Smad 2/3 in CP Group were elevated,but the expression of Smad 7 was significantly reduced; In DCHD treatment group,the expression of TGF beta R I, p-Smad 2/3 and COLA1 were reduced, and the protein expression of Smad 7 was increased. In 2w and 4w, the level of MMP-1mRNA continued to decrease, while TIMP-1mRNA expression was increased significantly (P<0.01). At each time point of DCHD group, the expression of MMP-1 were markedly increased and TIMP-1 were reduced, compared with those of the CP group (P<0.05). Conclusion: DaChaiHu Decoction play a role in the prevention and treatment of chronic pancreatitis and the development of fibrosis, the mechanism may related to inhibit the activation of TGF beta/Smad signaling pathway, and regulate the balance between synthesis and degradation of extracellular matrix.
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    The roles and mechanisms of p-STAT3 signaling pathway in acute pancreatitis
    SHI Ying-li, LIU Fang, ZHANG Xiao-qin, JIA Xiao-yun, LI Tao, XU Xiao-fan, ZHANG Hong
    CJAP. 2016, 32 (5): 450-453.   DOI: 10.13459/j.cnki.cjap.2016.05.017
    Abstract   PDF (1024KB) ( 13 )
    Objective: To detect the expression ofsignal transducer and activator of transcription 3 (STAT3) in pancreatic tissue of the mouse model of pancreatitis, and to explore its role in the evolution of acute pancreatitis.Methods: Forty-eight healthy male balb/c mice were randomly divided into 3 groups (n=16):control group (Con) 0.09% NaCl, intraperitoneal injection; mild acute pancreatitis group (MAP) caerulein, intraperitoneal injection; severe acute pancreatitis group (SAP) caerulein plus lipopolysaccharide(LPS), intraperitoneal injection. The mice were sacrificed after 2 h and 6 h after intraperitoneall injection. Serum was isolated for amylase activity. Pancreatic was isolated and weighedto calculate the pancreatic wet weight ratio. Myeloperoxidase (MPO) activity was measured to assess the degree of inflammatory cell infiltration in lung tissue. Using HE staining, the pathological changes of pancreatic and lung were observed under the light microscope. The expression of phosphorylated STAT3 (p-STAT3) was detected by Western blot.Results: Compared with control group, serum amylase activity, pancreatic wet weight ratio and lung MPO activity were significantlyincreased (P<0.05) in MAP and SAP group at each time point, especially SAP group showed higher levels of MPO activity than that in MAP group (P<0.01). The pathological changes of pancreas and lung were observed after modeling in 2 h. Western blot showed the expression of p-STAT3 could be detected in SAP group, the level increased most significantly after modeling 2 h, and decreased slightly after 6 h. The level of p-STAT3 was low in MAP group and negative in Con group at each time point.Conclusion: The expression of p-STAT3 in MAP and SAP groups are significantly different from that in control group, which indicates that STAT3 isclosely related in acute pancreatitis. Inhibition of STAT3 activity is a potential target to alleviate acute pancreatitis progression.
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    The expression of calcium sensing receptor(CaSR) and MAPK pathway changes in myocardial cells of epilepsy rats
    GUO Jin, LIU Yang, WANG Chao, BAI Ling-ling, HAN Xiao-wei, ZHANG Xin-yang, SUN Yan-qiu, WANG Lu-chuan, JIANG Zhi-mei
    CJAP. 2016, 32 (5): 454-458.   DOI: 10.13459/j.cnki.cjap.2016.05.018
    Abstract   PDF (1708KB) ( 13 )
    Objective: To investigate the changes of the myocardial cells in chronic epileptic rat model and to observe the expression of calcium sensing receptor(CaSR) and mitogen-activated proteinkinase(MAPK)pathway changes in epilepsy rats. Methods: The chronic epileptic rat model was induced bypentetrazole (PTZ). Adult male Wistar rats were divided into 5 groups randomly, and there were 12 rats in each group. The rats in model group were treated with a sub-convulsivedose of PTZ (35 mg/kg) by intraperitoneal injection for 28 d. After stopping a week, the same dose of PTZ test was conducted. The control group was treated with isovolumetric saline instead of PTZ by intraperitoneal injection. According to Racine behavior grading standards the rat emerged two levels above epileptic seizure 5 consecutive times, which was considered the chronic epilepsy model successful ignition. The intervention factors included spermine(calcium-sensing receptor agonist, 3 μmol/L) and Chalhex231(calcium-sensing receptor inhibitor, 2 μmol/L). The serum creatine kinase (CK) and creatine kinase isoenzyme(CK-MB)were detected. The cardiac functions, morphological changes of rat myocardial tissue, myocardial cell ultrastructure, myocardial cell calcium sensing receptor and extracellular regulated protein kinase (ERK), p-ERK, p-JNK expression were carried out. Results: Compared with normal control group, CK, CK-MB inPTZ group were increased obviously. The cardiac compliance and left ventricular function were decreased, E/A<1 by echocardiography. The myocardial ultrastructure showed serious injury. The expressions of CaSR and p-JNK were increased, but the expression of p-ERKwas decreased. Spermine could promote the expressions of CaSR and p-JNK, and decrease the expression of p-ERK in epilepsy; however, the role of Chalhex231 wasopposite. Conclusion: The level of CaSR expression increased in chronic epileptic rat model. CaSR activated the expressions of MAPK of the myocardial cells,andthen influenced the cardiac myocyte apoptosis.
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    The protective effects of dexmedetomidine on perioperative myocardial injury in patients with hypertensive myocardial hypertrophy
    WU Yong, ZHANG Xiao-bao, QIAN Yan-ning, XIAO Hang
    CJAP. 2016, 32 (5): 459-462.   DOI: 10.13459/j.cnki.cjap.2016.05.019
    Abstract   PDF (968KB) ( 12 )
    Objective: The purpose of this study was to evaluate the effects of dexmedetomidine (DEX) on patients with hypertensive myocardial hypertrophy.Methods: Fifty four patients with hypertensive myocardial hypertrophy were enrolled in the study and were randomly divided into two groups (n=27). Patients in groupD were pretreated with DEX (1 μg/kg) before induction and then maintain with 0.5 μg/(kg·h) DEX. Patients in group C were pretreated with saline at the same time. All patients were connected with holter recorder 2 h before anesthesia and were continuously recorded for 24 h. Blood sample were collected to measure ischemia modified albumin(IMA) and serum cardiac troponin I (cTnI) at the time of T0 (before induction), T1(1 h after surgery), T2(4 h after surgery), T3(12 h after surgery) and T4(24 h after surgery). The surgery time, blood loss and side effect of two groups were recorded at the same time.Results: The serum IMA level in group D was lower than that of group C at the time of T1, T2 and T3 (P<0.05). The serum cTnI in group C was higher than that of group D at the time of T1, T2, T3 and T4 (P<0.05). Changes of ST and complicated ventricular arrhythmias ingroup D were lower than those of group C (P<0.05).Conclusion: DEX could reduce the incidence of myocardial damage, changes of ST and complicated ventricular arrhythmias in patients with hypertensive myocardial hypertrophy.
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    Changes of lipid levels in mice with hypoxic pulmonary arterial hypertension
    FAN Feng-yun, SHEN Wan-ting, ZHENG Qing-qing, GAO Yuan, HU Kai-yuan, FAN Xiao-fang, MAO Sun-zhong, GONG Yong-sheng
    CJAP. 2016, 32 (5): 463-465.   DOI: 10.13459/j.cnki.cjap.2016.05.020
    Abstract   PDF (994KB) ( 12 )
    Objective: To observe the changes of lipid levels in mice with pulmonary hypertension induced by hypoxia. Methods: The animal model of hypoxic pulmonary hypertension was established by exposing the mice to isobaric hypoxic chamberfor 3 weeks (23 h/d, regular chow feed). Twenty male C57BL/6 mice were randomlydivided into normoxia group and hypoxia group (n=10). The concentrations of total cholesterol, low density lipoprotein (LDL) and high density lipoprotein (HDL) in plasma were detected by Elisa method.The mRNA levels of HMG-CoAreductase (HMGCR), low density lipoprotein receptor (LDLR), scavenger receptor class B1 (SR-B1), and sterol regulatory element-binding factor-2 (SREBF2) in liverwere measured by real-time PCR. Results: ① The right ventricular systolic pressure (RVSP) and the weight ratio of right ventricle (RV) to left ventricle plus septum (LV+S) of hypoxia group were significantly higher than those of normoxia group (P<0.05).② The concentrations of HDL and HDL/LDL in plasma were significantly higher in hypoxia group, compared with normoxia group (P<0.05).③The mRNA levels of LDLR and SR-B1in liver were significantly down-regulated in hypoxia group(P<0.05).④RVSP were significantly negative correlated with HDL/LDL, the gene expression of LDLR and SR-B1 (P<0.05). Conclusion: Abnormal lipid metabolism participates in the pathological proceeding of pulmonary hypertension induced by hypoxia.
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    Electrophysiological effects of ischemia/reperfusion (I/R) on pacemaker cells in guinea-pig left ventricular outflow tract and the effects of antiarrhythmic drugs treatment
    ZHAO Lan-ping, XUE Shu-fang, CHEN Yan-jing, WANG Xue-fang, CHEN Li-feng, FAN Yu-hong, ZHANG Xiang-yu
    CJAP. 2016, 32 (5): 466-470.   DOI: 10.13459/j.cnki.cjap.2016.05.021
    Abstract   PDF (1326KB) ( 11 )
    Objective: To investigate the electrophysiological effects of ischemia/reperfusion (I/R) on the spontaneous slow response action potentials in guinea-pig left ventricular outflow tract and the effects of antiarrhythmic drugs onI/R. Methods: The action potentials of pacemaker cells in guinea-pig left ventricular outflow tract were recorded by conventional intracellular microelectrode technique. The influences of ischemia(I) 10 min, reperfusion(R) 2 min, and R 15min on the spontaneous slow response potentials were investigated. The effects of lidocaine, propafenone, amiodarone, verapamil, adenosine, and sodium nitroprusside (SNP) on I/R were also studied. Electrophysiological parameters were examined:velocity of diastolic depolarization(VDD), rate of pacemaker firing(RPF), maximal diastolic potential(MDP), maximal rate of depolarization(Vmax), amplitude of action potential(APA), 50% and 90% of duration of action potential(APD50 and APD90). Results: ①In I 10 min group, the values of VDD, RPF, Vmax and APA were decreased markedly compared with control group (P<0.05, P<0.01).In R 2 min group, VDD and RPF were increased significantly(P<0.01), MDP was increased notably(P<0.05), APD50 and APD90 were shortened significantly compared with I 10 min and control group. Vmax was increased markedly vs control group(P<0.05). APA was decreased notably vs I 10 min group (P<0.05), but was increased markedly vs control group(P<0.05). In R 15 min group, the action potentials recovered gradually to the levels of control group. ② Compared with I 10 min/R 2 min group, 1 μmol/L lidocaine, 10 μmol/L propafenone, 1 μmol/L amiodarone, 1 μmol/L verapamil, 50 μmol/L adenosine, and 10 μmol/L SNP decreased VDD and RPF significantly. Conclusion: I/R injury can trigger abnormal spontaneous activities of guinea-pig left ventricular outflow tract.The electrophysiological effects of I/R injury on left ventricular outflow tract can be treated by antiarrhythmic drugs.
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    Construction of eukaryotic expressing vector of human P2X7 and establishment of stable transfectant cell line
    WEI Lin-yu, LU Na, MENG Li, LI Xin-juan, LI Lu, LI Chao-kun, LI Dong-liang
    CJAP. 2016, 32 (5): 471-475.   DOI: 10.13459/j.cnki.cjap.2016.05.022
    Abstract   PDF (1425KB) ( 21 )
    Objective: To construct eukaryotic expression vector of human P2X7gene and transfect HEK293 cells so as to establish stable HEK293 cell line.Methods: P2X7 gene was amplified by polymerase chain reaction from the human brain P2X7 cDNA and inserted into a vector pEGFP-N1 to construct a recombinant plasmidcalled pEGFP-N1/P2X7. The correct recombinant plasmid was transfected into HEK293 cells by X-fect transfection reagent. The cell line stably expressing EGFP tagged-P2X7 gene were established by screening with G418 and fluorescence microscope. The expression levels and localization of human P2X7 in HEK293 cells was identified by flow cytometry, Western blot and laser scanning confocal microscope. Results: The recombinant plasmid pEGFP-N1/P2X7 was constructed correctly and the stable HEK293 cell line expressing EGFP tagged-P2X7 fusion protein was established. Both Western blot and flow cytometry revealed the higher expression of humanP2X7 in the stably transfected HEK293 cells. Under the laser scanning confocal microscope the EGFP tagged-P2X7 fusion protein was located on the membrane of HEK293 cells.Conclusion: The eukaryotic expressing vector of pEGFP-N1/P2X7 is successfully constructed and the HEK293 cell line stably expressing P2X7-EGFP fusion protein is established which have provided solid experimental foundation for further studies on the structure and function of P2X7 ionic channel.
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    How to master the three important factors of research design in physiological study
    CHEN Fang, GUO Chun-xue, LIU Yi-song, SHEN Ning, HU Wan, YANG Meng-yuan, HU Liang-ping
    CJAP. 2016, 32 (5): 477-480.   DOI: 10.13459/j.cnki.cjap.2016.05.024
    Abstract   PDF (956KB) ( 13 )
    Objective: To offer a series of efficient methods for physiologists to make a scientific and rational experimental program. Methods: We bring about two questions as follows:Which are thecommon mistakesof statistics in the process of a physiological research? How to identify all the three essential factors? From the answers to these two questions, we highlight the methods to conduct a rigorous and accurate research design.Results: We summarized the special content of three essential factors in the experimental design:subject, influential factors, and effective indicators, which provide a basis for the design of the experimental program. Conclusion: In statistics studies, three essential factors which are "subject" "influential factors" "effective indicators" should be given considerable attention by the physiologists.
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