1.Expression of hepatic signal transduction proteins following reduced-size liver transplantation in rats
Jing LIU ; Li LI ; Jianghua RAN ; Shengning ZHANG ; Laibang LI ; Yang GAO ; Yiming CHEN ; Xibing ZHANG
Chinese Journal of Tissue Engineering Research 2014;(49):7974-7978
BACKGROUND:The proteome is a highlight technology in medical research fields lately, and has been reported to be applied in basic research fields related to liver transplantation. However, it has not been heard that the proteome has been used in research related to reduced-size liver transplantation. OBJECTIVE: To study expression of hepatic differential proteins related to signal transduction using proteomics after reduced-size liver transplantation in rats. METHODS:On the basis of successful establishment of rat models of reduced-size liver transplantation, transplanted liver tissues were obtained at 1, 3 and 7 days after transplantation. Postoperative liver tissue and normal donor, receptor liver tissues were subjected to solid pH gradient two-dimensional gel electrophoresis. Two-dimensional gel electrophoresis patterns were set up. Differentialy expressed protein spots were identified using tandem mass spectrometry analysis and database. RESULTS AND CONCLUSION:Seventy-two differential protein stains were found taking 10 times measure. Finaly, 32 proteins with clear functions were identified. Of them, four proteins participated in signal transduction, and they distributed at 3 and 7 days after liver transplantation, accounting for 6%. Results verified that on the basis of successful and stable establishment of rat models of reduced-size liver transplantation, proteomics technology was utilized to study differential proteins involving in signal transduction after reduced-size liver transplantation, and this study provides data for further deep investigation of regulating MicroRNA of these proteins.
2.Plasma free fatty acids composition and its association with nonalcoholic fatty liver disease
Li RAN ; Jing WAN ; Haiying ZHANG ; Shihui CHEN ; Yanxiang GAO ; Qianyong ZHANG ; Mantian MI
Chongqing Medicine 2014;(14):1697-1699
Objective To study the relationship between plasma free fatty acids composition and the incidence of nonalcoholic fatty liver disease(NAFLD) .Methods By the design of case‐control study ,105 patients with NAFLD as cases and 110 healthy peo‐ple as controls were enrolled into the study .Plasma free fatty acid levels were determined by gas chromatography .Results High level of plasma palmitic acid(C16 :0)(OR=1 .769) was the risk factors of NAFLD ,while plasma levels of linoleic acid(C18 :2 n‐6) (OR=0 .855) and arachidonic acid(C20 :4 n‐6)(OR=0 .181)were negatively associated with the incidence of NAFLD .Conclusion These findings suggest that a proper ratio of diet fatty acids intake may reduce the risk of NAFLD .
3.Literature Research of Mild Cognitive Impairment Syndrome Distribution Characteristics
Yintong GAO ; Jinzhou TIAN ; Jing SHI ; Ran ZHANG ; Huishang FENG ; Kaihang GUO ; Xiaoyuan JIANG
Journal of Traditional Chinese Medicine 2017;58(17):1503-1506
Objective To explore the Chinese medicine syndrome characteristics of mild cognitive impairment (MCI).Methods The papers of MCI syndrome research were reviewed and collected in China National Knowledge Infrastructure,Wan Fang Data and VIP Data from January of 1990 to December of 2014.Statistical analysis was made on the Chinese medicine syndrome types and syndrome factors.SPSS 17.0 software was adopted to make cluster analysis.Combined with experts' experience,related symptoms to the syndrome factors were carried out.Results Totally 32 papers were included.Mter terminology normalization,there were 24 syndrome types of MCI.Top 5 syndromes with high frequency were syndrome of orifices confused by phlegm,syndrome of deficiency of kidney essence,syndrome of deficiency of both Qi and blood,syndrome of internal exuberance of heat toxin and syndrome of blood stasis blocking brain.In syndrome factors of disease location type,kidney and brain covered the highest proportion,30.83% and 30.00%.In syndrome factors of disease cause and character types,Qi deficiency covered the most,16.50%.According to results of cluster analysis,combined with experts' experience,15 syndrome factors were extracted,including 69 symptoms.Conclusion Chinese medicine syndrome types of MCI were mainly syndrome of orifices confused by phlegm and syndrome of deficiency of kidney essence.The disease locations were mainly kidney and brain.The disease character was Qi deficiency.
4.Establishment of two human tumor cell lines with lentivirus-mediated stably Smurf1 silencing and their effect on cell migration
Rongfei WEI ; Jing GUO ; Mengyuan LI ; Ruimin ZHU ; Xingjiu YANG ; Ran GAO
Chinese Journal of Comparative Medicine 2017;27(4):46-51
Objective To establish lentiviral expression vectors for Smurf1 silencing and assess the effects of Smurf1 silencing on cell migration.Methods HeLa and A549 cells were infected with lentiviral expression vectors for Smurf1 silencing respectively.After 7 days,the stable cell lines with Smurf1 silencing were obtained after puromycin-resistance screening,enrichment and expansion.The intracellular gene and protein levels of Smurf1 were detected by qPCR and western blot.Transwell assay was used to assess the effect of Smurf1 silencing on cell migration.Results The stable cell lines with Smurf1 silencing are constructed successfully.Silencing of Smurf1 down-regulated cell migration rate detected by Transwell assay.Conclusion Smurf1 promotes cell migration.
5.Expression of hepatic energy proteins following reduced-size liver transplantation in rats
Jing LIU ; Li LI ; Jianghua RAN ; Shengning ZHANG ; Laibang LI ; Xibing ZHANG ; Yang GAO ; Yiming CHEN
Chinese Journal of Tissue Engineering Research 2015;(18):2874-2878
BACKGROUND:At present, the proteome is a mature technology that has been applied in basic research fields related to liver transplantation. But, it has been not reported in research related to reduced-size liver transplantation.
OBJECTIVE:To explore the expression of differential proteins related to hepatic energy metabolism fol owing reduce-size liver transplantation in rats by using by proteomic technology.
METHODS:The improved model of reduced-size liver transplantation was used in this experiment. The donor was health female Lewis rats and the recipient was male Wistar rats for liver transplantation. The difference between the donor and the recipient was about 20 g. The weight of donor liver/the weight of recipient donor was approximately equal to 50%. The donor liver tissue was harvested and trimmed to the required size. The portal vein and infrahepatic vena cava were cannulated, and the biliary tract was implanted into the donor bile duct for transplantation. Then the donor was transplanted into the recipient after the removal of original liver tissue. Hepatic specimens were harvested by 1, 3 and 7 days after reduced-size liver transplantation. Then, the harvested specimens were compared with the normal donor and recipient liver tissue that were previously harvested and frozen, to generate two-dimensional gel electrophoresis profile using proteome technology. Then tandem mass spectrometry and databases analysis were performed after two-dimensional electrophoresis for identifying differential protein stains.
RESULTS AND CONCLUSION:In this experiment, 72 differential protein stains with over lo-fold changes were selected. After identification, 32 proteins showed clear functions, and among them three differential proteins (ATP synthase beta subunit, electron-transferring flavoprotein beta peptide and proton-transferring ATP synthase) were involved in the process of cel energy metabolism. The proteins were distributed on 1 and 7 days after reduce-size liver transplantation, accounting for 6%.
6.A Model of Focal Cortical Infarctionin Rat:Mini mally Invasive Craniotomy
Jing XUE ; Pei-yi GAO ; Yi-hua AN ; Chong-ran SUN ; Jin LI ; Hua HUANG
Chinese Journal of Rehabilitation Theory and Practice 2006;12(1):11-13
ObjectiveTo develop a stable model of focal cerebral infarction in rat to study the curative effect of neural stem cells transplantation.MethodsThirty-seven rats were selected which were divided into two groups in random, experimental group and control group. The focal infarction model was developed by the ligation of the left middle cerebral artery followed by the ligation of the ipsilateral common carotid artery and the temporary clip occlusion of the contralateral common carotid artery for 1.5 h. The operation adopted minimally invasive craniotomy though temporal bone. The model was evaluated by examining the neurologic deficits, ink perfusion, TTC staining and Magnetic Resonance imaging.ResultsAll the rats were in good condition after the operation, the mortality rate was 6.25% after 4 weeks. Ink perfusion and TTC staining confirmed that the ischemia was confined to the cortex. The areas of infarction measured 83.52 mm3 by Magnetic Resonance imaging after 4 weeks.ConclusionA stable focal cerebral infarction model can be achieved by minimally invasive craniotomy. It is superior for its homogeneity of infarction volume and site, and its low mortality. It can be used for the study of transplantation of neural stem cells.
7.Protective effect of DIZE on heart function of rats with diabetic cardio-myopathy
Min YANG ; Xin-Ran CAO ; Yuan-Yuan WANG ; Xiao-Qiong WANG ; Shi-Ran YU ; Bo DONG ; Jing GAO
Chinese Journal of Pathophysiology 2018;34(1):147-151,177
AIM:To observed the protective effect of diminazene aceturate(DIZE),an angiotensin-converting enzyme 2(ACE2)activator,on rats with diabetic cardiomyopathy(DCM).METHODS:Male Wistar rats(n=30)were randomly divided into normal control group ,DCM group and DIZE treatment group(DIZE group).The rats in DCM group and DIZE group were intraperitoneally injected with streptozotocin(65 mg/kg )to establish diabetic model.After 12 weeks,the diabetic rats were infused with DIZE at 15 mg· kg-1 · d-1 or the same volume of saline for 4 weeks using os-motic minipump.The cardiac function was measured at the end of the 16th week.The methods of Mason staining and HE staining were used to observe the morphological changes of the myocardial tissue.Western blot ,ELISA and immunohisto-chemistry were used to observe the changes of ACE2,angiotensin(Ang)Ⅱ,Ang-(1-7),interleukin(IL)-1,IL-6 and connective tissue growth factor(CTGF).RESULTS:DIZE significantly improved the expression of ACE 2 in diabetic rats(P<0.05).Compared with DCM group,the levels of IL-1 and IL-6 in DIZE group were significantly decreased ,and the cardiac function in DIZE group was significantly improved(P<0.05).CONCLUSION:ACE2 endogenous agonist DIZE significantly increases the ACE 2 level and reduces the level of inflammation ,thus protecting the heart function of DCM rats.
9.Interleukin-6 expression in rhesus monkey models with acute rejection after liver transplantation
Jianghua RAN ; Xibing ZHANG ; Jing LIU ; Shengning ZHANG ; Laibang LI ; Yang GAO ; Yiming CHEN ; Wang LI ; Li LI
Chinese Journal of Tissue Engineering Research 2014;(36):5758-5763
BACKGROUND:Interleukin-6 is an important cytokine in the immune inflammatory response, strongly links with graft rejection reaction, and plays an important role in diagnosis of graft rejection and evaluation of anti-rejection. OBJECTIVE:To measure the expression of interleukin-6 in acute rejection of the liver transplantation in the rhesus monkey, and to evaluate the value as an early diagnosis of acute rejection after liver transplantation. METHODS:A total of 16 rhesus monkeys were used as the object and randomly divided into experimental group (no treated by immunosuppressant in perioperative period), and control group (treated by immunosuppressant in perioperative period). The al ograft orthotopic liver transplantation models were established in those monkeys. Then serum and liver tissue were col ected at 6, 12, 24, and 72 hours after surgery. Al ograft rejection was monitored by liver function tests, and hematoxylin-eosin staining of liver and Banff score. Final y, the expression levels of interleukin-6 were detected by enzyme linked immunosorbent assay and immunohistochemistry.RESULTS AND CONCLUSION:Acute graft rejection reaction appeared at 12, 24 and 72 hours after liver transplantation in the experimental group. The expressions of alanine aminotransferase, aspartate aminotransferase and total bilirubin were significantly higher in the experimental group than in the control group at 24 and 72 hours (P<0.05). Histological manifestations were severer and Banff score was higher in the experimental group at 72 hours than in the control group (P<0.05). Interleukin-6 levels were significantly higher in the serum and liver tissue of experimental group than in the control group at 12, 24 and 72 hours after liver transplantation (P<0.05), especial y at 72 hours. Results suggested that interleukin-6 possibly participated in rejection after liver transplantation. The expression of interleukin-6 was probably of significance in the early diagnosis of acute rejection after orthotopic liver transplantation in rhesus monkeys.
10.Variation of tumor necrosis factor-alpha during acute rejection after liver transplantation in a rhesus monkey
Xibing ZHANG ; Jianghua RAN ; Jing LIU ; Shengning ZHANG ; Laibang LI ; Yiming CHEN ; Yang GAO ; Wang LI ; Li LI
Chinese Journal of Tissue Engineering Research 2014;(36):5752-5757
BACKGROUND:Tumor necrosis factor-αis an inflammatory cytokine involved in the immune response and increasing graft antigen expression. OBJECTIVE:To investigate the relationship between tumor necrosis factor-αin the liver tissue and acute rejection after liver transplantation in a rhesus monkey. METHODS:Liver transplant models in rhesus monkey were constructed by the improved vascular dual cuff, supporting tube of biliary tract and artery anastomosis method. The successful models were randomly divided into experimental group (no immunosuppressant treatment in perioperative period) and control group (treated by immunosuppressant in perioperative period). Then the blood samples and liver tissue were col ected at 6, 12, 24, and 72 hours after surgery. Al ograft rejections of liver transplantation were monitored by liver function tests, hematoxylin-eosin staining and Banff score. Final y, the expression level of tumor necrosis factor-αwas detected by western blot analysis and immunohistochemistry technique. RESULTS AND CONCLUSION:The expression of tumor necrosis factor-αin the experimental group and control group began to increase at 6 hours, reached the peak at 12 hours, and then decreased at 24-72 hours. The changes of expression level were the most obvious in the experimental group. At 6, 12, 24 and 72 hours, the expression of tumor necrosis factor-αin the experimental group was significantly higher than that in the control group (P<0.05). This change appeared earlier than pathological changes in the liver and liver function. Variations in the expression of tumor necrosis factor-αafter liver transplantation have important implications for early diagnosis of acute rejection after liver transplantation.