1.Role of GSK-3β in the protective effect of propofol pretreatment against hepatic ischemia-reperfusion injury and oxidative stress in rats
Yi CAO ; Na DUAN ; Yulin ZHU ; Xin SHEN ; Qiang WANG ; Ge ZHAO
Chinese Journal of Hepatobiliary Surgery 2016;22(10):692-696
Objective To investigate the protective effect of propofol pretreatment against hepatic ischemia-reperfusion injury and oxidative stress in rats and the mechanism of the role of GSK-3 β.Methods Sixty SD rats were randomly divided into four groups:sham operation group (S group),ischemia-reperfusion group (I-R group),propofol pretreatment group (P group),TDZD-8 pretreatment group (T group).The hepatic ischemia-reperfusion rat models were established by the method of Nauta.Rats were subjected to 30-min,60-min and 90-min 70% warm ischemia of liver followed by reperfusion for 120 min,respectively.Propofol (12 mg/kg · h) was injected via femoral vein 30 min before ischemia till the end of reperfusion in P group and TDZD-8 (1 mg/kg) were injected via femoral vein 20 min before ischemia in T group.The animals were killed at 120 min after reperfusion.Blood samples and the liver tissue were obtained.The levels of alanine aminotransferase (ALT),aspartate aminotransferase (AST),lactate dehydrogenase (LDH),malondialdehyde (MDA) and superoxide dismutase (SOD) were analyzed.Liver morphological changes were observed using optical microscopy.p-GSK-3β Ser9 and total GSK-3 β expression was determined by Western blot.Results Compared with S group,AST,ALT,LDH and MDA level was increased,SOD level was reduced,and p-GSK-3 β Ser9 expression was significantly reduced in I-R group.Compared with I-R group,the content of AST,ALT,LDH and MDA was reduced significantly,SOD increased significantly,and the content of p-GSK-3β Ser9 increased significantly in P group and T group.There were no significant differences between P group and T group.The hematoxylin-eosin staining of hepatic tissues revealed in I-R group had severe structural damage and periportal inflammatory cells infiltrated,hepatocyte necrosis and sinusoidal congestion.In P group and T group,liver tissues had normal structure,less cell death,edema and inflammatory cell infiltration.Conclusions Propofol can significantly reduce hepatic ischemia reperfusion injury by reducing oxidative stress and lipid hydroperoxides.This protective effect of Propofol may be associated with the inhibition of GSK-3 β by GSK-3 β Ser9 phosphorylation.
2.Primary culture and growth characteristics of four different species of lens epithelial cells
Li-Xia, JI ; Cai-Na, LI ; Quan, LIU ; Yi, HUAN ; Shuai-Nan, LIU ; Zhu-Fang, SHEN
International Eye Science 2015;(7):1151-1153
AlM:To explore the primary culture conditions for four kinds of lens epithelial cells ( LECs) of rat, rabbit, dog, and human, and measure their growth characteristics.METHODS:The lens capsule or anterior capsular tissue of rat, rabbit, dog and patient were removed by different methods, and they were cut into tiny pieces for primary culture by modified tissue adherent method. The morphological features of four kinds of LECs were observed under an inverted microscope.RESULTS: Four kinds of LECs of rat, rabbit, dog and human could be cultured primarily by tissue adherent method. With the evolution of tissue source, the adherent capacity of LECs gradually strengthened, cells form were changed from irregular polygon to oval, nucleus rounded and cytoplasm enriched gradually. Four kinds of LECs had fibrotic changes after several passages.CONCLUSlON: LECs of rat, rabbit, dog and human can be primarily cultured. This method lays the foundation for the mechanism research of caratact and related fields on the cellular and molecular levels.
3.Advances of the mechanism study on berberine in the control of blood glucose and lipid as well as metabolism disorders.
Ning SHEN ; Cai-Na LI ; Yi HUAN ; Zhu-Fang SHEN
Acta Pharmaceutica Sinica 2010;45(6):699-704
Berberine, an isoquinoline alkaloid isolated from some Chinese medicinal herbs such as Coptidis rhizoma, has been used for the treatment of diarrhea and other gastrointestinal infections as an antibacterial drug in Chinese medicine. In recent years, it was reported to have beneficial effects on the metabolism disorders states of diabetes. The mechanisms involve many aspects of the diabetes, including regulating the blood cholesterol and triglyceride, lowering blood glucose, ameliorating the insulin resistant state and influencing the function of the pancreatic beta cell.
Animals
;
Berberine
;
isolation & purification
;
pharmacology
;
Blood Glucose
;
metabolism
;
Coptis
;
chemistry
;
Diabetes Mellitus
;
metabolism
;
Drugs, Chinese Herbal
;
isolation & purification
;
pharmacology
;
Humans
;
Insulin
;
metabolism
;
secretion
;
Insulin-Secreting Cells
;
drug effects
;
Metabolic Diseases
;
metabolism
;
Nicotinamide Phosphoribosyltransferase
;
biosynthesis
;
genetics
;
Plants, Medicinal
;
chemistry
;
Protein Kinases
;
metabolism
;
RNA, Messenger
;
metabolism
;
Receptors, LDL
;
genetics
;
metabolism
;
Signal Transduction
4.Toll like receptor 9 expression and functions of plasmacytoid dendritic cells in the peripheral blood of patients with chronic hepatitis B virus infection
Ni-Na JIA ; Qing XIE ; Bao-Yan AN ; Lan-Yi LIN ; Huai-Cheng SHEN ; Hui WANG ; Xia-Qiu ZHOU ; Hong YU ; Qing GUO ;
Chinese Journal of Infectious Diseases 2007;0(12):-
Objective To investigate the expression of Toll-like receptor(TLR) 9 of circulating plasmacytoid dendritic cell (pDC) and analyze the frequency and interferon (IFN)-?production of circulating pDC during hepatitis B virus(HBV) infection.Methods Peripheral blood was collected from 69 HBV-infected patients,including 14 cases of asymptomatie HBV infection,30 cases of chro- nic hepatitis B(CHB),25 cases of HBV-related liver cirrhosis,and 21 healthy blood donors as con- trols.Flow cytometry was used to analyze the frequency of circulating pDC and the mean fluorescence intensity(MFI) of TLR9.Fresh peripheral blood mononuclear cells(PBMC) were stimulated by CpG ODN 2216 for 24 h in vitro.IFN-?in the supernatant was measured using enzyme-linked immunosorbent assay(ELISA) to analyze the frequency and IFN-?production of pDC during HBV in- fection.Data were analyzed with SPSS 13.0 for windows.Results Compared with healthy controls (62.6?10.7),the MFI of TLR9 of patients with asymptomatic HBV infection,those of CHB and HBV-related cirrhosis were significantly reduced (P
5.Oridonin inhibits proliferation of Jurkat cells via the down-regulation of Brg1.
Zhen-Zhen YE ; Fei-Long XUE ; Wen-Ping DING ; Xiang KONG ; Yi-Na SHEN
Chinese Journal of Contemporary Pediatrics 2017;19(11):1208-1212
OBJECTIVETo investigate the effect of oridonin on the human acute lymphocytic leukemia cell line Jurkat and its mechanism.
METHODSJurkat cells were cultured in vitro and treated with various concentrations (0, 1.25, 2.5, 5, and 10 μmol/L) of oridonin for different lengths of time (24, 48, and 72 hours). The proliferation of Jurkat cells was analyzed by MTT assay. The changes in nuclear morphology were evaluated by fluorescence microscopy at 12 hours after treatment with various concentrations of oridonin. The expression levels of Brg1, P53, and C-myc were determined by semi-quantitative Western blot in Jurkat cells treated with various concentrations of oridonin for 24 hours or 5 μmol/L oridonin for various lengths of time (0, 2, 6, 12, and 24 hours). The expression levels of P53 and C-myc and proliferation of Jurkat cells were evaluated after Brg1 expression was knocked down by Brg1-specific siRNA.
RESULTSCompared with the control group, the proliferation of oridonin-treated Jurkat cells was significantly inhibited in a concentration- and time-dependent manner (P<0.05). According to the florescence microscopic analysis, oridonin treatment led to nuclear pyknosis in Jurkat cells. Compared with the control group, Jurkat cells treated with 5 μmol/L oridonin had reduced expression of Brg1 and C-myc but elevated expression of P53. Brg1 knock-down led to a significant reduction in proliferation of Jurkat cells (P<0.05), up-regulated expression of P53, and down-regulated expression of C-myc.
CONCLUSIONSOridonin can inhibit the proliferation of Jurkat cells, probably via the Brg1 signaling pathway.
Antineoplastic Agents, Phytogenic ; pharmacology ; Cell Proliferation ; drug effects ; DNA Helicases ; analysis ; physiology ; Diterpenes, Kaurane ; pharmacology ; Dose-Response Relationship, Drug ; Down-Regulation ; Humans ; Jurkat Cells ; Nuclear Proteins ; analysis ; physiology ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; drug therapy ; Proto-Oncogene Proteins c-myc ; analysis ; Signal Transduction ; physiology ; Transcription Factors ; analysis ; physiology ; Tumor Suppressor Protein p53 ; analysis
6.Inhibition effect of epalrestat on rat lens osmotic expansion.
Li-Xia JI ; Ning SHEN ; Cai-Na LI ; Quan LIU ; Yi HUAN ; Zhu-Fang SHEN
Acta Pharmaceutica Sinica 2009;44(10):1107-1111
Epalrestat is the unique aldose reductase inhibitor on the market, which was mainly used for the diabetic neuropathy. Lenses osmotic expansion could be induced by galactose to mimic the pathological process of diabetic cataract in vitro. In present study, we mainly investigated whether epalrestat possesses inhibitory effect on the lens osmotic expansion. The results indicated that epalrestat could not only markedly inhibit rat lens osmotic expansion in vitro, but also significantly reduced the high expression of the osmotic expansion-related genes such as AR and AQP1 in mRNA and protein levels. The findings may provide an important reference to epalrestat in the clinical application for the treatment of diabetic cataract.
Aldehyde Reductase
;
antagonists & inhibitors
;
biosynthesis
;
genetics
;
Animals
;
Aquaporin 1
;
genetics
;
metabolism
;
Cataract
;
etiology
;
metabolism
;
pathology
;
Diabetes Mellitus, Experimental
;
complications
;
Enzyme Inhibitors
;
pharmacology
;
Galactose
;
antagonists & inhibitors
;
Lens, Crystalline
;
drug effects
;
metabolism
;
pathology
;
Male
;
Osmosis
;
drug effects
;
RNA, Messenger
;
metabolism
;
Rats
;
Rats, Sprague-Dawley
;
Rhodanine
;
analogs & derivatives
;
pharmacology
;
Thiazolidines
;
pharmacology
7.Expression of A-kinase anchor protein 95, cyclin E2, and connexin 43 in lung cancer tissue, clinical significance of their expression, and their expression correlation.
Yi-de CHEN ; Xiao-xuan CHEN ; Li-na SHEN ; Feng-chao LIANG ; Ye DING ; Xiu-yi YU ; Mao-qiang XUE ; Yong-xing ZHANG
Chinese Journal of Industrial Hygiene and Occupational Diseases 2012;30(10):725-729
OBJECTIVETo study the expression of A-kinase anchor protein 95 (AKAP95), cyclin E(2), and connexin 43 (Cx43) in lung cancer tissue, the clinical significance of their expression, and the expression correlation among the three proteins.
METHODSFifty-one samples of lung cancer tissue were examined by immunohistochemistry to measure the expression of AKAP95, cyclin E2, and Cx43.
RESULTSThe positive rate of AKAP95 expression in lung cancer tissue was significantly higher than that in paracancerous tissue (82.35% vs 33.33%, P < 0.05); AKAP95 expression was associated with the cell differentiation and histopathological type of lung cancer (P < 0.05). The positive rate of cyclin E(2) expression in lung cancer tissue was significantly higher than that in paracancerous tissue (43.14% vs 13.33%, P < 0.05); cyclin E(2) expression was associated with the lymph node metastasis and histopathological type of lung cancer (P < 0.05). The positive rate of Cx43 expression in lung cancer tissue was lower than that in paracancerous tissue (60.78% vs 80.00%); Cx43 expression was associated with the cell differentiation, lymph node metastasis, and histopathological type of lung cancer (P < 0.05). There was correlation between each two of AKAP95 expression, cyclin E(2) expression, and Cx43 expression in lung cancer tissue.
CONCLUSIONHigh expression of AKAP95 and cyclin E(2) plays an important role in the occurrence and development of lung cancer. AKAP95 expression is associated with the cell differentiation and histopathological type of lung cancer, and cyclin E2 expression is associated with lymph node metastasis and histopathological type. There is correlation between each two of AKAP95 expression, cyclin E(2) expression, and Cx43 expression in lung cancer tissue.
A Kinase Anchor Proteins ; metabolism ; Adult ; Aged ; Connexin 43 ; metabolism ; Cyclins ; metabolism ; Female ; Humans ; Lung ; metabolism ; pathology ; Lung Neoplasms ; metabolism ; pathology ; Lymphatic Metastasis ; Male ; Middle Aged
8.Influence of chronic diseases on health related quality of life in middle-aged and elderly people from rural communities: application of EQ-5D scale on a Health Survey in Fangshan, Beijing
Yang CAO ; Xun TANG ; Li YANG ; Na LI ; Yi-Qun WU ; Wen-Yi FAN ; Jian-Jiang LIU ; Li-Ping YU ; Hai-Tao XU ; Wu-Jun LIU ; Su-Shen WEN ; Yong-Hua HU
Chinese Journal of Epidemiology 2012;33(1):17-22
Objective To evaluate the health related quality of life (HRQoL) and to explore the influencing factors related to the prevalence of the essential hypertension,diabetes,coronary heart disease (CHD) and stroke in middle-aged and elderly people from rural communities.Methods A total of 5345 individuals were chosen from a 2010 Health Survey in Fangshan,Beijing.Self-designed questionnaire was applied to collect the information on general health-related behaviors,chronic conditions,self-rated health,blood pressure and HRQoL with the EuroQoL questionnaire (EQ-5D) were measured.Probit regression model was used to identify the influencing factors on HRQoL,adjusting age,marriage status,education level,smoking,alcohol intake,physical activity and control of blood pressure.Population Attributable Risk Proportion (PARP) for estimating the influence of chronic disease on HRQoL was also calculated.Results The average EQ-5D index was 0.923 ±0.145,with 72.1% of the overall respondents in the status of having chronic diseases.Participants with chronic diseases had significantly lower EQ-5D index (0.907 ± 0.155 ) than those without (0.963 ±0.105 ) (t =15.082,P< 0.001 ).Significant relations were found between a lower HRQoL and chronic diseases.Relative risk (RR) and 95% confidence interval (95% CI) of poor HRQoL in populations with hypertension,diabetes,CHD and stroke were 1.686 (95%CI: 1.519-1.871),1.725 (95%CI:1.502-1.981 ),1.869 (95%CI: 1.663-2.099),and 1.759 (95%CI: 1.474-2.089) respectively.The most influential disease appeared to be stroke in male (RR=1.929,95%CI: 1.462-2.546),whereas CHD in female individuals (RR=1.926,95%CI: 1.671-2.220).Rates of PARP (PAR%) for poor HRQoL in chronic disease subgroups were as follow: hypertension 31.05%,CHD 23.39%,diabetes 4.84% and stroke 4.44%,respectively.Conclusion This study suggested that stroke,at individual level,was the major influential factor to HRQoL.However,hypertension remained the leading preventable influential factor to HRQoL at the population level.
9.Expressions and activities of Rho GTPases in hypoxia and its relationship with tumor angiogenesis.
Yan XUE ; Feng BI ; Wen-chao LIU ; Yang-lin PAN ; Zhe-yi HAN ; Na LIU ; Chang-jiang LIU ; Hui-qin SHEN ; Xue-yong ZHANG
Chinese Journal of Oncology 2004;26(9):517-520
OBJECTIVETo study the expressions and activities of Rho GTPases in hypoxia and its relationship with tumor angiogenesis.
METHODSThree tumor cell lines were used in this study: gastric cancer cell lines AGS, SGC7901 and hepatocellular carcinoma cell line HepG2. Expression level of Rac1 mRNA was detected by semi-quantitative RT-PCR. Activity of Rac1 was determined by pull-down assay and expression of HIF-1alpha, VEGF, p53 and PTEN protein was detected by Westernblot.
RESULTSThe expression level of Rac1 mRNA was significantly increased in hypoxia compared to normoxia. Pull-down assay showed that hypoxia-induced activity of Rac1 was elevated in a time-dependent manner and climaxed at 3 hours. The expressions of HIF-1alpha and VEGF protein were up-regulated, while those of PTEN and p53 protein were down-regulated.
CONCLUSIONThese results indicate that hypoxia enhances Rac1 expression which might be involved in tumor angiogenesis by reacting with hypoxia-responsive genes.
Cell Hypoxia ; Cell Line, Tumor ; Hepatoblastoma ; blood supply ; metabolism ; pathology ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; biosynthesis ; genetics ; Liver Neoplasms ; blood supply ; metabolism ; pathology ; Neovascularization, Pathologic ; PTEN Phosphohydrolase ; biosynthesis ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Stomach Neoplasms ; blood supply ; metabolism ; pathology ; Tumor Suppressor Protein p53 ; biosynthesis ; genetics ; Vascular Endothelial Growth Factor A ; biosynthesis ; genetics ; rac1 GTP-Binding Protein ; biosynthesis ; genetics ; rho GTP-Binding Proteins ; biosynthesis ; genetics
10.Expression and prognostic value of M2 macrophages and regulatory T cells in gastric carcinoma tissue.
Ping SHU ; Jing QIN ; Xin-Yu QIN ; Yi-Hong SUN ; Zhen-Bin SHEN ; Yan-Na WANG ; Jing-Lei LIU
Chinese Journal of Gastrointestinal Surgery 2011;14(5):368-371
OBJECTIVETo explore the prognostic value of M2 macrophages and regulatory T cells(Tregs) in gastric carcinoma.
METHODSClinicopathological characteristics and follow up data of 135 patients with gastric carcinoma were collected. Patients included were those who underwent D2 radical resection(R0) at Zhongshan Hospital of Fudan University from February 1999 to December 2005. Tissue chips of gastric carcinoma specimen were stained using immunohistochemistry to determine the cells density and number of M2(CD163 positive) and Tregs(Foxp3 positive).
RESULTSThe median positive cells density of M2 macrophages and Tregs in tumor tissue were 7.48/HP and 6.33/HP, respectively, higher than that in adjacent tissues(1.37/HP and 2.92/HP, P<0.001). The density of M2 macrophages was positively correlated with that of Treg cells(r=0.415, P<0.001) in tumor tissue. The median survival of patients with low expression of M2 and Tregs(n=43) was significantly longer than those with high expression of the 2 cells(n=45) (99.0 vs. 72.3 months, P<0.05).
CONCLUSIONCombined detection of M2 macrophages and Tregs may predict the prognosis of gastric carcinoma.
Adult ; Aged ; Aged, 80 and over ; Antigens, CD ; metabolism ; Antigens, Differentiation, Myelomonocytic ; metabolism ; Female ; Forkhead Transcription Factors ; metabolism ; Humans ; Macrophages ; metabolism ; pathology ; Male ; Middle Aged ; Prognosis ; Receptors, Cell Surface ; metabolism ; Stomach Neoplasms ; diagnosis ; metabolism ; pathology ; T-Lymphocytes, Regulatory ; metabolism ; pathology ; Young Adult