2.Experimental study on apoptosis induced by pcDNA3-survivin-mutant in gastric cancer cell lines
Jihong TAN ; Shuiping TU ; Bing ZOU ; Tianle MA ; Jie ZHONG ; Chenli ZHANG ; Minmin QIAO ; Shihu JIANG ;
Chinese Journal of Digestion 2001;0(04):-
Objective Survivin is overexpressed in gastric cancer. However it not expressed in normal gastric mucosa. The expression of survivin is tightly related to the prognosis of gastric cancer.By gene reconstruction we generated pcDNA3 survivin mutant(Cys84Ala) plasmid, and observed its effect on the gastric carcinoma cell lines. Methods The survivin mRNA and protein expression levels were determined by reverse transcription polymerase chain reaction(RT PCR) analysis,Western blot and immunohistochemical staining respectively . Flowcytometry and acridine orange staning were employed to detect apoptosis. Results Overexpression of survivin mRNA and protein were detected in the gastric cancer cell lines. Inhibition of survivin by survivin mutant cDNA induced apoptosis,activated caspase 3 activity,cleaved PARP and promoted cytochrome C releasing in gastric cancer cells,and effectively sensitized gastric cancer cells to chemotherapeutic agents. Conclusion Inhibition of survivin may induce apoptosis in gastic cancer and sensitize gastric cancer cells to chemotherapeutic agents.Survivin targeted therapeutic protocol may potentially benefit gastric cancer therapy.
3.Effect of compound qizhu granule on cellular immunity of chronic hepatitis B patients.
Yin-fang ZHU ; Xi-bing GU ; Xiao-ye GUO ; Zhi-han YAN ; Yun-chuan PU ; Kang-wan TU ; Zhong HUA ; Hao PEI
Chinese Journal of Integrated Traditional and Western Medicine 2014;34(10):1178-1181
OBJECTIVETo explore the effect of compound qizhu granule (CQG) on cellular immunity of chronic hepatitis B (CHB) patients.
METHODSTotally 103 CHB patients treated with lamivudin (LAM) for 6 months, who had partial virological response (HBeAg positive) were randomly assigned to two groups, 50 in the treatment group and 53 in the control group. All patients took LAM 100 mg (once a day) plus ADV 10 mg (once a day). Patients in the treatment group additionally took CQG, one dose per day. After one-year treatment hepatitis B virus (HBV) DNA negative rates, HBeAg seroconversion, levels of HBV specific cytotoxic T lymphocyte (CTL), non-specific CTL and natural killing (NK) cells were compared between the two groups.
RESULTSAfter 1-year treatment, HBV DNA negative rate of the treatment group was 88: 0% in 44 cases, slightly higher than that of the control group (41 cases, 77.4%), but with no statistical difference (P >0.05). HBeAg seroconversion of the treatment group was 32.0% in 16 cases, higher than that of the control group (8 cases, 15.1%), with statistical difference (P <0.05). Levels of HBV specific CTL (0.79%±0. 07%), non-specific CTL (19.4%±1.8%) and NK cells (14. 1%± 1.5%) of the treatment group were higher than those of the control group (0.58% ± 0.08%, 17.5% ± 1.7%, and 11.1%±1.5%, respectively; allP <0.01).
CONCLUSIONTreating CHB patients with partial virological response by ADV plus CQG could improve specific and non-specific cellular immunity, thereby elevating HBeAg seroconversion rate.
Drugs, Chinese Herbal ; therapeutic use ; Hepatitis B e Antigens ; immunology ; Hepatitis B virus ; genetics ; Hepatitis B, Chronic ; drug therapy ; immunology ; Humans ; Immunity, Cellular ; immunology ; T-Lymphocytes, Cytotoxic ; drug effects
4.Isolation and structure identification of chemical constituents from Saposhnikovia divaricata (Turcz.) Schischk.
Yan-Yan JIANG ; Bin LIU ; Ren-Bing SHI ; Guang-Zhong TU
Acta Pharmaceutica Sinica 2007;42(5):505-510
Fourteen compounds were isolated from the ethanol extraction of Saposhnikovia divaricata (Turcz.) Schischk using column chromatographic methods after enrichment by macroporous adsorptive resins. They were identified as fangfengalpyrimidine (1), clemiscosin A (2), 5-hydroxy-8-methoxypsoralen (3), sec-O-glucosylhamaudol (4), hamaudol (5), nodakenetin (6), prim-O-glucosylcimifugin (7), cimifugin (8), 4'-O-beta-D-glucosyl-5-O-methylvisamminol (9), 5-O-methylvisamminol (10), marmesin (11), adenosine (12), daucosterol (13) and beta-sitosterol (14) by physico-chemical properties and spectral data. Compound 1 is a new compound. Compounds 2 and 3 were isolated from umbelliferae plants and Saposhnikovia divaricata (Turcz.) Schischk for the first time respectively.
Apiaceae
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chemistry
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Chromatography, Thin Layer
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Chromones
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chemistry
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isolation & purification
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Coumarins
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chemistry
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isolation & purification
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Heterocyclic Compounds, 4 or More Rings
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chemistry
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isolation & purification
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Magnetic Resonance Spectroscopy
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Mass Spectrometry
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Methoxsalen
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analogs & derivatives
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chemistry
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isolation & purification
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Molecular Conformation
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Molecular Structure
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Monosaccharides
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chemistry
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isolation & purification
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Plant Roots
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chemistry
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Plants, Medicinal
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chemistry
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Pyrimidines
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chemistry
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isolation & purification
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Resins, Synthetic
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Xanthenes
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chemistry
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isolation & purification
5.The association of hepatitis B virus genotype and the basal core promoter mutation in Qidong, China.
Yan JIN ; Yu ZHU ; Tao-yang CHEN ; Xia GUO ; Jing ZHANG ; Jin-bing WANG ; Wei-zhong CHEN ; Geng-sun QIAN ; Hong TU
Chinese Journal of Hepatology 2010;18(7):511-515
OBJECTIVESTo investigate the prevalence of hepatitis B virus (HBV) genotypes and the association with hepatocellular carcinoma (HCC) or basal core promoter (BCP) mutation in Qidong, China.
METHODSThe whole genome of HBV or X gene sequences were obtained from serum samples of HBV infected patients by using PCR and direct sequencing methods. Phylogenetic tree was constructed to determine the genotypes or subgenotypes of HBV.
RESULTSAccording to the phylogenetic tree constructed from full-length sequence of HBV, genotype C2 was predominant in Qidong area. It was prevalent in 44 out of the 48 cases (91.7%), whereas genotype B2 only existed in 4 cases (8.3%). No other genotypes or recombinant types were found in Qidong patients. The result of genotyping based on X gene sequence confirmed the above observation. In a total of 182 samples, 169 (92.9%) showed genotype C2 and 10 (5.5%) showed genotype B2. There were 3 (1.6%) patients showed a coinfection with C2 and B2. The infection rate of genotype C in Qidong was significantly higher than that in neighboring city Shanghai (chi(2) = 12.252, P less than 0.01). There was no significant difference of genotype distribution between HCC and chronic hepatitis groups (P is more than 0.05). The frequency of T1762/A1764 double mutation in genotype C2 (70.3%) was significantly higher than that in genotype B2 (30.8%, P less than 0.05). The other two types of point mutation which also occurred in BCP, i.e. T1766 and A1768, were only seen in genotype C2.
CONCLUSION(1) Genotype C2 is the predominant genotype in Qidong, China. (2) There is no association between genotype C and HCC in Qidong. (3) Genotype C has a higher prevalence of BCP mutation than genotype B.
Adolescent ; Adult ; Aged ; Carcinoma, Hepatocellular ; epidemiology ; virology ; Child ; Child, Preschool ; China ; epidemiology ; DNA, Viral ; genetics ; Female ; Genotype ; Hepatitis B virus ; genetics ; Hepatitis B, Chronic ; virology ; Humans ; Liver Neoplasms ; epidemiology ; virology ; Male ; Middle Aged ; Mutation ; Prevalence ; Viral Core Proteins ; genetics ; Young Adult
6.A novel triterpenoid saponin from Prunella vulgaris.
Lan-Zhen ZHANG ; Ya-Jian GUO ; Guang-Zhong TU ; Ren-Bing SHI ; Wen QIN ; Yin-Ying BA ; Fan YANG
Acta Pharmaceutica Sinica 2008;43(2):169-172
To study the constituents of the Prunella vulgaris L, the constituents were isolated by various column chromatography and the structures were identified on the basis of chemical and spectral analysis. One saponin compound (I) and one flavone glycoside compound (II) were obtained from Prunella vulgaris L. Their structures were elucidated as 16-oxo-17-demethyl-3beta,24-dihydroxylolean-12-en-3-O-beta-D-glucuronoside (I), and acacetin-7-O-beta-D-glucopyranoside (II). Compound I is a novel triterpenoid saponin and named as prunelloside A. Compound II was obtained for the first time from the Prunella genus.
Flavonoids
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isolation & purification
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Glucosides
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isolation & purification
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Molecular Structure
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Plants, Medicinal
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chemistry
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Prunella
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chemistry
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Saponins
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chemistry
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isolation & purification
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Triterpenes
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chemistry
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isolation & purification
7.Preventive and therapeutic mechanism of the herbal pair, Scutellariae Radix and Coptidis Rhizoma on dementia-like mice induced by D-galactose
Chang-Hua ZHANG ; Tong-Tong LIU ; Ke-Zhong DENG ; Ying-Chong CHEN ; Gui-Bing LIN ; He-Ping YE ; Xiu-Ying TU ; Jun-Qing SHENG
Chinese Traditional Patent Medicine 2018;40(3):524-529
AIM To investigate the preventive effects of herbal pair,Scutellariae Radix and Coptidis Rhizoma (SC),on Alzheimer's disease (AD),and its mechanism of action.METHODS Dementia mice induced by 8-week s.i.d subcutaneous injection of D-galactose (100 mg/kg),were simultaneously given respective,intragastric administration of SC crude drug at doses of 5,10,20 g/kg,or piracetam support at 0.75 g/kg,and isometrical distilled water was applied to the mice of normal control group.The mice had their learning and memory abilities checked by Morris water maze at intervals of four weeks and eight weeks since the start of the trial,and their blood and brain tissue biochemical indices measured at the end of the test.RESULTS Significantly shortened latent period in place navigation test and the time of enter into the original platform in the space exploration test were observed in the mice treated with 4-week D-galactose and SC (P <0.05 或 P <0.01).The 8-week intervention demonstrated SC capacity in the significant promotion of T-SOD activity,decreased blood MDA levels (P < 0.01)and the brain AchE levels,and increased brain GSH-Px activity (P < 0.01).CONCLUSION SC increases the concentration of acetylcholine in brain tissue and protects the central nervous tissue under oxidative stress,highlighting its therapeutic effect on AD.
8.Modulatory effect of aquaporin 5 on estrogen-induced epithelial-mesenchymal transition in prostate epithelial cells
Yu FAN ; Tu-Run SONG ; Qiang WEI ; Lu YANG ; Tao LIN ; Xiao-Bing FENG ; Xian-Ding WANG ; Zhong-Li HUANG ; Shi-Jian FENG
Chinese Medical Journal 2021;134(4):448-455
Background::Estrogen is involved in the pathophysiological process of benign prostatic hyperplasia (BPH), in which epithelial-mesenchymal transition (EMT) plays an important role. Upregulation of aquaporin (AQP) 5, which is directly activated by estrogen, has been reported to promote EMT in multiple cells. This study aimed to examine the effects of AQP5 on estrogen-induced EMT in the prostate.Methods::Normal prostate (NP) tissue samples without any histopathological changes and BPH tissue samples with pathologically confirmed hyperplasia were obtained. An EMT cell model was subsequently established by adding estradiol (E2) to RWPE-1 cells, after which AQP5 knockdown was performed. Tissue morphological and immunohistochemical features were examined using hematoxylin-eosin and immunohistochemical staining. Western blot analysis was performed to determine the expression of AQPs, estrogen receptors, and EMT-related proteins. Cell proliferation was assessed and supernatants were collected for enzyme-linked immunosorbent assay to determine transforming growth factor-β1 (TGF-β1) concentrations. Immunofluorescence staining was performed to assess protein expressions in RWPE-1 cells. Results::BPH tissues exhibited greater EMT (TGF-β1: 1.362 ± 0.196 vs. 0.107 ± 0.067, P = 0.003; vimentin: 1.581 ± 0.508 vs. 0.221 ± 0.047, P < 0.001; E-cadherin: 0.197 ± 0.188 vs. 1.344 ± 0.088, P < 0.001), higher AQP5 (1.268 ± 0.136 vs. 0.227 ± 0.055, P < 0.001) and estrogen receptor (ER) α (1.250 ± 0.117 vs. 0.329 ± 0.134, P < 0.001) expression but lower ERβ (0.271 ± 0.184 vs. 1.564 ± 0.130, P < 0.001) expression than NP tissues. E2-stimulated cells had higher AQP5 expression (1.298 ± 0.058 vs. 1.085 ± 0.104, P = 0.049), increased cell proliferation (1.510 ± 0.089 vs.1.000 ± 0.038, P < 0.001), and EMT (TGF-β1 concentration: 0.352 ± 0.021 ng/mL vs. 0.125 ± 0.014 ng/mL, P < 0.001; vimentin: 1.641 ± 0.120 vs. 0.188 ± 0.020, P = 0.002; E-cadherin: 0.075 ± 0.030 vs. 0.843 ± 0.046, P < 0.001) than controls. E2-stimulated cells with AQP5 knockdown exhibited decreased EMT (TGF-β1 concentration: 0.223 ± 0.041 ng/mL vs. 0.352 ± 0.021 ng/mL, P= 0.016; vimentin: 0.675 ± 0.056 vs. 1.641 ± 0.120, P = 0.001; E-cadherin: 0.159 ± 0.037 vs. 0.075 ± 0.030, P = 0.040) than E2-stimulated cells with non-related small interfering RNA (siRNA). Conclusion::Our findings suggest that estrogen induces BPH possibly by promoting AQP5 expression. Hence, AQP5 might be a novel target for modulating EMT in prostate epithelial cells.
9.Detection of BK virus infection in renal transplant recipents and clinical application
Zhong-Biao WU ; Guo-Bing LIN ; Ai-Ping ZENG ; Zhen-Qian CHEN ; Jun CHEN ; Min-Qiao ZHENG ; Guo-Wei TU ; Rui-Ming RONG
Chinese Journal of Experimental and Clinical Virology 2010;24(5):367-369
Objective To study the detection methods of BK virus infection in kidney transplant recipients, and to explore the clinical application. Methods 132 cases of renal transplant recipients were undertaken BK virus detection including presence of decoy cells in urinary sediment, urine and serum BKVDNA to demonstrate the BK virus replication. Result Among 132 cases of renal transplant recipients,urinary decoy cell was found in 37 (28.0%)patients and the median time was 12 months after surgery. 32(24. 2% )patients were diagnosed as BK viruria at a median of 11 months after surgery, and 16( 12. 1% )recipients were diagnosed as BK viremia at a median of 15 months after surgery, 5 patients with BK viruria were diagnosed as BK virus associated nephropathy according to allograft biopsy. Conclusion To make early diagnosis of BK virus infection, detection of urine decoy cells and BKV-DNA in urine and plasma sample is important,which provides an important basis for the prevention of BK virus associated nephropathy.
10.Activation of necroptosis in a rat model of acute respiratory distress syndrome induced by oleic acid.
Long PAN ; Dun-Chen YAO ; Yu-Zhong YU ; Bing-Jun CHEN ; Sheng-Jie LI ; Gui-He HU ; Chang XI ; Zi-Hui WANG ; Jian-Hua LI ; Jie LONG ; Yong-Sheng TU
Acta Physiologica Sinica 2016;68(5):661-668
The present study was aimed to investigate the role of necroptosis in the pathogenesis of acute respiratory distress syndrome (ARDS). The rat model of ARDS was induced by intravenous injection of oleic acid (OA), and observed for 4 h. The lung injury was evaluated by arterial blood gas, lung wet-dry weight ratio (W/D) and histological analyses. Simultaneously, bronchoalveolar lavage fluid (BALF) was collected for total and differential cell analysis and total protein determination. Tumor necrosis factor alpha (TNF-α) level in BALF was determined with a rat TNF-α ELISA kit. Expressions of receptor interacting protein kinase 1 (RIPK1), RIPK3 and mixed lineage kinase domain-like protein (MLKL) in lung tissue were determined by Western blot and immunohistochemical staining. The interaction between RIPK1 and RIPK3 was explored by immunoprecipitation. The results showed that, compared with those in control group, total white blood cells count (WBC), polymorphonuclear percentage (PMN%), total protein concentration, TNF-α level in BALF, W/D, and the alveolar-arterial oxygen tension difference (P(A-a)O) in OA group were significantly increased at 4 h after OA injection. Western blot and immunostaining further showed remarkably increased expressions of RIPK1, RIPK3 and MLKL in lung tissue from OA group. Additionally, immunoprecipitation results indicated an enforced interaction between RIPK1 and RIPK3 in OA group. Collectively, the TNF-α level in BALF and the RIPK1-RIPK3-MLKL signaling pathway in lung tissue were found to be upregulated and activated with the process of ARDS. These findings implicate that RIPK1/RIPK3-mediated necroptosis plays a possible role in the pathogenesis of ARDS, which may provide a new idea to develop novel drugs for the therapy of ARDS.
Acute Disease
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Animals
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Bronchoalveolar Lavage Fluid
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Disease Models, Animal
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Lung Diseases
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Necrosis
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Oleic Acid
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Rats
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Receptor-Interacting Protein Serine-Threonine Kinases
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Respiration Disorders
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Signal Transduction
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Tumor Necrosis Factor-alpha