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1.
Article in English | WPRIM | ID: wpr-776624

ABSTRACT

OBJECTIVE@#To evaluate the effect of Zhizhu Kuanzhong Capsules (, ZKC) for functional dyspepsia (FD) through meta-analysis.@*METHODS@#Online databases, including PubMed, EM base, China National Knowledge Infrastructure, Wanfang Data, VIP database and Cochrane Library, were searched for randomized controlled trials (RCTs) of ZKC for FD from the inception to April, 2016. Trials were selected according to inclusion criteria and were evaluated with quality assessment standards in the Cochrane Handbook for Systematic Reviews of Interventions and Jadad scale. RevMan 5.3 and GRADEprofiler 3.6 were used for statistical analysis and evidence quality assessment.@*RESULTS@#Twenty-three trials with 2,496 patients were included and most of them were of poor methodological quality. ZKC alone or ZKC combined with routine Western medicine (WM) showed a better clinical effect rate compared with the control group of WM [odds ratio (OR)=3.32, 95% confidence interval (2.66, 4.15), P<0.00001]. No serious adverse reactions were reported.@*CONCLUSIONS@#ZKC alone or ZKC combined with routine WM could significantly improve the clinical effective rate in the treatment of FD. The quality of the evidence is low, so it is necessary to design multicenter, strictly randomized and double-blind controlled trials with large samples to validate the conclusions.

2.
Article in Chinese | WPRIM | ID: wpr-238596

ABSTRACT

<p><b>OBJECTIVE</b>To observe and compare the effects of Hanfangji Compound and IFN-gamma on expressions of transthyretin (TTR) , inter-alpha inhibitor H1 (ITIH1) and serpin peptidase inhibitor clade F member 2 (SERPINF2) of hepatic stellate cells (HSC-T6).</p><p><b>METHOD</b>Hanfangji Compound and IFN-gammaof different concentrations were used in hepatic stellate cell-T6 (HSC-T6) for 48 h. Flow cytometer was used to detect the effects of Hanfangji Compound and IFN-gamma on HSC proliferation. RT-PCR method was adopted to detect mRNA expressions of TFR, ITIH1 and SERPINF2. TTR, ITIH1 and SERPINF2 secretions were detected by ELISA. The protein localizations of TTR, ITIH1 and SERPINF2 were examined by immune fluorescence. The protein expression of TfR and ITIHI were determined by Western blot.</p><p><b>RESULT</b>After Hanfangji Compound and IFN-gamma were adopted in HSC-T6, compared with the control group, the cell proliferation was inhibited obviously (P < 0. 05) , protein expressions of TTR, ITIH1 and SERPINF2 and mRNA expression increased significantly, with certain correlation with concentrations of Hanfangji Compound. The 2. 5 g L-I Hanfangji Compound group was superior to the IFN-gamma group (P <0. 05).</p><p><b>CONCLUSION</b>Hanfangji Compound can inhibit HSC proliferation, upregulated TTR, ITIH1 and SERPINF2 proteins and mRNA expression, which may be one of mechanisms of anti-hepatic fibrosis of Hanfangji Compound.</p>


Subject(s)
Alpha-Globulins , Genetics , Metabolism , Blotting, Western , Cell Line, Tumor , Cell Proliferation , Drugs, Chinese Herbal , Pharmacology , Enzyme-Linked Immunosorbent Assay , Hepatic Stellate Cells , Cell Biology , Metabolism , Humans , Receptors, Albumin , Genetics , Metabolism , alpha-2-Antiplasmin , Genetics , Metabolism
3.
Chinese Journal of Hepatology ; (12): 614-618, 2013.
Article in Chinese | WPRIM | ID: wpr-278029

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the effects and related mechanisms of hepatitis B virus X (HBx) protein on cell cycle and growth in hepatocellular carcinoma.</p><p><b>METHODS</b>A human hepatocyte HepG2 cell line stably expressing a green fluorescent protein (GFP)-tagged HBx (HepG2/GFP-HBx cells) was used for the experiment, and HepG2 parental and HepG2/GFP cells was used as the controls. Effect of HBx on cell growth was evaluated by the MTT cell proliferation assay and on cell cycle progression by flow cytometry analysis of cells with or without treatment with 5-aza-2'-deoxycytidine (5-Aza-CdR; 5 pmol/L). Effect of HBx expression on promoter methylation status of the p16INK4A tumor-suppressor gene was detected by methylation-specific polymerase chain reaction and on p16 protein level was analyzed with western blotting.</p><p><b>RESULTS</b>The HepG2/GFP-HBx cells showed significantly higher cell proliferation at 72 hrs of culture (3.225+/-0.038 A490) than either control (HepG2: 2.012+/-0.022 A490, t = -46.86, P less than 0.001; HepG2/GFP: 2.038+/-0.029 A490, t = 42.51, P less than 0.001). The HepG2/GFP-HBx cells also showed significantly lower proportion of cells in the G0/G1 phase (16.45%+/-0.45%) than either control (HepG2: 44.81%+/-1.36%, t = -34.202, P less than 0.001; HepG2/GFP: 42.76%+/-1.58%, t = -28.88, P less than 0.001). However, 5-Aza-CdR treatment did lead to a significant amount of HepG2/GFP-HBx cells being arrested in the G0/G1 phase (33.25%+/-0.79%, t = 31.85, P less than 0.001). The p16INK4A promoter was methylated in the HepG2/GFP-HBx cells, and became demethylation after treatment with 5-Aza-CdR. However, no methylation of p16INK4A promoter was observed in both HepG2 and HepG2/GFP cells. The p16 protein level was significantly lower in the HepG2/GFP-HBx (vs. HepG2 and HepG2/GFP cells) and this level increased after treatment with 5-Aza-CdR.</p><p><b>CONCLUSION</b>HBx protein promotes hepatocellular carcinoma cell cycle progression and growth by shortening the G0/G1 phase, and the underlying mechanism may involve inducing p16INK4A promoter methylation and downregulating p16 protein expression.</p>


Subject(s)
Carcinoma, Hepatocellular , Metabolism , Pathology , Cell Cycle , Cell Proliferation , Cyclin-Dependent Kinase Inhibitor p16 , Genetics , Metabolism , Gene Expression Regulation, Neoplastic , Genes, p16 , Hep G2 Cells , Hepatitis B virus , Metabolism , Humans , Liver Neoplasms , Metabolism , Pathology , Promoter Regions, Genetic , Trans-Activators , Pharmacology
4.
Article in Chinese | WPRIM | ID: wpr-301397

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the expression of peripheral blood gammadelta T cells/CD4 CD25+ regulatory T cells(Treg) and cytokines interleukin 17 (IL-17) and transforming growth factor beta1 (TGF-beta1) in patients with allergic rhinitis.</p><p><b>METHODS</b>From March 2012 to July 2012, 32 patients with allergic rhinitis (AR group) and 20 healthy control subjects (control group) were collected. The expression of peripheral blood gammadelta T cells/Treg cells were measured by flow cytometry and the levels of IL-17 and TGF-beta1 were evaluated by ELISA. SPSS 16.0 software was used to analyze the data.</p><p><b>RESULTS</b>The percentages of gammadeltaT cells in AR group were (13.30 +/- 8.62)%, which was significantly higher (t = 5.18, P < 0.01) than those in control group (5.18 +/- 1.86)%. The percentages of Treg cells in AR group were (1.75 +/- 0.56)%, which were significantly lower (t = 7.46, P < 0. 01) than those in control group (4.76 +/- 1.74)%. The IL-17 levels in AR group were (668.55 +/- 45.15) pg/ml, which were also significantly higher (t = 8.97, P < 0.01) than those in control group (573.53 +/- 17.42) pg/ml. The TGF-beta1 levels in AR group were (0.34 +/- 0.04) pg/ml, which were also significantly lower (t = 9.51, P < 0.01) than those in control group (0.49 +/- 0.06) pg/ml. There was a negative correlation between the percentages of gammadelta T cells and Treg cells (r = -0.561, P < 0.01). There was a negative correlation between the percentages of gammadelta T cells and TGF-beta1 levels (r = -0.622, P < 0.01). A positive correlation was shown between the percentages of gammadelta T cells and IL-17 levels in AR (r = 0.469, P < 0.01). A positive correlation was shown between the percentages of Treg cells and TGF-beta1 levels in AR (r = 0.738, P < 0.01). There was no correlation between IL-17 levels and the percentages of Treg cells or TGF-beta1 levels (r value was -0.111, -0.196, all P > 0.05).</p><p><b>CONCLUSION</b>There are imbalances of gammadelta T and Treg cells in peripheral blood of patients with allergic rhinitis. gammadelta T cells may be the main cell to produce IL-17, which may play an important role in allergic rhinitis.</p>


Subject(s)
Enzyme-Linked Immunosorbent Assay , Flow Cytometry , Humans , Interleukin-17 , Metabolism , Rhinitis, Allergic , Allergy and Immunology , Metabolism , T-Lymphocytes, Regulatory , Transforming Growth Factor beta1 , Metabolism
5.
Acta Pharmaceutica Sinica ; (12): 365-370, 2010.
Article in Chinese | WPRIM | ID: wpr-250578

ABSTRACT

To develop and validate a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the quantification of epirubicin hydrochloride (EPI) in rat plasma, daunorubicin hydrochloride was used as internal standard. The plasma samples were deproteinated with methanol, and separation was performed on a reversed-phase CAPCELL PAK C18 column (3.0 mm x 50 mm, 3 microm). The mobile phase contained methanol-0.1% formic acid (80:20). Detection was carried out by multiple reaction monitoring on a HP1200-6410 QQQ LC/MS system. Different preparations of EPI solution, EPI-LIP (EPI-liposome) and EPI-LTSL (EPI-thermosensitive liposome) was administered in rats by i.v with the same dosage (12 mg kg(-1)). The pharmacokinetic model and parameters were fitted and calculated by the DAS ver2.0 software. The calibration curve was linear in the range of 0.01-50 microg mL(-1). The limit of quantification was 0.01 microg mL(-1). RSDs of intra- and interbatch precisions were all less than 11.9%. The average extract recovery was 89.3% and 92.1%, respectively. The pharmacokinetics of EPI in rats with all preparations were fitted to three compartments, which all fast distributed and slowly eliminated. The t1/2 alpha, t1/2 beta, t1/2 gamma, AUC(0-infinity), and MRT(0-infinity) of EPI-LTSL group were 7.5, 1.3, 12.6, 12.9, 3.7 times those of EPI solution group; and 1.6, 1.4, 12.3, 2.9, 2.6 times those of EPI-LIP group. Moreover, the CL of the latter two groups was about 13.4 times of the former EPI-LTSL group. EPI-LTSL can significantly improve AUC and prolong the circulation time of EPI in rat plasma.


Subject(s)
Animals , Antibiotics, Antineoplastic , Blood , Pharmacokinetics , Area Under Curve , Chromatography, Liquid , Drug Carriers , Epirubicin , Blood , Pharmacokinetics , Liposomes , Blood , Pharmacokinetics , Male , Rats , Rats, Sprague-Dawley , Reproducibility of Results , Sensitivity and Specificity , Tandem Mass Spectrometry
6.
Article in Chinese | WPRIM | ID: wpr-330845

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the effect of hepatitis B virus X protein (HBx) on adriamycin-induced apoptosis of hepatocellular carcinoma cells and the expressions of p53 and PTEN.</p><p><b>METHODS</b>HepG2, HepG2/GFP, and HepG2/GFP-HBx cells were treated with adriamycin (2.5 microg/ml), and the apoptotic cell death was determined by observing the morphological changes and flow cytometry. The expressions of p53 and PTEN mRNA in the 3 cells were detected by RT-PCR, and the expressions of p53 and PTEN protein were analyzed by Western blotting.</p><p><b>RESULTS</b>Adriamycin induced significant cell death in HepG2 and HepG2/GFP cells, which became rounded, shrunk, and detached after the treatment; but no significant cell death occurred in HepG2/GFP-HBx cells. Flow cytometry analysis showed that the apoptotic rate was significantly lower in HepG2/GFP-HBx cells (3.94%) than in HepG2 (59.03%) and HepG2/GFP cells (61.38%) at 36 h after the treatment (P<0.001), while no significant difference was observed between HepG2/GFP-HBx (3.94%) and the control cells (2.12%, 2.78%, and 2.55%) (P>0.05). RT-PCR showed lowered expression of PTEN mRNA in HepG2/GFP-HBx cells as compared to that in HepG2 and HepG2/GFP cells, while no significant difference was noted in p53 mRNA. Western blot analysis showed that PTEN protein decreased while p53 protein remain unchanged in HepG2/GFP-HBx cells.</p><p><b>CONCLUSION</b>HBx suppresses adriamycin-induced apoptosis of HepG2 cells and PTEN expression. The inhibitory effect of HBx on the cell apoptosis may be related to the inhibition of p53-PTEN pathway.</p>


Subject(s)
Apoptosis , Carcinoma, Hepatocellular , Metabolism , Pathology , Doxorubicin , Pharmacology , Hep G2 Cells , Humans , Liver Neoplasms , Metabolism , Pathology , PTEN Phosphohydrolase , Metabolism , Trans-Activators , Metabolism , Tumor Suppressor Protein p53 , Metabolism
7.
Chinese Medical Journal ; (24): 1784-1789, 2009.
Article in English | WPRIM | ID: wpr-240797

ABSTRACT

<p><b>BACKGROUND</b>The role of B-cell remains an enigma in the pathogenesis of ankylosing spondylitis (AS). This study aimed to investigate the distributions of B-cells and subsets in peripheral blood of AS patients and observe their changes in etanercept-treated AS patents.</p><p><b>METHODS</b>We detected the proportions of CD19(+) B-cell, naive B-cell (CD19(+)CD27-), memory B-cell (CD19(+)CD27dim) and plasmablast (CD19(+)CD27high) in peripheral blood of 66 patients with AS (39 at active stage, 27 at stable stage; 35 patients with peripheral joint involvement, 31 patients with axial involvement alone), 30 patients with rheumatoid arthritis (RA) and 30 healthy volunteers using flow cytometry. And then we observed the changes of the above indexes of 39 active AS patients treated with etanercept in a randomized, double-blind, placebo-controlled trial.</p><p><b>RESULTS</b>(1) Percentages of CD19(+) B-cells in active or peripheral joint involvement AS patients increased more obviously than those in stable or axial involvement alone AS patients (both P = 0.001), and percentage of CD19(+)CD27high B-cells in AS patients with peripheral joint involvement was significantly higher than that in cases with axial involvement alone or healthy volunteers (P = 0.005 and 0.006, respectively); (2) The percentage of CD19(+) B-cells in AS patients was positively correlated with Bath Ankylosing Spondylitis Disease Activity Index (BASDAI) scores, Patient's Global Assessment (PGA) scores, total back pain scores and nocturnal back pain scores (r = 0.270, 0.255, 0.251 and 0.266, P = 0.029, 0.039, 0.042 and 0.031, respectively); (3) At week 6 and week 12, there were no statistical differences of the percentages of B-cells and subsets between etanercept group and placebo group of AS patients (P > 0.05); the percentage of CD19(+) B-cells in etanercept group was higher than that in healthy volunteers at week 12 (t = 3.320, P = 0.003).</p><p><b>CONCLUSIONS</b>Misbalance is present in B-cells and some subsets in peripheral blood of active AS patients with peripheral joint involved. B-cells might play an important role in the pathogenesis of AS patients. The high percentage of CD19(+) B-cells in active AS patients cannot be down-regulated after 12-week etanercept treatment.</p>


Subject(s)
Adolescent , Adult , Antigens, CD19 , Allergy and Immunology , B-Lymphocytes , Allergy and Immunology , Etanercept , Female , Flow Cytometry , Humans , Immunoglobulin G , Pharmacology , Therapeutic Uses , Immunosuppressive Agents , Pharmacology , Therapeutic Uses , Male , Middle Aged , Receptors, Tumor Necrosis Factor , Therapeutic Uses , Spondylitis, Ankylosing , Drug Therapy , Allergy and Immunology , Tumor Necrosis Factor Receptor Superfamily, Member 7 , Allergy and Immunology , Young Adult
8.
Chinese Journal of Hepatology ; (12): 25-28, 2008.
Article in Chinese | WPRIM | ID: wpr-277617

ABSTRACT

<p><b>OBJECTIVES</b>To investigate the effect of hepatitis B virus X protein (HBx) on adriamycin-induced apoptosis of hepatocellular carcinoma cells.</p><p><b>METHODS</b>HBx gene fragment was amplified from subtype adr HBV plasmid by PCR, and inserted into Hind III and Kpn I sites of green fluorescent protein (GFP) eukaryotic expression vector pEGFP-C1 to construct recombinant pGFP/HBx. The pEGFP-C1 and pGFP-HBx were introduced into HepG2 cells by Lipofectamine 2000 to obtain HepG2 cells expressing GFP. GFP-HBx fusion protein was selected using G418. The expression of HBx gene was demonstrated by RT-PCR analysis. HepG2, HepG2/GFP and HepG2/GFP-HBx cells were treated with adriamycin (2.5 microg/ml), and apoptosis of the cells was determined by their morphological changes, trypan blue exclusion, and flow cytometry analysis.</p><p><b>RESULTS</b>Under a fluorescence microscope, visible expression of GFP and GFP-HBx fusion proteins were observed in HepG2/GFP and HepG2/GFP-HBx cells, even after growing over 70 generations. RT-PCR analysis showed that HBx gene was expressed in HepG2/GFP-HBx cells. Trypan blue exclusion showed adriamycin induced time-dependent cell death in HepG2 and HepG2/GFP cells while no significant cell death was observed in HepG2/GFP-HBx cells. Flow cytometry analysis showed that apoptosis rates in HepG2/GFP-HBx (3.94%) cells were significantly lower than those in HepG2 (59.03%) and HepG2/GFP cells (61.38%) at 36 hours after the adriamycin treatment (P < 0.01). No significant differences of apoptosis rates of HepG2/GFP-HBx (3.94%) and of the untreated cells (2.12%, 2.78%, 2.55%) (P > 0.05) were observed.</p><p><b>CONCLUSION</b>A HepG2 cell line expressing GFP and GFP-HBx fusion proteins was successfully established. HBV X protein blocks adriamycin-induced apoptosis of these HepG2 cells.</p>


Subject(s)
Apoptosis , Doxorubicin , Pharmacology , Hep G2 Cells , Humans , Plasmids , Trans-Activators , Genetics
9.
Article in English | WPRIM | ID: wpr-282427

ABSTRACT

<p><b>OBJECTIVE</b>To study the effects of gypenoside (Gyp) on the activity of microsomal Na(+), K(+)-ATPase in rat's heart and brain in vitro.</p><p><b>METHODS</b>The microsomal Na(+), K(+)-ATPase was prepared from rat's heart and brain by differential centrifugation. The activity of microsomal Na(+), K(+)-ATPase was assayed by colorimetric technique. Enzyme kinetic analysis method was used to analyze the effect of Gyp on the microsomal Na(+), K(+)-ATPase of rats.</p><p><b>RESULTS</b>Gyp reversibly inhibited the brain and heart's microsomal Na(+), K(+)-ATPase in a concentration-dependent manner, and showed a more potent effect on enzyme in the brain. The IC(50) of Gyp for the heart and brain were 58.79+/-8.05 mg/L and 52.07+/-6.25 mg/L, respectively. The inhibition was enhanced by lowering the Na(+), or K(+) concentrations or increasing the ATP concentration. Enzyme kinetic studies indicated that the inhibitory effect of Gyp on the enzyme is like that of competitive antagonist of Na(+), the counter-competitive inhibitor for the substrate ATP, and the mixed-type inhibitor for K(+).</p><p><b>CONCLUSION</b>Gyp displays its cardiotonic and central inhibitory effects by way of inhibiting heart and brain's microsomal Na(+), K(+)-ATPase activities in rats.</p>


Subject(s)
Adenosine Triphosphate , Pharmacology , Animals , Brain , Enzyme Inhibitors , Pharmacology , Gynostemma , Kinetics , Male , Myocardium , Plant Extracts , Pharmacology , Rats , Rats, Wistar , Sodium-Potassium-Exchanging ATPase
10.
Chinese Journal of Hepatology ; (12): 721-724, 2006.
Article in Chinese | WPRIM | ID: wpr-260616

ABSTRACT

<p><b>OBJECTIVE</b>To detect HBV antigen specific cytotoxic T lymphocyte (CTL) changes in patients during acute flare-ups and to study their association with flare-ups and aggravations into grave hepatitis by quantitative analysis of HLA-A2* restricted HBcAg-specific CTL cells.</p><p><b>METHODS</b>The frequency of HBcAg-specific CTL cells in the peripheral blood mononuclear cells (PBMC) from 29 patients with persistent infection with HBV were quantified by flow cytometry using one HLA-A2*HBV peptide pentamers complex (Pro5TM MHC Pentamers).</p><p><b>RESULTS</b>There was a statistical difference of HBcAg specific CTLs between the patients with acute exacerbations (1.4%+/-0.8%) and the patients with immune tolerance (0.6%+/-0.4%) (t = 2.180, P = 0.01-0.05); There was no significant difference between the grave hepatitis group (1.3%+/-1.0%) and the chronic hepatitis group (1.4%+/-0.8%) regarding frequencies of antigen specific CTL (t = 0.215, P = 0.833-0.05). The level of antigen specific CTLs in PBMC in the 6 cases of chronic hepatitis B with acute exacerbations maintained a relatively high level (more than 0.7%) within the 12 week follow-up period.</p><p><b>CONCLUSION</b>HBcAg-specific CTLs may play an important role in hepatic flare-ups in patients with chronic HBV infection, but there was no direct relationship between antigen- specific CTLs and grave hepatitis.</p>


Subject(s)
Adult , Female , HLA-A2 Antigen , Allergy and Immunology , Hepatitis B Core Antigens , Allergy and Immunology , Hepatitis B virus , Allergy and Immunology , Hepatitis B, Chronic , Allergy and Immunology , Virology , Humans , Male , Middle Aged , T-Lymphocytes, Cytotoxic , Allergy and Immunology , Viral Load , Young Adult
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