Aim To study the effect of Vitamin C on growth and apoptosis of human cervical cancer cell line HeLa and its mechanisms. Methods HeLa cell line was treated with VitC. Cell proliferation was evaluated with MTT assay. The distribution of cell cycle,the rate of apoptosis and the expression of HPV18 E6,p53,Bax and Bcl-2 protein of the heLa cells, were observed using flow cytometer respectively. TRAP-PCR-ELISA assay was used to evaluate telomerase activity. Results Vitamin C arrested the HeLa cell line in G_0/G_1 phase in a dose and time-dependent manner, and significantly inhibited HeLa cells growth and increased the rate of apoptosis.Expression of HPV18 E6 and Bcl-2 proteins, was down regulated expression of p53 and Bax proteins was upregulated, and the telomerase activity was reduced. Conclusion Vitamin C can significantly inhibit HeLa cells growth and proliferation,arrest them in G_0/G_1 phase and induce their apoptosis.The reason of these changes is the decrease of E6 gene′s expression, and the succedent changes of proteins′ expression in cancer cells,including the downregulated expression of HPV18 E6 and Bcl-2 proteins, upregulated expression of p53 and Bax proteins and reduced telomerae activity.

Objective To investigate the effects of human umbilical mesenchymal stem cells (hUCMSCs) on Wistar rat models of stress urinary incontinence and its possible mechanism.Methods hUCMSCs cells were separated and extracted from human umbilical cord of cesarean delivery of full-term pregnancy women.The models of stress urinary incontinence (SUI) of Wistar rats were established by imitating delivery damage and ovary removing surgery,and then randomly divided into three groups:model group,transforming growth factor-β1 (TGF-β31) group,hUCMSCs group,and normal rats as normal group;and every group included ten rats.Maximal bladder capacity (MBC),leak point pressure (LPP),abdominal leak point pressure (ALPP),and sneezing experiment of rats including normal rats,model rats and model rats accepted TGF-β1 or hUCMSCs treatment were examined.When the treatment accomplished,the rats were killed and urethral sphincter were separated and examined by hematoxylin eosin (HE) staining and the proteins of troponin Ⅰ (TnI),troponin C (TnC),troponin T (TnT),tropomyosin (Tm),actin (AT),myosin heavy chain (MHC) and myosin light chain (MLC) of urethral sphincter was detected with Western blot method.Results The cells of hUCMSCs were extracted and authenticated,SUI models of Wistar rats were successfully established and authenticated.Compared to model group and TGF-β1 group,the MBC,LPP and ALPP of hUCMSCs group dramatically improved (P ＜ 0.05),while the positive rate of sneezing experiment significantly dropped (P ＜ 0.05).HE staining showed urethral sphincter became attenuation,fracture,sparse and disorder,simultaneously the proteins of TnI,TnC,TnT,Tm,MHC and MLC of urethral sphincter reduced (P ＜ 0.05).While the damaged urethral sphincter basically restored the normal structure and the proteins of TnI,TnC,TnT,Tm,MHC and MLC of urethral sphincter up-regulated through hUCMSCs treatment (P ＜ 0.05).Conclusions The treatment of hUCMSCs translation could observably improve the clinic symptoms of SUI Wistar rat models,and its mechanism may be related to enhancement of the protein expression of TnI,TnC,TnT,Tm,MHC and MLC of urethral sphincter,and were involved in the rehabilitation and reconstruction of urethral sphincter.

Objectives To study the clinical effects and therapeutic mechanisms of the application of Chinese traditional medicine,the Decoction of Jie Du Hua Yu Tang,in the treatment of psoriasis.Meth-ods The clinical effects were observed in50patients with psoriasis vulgaris,to whom the Decoction of Jie Du Hua Yu Tang was given.The mice were used as experimental models whose tail scale epidermis was nat-urally lacking granular layer,similar to psoriatic epidermis.The mice were fed with Jie Du Hua Yu Tang and the effects on the formation of granular layer in their tail scale epidermis were noticed.Results The clinical cure rate was42%and the total response rate was90%with the Jie Du Hua Yu Tang.There was a signifi-cantly strong promoting effect on the formation of granular layer in mouse tail scale epidermis with the use of Jie Du Hua Yu Tang,in comparison with the control group.Conclusions The Decoction of Jie Du Hua Yu Tang is effective for the treatment of psoriasis vulgaris.The decoction could promote the formation of granu-lar layer in mouse tail scale epidermis.The therapeutic mechanism of the decoction may be related to the inhibition of proliferation of epidermal cells.

Tangcao pill is commonly applied in adjuvant and even alternative therapy for patients with AIDS. However, the herb contains complex ingredients, but with unknown effect against anti-HIV drug and unknown function. Because CYP450 emzyme is the main metabolic enzymes of the drug, it is of important significance to study the regulation of CYP450 enzymes before and after the combined administration of Tangcao pill and EFV. Proteomics, due to its high throughout and high sensitivity, has been widely applied in CYP450 enzyme study. In this paper, liver microsomes were separated through differential centrifugation. Their proteins were separated through SDS-PAGE. The three protein bands that CYP450 enzymes were located were cut and identified by liquid chromatography tandem mass spectrometry. Totally 16 CYP450 isoenzymes were identified. Furthermore, in order to make a quantitative analysis on the effect of tang herb on CYP450 emzyme, the multiple reaction monitoring (MRM) technology based on MS was adopted. The CYP2C11 was selected based on the results of the mass spectrum identification of proteins. The characteristic polypeptides were obtained through searching Expasy blast database. The m/z of the fragment ions was less than 800. In the paper, the m/z of ion pairs of CYP2C11 were 711.5/232.1, 711.5/319.2, 711.5/466.2 and 711.5/595.3, and the m/z of ESAT-6 (internal standard, IS) were 735.5/215.3, 735.5/389.3, 735.5/460.3 and 735.5/524.3. The relative peak (analyte/IS) area was adopted for the relative quantitative analysis. Compared with the EFV single administration group, the EFV and Tangcao pill combined administration group showed a 1.6-fold increase in CYP2C11. The results of the paper indicated that Tangcao pill may affect drug metabolism by regulating metabolic enzymes such as CYP2C11, but the specific mechanism still unknown.
Animals ; Cytochrome P-450 Enzyme System ; chemistry ; genetics ; metabolism ; Drugs, Chinese Herbal ; administration & dosage ; Electrophoresis, Polyacrylamide Gel ; Male ; Microsomes, Liver ; chemistry ; drug effects ; enzymology ; Proteomics ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo observe the rate of efficacy and adverse drug reaction of non-steroidal anti-inflammatory drugs (NSAIDs) in the population with osteoarthritis and rheumatoid arthritis, based on available clinical data.

METHODSUsing Meta analysis to evaluate the data of effect and safety profile of NSAIDs from 19 articles on randomized clinical trials published from 1990 to 2001 in Chinese journals. The total number of patients enrolled for evaluation on rates of effectiveness and adverse drug reaction were 1 732 and 2 925, respectively.

RESULTSData on the effect and safety were comparatively heterogeneous among different kinds of NSAIDs. The effective rates (95% CI) were as follows: nabunetone, 66.7% (61.9% - 71.4%); meloxicam, 68.4% (59.2% - 77.6%); naproxen, 64.5% (59.8% - 69.1%); nimesulide, 79.8% (75.7% - 84.0%); ibuprofen, 77.2% (70.7% - 83.8%); diclofenac, 77.1% (69.2% - 85.0%); oxaprozin, 65.8% (59.5% - 72.0%). Rates of adverse drug reaction (95% CI) were as follows: nabunetone, 16.3% (12.5% - 20.0%); meloxicam, 10.2% (4.2% - 16.2%); naproxen, 29.2% (24.8% - 33.6%); nimesulide, 20.2% (16.0% - 24.3%); ibuprofen, 16.7% (14.7% - 18.8%); diclofenac, 19.3% (11.9% - 26.7%); oxaprozin, 12.7% (8.9% - 16.7%) respectively.

CONCLUSIONThe rates of effect and adverse reaction on patients having osteoarthritis and rheumatoid arthritis with NSAIDs treatment would largely depend on the drugs being used. Within 2 - 8 weeks of treatment, the effective rate and rate of adverse drug reaction with commonly used NSAIDs as nabumeton, meloxicam, etc., were 59.2% - 85.0% and 4.2% - 33.6%, respectively.

Anti-Inflammatory Agents, Non-Steroidal ; adverse effects ; therapeutic use ; Arthritis, Rheumatoid ; drug therapy ; Butanones ; adverse effects ; therapeutic use ; China ; Diclofenac ; adverse effects ; therapeutic use ; Humans ; Ibuprofen ; adverse effects ; therapeutic use ; Naproxen ; adverse effects ; therapeutic use ; Osteoarthritis ; drug therapy ; Propionates ; adverse effects ; therapeutic use ; Randomized Controlled Trials as Topic ; Sulfonamides ; adverse effects ; therapeutic use ; Thiazines ; adverse effects ; therapeutic use ; Thiazoles ; adverse effects ; therapeutic use

This study is to investigate the effects of aqueous extract of Schisandra chinensis Baill (WWZ), kadsurin, schisandrin A, schisandrin B and schisandrol B on rat hepatic CYP3A. Rats received a daily gavage of aqueous extract of WWZ for different times. The livers were harvested after gavage and subjected to microsome preparation. Microsomal CYP3A activity was determined by measuring the amount of the metabolite of testosterone (6 beta-hydroxytestosterone) with HPLC. Aqueous extract of WWZ, kadsurin and schisandrin A were incubated with microsomes obtained from rat. Microsomal CYP3A activity was determined by HPLC. Primary hepatocytes were separated and extracted from rat, then were treated with aqueous extract of WWZ, schisandrin A, schisandrin B and schisandrol B. Then, the expression of CYP3A1 mRNA was analyzed by RT-PCR. As for the in vivo assay, aqueous extract of WWZ significantly inhibited the enzyme activity of CYP3A after 12 h gavage. The inhibitory effect was converted to inductive effect after 3-day gavage. Aqueous extract of WWZ could induce the enzyme activity of CYP3A after 6-day gavage. Aqueous extract of WWZ and kadsurin showed a dose-dependent inhibition of CYP3A (IC50 of 487.8 microg mL(-1) and 6.2 micromol L(-1), separately). In rat primary hepatocytes, aqueous extract of WWZ (2.5 mg mL(-1)), schisandrin A (0.1 micromol L(-1)), schisandrin B (0.1 micromol L(-1)) and schisandrol B (10 micromol L(-1)) increased significantly the expression of CYP3A1 mRNA by 23%, 55%, 42% and 27%, respectively. Aqueous extract of WWZ could show dual effect on the enzyme activity of CYP3A in rat in vivo. Meanwhile, kadsurin showed a dose-dependent inhibition of the enzyme activity of hepatic CYP3A in vitro. And schisandrin A, schisandrin B and schisandrol B showed significant inductive effect on the expression of rat CYP3A1 mRNA.
Animals ; Cyclooctanes ; administration & dosage ; isolation & purification ; pharmacology ; Cytochrome P-450 CYP3A ; genetics ; metabolism ; Dioxoles ; administration & dosage ; isolation & purification ; pharmacology ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; pharmacology ; Hepatocytes ; drug effects ; enzymology ; Inhibitory Concentration 50 ; Lignans ; administration & dosage ; isolation & purification ; pharmacology ; Male ; Microsomes, Liver ; enzymology ; Plants, Medicinal ; chemistry ; Polycyclic Compounds ; administration & dosage ; isolation & purification ; pharmacology ; RNA, Messenger ; metabolism ; Rats ; Rats, Sprague-Dawley ; Schisandra ; chemistry

Objective To induce mouse embryonic stem(ES)cells to differentiate into insulin-secreting cells by means of a 5-step model system.Methods E14.1 mouse ES cells were cultured in the presence of leukemia inhibitory factor(LIF)for 2 days(step 1),then the cells were cultured in hanging drops to form embryonic bodies(EBs)and the resulting EBs were cultured in suspension for 6 days in the presence of basic fibroblast growth factor bFGF(step 2).Subsequently the EBs were cultured in the medium containing glucagon- like peptide 1(GLP-1),hepatocyte growth factor(HGF),nerve growth factor(NGF)and nicotinamide for 10 days(step 3).After that,the EBs were dissociated into single cells,and the cells were cultured in monolayer in the presence of GLP-1,betacellulin,activin A,bFGF and nicotinamide for 10 days(step 4).Finally,the cells were cultured in low-glucose medium containing nicotinamide for 4 days(step 5).Insulin and some other islet- related genes expressions were investigated using RT-PCR and insulin expression was also investigated by DTZ- staining and immunohistochemistry.The percentage of insulin-secreting cells was evaluated by flowcytometry and insulin concentrations were measured by RIA.Results mRNA expression of insulin became visible at step 3 and more evident at step 5.Additionally,at step 5,mRNAs of glucagon,somatostatin,pancreatic polypeptide(PP), pancreatic duodenal homeobox 1(PDX-1),beta-cell E box transactivator 2(Beta2)and neurogenin 3(Ngn3) were detected.DTZ-staining positive cells and insulin immunohistochemical staining positive cells were observed. The percentage of insulin-positive cells was(24.0?2.5)%(n=6).In the presence of 5.6 mmol/L and 25 mmol/L glucose,insulin concentrations were(0.05?0.01)?g/L and(0.13?0.02)?g/L respectively(n= 6).Conclusion E14.1 mouse ES cells can be induced to differentiate into insulin-secreting cells by the 5-step model system.Insulin-secreting cells can release insulin into culture medium when treated with glucose,and insulin concentrations increase with rising concentration of glucose.

Objective To explore the expression and mechanism of the hypoxia inducible factor (HIF) and their downstream responsive genes BNIP3, VEGF in gastric adenocarcinoma, and to investigate their clinical significance. Methods SP immunohistochemical method was used to detect the expression of HIF-1α, HIF-2α, BNIP 3 and VEGF in the tissue samples from 70 gastric adenocarcinoma patients. Results Compared with the normal tissue, gastric adenocarcinoma had significantly higher positive expression rate of HIF-1α(61.4% vs. 8.3%), HIF-2α(37.1% vs.0), BNIP3(51.4% vs.0) and VEGF(62.9% vs. 0). The expression of BNIP3 and VEGF was positively correlated with HIF-1α expression. The expressions of HIF-1α and VEGF was significantly and positively correlated with lymph node metastasis, while BNIP3 expression was negatively correlated with that. Conclusion Accumulation of hypoxia inducible factors regulate the expression and function of its downstream target genes, which may play a critical role in the carcinogenesis of gastric adenocarcinoma. VEGF expression is found to be related to poor biological behavior, but BNIP3 expression may suggest better prognosis of gastric adenocarcinoma.

Objective To explore the expression and mechanism of the hypoxia inducible factor (HIF) and their downstream responsive genes BNIP3, VEGF in gastric adenocarcinoma, and to investigate their clinical significance. Methods SP immunohistochemical method was used to detect the expression of HIF-1α, HIF-2α, BNIP 3 and VEGF in the tissue samples from 70 gastric adenocarcinoma patients. Results Compared with the normal tissue, gastric adenocarcinoma had significantly higher positive expression rate of HIF-1α(61.4% vs. 8.3%), HIF-2α(37.1% vs.0), BNIP3(51.4% vs.0) and VEGF(62.9% vs. 0). The expression of BNIP3 and VEGF was positively correlated with HIF-1α expression. The expressions of HIF-1α and VEGF was significantly and positively correlated with lymph node metastasis, while BNIP3 expression was negatively correlated with that. Conclusion Accumulation of hypoxia inducible factors regulate the expression and function of its downstream target genes, which may play a critical role in the carcinogenesis of gastric adenocarcinoma. VEGF expression is found to be related to poor biological behavior, but BNIP3 expression may suggest better prognosis of gastric adenocarcinoma.