Purpose:To investigate the effect of replication defective retroviral vectors carried sense or antisense TGF?1 fragment on the cell cycle regulation and proliferation of human bladder cancer. Methods:The replication defective retroviral vectors that integrated sense or antisense bioactive fragment of transforming growth factor?1 were constructed,and named as pRevT? and pRevT?-AS respectively. The influence of each vector on the cell proliferation,clone-formation and alteration of cell cycle of bladder cancer cell line EJ were observed in vitro.Results:The titre of pRevT? and pRevT?-AS were 0.84,0.88?10 5 CFU/ml respectively,the vectors integrated to EJ cells and expressed efficiently. Inhibition TGF?1 gene expression reduced proliferation and clone-formation rates of EJ cells. The G 0 /G 1 stage ratios in the antisense TGF?1-transfected EJ cells were increased,simultaneously,the S stage ratios were decreased. Conclusions:The antisense TGF?1 vector can reduce the expression of endogenous TGF?1 in EJ cells,induce G 1 stage arrest and inhibit proliferative growth in vitro.

Objective To investigate the efficacy of sequential add-on of pegylated interferon α-2a (PEGIFN-α-2a) for 48 weeks in chronic hepatitis B (CHB) patients with low serum hepatitis B e antigen (HBeAg) titer after long term adefovir dipivoxil (ADV) monotherapy.Methods This was a randomized,open and prospective clinical trial.Patients who had been treated with ADV for 72 to 144 weeks,with undetectable serum hepatitis B virus (HBV) DNA level,low HBeAg titer (5 S/CO＜ HBeAg＜50 S/CO) and serum hepatitis B surface antigen (HBsAg) ＜5000 IU/mL were included.The patients were categorized into ADV monotherapy group and ADV plus PEGIFN-a-2a combination therapy group by random number table.Patients in ADV group continued ADV monotherapy and patients in combination therapy group added PEGIFN-α-2a to ADV for 48 weeks.After the treatment,efficacy of the two therapies were assessed by comparing the reduction of serum HBeAg reduction,HBeAg loss rate,HBeAg seroconversion rate,and reduction of intrahepatic HBV DNA and HBV covalently closed circular DNA (cccDNA).Pre-and post-treatment results were compared by paired samples t test.Comparison between groups was performed using indepedent samples t test.Comparison of rates between groups was performed using x2 test.Results The trial enrolled 55 CHB patients,and there were 27 patients in ADV monotherapy group,28 patients in combination therapy group.Baseline characteristics including age distribution,sex ratio,alanine aminotransferase (ALT),aspartate aminotransferase (AST),total bilirubin (TBil),serum HBeAg and HBsAg,hepatic HBV DNA and HBV cccDNA were all comparable (all P＞0.05).Twenty-five patients in ADV monotherapy group and 26 patients in combination therapy group completed 48 weeks treatment.HBeAg loss rates and seroconversion rates of combination therapy group were higher than those of ADV monotherapy group (x2 =5.38 and 4.69,respectively,both P＜0.05).HBeAg titers of both groups were significantly lower than those of baseline (t=8.43 and 8.50,respectively,both P＜0.05).The HBeAg titer of combination therapy group was lower than that of monotherapy group (t=5.60,P＜ 0.01).HBV DNA and HBV cccDNA in liver tissue of combination therapy group was (6.934±0.52) lg IU/mg and (5.63±0.54) lg IU/mg post-treatment,respectively,which were both lower than baseline (t＝7.12.6.67,respectively,both P＜0.01).HBV DNA in liver tissue of monotherapy group was (7.09＝0.43) lg IU/mg post-treatment,which was lower than baseline (t=2.67,P=0.02).After treatment,HBV cccDNA in liver tissue of combination therapy group was lower than that of monotherapy group (t =2.87,P=0.00).Conclusions Compared with ADV monotherapy,sequential add-on of PEGIFN-a-2a in combination with ADV can achieve higher serum HBeAg loss rate and seroconversion rate and facilitate the clearance of hepatic HBV DNA and HBV cccDNA in CHB patients with low HBeAg titer after long-term ADV monotherapy.

AIM: To optimize and establish the extraction techn ology of water-soluble polysaccharides from Ulva pertusa. TLC was employed to determine the mainly neutral sugars in the Ulvan. METHODS: The orthogonal test was employed to test the effects of th e four factors including volume of water, temperature of extraction, time of ext raction and pH of water on the yield and total carbohydrate contents of water-s oluble polysaccharides from Ulva pertusa and optimize the extraction technol ogy. Five groups of parallel tests were carried out by the optimum parameters es tablished and the sugar constituents of products were analyzed by TLC. RESULTS: Temperature of extraction and time of extraction have sign ificant effects on the extraction yield of water-soluble polysaccharides from Ulva pertu sa, but, for total carbohydrate content, only temperature of extraction was a significant factor. The mainly neutral sugars were composed of rhamnose, xylose and glucose. CONCLUSION: The optimum extraction technology was A 1B 3C 3D 3 when the yield and total carbohydrate content were considered as preferential pa rameters.

Objective To investigate allergen in children with asthma for prevention and treatment of asthma.Methods 89 patients with asthma were devided into two groups:children group(age≥3) and infant group(age

OBJECTIVE To explore the effect of smearing catheter surface with chloramphenicol for preventing catheter -associated urinary tract infections. METHODS Totally 100 cases of preoperative patients needed for indwelling urethral catheters were randomly grouped, 50 of 100 cases as test group, and the others 50 cases as the control group. Catheters after smearing surface with chloramphenicol were inserted using aseptic technique in the test group, urinary catheters without using chloramphenicol were inserted in the control group according to routine aseptic technique protocol, and then regularly taken out urinary specimen from two groups respectively for microbial culture. RESULTS The observation showed bacterial growth positive rate was 30%, and 66.7%, respectively in the control group, but positive rate was 6.7%, and 30%, respectively in the test group after the seventh day and the tenth day. There was a statistic significant difference (P

Objective To explore the mechanism of multi-medicine drug resistance in human ovarian cancer cell line COC1/5-Fu. Methods The apoptosis and the tolerance of COC1/5-Fu cell induced by 5-Fu were analyzed by FACS. The expression of apoptosis related genes, such as p53, bcl-2, bcl-xl and bax, in COCI/5-Fu cell line were analyzed by RT-PCR. Results The COC1/5-Fu cell has some de-gree of drug resistance to 5-Fu and several other commonly used kind of chemotherapy medicine, among of which, drug resistance of 5-Fu reach 107.0 times and Paclitaxel reach 9.0 times compared with COC1. When COC1 was treated with the concentration of 5-Fu (0μmo/L, 30μmo/L or 150 μmol/L), the AI was (6.5±1.0) %, (14.0±4.0) % and (20.0±5.0) %, respectively. The rate of apoptosis increased 1.2 time and 2.1 time, compared with not treated with 5-Fu, which were significantly different (P<0.05). But when COC1/5-Fu was treated with the same concentration of 5-Fu (30 μmo/L or 150 μmoL/L), the AI was (6.7±0.7)%, (7.1±2.2)% and (6.5±2.0)%. When treated with the same concentration of 5-Fu (30 μmo/L or 150 μmol/L) , the proportion of apoptosis was significantly increased, G0/G1 phase was increased, and S and G2/ M phases were reduced in COC1 cells, but the proportion of apoptosis and cell cycle was not changed in COC1/5-Fu cells. The expression of bcl-xl , bcl-xs and bax mRNA were significantly increased and the expression of p53 and cpp32 mRNA were significantly decreased in resistant COC1/5-Fu cells , compared with COC1 cells. Conclusion wtp53 gene mutation is related with cell cycle change of ovary cancer cell and drug resistance, which is one of multi- medicine drug resistance mechanisms of COC1/5-Fu.

3-hydroxy-3-methylgulutaryl-coenzyme A (HMG-CoA) reductase inhibitors or statins are a kind of lipid-lowering agents and have been used for the prevention and treatment of cardiovascular diseases. Recent studies suggested that statins, besides lowering cholesterol, may protect vessels by enhancing the activity of endothelial nitric oxide synthase (eNOS). In the present study, we investigated if simvastatin increases eNOS activity through its phosphorylation in 293 cells (293-eNOS) with stable expression of eNOS. The results showed that incubation of 293-eNOS cells with simvastatin (10 microm/L) for 2 h significantly increased in the activity of eNOS as shown by the conversion of L-arginine to L-citrulline (2889.70+/-201.51 versus 5630.18+/-218.75 pmol/min . mg proteins) (P<0.01). Western blotting revealed that simvastatin increased phosphorylation of eNOS at 1177 (ser) and also 495 (thr) but did not affect the overall expression of eNOS or inducible NOS. Further study found that simvastatin raised phosphorylation levels of Akt and AMPK, and such effect could be antagonized by Akt inhibitor or AMPK inhibitor. These results suggest that simvastatin could stimulate the activity of eNOS via its phosphorylation by Akt and AMPK, which provides a new mechanism, other than lipid-lowering effect, for the cardiovascular protection of statins.
Cell Line ; Epithelial Cells/cytology ; Epithelial Cells/*enzymology ; Hydroxymethylglutaryl-CoA Reductase Inhibitors/*pharmacology ; Kidney/*cytology ; Nitric Oxide Synthase Type III/*metabolism ; Phosphorylation ; Simvastatin/*pharmacology

This study investigated the effects of telmisartan on insulin resistance in high-fat diet-treated rats and the possible mechanism. A total of 40 male Sprague-Dawley rats enrolled in the study were divided into 4 groups at random: ND group (n=10) and HD group (n=10), in which the rats were given a normal chow diet or a high-fat diet for 20 weeks following a one-week adaptation; ND+telmisartan (n=10) group and HD+telmisartan group (n=10), in which the rats were initially administered in the same way as the ND or HD group, and then they were orally gavaged with telmisartan (5 mg/kg daily) additionally for 5 weeks. Related inflammatory factors were measured by ELISA. Monocyte chemotactic protein 1 (MCP-1), phosphorylated JNK and IκB-α expressions in both adipose and liver were detected by Western blotting. CRP and angiotensin II receptor 1 (AT1) mRNA expressions in both adipose and liver were determined by RT-PCR. The results showed that telmisartan administration in vivo reversed insulin resistance as evidenced by a decrease in plasma fasting glucose levels, plasma fasting insulin levels and homeostasis model of assessment-insulin resistance (HOMA-IR). Furthermore, telmisartan administration significantly reduced serum CRP, TNF-α and IL-1β levels, and elevated serum IL-10 levels. It was also found to hamper the high-fat diet-induced increase in CRP mRNA, AT1 mRNA and MCP-1, and decrease in IκB-α in both adipose and liver. It was concluded that telmisartan administration in vivo may improve insulin resistance through attenuated inflammatory response pathways.

OBJECTIVETo investigate expression of the cysteinyl leukotriene receptor (CysLT1R) in hepatocellular carcinoma (HCC) tissues and determine its clinical significance.

METHODSCancerous and paraneoplastic liver tissues were collected from 30 patients with HCC and from 12 patients with liver hemangioma patients (controls). Real-time PCR and immunohistochemistry were used to evaluate the expression of CysLT1R in these tissues and assess the relationship with clinical pathological features. T-test,?Fisher's exact test and Kaplan-Meier survival analysis were used for statistical analysis.

RESULTSThe expression of CysLT1R in adjacent liver tissues (100%) was higher than that in the HCC (43.33%, P = 0.000) and normal liver tissues (41.67%, P = 0.000). The level of CysLT1R mRNA was also higher in paraneoplastic liver tissues (0.0339+/-0.0221) than in the paired HCC tissues (0.0127+/-0.0116, t = 2.911, P = 0.008) and normal liver tissues (0.0154+/-0.0123, t = -2.310, P = 0.033). There was no difference between the levels in HCC and normal liver tissues (P more than 0.05). Higher level of CysLT1R mRNA, higher level of serum alpha-fetoprotein, and higher tumor stage (III-IV) were associated with poor prognosis (respectively 4.372, P = 0.037; 24.187, P = 0.000; 8.75, P = 0.003). However, no evident relationship between the expression of CysLT1R and other clinical features was observed. Conclusions Overexpression of CysLT1R may contribute to the occurrence and progression of HCC.

Carcinoma, Hepatocellular ; diagnosis ; metabolism ; Case-Control Studies ; Disease Progression ; Humans ; Immunohistochemistry ; Kaplan-Meier Estimate ; Liver Neoplasms ; diagnosis ; metabolism ; Neoplasm Staging ; Prognosis ; RNA, Messenger ; metabolism ; Real-Time Polymerase Chain Reaction ; Receptors, Leukotriene ; metabolism ; alpha-Fetoproteins ; metabolism

. These results suggest that simvastatin could stimulate the activity of eNOS via its phosphorylation by Akt and AMPK, which provides a new mechanism, other than lipid-lowering effect, for the cardiovascular protection of statins.