Aim:to detect how Rb-deficiency will affect responses to TGF-? induced cell cycle arrest of hepatocyte.Methods:primary hepatocytes were isolated and cultured,and Rb-specific adenovirus siRNA was transformed into cells.Then TGF-? was added daily and cell growth and cell cycles variations.Western blot and FQ-RT-PCR were adopted to detect pRb,E2F,c-MYC and p16 expression changes.Results:Primary hepatocytes were isolated and infected with Rb-specific siRNA adenovirus.In control cells,treatment with TGF-? prevented cells to enter S phase via decreased cMYC activity,activation of P16INK4A activity.In Rb-null hepatocytes,E2F and cMYC activity decreased slightly but P16INK4A was not activated and the great majority of cells continued cycling.Rb is therefore central to TGF?-induced cell cycle arrest in hepatocytes.Conclusion:The present results show that otherwise genetically normal hepatocytes with disabled Rb genes respond less well to the antiproliferative effects of TGF-?.

Objective Eucheuma is rich in nutrients and can be an important raw material of food after processed. This study was designed to establish a feasible method of purifying polypeptides from eucheuma and investigate their activity against platelet aggre?gation and bacterial growth. Methods We extracted peptides from eucheuma with acidic solution, detected the effects of different doses of small molecular polypeptide ( 0, 5, 10, 20, and 40μg/mL) on the growth of Escherichia coli ( D1314) and Staphylococcus aureus (s.agr+, RN4220) using the method of turbidity, and analyzed the anti?platelet aggregation activity of the peptides with a whole blood aggregometer. Results The rates of peptides extracted from 50, 100,150, and 200 g of eucheuma were 0.382%, 0.405%, 0.389%, and 0.389%, respectively. The purified sample exhibited a single band on SDS?PAGE. The relative molecular weight of the peptides was about 3kD. The extracted peptides inhibited the growth of Escherichia coli, Staphylococcus aureus, and thrombin?induced platelet aggregation in a dose?dependent manner, with inhibition rates of 44.71%, 51.86%, and 75.00%, respectively. Conclusion The present method can be used to successfully purify low?molecular?weight peptides from eucheuma and effectively inhibit platelet aggre?gation and bacterial growth. The peptides extracted is a potential anti?platelet aggregation agent.

Objectives To investigate the changes and features of drug resistance in Haemophilus inlfuenzae (Hi) isolated from children with infection diseases in Suzhou. Methods One thousand two hundred and twenty-two Hi strains isolated from clinical specimens were collected from January 2011 to June 2012. Antimicrobial susceptibility was tested by Kirby-Bauer me-thod, andβ-lactamase was analyzed by chromogenic nitroceifn method. Then strains were divided into four groups according to their speciifc resistance patterns:①β-lactamase positive strains (except for cefuroxime-resistant strains),②β-lactamase negative ampicillin-resistant (BLNAR) strains,③cefuroxime-resistant strains, and④other resistant strains. Results The resistance of iso-lated Hi from children in Suzhou area to ampicillin, co-trimoxazole, cefaclor, tetracycline, chloramphenicol, azithromycin, cefu-roxime, ampicillin/sulbactam was 32.7%, 76.7%, 27.4%, 14.3%, 10.2%, 8.4%, 6.9%and 4.3%, respectively, and the detection rate in four groups was 29.0%, 2.6%, 6.9%, 41.2%, respectively. Multi-resistant strains were mainly distributed in Group①and Group②, accounting for 67.5%and 81.3%of each group. Azithromycin, tetracycline and chloramphenicol showed high antimicrobial activity against BLNAR and cefuroxime-resistant Hi strains. The positive rate of resistance toβ-lactamase was 30.8%. The rates of resistance to cefuroxime, cefaclor, ampicillin/sulbactam and cotrimoxazole inβ-lactamase-positive ampicillin-resistant strains are signiifcantly different from those inβ-lactamase-negative ampicillin-resistant strains. Conclusions The resistance feature of Hi isolated from children in Suzhou shows signiifcant changes, including new appearance of cefuroxime-resistant strain, a rapid increase in resistance to azithromycin, and a large proportion of multidrug-resistant strains. The rapid increase in BLNAR and the emergence of cefuroxime-resistant strains have become the new resistance pattern of Hi in this area.

BACKGROUNDUroacitides is a group of cell differentiation inducers, which is purified from fresh human urine. Preclinical studies of Uroacitides have showed that cancer cells could be induced to differentiate, and the growth of cancer cells could be inhibited by Uroacitides. The aim of this study is to compare the efficacy and toxicity between Uroacitides combined with NP regimen and NP alone in treatment of advanced non-small cell lung cancer (NSCLC).

METHODSForty-two cases of advanced NSCLC were randomized into Uroacitides+NP and NP groups. NP group: NVB 25mg/m² on days 1 and 8, DDP 75mg/m² on day 1. Uroacitides combined with NP group: Uroacitides of 300mL was given through subclavian catheter daily for 7 days prior to the NP chemotherapy, then concurrently with NP regimen for 2 cycles, except the days of administration of chemotherapy.

RESULTSIn the Uroacitides+NP group, the overall response rate was 44.4%, and 20.0% in the NP group (P > 0.05). The median survival time was 9 months in the Uroacitides+NP group and 6 months in the NP group (P=0.0287). The main toxicities were myelosuppression, gastrointestinal reaction and alopecia, and there was no significant difference in incidences of toxicities between the two groups (P > 0.05).

CONCLUSIONSUroacitides combined with NP regimen shows a good curative effect and low toxicity, and may significantly prolong the median survival time for advanced NSCLC.

OBJECTIVE：To study the effects of paeonol combined with cisplatin on the proliferation and apoptosis of human osteosarcoma cell MG- 63 and its possible mechanism. METHODS ：MG-63 cells in logarithmic growth phase were divided into blank control group ，cisplatin group （4 µmol/L），paeonol group （50 mg/L），and low ，medium，high concentration combined groups （50，100，200 mg/L paeonol+ 4 µmol/L cisplatin ）. CCK- 8 method was used to detect the cell proliferation rate at 24，48，and 72 hours of treatment. Annexin Ⅴ-FITC/PI double staining method was used to detect the cell apoptosis rate at 24 hours of treatment. The relative expression of PI 3KCA，Akt，mTOR，P-gp and PTEN mRNA in cells were detected by qRT-PCR. RESULTS ：Compared with blank control group ，the cell proliferation rate at each time point ，and the relative expression of PI 3KCA，Akt and mTOR mRNA in cells were significantly reduced and the apoptosis rate and the relative expression of P-gp and PTEN mRNA in the cells were increased significantly （P＜0.05 or P＜0.01）. Compared with cisplatin group and paeonol group ，cell proliferation rate at each time point and the relative expression of PI 3KCA，Akt and mTOR mRNA in cells were decreased significantly in the high concentration combination group ，while the relative expression of P-gp and PTEN mRNA in the cells were significantly increased （P＜0.01）；there were statistical significance in some of the above indicators in the medium and low concentration combination groups（P＜0.05 or P＜0.01）. CONCLUSIONS ：The combination of paeonol and cisplatin could inhibit the proliferation of MG- 63 cells and promote their apoptosis ，which may be related to the down-regulation of PI 3KCA，Akt and mTOR mRNA expression ，and the up-regulation of P-gp and PTEN mRNA expression.

BACKGROUNDTo evaluate the efficacy, side-effects, median survival duration and survival rate of vinorelbine (NVB) combined with cisplatin (DDP) in the treatment of non-small cell lung cancer (NSCLC).

METHODSA total of 220 patients with inoperable NSCLC received NVB and DDP combined chemotherapy: NVB 25-30 mg/(m²*d) on days 1 and 5 (or 8), DDP 60-80 mg/(m²*d) on day 2. The schedule was repeated every 28 days. The efficacy and side-effects were analysed and followed-up after at least two cycles of chemotherapy.

RESULTSThe overall response rate (CR+PR) was 30.9% (68/220). The response rate was 31.3% (51/163) in initial treatment group, and 29.8% (17/57) in retreatment group. The median survival duration was 8.3 months. The 1-, 2- and 3-year survival rates were 39.23%, 19.31% and 6.32%, respectively. The main side-effects were myelosuppression and digestive tract reactions.

CONCLUSIONSVinorelbine plus cisplatin is an effective and well-tolerated regimen for non small cell lung cancer and myelosuppression is its dose-limiting toxicity.

Objective: To explore the persistent viral response rate (SVR) in patients with refractory chronic hepatitis C after interferon (IFN) (peginterferon 360 μg qw) and ribavirin (PR) therapy failure. The SVR of patients with refractory chronic hepatitis C was improved by PR combined with direct antiviral agents (DAA) and proper extension of the course of therapy was applied. Methods: Seventeen cases of refractory chronic hepatitis C after IFN(peginterferon 360 μg qw) and ribavirin therapy failure were given PR combined with DAA treatment. The side effects were observed and corresponding adjustments were made on drug dosage, and SVR was recorded. Results: The 17 cases completed the whole course of treatment with PR combined with DAA for 24 weeks. All the 17 patients obtained rapid viralogical response (RVR) and SVR. After treatment, the SVR rate was 100% in patients including those with virologic relapse, retreated or previously non-responsive patients with refractory chronic hepatitis C. The adverse reaction of PR combined with DAA 24 weeks was generally mild. Conclusions The use of PR combined with DAA re-treatment in patients with refractory chronic hepatitis C can achieve SVR and shorten the treatment time. PR combined with DAA re-therapy is one of effective treatments to improve the rate of sustained viral response in patients with refractory chronic hepatitis C.

BACKGROUNDEndostar™ (rh-endostatin, YH-16) is a new recombinant human endostatin developed by Medgenn Bioengineering Co. Ltd., Yantai, Shandong, P.R.China. Pre-clinical study indicated that YH-16 could inhibit tumor endothelial cell proliferation, angiogenesis and tumor growth. Phase I and phase II studies revealed that YH-16 was effective as single agent with good tolerance in clinical use.The current study was to compare the response rate , median ti me to progression (TTP) ,clinical benefit andsafety in patients with advanced non-small cell lung cancer ( NSCLC) , who were treated with YH-16 plus vi-norelbine and cisplatin (NP) or placebo plus NP.

METHODSFour hundred and ninety-three histologically or cy-tologically confirmed stage IIIB and IV NSCLC patients , withlife expectancy > 3 months and ECOG perform-ance status 0-2 , were enrolledin a randomized ,double-blind ,placebo-controlled , multicenter trial ,either trialgroup : NP plus YH-16 (vinorelbine 25 mg/m² on day 1 and day 5 ,cisplatin 30mg/m² on days 2 to 4 , YH-167.5mg/m² on days 1 to 14) or control group : NP plus placebo (vinorelbine 25 mg/m² on day 1 and day 5 ,cis-platin 30 mg/m² on days 2 to 4 ,0.9% sodium-chloride 3 .75 ml on days 1 to 14) every 3 weeks for 2-6 cycles .The trial endpoints included response rate ,clinical benefit rate ,time to progression,quality of life and safety .

RESULTSOf 486 assessable patients , overall response rate was 35.4% in trial group and 19.5% in controlgroup (P=0 .0003) . The median TTP was 6 .3 months and 3 .6 months for trial group and control group respectively (P < 0 .001) . The clinical benefit rate was 73 .3 %in trial group and 64.0% in control group (P=0 .035) .In untreated patients of trial group and control group ,the response rate was 40 .0% and 23.9%(P=0 .003) ,the clinical benefit rate was 76 .5 % and 65 .0 % (P=0 .023) ,the median TTP was 6 .6 and 3 .7months (P=0 .0000) ,respectively .In pretreated patients of trial group and control group ,the response ratewas 23.9% and 8.5%(P=0 .034) ,the clinical benefit rate was 65.2% and 61.7%(P=0 .68) ,the median TTP was 5 .7 and 3 .2 months (P=0 .0002) ,respectively . The relief rate of clinical symptoms in trial groupwas higher than that of those in control group ,but no significance existed (P > 0 .05) . The score of quality oflife in trial group was significantly higher than that in control group (P=0 .0155) after treatment . There were no significant differences in incidence of hematologic and non-hematologic toxicity , moderate and severe sideeffects betweentrial group and control group .

CONCLUSIONSThe addition of YH-16 to NP regimen results in significantly and clinically meaningful improvement in response rate , median time to tumor progression,and clinical benefit rate compared with NP alone in advanced NSCLC patients . YH-16 in combination with chemotherapy shows a synergic activity and a favorable toxic profile in advanced cancer patients .

Objective: To investigate the differences in function of plasmacytoid dendritic cells (pDC) and CD4⁺ T helper cells (CD4⁺ Th cells) between acute hepatitis B (AHB) and chronic hepatitis B (CHB). Methods: In this study, patients with AHB and those with CHB in immune active (IA) phase were enrolled. The frequencies of pDC, CD86⁺ pDC, CD4⁺ T cells and their subsets, surface functional molecules were detected respectively among patients with chronic HBV infection in IA phase, patients with AHB, those recovered from AHB. Meanwhile, their correlations with ALT, HBV DNA and HBV markers were analyzed. Results: The ALT level in AHB was significantly higher than that in IA, and inflammation was more obvious in AHB. Between IA and AHB, CD86⁺ pDC frequency and the mean fluorescence intensity of functional molecule CD86 (CD86MFI) were higher in IA than those in AHB, but the frequency of CD4⁺ T cells in AHB was higher than that in IA. For patients who got over AHB, the frequency of CD86⁺ pDC increased; Th1 were on the rise, while the frequencies of CD4⁺ T and Th2 decreased after the recovery of AHB, and Th2 / Th1 ratio decreased..In AHB, HBVDNA loads were positively correlated with ALT levels and Th2 frequencies. Conclusions In CHB immune active phase, CD86⁺ pDC with stimulating function played an important role, but the cellular immune response of CD4⁺ T cells decreased. In AHB inflammatory stage, CD4⁺ T cells played a strong cellular immune response, which result ed in viral clearance. Th2 cells regulation of CD4⁺ T cells played a dominant role, which was involved in the inflammatory response, and the cytotoxic role of Th1 cells during the recovery period was dominant, playing a strong cellular immune response, then the virus were completely eliminated.

Objective: To investigate the differences in frequency and function of natural killer cells (NK) between chronic hepatitis B (CHB) and acute hepatitis B (AHB). Methods: Patients with AHB and those with CHB in immune active (IA) phase were enrolled. The frequencies of NK, CD56^dimNK, CD56^brightNK and the expression of functional molecules IFNAR2 and NKp46 on the surface of NK cells were detected respectively among patients with CHB in IA phase, patients with AHB, and those recovered from AHB. At the same time, their correlations with ALT, HBV DNA and HBV markers were analyzed. Results: Between IA and AHB, the frequencies of NK cells and NKp46^dim NK cells in AHB cases were significantly lower than those in IA cases, but the frequency of NKp46^high NK cells in AHB was higher than that in IA. For patients who recovered from AHB, the frequency of NK cells and NKp46^dim NK cells increased; the varied ranges of frequencies of CD56^dimNK, IFNAR2⁺ NK and NKp46⁺ NK cells were on the rise, while the frequency of NKp46^high NK cells decreased after the recovery from AHB, and the varied ranges of CD56^brightNK and IFNAR2MFI, NKp46MFI decreased. In AHB, HBVDNA loads were positively correlated with ALT levels. Before and after the recovery of AHB: ΔHBV DNA and ΔALT, Δ NK/LY (%) were positively correlated; ΔALT and ΔNKp46^highNK/NK(%), ΔNKp46MFI, ΔIFNAR2MFI were positively correlated. Conclusions In CHB immune active phase, the activity of peripheral blood NK cells was too weak to remove the virus, but NK cells play an important role in eliminating the viruses and mediating liver tissue inflammation in AHB.