Objective To investigate the clinical effect of thymosin combined with liver gland peptide drugs in the treatment of primary liver cancer.Methods 138 cases of primary hepatocellular carcinoma (HCC) were seleted as the research subjects,and they were divided into observation group (n=69) and control group (n=69) according to the number table method.The control group was treated with simple hepatoprotective drugs,the observation group was treated with thymosin on the basis of the control group.The clinical curative effect of the two groups,quality of life improvements were observed.Results In the observation group,the total effective rate was 81.16%,which in the control group was 43.47%,the difference was statistically significant between the two groups (χ2=8.362,P<0.05).The rate of life quality improvement of the observation group was 60.86%,which of the control group was 23.18%,there was statistically significant difference between the two groups (χ2=9.726,P<0.05).Conclusion The primary HCC patients treaed with thymosin combined with hepatoprotective drugs can effectively improve the patients' life quality,with good clinical curative effect,and it is worth clinical promotion and application.

By furthering microbiology experiment reform,enforcing "high starting point,high efficiency,high quality"teaching,adjusting teaching contents,reforming teaching methods,establishing and promoting testing system,a new microbiology experiment curriculum system is to be established with the basic knowledge and skills as basic contents and cultivating creative ability as central contents.This will motivate the students' interests for microbiology experiment,which helps them to improve their ability of thinking independently and creatively as well as their practicing ability.

Objective To express mouse IFN-λ2 stably and study its biological activity. Methods Full-length of mIFN-λ2 cDNA was obtained by using RT-PCR from cells of mouse spleen stimulated by ve-sicular stomatitis virus(VSV) and then subcloned to eukaryotic expressing vector PCAGG-EGFP. The recom-binant was transfected into CHO cells. VSV * GFP-B16 system was used to measure the antivirus activity. The constructed cell line MDBK-Mxp-Luc was used to study the character of Mx1 protein induced by the mIFN-λ2. Results The recombinant pMD18-T-mIFN-λ2 was digested by two kinds of enzyme, Sac I and Xho I, to produce the fragment was of 582 bp, and of which the sequence analysis of sequence shows it was entirely consistent with the nucleotide sequences reported in GenBank. PCAGG-EGFP-mIFN-λ2 eukaryotic expressing vector was constructed successfully and expressed stably in CHO cells, and the mRNA of mIFN-λ2 was verified expressing in CHO-PCAGG-EGFP-mIFN-λ2 cell line by RT-PCR. The antivirus activity of in the supernatant secreted by the CHO-PCAGG-EGFP-mIFN-λ2 cell line was 10~4 AU/ml. The mIFN-λ2 pro-tein can could induce the expression of the antivirus protein Mx1, and the expression of Mx1 protein induced by mIFN-λ2 enhanced with time going, 9 to 12 hours achieved the peak, 24 hours vanished. Conclusion Gene cloning of mIFN-λ2 was successful. The eukaryotic expressing vector of mIFN-λ2 was constructed suc-cessfully and expressed stably in CHO cells, and its product has obvious antivirus activity in vitro. And the antivirus activity of the product was closely correlated with inducing expression of antivirus protein Mx1.

ObjectiveTo aralyze The prevalence of premature children, mortality and complications to related factors, discussed measures to reduce the premature children mortality. MethodsFrom January 2003 to December 2006 and January 2007 January 2010 12 after 8 years in our hospital obstetric preterm children born in 1110 into the first 4 years of group and group 4 years after the two groups. Each group of children on the incidence of premature causes of death, complications were retrospectively analyzed. Results Preterm birth rate of first 4 years 2.85％ to 3.67％ after 4 years,the difference was statistically significant( P ＜0.05 ). Between the two groups the proportion of different age groups and the main types of diseases,no significant difference(P＞ 0.05). By comparison,the mortality of premature children dropped from the 12.06％ to 2.36％, a significant difference before and after ( P ＜ 0.01 ).ConclusionThe prevention and treatment of premature children was still very trduous task, particularly in preventing complications of preterm children was very important to prevent the occurrence of lung disease.

Objective To observe the effect of electroacupuncture (EA) plus oxygenmedicine (OM) on the expression of Bcl-2 and Bax in the hippocampal CA 1 area in cerebral ischemia/reperfusion injury (CI/RI) rats. Methods Thirty SD rats were randomized into sham-operation,model,EA,OM,EA+OM groups (n=6/group). CI/RI model was established by using modified Pulsinelli 4 vessel occlusion and reperfusion. EA (100 Hz,3.5 mA) was applied to "Baihui" (GV 20) and "Zusanli" (ST 36) 30 min,once daily for 4 days. Rats of OM and EA+OM groups were put into a box filled with oxygen and atomized herbal medicines containing Bingpian (Borneolum),Shexiang (Moschus),Huangjing (Rhizoma Polygonati),Shouwu (Radix Polygoni Multiflori),etc. for 30 min,once daily for 4 days. Bcl-2 and Bax expression of the hippocampal CA 1 area was detected by immunohistochemistry. Results Compared with sham group,the numbers of Bcl-2 immunoreaction (IR) and Bax IR positive cells,and the immunoactivity of Bcl-2 IR and Bax IR positive products in the hippocampal CA 1 area were increased significantly in model group (P0.05). Conclusion EA and OM and EA+OM can effectively regulate the expression of Bcl-2 and Bax in the hippocampal CA 1 area in CI/RI rats,and the effects of EA+OM are significantly superior to those of simple EA and simple OM,which may contribute to their effect in improving cerebral ischemia.

To achieve higher level expression of Interferon α2b (IFN-α2b) in methylotrophic yeast (Pichia pastoris), a cDNA fragment coding for the mature IFN-α2b was designed and synthesized based on the synonymous codon bias of P. pastoris and optimized G+C content. The synthetic IFN-α2b was inserted into the secreted expression vector pPICZαA, and then integrated into P. pastoris GS115 genome by electroporation. Multi-copy integrants in the Mut+ recombinant P. pastoris strain were screened by high concentrations of Zeocin. 120 hours culturing allowed expression of the IFN-α2b transformant up to 810 mg/L as detected by SDS-PAGE and quantitative methods. In addition, Western blot analysis showed that the recombinant proteins had immunogenicity. The significant antiviral activity of the recombinant IFN-α2b protein was verified by WISH/ VSV system, which was 3.3×105 IU/mL.

Objective To explore the differentiation potential of bone marrow stromal cells (BMSC) to enteric neuron in vitro and to seek proper induction methods.Methods BMSC were harves- ted from male rats and cultured in DMEM supplemented with 20% fetal bovine serum,and characterized by flow cytometry.At passage 6,BMSC were pre-induced by basic fibroblast growth factor (bFGF,10 ng/ml) for 24 h,then induced in two groups:glial cell-line derived neurotrophic factor (GDNF) group, 10 ng/ml GDNF in fetal gut condition medium (FGCM) for 10 d.Vitamin A acid (VA) group,VA, zinc in FGCM for 10 d.The expressions of neuronal markers,neural specific enolase (NSE) and neu- rofilament (NF),glial cell marker,glial fibrillary acidic protein (GFAP),enteric neuronal marker,pro- tein gene production 9.5(PGP9.5),nitric oxide synthase (nNOS),enteric neural transmitter,vasoactive intestinal peptide (VIP) were detected by fluorescent immunohistochemistry method.Results The cul- tured BMSC were CD90 positive and CD45 negative on flow cytometry.After 10 d of induction,a certain number of cells adopted neuron-like morphological changes and showed the expressions of NSE,NF, PGPg.5,nNOS and VIP without the expression of GFAP by fluorescent immunohistoehemistry method in both groups.But in GDNF group,the positive rate of NF,PGP9.5,nNOS and VIP was significantly higher than that in VA group (75.6%?8.4% vs 48.5?7.5%;57.7%?6.5% vs 35.7%?7.2% 46.6%?5.4% vs 30.5%?6.6%;72.3%?6.7% vs 40.4%?7.4%;P＜0.01).Conclusion BMSC can be induced to differentate into enteric neuron in vitro by different methods.GDNF with FGCM can induce higher rate of enteric neuron like cells compared with VA etc.

OBJECTIVETo investigate the impact of the Artemisia annua plant-derived drug, artesunate, on proliferation of primary rat hepatic stellate cells (HSCs), and to analyze the underlying molecular mechanisms of its anti-fibrogenic effects involving the inhibition of transforming growth factor-beta 1 (TGF-b1) expression and secretion in liver.

METHODIsolated, cultured, and activated primary rat HSCs were divided into sixteen groups, including one untreated control group and fifteen artesunate-treated experimental groups with 125, 150, 175, 200 or 225 mumol/L for 24, 48 or 72 hours. The rate of cellular proliferation was measured using the 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay. TGF-b1 mRNA expression was evaluated by reverse transcription-polymerase chain reaction and protein expression was evaluated by Western blotting. Enzyme-linked immunosorbent assay was used to evaluate secreted levels of TGF-b1 protein.

RESULTSArtesunate significantly inhibited proliferation of cultured HSCs in a dose- and time-dependent manner (all, P less than 0.01). After 24 hours of exposure, the inhibition ratios of the various artesunate concentrations were: 6.06%+/-1.44% (125 mumol/L), 21.47%+/-5.57% (150 mumol/L), 42.00%+/-7.36% (175 mumol/L), 67.12%+/-4.55% (200 mumol/L), and 79.83%+/-3.67% (225 mumol/L). Artesunate significantly inhibited the TGF-b1 mRNA expression in HSCs, and the higher the drug concentration, the higher the degree of inhibition (all, P less than 0.01). In addition, artesunate significantly inhibited the expression of intracellular and secreted TGF-b1 protein (all, P less than 0.01). In response to artesunate (mumol/L concentrations), the TGF-b1 levels were (164.24+/-6.88) pg/ml (0μmol/L), (102.68+/-4.45) pg/ml (150μmol/L), (86.54+/-5.56) pg/ml (175μmol/L), and (56.55+/-5.66) pg/ml (200μmol/L).

CONCLUSIONArtesunate exerts anti-fibrogenic effects on HSCs in vitro, possibly by reducing the expression, translation and secretion of TGF-b1.

Animals ; Artemisinins ; pharmacology ; Cells, Cultured ; Hepatic Stellate Cells ; drug effects ; secretion ; Rats ; Rats, Wistar ; Transforming Growth Factor beta1 ; metabolism

Objective To study the influence factors for HBV reinfection following liver transplantation.Methods 92 cases of liver transplantation were enrolled for open non-randomized clinical study.Given conventional OLTx and immunosuppressive agents and antibiotics to prevent infection etc,patients are divided into lamivudine alone and HBIG combined with lamivudine group.Observation of postoperative liver function,HBV serum markers,HBV DNA in PBMC,YMDD and HBV S gene mutation,liver tissue IH etc.Results In 92 cases of liver transplantation with HBV-related liver disease,hepatitis B recurrence rate was 4.35%(4/92).In lamivudine group the HBV infection rate was 35%(7/20);In combined therapy group the HBV infection rate was 6.94%(5/72).With preoperatively negative HBV DNA negative the hepatitis B recurrence rate was 0;With preoperatively positive HBV DNA,the postoperative HBV infection rate was 17.39%(12/69),which had cases with S gene or YMDD mutation.In patients with negative HBV DNA before and after operation,the HBV re-infection rate was 11.11%(1/9);In 5 cases with preoperative HBV DNA positive,the HBV infection rate was 4/5 after LTx;before and after operation HBV DNA are positive,the HBV infection rate was 100%(3/3);the preoperative HBV DNA positive and postoperative HBV DNA negative,the HBV infection rate was 50%(1/2).Conclusion To prevent HBV infection after LTx,lamivudine group easily leads to HBV re-infection than HBIG combined with lamivudine group.Preoperative serum HBV DNA positive,preoperative and postoperative HBV YMDD and S gene mutation are the primary factors affecting the HBV re-infection after operation.HBV DNA positive in PBMC is the source of HBV re-infection,but also the factor of recurrence of hepatitis B.

BACKGROUND:Recently,acellular dermal matrix allograft has been widely used in the repair of oral mucosal defects.But little information is about the heterogeneous acellular dermal matrix (HADM) patch for repair of oral mucosal defects.OBJECTIVE:To investigate the efficacy and biosafety of HADM in the repair of oral mucosal defects.METHODS:In total 71 patients with oral benign or malignant tumors who had oral mucosal or soft tissue defects following tumorectomy were included in this study.These patients comprised 37 males and 34 females,and were averaged 45 years (range,20-70 years old).Of them,42 suffered from benign tumors and 29 from malignant tumors.HADM patches were used for repair of oral mucosal defects.The survival,color,and texture of HADM patches were observed.Shrinkage rate of HADM patches was compared between regions without supports from hard tissues (cheeks,tongue,and mouth floor) and with supports from hard tissues (gingiva,hard palate).RESULTS AND CONCLUSION:All 71 HADM completely survived.No necrosis and infection occurred.At 2 weeks after transplantation,(98.20±5.20) % of patch area survived.At 3 months after transplantation,patches showed similar color to surrounding oral mucosa and most patients had sense of tension to different extents.At 6 months after transplantation,cell creeping substitution and vasculadzation were successfully accomplished in the region of patch transplantation.Patches grew stably,with smooth pink appearance and good elasticity,and no further shdnkage.Patients felt normal.HADM patch shrank primarily at 2 weeks-1 month after transplantation,and tended to be stable at 3 months.There was no significant difference in tissue morphology between surgical region and normal tissue.The HADM shdnkage rate was significantly higher in regions without supports from hard tissues than regions with supports from hard tissues.These findings indicate that HADM patches have advantages in repair of oral mucosal defects including good histocompatibility,wide source,simple manipulation,and able to cover the wound surface in the early state,promote wound surface healing,and reduce scar formation,and can be used as an ideal matedal for repair of oral mucosal defects.