Object To study the factors causing the dormancy of seed and to broaden the scope in the choice of explant for the tissue culture of the rare and near extinct plant Elaeagnus mollis Diels.Methods Dormant buds from one-year old branch collected each month from Autumn to next Spring, and seed of the very year were selected and cultured on basic media with the addition of 6-BA, NAA and GA 3 Results The germination rate of dormant bud follows the regular order of the months when it was taken. Those taken in Autumn declined every month until next Spring, and the nrevived monthly with those taken in March showing the most prosperous germination.The dormant bud can germinate into shoots. When seed was cultured, there are several factors leading to its dormancy: hardness of the mesocarp, keratinization of periderm, refractory to water permeation and the presence of certain inhibitory subtances. Seed kernal is rich in nutrition, but susceptible to bacterial infection leading to its decay. But the germination rate was well over 70% which can be cultured to give bacteria free shoots. Conclusion Both dormant buds obtained in March and bacteria free shoots germinated from naked kernal can be used as new sources for the tissue culture of E mollis .

Objective:To analyze the genotypes of HCV RNA in alcoholic and nonalcoholic HCV patients and contrast the distribution of the genotypes between those,identifying whether alcoholic patients have a higher susceptibility to certain genotype than nonalcoholic patients.Meanwhile,to detect the level of IFN-? and TNF-? for gaining the mechanism of hepatic lesion in alcoholic HCV patients.Method:114 HCV patients,according to the history of alcohol abuse,were divided into the alcohol group and nonalcohol group.The genotypes of HCV RNA in the two groups were performed by gene chip technique.Liver biopsy was practiced in 30 patients to diagnose.Ficoll gradient centrifugation and ELISA were used for extraction of PBMC and detection of IFN-?and TNF-?,respectively.Result:The genotype 1b was greater than other genotypes in the two groups,and there was no statistical significance(Alcohol group was 60% and nonalcohol group was 43.75%,P=0.22).The serum titers of HCV RNA of alcohol group were(1 1100?14 174.3)?103copies/ml and the nonalcohol group were(3 927.8?4 549.6)?103copies/ml,and there was statistical significance between the two groups(P=0.025).Among 18 alcoholic patients of 30 patients practiced liver biopsy,their liver inflammation was higher than nonalcoholic patients.The level of TNF-?of the alcohol was higher than that of nonalcohol,there was statistical significance between the two groups(P=0.048) and no statistical significance between those of IFN-?.Conclusion:Alcohol can promote the copies of HCV RNA in HCV patients and aggravate liver injury,but can't add susceptibility to certain genotypes of HCV RNA.The liver inflammation of alcoholic patients is higher than that of nonalcoholic patients.TNF-?,which builds an important effect in alcoholic HCV patients,is one of the important mechanisms in the liver injury.

Objective To explore the effect of early pelvis control training on the balance function in patients with hemiplegia. Methods 60 early stroke patients were divided into the treatment group and the control group. The patients in the treatment group accepted the early pelvis control training based on the normal early rehabilitation which was given to the control. They were evaluated with Berg Balance Scale and the Modified Bathel Index before and 2 months after treatment. Results The scores of Berg Balance Scale and the Modified Bathel Index of the treatment group improved more than that of the control group (P<0.05). Conclusion The early pelvis control training can improve the balance dysfunction after hemiplegia effectively.

Objective To investigate the prevalence of CKD in patients underging coronary angiography with suspected coronary heart disease(CHD). Methods A total of 1031patients with suspected CHD undergoing coronary angiography in Zhongda Hospital from December 2008 to October 2009 were enrolled in the study.The prevalence of CKD and associated risk factors were analyzed.GFR Wag estimated with MDRD equation.CKD was deftned as eGFR<60branches of coronary artery was considered as CHD. Results The mean age of patients were (64.37±11.02)years.There were 543 males and 488 females,including 551 patients with CHD and 134 patients with CKD(13%).Patients with CHD had a significantly higher prevalence of CKD compared with patients without CHD(18.33%vs 6.88%,P<0.01).With the increasing number of stenosis coronary vessels(n=0,1,2,3),eGFR was declined[(84.25±19.00),(81.61±23.92),increased(0.42%,0.82%,1.96%,3.25%,P=0.006),and the prevalence of CKD increased (6.88%,13.11%,21.57%,23.38%,P<0.01).Logistic regression analysis indicated that increasing age(OR 1.094,95%C/1.068 to 1.120),increasing number of stenosis coronary vessels(OR 1.288,95%CI 1.074 to 1.543).hypertension(OR 1.974,95%CI 1.082 to 3.603),cardiac systolic insufficiency(OR 3.183,95%CI 1.696 to 5.972),and hyperuricemia(OR 5.366,95%CI 3.224 to 8.9311 were risk factors for CKD. Conclusions The prevalence of CKD in patients with CHD diagnosed by coronary angiography is quite high.Aging,elevated number of stenosis coronary vessels,hypertension,cardiac systolic insufficiency and hyperuricemia are important risk factors for angiographic patients with CKD.

Objective To establish a sandwich enzyme-linked immunosorbent (ELISA) method for early diagnosis of hepatitis C. Methods Immunization of Balb/c mice with hepatitis C core antigen were prepared by genetic engineering. Mouse monoclonal antibodies (McAb) to anti-HCV-core Ag were obtained. A sandwich ELISA kit for detecting HCV-core Ag was developed by using four strains of anti-HCV-core Ag McAb. One hundred and twenty five serum specimens with increased ALT but negative for anti-HCV, anti-HIV, and anti-Tp tests were tested. Results Nine of the 125 specimens were positive for HCV-core Ag. Conclusion The double sandwich ELISA kits for detecting HCV-core Ag may be useful for the early diagnosis of hepatitis C .

Objective To construct prokaryotic expression vectors for glutamate dehydrogenase(GDH)of Clostridium difficile(CD),and express recombinant GDH in Escherichia coli,and identify its antigenicityed.Methods The entire gene of GDH was cloned from ATCC43255 genome DNA.The recombinant antigens were expressed in E.coli with IPTG induction and purified by Ni-NATBeads.The antigenicity was detected using CD Qick Chek Complete dual-antigen EIA. Results Prokaryotic expression vectors of CD GDH were constructed successfully.The antigen could be identified by specific anti-GDH antibodies.Conclusion The GDH antigen can be used to prepare corresponding antibodies,which facilitate the development of immunoassay for CD GDH.

Objective To design a complex hepatitis C vires(HCV)=E1 antigen,and to search its application in HCV vaccine and diagnosis test.Methods Through consulting the database and widely comparison of sequences from HCV E1 of different genotype,the representative immunodominant epitope sequences were selected from all the six genotypes.Their genes were deduced according to the preference codon in E.coli and three fragments were designed to contain the six epitopes.They were chemically synthesized and cloned into pBVIL1 vector separately.The cloned fragments were conjugated together profiting from pBVIL1's property,and then a doubled pan-DR helper T cell epitopes(PADRE)gene was inserted to form a complex expression plasmid.The transformed E.coli cells with this plasmid were cultured and induced to express recombinant protein and the antigenic activity of the product was tested.Results An expressing plasmid containing 8 epitopes from HCV genotype 1a,1b,2a,3a,4a,6a and a doubled PADRE sequences was constructed successfully and the engineering E.coli transformed with this plasmid highly expressed after inducing culture at 42℃ in an inclusion manner.The immunological activity of the purified recombinsnt multi-epitope antigen shows that:(1)It can react with a great part of sera from HCV positive patients by indirect ELISA.(2)It can induce notable specific humoral immunity in injected mice.Conclusion The novel constructed expressing plasmid and its product may be useful in study of a newly HCV vaccine as well as an antigen for HCV immunoassays.

The gelatinization process of the starch is replaced by unpolluted steam-pretreatment on the base of the Radix Puerariae rich in fiber and isoflavones. The production of ethanol and isoflavones by simultaneous saccharification and solid state fermentation (SSF) of steam-pretreatment Radix Puerariae is presented. The optimal technological conditions were obtained: Radix Puerariae being steam-pretreated at a saturated vapor pressure of 0.8 MPa for 3.5 min, glucoamylase(65 u/g), cellulase(1.5 u/g), 0.1%(NH4)2SO4, 0.1%KH2PO4 and activated yeasts being added in, and fermentation at 35-37 degrees C for 60 h. Under these conditions, the yield of ethanol and isoflavones from 100 g Radix Pureriae (dry basis) were 27.47 g and 4.43 g, respectively, the starch utilization rate was 95%. In comparison with the traditional fermentation technology, the simultaneous saccharification and SSF of steam-pretreatment Radix Puerariae is clean and energy-saving. It provides new way of the production of ethanol from the non-food starch material, and worthwhile to be explored and implemented in industry.
Cellulase ; metabolism ; Ethanol ; metabolism ; Fermentation ; physiology ; Glucan 1,4-alpha-Glucosidase ; metabolism ; Isoflavones ; biosynthesis ; Pueraria ; metabolism ; Steam ; Yeasts ; metabolism

AIM: To investigate the effect of huayu xiaoliu fang, a Chinese medicine, on the cell cycle of human lung carcinoma cell line by serologic pharmacological method. METHODS: PGLH7 cells were incubated with rabbit serum containing huayu xiaoliu fang at different doses obtained by serologic pharmacological method. MTT assay was used to calculate the proliferation inhibition rate. The target cells were harvested to analyze the cell cycles by flow cytometry. RESULTS: The Chinese medicine-containing serum inhibited the growth of PGLH7 cells significantly. There was remarkable difference in the proliferation inhibition rate between 10% (high dose) Chinese medicine-containing serum and the control serum (P