2.Study of Breeding Saccharomyces cerevisiae with Improved Temperature and Ethanol Tolerance by Genome Shuffling
Hao WANG ; Hang WANG ; Chun MENG ; Yang-Hao GUO ;
Microbiology 1992;0(04):-
By UV induced mutagenesis of protoplasts of Saccharomyces cerevisiae strain f4, f5 and f6, and screening on plates containing different concertration of ethanol at different temperature, we obtained improved strains, such as f4.2, f5.1, f6.2, f4.5. By using a DES to deal with all the improved strains, We obtained two mutants, f5.1.1 and f4.2.1,which have improved ethanol torlerance. We made use of genome shuffling to generate improved strains in this work. We shuffled twice these improved strains by protoplast fusion and finally obtained strains with higher temperature and ethanol tolerance. we also identified shuffled strains that produced more ethanol by shake-flask experments. At the 35℃, the ethanol yield of R24 strain got to 12.93% (W/V), and were almost 5% higher than that of stain f4.
4.Silencing of miR-21 influences the function of colon cancer cell line HT-29 and the expression of PDCD4
Yupeng REN ; Chun SONG ; Hao ZHANG
China Oncology 2015;(1):6-12
Background and purpose: PDCD4 may be inhibited by miR-21 to regulate the malignant behaviors of colon cancer such as invasion and migration. This study aimed to explore the function of colon cancer HT-29 cell lines by downregulating miR-21 expression and discuss the mechanisms and relationship between miR-21 and PDCD4 in colon cancer malignant behaviors. Methods:simiR-21 was transfected into colon cancer cell line HT-29 to downregulate the expression of miR-21. Proliferation, apoptosis, migration and invasion were detected by MTT, flow cytometry and Transwell assay after transfection. PDCD4 expression was detected by Western blot and qRT-PCR. Results:The qRT-PCR analysis result proved that the transfection efifciency was 60%-65%. MTT analysis result showed that the proliferations of HT-29 cells were inhibited after the transfection of miR-21 for 72, 96, 120 h (t=1.276, P<0.05;t=3.276, P<0.01;t=4.523, P<0.01). Comparing with si-negative control and miR-21 groups, lfow cytometry result showed that the apoptosis rate was increased after miR-21 expression downregulated (t=2.132, P<0.05;t=3.524, P<0.05). Transwell assay result showed that migration (t=2.423, P<0.05; t=3.153, P<0.05) and invasion(t=3.245, P<0.05; t=5.236, P<0.05) were inhibited;Western blot result showed that PDCD4 expression was up-regulated at protein level(t=2.342, P<0.05;t=4.215, P<0.05);qRT-PCR result showed that PDCD4 expression was up-regulated at mRNA level(t=2.261, P<0.05; t=3.492, P<0.05). Conclusion: The proliferation, migration and invasion are the inhibited, and apoptosis is attenuated after miR-21 downregulated by simiR-21 transfection, PDCD4 expression is up-regulated. miR-21 may enhance the malignant behavior of cancer cells by downregulating the PDCD4 expression, miR-21 might be a target gene for colon cancer therapy.
5.Clinical analysis of 109 cases of isolated fetal pyelectasis and pregnancy outcome
Hao ZHU ; Chun SHEN ; Xiaotian LI
Chinese Journal of Perinatal Medicine 2015;18(9):683-686
Objective To investigate the prognosis of isolated fetal pyelectasis detected by ultrasound.Methods A total of 109 cases of isolated fetal pyelectasis (renal pelvis anteroposterior diameter ≥ 5 mm by ultrasound screening at any gestational age without structural or chromosome abnormalities) detected by prenatal ultrasound screening from March 2004 to July 2014 in Obstetrics & Gynecology Hospital of Fudan University,were delivered and followed up until neonatal period on chromosome examination,prenatal B ultrasound and outcome of neonates.Receiver operating characteristics curves were plotted and used to predict the optimal critical point of poor prognosis and the warning point of follow-up.Results Among the 109 cases,83 cases tended to have a natural recovery during pregnancy,71 of them had normal renal pelvis.Two of the neonates died,9 cases needed surgical treatment,and 98 cases had normal renal pelvis or need follow-up only.The area under the receiver operating characteristics curves was 0.860 (95%CI:0.860± 1.96 × 0.112).The optimal critical point of poor prognosis was determined at ≥ 11 mm in anteroposterior diameter with sensitivity 81.8%,and specificity 83.7%,and the warning point of prenatal follow-up was at ≥ 7 mm in anteroposterior diameter with sensitivity 100.0% and specificity 50.0%.Conclusions The prognosis of isolated fetal pyelectasis is mostly good.The fetus with pyelectasis thicker than 7 mm should be followed-up closely during prenatal and neonatal period,and the fetus with pyelectasis thicker than 11 mm is likely to have poor prognosis in neonatal period.
7.MeaSurement of Relative Affinity and Specificity of Monoclonal Antibodies
Chun-Hai HAO ; Yuan-Yao CHEN ;
Chinese Journal of Immunology 1985;0(05):-
To measure the relative affinity and type Specificity of neutralizing monoclonal antibodies against the hexon antigen of adenovirus type 7,a simple ELISA double antibody binding system has been developed.By this method,ralative affinity of nine MAbs was estimated from the antibody concentrations at approximately 50% of plateau binding and were ranked.The theoretical basis for this method was discussed. Hybridomas secreting antibodies of desired affinity can be selected at an early stage after fussion by measuring relative affinity of hybridomas supernatants.The type specificity was judged by the difference between end-point concentration against adeno- virus type 7 and type 3.And the relation between affinity and specificity of MAbs was discussed.
8.Effects of SPARC on expression of TGF-βl and collagen type Ⅰ in keloid fibroblasts
Chun WANG ; Xiaojing LI ; Chaohua LIU ; Xiaojuan WENG ; Dongyue HAO
Chinese Journal of Medical Aesthetics and Cosmetology 2011;17(5):377-380
Objective To observe the effects of secreted protein,acidic and rich in cysteine (SPARC) on the expression of TGF-β1 and collagen type Ⅰ in cultured human keloid fibroblasts by real-time fluorescence quantitative RT-PCR.Methods In vitro keloid fibroblasts were stimulated by different concentrations of recombinant human SPARC,and with the control group for comparison,real-time fluorescence quantitative RT-PCR to detect expression of TGFβ1 and collagen type Ⅰ.Results Compared with the control group,the expression of TGF-β1 and collagen type Ⅰ was significantly increased in the experimental group.Conclusions SPARC could enhance the expression of TGF-β1 and collagen type Ⅰ in keloid fibroblasts significantly.
9.Relationship between characteristics of drug using in female drug users and double infection of hepatitis B virus and hepatitis C virus
Chun LIU ; Minzhu HUANG ; Lan GUAN ; Hao LIU
Chinese Journal of Tissue Engineering Research 2005;9(8):250-251
BACKGROUND: Drug use is the most significant risk factor for viral hepatitis B and C infection. Female drug users present different drug-using characteristics from the other sex. And what about the influence of these characteristics on hepatitis B and C virus(HBV and HCV) double infection?OBJECTIVE:To explore the relationship of characteristics of drug using in female drug users on the incidence of HBV and HCV double infection.DESIGN: It is a cross section investigation of female drug users in January 2002.SETTING: A laboratory separtment, an epidemiological department in a university and a drug dependency treatment unit.PARTICIPANTS: A total of 320 female drug users who were in Baimalongdrug dependency treatment unit in Zhuzhou city were recruited in this study.METHODS: The drug-using characteristics were investigated through a questionnaire and the surface antigen of HBV(HBsAg) and antibody against HCV(anti-HCV)in serum were checked with ELISA.MAIN OUTCOME MEASURES: Relationship between HBV and HCV infection and injection of drug, injecting career and abstaining history.RESULTS:The incidence of HBV and HCV double infection in the 320 female drug users was 9. 37%. All of those with double infection were injection drug users, which was different from non-injection drug users(x2 = 6. 276,P < 0. 05 ) . Longer career of drug-using meant higher possibility of double infection. The incidence of double infection (10. 3% ) in those of 5 - 10 years of drug-using career was higher than that (4.5%) of 1 - 5 years of drug-using career (x2 = 8. 344, P < 0.05) . And that in those experienced more than 4times of abstaining was also higher than that less than in those experienced 3times (x2 =9. 418, P < 0. 05) . Long career of drug using and repeated abstention abstaining were factors that leaded to HBV and HCV double infection (OR = 1. 923, 1. 597, 95% CI = 0. 964 - 3. 838, 0. 962 -2. 652).CONCLUSION: Injection drug use is the predominant factor that causes double infection of HBV and HCV and long career of drug using and rising in the number of times of abstaining also increase the double infection.
10.Relationship between sexual behavioral characteristics and co infection of hepatitis B virus and hepatitis C virus in drug abusers
Chun LIU ; Minzhu HUANG ; Lan GUAN ; Hao LIU ; Weiping DAI
Chinese Journal of Tissue Engineering Research 2005;9(4):236-237
BACKGRUND:The high prevalence of hepatitis B virus(HBV) is related with the sexual characteristics in drug abusers.How about the relationship between prevalence of HBV/hepatitis C virus(HCV) co infection and the sexual characteristics in drug abusers? OBJECTIVE:To investigate the relationship between sexual characteristics and HBV/HCV co infection in drug abusers. DESIGN:A cross sectional survey taking the drug abuser as the subjects. SETTING:A department of clinical laboratory of a municipal hospital,an epidemiological department of the college of public health of a university,a professional drug abuser treatment center. PARTICIPANTS:Totally 452 male and 320 female drug abusers were randomly selected from the Changsha Xinkaipu Drug Abusers Treatment Center and Zhuzhou Baimalong Female Drug Abusers Treatment Center as the subjects from 1st to 31st January 2001. INTERVENTIONS:A self designed questionnaire was used to investigate the sexual characteristics of drug abusers.HBV surface antigen and HCV antibody were detected in the serums of the participants with ELISA. MAIN OUTCOME MEASURES:The relationship between HBV/HCV co infection and the sexual behavior, numbers of sexual partners,and the usage of condom in drug abusers. RESULTS:The total prevalence of HBV/HCV co infection in drug abusers was 21.24% ; the prevalence of HBV/HCV co infection in male drug abusers was 29.6% ),which was significantly higher than that in females(9.4% )(χ 2=46.016,P< 0.01).There was no significant difference in the prevalence of HBV/HCV co infection between male and female drug abuser whether they had or had not sexual behaviors(χ 2=3.775,P >0.05).The prevalence of HBV/HCV co infection was in an ascending tendency with the increase of sexual partner(χ 2=9.846,P< 0.05).The using frequency of condom was significantly higher in females than in males(U=14.80,P< 0.01). CONCLUSION:There is no correlation between sexual experience and prevalence of HBV/HCV co infection;male drug abusers have higher prevalence of co infection than females;the number of sexual partners is correlated with the prevalence of HBV/HCV co infection,the more sexual partners,the higher prevalence is.The protective effect of condom is greater in female drug abusers than in males.