Objective To investigate mutations in immediate early ( IE) gene ORF62 of three var-icella vaccine Oka strains ( vOka ) including two strains produced in China and their parental Oka strain (pOka), and then to further elucidate its possible roles in attenuation mechanism by comparing their ORF 62 promoter sequences and its activities , ORF62 coding regions and its transactivities .Methods ORF62 pro-moter-reporter plasmids and ORF62-expressing plasmids of pOka and three vOka strains ( vOka-BK from Changchun BCHT Biotechnology Co ., vOka-SH from Shanghai Institute of Biological Product Co .Ltd., and vOka-GSK from GlaxoSmithKline plc, as control) were constructed, respectively.ORF62 promoter regions and coding regions of the four strains were sequenced and then compared with each other .Differences of ac-tivities of the ORF62 promoter, and transactivities of the ORF62-encoded IE62 upon immediate early (ORF4), early (ORF28) and late (ORF67) gene promoters between pOka and vOka strains were assayed with transient transfection technique .Results Compared with pOka strain , three vOka strains had a con-sistent T deletion mutation at site 110 050 in ORF62 promoters, which did not result in any change of tran-scription factor binding motif .However , activities of ORF62 promoters from three vOka strains were signifi-cantly lower than those of pOka strain .Three consistent substitution mutations were observed in ORF 62 cod-ing regions of three vOka strains and three new enzyme restriction sites including SmaⅠ, NaeⅠand BssHⅡwere generated, respectively.Transactivities of IE62 from three vOka strains upon ORF4, ORF28 and ORF67 promoters were significantly higher than those of pOka both in CV-1 and MeWo cells , except that vOka-SH IE62 showed significantly lower transactivities upon ORF 4 promoter than those of pOka strain in CV-1 cells.Conclusion Consistent T deletion mutation at site 110 050 in ORF62 promoters of three vOka strains might be responsible for the reduced promoter activities and the changes of IE 62 transactivities .How-ever , it seemed that cell types have no significant effect on ORF 62 promoter activity or IE 62 transactivity be-tween pOka and vOka strains .

In order to study the effects of c-myc antisense phosphorothioate oligodeoxynucleotide in inducing apoptosis of HL-60 cells, the expression of c-myc mRNA was determined by RT-PCR, the morphologic signs of apoptotic cells were observed by transmission electron microscopy, the proportion of apoptotic cells was detected by flow cytometry, and the DNA fragments were analysed by agarose gel electrophoresis. Results of RT-PCR showed marked decrease of c-myc mRNA expression in AspoI and II treated cells. The level of c-Myc protein was decreased 23.8% and 45.4%, respectively, in cells treated with 5 and 10 micro mol/L AspoI, and 38.4% after treatment of 10 micro mol/L AspoII. The apoptotic rates were 23.97% and 52.6% after 10 micro mol/L AspoI treatment and 28.8% and 45.19% after treatment with AspoII 10 micro mol/L for 48 and 72 hours, respectively. while apoptotic cells did not apear in control and sense oligodeoxynucleotide groups. Electron microscopy observation showed the characteritics of apoptosis. A ladder like pattern of DNA fragments was demonstrated on electrophoretogram. The results suggst that c-myc antisense oligodeoxynucleotides could be highly specific gene agents that can suppress the level of c-myc mRNA, decrease the c-Myc proteins and induce apoptosis of HL-60 cells.

BACKGROUNDInterleukin 10 (IL-10) is an important cytokine with anti-inflammatory, anti-immune and anti-fibrotic functions. This study aimed at evaluating the relationship between allele polymorphisms in the IL-10 promoter region and hepatitis B virus (HBV) or hepatitis C virus (HCV) infection.

METHODSThe odds ratios (ORs) of IL-10 allele distributions in patients with HBV or HCV infection were analyzed against healthy controls. All the relevant studies in PubMed were identified, and poor qualified studies were excluded. The meta-analysis software REVMAN 4.2 was applied for investigating heterogeneity among individual studies and summarizing all the studies. The publication bias was also evaluated.

RESULTSThis study demonstrated a significant association between the IL-10-592 A/C polymorphism and HBV infection in the Asian population under the overall effect size of allele A versus C. In our subgroup meta-analysis, we found a significant association of IL-10-592 A/C polymorphism to HCV infection susceptibility in Asian populations, although sensitivity analysis showed that the combined result was not associated with the worldwide population. Other IL-10 allele polymorphisms were not associated with HBV or HCV infection.

CONCLUSIONIL-10-592 A/C allele might be a risk factor for HBV or HCV in Asians but not in Europeans.

Alleles ; Genetic Predisposition to Disease ; genetics ; Hepatitis B ; epidemiology ; genetics ; Hepatitis C ; epidemiology ; genetics ; Humans ; Interleukin-10 ; genetics ; Polymorphism, Genetic ; genetics ; Promoter Regions, Genetic ; genetics

Bilingual teaching is adapted to the development of higher education in china.Based on actual fact of college,teaching mode,evaluation and effect of bilingual teaching on medical microbiology were studied,which started with necessity of bilingual teaching to use original edition teaching material in English. The result would provide some gist to choice the suitable pattern of bilingual teaching for other subject of our college.

The fused tooth is the union of two dental enamel or dentin formed together. In the maxillary, the fusion usually occurred within the lateral incisor and canine and very rarely occurred in the upper third molar and supernumerary tooth. This paper reported a fused tooth occurred in the left maxillary impacted third molar with supernumerary tooth.
Fused Teeth ; Humans ; Incisor ; Male ; Maxilla ; Molar ; Molar, Third ; Tooth, Impacted ; Tooth, Supernumerary

The stem cell leukemia (SCL) gene is a new oncogene related with leukemogenesis. To explore the effects of antisense oligonucleotides of SCL on leukemic cells, SCL antisense phosphorothioate oligodeoxynucleotides (AS-PS-ODN) were used to treat K562 and CEM leukemic cell lines to observe the effects on proliferation, differentiation, apoptosis and SCL mRNA expression in the cells. The results showed that incubation of K562 or CEM cells with AS-PS-ODN at different concentrations led to inhibition of cell proliferation, and the inhibitory effects varied with the incubation time. The positive rate of benzidine staining in K562 cells increased significantly after the inhibition with AS-PS-ODN, compared with S-PS-ODN treatment. The characteristics of apoptosis were observed in K562 cells treated with AS-PS-ODN, but not in CEM cells. Expression of SCL mRNA in K562 and CEM cells and SIL-SCL mRNA in CEM cells decreased after incubation of AS-PS-ODN. It is concluded that SCL AS-PS-ODN inhibits specifically the proliferation of K562 and CEM cells, also decreases the level of SCL and SIL-SCL mRNA expression. AS-PS-ODN enhances erythroid differentiation and induces premature apoptosis in K562 cells.
Apoptosis ; drug effects ; Basic Helix-Loop-Helix Transcription Factors ; Cell Division ; drug effects ; DNA-Binding Proteins ; antagonists & inhibitors ; physiology ; Humans ; K562 Cells ; Oligonucleotides, Antisense ; pharmacology ; Proto-Oncogene Proteins ; antagonists & inhibitors ; physiology ; RNA, Messenger ; analysis ; T-Cell Acute Lymphocytic Leukemia Protein 1 ; Transcription Factors ; antagonists & inhibitors ; physiology

OBJECTIVETo study the effect of endogenous TGF-beta(1) and TNF-alpha on As(2)O(3) inducing apoptosis of HL-60 cells.

METHODSThe expressions of endogenous TGF-beta(1) and TNF-alpha in apoptotic HL-60 cells induced by As(2)O(3) were assayed by RT-PCR, quantitative RT-PCR, ELISA, DNA fragmentation and TUNEL. The effect of TGF-beta(1) and TNF-alpha antisense phosphorothioate oligodeoxynucleotides (PSODNs) on As(2)O(3) inducing apoptotic HL-60 cells was further studied.

RESULTS(1) Expressions of endogenous TGF-beta(1) and TNF-alpha were significantly up-regulated in As(2)O(3) inducing apoptotic HL-60 cells (from 13,546 +/- 124 and 497,216 +/- 187 before treatment to 23,273 +/- 229 and 674,217 +/- 189 after treatment, respectively), accompanied with down-regulated bcl-2 mRNA expression (from 10,424 +/- 274 before treatment to 3,361 +/- 89 after treatment). (2) TGF-beta(1) and TNF-alpha antisense PSODNs could rescue As(2)O(3) induced apoptosis of HL-60 cells, with a restoration of bcl-2 gene expression.

CONCLUSIONSEndogenous TGF-beta(1) and TNF-alpha played an important role in As(2)O(3) inducing HL-60 cells apoptosis through down-regulation of bcl-2 expression.

Antineoplastic Agents ; pharmacology ; Apoptosis ; physiology ; Arsenicals ; pharmacology ; Down-Regulation ; HL-60 Cells ; Humans ; Oxides ; pharmacology ; Proto-Oncogene Proteins c-bcl-2 ; biosynthesis ; genetics ; RNA, Messenger ; genetics ; Receptors, Tumor Necrosis Factor ; metabolism ; Transforming Growth Factor beta1 ; biosynthesis ; physiology ; Tumor Necrosis Factor-alpha ; physiology

Objective:To investigate risk factors for vascular cognitive impairment(VCI)in elderly patients 12-18 months after the onset of acute partial anterior circulation infarction(PACI), and to establish a diagnostic and predictive model.Methods:This was a prospective study. Demographic characteristics, vascular risk factors and laboratory data of 148 patients with acute PACI were collected, and patients were followed up for 12-18 months.The Montreal Cognitive Assessment Scale(MoCA)was used to evaluate patients' cognitive function.Logistic stepwise regression was used to screen risk factors for VCI.We established a diagnostic and predictive model.The area under the receiver operating(ROC)curve(AUC)was used to evaluate the efficiency of the model.Results:A total of 126 subjects completed the 12-18 month follow-up.Multivariate logistic regression analysis found that high homocysteine(Hcy)( OR=1.082, 95% CI: 1.002-1.167), high glycated hemoglobin(HbA1c)( OR=1.653, 95% CI: 1.052-2.598), high National Institutes of Health Stroke Scale(NIHSS)score( OR=1.291, 95% CI: 1.098-1.518), high hypersensitive C-reactive protein(hs-CRP)( OR=1.026, 95% CI: 1.005-1.047)and low education level( OR=2.485, 95% CI: 1.231-5.018)were independent risk factors for VCI in patients 12-18 months after PACI( P<0.05). The AUC of the diagnostic and predictive model was 0.828(95% CI: 0.755-0.902). Conclusions:High Hcy, NIHSS score, hs-CRP and low education level are independent risk factors for VCI in patients 12-18 months after PACI.The diagnostic and predictive model can help to screen patients at high-risk for VCI, so that timely clinical recognition, diagnosis and treatment can be made after acute PACI.

To investigate the effects of wt-p53 gene on proliferation and differentiation of K562 cells and to explore the feasibility of wt-p53 in leukemia gene therapy, pC53-SN(3), containing wt-p53 cDNA, and temperature-sensitive p53 mutant pN53cG(Val135) which behaved like wt-p53 at 32.5 degrees C, were introduced into p53-null K562 cells respectively by lipofectin mediated DNA transfection. In the presence of G418, K-SN(3) and K-pN53cG clones expressing P53 protein were selected. The effects of exogenous wt-p53 gene on the proliferation and differentiation of K562 cells were studied by detection of cell growth curves, leukemic colony formation, cell cycle analysis and DNA fragmentation, TdT-mediated dUTP nick end labeling (TUNEL) and benzidine staining. The results showed: (1) The level of p53 mRNA in K-SN(3) cells was lower than that in K-pN53cG cells by RT-PCR. (2) K-SN(3) and K-pN53cG(32.5 degrees C) cells proliferated more slowly than the control K562 cells, and their colony formation was obviously suppressed. The cells in G(0)/G(1) phase increased, and the cells in S phase decreased. These features were more obvious in K-pN53cG(32.5 degrees C). (3) K-pN53cG(32.5 degrees C) showed the feature of apoptosis and K-SN(3) showed the characteristics of erythroid lineage differentiation. It was indicated that exogenous of wt-p53 was capable of inhibiting the proliferation of K562 cells and inducing apoptosis of the cells at higher p53 level and interestingly, inducing the cells differentiation on erythroid lineage at lower p53 level.

To investigate whether F951, a novel bcl-2 antisense oligodeoxynucleotide, increases the sensitivity of HL-60 cells to Ara-C, HL-60 cells were cultured with F951 in different doses alone or with F951 combined with low-dose Ara-C; the proliferation of HL-60 cells was assayed by MTT and trypan blue exclusion test; expression of Bcl-2 protein and its mRNA were measured by FACS and RT-PCR, respectively; the apoptotic cells were detected by DNA ladder and TUNEL assay. The results showed that F951 combined with low dose Ara-C revealed stronger effects in the aspects of inhibiting the HL-60 cells proliferation than in different doses of F951 alone or Ara-C alone. HL-60 cells treated with F951 + Ara-C had significantly lower trypan blue exclusion rate than that treated with Ara-C alone. The inhibition rates of HL-60 cells treated with FNS, Ara-C, F951 and F951 + Ara-C were -2.8%, 27.63%, 37.66%, 57.24%, respectively. F951 significantly down-regulated the expression of bcl-2 mRNA and protein in HL-60 cells. HL-60 cells treated with F951 + Ara-C showed more apparent DNA ladder and more apoptotic cells. It is concluded that F951 can inhibit bcl-2 gene expression and enhance the cytotoxicity of Ara-C through promoting apoptosis in HL-60 cells, hence increases the antitumor effect of Ara-C.
Antimetabolites, Antineoplastic ; pharmacology ; Apoptosis ; drug effects ; Cell Proliferation ; drug effects ; Cytarabine ; pharmacology ; Dose-Response Relationship, Drug ; Drug Synergism ; Flow Cytometry ; HL-60 Cells ; Humans ; Oligodeoxyribonucleotides, Antisense ; genetics ; pharmacology ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction