Objective To upregulate Notch signaling in cancer cells by overexpression of active part of Notch1 and to examine the proliferation of the cells. Methods Four cancer cell lines were infected with retrovirus recombined with sequence encoding active part of Notch1.CBF-1 reporter plasmid was used to detect Notch signaling and proliferation assay was carried out by MTS method.Cell cycle analysis was synchronously conducted. Results The overexpression of the active part of Notch1 induced upregulation of Notch signaling,led to growth inhibition in Hela and HepG2 cell lines and growth boost in BGC-823 cell lines,while had no effect on Chang cell lines. Conclusion The upregulation of Notch signaling can exert various effects on different cancer cell lines which is critical to the gene therapy for cancers.

Objective To study the activity of the survivin gene promoter in several tumor cell lines and evaluate the possible application of this promoter in tumor gene therapy. Methods ①The expressions of survivin gene in A549,MDA-MB231 and HepG2 cell lines were detected by RT-PCR and Western blotting.②Tumor cells(A549,MDA-MB231,HepG2) were transiently transfected by reporter plasmids containing different length of survivin promoter using lipofectamine.And 48 h later,the level of reporter gene expression was analyzed.Results There were different levels of survivin expression in A549,MDA-MB231 and HepG2 cell lines.Transient transfection assay approved that pLuc-surP-987,pLuc-surP-596,pLuc-surP-269 and pLuc-surP-158 showed high activity and 269 bp survivin promoter demonstrated the highest activity. Conclusion In transcriptional level,survivin promoter can activate the reporter gene in several tumor cell lines.It is a potential candidate promoter in tumor gene therapy.

There are many different opinions about the taxonomy of plant pathogenic coryneform bacteria since they were departed from genus of Corynebacteria. In recent years, they were classified into 5 genus, including Clav-ibacter, Curtobacterium, Arthrobacter, Rhodococcus and Rathayibacter. Some new points of view about their taxonomy have been published thereafter. The changing of taxonomy is maily because of the methods'altering from old to new molecular and polyphasic taxonomy, and the latter is in continuously development. Taxonomy of plant pathogenic coryneform bacteria somehow depends on the cooperation of phytopathologists, microbilogists and other scientists.

Effect of organic acids on the synthesis of 1,3 propanediol was studied.The adsorption of organic acids from glycerol fermentation liquor by ion-exchange resins was investigated.The results showed that organic acid and 1,3 propanediol production was in negative relationship.The static adsorption showed that ion-exchange resin 005 had the best adsorption abilities of the organic acids in the glycerol fermentation liquor.It was showed that the yield of 1,3propanediol increased by 166% after the extraction of organic acids from glycerol fermentation liquor and the convertion rate increased by 34%.

Objective To investigate the influence of murine cytomegalovirus(MCMV) infection on differentiation and differentiation gene expression of neural stem cells (NSCs) in vitro for studying the mechanisms of brain abnormalities calmed by congenital cytomegalovirns infection. Methods NSCs were separated from fetal BALB/c mouse and cultured and identified in vitro. The differentiation potency of NSCs was observed by immunnfluorescence. The NSCs infected by MCMV at dosage of multiplicity of infection (MOI) equaled to 5, I and 0. 1, respectively, were cultured in differentiation medium. The morphological changes of the cells were observed by inverted microscope. The ratios of NSCs and its differentiated cells were detected by flow cytometry. The expression changes of nestin, GFAP and NSE, markers of NSCs and its differentiated cells, were studied by immunofluorescence ( MOI = 1 ). The expression of early antigen (EA) of MCMV was detected to observe the infection process. Real-time RT-PCR method was employed to measure the expression levels of the key differentiation genes Wnt-3 and Wnt-7a in Wnt signal pathway of NSCs at early phage of differentiation culture. Results NSCs isolated from embryonic mouse brains could proliferate to form neurnspheres and strongly express Nestin and differentiate into NF-200 positive neurons or GFAP positive astrocytes. The NSCs of the infected groups couldn't adhere to the wall and appear differentia-tion growth, but showed swollen gradually after differentiation culture. The nostin expression of the infected groups downregulated slowly and was higher than that of the control groups ( P < 0.05 ). The GFAP and NSE expression of the infected groups were lower than that of the control groups (P <0.05). The EA of MCMV could be always detected in the cells of the infected groups. The ratios of nestin positive cells of the infected groups were higher than that of the control groups, but the ratios of GFAP and NSE positive cells of the for-mer were lower than that of the latter from 3rd to 9th day after differentiation culture ( P < 0.05 ). The levels of Wnt-3 mRNA and Wnt-7a mRNA of the infected groups were markedly lower than that of the control groups from 1st to 2nd clay and from 12th hour to 2nd day after differentiation culture respectively ( P < 0.05 ) . These changes of the infected groups became more obvious as MCMV MOI increased . Conclusion MCMV could inhibit significantly NSCs differentiate to neurons and astrocytes and lead to the decrease of dif-ferentiated cells. MCMV could inhibit or interfere with the gene expression of Wnt-3 and Wnt-7a in Wnt sig-nal pathway of NSCs. The effect that MCMV inhibited the differentiation and the differentiation gene expres-sion of NSCs showed dose-dependent with MCMV MOI. The inhibitory effect of MCMV on the differentiation of NSCs might be induced by interfering the differentiation gene expression of NSCs, which is possibly the one of primary causes of brain development disorders caused by congenital CMV infection.

The purpose of this study was to explore the regulatory effect of thymosin α1 (Tα1) on expression of TOLL-like receptor 9 (TLR9)/indoleamine2, 3-dioxygenase (ido) mRNA in bone marrow mesenchymal stem cells (MSC) from children with aplastic anemia (AA). Culture system of bone marrow MSC from AA children and normal children in vitro was established, and the effects of Tα1 on expressions of tlr9 mRNA and ido mRNA of MSC from AA children and normal children were determined by RT-PCR. The results showed that the bone marrow MSC from normal children did not express tlr9 and ido mRNA. Bone marrow MSC from children with AA obviously expressed tlr9 mRNA , but did not express ido mRNA; AA children's MSC treated with Tα1 for 18 hours markedly down-regulated tlr9 mRNA expression, but up-regulated ido mRNA expression in the concentration- and time-dependent ways. It is concluded that Tα1 can up-regulate the expression of ido mRNA in bone marrow MSC from children with AA.
Adolescent ; Anemia, Aplastic ; metabolism ; Bone Marrow Cells ; drug effects ; metabolism ; Cells, Cultured ; Child ; Female ; Gene Expression Regulation ; HL-60 Cells ; Humans ; Indoleamine-Pyrrole 2,3,-Dioxygenase ; metabolism ; Male ; Mesenchymal Stromal Cells ; drug effects ; metabolism ; RNA, Messenger ; genetics ; Thymosin ; analogs & derivatives ; pharmacology ; Toll-Like Receptor 9 ; metabolism

Objective To investigate the clinical value of 99Tcm-MIBI gated G-MPI in detecting the myocardial damage in sarcoglycanopathy.Methods 99 Tcm - MIBI G - MPI was performed in 8 patients (3 males,5 females,age ranged from 10 to 30 y) with sarcoglycanopathy confirmed by clinical results and molecular pathology and 4 healthy persons as control group.Quantitative gated SPECT (QGS) software was used for processing and interpretation.Myocardium of left ventricle was divided into 7 segments and 20 subsegments.A five-point scoring system was used to evaluate the myocardial damage.Results Seven patients showed positive results in G-MPI (7/8).Fifty-nine sub-segments of injured myocardium were detected in the 140 sub-segments of 7 abnormal patients.One abnormal segment was observed in 1 patient,two abnormal segments were detected in 2 patients,and ≥3 abnormal segments were observed in 4 patients.Enlarged left ventricles were detected in 5 patients (5/8),and the LVEF of 3 patients among them decreased to (43.1 ±2.8)％.Conclusion 99Tcm-MIBI G-MPI can detect myocardial damage in sarcoglycanopathy as a direct and non-invasive method,and can be used in the early diagnosis and long-term follow-up in sarcoglycanopathy.

Drug reservation wag investigated in 2077 community residents.We found that most drugs were obtained from the hospitals(83.78%),kept at a relatively lower place(69.23%),packed in box(75.25%),and did not meet the storage requirement(72.60%).Half of the overdue drugs(median time,12 months)were used for internal treatment.This study suggests that there might be unsafe drug storage in communities.

mutants which didnt produce red pigmen ts on malt extract agar plate were obtained.The 8 stable mutants were cultured on solid medium.Two samples wer e yellow,the others were white.The extracted samples were scanned in visible len gth.2 yellow samples showed only one absorptive peak at 370nm,the 6 white sample s showed no absorptive peak.The mutants producing only yellow pigments on solid medium were tested in liquid culture.The results indicated their ability to pro duce only yellow pigments were stable.

OBJECTIVETo study the regulatory effect of thymosin α1 (Tα1) on immunosuppression of bone marrow mesenchymal stem cells (MSCs) from children with aplastic anemia (AA) through Toll-like receptor 9(TLR9)and indoleamine 2,3-dioxygenase (IDO) signaling pathway.

METHODBone marrow T cell subsets from children with AA and normal individuals were measured by using flow cytometry. Expressions of TLR9/IDO mRNA of MSCs cocultured with Tα1 were determined by reverse-transcription PCR (RT-PCR). Inhibition of PHA-activated T cell proliferation and activation by MSCs cocultured with Tα1 was detected by using MTT assay and flow cytometry.

RESULTCD4(+)/CD8(+) ratio (0.64 ± 0.02) in children with AA was significantly lower than that in normal individuals (1.42 ± 0.05); but CD8(+)/CD38(+) ratio (0.92 ± 0.04) was significantly higher than that in normal individuals (0.65 ± 0.05). AA MSCs obviously expressed TLR9, but not IDO; AA MSCs treated with Tα1 downregulated TLR9 expression but upregulated IDO expression in concentration- and time-dependent manners. The inhibition of AA MSCs on T cell proliferation (21.38% ± 12.34%) was lower than that in normal individuals (62.72% ± 17.79%, P < 0.05), while AA MSCs treated with Tα1 for 18 h exhibited a stronger inhibition (42.83% ± 16.54%, P < 0.05).

CONCLUSIONThe immunosuppression mediated by MSCs could be improved by Tα1 through upregulation of IDO expression via TLR9-dependent signaling pathway. This research provides a new idea for targeted immunomodulatory therapy with bone marrow MSCs from children with AA.

Adolescent ; Anemia, Aplastic ; metabolism ; Bone Marrow Cells ; drug effects ; metabolism ; Case-Control Studies ; Child ; Female ; Humans ; Immune Tolerance ; Immunosuppression ; Indoleamine-Pyrrole 2,3,-Dioxygenase ; metabolism ; Male ; Mesenchymal Stromal Cells ; drug effects ; metabolism ; Signal Transduction ; immunology ; Thymosin ; analogs & derivatives ; pharmacology ; Toll-Like Receptor 9 ; metabolism