Construction of the full length and N-terminal fragment of serum response factor over-expressing lentiviral plasmid and its impacts on cardiac stem cell differentiation / 临床与实验病理学杂志

Purpose To analyze the effects of full length and N-terminal fragment of serum response factor (SRF-Full and SRF-N) on TGF-β1-induced differentiation in c-Kit + cardiac stem cells (CSC).Methods Rat SRF-Full and SRF-N (1-254 aa) coding sequences were obtained from cDNA library and cloned into the linearized lentviral vector GV358 (Ubi-MCS3FLAG-SV40-EGFP-IRES-puromycin) to generate the recombinant vectors,and then positive clones were selected and sequenced after transducing the competent cells with recombinant vectors.The recombinant lentvirus were packaged through transfecting the HEK293T cells with SRF-Full,SRF-N overexpressing plasmids and viral packaging plasmids.Neonatal SD rat cKit + CSCs were isolated via magnetic activated cell sorting,and TGF-β1-induced differentiation in SRF-Full and SRF-N overexpression virus-infected CSCs was assessed by quantitative PCR.Results SRF-Full and SRF-N coding sequences were successfully obtained and properly cloned into the linearized GV358.The positive clones were selected and further confirmed by sequencing.With the help of packaging plasmids,the SRFFull and SRF-N overexpressing plasmids-transfected HEK293T cells successfully produced the lentiviral particles with the titer of 2 × 108 TU/mL,and the SRF-Full-Flag and SRF-N-Flag fusion protein were detected by Western blot in virus-infected HEK293T cells.Addition of TGF-β1 significantly induced upregulated mRNAs in cardiomyocyte markers (Nkx2.5,Gata4,cTnI) and smooth muscle cell marker (SM22α) but not the epithelia cell marker (vWF) in CSCs.Overexpression of SRF-Full facilitated TGF-β1-triggered cardiomyocyte differentiation.However,SRF-N exerted anti-differentiation effects in TGF-β1-treated cells.Conclusion The SRF-Full and SRF-N overexpressing recombinant lentiviral vectors are successfully constructed.SRF-Full facilitates while SRF-N suppresses TGF-β1-induced cardiomyocyte differentiation in c-Kit + CSCs.

A study on the reliability and validity of the Chinese revised version of Barratt Impulsiveness Scale in Chinese college student with a web-based survey / 预防医学

Objective TheaimofthestudywastoexaminethepsychometricpropertiesoftheChineserevisedversionof BarrattImpulsivenessScale11th(BIS-11)inChinesecollegestudentwithaweb-basedsurvey.Methods Atotalof 2 295 college students were enrolled in the survey,and were divided into two groups.The first group was used for item and factoranalysis,andthesecondgroupwasusedforconfirmatoryfactorsanalysis.Results ItemanalysisindicatedthatBIS-11 had satisfactory item discrimination,except the item 29.Three -factor model of BIS -11 was well documented with exploratory factor analysis (explained 45.526%of total variance)and confirmatory factor analysis (GFI,AGFI,TLI,CFI, RMSEA was 0.872,0.851,0.853,0.864,0.064,respectively).The internal consistency of the total scale and the three subscales using coefficient alpha was in the range of 0.833-0.913.The split-half reliability of the total scale and the three subscales using Spearman-Brown Coefficient was in the range of 0.827-0.907.Furthermore,the female college students in the present study had higher scores on the total scale,cognitive (attention)impulsiveness factor,and motor impulsiveness factor than the male college student (P <0.01 ).The individuals with GHQ -12 (the twelve -item General Health Questionnaire)screen-positive had higher scores on the total scale and the three factors than the subjects with GHQ-12 screen-negative(P<0.01).Conclusion TheresultsofpresentstudysuggestedthattheChineserevisedversionofthe Barratt Impulsiveness Scale 1 1 th could be used as a tool of impulsiveness assessment in web-based survey.

Inhibition of Pim-1 attenuates the proliferation and migration in nasopharyngeal carcinoma cells

OBJECTIVE@#To explore the role of proto-oncogene Pim-1 in the proliferation and migration of nasopharyngeal carcinoma (NPC) cells.@*METHODS@#Pim-1 expressions in NPC cell lines CNE1, CNE1-GL, CNE-2Z and C666-1 were examined by RT-PCR, western blotting and immunoflucesence, respectively. After CNE1, CNE1-GL and C666-1 cells were treated with different concentrations of Pim-1 special inhibitor, quercetagetin, the cell viability, colony formation rate and migration ability were analyzed.@*RESULTS@#Pim-1 expression was negative in well-differentiated CNE1 cells, whereas expressed weakly positive in poor-differentiated CNE-2Z cells and strongly positive in undifferentiated C666-1 cells. Interestingly, CNE1-GL cells that derived from CNE1 transfected with an Epstein Barr virus latent membrane protein-1 over-expression plasmid displayed stronger expression of Pim-1. Treatment of CNE1-GL and C666-1 cells with quercetagetin significantly decreased the cell viability, colony formation rate and migration ability but not the CNE1 cells.@*CONCLUSIONS@#These findings suggest that Pim-1 overexpression contributes to NPC proliferation and migration, and targeting Pim-1 may be a potential treatment for anti-Pim-1-expressed NPCs.

Study on the Optimum Culture for Growth of Bacillus Subtilis B115 andthe Antibacterial Effect of B115 on Aeromonas / 微生物学通报

The essential medium of B115 composed of 1% tryptone, 0.25% yeast extract and 0.5% sodium chloride was determined by using an orthogonal design. The orthogonal design was also employed in testing the optimum additions. It was composed of 0.1%(NH_(4))_(2)SO_(4),1.4%K_(2)HPO_(4), 0.6% KH_(2)PO_(4) and 0.1% (Na_(3)C_(6)H_(5)O_(7)). The yield of B115 cultured in optimum medium was compared with the one in essential medium. Statistic analysis showed that the growth of B115 was most significantly improved by adding K~(+)、NH~+_(4) and (Na_(3)C_(6)H_(5)O_(7)) to essential medium. The antibacterial effect of Bacillus subtilis strain B115 on pathogenic Aeromonas was studied. The results showed different antibacterial effects of B115 on different aeromonads. There were obvious antibacterial effects on BSK-10 and CL990920, while no effect on the growth of TL970424.