To investigate the induction of apoptosis of mouse colonic adenocarcinoma CT26 cells by recombinant Clostridium difficile toxin B (rTcdB), CT26 cells were exposed to different concentrations of rTcd B. Inhibition of cell proliferation was measured by MTT assay. The activation of Caspase 3 was measured by colorimetric method. Cell morphological analysis and flow cytometry were performed to confirm cell apoptosis. rTcd B inhibited the proliferation of CT26 cells in a timeand dose-dependent manner. Caspase 3 activity in CT26 cells was elevated remarkably after rTcd B exposure for 6 h, 12 h, 18 h or 24 h, as compared with the control group. Morphological changes were observed by fluorescence microscopy. The exposure of rTcd B to CT26 cells induced a timeand dose-dependent apoptotic cell death as determined by flow cytometry analysis. The results showed that recombinant Clostridium difficile toxin B induced apoptosis of CT26 cells.

0.05).P300 and amplitudes of the mismatch negativity(MMN) of group A were lower than that of group B.PL of MMN was obviously prolonged.By comparing PL of group A with thar of group B,there was obvious difference(P

Objective To construct a eukaryofic expression vector for A PP695 gene carrying green fluorescence protein (GFP) gene. Methods RT-PCR was used to amplify the full-length APP695 cDNA. The PCR products and the eukaryotic expression vector pIRES2-EGFP were digested by restriction endonueleases, and the digested APP695 gene was inserted into the digested eukaryotic expression vector. The positive recombinants were identified by PCR analysis, Nhe Ⅰ and Hind Ⅲ digestion and sequence analysis. Results The 2088-bp DNA fragment was amplified by PCR from the plasmid pCB6, and an identical DNA fragment was also amplified from the recombinants. The products of double restriction enzyme digestion were A PP695 gene with a 5.3-kb DNA fragment. Sequence analysis confirmed successful insertion of A PP695 gene into pIRES2-EGFP vector. Conclusion The eukaryotie expression vector pIRES2/APP695-EGFP has been successfully constructed.

Aspirin ; therapeutic use ; Diagnosis, Differential ; Female ; Humans ; Immunoglobulins, Intravenous ; therapeutic use ; Infant ; Male ; Mucocutaneous Lymph Node Syndrome ; diagnosis

Objective To explore the effect of dietary categories on effectiveness of the intestinal cleanliness before the colonoscopy,patients' acceptance of dietary changes and adverse reactions in bowel preparation.Methods Totals of 94 patients who underwent colonoscopy were divided into normal diet group (n =30),semi-liquid food group (n =32) and fasting group (n =32) according to the dinner diet styles the day before the colonoscopy.Observed three groups of adverse reactions,patients' acceptance and intestinal cleanliness.Results No significant difference was found in the medicine dosage,water intake,intestinal cleanliness among the three groups (P ＞ 0.05).The cases of fatigue phenomenon and discomfort on dietary changes in normal diet group were 4 and 1,in semi-liquid food group were 14 and 8,in fasting group were 13 and 5,and the difference was statistically significant (x2 =7.765,20.623,respectively; P ＜ 0.05).While no significant difference was found in the incidence of other adverse reaction among the three groups (P ＞ 0.05).Conclusions Dietary categories are no effect on the intestinal cleanliness of the colonoscopy.However,according to the conditions of the patients before taking laxatives poly would increase their comfort under certain dietary restrictions.

Objective To establish and test the reliability and validity of the disease related psychological pressure scale of pregnant women with chronic HBV infection,so as to provide mental stress assessment tool for the pregnant women.Methods Item pool was developed after the interview and comprehensively retrieved literatures,then after expert review and rebuilt,the scale items were fixed.Totals of 368 pregnant women with chronic HBV infection disease were investigated and the reliability and validity of scale were test.Results The scale had 5 factors and 24 items,and the correlation coefficient between each factor score and total score was 0.712 - 0.894.The correlation coefficient among each factor was 0.409 - 0.631 which was significantly lower than that between each factor score and total score ( P ＜ 0.01 ).The cumulative contribution rate of 5 factors was 64.055％,and tes-retest reliability of 5 factors was 0.856,0.887,0.828,0.813.The Cronbach' s α of internal consistency of 5 factors was 0.788 - 0.865,and the Cronbach' s α of scale was 0.932.Conclusions The scale has good reliability and validity,and can be used for measuring the disease related pregnancy psychological pressure for chronic HBV infection pregnant women.

Aim To investigate the influence of inhibi-tory nanocomposite on EC-9706 cells and the effect of nanocomposite on ESCCAL _ 1 LncRNA expression, siRNA-loaded nanocomposite being prepared as non-vi-rus delivery system Methods Mesoporous silica nano-particles were prepared by sol-gel method under room temperature and coated by cationic polymerpolyethylen-imine (PEI)on the surface to stay positive charge, which could facilitate its combination with negatively charged ESCCAL _ 1 siRNA. The size and surface charge of nanocomposite were determined by laser par-ticle analyzer and TEM. The inhibitory rate of nanopar-ticles on EC-9706 cells was detected by MTT methods. Entrapment efficiency was determined by agarose gel e-lectrophoresis. The uptake-siRNA was detected by flu-orescence microscope. The expression of ESCCAL _1 LncRNA was detected by RT-PCR. Results The MSNP appeared to have a high dispensability and hom-ogeneous size by particle size analyzer and transmission electron microscopy(TEM). The formed nanoparticles had a surface mesoporous diameter of 3 ~ 5 nm. The proliferation of ESCCAL_1 was inhibited significantlly (P < 0. 05),and the 72h inhibitory rate was (54. 93 ± 2. 6)%;the siRNA loading could be effectively up-taken by EC-9706 cells;ESCCAL_1 silencing efficien-cy was 69. 5% . Conclusions The tumor targeting nanocomposite with high encapsulation efficiency is prepared. The proliferation of esophageal cancer EC-9706 cells can be effectively inhibited by anocompos-ite-mediated siRNA,and the expression of ESCCAL_1 is effectively silenced in EC-9706 cells. The nanocom-posite is an efficient gene delivery system and may have potential application in gene therapy.

OBJECTIVETo explore a technology for diagnosing VHL mutations from a single cell and provide experimental evidences for the feasibility of applying technology in detecting genetic mutations from a single cell.

METHODSAfter whole genome amplification (WGA) based on multiple displacement amplication (MDA) for a single cell, we did regular PCR following sequencing and detected the genotypes using the real time PCR based on TaqMan probes. We detected VHL mutations by the different terminal fluorescent changing.

RESULTSThe rate of amplification for single cell based on MDA was 90.91%. The rate of contamination was 0. After sequencing, the allele drop out (ADO) rate of heterozygotes was 26.67%(8/30); combined with the different terminal fluorescent changing, the rate of ADO of heterozygotes was 16.67%.

CONCLUSIONWGA based on MDA for a single cell followed by regular PCR with sequencing and real time PCR can specially and accurately detect the VHL genotypes of single cells.

Base Sequence ; DNA Mutational Analysis ; Female ; Genotype ; Humans ; Lymphocytes ; metabolism ; pathology ; Middle Aged ; Molecular Sequence Data ; Mutation ; Polymerase Chain Reaction ; Preimplantation Diagnosis ; Von Hippel-Lindau Tumor Suppressor Protein ; genetics ; von Hippel-Lindau Disease ; blood ; genetics

OBJECTIVESTo study the effects of trichostatin A (TSA) on protein-protein interaction between HBx and histone deacetylase protein 1 (HDAC1).

METHODSBoth HBx and HDAC1 expressing vectors were constructed by the method of routine molecular cloning. The expression of HBx and HDAC1 were observed by Western blot assay. The protein-protein interaction was tested between HBx and HDAC1 by GST pull-down in vitro as well as co-immunoprecipitation in vivo.

RESULTSBoth HBx and HDAC1 expressing vectors were successfully constructed. Protein-protein interaction between HBx and HDAC1 existed both in vitro and in vivo. TSA, an inhibitor of HDAC1, had no effect on the interaction between HBx and HDAC1.

CONCLUSIONSHBx interacts with HDAC1 in vivo and in vitro in a non- TSA dependent way.

Histone Deacetylase 1 ; metabolism ; Humans ; Hydroxamic Acids ; metabolism ; Immunoprecipitation ; Plasmids ; Protein Interaction Mapping ; Trans-Activators ; metabolism

Objective To assist the office workers with pre-diabetes to get rid of the unhealthy lifestyle by living behaviour intervention and to analyze the effect on them.Methods Totally 50 patients were selected who meet the standards of diabetes in the early stage after having medical check up.Following measurements were carried out:formulating healthy diets and exercising plans purposefully,holding special lectures periodically and handing out brochures on health education.Results Unhealthy life behaviour was greatly improved after being intervened for one year.The differences of fasting blood glucose and the blood glucose two hours after a meal were statistically difference between pre-and post-intervention.Conclusions The living behaviour intervention can obviously lower the danger of pre-diabetes and improve the cases' life quality.