Objective: To establish an in vitro screening system for testing effects of MAD2 gene of embryonic cell genome.Methods: In order to construct the plasmid expressing MAD2-GFP fusion plasmid,the target gene fragment was obtained by RT-PCR amplification and inserted into pEGFP-N1 reporter vector.The structure of constructed plasmid was confirmed by electrophoresis analysis and DNA sequencing.The function of construct was confirmed by lipofectamine-mediated transient expression in HepG2 cells.Results: DNA sequencing showed that the inserted fragment of the constructed plasmid was the same as the template MAD2 genome.The HepG2 cells transfected with the constructed plasmid could express reporter gene of gfp.Conclusions: the plasmid expressing gfp gene controlled by MAD2 gene is successfully constructed and an in vitro testing system for evaluating founction of MAD2 gene in separation of cell is established.

objective:To screen and identify the interactive proteins of Mps1 and provide clues for the studies on Mps1 function on the chromosome segregation.Methods:The human embryonic cDNA library was screened with pDBLeu-Mps1 as bait plasmid by yeast two-hybrid system.And two Leu+ and LacZ+ positive yeast clones were obtained,which were contransformed to the MaV203 along with pDBLeu-Mps1 plasmid one to one by yeast simultaneous contransformation so as to identify the protein interactions again.The inserted fragments of identiyied positive clones were sequence and analyzed with BLAST in NCBI.Results: Only one positive clone identified one to one yeast simultaneous cotransformation,and the other was negative.The positive clone fragment was MAD1 protein by BLAST analysis in NCBI.Conclusion:To screen and initially identify the interactive protein of.Mps1 this research is a foundation for the study on the founctions of Mps1.

Objective:To discuss the mechanism of down-regulated expression of MAD2 gene in trophoblastic cells by microRNA.Methods:Endogenous MAD2 mRNA level was compared by using quantitative real-time PCR before and after transfected with microRNA plasmid;and the Mad2 protein level was compared by western blot.Results:Ratio of MAD2 mRNA/GAPDH mRNA of control group was 0.1780?0.0688,and after transfected with microRNA recombinant plasmid was 0.1778?0.0689 and 0.1778?0.0670respectively.Statistical study of samples showed they have no instinct different;otherwise the protein level in one of the experimental groups was significantly decreased after transfected with microRNA plamid.Conclusion:This study established microRNA can down-regulate gene expression in mammalian cells at translation level,it is a good foundation to study on the control of gene expression.

Objective:To study the expression of mitosis checkpoint gene hsMAD2 in colorectal cancer. Methods:The expression of hsMAD2 gene mRNA in colorectal cancer and normal colorectal tissue of 18 cases was quantitatively detected by RT-PCR. Results:The ratio of hsMAD2 mRNA/ GAPDH mRNA in colorectal cancer and normal colorectal tissue were 0.1235?0.0752 and 0.0312?0.0598,respectively. The results suggested that the expression of hsMAD2 mRNA in colorectal cancer were significant higher than in normal colorectal tissue(P

Objective:To investigate the expression of Trophinin gene in the mouse endometrium during the period of blastocyst implantation.Methods:By reverse transcription-polymerase chain reaction(RT-PCR),Trophinin gene in the mouse endometrium of 1～6 day normal pregnancy was detected.Results:The Trophinin mRNA was not detected in endometrium of nonpregnant and 1～2 day pregnant mouse.But in 3～5 day normal pregnancy it was strongly expressed.The Trophinin gene was scarcely detected in the endometrium of the 6 day pregnancy.Conclusion:Expression time of Trophinin mRNA in mouse endometrium coincides with blastocyst implantation.This suggests Trophinin participate in the adhesion of blastocyst and endometrium.

Objective:To discuss the depressed expression of BUB1 gene in trophoblastic cells by microRNA.Methods:Endogenous BUB1 mRNA level was compared by using quantitative real-time PCR before and after transfected with microRNA plasmid; and the Bub1 protein level was compared by western blot.Results:Ratio of BUB1 mRNA/GAPDH mRNA of control group was 0.1960?0.0688,and after transfected with microRNA recombinant plasmid was 0.1968?0.0689 and 0.1962?0.0670 respectively.Statistical study of samples showed they have no instinct different;otherwise the protein level was significantly decreased after transfected with microRNA plamid.Conclusion:This study established microRNA can control gene expression in mammalian cells at translation level,it is a good foundation to study on the control of gene expression.

The construction of key laboratory in colleges is the important warranty of the upgrading of school's comprehensive research competence.In this text,we have discussed how to strengthen the building of key laboratory and facilitate the overall development of research system in university,and considered the essentiality of construction and the reason of development.

OBJECTIVE: It's practically significant to study college students' behavior and psychology in using the Internet so as to guide them to take advantage of the Internet properly and to promote the proper use of the Internet. DATA SOURCES: The related articles indexed in the Medline Databank from January 2000 to January 2004 were searched by using computer with the index words "students' psychology, computer communication netware, elements analysis", and the language of the articles was limited to English. Meanwhile, the related articles indexed in the Full-length Chinese Journals Databank, Wanfang Databank and the Journal of Chinese Clinical Rehabilitation from January 2000 to January 2004 were searched through the computer with the index words "students' behavior, internet behavior, analysis of elements ", and the language of the articles was limited to Chinese. The subjects of the study were the Chinese college students aged 18 -24.STUDY SELECTION: The materials were studied first to choose the literature on the analysis of the collage students' Internet psychology and behavior. Inclusion criteria: ① random clinical trials; ② Aged 18-20. Exclusion criteria: ① Obviously not random clinical trials; ② Repeated study, syntheses, and Mata analysis articles. For the articles that we found, we conducted a search for each full-length article.DATA EXTRACTION: A total of 81 articles accorded with the standards were involved.DATA SYNTHESIS: From the comprehensive analysis of involved articles, we found that more college students' use the web just for communication, entertainment, excitement, and spiritual sustenance.CONCLUSION: This paper analyzes the modern college students' behavioral and psychological characteristics in Internet use from the perspectives of self-awareness, mood and emotion motivation, sexual psychology and so on respectively. It also puts forward an educational policy on how to instruct the college students to make use of the Internet properly and overcome the problems of psychology towards the Internet.

Objective:To analyze the failure factors in root canal therapy(RCT)by analysis of the failed cases.Methods:215 teeth of 206 patients were analyzed according to former treatment records,clinical examination,pretreatment radiographs,and root canal exploration during endodontic retreatment.Results:67.9% of failed teeth showed apical radiolucency,12.6% showed under-filled canal material without apical radiolucent area,and 8.4% involved coronal leakage.When examined with X ray,84.2% of teeth indicated under-filling,and most canals of premolars and molars had poor taper.Most missed root canals were found in maxillary first molars and premolars.18.1% of teeth had complications such as ledge and apical transportation.Conclusions:The factors associated with RCT failure involve cleaning,obturation and poor seal of root canal system resulted from inadequate shaping.Coronal leakage and missed canals also contribute to the endodontic treatment failure.

Background The reduced glutathione(GSH) is a water soluble antioxidant in retina and it is an important component of thiol.Diabetes induce the oxidative response of retina.It is very important to detect the content of thiol in retina for the evaluation of its antioxidant role.Objective This study was to investigate the effect of carnosine,aspirin and a combination of carnosine and aspirin eye drops on the change of the thiol contents in the retina of the diabetic rats. Methods Diabetic models were induced in 76 clean male Sprague-Dawley(SD) rats by intraperitoneal injection of streptozotocin(65 mg/kg).The 0.02 mi sodium citrate buffer was injected in 12 control rats.1%camosine eye drops or 0.05%aspirin eye drops were topically administered respectively in carnosine group (19 rats)or aspirin group(19 rats)two times per day for a period of 8 weeks,and the combination of 1%carnosine eye drops and 0.05%aspirin eye drops was used in the combination group at the same way(19 rats).At the end of 8 weeks.The animMs were sacrificed and the eyeballs were enucleated for the detect of thiols in the retina by colorimetry.Results The body weight of SD rats was significantly reduced and the blood glucose level was evidently arose in model group,camosine group and aspirin group in comparison with control group(P＜0.05),but those of combination group compared with control group was insignificantly changed(P＞0.05).The level of thiol in retina was(1 90.35+46.62)ms/gprot in model group,showing a considerable decrease in comparison with control (202.77+53.49 ms/gprot)(P＜0.05).However,the thiol level rose to(378.40±165.06)mg/gprot in camosine group and(351.88±211.62)mg/gprot in aspirin group,showing a statistically significant difference in comparison with control group(P＜0.05).The content of thiol in combination group compared with control group Was insignificant changed(166.972±50.287 mg/gprot us 202.77±53.49 mg/gprot).Curiously the levels in the combination group were not different from the normal control group. Conclusion The carnosine and aspirin eye drops may protect the retina against the oxidative damage caused by diabetes.