AIM:So far,endostatin is found playing the strongest effect to inhibit endothelial cells.Because endostatin protein is instable,it is difficult to prepare and apply.Gene therapy is used to assist endostatin to inhibit angiogenesis.This study constructed recombinant adenovirus of human endostatin gene using AdEasy-1 system to investigate the feasibility of expressing endostatin protein in ECV304 cells(human umbilical vein endothelial cells).METHODS:The experiment was performed at College of Life Science,Sun Yat-set University from March to October 2006.Endostatin was amplified from Pshuttle-Endostatin plasmid with PCR and subcloned into the pAdTrack-CMV shuttle vector after sequencing and identification.The resultant plasmid was co-transduced into E.coli BJ 5183 cells with pAdEasy-1 plasmid for homologous recombination.The recombinant plasmid was detected by restriction analysis and DNA sequence analysis.The linearized recombinant plasmid was transfected into AAV 293 cells with LipofectAMINE2000.The recombinant adenovirus was detected by GFP,PCR and restriction analysis.The virus was used to infect ECV304,and the expression of endostatin in vitro was detected by the techniques of RT-PCR and Western blot.RESULTS:The positive clones of the recombinants were constructed and confirmed by restriction endonuclease analysis and DNA sequence analysis.AAV293 cells were transfected.The titer of viral liquid was 2.06?1010 pfu/mL.The transcription of endostatin mRNA in ECV304 cells was checked by RT-PCR and endostatin protein could be detected in the culture by Western blot analysis.CONCLUSION:The recombinant adenoviral vector pAd-Endo carrying human endostatin is successfully constructed,and effective expression is found in ECV304 cells in vitro.

Objective To discuss progress in Qinghai Tibetan Thymidylate synthase enhancer in patients with gastric cancer (TSER),ERCC1 polymorphism of genes,gene expression and mRNA studies.Methods 210 cases of advanced gastric cancer were enrolled as required to complete the treatment and follow-up.TSER,ERCC 1 gene polymorphism were tested using PCR-RFLP method.Analysis of gastric cancer is based on the results of gene polymorphism and clinical pathological factors in relations.The expression of ERCC1 mRNA was detected by RTPCR,and tracking access after treatment.Analyzing the relationship among ERCC1 mRNA expression,clinical treatment of overall survival,total response rates and gene polymorphism.Results In patients with advanced gastric C/C in Qinghai ERCC1 gene frequency was 54.2％,C/T frequency was 42.7％,T/T ERCC1 gene frequency was 5.6％.ERCC1 mRNA expression and clinical effect in the treatment of pathological factors were not related,ERCC1 mRNA and gene ERCC1 polymorphism had no relevance.Conclusions Progress in Qinghai Tibetan TSER gene is highly expressed type 3R/3R in patients with gastric cancer.ERCC 1 gene polymorphisms with type C/ C is overexpressed.ERCC1,TSER gene polymorphism and clinicopathologic parameters in advanced gastric is not related.Extension of Tibetan ERCC1 mRNA in gastric cancer patients expression has no association with ERCC1 gene polymorphism.

ObjectiveTo explore the characteristics and the pathogenetic mechanism of prospective memory in patients with chronic primary insomnia. Methods Forty-two patients with chronic primary insomnia and forty age,gender,education,and cultural level matched healthy adults were assessed with a battery of neuropsychological tests including event-based prospective memory (EBPM)and time-based prospective memory (TBPM) tasks.ResultsCompared with the healthy controls ( EBPM: 5.58 ± 0.35 ;TBPM: 4.10 ± 0.32),the patients with chronic primary insomnia had a significant loss in EBPM (3.44 ±0.40,t =3.933,P =0.000),but a insignificant impairment in TBPM (3.76 ± 0.29,t =0.787,P =0.433 ).The retrospective component of event-based prospective memory between the two groups had no significant difference ( t =1.308,P =0.195 ) ,neither did the retrospective component of time-based prospective memory ( Z =- 0.776,P =0.437 ).ConclusionThese results suggest that the patients with chronic primary insomnia have impairment in EBPM,but not in TBPM and the retrospective component of prospective memory.

A liver transplant invol ves more medical aspects than complex surgery,with different characteristics than general surgery.The medical risks the patient disputes focus on the informed consent rights.Analysis of dispute causes,especially through the relevant study of jurisprudence,suggests that we should be highly sophisticated with informed consent rights.to prevent disputes and improve the quality of clinical diagnosis and treatment.

In this paper, the theory of electrical character and adsorption of solidliquid interface was applied to study the preparation process for borneol-mi-croencapsule with co-coacervation using gelatin and arabic gum as materials. The optimum process conditions for preparing microencapsule was gained. This study method provided a reliable basis for raising the inclusion rate, decreasing the capsule materials in quantity and drawing up the process conditions.

Objective To approach the directive function of time efficiency of first-aid to war wound and trauma. Methods To analyse the relationship between time and war wound or trauma death based on the statistical data of those wounds, then to define the time window of first aid aiming at the peak value of death. Results Bleeding and asphyxia should be virtually controlled in “emergency platinum 10 minutes”, resuscitation from shock should be within 30 minutes, at the same time, a definite operation to save life should be performed in “golden an hour”. Conclusions It is important to raise the time efficiency in each stage by training and improving the military affairs. To spread the first-aid knowledge may decrease cripples and mortality.

Medical nutrition therapy (MNT)is the basic and long-term treatment of diabetes patients. At present,the dietary structure of type 2 diabetes mellitus(T2DM)patients in China has yet to be further improved. This article focused on the effect,dietary structure,food selection and guarantee of compliance of MNT in T2DM.We recommended that the dietary pattern should be designed based on the traditional dietary habits around the country,combined with people living and working reality. The rational utilization of network platform can improve the efficiency and compliance of nutrition education.

Objective: To compare the differences in gene expression between murine intestine intraepithelial lymphocytes (iIELs) and splenic T lymphocytes, establishing the cDNA subtractive library of iIELs, analyzing the iIELs special genes. Methods: The iIELs and splenic T lymphocytes were isolated by density gradient in Percoll and nylon column respectively. cDNA subtractive library of iIELs was established by improved subtractive hybridization. The cDNA was ligated with a different adaptor. After 2 rounds of hybridization and suppression PCR amplification, the PCR products were directly inserted into the plasmid vectors. The ESTs special for iIELs were screened by reverse Northern blot. Results: Fifty ESTs with obvious difference in the cDNA subtractive library of iIELs were screened. These ESTs were squenced and submitted to Genebank. The ESTs were analyzed through BLAST, and 14 genes special for iIELs were obtained. These genes were associated with the pattern recognition of pathogen, cytokine signal transduction and proteinase inhibitors. Conclusion: iIELs are important components of intestine natural immunity, it can recognize the conservative antigens of bacteria and can be affected by cytokines and proteinase inhibitors, leading to unique cellular behavior--more resting and less activation.