Objective To establish the retrovirus expression vector containing glucose transporter-2 ( GLUT-2) gene and its stable virus producing cell line,for the construction of new "artificial islet beta cells". Methods Plasmid pCB7 /GLUT2 was digested by EcoRⅠ/BamHⅠ and cloned to retroviral vector PLXSN for the construction of recombinant plasmid pL-GLUT2-SN,which was identified by enzyme digestion and sequencing,and then transfected to packaging cell line PA317. Titer of the recombinant virus was detected. Cell lines with a high titer were cloned and identified by PCR and RT-PCR. Results The retrovirus expression vector pL-GLUT2-SN was established,in which the insertion site and reading frame of target gene were confirmed by enzyme digestion and sequence analysis. The highest titer of virus producing cell line PA317 /GLUT2 was 7. 1 ? 105 CFU/mL. PCR and RT-PCR showed that the GLUT-2 gene was integrated into PA317 /GLUT2 and stably expressed. Conclusion A retrovirus expression vector containing GLUT-2 gene and its stable virus producing cell line PA317 /GLUT2 are successfully constructed,thus laying a foundation for the construction of "artificial islet beta cells".

Objective To observe the differences of the reactive oxygen species(ROS) and mitochondrial membrane potential(??m) in mitochondrial DNA-depleted SK-Hep1 and its parent cells.Methods ?0SK-Hep1 cell line was generated by treating SK-Hep1 with ethidium bromide.Flow cytometry(FCM) and laser scanning confocal microscopy were used to detect ROS and ??m.Results Fluorescence intensity of ROS in ?0SK-Hep1 cells was significantly higher than that of the parent cells(35.5 vs 15.6),and the ??m was lower in ?0SK-Hep1 cells(55.0 vs 65.9).Conclusion ?0SK-Hep1 cells showed elevated ROS and lower ??m.These changes might contribute to the variation of phenotypes.

Objective To explore the relationship between mitochondrial DNA microsatellite instability(mtMSI) and expression of Bcl 2 and Bax mRNA in gastric cancer and its precancerous lesions. Methods mtMSI and expressions of Bcl 2 and Bax mRNA were detected with PCR SSCP and RT PCR. Results Expressions of Bcl 2 mRNA in intestinal metaplasia (IM,53 3%) and dysplasia (Dys, 70%) were significantly higher than that in normal control(10%, P

Objective To explore the relationship between mitochondrial DNA deletion and malignant phenotypes of human lung cancer cells. Methods Two rho? derivatives of 95C and 95D were generated by treating the cultured cells with ethidium bromide. Agarose colony formation assays and Transwell invasion assays were carried out to detect the phenotypes of colony formation and invasiveness of the cultured cells, respectively. Cell growth was determined by MTT. Results The partially mtDNA-deleted cells exhibited stronger capacity of colony formation and invasiveness, and faster growth rates than their respective parental cell lines. Conclusion Mitochondrial DNA deletion might play a role in the formation of malignant phenotypes of human lung cancer.

Objective Mitochondrial DNA (mtDNA) may be the target of oxygen free radicals related to Helicobacter pylori (H.pylori) infection, partly because of lack of protective histones and partly because of inefficient DNA repair systems. In this study we investigated the relationship between mitochondrial microsatellite instability (mtMSI) as well as integration of mtDNA in the nuclei of gastric mucosa and H.pylori infection. Methods H.pylori was detected using PCR and modified Giemsa staining. The mtMSI and the sequences of mtDNA in the nuclei were detected by PCR SSCP and in situ hybridization. Results The mtMSI was detected in 11 of 30 (36.7%) cases of gastric cancers, 2 of 15(13.3%) of intestinal metaplasia, 2 of 10 of dysplasia and 1 of 10 of chronic atrophic gastritis. The integration of mtDNA in the nuclei was detected in 20.0% (6/30) of gastric cancer, 1/10 of dysplasia, 6.7%(1/15)of intestinal metaplasia and 1/10 of chronic atrophic gastritis. The frequencies of mtMSI and integration of mtDNA in H.pylori positive group (12/39 and 8/39) were each significantly higher than those in H.pylori negative group (4/36 and 1/36, P 0.05), mtMSI and integration of mtDNA in the nuclei tended to occur in gastric mucosa with cagA positive H.pylori infection(10/25,6/25), but less often to occur in gastric mucosa with cagA negative H.pylori infection(2/14, 2/14). Conclusions The mtMSI and integration of mtDNA may be involved in the carcinogenesis of gastric mucosa and H.pylori infection might contribute to the mtMSI and integration of mtDNA.

Objective To explore the feasibility ,effectiveness ,recurrence rate and complications ,etc .of endoscopic retrograde stenting in treating acute appendicitis .Methods A retrospective analysis was conducted on seven patients with the clinical diagnosis of acute appendicitis ,complying with the following treatment steps :(1) normal saline for high cleansing enema in 2-3 times ;(2) en-doscopically douching ileocecal junction and observing the opening of appendix ;(3) placing the guide wire into the opening of appen-dix under the guidance of imaging tube ,and then conducting angiography imaging ;(4) repeatedly douching with metronidazole;(5) implant the appendix stent ,draining the inflammatory secretions out and then douching ;(6 ) observing postoperative abdominal pain ,fever ,bowel and other conditions;(7) reviewing with the enteroscopy and removing the stent 2 weeks later .At 2 before the surgery and 48 after it ,all patients were administrated with antibiotics for anti-infective treatment .The follow-up was made from 8-15 months after the surgery .Results Among these 7 patients ,4 patients had successful appendix stenting :the abdominal pain sig-nificantly alleviated after the surgery ;the proportional level of white blood cells(WBC)recovered during 24-48 after the surgery .15 after discharge ,two patients returned to hospital and their appendix stent removal was successful;during the operation ,smooth mu-cosa at the opening of appendix was observed .The stents of two patients spontaneously fell off ,and normal morphology of the ap-pendix opening was observed at review .During the postoperative follow-up of 8-15 months ,one patient relapsed and underwent sur-gical treatment in the general surgery department .The other three patients did not undergo appendix stenting due to the unsuccess-ful intubation .Conclusion The treatment of acute appendicitis with endoscopic stenting has the advantages of little trauma ,high safety and significant efficacy .However ,this method still requires large-scale and multicenter randomized controlled clinical trials to evaluate its feasibility and long-term efficacy .

Ascites is the most frequent decompensating event of cirrhosis.At present,ascites recurs at a high rate due to lack of effective management strategy and is frequently complicated with spontaneous bacterial peritonitis,hepatorenal syndrome,and liver failure,which increase the fatality rate.Albumin treatment for hepatocirrhosic ascites has a long history,but it is limited as an acute or short-term treatment.In contrast,long-term albumin administration represents a completely different treatment paradigm.Results from several recent clinical studies indicate that long-term albumin treatment can be able to modify the disease courses of some decompensated cirrhosis when albumin is given at a sufficient dose for a sufficient time.In this review,we analyze the available data acquired from long-term albumin treatments,trying to establish a secure and effective management scheme involving maximal target popula-tion,albumin dose,administration time,and standards for albumin withdrawal,and thus provide references for the clinical practice.

BACKGROUNDTo isolate and identify the genes related to cancer metastasis by comparison of two cell strains with different metastasis potentials subcloned from human lung giant cell carcinoma cell line.

METHODSSuppression subtractive hybridization (SSH) was used to compare the levels of gene expression between the two cell strains and SSH library was constructed. After screening the library by gene chip, the expressed sequence tags (ESTs) with different expressing level were sequenced and blasted with GenBank.

RESULTSSeventy-nine genes were obtained that were expressed much higher in PLA-801D than in PLA-801C, including two full-length cDNA. GenBank Accession numbers of the two cDNA, named MAG-1 and MAG-2, were BC006236 and BC002420, the 8.5 kb MAG-1 gene was composed of four exons and located on the chromosome of 4q21. The MAG-2 gene, which was made up by 9 exons, had a length of 5.2 kb and its location was 2q35. Both sequences had open reading frames (ORF) and promoters before the theoretical transcription start points. Using special software, the secondary structure of theoretical products of the two cDNAs was prognosticated, α-helix was the main proportion, but β-pleated sheet and random coil were also included.

CONCLUSIONSThe expression of MAG-1 and MAG-2 has significant differences in these two cell strains, so they might impact tumor metastasis in some ways that are still uncharted.