BACKGROUND: Construction of lentivirus vector has the advantages of simplicity,it is regard as the most effective and successful method in transgene therapy. OBJECTIVE: To clone the metastasis suppressor gene KiSS-1 from human normal placenta tissue and construct its lentivirus vector. DESIGN,TIME AND SETTING: The open experiment was performed at the Laboratory of Fujian Normal University from September 2006 to December 2007. MATERIAL: The pNL-IRES2-EGFP vector was conservated by the Laboratory of Fujian Normal University. METHODS: Total RNA was extracted from human placenta tissue. The opening reading frame cDNA of KiSS-1 was isolated by using RT-PCR,and cloned into its lentiviral vector pNL-IRES2-EGFP to construct expression plasmid pNL-IRES2-EGFP-KiSS-1. MAIN OUTCOME MEASURES: Clone objective gene fragment of KiSS-1,restriction enzyme digestion and gene sequencing of the recombinant plasmid pNL-IRES2-EGFP-KiSS-1 were observed in the study. RESULTS: The nucleotide sequence isolated from the recombinant plasmid pNL-IRES2-EGFP-KiSS-1 was confirmed the same as expected by restriction enzyme digestion and gene sequencing. CONCLUSION: The recombinant plasmid pNL-IRES2-EGFP-KiSS-1 has been constructed successfully.

BACKGROUND: Traditionally, bee venom can treat rheumatic arthritis,rheumatoid arthritis(RA) and so on, but it has strong side effects. So it has been hoped for a long time that the effective angle component could be screened from bee venom, which can be used for the treatment of arthritis perfectly than bee venom.OBJECTIVE: To investigate whether bioactive peptide from bee venom could inhibit infection of arthritis by regulating immunological function so as to probe into a new treatment for RADESIGN: Completely randomized controlled experimental trial based on experimental animalsMETHODS: A municipal key laboratory of animal biology.MATERIALS: The experiment was carried out in the Chongqing Key Laboratory of animal biology from January 2001 to May 2002. Totally 80 rats of clean grade aged 2 to 3 months old with the body mass of 180 to 200 g were provided by Animal Experiment Center of Third Military Medical University of Chinese PLA. The experimental animal certification number was SYXK1 (army) 2002 -007. The animals were divided into 3 groups: normal control group( 10 cases), arthritis group( 10 cases), bioactive peptide group(30 cases).METHODS: Adjuvant-induced arthritis animal models were used and bioactive peptide were given to the animals by muscle injection to observe the knuckle volume and knuckle index changes.MAIN OUTCOME MEASURES: The effect of bioactive peptide from bee venom on the change of knuckle volume and knuckle index in adjuvant-induced arthritis ratsRESULTS: Ten days after injection of 0. 15 mg for each rat, the volume of the paw was (4.72 ±0. 58) mL and the knuckle index was (4.47 ±0.46) mL,which there was significant difference compared with the control group (P＜ 0. 05).CONCLUSION: P-peptide possibly has certain inhibitory effect on the development of the adjuvant-induced arthritis in Wistar rat, and will possibly be a potential therapeutic drug.

Objective To evaluate antitumor effect of adenovirus-mediated cytosine deaminase (CD) gene and 5-fluorocytosine (5-FC) on rectal cancer cells.MethodsCD gene was transinfected into HR-8348 rectal cacer cell lines with recombinant adenovirus mediation. The transinfection rate and expression of CD gene were detected. Plating efficiency and MTT method were used to examine inhibition of HR-8348 cell growth, and antitumor effect of CD/5-FC were evaluated on HR-8348 experimental tumor in nude mice.ResultsHigh transinfection effeciency and expression of CD gene were achieved in HR-8348 with adenovirus mediation. CD/5-FC system showed a remarkble effect of inhibition on plating efficiency and growth of the tumor cells transinfected with CD gene, but no act on tumor cells without CD gene transinfection. In a mixture of HR-8348 tumor cells with and without CD gene transinfection, CD/5-FC killed both types of tumor cells and revealed a powerful "by-stander effect". With nude mice experiment, 71.5% of inhibition of solid tumoral xenografts of HR-8348 cells was found.ConclutionCD/5-FC has significant antitumor effect on rectal cancer cells with adenovirus-mediated CD gene transinfection and has also "by-stander effect" on tumor cells.

Objective To study the expression of cathepsin B (CatB) and metallothionein (MT) in colorectal cancer tissue, and explore their association with lymphatic and hepatic metastasis. Methods Immunohistochemistry technique was used to assay the expression of CatB and MT in 82 cases of primary colorectal cancer, normal colon mucosa, and lymph node and hepatic metastasis. The relationship between the expressions of CatB and MT and the clinical stage or pathological grading of the colorectal cancer were analyzed. Results The rate of CatB expression in primary tumor, normal colon mucosa, and lymph node with metastasis and hepatic metastasis was 48.8%, 20.7%, 66.7%, and 59.1%, respectively, and that of MT expression was 53.7%, 26.8%, 71.4% and 72.7%, respectively. The rates of CatB and MT expression in primary tumor, and lymph node and hepatic metastasis were higher than that in normal mucosa tissue. The rate of coexpression of CatB and MT in lymph node and hepatic metastasis was higher than that in primary tumor and lymph node without metastasis. As regard to clinical stages, the positive rates of CatB and MT in Dukes C and D were higher than that in Dukes A and B (?2=11.024 4, 10.933 8, P

Objective To explore the influence of cathepsin B (CatB) expression on the ability of adhesion and invasiveness of colorectal cancer cells. Methods Human colon cancer cell line LoVo and rectal cancer cell line HR-8348 were used in the present study. The total length of CatB mRNA was cloned and transfected into the cells, and the CatB expression was detected by RT-PCR. Cell adhesion was detected with Matrigel and invasiveness was assessed with transwell method. Results CatB mRNA expression was detected and enhanced expression was found in LoVo and HR8348 cells tranfected with pcDNA3CatB. Cell adhesion rates to Matrigel stromatin in control LoVo cells, empty vector transfected cells and CatB transfeted cells were 0.461 6?0.148 7, 0.412 1?0.215 8 and 0.691 6?0.150 8, respectively. The adhesive rate of CatB transfected LoVo cells was higher than that of the control cells and the empty vector transfected cells with significant differences (F=5.839 0, P

Perinatal cardiomyopathy is a kind of idiopathic cardiomyopathy ,its morbidity rate is low but it may re‐sult in death . The present article made a review on etiological researches of perinatal cardiomyopathy of recent years ,aiming at providing assistance for clinical diagnosis and therapy .

Objective To study the differentially expressed proteins and their biological behavior in colorectal carcinoma tissues and the normal colonic mucosa by proteomics and molecular biology techniques. Methods The technique of fluorescence two dimension differential gel electrophoresis (2-D DIGE) was used to analyze the expression of differential proteins in normal colorectal mucosa and primary cancer foci. Liquid chromatography with electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS) was used to identify the differential proteins. Transfection experiment of colorectal cancer cells was performed with the differential protein cDNA, and the changes in cytobiological behavior were observed. Results Significant differences of protein expression levels were found by two-dimension electrophoresis. Eight differential protein spots were analyzed and identified. Human carbonic anhydrase Ⅱ and protein disulfide isomerase were detected in normal colorectal mucosa, but not in primary cancer foci. Phosphoglycerate kinase 1, fumarate hydratase and aldolase A were expressed in primary cancer. After transfection with human carbonic anhydrase Ⅱ cDNA, the abilities of Lovo cells were obviously reduced in invasiveness, chemotaxy motor and drug resistance. Conclusions Differences on protein expression levels are found between normal colorectal mucosa and primary cancer foci by 2-DE DIGE. The pathogenesis of colorectal carcinoma is related to the reduced expressions of carbonic anhydrase II and protein disulfide isomerase and enhanced expression of aldolase A. The technique of differential proteomics is useful in reaching a indepth understanding of the pathogenesis mechanisms of human colorectal cancer.

Objective To evaluate the target killing effect and metastasis prevention effect of soluble Fas combined with PKC inhibitor on the growth of human colorectal carcinoma implant in nude mice. Methods Orthotopic implantation and metastasis model of human colorectal carcinoma was reproduced in nude mice. Tumor tissue of tumor cell line HR-8348 with positive expression of FasL was implanted to the colonic wall of nude mice. After one week of tumor growth, mice were randomly divided into four groups according to the different agents injected into the peritoneal cavity. Twelve mice were in each group. The mice in the combined treatment group (recombinant soluble Fas coupled with PKC inhibitor + 5-Fu) were injected intraperitoneally 100?l (3mg/ml) recombinant soluble Fas coupled with PKC inhibitor and 0.5 mg of 5-Fu. (On the day of 0, 4, 8, 12 and16). At the same time, a group of tumor bearing mice were given recombinant soluble Fas coupled with PKC inhibitor only, and another group with 5-Fu only, and in the control group only normal saline was given. One month after implantation, tumor weight, inhibition rate and the presence of metastasis were evaluated respectively after the mice were sacrificed. Results Compared with control group, the orthotopically implanted tumors were significantly reduced in weight in mice treated with 5-Fu, recombinant soluble Fas coupled with the PKC inhibitor, and combined treatment, with respective inhibited rates of 43.1%, 79.9%, and 86.3%. Liver metastasis was also inhibited with significant decrease in incidence in 5-Fu group, recombinant soluble Fas coupled with the PKC inhibitor, and combined group compared with that in control group (75.0% vs 36.4%, 16.7%, and 0%). The incidence of peritoneal metastasis was also decreased significantly in 5-Fu, recombinant soluble Fas coupled with PKC inhibitor, and combined treatment compared with that in control group (100% vs 45.5%, 16.7%, and 8.3%, P

Objective To compare the clinical pathological characteristics in young and old patients with colorectal cancer (CRC) and investigate their relationship with prognosis. Methods A retrospective review was made in 68 CRC patients less than 35 years old and 322 CRC patients older than 65 treated in our hospital from July 1993 to July 2003. Their clinical manifestation, pathological feature, Dukes staging, misdiagnosis rate and results of following-up were compared. Results The main manifestation in young group was abdominal pain (69.1%), but in old group was hemafecia or mucous bloody stools (53.7%). The ratio of poorly differentiated neoplasm was obviously higher in young group (48.5%) than in old group (20.5%) (P

BACKGROUND: Recently, miR-378 has been shown to modulate the anti-hypoxia capacity of bone marrowmesenchymal stem cells (BMSCs) and reduce cell apoptosis under hypoxic conditions.OBJECTIVE: To investigate the benefits of miR-378-upregulated BMSC transplantation in a rat model of acutemyocardial infarction.METHODS: Primary rat BMSCs were cultured in vitro. Until passage 3, the cells were infected with the lentiviruscarrying synthetic miR-378 gene fragments. A rat model for acute myocardial infarction was constructed by ligatingthe left anterior descending artery. Thereafter, the animals were randomly assigned to three groups: control group(n=10), BMSCnull group (n=16) and BMSCmiR-378 group (n=16). In the latter two groups, 50 μL of normal salinecontaining 1×107 empty virus-transfected or miR-378-transfected BMSCs was injected into the region of myocardialinfarction, respectively. Only 50 μL of normal saline was injected in the control group. Twenty-four hours later, theapoptosis of transplanted BMSCs was evaluated with TUNEL, and expression level of vascular endothelial growthfactor and transforming growth factor-β was detected using western blot assay. Four weeks after treatment, the leftventricular function of rats was assessed by echocardiography, and then histological and molecular biology analyseswere performed.RESULTS AND CONCLUSION: At 24 hours postoperatively, there were less apoptotic BMSCs and higher expressionlevels of vascular endothelial growth factor and transforming growth factor-β in the BMSCmiR-378 group than in theBMSCnull group (n=6, P < 0.001). Four weeks later, there were more transplanted BMSCs and BMSCs-derivedcardiomyocytes in the BMSCmiR-378 group than the BMSCnull group (n=10, P < 0.001). Moreover, increased new vesseldensity (P < 0.001), decreased infarcted area (P < 0.001), preserved left ventricular ejection fraction (P < 0.05), reducedleft ventricular end-diastolic volume (P < 0.05) were found in the BMSCmiR-378 group, compared with the other two groups.The above parameters were better in the BMSCnull group than the control group (P < 0.05). Overall, the upregulation ofmiR-378 could enhance the capability of BMSCs against hypoxia, and consequently promote myocardial repair afterimplantation, providing a new strategy for cell therapy of myocardial infarction.