The whole length cDNA of human interferon-?(IFN-?) gene including signal peptide was constructed to retroviral vector pLXSN and then packaged with PA317. A Human Hepetoma Cell Line(HHCL) and a human gastric cancer cell line (MKN-45) were infected by using supernatant containing retroviral vector with IFN-?. the gene-modified tumor cell. were isolated by G418 selection. Some changes were found in cell cycle, expression of HLA class Ⅰ and classⅡ, and tumorigenecity. Elevated immunogenecity and abrogated rumerigenecity suggest a means for generating gene-modified tumor cell vaccine. for the treatment of cancer.

Retroviral vector was used to introduce the interleukin - 2 (IL -2) gene into H22 cells, a BALB/c mouse hepatoma cell line. The IL-2 gene and marker gene (NeoR) were assessed by using PCR and RT-PCR methods. FACS analysis demonstrated that the cells with low DNA content in IL-2 gene modified H22 cells were more than these in control H22 cells. Electron microscopic morphological structure showed that some cells in IL-2 gene modified H22 went into apoptotic cell death. The study demonstrated that apoptotic cell death of H22 - IL-2 cells was induced by the IL-2 gene modification. It may be one of mechanisms of decreased tumorigenicity of IL - 2 gene modified tumor cells.

We compared the ability of tumorigenicity of TNF-? gene-modified and unmodified H22 tumor cells, and therapeutic functions of the irradiated cells. Results indicated that H22-TNF-? cells were less tumorigenic as compared to H22 and H22-LXSN cells. In case of treatment groups, injected with irradiated tumor vaccine in 3 or 6 days after inoculation of H22 cells, the tumor growth was suppressed.

Objective To investigate the changes of interleukin-21 (IL 21) and interleukin-21 receptor (IL 21R) expression level in Crohn's disease (CD) patients before and after accepted infliximab (IFX) treatment.Methods From June 2009 to July 2011,twenty-two CD patients met the research criteria were recruited at Tenth People's Hospital of Tongji University.Patients were treated with infliximab at weeks 0,2,6,and 16 healthy individuals were set as healthy control group at same time.Peripheral blood of healthy control group was taken at regular physical examination and blood of CD patients was taken before treatment and 10 weeks after treatment,intestinal mucosa biopsy samples were taken under colon endoscopy examination.The changes of Crohn's disease activity index (CDAI),erythrocyte sedimentation rate (ESR),C-reactive protein (CRP) in CD patients were observed.The change of IL-21R in peripheral blood CD4+ T lymphocytes was detected by flow cytometry.The change of IL-21 expression at mRNA level in intestinal mucosa was determined by realtime quantitative polymerase chain reaction (PCR).The data were analyzed by t test.Results Before treatment,the level of IL21R in peripheral blood CD4+ T lymphocytes of CD patients (12.25％±3.25％) and the expression of IL-21 at mRNA level in inflamed intestinal mucosa (1.38±0.32) were both significantly higher than those of healthy controls (4.25 ％ ± 1.41％,0.44±0.18),the differences were statistically significant (F=15.88,6.75 ; both P＜0.05).At 10th week,the level of IL-21R in peripheral blood CD4+ T lymphocytes of CD patients (8.12％ ± 2.05％) and the expression of IL-21 at mRNA level in intestinal mucosa (0.77 ± 0.24) were both significantly lower than those before treatment,the differences were statistically significant ( t=4.880,8.019; both P＜0.01).Before treatment,ESR,CRP and CDAI of CD patients was (46.8±11.4) mm/1 h,(52.4±11.5) mg/L and 319±74,which was (23.5±9.0) mm/1 h,(11.6±4.6) mg/L and 113±42 after treatment,the differences were statistically significant (t=9.485,16.458,11.100; all P＜0.05).Conclusion The IL-21 expression of active CD patients decreases after IFX treatment,which indicates that IL-21 may involve in IFX induced clinical remission of active CD.

Drugs, Chinese Herbal ; Humans ; Rhabdomyolysis

Objective To construct recombinant adenovirns containing small interfering RNA (siRNA) targeting human VEGF-C mRNA,then to study the inhibitory effects of adenovirus-mediated VEGF-C siRNA on growth and lymphangiogenesis of human colon cancer in BABL/c nude mice model.Methods In vitro:the specific siRNA sequence targeting human VEGF-C mRNA was selected.The homologous double-strand DNA was designed and synthesized.After such DNA was inserted into pDC316-EGFP-U6 by BamHI and HindⅢ,pDC316-VEGF-C siRNA-EGFP-U6 was obtained,then it was co-transfected into 293 cells with the hone plasmid pBHGF35.After generated by homologous recombination,Ad5F35-VEGF-C siRNA-EGFP-U6 was obtained,and it was packaged and amplified in 293 cells.The human colon cancer model was established in nude mice by hypo-injection LoVo cells.They were divided randomly into four groups (n=6):adenovirus,virus without target gene,single siRNA and PBS control group.Injecting intervention intra-tumorly in each groups,calculating the tumors volume and drawing the growth curve,calculating micro-vascular density (MVD) and micro-lymphatic density (LVD) by staining respectively of the lymphatic and vascular with anti-LYVE-1 monoclonal antibody and anti-CD<,34> monoclonal antibody were completed.Results Adenovirus group tumor sizes were smaller than other groups.The LVD were 8.47±2.1 and 17.35±4.7 (P<0.05) in adenovirns group and the PBS control group.The MVD were 22.65±6.04 and 23.19±7.63 (P>0.05) in adenovims group and the PBS control group respectively.Conclusion The adenovirus-mediated VEGF-C siRNA can significantly inhibit the growth and lymphangiogenesis of human colon cancer in nude mice model,hut have no effect on microangium vessels growth.

Objective: In China, doctors at TCM hospitals and clinics often divide patients with a Western medicine (WM) disease into several syndrome classes from the TCM perspective and treat patients in different classes using different principles. A key problem is how to carry out the classification properly. We propose an evidence-based ap-proach for solving the problem where evidence is obtained by analyzing unlabeled symptom data using latent tree models.Method: In previous work, we have shown how latent tree analysis of symptom data can be used to identify TCM syndrome classes among patients with a WM disease. In the paper, we investigate how to establish classification rules for distinguishing between the classes.Results: We have applied the method to a data set about Vascular Mild Cognitive Impairment that involves 93 symptoms and 803 patients. Nine syndrome types are identified, along with the corresponding classification rules. Conclusions: An evidence-based approach to the TCM patient classification prob-lem has been developed. The approach can be used to answer the following questions about a WM disease: What TCM syndrome classes are there? What are the sizes of the classes? What are the statistical characteristics of each class? How can one differentiate between the different classes?

Objective To create Mucin gene 1 (MUC1) antisense peptide nucleic acid (PNA),and to observe its effects on MKN-45 cell invasion and explore the mechanism. Methods The sequence of antisense PNA was designed according to MUC1 gene sequence and transfected into human gastric cancer cells (MKN-45) by liposome,and the empty vector group (randomized control group)and blank control group (negative control group) were involved. The expression of MUC1 was detected by real time quantitative PCR and the changes of E-cadherin expression were also observed.The effects on gastric cancer cell invasion were tested with transwell chamber assays.Results The expression of MUC1 gene was effectively suppressed by the 3 created antisense PNA,and their expression level (0.62±0.18,0.49±0.12 and 0.60±0.21) was significantly lower than that of negative control group (1.18 ± 0.03,P ＜ 0.01). There was no significant difference between radomized control group and negative control group (1.00±0.04,P=0.657).After MUC1 PNA transfected,the capability of gastric cancer cell invasion decreased significantly (P=0.005).And the expression of E-cadherin at mRNA and protein level was up-regulated.Conclusions There is negative correlation between MUC1 and E-cadherin expression in gastric cancer cell MKN-45.The capability of tumor cell invasion is significantly inhibited by suppressing MUC1 gene expression.

Objective To investigate the expression of microRNA (miRNA)-10a in the intestinal mucosa,serum and peripheral blood mononuclear cell (PBMC) of patients with inflammatory bowel disease (IBD) and explore its role and relevance in the pathogenesis of the disease.Methods The intestinal or colonic mucosal biopsy specimens of nine active ulcerative colitis (UC) patients,11 active Crohn's disease (CD) patients and eight patients with negative colonoscopy result as control were collected.The sera of 12 active UC patients,13 active CD patients and nine healthy controls were collected.The PBMC of nine active UC patients,11 active CD patients and eight healthy controls were collected.The expression of miRNA-10a in the intestinal mucosa,sera and PBMC and the expression of IL-12/IL-23 p40 in the intestinal mucosa were detected by real-time polymerase chain reaction (PCR).Each 8 cases of active UC and CD patients were collected.The intestinal mucosa before infliximab (IFX) treatment and six weeks after three times of IFX treatment were collected.And at same time,the intestinal mucosa of 11 active UC patients and 10 active CD patients were collected and cultured for 18 hours stimulated with IFX in vitro and then the expression of miRNA-10a in the intestinal mucosa was tested.One-way analysis of variance was used for comparison in three samples.Paired t-test was used for two samples comparison.Spearman test was used for correlation analysis.Results Compared with healthy controls,the expression of miRNA-10a in the intestinal mucosa,serum and PBMC of UC and CD patients significantly decreased (F=38.45,30.46 and 14.74,all P＜0.05).There was no statistic significance between UC and CD groups.The expression of IL-12/IL-23 p40 in the intestinal mucosa of UC and CD patients significantly increased (F=32.90,P＜0.05).The expression of IL-12/IL-23 p40 was negatively correlated with the expression of miRNA-10a in the intestinal mucosa of CD patients.After three times of IFX treatment,the expression of miR-10a in the intestinal mucosa of IBD patients significantly increased (t=3.341,3.382,both P＜0.05).After stimulated with IFX in vitro,the expression of miRNA-10a in the intestinal mucosa significantly increased (t=3.095,7.193,both P＜0.05).Conclusions miRNA-10a was closely correlated with the inflammation of IBD patients and with the role of targeting IL-12/IL-23 p40.miRNA-10a might be a new target for the IBD treatment.

Objective To analyze the feasibility and practicality of the navigational template for antegrade lag screw fixation of the fractures in the posterior column of acetabulum.Methods Spiral CT scan data of 40 normal adult pelvis was collected.3D CT scans of pelvics were performed.Virtual 3D pelvic models were reconstructed with software Mimics 10.01.Virtual cylindrical implants were placed along the longitudinal axis of the acetabular posterior column via the ischial tuberosity among the ipsilateral hemipelvis and some anatomical parameters were measured.To adjust cylinder position to determine the best projection point,the shortest distance of the point to the linea terminalis and the anterior border of the auricular surface,the angle between the cylindrical and the coronal plane (α),and the angle between the quadrilateral district and iliac fossa plane (β) were measured respectively.The 3D models were imported into software UG 6.0.The models of navigational templates were designed according to the anatomic features of the acetabulums and the measured parameters.16 cases of dry left side of pelvis specimens were collected.Simulate acetabular posterior column screw fixation from different screw holes and verify the accuracy of navigation template assisted in the nail.Results The navigational templates was designed and manufactured successfully which was more consistent with the anatomical features of the quadrilateral plane.Placement of lag screw from the first hole:6.3％ achieved accurate positioning,75％ well positioning,and 18.7％ loose.From the second hole:81.3％ had accurate positioning,18.7％ well positioning.From the third hole:37.5％ had accurate positioning,62.5％ loose.Conclusion The navigational template designed according to the anatomic features of acetabulum and the measured parameters can accurately assist lag screw placement.