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.

Drugs, Chinese Herbal ; Humans ; Rhabdomyolysis

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.

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 elucidate the correlation between the single nucleotide polymorphism of CKLF-like MARVEL transmembrane member 5 ( CMTM5 ) gene rs723840 and the occurrence of high on aspirin platelet reactivity ( HAPR) . Methods:The present study is a case-control study. A total of 210 hospitalized patients in Peking University First Hospital were enrolled. Aspirin response was assessed by 0. 5 g/L arachidonic acid (AA)-induced platelet aggregation ratio (PR), and ≥3/4 quartile of PR of the population was defined as HAPR. Accordingly all the enrolled 210 coronary artery diseases ( CAD) patients were divided into HAPR group and No-HAPR group. The genotypes were determined by poly-merase chain reaction ( PCR) and sequencing analysis for rs723840 of CMTM5 gene. Results:The geno-type frequencies in rs723840 C>T of CMTM5 gene conformed well to the Hardy-Weinberg equilibrium in both HAPR group and No-HAPR group. Between the two groups, the genotypes frequencies in HAPR and No-HAPR groups were 48 . 4%, 51 . 6%, 0 . 0% and 73 . 7%, 22 . 9%, 0 . 034%, respectively ( P=0. 004). The C, T allele frequencies were significantly different in the two groups (P =0. 031,OR =0 . 501 , 95%CI:0 . 264-0 . 947 ) . Conclusion:Our study finds a significant correlation between CMTM5 gene rs723840 polymorphism and high on aspirin platelet reactivity.

Objective:To elucidate the correlation between urinary 11-dehydro-thromboxane B2 ( 11 dhTxB2 ) and clinical efficacy of aspirin treatment in patients with type 2 diabete and coronary artery disease ( CAD) . Methods:In this prospective cohort study, 169 aged patients with type 2 diabete accom-panying CAD in Peking University First Hospital were enrolled. The level of urinary 11dhTxB2 was detec-ted using enzyme-linked immuno-sorbent assay. Low aspirin response or high on aspirin platelet reactivity (HAPR) was defined as urinary 11dhTxB2>1 500 ng/g. All the included patients were divided into two groups based on the results, HAPR group and No-HAPR group. Results:Baseline urinary 11dhTxB2 of the patients with type 2 diabete accompanying CAD was ( 3 687 ± 3 052 ) ng/g, while the urinary 11dhTxB2 was (1 954 ± 859) ng/g in patients after 100 mg/d aspirin treatment (P<0. 001). Preva-lence of HAPR in patients with type 2 diabete accompanying CAD were 32 . 5%. Within a mean follow-up time of 12 months, the outcomes occurred more frequently in HAPR group than in No-HAPR group ( P<0 . 05 ) . Conclusion:Urinary 11 dhTxB2 can be recognized as an effective indicator in evaluating aspirin clinical efficacy of patients with type 2 diabete accompanying CAD.

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.