Objective To observe the effeet of cotransduction of wt-p53,B7-1 and GM-CSF genes mediated by recombinant adenovirus on cell growth, apoptosis and immunogenicity of neuroblastoma cell line SH-SY5Y, Methods We transfected SH-SYSY cells separately with recombinant adenovirus, recombinant adenovirus mediating human wild type p53(wt-p53) gene and BB-102.We separately named them with SH/Ad,SH/p53 and SH/BB-102 group.The protein expression of p53, B7-1 and GM-CSF were measured by Western blot, FCM and enzyme linked immunosorbent assay. Cell number was counted and growth curves were drawed. Cell apoptosis was tested by FCM. Lymphocyte proliferation and cytokine secretory was evaluated by mixed lymphocyte culturing. Results After transection with BB- 102, p53, B7-1, and GM-CSF, high-efficiency expression of target genes were found. Growth of the cells was inhibited, apoptosis was induced in the SH/p53 and SH/BB-102 groups. Proliferation of lymphocytes was stimulated and cytokine increased visorously in the SH/BB-102 group. Condusions After transfection with BB-102, growth of the SH-SY5Y cells was inhibited,apoptosis was induced,and immunogenicity was greatly enhanced.

Objective:To construct eukaryotic expressing vector of the full length coding sequence of Bad gene and to express the gene in the basal cell carcinima A431 cells.Methods:Bad gene was amplified from Hela cell line by RT-PCR and the fragment of the cDNA was cloned into eukaryotic expressing vector pcDNA3.1-myc by ligating the fragment into XhoI and EcoRI site.The recombinant plasmid pcDNA3.1-myc-Bad was identified by DNA sequencing and restriction enzyme analysis.The gene transfection mediated by lipofectin was used to introduce the eukaryotic expressing vector of pcDNA3.1-myc-Bad into human basal cell carcinima A431 cells. After selection with G418, resistant colonies were obtained.Trasfection efficiency was identified by Western blot and SABC-FITC assay.Cell proliferation was examined by cell counting and colonogenic assay after transfection.Results:A 500 bp DNA fragment was amplified with RT-PCR.Sequence and restriction enzyme analysis showed that the recombinant plasmid pcDNA3.1-myc-Bad was constructed successfully.In human basal cell carcinima cell line A431 Bad gene was expressed.The cell proliferation was inhibited by 62.6% and colonogenesis by 39.9% by the transfection of the gene.Conclusion: Human Bad gene was successfully cloned.Transfection of basal cell carcinoma cells with the gene may inhibit the cell proliferation and colonogenesis.

Objective To investigate the effects of sodium butyrate (SB) on fetal gastric epithelial cells (GECs) in vitro damaged by N-methyl-N’-nitro-N-nitrosoguanidine (MNNG). Methods The fetal gastric epithelial cells were isolated, cultured and identified. Then the cells of the second generation were cocultured with 10 -5 mol/L, 10 -6 mol/L, 10 -7 mol/L SB for 4 h, then added with 10 -5 mol/L MNNG culturing for another 24 h. After the treated cells were finally cultured in normal culture medium for 40 d, the unscheduled DNA synthesis (UDS), lipid peroxidation (LPO) and ras p21 were detected. The cells were only cultured with 10 -5 mol/L MNNG served as negative control and without MNNG or SB as normal control. Results The level of UDS, the contents of lipid peroxidation products and the concentrations of ras p21 in GECs treated with 10 -5 mol/L and 10 -6 mol/L SB decreased obviously, and had significant differences with that of GECs treated without SB. Conclusion SB can effectively prevent the damage of GECs induced from MNNG.

Objective To explore effective methods of diagnosis and treatment for delayed rupture of the spleen (DRS) through retrospective analysis of 32 cases. Methods A retrospective study was done in 32 cases with DRS. The effects of CT, ultrasound and diagnostic peritoneal lavage on diagnosis of DRS was observed and compared. Results Splenectomy was performed in 26 cases, among which one died four days after splenectomy because of complicated brain injury and six given conservative treatment under continuous monitoring discharged. The mean hospitalization duration was 18.5 days. Conclusions Diagnostic peritoneal lavage can improve the positive rate of peritoneal puncture. Ultrasound is an important method for diagnosing DRS and closely monitoring the patients receiving conservative treatment.

Adenocarcinoma ; genetics ; metabolism ; Adolescent ; Adult ; Aged ; Biomarkers, Tumor ; Child ; Child, Preschool ; DNA-Binding Proteins ; biosynthesis ; genetics ; Female ; Gastric Mucosa ; metabolism ; Genes, Tumor Suppressor ; Humans ; Male ; Middle Aged ; Nuclear Proteins ; biosynthesis ; genetics ; PTEN Phosphohydrolase ; biosynthesis ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Stomach Diseases ; genetics ; metabolism ; Stomach Neoplasms ; genetics ; metabolism ; Tumor Protein p73 ; Tumor Suppressor Proteins

Objective To construct series of reporter plasmids with truncated and deleted hTERT promoter.Methods Gene fragments of hTERT promoter was amplified by PCR and cloned into pGL3-Basic to construct luciferase reporter vectors.Dual luciferase assays were performed with cell lysates of HepG2 and COS-7 cells cotransfected with hTERT promoter reporter plasmids and pRL-TK.Results Series of luciferase reporter plasmids with truncated and deleted hTERT promoter were successfully constructed and respectively named pGL3B-895,pGL3B-371,pGL3B-DELS2,pGL3B-349,pGL3B-329,pGL3B-318,pGL3B-306.Dual luciferase reporter assays showed that all the reporter vectors have promoter activity both in HepG2 and COS-7.Conclusion Series of luciferase reporter plasmids with truncated and deleted hTERT promoter were successfully constructed,and their promoter activity were verified.These plasmids provide necessary experimental naterials for further investigation of regulation of hTERT during hepatocarcinoma development.

Objective To explore a new peptide-based approach independent of HLA to generate antigen-specific CD+ CD8+T cells. Methods Peripheral blood mononuclear cells(PBMC) were stimula- ted for 6 h with IE-1 peptide pool. Then the activated IFN-γsecreting ceils were tested by immunomagnetic selection. And the selected cells were cultured with radio-inactivated PBMC in medium with 100 IU/ml IL-2 for 4 weeks. Results The generated T cell lines consisted of IE-1 specific CD4+ T (6.88%) and CD8+ T cells 92.99%, which demonstrated antigen-specific killing and cytokine secretion. Conclusion T ceils can be proliferated with this new procedure, and maintain its phenotype and antigen-specific function.

Objective To explore the influence factors of cirrhosis complicated with upper gastrointestinal bleeding,and to guide the clinical treatment of patients with cirrhosis and prevent upper gastrointestinal bleeding.Methods One hundred and seventy-five cases patients with cirrhosis and upper gastrointestinal bleeding were treated in the Infectious Disease Hospital of Tangshan and the Affiliated Hospital of North China University of Science and Technology from July 2013 to July 2015 as the case group.One hundred and eighty-two patients with cirrhosis and no upper gastrointestinal bleeding at the same period in hospital as the control group.A face to face questionnaire was used to fill in the questionnaire.Results Multifactor conditional Logistic regression analysis showed that onset season (OR =4.185,95％ CI:1.874-8.354),non steroidal drugs (OR =6.215,95％ CI:2.681-15.532),drinking (OR =5.481,95％ CI:3.205-11.225),portal vein highpressure gastropathy(OR =7.658,95％ CI:3.227-14.714),diameter of portal vein (OR =8.901,95％ CI:1.218-9.026),liver function classification (OR =13.124,95 ％ CI:2.107-15.228) and esophageal varices (OR =11.021,95％ CI:2.181-13.487) were related with patients with liver cirrhosis complicated with upper digestive tract hemorrhage.Conclusion The onset season,nonsteroidal anti-inflammatory drugs,drinking,portal hypertensive gastropathy,portal vein diameter,liver function classification and esophageal varices are the risk fators of liver cirrhosis complicated with upper digestive tract hemorrhage factors.

Aim: To study pharmacokinetics of aconitine,mesaconitine and hypaconitine in rats after single oral administration of decoctions composed with Radix Aconiti Lateralis.Methods: Four groups of rats were orally administered four decoctions including decoction a(Sini decoction),decoction b(decoction composed with Radix Aconiti Lateralis),decoction c(decoction composed with Radix Aconiti Laterali and Radix et Rhizoma Glycyrrhizae Praeparata Cum Melle)and decoction d(decoction composed with Radix Aconiti Laterali and Rhizoma Zingiberis),respectively.Quantitative analysis of aconitine,mesaconitine and hypaconitine in rat plasma was achieved using a liquid chromatography-electrospray ionization/tandem mass spectrometry method.Pharmacokinetic parameters were estimated using DAS 2.0.Results: Pharmacokinetic parameters of aconitine,mesaconitine and hypaconitine were different after oral administration of four decoctions according to Radix Aconiti Laterali combined with different herbal medicines.Multiple peaks were observed in plasma concentration-time curve after oral administration of the decoction of herb couple Radix Aconiti Laterali and Radix et Rhizoma Glycyrrhizae Praeparata Cum Melle,and the results showed a delay in tmax and a prolonger in MRT0-t compared with the decoction of Radix Aconiti Latera-lis.When Radix Aconiti Lateralis was combined with Radix et Rhizoma Glycyrrhizae Praeparata Cum Melle and Rhizoma Zingiberis at the same time in Sini decoction,tmax was delayed too but MRT0-t,was shorten than that of the group of Radix Aconiti Lateralis.Conclusion: The pharmacokinetic parameters of the three compounds obtained in this work shows that the pharmacokinetics of aconitine,mesaconitine and hypaconitine were influenced diversely when Radix Aconiti Laterali was combined with different herbal medicines.

Peptide self-assembled multilayers ( SAMs ) was coated onto the quartz surface. The assembly conditions, such as the assembly agent concentration and assembly time, were examined. The SAMs was characterized via UV-vis absorption spectrometry and scanning electron microscope. Our results showed that the optimal concentration and assembly time for the 3-aminopropyl-triethoxysilane aqueous were 1% ( V/V ) and 3 h respectively, and those of gold nanoparticles were 2. 4 × 10-4 mol/L and 12 h, respectively, while those of peptide solution were 1 × 10-4 mol/L and 12 h, respectively. A new sensitive method, based on the theory that peptide could be cleaved at the site of Arg-Gly by thrombin, was established to detect thrombin. In addition, a good linear relationship was obtained in the range from 2. 8×10-12 mol/L to 9. 9×10-10 mol/L, and the detection limit was 1. 4×10-12 mol/L. The peptide self-assembled multilayers were also used in the analysis of blood serum samples, and the recovery rate was within the range from 91. 6% to 107. 6%.