Objective To evaluate the diagnostic potential of capsule endoscopies (CE) for suspected small bowel diseases. Methods From September 2002 to June 2003, 23 patients (12 males, 11 females ) , age ranged 10 to 75 years old, with suspected small bowel diseases, were referred to our department to perform CE using Given M2A Video Capsule System. All patients had undergone previous negative endo-scopic evaluation at least once with esophago-gastro-duodenoscopy and total colonoscopy. Additional diagnostic work-up including small bowel enteroclysis, selective angiography, scintigraphy and puch enteroscopy, were performed, totally 36 procedures. Two endoscopists independently reviewed capsule images to arrive a consensus diagnosis. The initial diagnostic yield was quantified, and the value of CE was assessed. Of the 23 patients, 18 suffered from obscure gastrointestinal bleeding, 3 abdominal pain, and 2 chronic diarrhea. Results Twenty-four studies in 23 patients were evaluated. During CE patients have not complained of any uncomfortable feeling, only one patient had repeated the procedure because the capsule lodged at the inferior segment of esophagus near the Z line. Of the 23 patients, 20 had positive findings with a diagnostic yield of 86. 8% . The positive findings included inflammatory lesions in 10 patients (Crohns disease 7 , aphthous ulcer 3 ) , vascular abnormalities 9 ( phlebectasis 6, angioma 2, angiodysplasia 1) , submucosal nodulation 2 , diverticula 2, jejunal stromal tumor 1. Four of 23 patients had more than one lesion. Diagnosis of 6 patients was confirmed by surgery and /or pathology. Nineteen capsules passed in the direction with the camera facing forward, while the other 4 backward. Generally, it delayed in passing through the pylorus and ileocecal valves. Capsules reached cecum in 17 patients (73. 9% ) . Conclusion CE provided clearly the small intestinal images, and is an efficient tool in diagnosing small bowel diseases with a high diagnostic yield.

Objective To investigate the effects of oral recombinant attenuated Salmonella ophimurium urease B subunit and catalase in the treatmeat of H.pylori infection in a H.pylori infected mouse model. Methods Thirty C57BL/6 mice were randomized into three groups and challenged twice by oral administration of H.pylori in three days. Four weeks after the second challenge, the mice were immunized by oral administration of recombinant attenuated Salmonella typhimurium urease B subunit (group A), recombinant attenuated Salmonella typhimurium catalase (group B), or saline (group C) respectively, and all mice were sacrificed 4 weeks after the immunization. The stomachs were collected for rapid urease test, modified Giemsa stain and quantitative culture to observe the gastric H.pylori densities, and HE stain was performed to assess the presence of inflammation. Lymph cells from the spleens were served for lymphoproliferation assay. Results Gastric H.pylori densities of group A, B and C were 1.58?10 5 CFU/g, 4.88?10 5 CFU/g and 1.92?10 6CFU/g respectively. H.pylori densities of therapeutic groups were significantly decreased ( P

Objective Compared with corresponding monovalent vaccine and pure vaccine vector, live attenuated Salmonella typhimurium expressing bivalent antigens of Helicobacter pylori (H. pylori) , UreB/HpaA was constructed by introducing a flexible and tenacious linker into urease subunit B (UreB) and HpaA of H. pylori and its protection against H. pylori infection in mice was then investigated. Methods UreB/hpaA fusion gene was amplified form H. pylori genomic DNA using sequence overlap extension PCR (SOE-PCR). UreB/HpaA bivalent live vaccine was constructed using attenuated Salmonella typhimurium vaccine vector SL3261, and its stability in vivo was observed in C57BL/6 mice. Evaluation of protection against H. pylori infection was performed in native female C57BL/6 mice by oral immunization with a single dose of the live SL3261 vaccine strain expressing UreB/HpaA (10 8 CFU). Results Sequencing results showed that encoding sequence (GGTGGAGGC) of three glycine residues was inserted into the position between ureB and hpaA fusion gene as an adapter. Bivalent live vaccine strain could be recovered from spleen and Peyer's patches for a longer time (at least 10 days). Bivalent live vaccine induced marked elevation of the levels of serum specific IgG1 and IgG2A in mouse. The immune protection rate of UreB and HpaA bivalent live vaccine was 77.3% (17/22). However only 50.0% (12/24) of the mice immunized with SL3261 vaccine strain expressing UreB and 43.5% (10/23) of the mice immunized with SL3261 vaccine strain expressing HpaA were completely protected against H. pylori challenge. Conclusions Oral immunization of mice with bivalent UreB/HpaA live vaccine could induce protective immunity against H. pylori , and the protection rate of bivalent vaccine appears to be higher than that of monovalent vaccine.

Objective:Investigated the effect of recombinant urease A subunit and catalase vaccines on protection against Helicobacter pylori (HP) infection Methods:Recombinant plasmids expressing UreA or KatA were constructed Expression was induced with IPTG,analyzed by SDS PAGE and Western blot UreA and KatA were purified with Bulk GST purification module Results:The recombinant plasmids could express GST UreA and GST KatA fusion proteins,which accounted for 35% and 19% of total bacteria proteins respectively,and could specially react with antibody against GST The purity of the purified UreA and KatA was over 95% Conclusion:The work laid a foundation for further studies of HP vaccine The recombinant vaccines may play an important role in preventing and treating HP infection and related diseases

OBJECTIVETo construct and identify a lentiviral vector of RNA interference targeting human Notch-1 gene.

METHODSTo determine the Notch-1 gene sequences, three RNAi target sequences (shRNA1-3) were designed in accordance with the RNAi sequence design principles and cloned into the lentiviral vector pLenOR-THM by endonuclease BamH I restriction, EcoR I double digestion, and T4 DNA-ligase ligation. After the transformation into competent DH5alpha bacteria, the candidate clones were identified by Kpn I and EcoR I double digestion and DNA sequencing. The recombinant and three packaging plasmids were co-transfected into human embryonic kidney cell line 293T cells by lipofectamine to produce the lentiviral particles. The viral titer was determined. The 293T cells were infected by the lentiviral particles obtained, and transfection efficiency was assessed using a fluorescent microscope. The lentiviral vector particles were also transfected into ACC-M cells. The Notch-1 expression in the transfected cells was assayed by quantitative reverse transcription polymerase chain reaction (QRT-PCR) and Western blot analysis.

RESULTSThe lentiviral RNAi vector pLenOR-THM-Notchl for Notch-1 gene was constructed successfully. Strong green fluorescence was observed in the 293T cells under fluorescent microscope after co-transfection of the cells with the four plasmids of lentiviral vector. The virus in the supernatant reached a titer of 5.8 x 10(8) TU x mL(-1). The transfection efficiency of the collected virus exceeded 90% in 293T cells with 1 as a multiplicity of infection. The third lentiviral vector was found to significantly inhibit the Notch-1 expression at the mRNA and protein levels.

CONCLUSIONThe lentiviral RNAi vector of Notch-1 has been successfully constructed and identified.

Cell Line ; Genetic Vectors ; Humans ; Lentivirus ; Plasmids ; RNA Interference ; RNA, Messenger ; RNA, Small Interfering ; Receptor, Notch1 ; Transfection

AIM: To study the role of endothelin-1 (ET-1) in portal hypertension (PHT) induced by endotoxin. METHODS: Collagenase in situ perfusion was adopted to separate hepatic stellate cells (HSCs). HSCs was cultured on concretized collagen. ET-1 anti-sense oligonucleotide was added into the culture medium and then LPS was also added up to the concentration of 1 000 ?g/L. The diameters of the concretized collagen were measured. Sense and mis-sense oligonucleotide were applied as control. ET-1 in the culture medium was detected by radioimmunoassay and ET-1 mRNA in HSCs was detected by RT-PCR. ?-actin of HSCs was detected by Western blotting. RESULTS: The diameter of concretized collagen on which HSCs pretreated with ET-1 anti-sense oligonucleotide was 93.3%?3.8% the size of the primary. The diameter of concretized collagen of the control groups were 70.1%?4.8% and 70.5%?3.9% (P

Objective To construct a recombinant live attenuated Salmonella typhimurium vaccine strain expressing Helicobacter pylori urease subunit B (ureB).Methods ureB gene was amplified by PCR and cloned into a prokaryotic expression plasmid pTrc99a, and the identified recombinant plasmid was then used to transform an attenuated Salmonella typhimurium vaccine strain SL3261. The ureB expressed in the recombinant vaccine strain was analyzed by SDS-PAGE and optical density scanning. Two and 10 days after recombinant strain intragastric immunization, the C57BL/6 mice were sacrificed, and the spleens and terminal ileums were cultured.Results The ureB gene could be amplified from the recombinant prokaryotic expression plasmid pTrc99A-ureB and the plasmids extracted from transformed SL3261 strain. SDS-PAGE and optical density scanning indicated that ureB was expressed in the recombinant vaccine strain SL3261 (pTrc99A-ureB) as a protein with 66*!kD of molecular weight. Recombinant strain was found in both spleen an terminal ileum of each mouse two and ten days after intragastric immunization.Conclusions A recombinant liver attenuated Salmonella typhimurium vaccine strain expressing Helicobacter pylori ureB was constructed and identified, and this study will help to develop an oral recombinant live vaccine against Helicobacter pylori infection.

Abstract Physical health of contemporary children and adolescents decreasing due to physical inactivity. After review of the implementation of physical activity promotion among children and adolescents at home, this paper analyzes the possible reasons of physical activities neglected, constrained and occupied by the family, school and community, and proposes an integrated supportive environment for physical activities among "family school community", so as to promote physical activity among children and adolescents and improve their physical health accordingly.

Objective To investigate the efficacy of implantation of radioactive iodine-125 (125I) seeds combined with chemotherapy in treatment of stage Ⅲb and Ⅳ non-small cell lung cancer (NSCLC).Methods Ninety patients with stage Ⅲb and Ⅳ NSCLC were divided into two groups.The combined group (n=43) received chemotherapy with TP (paclitaxe plus cispla tin) or GP (gemcitabine plus cisplatin) 3 days after 125I particles implantation,and the control group (n=47) only received TP or GP chemotherapy.The effective rate,1-year and 2-year survival rate,the median survival time were compared between the 2 groups.Results The total effective rates of the combined group and the control group of stage Ⅲ b NSCLC were 84.00％ and 48.28％,and the effective rates of stage Ⅵ NSCLC were 72.22％ and 33.33％ (both P＜0.05).The 1-year and 2-year survival rate of the combined group and control group of stage Ⅲb NSCLC were 67.80％,36.00％ and 37.90％,13.83％,respectively (both P＜0.05).The median survival time was 15.7 months and 8.6 months.However,the survival rates of the combined group and the control group of stage Ⅳ NSCLC were 44.44％,16.70％ and 22.22％,11.10％,respectively,while the difference was not statistically significant (P＞0.05).The median survival time was 8.9 and 6.0 months.Conclusion The implantation of radioactive 125I seeds combined with chemotherapy can obtain a significant efficacy in the treatment of stage Ⅲb and stage Ⅳ NSCLC.For patients with stage Ⅲb NSCLC,combined therapy can improve the survival rate.

Objective:To evaluate the influence of different detector widths and signal acquisition positions of wide-detector CT in different scanning modes on CT number and noise, and to provide a basis for reasonable selection of scanning modes and related parameters in clinical practice.Methods:The body dose phantom was scanned by GE Revolution CT. The scan was performed with detector widths of 40, 80 and 160 mm in sequential scanning mode and with detector width/pitch combinations of 40 mm/0.516, 40 mm/0.984, 80 mm/0.508 and 80 mm/0.992 in spiral scanning mode. The phantom was placed at the central and peripheral of the selected detector widths, and the adjacent positions between two axial scans. The images of the phantom were evaluated subjectively and the CT numbers and SDs were measured. The differences between the measured values at different imaging parameters were compared. The multi-group Friedman test was used to compare CT numbers and SD under different scanning parameters in sequential scanning mode, and the Wilcoxon test was used to compare CT numbers and SD in spiral scanning mode.Results:There was no statistically significant difference in the geometric shapes of the phantom images obtained at any combination of parameters. In sequential scanning mode, the differences at different detector widths were statistically significant (χ 2=14.00, P=0.001) with CT numbers at 40 mm and 160 mm greater than CT numbers at 80 mm ( P<0.05). The differences at different signal acquisition positions were statistically significant (χ 2=12.04, P=0.002) with CT numbers at peripheral and adjacent greater than CT numbers at central ( P<0.05). In spiral scanning mode CT numbers at detector width at 80 mm were greater than CT numbers at 40 mm ( Z=-2.10, P=0.036). For SD, the differences at different detector widths were statistically significant in sequential scanning modes (χ 2=8.17, P=0.017) with SD at 160 mm greater than SD at 80 mm ( P<0.05). The differences at different signal acquisition positions were statistically significant (χ 2=13.50, P=0.001) with SD at peripheral greater than SD at central ( P<0.05). In spiral scanning mode SDs at pitches 0.984 and 0.992 were greater than SDs at 0.516 and 0.508 ( Z=-2.66, P=0.008). There were no significant differences among other groups. Conclusion:The selection of scanning mode, detector width and signal acquisition position of wide-detector CT will affect the image CT numbers and SDs.