The decoy receptor-3 ( DcR3 ) and glutamic acid decarboxylase-65 ( GAD65 ) recombinant adenovirus was construced and transduced into denlritic cells (DC). After the transduced DC were utilized to immunize NOD mice,the CD+8 T cells and blood glucose were analyzed. The results showed that recombinant adenovirus inhibited the proliferation and cytokine release of GAD65 specific T cells,and delayed the incidence of diabetes.Both interferon-γ[ (50.5±7.2)vs(95.4±6.9) and(91.2±6.5) pg/ml] and interleukin-2 [ (46.3±5.1 )vs ( 86.1 ±5.2 ) and ( 80.3 ± 7.3 ) pg/ml ] were decreased compared to those in negative and blank controls ( all P＜0.05 ).The results suggest that DcR3 and GAD65 recombinant adenovirus might provide a promising way for gene therapy of type 1 diabetes.

Objective To investigate the distribution of pathogenic bacteria,drug sensitivity,and the relationship between drug sensitivity and Escherichia coli(E.coli) induced enzymes in childhood diarrhea in the last 2 years in Chongqing area,so as to provide important evidence for pediatric clinical therapy.Methods Thirty-one E.coli induced enzymes,extended spectrum ?-laetamases(ESBLs),cephalosporinase(AmpC)detected in different phenotype methods,and drug sensitivity was measured in paper strip method,and the specimens were collected from children′s hospital affiliated to chongqing university of medical sciences from Jan.2005 to Dec.2006 were determined.Among the total,there were 18 enteropathogeic E.coli(EPEC) strains,8 enterotoxigenic E.coli(ETEC) strains and 5 enteroinvasive E.coli(EIEC) strains.In addition,drug resistance tests by paper strip included chloramphenicol(CHL),amikacin(AMK),gentamicin(GEN),norfloxacin(NOF),ciproflocacin(CIP),cefazolin(CEZ),cefoperazone(CPZ),ceftriaxone(CRO),ceftazidime(CAZ),cefotacime(CTX),cefepime(FEP),imipenem(IPM).SPSS 12.0 software was used to analyze the data.Results Three point two percent of the 31 E.coli were drug resistant to IPM,and 35.5%,38.7% to NOF,CIP individually,but more than 60% to AMK,GEN,even more than 67.7% towards cephalosporin(except ceftazidime and cefepime);the gross enzyme-produced rate was 87.1%,rate of single ESBLs,AmpC,and induction of both enzymes simultaneously presented 64.5%,6.5%,16.1% respectively;and there was marked difference in drug resistance when bacteria that produced single AmpC versus bacteria that produced single ESBLs or that produced both ESBLs and AmpC(Pa﹤0.05).Conclusions The relationships among enzyme′s quantity,sort and bacterial resistance are different.These data show E.coli infected by bacterial diarrhea children in Chongqing due to a high rate of induced enzymes,and their drug resistance vary according to the state of induced enzymes.

New scientific hypotheses detected by mining the potential indirect association inliterature according to the studies on literature-based knowledge discovery are increasinglyapplied in biomedical field and evaluation of literature-based indirect association discovery is a hot spot in recent studies on literature-based knowledge discovery . The role of network characteristics in evaluation of literature-based indirect association discovery during the litera-ture-based knowledge discovery was thus studied.The new indexes for evaluaing the literature-based indirect asso-ciation discovery were esatablished by integrating the co-ocurrent statistic information and the network charateris-tics, which are of greatimportance for improving literature-based knowledge discovery and constructing knowledge discovery system .

Objective To investigate the analgesic effect of salmon calcitonin(sCT)and its effect on expression of calcitonin receptor(CT-R)in periaqueductal gray(PAG). Methods Rat models of neuropathic pain were prepared by chronic constriction injury(CCI). Thermal withdrawal latency(TWL)and mechanical nociceptive threshold(MNT)were measured using hot plate test and yon Frey monofilaments test. The distribution of CT-R in PAG was detected by immunohistochemical method. CT-R protein was quantitatively determined by western blotting. Fourty male SD rats were randomized into 5 groups: normal group, sham-CCI group, CCI group, CCI plussubcutaneous sCT group, and CCI plus microinjection of sCT into PAG group. Results TWL, MNT, andexpression of CT-R in PAG showed no difference between normal group and sham-CCI group(P＞0. 05). TWL and MNT in CCI group were significantly lower than those in normal group(P＜0.05), and expression of CT-R in CCI group was significantly higher than that in normal group(P＜0.05). TWL, MNT and expression of CT-R in CCI rats increased significantly after sCT therapy(P＜0. 05), and the effect was more marked in PAG injection group than subcutaneous injection group(P＜0.05). Conclusions sCT raises the pain threshold and increase the expression of CT-R in PAG of CCI rats, while PAG injection showed more marked effect than subcutaneous injection.

10cm in 11 cases), with an average of 9.2cm. Of all cases,14 were restored by using combined transplantation of mandible itself and a Titanium Support, and another 7 by using only mandible itself. 3 cases have been followed up for more than 1.5 years, 10 cases for 1 -1.5 years, 4 cases for 0.5-1 years, another 4 cases less than 0.5 years. The results of reconstruction, from the stand-point of both shape and function, were satisfactory.

In an attampt to establish the functional expression of STGC3 with doxycycline (Dox) induced Tet-onregulating system in human nasopharyngeal carcinoma cell line CNE2, an ideal experimental platform wasprovided for further studies of STGC3. pTet-on regulating plasmid was transfected into CNE2, and stableexpression of Tet-on was established in CNE2 through G418 select. Then the response plasmid of recombinantpTRE-STGC3 was steadily transfected into positive CNE2/Tet-on cells with hygromycin screen. Dox was used toinduce the expression of STGC3 and a cell clone sensitive to Dox was selected. The best-induced concentrationwas determined with different concentration of Dox induction. Growth curves, clone formation rate and cell cycledistribution were detected after STGC3 gene up-regulated expression with Dox induction. The growth capacity andclone formation potential of CNE2/Tet /pTRE-STGC3 was significantly suppressed, compared with the controls(P

Background Retinal pigment epithelium (RPE) cell transplantation is the primary means of human trial for the treatment of retinal degeneration.Induced pluripotent stem cells (iPSCs) will become an important source for cell transplantation.In addition,iPS-RPE cells may provide a personalized treatment platform for the patient's own cells treatment.Objective This study was to evaluate the feasibility of human fibroblasts differentiate toward iPSCs from retinitis pigmentosa (RP) patients and toward iPSC-RPE cells from non-RP individual by retroviral transfection of Oct4,Sox2,c-Myc and KLF4 genes.Methods Human thigh skin tissues were obtained from a RP patient with hotspot mutation of SNRNP200 p.S1087L and individual without SNRNP200 p.S1087L mutation,respectively,with the size 2 c m×3 cm.Human dermal fibroblasts were isolated and cultured by trypsin digestion and explant method.The fibroblasts were transfected by a series of retrovirus and cultured by human embyonic cellconditioned medium to generate and induce iPSCs,and then the iPSCs were identified by morphology,alkaline phosphatase (AP) staining and immunofluorescence assay of specific markers of pluripotent stem cells.iPSCs suspension were injected into SCID mouse to observe the tumorigenesis.The iPSCs from non-RP subject were induced to differentiate toward iPS-RPE cells by embryonic body (EB) inducing method,and iPS-RPE cells were identified by detecting the expression of RPE65,zonula occludens protein 1 (ZO-1) and lecithin retinol acyltransferase (LRAT).Results Cultured human dermal fibroblasts showed fusiform or polygon shape and intercellular close arrangement,and Vimentin was positively expressed in the cells.Small cell colonies were harvested 5-7 days after infected by retroviruses,and the morphology changed from spindle into round mass.The hESC-like iPSCs clonies appeared 20 days after cultivation,and the positive expressions of hESC-specific surface antigens including SSEA3,SSEA4,TRA-1-60,TRA-1-81 and Nanog were found in the cells 25-30 days after cultivation,and the positive staining of AP was obtained 12 weeks after cultivation.A teratoma was formed at the injection site of iPSCs suspension in SCID mouse.Immunofluorescence technique showed that RPE cell-specific proteins including RPE65,ZO-1 and LRAT proteins were positively expressed in iPS-RPE cells at 30 days after differentiation.Conclusions Mutation SNRNP200 p.S1087L of RP patient-specific iPSCs can be induced from human dermal fibroblast by retrovirus infection method.The function and morphology of the iPSCs were similar to hESCs.Human iPSCs cell line generated from the dermal fibroblasts of non-RP individuals can differentiate into iPS-RPE cells.

Objective To investigate the neuroprotective effect of thyroid hormones T3 on cerebral ischemia-reperfusion injury in rats and its mechanism.Methods SD rats were divided into four groups:sham+saline group,sham+T3 group,MCAO+saline group,MCAO+T3 group.The cerebral ischemia-reperfusion injury rat models were established by right middle cerebral artery occlusion.Thyroid hormones(10 μg/100 g)or normal saline were given respectively by intraperitoneal injection twice at 1 h after the onset of ischemia and 6 h after reperfusion.Neurobehavioral score was evaluated at 24 h after reperfusion;TTC staining was used to label infarction area;RT-PCR was used to detect the mRNA level of nerve growth factor(NGF)and brain derived neurotrophic factor(BDNF)in brain tissue;Western blot was employed to determine alterations in protein levels of NGF and BDNF.Results Compared with MCAO+saline group,the neurological deficit and the volume of cerebral infarction of MCAO+T3 group was decreased,and the mRNA and protein expression of NGF and BDNF of MCAO+T3 group were increased(P<0.05).Conclusion Thyroid Hormones could promote the nerve repair,stimulate the nerve regeneration and improve the nervous behavioral function by up-regulating the expression of NGF and BDNF.

Objective To investigate a new method which could not only avoid the extrusion of the silicon implant, but also be benefit for the reconstruction of nasal tip and alar during nasal augmen-tation procedure. Methods Folded lower lateral cartilage flap combined with silicon implant was ap-plied for nasal dorsal augmentation and reconstruction of nasal tip and alar with its unique character at the same time. Results 12 cases were all primary healing, without infection, extrusion of implant and other complications. One month after operation, the projection of nasal tip was increased, which had better delicate and definite shape, natural appearing tip adding contour and height to nasal tip, pleas-ant definition. Conclusions Folded lower lateral cartilage flap combined with silicon implant is an ef-fective method to decrease the incidence of extrusion, when we want to have a little over-projected na-sal tip with prosthesis. It is also helpful for reconstructing the nasal tip and alar with its unique char-acter.

OBJECTIVEGas chromatography (GC) was used to investigate the cellular fatty acid (CFA) composition of 141 Acinetobacter baumannii and 32 A. calcoaceticus isolates from different locations in China and to find chemical markers to differentiate these two closely related bacteria.

METHODSWhole cell fatty acid methyl esters (FAMEs) were obtained by saponification, methylation, and extraction for GC analysis, followed by a standardized Microbial Identification System (MIS) analysis.

RESULTSAll A. baumannii and A. calcoaceticus strains contained some major fatty acids, namely, 18:1 ω9c, 16:0, Sum In Feature 3, 12:0, 17:1ω8c, 3-OH-12:0, 17:0, Sum In Feature 2, 2-OH-12:0, and 18:0 compounds. Although most of the total CFAs are similar between A. baumannii and A. calcoaceticus strains, the ratios of two pairs of CFAs, i.e., Sum In Feature 3/18:1 ω9c versus 16:0/18:1 ω9c and Sum In Feature 3/18:1 ω9c versus unknown 12.484/18:1 ω9c fatty acids, could differentiate these two closely related bacteria. A. baumannii could be easily classified into two subgroups by plotting some ratios such as Sum In Feature 3/16:0 versus 17:0 and Sum In Feature 3/2-OH-12:0 versus 17:0 fatty acids.

CONCLUSIONThe ratios of some CFAs could be used as chemical markers to distinguish A. baumannii from A. calcoaceticus.

Acinetobacter baumannii ; classification ; cytology ; metabolism ; Acinetobacter calcoaceticus ; classification ; cytology ; metabolism ; Biomarkers ; metabolism ; Fatty Acids ; metabolism ; Species Specificity