Objective To construct adenovirus vector harboring CTLA4Ig-IRES-CTLA4 gene, and to express the gene in bone marrow mesenchymal stem cells (BMMSCs). Methods The cDNA fragments of CTLA4, and the extracellular domains of CTLA4 and IgGFc were cloned to pT-Easy vectors by RT-PCR from the activated splenocytes of Wistar rats. Recombinant adenovirus vector harboring CTLA4Ig-IRES-CTLA4 was produced from AdeasyTM Adenoviral Vector System by homologous recombination between the Adtrack vector (pAdtrack-GFP-CTLA4Ig-IRES-CTLA4) and the plasmid pAdEasy-1 containing most of the human adenovirus serotype 5 (Ad5) genome in E. coli BJ5183 strain, and then packaged and propagated in 293 cells. BMMSCs were infected with the recombinant adenovirus, and the co-expression of both CTLA4Ig and CTLA4 was then detected by RT-PCR and Western blotting. The immunosuppression function of the transfected BMMSCs was investigated by mixed lymphocyte response (MLR). Results Fusion gene CTLA4Ig was constructed successfully, and the recombinant CTLA4Ig-IRES-CTLA4 adenovirus was then generated by homologous recombination and packaged in 293 cells. The transduction efficiency of recombinant adenovirus in BMMSCs was elevated up to 98.67% determined by flow cytometry using a GFP as marker. CTLA4Ig and CTLA4 were expressed at the same time in BMMSCs detected by RT-PCR and Western blot, and the transfected stem cells showed stronger immunosuppression function than that of BMMSCs or BMMSCs transduced by Ad-CTLA4Ig. Conclusion The BMMSCs transfected by the recombinant adenovirus can co-express CTLA4Ig and CTLA4, which improved the immunosuppression function of BMMSCs in the way of costimulatory pathway blockade.

Background It is well known that sclera remodeling occurs during axial elongation in myopia under the control of growth hormone or its downstream effectors.The role of transforming growth factor-β (TGF-β) in myopia has been determined in previous studies.Bone morphogenetic protein (BMP) is one of members of the TGF-β superfamily,but if it plays an important role in the genesis and development of myopia is not completely clear.Objective This study was to identify the presence of BMPs in normal guinea pigs sclera and investigate the change of BMPs in the sclera in form-deprivation myopia (FDM) of guinea pigs.Methods Thirty young guinea pigs were randomized into normal control group and experimental group using table of random number.FDM models were established by occluding unilateral eyes of guinea pigs with a translucent lens for 14 days in the experimental group,and the fellow eyes served as the controls.Diopter of all eyes was tested by retinoscopy optometry,and ocular axial length was measured by A-sonography before and after modeling.Posterior sclera tissue of the animals was obtained on 14 days,and the relative expression level of BMPs mRNA and protein were assayed by reverse transcription PCR (RT-PCR) and Western blot.The use and care of the animals complied with ARVO Statement.Results On 14 days after occluding of unilateral eyes,the refraction diopter of the experimental group was (-0.48±0.51) D,and that of the fellow eyes was (3.22 ±0.34) D,showing a significant difference between them (t =-12.814,P =0.000).Also,a significant difference in the diopter was seen between the experimental group and normal control group ([-0.48±0.51]D vs.[2.97±0.70]D,t =-11.878,P=0.000).Axial length was (8.30 ± 0.05) mm in the experimental group,(8.11 ±0.06) mm in the fellow eyes and (8.06±0.06) mm in the normal control group,showing a significant increase in the experimental group compared with the fellow eyes and normal control group (t =7.230,P =0.000 ; t =9.084,P=0.000).The expressions of BMP-2 mRNA,BMP-4 mRNA,BMP-5 mRNA in posterior sclera were detected in the normal guinea pigs.Fourteen days after the induction of myopia,the relative levels of BMP-2 mRNA and BMP-5 mRNA in sclera were 0.41 ± 0.11 and 0.65 ± 0.06 in the experimental eyes,which were significantly lower than 0.62 ± 0.07 and 0.84 ± 0.03 in the fellow eyes with the descent range of 34.48％ and 23.67％ respectively (t=2.838,P=0.017; t=2.524,P=0.028).The relative values of BMP-2 protein and BMP-5 protein were 0.44±0.06 and 0.70±0.05 in the experimental eyes,and those of the fellow eyes were 0.61±0.05 and 0.82±0.03,showing significant decline in the experimental eyes with the lowing range of 23.42％ and 15.21％,respectively (t =2.465,P =0.030;t =2.445,P=0.031).No significant differences were found in the expression of BMP-4 mRNA and protein in posterior sclera between the experimental eyes and the normal control eyes (mRNA:t =0.704,P=0.460;protein:t=0.987,P=0.365).Conclusions The expressions of the BMP-2 and BMP-5 in sclera down-regulate significantly in FDM eyes,which suggest that BMP-2 and BMP-5 participate in sclera remodeling during myopia induction.

Diagnosis, Differential ; Humans ; Hyponatremia ; diagnosis ; etiology ; therapy ; Inappropriate ADH Syndrome ; diagnosis ; etiology ; therapy

Objective To investigate the effects of different doses of propofor on ATP-sensitive K~+currents(I_KATP)in human atrial myocytes and the underlying mechsnism.Methods A small piece of myocardiumwas obtained from right atrium in patients undergoing atrial septal defect or ventricular septal defect surgery.Themyocardium specimen was placed in cold Ca~(2+)-free cardioplegic solution aerated with 100% oxygen.Themyocardium specimen was cut into small chunks(1 mm~3).Atrial myocytes were isolated by enzymatic dissociationtechnique.The effects of propofol on I_KATP in atrial myocytes were studied using the whole-cell configuration ofpatch-clamp technique.Results The outward currents were recorded with a pipitte solution containing 0.3mmol?L~(-1) ATP.The currents were inhibited by glibendamide 10 ?mol?L~(-1),a specific K_ATP channel inhibitor,suggesting that the outward currents were I_KATP.I_KATP aws activited by propofol in a dose-dependent manner.Conclusion Propefol can activate the I_KATP in human myocytes in a concentration-dependent manner and themechanism of its myocardial depressant action may be partly explained.

Adult ; Aged ; Female ; Hepatitis B e Antigens ; blood ; Hepatitis B, Chronic ; blood ; drug therapy ; Humans ; Male ; Middle Aged ; Thymidine ; analogs & derivatives ; therapeutic use ; Treatment Outcome ; Young Adult

OBJECTIVETo investigate the correlation between human neutrophil peptide (HNP) and spontaneous bacterial peritonitis (SBP) in order to assess the diagnostic value of quantitative measurement of these alpha-defensins.

METHODSSeventy-seven patients with cirrhosis and ascites were divided into two groups according to diagnosis of SBP (n = 45 with SBP and n = 32 without SBP). Twenty-eight healthy individuals were analyzed as controls. HNP was detected by double-antibody sandwich assay. Peripheral white blood cell (WBC) counts, neutrophil ratio, and levels of procalcitonin (PCT) and C-reactive protein (CRP) were determined by standard methods. Receiver operating characteristic (ROC) curves were used to compare the diagnostic values of HNP, PCT and CRP in SBP.

RESULTSThere were no significant differences between the three groups (SBP, non-SBP, and healthy controls) for WBC count ((6.01+/-2.25)*109 /L, (4.85+/-1.94)*109 /L, and (5.49+/-1.76)*109 /L; F = 2.91, P more than 0.05) and neutrophil ratio (70.70%+/-10.42%, (68.20%+/-8.97%, and 69.50%+/-8.69%; F = 3.07, P more than 0.05). However, compared to the non-SBP group and the healthy controls, the SBP group showed significantly higher levels of HNP ((9.99+/-3.33) ng/ml and (8.92+/-2.30) ng/ml vs. (17.66+/-6.40) ng/ml; q = 3.20 vs. 3.52, P less than 0.05), serum CRP ((15.08+/-9.95) ng/ml and (5.96+/-2.91) ng/ml vs. (31.32+/-18.65) mg/L; q = 11.03 vs. 3.69, P less than 0.05), and positive rate of PCT (25.0% and 10.0% vs. 62.2%; X2 = 10.41 vs. 15.40, P less than 0.0125). The areas under the curve (AUC) showed the following descending trend: HNP more than PCT more than CRP (0.719, 0.707, and 0.629 respectively). Using cut-off points of 10 ng/ml for HNP, 0.5 ng/ml for PCT, and 8 mg/L for CRP, the respective sensitivities for diagnosis of SBP were 71.1%, 62.2%, and 73.3%, the respective specificities were 71.9%, 75.0%, and 56.3%, and the respective Youden's indexes were 0.430, 0.372, and 0.296.

CONCLUSIONHNP is closely related to SBP and can diagnose SBP as reliably as PCT. CRP may help to diagnose SBP, but the results from routine blood testing did not show sufficient statistical significance for diagnosing SBP.

Adult ; Aged ; Bacterial Infections ; blood ; diagnosis ; C-Reactive Protein ; metabolism ; Calcitonin ; blood ; Calcitonin Gene-Related Peptide ; Case-Control Studies ; Female ; Humans ; Male ; Middle Aged ; Peritonitis ; blood ; diagnosis ; microbiology ; Protein Precursors ; blood ; alpha-Defensins ; blood

Objective To explore the clinical application and value of percutaneous transhepatic gallbladder drainage (PTGD)under CT guide in high-risk emergency.Methods In all 57 old patients with high-risk acute cholecystitis in emergency,cuff-PTGD in 39 was performed and fractional step PTGD in 18 was also used.Results PTGD was successfully in all patients.After PTGD,except for 1 patient died of severe cardiac insufficiency,the abdominal pain and fever were alleviated during 72 hours,and the complications was not demonstrated.Conclusion As a safe,noninvasive and accurate method,CT-guided PTGD may relieve symptoms quickly,reduce the mortality and improve the treatment for some old patients with high risk acute cholecystitis.

Lymphocyte function associated antigen-1 (LFA-1) is a member of integrin family, that plays an important role in the adhesion of lymphocytes with other cells and matrix. To investigate the role of LFA-1 in collagen-induced arthritis (CIA), the incidence of CIA, histological and radiological assessments in the LFA-1 deficient (LFA-1~ -/- ) mice and control mice were examined. LFA-1~ -/- mice and control mice were immunized with 100?g collagen type II(CII) emulsified with an equal volume of Freund’s complete adjuvant (CFA), followed by the booster injection of the same amount of CII in CFA on day 21. Then, clinical, histological and radiological assessments were done. It showed that 57% control mice developed arthritis and apparently changed in the histological and radiological assessment, whereas the all of LFA-1~ -/- mice had the normal histological and radiographic response and none developed arthritis. These results suggeste that LFA-1 is indispensable for the onset of CIA.

AIM:To observe the influence of Lycium barbarum polysaccharide (LBP) on the PI3K/Akt/eNOS signaling pathways in ovariectomized rat myocardium .METHODS:Female SD rats (n=30) were divided into sham oper-ation group , ovariectomized group , progynova group , high-dose LBP group and low-dose LBP group .The serum levels of estradiol, lactate dehydrogenase (LDH) and creatine kinase (CK) were measured by ELISA.The myocardial contents of H2 S and oxidative stress injury-related indicators were also detected .The morphological changes of the myocardium were observed with HE staining.The expression of eNOS and PI3K/Akt pathway-related proteins in the myocardium was deter-mined by Western blot .RESULTS: Compared with sham operation group , the serum level of estradiol , the content of H2 S, the activity of GSH-Px, and the expression of eNOS and PI3K/Akt pathway-related proteins in the myocardium in ovariectomized group were all decreased , and the levels of ROS and MDA in the myocardium were increased (P<0.05). The serum levels of LDH and CK were also increased .The arrangement of the myocardial cells was disordered , and the in-tercellular space was also increased in the ovariectomized group .Compared with ovariectomized group , the serum level of estradiol, the myocardial levels of H2S and GSH-Px, and the protein levels of eNOS and phosphorylated Akt were all in-creased in high dose group, while the levels of ROS and MDA in the myocardium were decreased (P<0.05).The serum levels of LDH and CK were also decreased .The morphological changes of the rat myocardium were improved in high dose group.CONCLUSION: LBP prevents and treats postmenopausal cardiovascular lesions through regulating PI 3K/Akt/eNOS signaling pathways in ovariectomized rats .

Objective To compare 18F-FDG、11C-MET and 11C-CHO PET/CT in rat C6 glioma and inflammation models and observe their correlations with HIF-1α and VEGF expressions.Methods Thirtytwo male Wistar rats were included to bear both C6 glioma and turpentine oil-induced acute inflammation (AI).18F-FDG,11C-MET and 11C-CHO PET/CT were performed on rats.The SUVmax ratios of tumor-tomuscle (T/M),AI-to-muscle (AI/M) and tumor selectivity index (SI) were calculated.One-way analysis of variance and two-sample t test were used for statistical analyses.HIF-1α and VEGF expression was detected by immunohistochemical staining.Spearman correlation analysis was performed to evaluate the relationship between T/M ratios and the expressions of HIF-1α or VEGF.Results T/M ratios of 18F-FDG,11 C-MET and 11C-CHO in C6 glioma were 6.89±2.53,2.75±0.87,2.73±1.01,and the AI/M were 4.77±2.21,1.75±0.66,2.23±0.90 respectively.The 18F-FDG and 11C-MET uptake between C6 glioma and AI were significantly different(tFDG =2.133,tMET =3.267,both P＜0.05).The SIMET was significantly higher than SIFDG(t =2.600,P＜0.05).The 11C-CHO uptake between tumor and inflammation showed no significant difference(t=1.537,P＞0.05).T/M ratios of 18F-FDG and 11C-MET were positively related to HIF-1α and VEGF expressions(rs =0.725,0.637,0.621,0.764,all P＜0.05).The T/M ratio of 11C-CHO related only to VEGF (rs =0.682,P＜0.05).Conclusions 18F-FDG and 11 C-MET PET/CT may differentiate C6 glioma from AI,and 11 C-MET PET/CT seems more tumor-selective.11C-CHO PET is less valuable for the differential diagnosis.The 18F-FDG and 11 C-MET uptake of C6 glioma may be related to tumor hypoxia.All the three tracers could reflect tumor angiogenesis,but with different sensitiveness.