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.

BACKGROUND: Genistein is the main component of phytoestrogen soy isoflavone and its structure is similar to estrogen,which suggests that it might prevent or delay osteoporosis. Research on the effects of genistein on bone mineralization and calcium(Ca), phosphor(P), zinc(Zn) and magnesium (Mg) in ovariectomized rats are rare.OBJECTIVE: To investigate the effects of genistein on bone mineralization and Ca,P,Zn and Mg in ovariectomized rats to provide a theoretical gist for the prevention of osteoporosis by genistein.DESIGN: A controlled experiment based on experimental animals.SETTING: Department of Nutrition,General Hospital of Chinese PLA.MATERIALS: The study was conducted in the Institute of Hygiene and Environmental Medicine,Academy of Military Medical Sciences of PLA between February and June 2003. Ten-week old female Wistar rats [certification number: (military medical animal): D98014] with a body mass of(170±20) g were selected.INTERVENTIONS: Experimental animals were fed with normal feeding for 6 weeks and then the feeding was changed to AIN-93 compound. Animals were then randomly divided into ovariectomized group (n=40) and sham-operation group(n=7) based on bodyweight after 5 days. Ovariectomized group received ovariectomy and sham-operation group only received abdominal incision. After 5 days of recovery,the ovariectomized group was further randomly divided into 5 subgroups with 8 rats each including ovariectomized control subgroup,estrogen subgroup [diethylstilbestrol 20 μg/(kg · d)],genistein Ⅰ,Ⅱ,or Ⅲ subgroup[dose of 25,50 or 100 mg/(kg · d)]. After 3months of feeding,6 rats were randomly selected from each group for the detection of bone density and corresponding bone hismorphometric indicators.MAIN OUTCOME MEASURES: Bone density,corresponding parameters of bone mineralization,Ca,P,Zn and Mg contents in bone of mice in each group RESULTS: After ovariectomy,femoral bone density decreased [(0. 247± 0.007) g/cm2],mean osteoid width increased[(7. 04 ±0. 32)μm],bone mineralization delayed [(4.96±0.99) days],osteoid maturity prolonged [(26.99±7.70) days],and Ca[ (251.11± 5.31) mg/g],P[(115.08± 3.78) mg/g],Zn[ (299.69±37.1)μg/g] and Mg[(4. 32±0. 12) μg/g]were all significantly different from that of sham-operation group(P<0.05).After the application of genistein,femoral bone density had a tendency of improvement[ (0. 250±0. 007) g/cm2],mean osteoid width narrowed[ (4. 97±0.77) μm],bone mineralization delayed time[ (3.18±0.69) days] and osteoid maturity time[(14.53 ±3.84) days] shortened, contents of Ca [(270.00±5.65) mg/g],P[(124.25±2.37) mg/g] andMg[(4.61±0. 08) μg/g]elevated while Zn content had no significant changes.CONCLUSION: Genistein promotes osteoid mineralization,reduces the loss of Ca, P and Mg in the bone and prevent the generation of osteoporosis in unsexed rats

Objective To investigate the clinical effects of biofeedback therapy on constipation-predominant irritable bowel syndrome(IBS-C).Methods 30 patients with IBS-C were given to biofeedback therapy,5 times every treatment course.and then their clinical symptoms,mental state and quality of life before the treatment and at the end of treatment with biofeedback therapy were respectively evaluated by symptom scores,self-rating anxiety scale (SAS),self-rating depression scale (SDS),and short form 36 health survey questionnaire (SF-36).Results Post-treatment with biofeedback therapy,there were very significant decrease in total and subscales scores of bowel symptom including abdominal pain/discomfort,abdominal distension,abnormal appearance of defecation,abnormal process of defecation((12.31±2.01) vs (19.16±2.17),(2.95±0.57) vs (5.04±1.04),(2.64±0.92) vs (4.25±1.09),(3.66±1.09) vs (5.10±0.57),(3.06±1.08) vs (4.77±0.95) respectively,P＜0.01).The integration of SAS and SDS were obviously decreased after treatment((39.53±6.39) vs (44.43±7.89),P＜0.05;(40.70±8.38) vs (46.46±8.74),P＜0.05),the SF-36 scores were also improved in five dimensions including rolephysical,social-functioning,vitality,role-emotional and mental health((74.16±21.25) vs (57.0±39.40),(86.21±13.54) vs (75.54±20.96),(75.16±13.42) vs (64.66±20.54),(78.87±28.36) vs (57.76±46.26),(81.60±16.08) vs (71.20±22.04) respectively,P＜0.05).Conclusion Biofeedback therapy can improve the clinical symptoms,alleviate the moods of anxiety and depression,and improve the quality of life in patients with IBS-C.