Objective:To investigate the neuroendocrine properties of adrenalcortical tumors.Methods: We retrospectively reviewed the clinical data of 99 adrenalcortical tumor patients,who were treated in Changzheng Hospital form June 1999 to June 2005.Expression of neuron specific enolase(NSE),chromogranin A(CgA) and synaptophysin(Syn) proteins were examined by immunohistochemistry(S-P method) using monoclonal antibodies.The general data of patients,including the age,symptoms,laboratory findings,and pathological types,were collected and subjected to statistical analysis with SAS v6.12 software.Results: The expression of all the above 3 proteins was found in adrenalcortical adenoma tissues,with the positive rate of NSE being 80%,the positive rate of CgA being 48.9%,and the positive rate of Syn being 75.6%;the positive rates in the adrenalcortical carcinoma tissues were 77.8%,22.2%,and 77.8%,respectively;and those in the normal adrenal tissues was 20%,0%,and 10%,respectively.The positive rates of 3 proteins in adrenalcortical tumors was significantly higher than those in the normal adrenal tissues(P

Objective:To investigate the role of alloreactive T cell in clonal deletion and regulatory T cells ( Treg) in transplant tolerance.Methods:F1 mice were bred by crossing female BALB/c mice and male C57BL/6 mice.Within 24 h,newborn C57BL/6 mice were inoculated with F1 spleen cells via the orbital branch of the anterior facial vein.Six weeks later,the mice were subjected to F1 skin grafting to evaluate their tolerance.Proliferation,flow cytometry and adoptive transfer assay were used to analyze clonal deletion of alloreactive T cells and the expression of CD4+Foxp3+T cells in neonatal treated mice.Results: Newborn C57BL/6 mice injected with F1 splenic cells could induce transplantation tolerance,the level of tolerance was associated with the dose of splenic cells.3×107 splenic cells from F1 mice could induce long-term skin graft acceptance in C57BL/6 mice ,1×107splenic cells significantly prolonged the survival of F1 skin grafts,but the grafts completely rejected within 50 days.The mixed lymphocyte reaction ( MLR) experiment in vivo showed that alloreactive T cell in long-term tolerant mice was deleted completely,but a certain amount of reactive T cells existed in the low-dose group mice.Flow cytometry ( FCM) analysis showed that the expression of CD4+Foxp3+T cell in the high-dose group and low-dose group mice had no obvious difference compared with the naive mice.When alloreactive T-cells were injected into tolerant mice,the skin graft rejection was observed,and Treg cells upregulated in graft-rejected mice.Conclusion:The degree of transplantation tolerance depended on the clonal deletion of alloreactive T cells,instead of on the expression of CD4+Foxp3+Treg cells.CD4+Foxp3+regulatory T cells upregulated in graft rejected mice,which may be served as a negative feedback mechanism to control the intensity of rejection.

Objective To study the effect of phosphodiesterase inhibitor pentoxifylline (PTX) on type 1 diabetes in NOD mice and its possible mechanism.Methods Blood glucose, urinary glucose, insulitis and incidence of diabetes were investigated in NOD mice treated with PTX, insulitis was observed by HE staining and the expressions of IFN ?, TNF ?, IL 10 in pancreas were measured by RT PCR technique. Results The incidence of diabetes in the PTX group was 30.0% which was significantly lower than 67.9% in the control group (P

OBJECTIVE:To optimize kind of safe and effective iodophor thinner.METHODS:Iodophor solutions in several different solvents were made and diluted 5,10 and 20 times,respectively.Then their microbicidal potentials on 3 standard strains were monitored.RESULTS:The standard strains were all killed within 5 min by iodophor solution in normal sodium or by low multiple diluted iodophor solution in distilled water;however,there were a few Escherichia coli and Pseudomonas aeruginosa still alive within 5 min in high multiple diluted iodophor solution in distilled water.Iodophor solution in tap water failed to kill all the standard strains,especially the Pseudomonas aeruginosa.CONCLUSION:Normal saline solution and distilled water have been proved to be the ideal iodophor thinners.

Objective To investigate the expressions of Ki67,p53,CKL,EMA,S-lOO,NSE,CgA,Syn,CEA and nm23 in adrenal tumors and their clinical significance.Methods Clinical data from 157 cases of adre-nal tumor patients were retrospectively reviewed including the clinical informa-tion and pathology data.Expressions of Ki67,p53,CKL,EMA,S-100,NSE,CgA,Syn,CEA and nm23 proteins were studied by immunohistochemistry(SP method)using monoclonal antibodies,and the relationship of their expressions with histopathologic type and clinical imformation was analyzed with SAS v6.12 software.A P value of＜0.05 was considered statistically significant.Results A increase of the expression rate of CKL,S-100,NSE,CgA,Syn and nm23 in adrenal tumors was obsered(P＜0.05).For univariate analysis,the expression of S-100,CgA,Syn was in connection with histopathologie types(P＜0.05).The expression of S-100,CgA,Syn had positive correlation with each other.The expression of CKL,S-100,NSE,CgA,Syn,nm-23 was no difference between adrnalbenign tumors and malignant tumors(P＞0.05),but it was much higher than in normal adrenal tissues(P＜0.05).The expression of Syn in adrenal cortical adenomaa was higher than in adrenal cortical cancers(P＜0.05),the expression of Ki67 in adrenal cortical adenomas was much lower than in adrenal cortical cancers(P＜0.05).The expression of EMA、CKL in adrenal cortical tumors were higher than in adrenal medullary tumors(P＜0.05),the expression of S-100,Syn,NSE,CgA in adrenal cortical tumors were lower than in adrenal medullary tumors(P＜0.05).Conclusions CKL,S-100,NSE,CgA,Syn and nm23 were good markers for adrenal tumors,they could be use for the adrenal tumors diagnosis.Detect Syn and Ki67 simutaneously was helpful to the diagnosis of adrenal cortical tumors.Detect EMA,CKL,S-100,Syn,NSE and CgA simultaneously and combine with clinical data was helpful to diagnosis between adrenal cortical tumors and adrenal medullay tumors.In malignant tumors,blood pressure had positive correlation with the expression of CgA,the size of tumor had neg-ative correlation of blood pressure,no prognostic factor was found.

Objective To investigate the effects of RNA interference (RNAi) targeting angiotensinconverting enzyme (ACE) on blood glucose,insulin resistance,as well as oxidative stress in type 2 diabetic rats.Methods Type 2 diabetic rats were divided into diabetes control group (caudal intravenation with control adenovirus named Ad5),gene treatment group (caudal intravenation with recombinant adenoviral vectors named Ad5-ACE-shRNA,expressing ACE gene-specific shRNA),and enalapril group (intragastric administration with enalapril every day).At the same time,the normal blood glucose control group was set up.All rats were injected two times during the experiment period.Blood glucose was measured before and after the intervention.At the third day of the experiment,expressions of ACE mRNA and protein in pancreas were evaluated by RT-PCR and Western blot,and serum concentrations of ACE and Ang Ⅱ were measured by ELISA.By the end of the experiment,insulin sensitivity index was calculated and expressions of glucose transporter 4 (GLUT4) protein of epididymal adipose tissue and NAD (P) H (p22phox) protein of pancreas were measured.Results Blood glucose levels in the gene treatment group [(17.8 ±1.1) mmol/L] and the enalapril group [(17.9 ± 1.2) mmol/L] were lower than that in the diabetes control group [(24.9 ± 1.3) mmol/L] when the experiment was finished.ACE mRNA and protein expressions in pancreas of the gene treatment group were significantly decreased compared with the diabetes control group (P ＜ 0.05).Serum concentrations of ACE and Ang Ⅱ in the gene treatment group were (16.37 ± 3.01) ng/ml and (18.24 ± 3.69)pg/ml,significantly lower than those of the diabetes control group [(46.67 ± 3.92) ng/ml and (44.93 ± 4.12) pg/ml respectively,both P＜0.05].Insulin sensitivity indexes of the gene treatment group and the enalapril group were (-5.14 ± 0.41) and (-5.17 ± 0.38),being all significantly higher than that of the diabetes control group (-6.18 ±0.46,both P＜0.05).Expressions of GLUT4 protein in epididymal adipose tissue were higher and expressions of p22phox protein in pancreas were lower in the gene treatment group and the enalapril group than those of the diabetes control group (both P＜0.05).Conclusions RNAi targeting ACE gene may delay the progress of hyperglycaemia and improve the situation of insulin resistance and oxidative stress.The RNAi technology may be used as a new strategy of gene therapy for diabetes mellitus.

Objective To investigate the value of mesenteric vascular CTA in the diagnosis of small intestinal neoplasms.Methods A retrospective analysis of mesenteric CTA from January 2008 to April 2013 was conducted in 51 patients with pathologically proven small intestinal neoplasms.Features of intestinal neoplasms CTA signs including neoplasm feeding artery,draining vein,mesangial side vasa recta and the formed neoplasm vessels,were observed.Two radiologists individually used two methods,namely intestinal tumor feeding artery positioning method and Cole fractionation method,for diagnosis and localization diagnosis of tumor,and also for comparing the results with those of surgical pathology.McNemar Chi-square test was adopted to evaluate the diagnosis differences between the two physicians and between the two methods by the same physician.Kappa value was used to assess the consistency of the results.Results Features of intestinal tumors CTA signs:12 cases of enlarged neoplasm feeding artery,9 cases of early displayed draining vein,22 cases of enlarged mesangial side vasa recta,and 11 cases of vessels formed inside and around the neoplasm,single lesion for all and the largest lesion diameter≥ 5 cm for 37 cases.The accuracy of Cole fractionation method positioning and the feeding artery positioning were 84.31％(43/51) and 98.03％(50/51) respectively.Moderate consistency(Kappa=0.49,P＜0.01) was seen with Cole fractionation method by the two physicians and high consistency(Kappa=1.00,P＜0.01) with feeding artery positioning method.McNemar Chi-square test showed no significant difference between the two methods by the same physician and the consistency of the results from the two methods was passable(P were 0.062 and 0.125).Conclusion Mesenteric CTA can display the intestinal tumor feeding arteries and draining veins,and is helpful in identification of the relationship between the tumor and its surrounding blood vessels,which can improve the accuracy of pre-operative localization and qualitative diagnosis for small intestinal tumor.

Objective To investigate the effects of interleukine-22 ( IL-22 ) on the expression of interleukin-6 (IL-6) by rheumatoid arthritis synovial fibroblasts (RASF), and to analyze their association with IL-17+CD4+T (Th17) cells differentiation.Methods RASF were isolated from six patients with rheu-matoid arthritis ( RA) and cultured in vitro.The expression of IL-6 at mRNA and protein levels by RASF were detected by qRT-PCR analysis and ELISA after treatment with different concentrations of IL -22 for dif-ferent periods of time.Anti-IL-22R1 blocking antibody and inhibitor assay were used to analyze the specific receptor and its downstream signaling pathways associated with IL-6 production.IL-22 pre-treated RASF and CD4+T cells were co-cultured for 3 days in the presence or absence of anti-IL-22R1 or anti-IL-6 to measure the percentage of Th 17 cells by flow cytometry .Results The expression of IL-6 by RASF was increased up-on IL-22 stimulation in a dose and time dependent manner (P<0.05), and that was closely related to IL-22R1 and its downstream signaling pathways of p38 and JAK2 (P<0.05).Co-culturing CD4+T cells with RASF and Transwell system indicated that the percentage of Th 17 cells was increased in IL-22 pre-treated group as compared with that in IL-22 untreated group , but it could be down-regulated by either blocking IL-22R1 or IL-6.Conclusion IL-22 promoted the expression of IL-6 by RASF and further enhanced Th 17 dif-ferentiation.Neutralizing IL-22 in synovium of patients with RA might be an effective therapeutic strategy for RA treatment.

Objective To evaluate the application value of inhibitor enhanced modified carbapenemase inactivation method (imCIM) in the detection of class B carbapenemase.The differences between imCIM and EDTA disc potentiation test (EDPT) were comparatively analyzed.Methods A total of 181 strains of carbapenem insensitive strains were collected,among which there were 44 strains of Klebsiella pneumoniae,44 strains of Escherichia coli,43 strains of Acinetobacter baumannii and 50 strains of Pseudomonas aeruginosa.The 83 strains of carbapenem-sensitive strains were composed of 25 strains of Klebsiella pneumoniae,16 strains of Escherichia coli,25 strains of Acinetobacter baumannii and 17 strains of Pseudomonas aeruginosa.The class B carbapenemase in the 264 strains of pathogenic bacteria was screened by imCIM and EDPT,and PCR results were used as gold standard.The statistical analysis wasperformed with consistency check,related-sample Wilcoxon signed rank sum test,independent samples Kruskal-Wallis H test and ROC curve.Results Among the 181 strains of carbapenem insensitive strains,PCR results of 144 strains were positive for drug resistance gene.The samples of class A,B and D of carbapenemase were 39,77 and 28 strains respectively.The results of imCIM showed that 70 strains were positive,and the other 111 strains were negative.The imCIM results of 166 strains were consistent with those of PCR.The results of EDPT showed that 72 strains were positive,and the other 109 strains were negative.The EDPT results of 134 strains were consistent with those of PCR.The results of PCR,EDPT and imCIM of 83 carbapenem sensitive strains were negative.The sensitivity and specificity of imCIM were 85.71％ (66/77) and 97.86％ (183/187),and the value of Kappa was 0.859.The sensitivity and specificity of EDPT were 66.23 ％ (51/77) and 88.77 ％ (166/187),and the value of Kappa was 0.561.The difference of inhibition zone of imCIM (AdimCIM) was different from EDPT(AdEDPr) and the difference was statistically significant (Z =-6.941,P ＜ 0.05).In the imCIM detection,the AdimciM level of class B carbapenemase showed different population distribution position from class A and D carbapenemase with the statistically significant difference (x2 =108.887,P ＜ 0.05).The areas under the ROC curve of imCIM and EDPTwere 0.988 (95％CI:0.977 to0.999) and0.936 (95％CI:0.909 to0.963),respectively.Conclusion imCIM should be accurate,efficient and convenient for screening of carbapenem phenotype for its high sensitivity and specificity,and suitable for epidemiological monitoring.

Objective To Evaluate the performance of serum adenosine deaminase assays of different manufacturers and explore the approach for harmonization of test results.Methods It was evaluated the indice including the limit of blank ,precision,linearity range and reference interval of 10 test systems.It was as the reference system by Mindray test system to evaluate the comparability and the difference of ADA results among 10 different systems.The evaluation was performed before and after calibration by a selected fresh serum assigned by the reference system.A commercial calibrator of the minimum matrix effect was selected from 8 different calibrators as the long-term calibrator to harmonize the ADA results of 10 systems.Results The results of LoB were 0.1-6.3 U/L,respectively.The within-run CVs and total CVs of 10 systems were all less than 5%and actual linearity ranges were conformed to claims of manufacturers.After calibration with fresh serum calibrator ,the averaged difference of 10 test systems was reduced from 14% to 3.0%, and the average difference was 1.8% after calibration with long-term calibrator.The common reference interval of all test systems was 5-24 U/L identically.Conclusions The comparability of ADA measurements can be improved by using a common human serum calibrator and the commutable commercial calibrator.And it is necessary and feasible to develop the standardzation of ADA.