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 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 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 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 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 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 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 observe the clinical therapeutic effect of hyperbaric oxygen (HBO) therapy and swallowing training combined with Fengchi acupoint acupuncture on dysphagia caused by pseudobulbar paralysis after stroke. Methods Seventy-five patients were randomly divided into control group, HBO therapy group, swallow training group, acupuncture group and combined treatment group (n = 15). Patients in all groups were given cerebro-vascular disease routine treatment. In addition, the patients in HBO therapy group, swallowing training group and ac-upuncture group were given HBO therapy, swallowing training and Fengehi (GB20) acupoint acupuncture simultane-ously, respectively. The patients in combined treatment group were given HBO therapy combined with swallowing training and Fengchi acupoint acupuncture. Modified Kubota-Chiari's swallowing functional classification was used to assess swallowing function before and on the 3rd d,6th d,9th d, 12th d, 15th d, 18th d and 21st d after treatment. Results It was shown that the patients in all groups but the control group got significant improvement in swallowing after treatment, as demonstrated by the swallowing function scores (P < 0.05). Moreover, the priority order in im-provement of swallowing function and recovering rate were: combined treatment group > acupuncture group > swal-low training group > HBO therapy group > control group(P < 0.05). Conclusion The combination of HBO ther-apy, swallowing training and Fengchi acupoint acupuncture could act synergically to ameliorate dysphagia caused by pseudobulbar paralysis after stroke.

Objective To investigate the feasibility of modified rapid Carba NP test for the detection of carbapenemase,and analyze the differences between the modified method and Carba NP test.Methods A total of 264 strains of gram-negative bacillus,including 164 carbapenem-resistant strains and 100 sensitive strains,were collected,and their carbapenemase were detected by Carba NP test and the modified rapid Carba NP test,respectively.The differences between the two tests were evaluated based on PCR as a reference.Results Among 164 carbapenem-resistant strains,carbapenemase gene was detected in 144 strains by PCR.The carbapenemase gene was negative in 100 sensitive strains.Among 164 carbapenem-resistant strains,135 were positive for the Carba NP test,while 130 for the modified rapid Carba NP test.One hundred of sensitive strains were negative for the two Carba NP tests.Compared with the results of PCR,the sensitivity,specificity and Kappa value of the Carba NP test were 91.7％ (132/144),97.5％ (117/120) and 0.886,respectively,while those of the modified rapid Carba NP test were 89.6％ (129/144),99.2％ (119/120) and 0.879,respectively.There was no significant difference in the positive rates between Carba NP test and the modified rapid Carba NP test (x2 =1.45,P ＞ 0.05).Conclusion The modified rapid Carba NP test which has high consistency with the PCR method,is faster and cheaper than the Carba NP test,and may be applied to epidemiologic survey and the early monitoring of nosocomial infections.