Objective To validate and compare the two spinal microdialysis techniques: a linear tissue probe (LM-3) in the spinal dorsal horn and a loop probe in the cerebral spinal fluid (CSF) in determining peripheral nociceptive stimulation-evoked glutamate (Glu) release in the spinal cord of freely moving rats. Methods Twenty-eight adult male Wistar rats weighing 300-350 g were randomly divided into two groups: in group A a LM-3 probe was implanted into the spinal dorsal horn and in group B a loop probe was placed in the CSF. Twenty-four hours after the implantation of the probe, microdialysis was initiated with perfusion of modified Ringer' s solution at a low flow rate of 5 ?l?min-1 . Following an 1 h equilibration phase the baseline Glu concentrations were measured every 10 min for 1 h. Thereafter 50 ?lof 5% formalin was injected into one hindpaw of the rats and samples were collected every 10 min for 90 min. Furthermore 8 rats in group A were further divided into 2 subgroups to investigate the effects of the flow rate of microdialysis and composition of perfusate on the baseline Glu release.Results The baseline levels of Glu were (0.82?0.09) ?mol?L-1 with LM-3 probe and (5.96?0.22) ?mol?L-1 with the loop probe. In group A (LM-3 probe) when the flow rate of the modified Ringer's solution was decreased from 5 to 2 ?l?min-1 the extracellular Glu concentrations were increased to 223%?7% of the baseline (n = 4) , whereas perfusion with artificial CSF reduced Glu concentrations to 62% ?10% of the baseline (n = 4) . Injection of formalin into the hindpaw induced a short-lasting but significant increase in Glu concentration with a similar profile and time course using either of the two microdialysis approaches. Conclusion Microdialysis in the dorsal horn or in the CSF are both effective techniques to assess Glu release in the spinal cord of rats. Peripheral nociceptive input induces a short-lasting increase in Glu release with a similar profile and time course using either of the two microdialysis approaches. The microdialysis of the dorsal horn provides a useful tool to precisely locatewhere the release of the neurotransmitter occurs, whereas the loop probe in CSF is more reproducible for simultaneous investigation of drug effects.

Objective To analyze and evaluate the expression and significance of costimulatory molecules CD 28 ,CTLA-4 ,CD86 , CD80 mRNA in peripheral blood mononuclear cells of the patients with HBV chronic change .Methods The levels of costimulatory molecules CD28 ,CTLA-4 ,CD86 ,CD80 mRNA in peripheral blood mononuclear cells were detected in 24 cases of chronic hepatitis B (CHB) ,24 cases of liver cirrhosis(LC) ,28 cases of hepatocellular cancer(HCC) and 30 normal control(NC) subjects by real time quantitative PCR .Results Compared with the NC group ,costimulatory molecule CD28 mRNA level in the CHB group was signifi-cantly decreased(t= -2 .11 ,P<0 .05);CTLA-4 mRNA level in different diseases groups was decreased to different degrees :the CHB group(t= -2 .52 ,P<0 .05) ,the LC group(t= -2 .11 ,P<0 .05) and the HCC group(t= -2 .56 ,P<0 .05);CD86 mRNA level in different diseases groups was decreased to different degrees too :the CHB group(t= -3 .68 ,P<0 .01) ,the LC group(t= -2 .99 ,P<0 .01) and the HCC group(t= -4 .42 ,P< 0 .01);CD28/CTLA-4 mRNA level was significantly increased in the HCC group(t= 2 .12 ,P< 0 .05);CD80/CD86 mRNA level was significantly increased to different degrees with the progress of HBV chronic change:the CHB group(t=2 .10 ,P<0 .05) ,the LC group(t=2 .59 ,P<0 .05) and the HCC group(t=3 .74 ,P<0 .01) . Conclusion The expression abnormality of CD28/B7 family costimulatory molecules mRNA in HBV infectious patients may be closely related with the immune dysfunction and the development and progression of the chronic change .

Objective To investigate the effects of tumor-necrosis factor α (TNF-α) on the expressions of proprotein convertases (PC) and its relationship with the inhibition of hepatitis B virus (HBV) replication.Methods HepG2.2.15 cells cultured routinely were exposed to 20 μg/L recombinant TNF-α and/or 20 μmol/L PC inhibitor (DEC) for 18 h.Then Followed cells werecollected and cell total RNA and HBV DNA were extracted.PC mRNA and core-associated HBV DNA were measured using real-time polymerase chain reaction (PCR) techniques.Measurement data was compared using t-test.Results When PC mRNA expressions in the blank group was as to 1,the expressions of PC1/3、PC2、furin、PC4 、PC5/6 、PACE4 and PC7/8 mRNA in HepG2.2.15 cells treated with 20μg/L TNF-α treatment for 18 h were all up-regulated,which were 3.3±0.7、79.3±3.3、77.5±1.3、19.2±3.1、1.3±0.1、1.4± 0.2、274.8± 7.1,respectively (all P＜0.05).Treatment of 20 μg/L recombinant TNF-α for 18 h significantly reduced core-associated HBV DNA compared with blank gourp (0.21∶1,t =8.79,P =0.002),while 20 μmol/L DEC significantly up-regulated core-associated HBV DNA (3.84∶ 1,t=7.67,P=0.004).Moreover,core-associated HBV DNA in group of DEC and TNF-α treatment was significantly higher than group of TNF-α treatment (0.31∶0.21,t=10.49,P=0.007).Conclusion Up-regulated PC mRNA expression induced by TNF-α is significantly associated with the inhibition of HBV replication.

Adult ; Axilla ; Cell Transformation, Neoplastic ; pathology ; Diagnosis, Differential ; Epithelioid Cells ; metabolism ; pathology ; Fibrosarcoma ; metabolism ; pathology ; Humans ; Male ; Neurilemmoma ; metabolism ; pathology ; surgery ; Neuroblastoma ; metabolism ; pathology ; S100 Proteins ; metabolism ; Soft Tissue Neoplasms ; metabolism ; pathology ; surgery ; Vimentin ; metabolism

Objective:To evaluate the effect of shikonin on the expression of RANTES and chemotactic activity of monocyte in endometriosis.Methods:Established SCID endometriosis models and cultured U937 cells were treated by a series of concentration of shikonin.RANTES transcriptive expression was determined by Real-time PCR,and RANTES secretion was determined by ELISA.Furthermore,Chemotaxis assay in vitro was conducted to elucidate the effect of shikonin on chemotaxis of U937 cells by RANTES.Results:Shikonin improved the RANTES transcription of human endometrium transplanted to SCID mouse(P

Objective: To evaluate the cytotoxic effect of PEM? induced by HSPgp96 on anti-tumor in vitro. Methods; PEM? separated from mice induced by thioglycolate were divided into three groups randomly: Culture medium in control; LPS-induced group; HSPgp96-induced group. The production of NO, the cytotoxic effect to H22 cells and the morphologic change of PEM? were investigated separately by enzyme method, MTT assay and scanning electron microscope. Results: In vitro, HSPgp96 can increased NO production from PEM? of mice and significantly enhance the cytotoxic effect of PEM? to H22 cells as well as LPS. Conclusion: HSPgp96 can effectively induce the cytotoxic effect of PEM? on anti-tumor in which NO is one of the capital effective molecules in vitro.

Objective To evaluate the effects of ischemic preconditioning(IP) on liver function,complications and hospital stays after hepatectomy under hepatic vascular exclusion by a meta-analysis.Methods Randomized controlled trials(RCTs) were identified from PUBMED,EMBASE,the Cochrane Library,VIP,CNKI and Wanfang Data according to the inclusion and exclusion criteria.Literature screening,data extraction and quality assessment were made and the meta-analysis was processed by RevMan 4.2.2.Results Eight RCTs involving a total of 511 patients were included.The methodological quality was evaluated and all the trials were in graded B.The meta-analysis revealed that the postoperative ALT peak level(weighted mean difference=-176.37;95%CI:-320.67~-30.06;P=0.02)and postoperative complications incidence(odd ratio=0.64;95%CI: 0.41~0.98;P=0.04)were lower in IP group compared with control group,but there were no significant differences in blood loss,operating time,hepatic vascular exclusion time,postoperative AST and total bilirubin peak level,and hospital stays in both groups.Conclusions IP reduces the postoperative ALT peak level and complications incidence after hepatectomy under hepatic vascular exclusion,but there is no sufficient evidence to support that the IP can protect the liver from ischemia/reperfusion injury.

Objective To investigate the change of expression patterns of miRNA in the serum and peripheral blood mononuclear cells (PBMC) of lung cancer and its significance.Methods Clinical case control study was employed.Establish the method of microRNA(miRNA) detection by real time quantitative PCR (RT-qPCR).peripheral blood of the study subjects were collected in First affiliated hospital of SooChow University from November 2011 to September 2012.Gender and age matched subjects whose median age was 64(40-85) included 61 lung cancer cases,48 healthy control and benign lung diseases.We used quantitative RT-PCR to assess miRNA expression pattern of-miR-20a,21,-25,-29,-31,-126,-129,-145 and -205 in peripheral blood.U6 was taken as reference,and the expression of miRNA were indicated as F =2-△Ct,ACt =CtmiRNA--CtU6.F represents relative change of miRNA expression compared to U6 in the same sample.SPSS 19.0 was used as statistical software; t test was used for comparison of two sets of samples One way ANOVA was used for multiple groups' comparison,and make multiple comparison by the S-N-K method if the result with a significant difference.Pearson correlation analysis were used for the relationship between two variables,Brown-Forsythe test was used for Ct value equality testing among multiple samples.P ＜ 0.05were regarded as statistically significant.Results miR-20a (F =271.64,P ＜ 0.01),miR-21 (F =2232.51,P＜0.01),miR-205 (F=45.13,P＜0.01),miR-29a (F=19.98,P ＜0.01),miR-25 (F=313.19,P ＜ 0.01) and miR-126 (F =32.38,P ＜ 0.01) were differently expressed in the serum of lung cancer patients and healthy control or benign disease control.miR-29a,miR-25,miR-126 was down regulated in the development of malignant lung disease; miR-31 elevated in lung cancer compared with healthy control,while miR-145 fell; miR-31 expression changed with various differentiation of lung cancer (F =5.22,P ＜ 0.01)itwas significantly increased in the moderate-differentiated cancer,but decreased when distant metastasis existed (especially bone metastasis).But in PBMC paired with the serum samples above,statistically significance was shown in lung cancer and healthy control group and benign lung diseases group in miR-126 (F=690.58,P＜0.01),miR-129 (F=26.66,P＜0.01),miR-145 (F=48.57,P＜0.01),miR-205 (F=308.61,P＜0.01).miR-25 (F=218.57,P＜0.01) and miR-31 (F=48.05,P＜0.01),were down regulated in the development of malignant lung disease.miR-20a,miR-29a were elevated in lung cancer compared to healthy controls,miR-21 was up-regulated when distant metastasis existed; the expression of miR-31 in serum and PBMC was negatively correlated (r =-0.369,P ＜ 0.05).Areas under ROC curve of miR-25 (S =0.906,P ＜ 0.01) and miR-126 (S =0.969,P ＜ 0.01) were statistically different.Conclusions miRNA may contribute to several steps of metastasis,including local invasion,extravasation or initial survival at a distant site,and metastatic colonization,or can affect the prognosis of lung cancer.The detection of miRNA in lung cancer provides a new clue to the research of its chronic progress.

Objective To investigate the expression and significance of CD28,cytotoxic T-lymphocyte antigen-4 (CTLA-4),programmed death-1 (PD-1) and T cell immunoglobulin mucin-3 (Tim3) on T lymphocytes in chronic HBV-infected patients.Methods A total of 102 chronic HBV-infected patients,including 42 patients with chronic hepatitis B (CHB),30 patients with hepatitis B-induced liver cirrhosis (LC),and 30 patients with hepatocellular carcinoma (HCC),were enrolled from the First Affiliated Hospital to Soochow University during October 2012 and June 2013.Thirty healthy individuals were also enrolled as controls.Expression of CD28,CTLA-4,PD-1,Tim-3 on T lymphocytes in peripheral blood were determined by flow cytometry,and the differences among groups were analyzed using one-way ANOVA and LSD-t test.Spearman correlation test was performed to analyze the correlations of the expression of CD28,CTLA-4,PD-1,Tim-3 on T lymphocytes with HBV DNA loads,HBeAg and ALT.Results The expression of CD4 + CD28 +,CD8 + CD28 +,CD4 + CTLA-4 + in chronic HBV-infected patients were lower than those in healthy controls.CD4 + CD28 + expression in HCC group was lower than that in CHB group (t =2.373,P ＜ 0.05) ; CD8 + CD28 + expression in LC and HCC group was lower than that in CHB group (t =4.324 and 4.088,P ＜ 0.01) ; CD8 + PD-1 +,CD4 + Tim-3 + and CD8 + Tim-3 + expressions in CHB group were higher than those in LC,HCC group and healthy controls (t =3.051,3.130,3.121,3.254 and 3.723,P ＜0.01).CD8 + PD-1 + expression was positively correlated with ALT levels and HBV DNA loads (r =0.516 and 0.582,P ＜ 0.01) ; CD8 + Tim-3 + expression was also positively correlated with ALT levels andHBV DNA loads (r =0.578 and 0.556,P ＜0.01); PD-1 and Tim-3 expressions on CD8 T lymphocytes were positively correlated with each other (r =0.578,P ＜ 0.01).Conclusion The abnormal expression of the molecules on T lymphocytes in chronic HBV-infected patients is closely correlated with immune function disorder and the progression of the disease.

Objective To investigate the significance of changes in expression of co-stimulatory molecules on T lymphocytes in patients with chronic hepatitis B (CHB) infection.Methods In a casecontrol study,a total of 82 CHB cases including 50 male cases and 32 female cases (the mean age was 42.32 ±3.74) were enrolled in the First Affiliated Hospital to Soochow University from October 2012 to November 2013,together with 30 cases health control (15 male cases and 15 female cases,and the mean age was 42.32 ± 3.74).Patients were divided into three groups:40 cases of non-treated,30 cases of effective-treated and 12 cases of ineffective-treated with anti-viral drugs and immune-related therapy.The expression levels of CD28,CTLA-4,PD-1,Tim-3 on T cells subset from peripheral blood were determined by Flow Cytometry.Serum load of HBV DNA was detected by Real-time PCR and the serology markers such as HBeAg and ALT were detected by conventional methods The Kruskal-Wallis test was used to analysis groups comparison.Independent samples t-test and Mann-Whitney U test were used to two sample comparison.Spearman's rank correlation test was used to analyze the correlation.Results The expression levels of CD28,CTLA-4 and PD-1 on CD4 in health control group was the highest compared to ineffectivetreated group [404.65 (331.65-536.09) vs 277.15 (249.90-344.25) (H=29.81,P＜0.001);32.89 (29.69-39.69) vs 19.26 (11.90-20.56) (H =43.13,P ＜0.001),respectively].The expression level of PD-1 on CD8 in pre-treated group was the highest,while expression level in health control group was the lowest [15.47 (12.50-17.78) vs 3.05 (1.41-3.97) (H=56.60,P＜0.001)].Similarly,the expression level of Tim-3 on CD4 in ineffective-treated group was the highest,while Tim expression on CD4 in health control group was the lowest [199.62 (55.61-239.45) vs 70.62 (53.88-112.32) (H =41.03,P ＜ 0.001)].The expression level of Tim-3 on CD8 in pre-treated group was the highest,while it was the lowest in health control group [82.50 (78.69-84.58) vs 3.07 (1.56-6.87) (H=74.84,P ＜0.001)].Compared to the group with low viral load,the expression level of PD-1 on CD8 both in the pre-treated group with high viral load and the post-treated group was significantly increased [17.87 (13.38-20.94) (U=25.00,P＜0.001); 16.95 (5.39-18.27) (U=63.50,P＜0.001),respectively].Importantly,in the post-treated group,the PD-1 expression on CD4 T (r =0.689,P ＜0.001) and on CD8 T (r =0.751,P ＜ 0.001) was also positively correlated with ALT.Conclusions The abnormal expression of these co-stimulatory molecules may be present in the antiviral treatment process of CHB.It may provide new clues for the reasonable treatment of CHB.