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.

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 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.

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 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 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 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.

Adenocarcinoma ; genetics ; pathology ; surgery ; Adult ; Aged ; Aged, 80 and over ; Biopsy ; methods ; Bronchoscopy ; Carcinoma, Large Cell ; genetics ; pathology ; surgery ; Carcinoma, Non-Small-Cell Lung ; genetics ; pathology ; surgery ; Carcinoma, Squamous Cell ; genetics ; pathology ; surgery ; Female ; Gene Amplification ; Humans ; Lung Neoplasms ; genetics ; pathology ; surgery ; Male ; Middle Aged ; Polyploidy ; Receptor, Epidermal Growth Factor ; genetics ; Young Adult

In this study, the relationship between mycelium morphology and laccase production was studied. The results indicated that the morphology of P. ferulae pellets was changed when glass beads were added. Laccase production showed higher with spherical mycelium than with filamentous or flocculent mycelium. In addition, the spherical mycelium with a diameter of 0.2-0.4 mm highly affected laccase production. Effect of the composition of culture medium on pellets was investigated and results indicated that various concentrations of glucose, corn meal and wheat bran were important to the formation of pellets in diameter of 0.2-0.4 mm. Besides nutrients, the addition of non-nutritional substrates influenced the distribution of P. ferulae pellets. However, the production of laccase was not promoted by non-nutritional substrates.
Culture Media ; Fermentation ; Glass ; chemistry ; Industrial Microbiology ; Laccase ; biosynthesis ; Mycelium ; cytology ; growth & development ; Pleurotus ; cytology ; enzymology

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.