Objective To investigate the effect of JNK signal pathway in acute necrotizing pancreatitis (ANP) associated lung injury.Methods A total of 24 SD rats were randomly divided into sham group and ANP group,ANP rats were induced by retrograde injection of 5％ sodium taurocholate into common bile duct.The rats were sacrificed 12 and 24h later,and the pancreas and lung tissue were resected and underwent routine pathologic examination,ELISA method was used to detect the level of TNF-α and IL-1β in lung tissue.Expression of JNK mRNA was detected by real time PCR,and the expression of JNK protein were evaluated by Western blotting.Results There was bleeding,necrosis,large amount of inflammatory cells infiltration in pancreatic tissue; and there was edema,pulmonary consolidation,interstitial edema,inflammatory cells infihration in lung tissue in ANP group.The levels of TNF-α and IL-1β,JNK mRNA,JNK protein,phosphorylation JNK were ( 374.3 ± 124.0) pg/ml,(649.0 ± 114.9) pg/ml,2.57 ± 0.76,1.40 ± 0.81,0.81 ± 0.20 in ANP group,which were significantly higher than those in with sham group[( 218.2 ± 68.4)pg/ml,(524.3±58.4)pg/ml,1.03±0.11,0.32±0.11,0.32±0.11,P＜0.05].Conclusions JNK signal pathway plays an important role in experimental ANP associated lung injury in rats.

Objective To investigate the relationship between the genetic polymorphisms of glutathione S-Transferase M1,T1 and the susceptibility to sporadic colorectal adenocarcinoma(SCRAC) and smoking and alcohol consumption in Chinese Hans.Methods Multiplex polymerase chain reaction (M-PCR) was used in the study of genetic polymorphisms of GSTM 1,T1 gene.Logistic analysis was performed to elucidate the roles of GSTM1,GST1,smoking and alcohol.Results The null GSTM1,T1 genotypes could increase the susceptibility to SCRAC(OR=1.711,95% CI:1.043～2.805;OR=1.734,95% CI:1.057～2.843),but smoking and alcohol consumption made no significant effect on SCRAC(OR=0.584,95% CI:0.356～0.958;OR=0.378.95% CI:0.217～0.657).Further stratification of the SCRAC patients by chnical features showed that there were no relationship between the GST M1,T1 genotype and the age of the SCRAC patients.But the frequency of null GSTM1 genotype was significantly associated with distal colon adenocarcinoma (P=0.021),colorectal adenocarcinoma of Dukes C classification (P=0.003) and poor difierentiation (P=0.020),respectively.The frequency of null GSTF1 genotype was only higher in colorectal adenocarcinoma of Dukes C classification(P=0.041).No relationship was found between the location,the degree of differentiation and the frequency of null GSTF1 genotype(P＞0.05).Furthermore,the frequencies of homozygous deletion in GSTM1,T1 genes were found to be significantly increased in SCRAC patients than those in healthy controls(38.9%VS25.7%.P=0.023).Conclusion The GST genotype is strongly correlated with SCRAC incidence in Chinese Hans.The null GSTM1,T1 genotypes can enhance the genetic susceptibility to SCRAC.while smoking and alcohol consumption have no significant effect on the susceptibility to SCRAC.

Chromatography, Reverse-Phase ; methods ; Humans ; Sorbic Acid ; analogs & derivatives ; analysis ; Urinalysis ; methods

Sugarcane bagasse modified by polyethylenimine (PEI) and glutaraldehyde (GA) was used as a carrier to immobilize Clostridium acetobutylicum XY16 in the process of butanol production. The effects of chemically modified sugarcane bagasse on batch and repeat-batch fermentations were investigated. Batch fermentation was conducted with an addition of 10 g/L modified sugarcane bagasse and 60 g/L glucose, resulting in a high solvent concentration of 21.67 g/L and productivity of 0.60 g/(L x h) with the treatment of 4 g/L PEI and 1 g/L GA. Compared to the fermentations by free cells and immobilized cells on unmodified sugarcane bagasse, the productivity increased 130.8% and 66.7%, respectively. The fibrous-bed bioreactor also maintained a stable butanol production during repeat-batch fermentations, achieving a maximum productivity of 0.83 g/(L x h) with a high yield of 0.42 g/g.
Batch Cell Culture Techniques ; Bioreactors ; Butanols ; metabolism ; Cells, Immobilized ; Cellulose ; metabolism ; Clostridium acetobutylicum ; metabolism ; Fermentation ; Saccharum ; chemistry

OBJECTIVETo study the effects of dopamin receptors-2 (DR2) on myocardial ischemic postconditioning and explore its underlying mechanisms.

METHODSThe myocardial ischemic postconditioning (PC) model was established in cultured primary rat neonatal cardiomyocytes which were then randomly assigned in the following groups: Nomial control group, Isehemia/reperfusion (L'R) group, PC (ischemic postconditioning) group, PC + Bro (Bromocriptine, a DB2 antagonist) group, PC + Hal (Haloperidol, a DB2 repressor) and PC + Hal + Bro groups. The lactate dehydrogenase (LDH) and superoxide dismutase (SOD) activity and malondialdehyde (MDA) content in cell medium were analyzed by colorunetry. The cell ultrastructure changes were observed by transmission electron microscope. The cell apoptosis was analyzed using flowcytometiy. The protein expression level of D112 and activity of p-p38 and p-JNK were detected by Western blot.

RESULTSCompared with the nonnal control group, hR increased the protein expression level of DB2, enhanced LDH activity and MDA content, promoted cell injury and apoptosis, decreased SOD activity, up-regulated the activity of p-p38 and p-JNK. Compared with the hR group, although PC further increased the expression of DR2 protein, it decreased LDH activity and MDA content, cell injury and apoptosis, increased SOD activity, down-regulated activity of p-p38 and p-JNK. Bromocriptine treatment further enhanced PC-induced canlioprotective effect, yet Hal addition attenuated this enhancing effect exerted by bromocriptine.

CONCLUSIONThe activation of DB2 is involved in the protective effect of ischemic postconditioning on myocardial ischemia/reperfusion injury through down-regulating the activity of p-p38 and p-JNK.

Animals ; Apoptosis ; Cells, Cultured ; Ischemic Postconditioning ; JNK Mitogen-Activated Protein Kinases ; metabolism ; Myocardial Reperfusion Injury ; prevention & control ; Myocytes, Cardiac ; pathology ; Rats ; Rats, Wistar ; Receptors, Dopamine D2 ; physiology ; p38 Mitogen-Activated Protein Kinases ; metabolism

Objective: To investigate the killing activity of cytokine-induced killer (CIK) cells after incubation with autologous tumor cell lysate-pulsed dendritic cells (Ag-DC) and to evaluate the immune functions of patients, the clinical efficacy and side effect of Ag-DC in combination with CIK on hepatocellular carcinoma. Methods: Peripheral blood mononuclear cells were isolated from 24 patients with hepatocellular carcinoma, and cultured with granulocyte-macrophage colony-stimulating factor and interleukin-4 to produce dendritic cells (DCs). The DCs were pulsed with autologous tumor cell lysate. T lymphocytes from PBMC were cultured with interferon-Y (IFN-γ), IL-2, CD3-moAb, and IL-1α to prepare CIK. The killing effect of CIK on SMMC-7721 was investigated after CIK was incubated with Ag-DC. After immunotherapy with Ag-DC and CIK, immunolog-ic and clinical responses of the 24 patients were evaluated. Results: The killing effect of CIK was remarkably improved after CIK was incubated with Ag-DC. The immunotherapy with DC and CIK alleviated symptoms and improved the immune functions of the patients. Except for transient fever and chill, no remarkable adverse events were observed. Conclusion: Ag-DC in combination with CIK shows short-term efficacy on hepatocellular carcinoma through inducing specific anti-tumor immunity and can be an effective adjuvant therapy for hepatocellular carcinoma.

Objective To establish the method of identifying MRSA with Taqman-fluorescence quantitative PCR basing on mecA/nuc/fem B three gene combined detecting.Methods Taking the coagulase positive MRSA,which isolated from the clinical samples and confirmed by VITEK 2 compact microbial analyzer,as the research obj ect,designed mecA/nuc/fem B specific PCR primers and Taqman fluorescent probe by bio-software PrimerPremier 5 and Designer Beacon 7,FAM,HEX and ROX markers were used to label the fluorescent probe at 5’,and the end of 3’was labeled with BHQ1,detected by fluo-rescence quantitative PCR instrment.Results ①1 g/dl gel electrophoresis results showed that the primer’s specificity of mec A/nuc/fem B were good,and molecular weight of the amplification band consistent with the expected molecular weight and no non-specific amplification band.②Three genes were obtained specific amplification in a single tube single channel and single tube multiple channel detection in PCR,and the three gene amplification effect in a single tube single tube single chan-nel and multichannel PCR similar.Conclusion Successfully established a method of multi channel Taqman-probe fluores-cence quantitative PCR identification of MRSA,mec A/nuc/fem B combined detection can effectively differentiate coagulase negative and positive MRSA,improve the accuracy of identification.

Objective To explore the value of real-time tissue elastography (RTE) with tissue dispersion quantitative analysis technique for assessment of liver fibrosis stage.Methods 51 rats were injected 6％ thioacetamide to induce liver fibrosis model,and 9 rats were injected saline as control group.In modeling 4 weeks,8 weeks,12 weeks respectively,14 rats in group of liver fibrosis model and 3 rats in control group were randomly selected to RTE.All the rats underwent tissue dispersion quantitative analysis,to obtain 12 quantitative parameters of elasticity,which included average relative strain value (MEAN),standard deviation of relative strain value (SD),area ratio of low-strain region (％ AREA),complexity (COMP),kurtosis (KURT),skewness (SKEW),contrast (CONT),entropy (ENT),inverse difference moment (IDM),angular second moment (ASM),correlation (CORR) and liver elasticity index (LF index).Subsequently,rats were sacrificed and their livers were taken for pathology analysis.Liver fibrosis model group was divided into S0,S1,S2,S3,S4 group.The 12 quantitative parameters of elasticity were compared with each group.Results 49 rats were successfully modeled,and 42 rats were analyzed.Except COMP,KURT,CORR,the other quantitative parameters had statistically differences (P ＜ 0.05).The other 9 parameters were correlated with liver fibrosis stage.Among these parameters,MEAN,％ AREA and LF index had higher related coefficient(r =-0.831,0.882,0.866).The ROC curve was made by MEAN,LF index and ％AREA to estimate the fibrosis stage,when S≥S1,S≥S2,S≥S3,S =S4,the areas under the ROCcurve were 0.884,0.925,0.934,0.962 (MEAN);0.917,0.958,0.984,0.962 (％AREA);0.917,0.948,0.966,0.967 (LF index),respectively.Conclusions As a non-invasive examination,RTE dispersion quantitative analysis technology can be used to quantitatively assess liver fibrosis.

Objective To investigate the relationship of CD4+ and CD8+ T cells with melanocytes in skin of patients with psoriasis,and to study their clinical significance.Methods Tissue specimens were obtained from both lesional and nonlesional skin of 29 patients with progressive psoriasis and 5 patients with regressive psoriasis,as well as from normal skin of 6 healthy individuals.Immunohistochemical staining was performed to determine the quantity and distribution of CD4+ T and CD8+ T cells,as well as the quantity of melanocytes and proportion of cells containing pigment granules in the basal layer of these specimens.Statistical analysis was carried out with the software SPSS 18.0 by one-way analysis of variance (ANOVA),least significant difference (LSD) test and Pearson correlation analysis.Results In patients with psoriasis,the mean number of CD4+ T cells per high-power (× 200) field was significantly larger in lesional skin than in nonlesional skin (epidermis:5.29 ± 4.66 vs.0,P＜ 0.05;dermis:77.50 ± 43.66 vs.9.67 ± 7.73,P＜ 0.05),so was the mean number of CD8+ T cells per high-power (× 200) field (epidermis:7.83 ± 6.27 vs.0.71 ± 1.20,P＜ 0.05;dermis:46.08 ± 34.26 vs.5.54 ± 4.43,P ＜ 0.05).A significant increase was also observed in the number of CD4+ and CD8+ T cells in lesional skin of patients with psoriasis compared with the normal control skin (both P ＜ 0.05).The lesional skin of patients with psoriasis also showed significandy increased number of melanocytes (103.45 ± 16.96),but decreased proportion of pigment granule-containing cells (7.45％ ± 3.86％) in the basal layer compared with nonlesional skin (43.62 ± 14.20,P＜ 0.05;43.10％ ± 14.91％,P＜ 0.05) and normal control skin (43.33 ± 14.02,P＜ 0.05;54.17％ ± 29.40％,P ＜ 0.05).There were no significant differences in either the mean number of CD4+ T cells,CD8+ T cells and melanocytes or the proportion of pigment granule-containing cells between nonlesional psoriatic skin and normal control skin (all P ＞ 0.05).The mean number of melanocytes was significantly higher in regressive psoriatic lesions than in white patches arising in subsided psoriatic lesions (P ＜ 0.05) and normal control skin (P ＜ 0.05),but similar between white patches and normal control skin (P ＞ 0.05),while the proportion of pigment granule-containing cells was insignificantly lower in regressive psoriatic lesions than in white patches (P ＞ 0.05),and significantly lower in regressive psoriatic lesions and white patches than in normal control skin (both P ＜ 0.05).Neither the number of CD4+ T cells nor that of CD8 + T cells was correlated with the number of melanocytes or the proportion of pigment granule-containing cells in progressive psoriatic lesions (both P ＞ 0.05),while the number of both CD4 + T cells and CD8 + T cells was positively correlated with that of melanocytes (r =0.46 and 0.56,respectively,both P ＜ 0.05),but uncorrelated with the proportion of pigment granule-containing cells in nonlesional psoriatic skin (both P ＞ 0.05).Conclusions In progressive psoriatic lesions,there is a significant increase in the number of CD4+ and CD8 + T cells as well as melanocytes in the basal layer,but a significant decrease in the proportion of pigment granule-containing cells.After subsidence of psoriatic lesions,both the number of melanocytes and proportion of pigment granule-containing cells gradually reach the levels in normal skin of healthy individuals.

Objective To evaluate the effect of small-dose ketamine on the onset time and course of modified electroconvulsive therapy (MECT) in mentally depressed rats.Methods Sixty SPF adult male SpragueDawley rats,aged 2-3 months,weighing 220-250 g,were randomly divided into 6 groups (n =10 each) using a random number table:normal control group (group C),depression group (group D),ECT group,propofol + ECT group (group PE),ketamine + ECT group (group KE) and ketamine + propofol + ECT group (group KPE).The depression model was established by chronic unpredictable mild stress (CUMS).Mter CUMS,C,D and ECT groups received intraperitoneal normal saline 8 ml/kg,group PE received intraperitoneal propofol 100 ml/kg,group KE received intraperitoneal ketamine 10 ml/kg,and group KPE received intraperitoneal ketamine 10 ml/kg + propofol 80 ml/kg.All the groups received ECT once a day for 7 consecutive days starting from the time point when righting reflex was lost except C and D groups.Open-field test was performed before CUMS,at 1 day after CUMS and at the end of each ECT (T0 8).The total distance and the number of standing on the back legs were recorded.Morris water maze test was performed at 2 days after CUMS and 1 day after the end of therapy,and the escape latency and time of staying at the original platform quadrant were recorded.Results Compared with group C,the total distance was shortened and the number of standing on the back legs was reduced,the escape latency was prolonged,and the time of staying at the original platform quadrant was shortened at T1-8 in D,ECT,PE and KE groups and at T1 5 in KPE group,and no significant was found in KPE group in the total distance,number of standing on the back legs,escape latency,and time of staying at the original platform quadrant at T6-8.Compared with group D,the total distance was prolonged and the number of standing on the back legs was increased at T6-8 in ECT and PE groups and at T4-8 in KE and KPE groups,the escape latency was prolonged,and the time of staying at the original platform quadrant was shortened in ECT group,and the escape latency was shortened,and the time of staying at the original platform quadrant was prolonged in KPE group.Compared with ECT and PE groups,the total distance was prolonged and the number of standing on the back legs was increased at T4-7 in group KE and at T4-8 in group KPE,and the escape latency was shortened,and the time of staying at the original platform quadrant was prolonged in KPE group.Compared with group KE,the total distance was prolonged and the number of standing on the back legs was increased at T6.7,the escape latency was shortened,and the time of staying at the original platform quadrant was prolonged in KPE group.Conclusion Small-dose ketamine can shorten the onset time and course of MECT in mentally depressed rats.