ObjectiveTo investigate the expression characteristics of Caspase-8 in gastric carcinoma tissues and its correlation with clinical pathological features.Methods The mRNA and protein of Caspase-8 in gastric carcinoma and its surrounding tissues were detected by reverse transcription polymerase chain reaction (RT-PCR) and immunohistochemistry,respectively.ResultsThe Caspase-8 mRNA in gastric carcinoma tissues was obviously lower than that in its surrounding ones (0.154±0.065 vs 0.394±0.107,t =14.04,P ＜0.01),which was correlated with lymph node metastasis,tumor stage,differentiation grade (P＜0.01).The Caspase-8 protein in gastric carcinoma tissues was also significantly lower than that in its surrounding ones (31.48％ vs 85.19％,x2 =32.04,P ＜ 0.01),which was correlated with tumor stage,differentiation grade (P＜0.05).ConclusionsCaspase-8 is low-expressed in gastric carcinoma,which may play an important role in the occurrence and progression of gastric carcinoma.

Background It is well known that tissue factor (TF) is expressed in tumor cells and neovascular endothelial cells of tumor.It plays an important role in the formation of new blood vessels as well as the growth and metastasis of tumor.However,whether TF is expressed or not in choroidal melanoma(CM)is unclear.Objective This study is to investigate the expression of TF in a choroidal melanoma cell line and human choroidal melanoma.Methods The expression of TF was studied in the optimal choroidal melanoma-1 (OCM-1) cell line and ten specimens from CM patients using immunhistochemistry.Ten normal eye specimens from donators were used as controls.Results The TF protein was mainly expressed in the cytoplasm.It is over-expressed in OCM-1 cells with positive rate of 85.33±5.47%.Hyper-expression of TF also was found in human choroidal melanoma with a positive rate of 41.60±14.17%.The integrated optical density (IOD) of positive cells in CM was 33853.67±16445.30,and only 5.65±4.26% of positive cells was found in normal human choroidal tissue.The IOD of TF expression in normal human choroidal tissue was 426.43±316.62.Conclusion The overexpression of TF in CM cells may be a new immunotherapy target for CM treatment.

Objective To study the effect of Pingl? Formula (PL) on the arrhythmia of myocardial ischemia-reperfusion in rats.Methods Rat arrhythmia models were established by occlusion of the LAD coronary artery for 15 min and thereafter reperfusion for 45 min.At the same time,the changes were observed on lead Ⅱ dynamic ECG.The activities of Ca~(2+)-ATPase,Mg~(2+)-ATPase,and Na~+,K~+-ATPase were determined, respectively.The levels of ATP,total adenine nucleotides(TAN),and energy charge(EC) of the myocardial organized energy state were analyzed by RP-HPLC.Results PL(ig 0.04,0.20,and 1.00 g/kg) showed to decrease the incidence of arrhythmia induced by ischemia-reperfusion,shorten the duration of ventricular arrhythmia and eliminate the incidence of ventricular fibrillation.The activities of Ca~(2+)-ATPase,Mg~(2+)-ATPase,and Na~+,K~+-ATPase were increased by PL.In the PL(0.04,0.20,and 1.00 g/kg) groups,ATP levels recovered to 58%,68%,and 86% as compared with normal group,simul-taneously,TAN values were enhanced to 25%,49%,and 66% and EC values to 24%,27%,and 29%, respectively.Conclusion PL has obvious antiarrhythmic effect on rat,the protective action is related to the improvement of energy metabolism.

In order to develop an anti-FMDV A Type monoclonal antibo by (mAb),BABL/c mice were immunized with FMDV A type.Monoclonal antibodies (mAbs) 7B11 and 8H4 against Foot-and-mouth disease virus (FMDV) serotype A were produced by fusing SP2/O myeloma cells with splenocyte from the mouse immunized with A/AV88.The microneutralization titer of the mAbs 7B11 and 8H4 were 1024 and 512,respectively.Both mAbs contain kappa light chains,the mAbs were IgG1.In order to define the mAbs binding epitopes,the reactivity of these mAbs against A Type FMDV,were examined using indirect ELISA,the result showed that both mAbs reacted with A Type FMDV.These mAbs may be used for further vaccine studies,diagnostic methods,prophylaxis,etiological and immunological research on FMDV.Characterization of these ncindicated that prepared anti-FMDV A mAbs had no cross-reactivity with Swine Vesicular Disease (SVD) or FMDV O,Asial and C Type antigens.Their titers in abdomen liquor were 1:5×106 and 1:2×106,respectively.7B11 was found to be of subtype IgG1,8H4 was classified as IgG2b subtype.The mAbs prepared in this study,are specific for detection of FMDV serotype A,and is potentially useful for pen-side diagnosis.

Objective To investigate the diagnostic value of a quantitative detection of K-ras mutation in samples from endoscopic ultrasound-guided fine-needle aspiration (EUS-FNA)of pancreatic cancer.Methods Samples taken by EUS-FNA from 53 pancreatic occupying lesions were collected, and the copies of wild-type and mutated K-ras gene was measured by PNA-clamping real-time quantitative PCR. The results were analyzed with refer to cytological findings to evaluate its clinical values. Results According to cytological finding, a total of 37 cases were diagnosed as pancreatic cancer, and 16 were non-malignant lesions. Kras mutation was detected in 83.8% of cancer cases, and 18. 8% of non-cancer cases, which was significantly different ( P ＜0. 05 ). Sensitivities of cytology and K-ras examination were 59. 5% and 83.8%, respectively, while that of combination of cytology and K-ras examination was 89. 2%. Conclusion Quantitative analysis of the mutant K-ras gene in samples taken by EUS-FNA is a useful tool for diagnosing the pancreatic carcinoma.

Objective To investigate the diagnostic value of intravoxel incoherent motion diffusion weighted imaging (IVIM-DWI) and dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI)in differentiating non-metastatic from metastatic mesorectal lymph nodes in rectal cancer.Methods IVIM-DWI and DCE-MRI were performed preoperatively in 38 patients with histologically proven rectal carcinoma.The short axis diameter,short-to-long axis diameter ratio,four IVIM-based parameters (ADC,D,D? and f) and six DEC-MRI semi-quantitative parameters (Slope,Maxslope,CER,Washout,TTP,iAUC90 and iAUC180 )were compared between the metastatic (n=28)and non-metastatic (n=27)lymph nodes.Results There were significant statistical significances between the metastatic and non-metastatic lymph nodes in mean short axis diameter (8.87 mm±2.829 mm vs 6.83 mm±1.075 mm),D value[(0.824±0.1 13)× 10 -3 mm2/s vs (1.033±0.244)× 10 -3 mm2/s],CER(1.588 ±0.664 vs 1.054 ±0.41 9),iAUC90 (22.89 ± 9.83 vs 13.59 ± 5.34)and iAUC1 80 (49.38±20.1 9 vs 30.31 ± 1 1.67)(P ≤0.001).The short-to-long axis diameter ratio,ADC,D? ,f,Slope,Maxslope,Washout and TTP values did not show significant differences between the two groups(P >0.05).The respectively optimal cut-off value (area under the curve,sensitivity and specificity)for distinguishing metastatic from non-metastatic lymph nodes were as follows:short axis diameter=7.1 mm(0.744,64.2%,85.1%),D=0.906×10 -3 mm2/s (0.821,81.5%,75.0%),CER=1.05(0.749,85.7%,62.9%), iAUC90 =13.42(0.780,85.7%,62.9%),iAUC180 =49.65 (0.770, 50.0%,100%)respecyively.Conclusion Both IVIM-DWI and DCE-MRI are useful for differentiating non-metastatic from metastatic mesorectal lymph nodes in rectal cancer.

Objective To determine the serum levels of miR-16,miR-126 and miR-221 in type 2 diabetes (T2DM)patients with or without microvascular complications,and further evaluate their clinical significance.Methods The serum levels of miR-16,miR-126 and miR-221were examined in 55 T2DM patients,55 T2DM patients with microvascular complications and 5 5 healthy controls using quantitative real-time PCR (qRT-PCR).The levels of fasting blood glucose,triglycerides,choles-terol,high density lipoprotein,low density lipoprotein and others biochemical parameters were determined by biochemical an-alyzer,and the diagnostic usefulness of the three miRNAs for T2DM patients and patients with microvascular complications were assessed by ROC curve analysis and logistic regression analysis.Results Compared with healthy controls [miR-1 6 (14.35±1.00)×10-5 ,miR-126(11.75±1.47)×10-5 and miR-221(32.26±3.98)×10-5 ],the miR-16,miR-126 and miR-221 expression were significantly increased in T2DM patients [miR-16(23.74±2.70)×10-5,miR-126(25.01±4.13)× 10-5 and miR-221(84.76±11.79)×10-5 ]and T2DM patients with microvascular complications [miR-16(43.74±9.61)× 10-5 ,miR-126(17.66±2.20)×10-5 and miR-221(82.52±12.48)×10-5 ].The area under ROC curve (AUCROC)of miR-16,miR-126 and miR-221 for T2DM patients were 0.63 (95%CI 0.53~0.74),0.64 (95%CI 0.54~0.74)and 0.74 (95%CI 0.65~0.83),respectively.For T2DM patients with microvascular complications,the area under ROC curve were 0.75 (95%CI 0.66~0.84),0.62 (95%CI 0.52~0.73)and 0.73 (95%CI 0.64~0.83),respectively.Furthermore,logistic re-gression revealed that the three miRNAs were novel independent risk factors for T2DM and T2DMC.Conclusion The levels of miR-16,miR-126 and miR-221 were significantly increased in the serum of T2DM patients with or without microvascular complications,and can be used as potential non-invasive biomarkers and risk factors for T2DM patients and T2DM patients with microvascular complications.

Objective To explore the characterization of the long-term survival patients through monitoring the lymphoeytes subgroup and cytokines level.Method The lymphocytes subgroups (T,B,and NK) and cytokines (IL-9,-17,-22) of patients and healthy groups were tested by using flow cytometry and ELISA.Results The levels of CD3+,CD4+,CD8+,CD8+ CD28+/CD8+ and IL-9 were gradually increased in the short-term group and long-term group as compared those in healthy group.The percentage of CD3 + cells and level of IL-9 were significantly lower in short-term group and long-term group than in healthy group (P＜0.05).The percentage of CD8+ cell was significantly lower in short-term group than in healthy group (P＜0.05).The ratio of CD8+ CD28+/CD8+ was significantly lower in short-term group than in longterm group and healthy group (P＜0.05).The percentage of B cells and NK cells,and IL-22 were gradually increased in the healthy group,shortterm group and long-term group.The percentage of B cells and NK cells was significamly higher in long-term group than in healthy group (P＜0.05).The level of IL-22 was significantly higher in longterm group than in short-term group and healthy group (P＜0.05).However,NKT lymnphocytes and IL-17 showed no statistically significant difference between long-term group and short-term group.Conclusion The ratio of T lymphocyte subgroups and the level of IL-9 were good biomarkers for evaluating the immune characterization of OLT patients; NK and B lymphocytes,and IL-22 may be associated with the long-term survival in patients after OLT.

ObjectiveTo detect the Alu expression in the stool of patients with pancreatic cancer and investigate its value in the diagnosis of pancreatic cancer.MethodsStool samples were obtained from patients with pancreatic cancer (PC) ( n =41 ),chronic pancreatitis (CP) ( n =27 ) and healthy subjects ( n =23 ),the DNA was extracted from the stool and the expression of Alu repetitive sequences was subjected to quantitative analysis by the real-time PCR.ResultsThe expressions of Alu repetitive sequences in PC,CP,and healthy subjects were (5.17 ± 0.99 ),( 3.79 ± 0.94),(0.28 ± 0.35 ) rig/g,and the difference among the three groups was statistically significant (P ＜0.05).The AUC of PC was 74.8％ with the 95％ CI 0.661 ～0.835,and the sensitivity,specificity was 75.6％ and 67.1％,respectively.ConclusionsAlu repetitive sequences are highly expressed in the stool of patients with pancreatic cancer,and it is of value in the diagnosis of pancreatic cancer.

Objective To use hemin as a catalyst in the formation of disulfide bonds in the synthesis of linaclotide. Methods The linaclotide peptide was synthesized by the standard 9-fluorenylmethyl(Fmoc)solid-phase synthetic strategy. Wang resin and Trt-protected cysteine were used in the synthesis. Hemin was used in random oxidation of line linaclotide. The result was compared with those of air,dimethyl sulfoxide(DMSO),and I2 oxidation systems. Results and Conclusion Hemin is a highly effective catalyst for disulfide bond formation in linaclotide synthesis. It overcomes some disadvantages in oxidation reactions with conventional oxidative re-agents,and supplies a convenient way for the synthesis of peptide with concentrated disulfide bonds.