Objective To investigate the effect of VIP on gut barrier function in rats with acute neerotizing panereatitis (ANP). Methods Fifty four SD rats were randomly divided into three groups: sham operated (SO) group,ANP group and VIP group. Each group was subdivided into 1h, 6h and 12h subgroup after the models were established and each subgroup had 6 rats. The models of ANP were indueed by retrograde injection of 4% sodium taurocholate into the pancreatic duet. VIP group was induced by 5 nmol VIP intraperitoneal injection within 5 minutes after the establishment of ANP model. The serum VIP and intestinal homogenate VIP were detected with ELISA. The serum endotoxin was tested by MB-80 microbes dynamic detecting system. The expression of TNF-α, IL-6, IL-10 mRNA in gut mncosa were determined by RT-PCR.Intestinal samples were harvested for pathological examination. Results The intestinal structure was significantly damaged in ANP group, and the extent of pathological changes were ameliorated in VIP group.The serum and intestinal homogenate VIP levels 6h after the establishment of ANP model were (49. 582 ±3. 735) pg/ml and (87. 731 ±4.601 ) pg/g pro, respectively, which were significantly lower than that of SO group (P ＜ 0.05 ). 12 h after the establishment of SAP model, the serum and intestinal homogenate VIP levels in ANP group were (65. 192 ± 5. 785) pg/ml and ( 110. 978 ± 6. 420) pg/g pro, respectively, which were significantly higher than that of SO group. The serum endotoxin, expression of TNF-α, IL-6, IL-10 mRNA in gut mucosa were(29.570 4-5.127)pg/ml,0.861 ±0.081,1.150 ±0.187 and 0.786±O.102,respectively,which were significantly higher than those of SO group(P＜0.05).The serum endotoxin,expression of TNF-α,IL-6 in gut mucosa in VIP group 6h after establishment of ANP model were(20.486 ±2.81 1)pg/ml,0.707±0.095 and 0.889±0.136,respectively,which were significantly lower than those of ANP group(P＜0.05).The expression of IL-10 mRNA in VIP group was 0.992 ±0.126,which was significantly higher than that of ANP group(P＜0.05).Conclusions VIP had significant protective effects on gut barrier function in rats with ANP.

Objective To investigate the protective effects of vasoactive intestinal peptide(VIP) on gut barrier in rats with severe acute pancrititis(SAP)and its mechanisms.Methods Fifty-four SD rats were randomly divided into sham operated(So)group,SAP group and VIP group.The model of SAP was induced by retrograde injection of 4%sodium taurocholate into the bili-pancreatic duct.The rats in VIP group were intraperitoneal injected with VIP of 5×10-9 nmol 5 minutes after model established.The endotoxin in plasma was measured at 1,6 and 1 2 hour.The expression of Toll-like receptor(TLR)4 in gut mucosa was examined using RT-PCR and immunohistochemistry and mucosa tissue were detected using electroscopic examination.Results The concentration of endotoxin in plasma and expression of TLR4 were both increased at 1 hour and reached peak at 1 2 hour in SAP group compared with SO group(no expression of TLR4),but were decreased in VIP group.A correlation was found between endot6xin and expression of TLR4.The electroscopic examination showed that the pathological injury in intestine was severe in SAP group than that in VIP group.Conclusion The protective effect of VIP in gut barrier function may contribute tO down-regulation of TLR4 expression in gut mucosa.

Background and purpose: At present, gastric cancer is considered to be both genetic and epigenetic disease, and epigenetic alterations play a significant role in the development of gastric cancer. DNA methylation is the most well studied and most in-depth epigenetic modiifcations in human-beings. The silencing of tumor-related genes by DNA methylation is reversible. ERCC1 is a kind of DNA repair gene. The present study was aimed to detect the CpG island methylation status of ERCC1 gene promoter in gastric cancer tissues and corresponding peripheral blood, and to explore the relationship between methylation of ERCC1 gene in peripheral blood and in gastric cancer tissues. Methods:Methylation speciifc PCR was performed to detect the methylation status of ERCC1 gene in the tumor tissues and the paired peripheral blood from 30 gastric cancer patients. Results:The positive rate of methylation of ERCC1 gene promoter CpG island was 76.7%(23/30) in the tumor tissues and 63.3%(19/30) in serum of gastric cancer patients, and the difference had no statistical signiifcance. Conclusion:Our studies suggest that ERCC1 gene promoter CpG island methylation can be detected in a high proportion of the serum consisting with that in tumor tissues of gastric cancer patients, and the detection of methylation status of ERCC1 gene in peripheral blood provides a more simple, fast and reliable way for the medical treatment of gastric cancer and also provides the possible theoretical basis for the CpG island methylation of ERCC1 gene promoter as a target for the treatment of gastric cancer.

Objective:This study aims to detect the CpG island methylation status of the ERCC1 gene promoter and the expres-sion of the ERCC1 protein in gastric cancer tissues, as well as to investigate the correlation and significance between ERCC1 gene pro-moter CpG island methylation and protein expression. Methods:Methylation-specific PCR was performed to detect the methylation sta-tus of the ERCC1 gene in tumor tissues and adjacent cancerous tissues from 30 gastric cancer patients. Ten cases of normal gastric tis-sues were used as control. Expression of the ERCC1 protein in gastric cancer tissues, adjacent cancerous tissues, and normal gastric tis-sues was examined by immunohistochemistry S-P method. Results:The methylation rate of the ERCC1 gene promoter CpG island in gastric cancer tissues was significantly higher than that in adjacent cancerous tissues (76.7%vs. 13.3%, P<0.05), and the difference was statistically significant. The incidence of promoter methylation was not found in 10 normal gastric tissues. The negative rate of ERCC1 protein expression in 30 cases of gastric carcinoma tissues was significantly higher than that in adjacent cancerous tissues (70.0% vs. 33.3%, P<0.05), whereas 10 normal gastric tissues all exhibited positive expression for the ERCC1 protein. The tumor tissues with ER-CC1 gene promoter CpG island methylation showed a lower expression of ERCC1 protein than the unmethylated tumor tissues in gas-tric cancer patients. Conclusion:Methylation of the ERCC1 gene promoter CpG island and protein expression are negatively correlat-ed, and methylation of the CpG island of the ERCC1 gene may be one of the main reasons for the down-regulation of protein expres-sion.

It was widely received that the parafasicular nucleus (Pf) and the central lateral nucleus of the thalamus played an important role in the mechanism of the acupuncture analglsia. Consequently, it was necessary to make a careful histological observation on pf in order to provide some data for the investigation of the acupuncture anesthesia mechanism. Five adult rabbits, each weighing about 2 kg, were selected for this experiment. The animals were infused with 10% formalin, their skulls were mounted on the stereotaxic apparatus according to the Sawyer's atlas. Brains were removed and made serial sections which were stained alternately with Nissl's method and the silver impregnation method of Glees. The form and size of the Pf was studied with a microprojector, while the diameters of the Pf neurones were measured by a micrometer under immersion microscope. Results were as follows. The diameters of the whole Pf: from dorsal to ventral, were about 1.6～2.2 mm, from anterior to posterior, were 0.9～1.5 mm, the transverse diameters of Pf were about 0.8～1.1 mm. The volume of Pf was about 0.68～1.26 cubic mm. The total number of the neurones in Pf was about 21,000～39,000 unilaterally. The long diameter of the cell soma in Pf was 21.06?6.22?, the short diameter of the cell soma was 11.20?3.28?. The ratio of the long diameter to the short diameter of the cell soma was 1.91?0.84. The area of the cell soma was about 143.28?58.92?~2. The long diameter of the cell nucleus of Pf was 11.74?1.5?, the short diameter of the cell nucleus was 8.02?2.01?. The ratio of the long to short diameter of the cell nucleus was 1.44?0.32. The Pf of the rabbit exhibited primary differenciation. The neurones of Pf could be subdivided into three groups: (1) The first group was formed by large and deeply stained cells which were triangular or pyramidal in shape, and distributed mainly in the lateral inferior portion of the Pf. (2) The neurones of the second group were fusiform in shape, concentrated mainly in the medial portion of the Pf. (3) The neurones of the third group were smaller than those of the first group and stained more lightly, they were shortly ellipsoidal in shape and distributd chiefly in the lateral superior portion of the Pf. Statistical analysis showed that there were some differences among the cells in different portions of the Pf. The authors suggested that the large and deeply stained neurones in Pf were quite similar to those of the central lateral nucleus and might belong to pain sensitive neurones.

Background:The ability of tumor cells to evade destruction by immune system is known as tumor immune escape. Regulatory T cells( Treg cells)and myeloid-derived suppressor cells( MDSCs)are considered to be the most critical cell subsets participating in tumor immune escape. Aims:To determine the percentages of Treg cells and MDSCs in peripheral blood of patients with gastric cancer for investigating the effects of these two cell subsets on development and progression of gastric cancer. Methods:Peripheral blood specimens from 77 patients with gastric cancer and 20 healthy controls were collected for measurements of CD4+CD25+Foxp3+ Treg cells and MDSCs by flow cytometry. Correlations between Treg cells,MDSCs and clinicopathological characteristics of gastric cancer were analyzed. Results:Percentage of CD4+CD25+Foxp3 + Treg cells in peripheral blood CD4+ T cells and percentage of MDSCs in peripheral blood mononuclear cells were both significantly higher in gastric cancer patients than in healthy controls( Treg cells:4. 72% ± 1. 01% vs. 1. 57% ± 0. 99%,P<0. 01;MDSCs:21. 72% ± 10. 12% vs. 2. 90% ± 1. 80%,P<0. 01). Percentage of Treg cells in peripheral blood was correlated with the clinical stage,depth of invasion and lymph node metastasis of gastric cancer(P<0. 05), while percentage of MDSCs was correlated with the clinical stage of gastric cancer(P<0. 05). Furthermore,a significant positive correlation was observed between percentages of Treg cells and MDSCs in peripheral blood of gastric cancer patients ( rs =0. 681,P<0. 01). Conclusions:Gastric cancer patients are characterized by high expressions of immunosuppressive cells in peripheral blood,such as Treg cells and MDSCs,which might be related with the development and progression of gastric cancer via tumor immune escape.

After the initial episode of acute pancreatitis (AP), some patients have a tendency to relapse.With the development of imaging technologies and genetic tests, the diagnostic accuracy of the etiological factors of recurrent acute pancreatitis (RAP) such as dysfunction of sphincter of Oddi, pancreas divisum, and genetic mutations are improved.Clinical studies indicate that etiological treatment by endoscopic approaches may reduce recurrence in some RAP patients.In this article, the progress in etiology, diagnosis and treatment of RAP was reviewed.

Objective To introduce a novel modified CT severity index based on the assessment of extrapancreatic inflammation and pancreatic necrosis on CT index (EPIPN) and to evaluate its effect in predicting the severity and prognosis of acute pancreatitis. Methods Seventy-seven consecutive patients diagnosed as acute pancreatitis (AP) from August 2006 to December 2007 were retrospectively analyzed. The clinical data included age, sex, cause, the C-reactive protein(CRP) level with in 72 hours of onset of symptom, Ranson signs, the APACHE I1 score, the disappearing time of the abdominal pain, the presence of organ failure, the length of hospital stay, etc. All patients underwent contrast-enhanced multisection CT scan after admission of 48-72 hours. The CT severity index (CTSI) and EPIPN scores were obtained. The severity of pancreatitis for each patient was then categorized as severe if CTSI≥7 or EPIPN>5. The diagnostic value of EPIPN in predicting the severe acute pancreatitis (SAP) was compared with that of CTSI using ROC curve. The correlation of EPIPN or CTSI with clinical coutcome was conducted. Results Of 77 patients, 34 were males and 43 were females with mean age of 51.79 years (age range 22-92 years). The causes of AP were gallstones (63 cases), hyperlipemia (6 cases), alcohol (1 case) and idiopathic (7 cases). Organ system failure was present in 14 (18.2%) of the 77 patients. The area under the ROC curve of CTSI in predicting the SAP was 0.72 (95% CI: 0.59-0.88) with sensitivity of 80.4% and specificity of 55% when CTSI≥7, and that in EPIPN was 0.82 (95% CI: 0.73-0.91) with sensitivity of 91.3% and specificity of 63% when EPIPN >5. EPIPN was well correlated with hospital stay, APACHE Ⅱ score and CRP levels. Conclusions The EPIPN allows accurate estimation of disease severity and prognosis in AP patients. The diagnostic effect of EPIPN for predicting SAP is superior to CTSI. The EPIPN index is-both convenient and practical, and has clinical value.

Objective To investigate the effects of soluble CD40 ligand combined with LY294002,a specific inhibitor of phosphatidylinositol 3-kinase,on proliferation and invasion of gastric cancer cells (AGS ) and its potential mechanisms. Methods The effect of gradient concentration of sCD40L and its combination with LY294002 on cellular proliferation was determined using MTT assay. The ability of cellular invasion and migration was evaluated by wound healing and Transwell invasion assay. The protein expression of PI3K, p-Akt and vascular endothelia growth factor (VEGF) was tested by Western blotting. Results The inhibitory effect of cellular proliferation in sCD40L, LY294002 and sCD40L + LY294002 groups was 70. 1%, 65. 7% and 41. 3%, respectively, with significant difference (P<0. 05). And wound healing was also delayed in cells treated with sCD40L plus LY294002 when compared with those treated with sCD40L or LY294002. These results indicated that the inhibitory effect of cellular proliferation, migration and invasion was enhanced when cells were treated with sCD40L plus LY294002. The protein expression of PI3K,p-Akt and VEGF increased in cells treated with sCD40L, but reduced drastically in cells treated with sCD40L plus LY294002. Conclusion Inhibiting PI3K/Akt signaling pathway may enhance the inhibitory effect of proliferation, migration and invasion by sCD40L on AGS cells, which provide a new way for biochemical therapy of gastric cancer.

Objective To quantitatively detect the circulating DNA in plasma of patients with acute pancreatitis and to evaluate its potential clinical values. Methods Blood samples collected from 40 patients with mild acute pancreatitis (MAP), 20 with severe acute pancreatitis (SAP) and 50healthy controls were extracted for DNA using genomic DNA extraction kit. The DNA level was detected by real-time fluorescence quantitative PCR assay. The clinical association of DNA level with acute pancreatitis was analyzed. The receiver operating characteristic (ROC) curve was set up. Results The median concentration of plasma DNA in SAP group was 24.84 ng/ml, which was significantly higher than that in MAP group (7.60 ng/ml, P=0.006) and the healthy controls (5.23 ng/ml, P=0.000). The median concentration of plasma DNA in patients with acute pancreatitis was higher than in healthy controls (P=0.006), however, there was no significant difference between MAP group and healthy controls (P=0.322).The area under the ROC curve performed by the plasma DNA from SAP and MAP groups was 0. 881(95% CI, 0.773-0.989). With a cutoff value of 11.20 ng/ml, the overall sensitivity was 19/20% and specificity was 72.5%. Plasma DNA level was found to be associated with APACHE-Ⅱ score (P=0.001), Ranson score (P=0. 013), serum Ca2+ level (P=0.000), and c-reactive protein levels (P=0.001). Conclusions Plasma DNA is correlated with the extent of pancreatitis. It can be used in monitoring the development of acute pancreatitis and may be a potential marker for early diagnosis of SAP.