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.

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.

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.

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.

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 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 To investigate the effects of microRNA(miRNA)-375 on gastric cancer cell function and the possible mechanism.Methods miRNA-375 eukaryotic expression vector was stably transfected into human gastric cancer cell line HGC-27.A miRNA-375 highly expressed gastric cancer cell line PC3,1-375 was screened and established.Blank plasmid transfection group (PC3.1 group) and blank control group were also established.The cell proliferation ability and apoptosis rate in three groups were detected by cell counting kit-8 (CCK-8) method and flow cytometry.The protein expression of astrocyte elevated gene 1 (AEG-1) in three groups was determined by Western blotting.The nude mouse subcutaneous xenografts model was also established.After 30 days,the nude mice were executed and the xenografts were dissociated and measured.The expression ofAEG-1 in the tumors at protein level was tested by immunohistochemistry method.The statistical inference includes t-test and analysis of variance.Results After stable transfection and cell screening,compared with that of blank control group,the expression of miRNA-375 in PC3.1-375 group significantly increased (t=2.82,2.01,bothP＜0.05).The cell proliferation of PC3.1-375 group was significantly lower than that of PC3.1 group and blank control group and the difference reached a maximum in day 3 (t3d=3.56,2.79,both P＜0.05).But with the apoptosis rate of PC3.1-375 group,the opposite was true (t=2.05,2.96,both P＜0.05).The results of Western blot shows that the expression of AEG-1 at protein level of PC3.1-375 group decreased compared with that of blank control group and PC3.1 group.The xenografts growth rate,average weight and tumor volume of PC3.1-375 group were significantly lower than those of blank control group and PC3.1 group (t=3.78,2.26,both P＜0.05).By the end of experiment,the volume of nude mouse subcutaneous xenografts in blank control group ((3.12 ± 0.77) cm3) and PC3.1 group ((3.76 ± 0.65) cm3) was significant bigger than that inPC3.1-375 group (0.72±0.21) cm3,t=5.67,6.92,both P＜0.05).The average weight of xenografts in PC3.1-375 group ((0.85 ± 0.15)g) was obviously lower than that in PC3.1 group ((4.01±0.52) g) and blank control group ((3.86±0.56) g,t=4.56,5.01,both P＜0.05).The results of immunohistochemistry indicated that the expression of AEG-1 protein in xenografts in PC3.1-375 group was remarkably decreased compared with that in blank control group and PC3.1 group (F=35.000,P＜0.05).Conclusions miRNA-375 can effectively inhibit the cell proliferation of gastric cancer,promote cells apoptosis and it's a tumor suppression factor in gastric cancer.AEG-1 may be a target gene of miRNA-375.

Objective To explore the value of serum level of interleukin 6 (IL-6),tumor necrosis factor alpha (TNF-α) and high mobility group box-1 protein( HMGB1) in early assessment the severity and prognosis of acute pancreatitis (AP). Methods Thirty-three severe acute pancreatitis (SAP) patients and 38 mild acute pancreatitis (MAP) patients were selected as study objects;and 51healthy individuals were set as control group. Serum IL-6,TNF-α and HMGB1 concentrations were determined by enzyme-linked immunosorbent assay (ELISA),the association of them and the scores of Ranson,APACHE Ⅱ . Balthazar CT,serum biochemical parameters and prognosis was analyzed.Results The serum IL-6 levels of SAP group,MAP group and healthy control group were (553. 72±175.76) pg/ml,(265. 73±179. 95) pg/ml and (16. 43±3. 32) pg/ml;and there were statistical significance of these three groups (all P＜0. 01). There was no significant difference of TNF-α in the three groups (all P＞0. 05). The serum HMGB1 levels of SAP group,MAP group and healthy control group were (11. 48±6. 94)μg/L,(6. 13±5. 80)μg/L and (1. 82±0. 64)μg/L respectively,and there were statistical significant of these three groups (all P＜0. 05). The correlation coefficient of serum HMGB1 with IL-6 and TNF-α were 0. 896 and 0. 724 (P＜0. 01) respectively. The IL-6 level was positively correlated with the scores of Ranson,APACHE Ⅱ and Balthazar CT. The TNF-αconcentration was positively correlated with APACHE Ⅱ score,and the HMGB1 concentration positively correlated with scores of Ranson and Balthazar CT. The levels of IL-6,TNF-α and HMGB1were all positively correlated with the serum creatinine concentration. The IL-6 levels of patients with local and/or systemic complications were significantly higher than those without complications.Conclusion The serum levels of IL-6,TNF-α and HMGB1 are significantly correlated with the severity of pancreatitis,all of them take part in the development of acute renal insufficiency. The high level of serum IL-6 significantly correlated with complications.

Objective To investigate the protective effects of thalidomide on rats with acute necrotizing pancreatitis (ANP) and its mechanism. Methods Fifty four SD rats were randomly divided into the three groups; ANP group, thalidomide group and control group with 18 rats in each group. The model of ANP was induced by retrograde injection of 5% sodium taurocholate into the bili-pancreatic duct. Rats in thalidomide group received thalidomide 200 mg/kg body weight gastric lavage 1 h after ANP induction. The rats were sacrificed 3 h, 6 h, and 12 h after ANP induction, and the amount of intraperitoneal ascites was quantified. The serum levels of TNF-α, IL-6, IL-18 were measured by EUSA. The proportion of CD4 + T cell, CD8 + T cell in peripheral blood was determined by flow cytometry. The expression of TNF-α mRNA in pancreatic tissue were measured by RT-PCR. The expression of ICAM-1 protein in pancreatic tissue was measured by immunohistochemistry. Pancreatic tissue underwent pathologic examination. Results Six hours after surgery, the amount of ascite, serum levels of TNF-α, IL-6, IL-18, CD4 + T cell, CD,+ T cell, pancreatic TNF-α mRNA and ICAM-1 protein expression, pathologic score in control group was (1.03 ±0.31)ml,(57.17±11.29)pg/ml, (24.45 ±4.14)pg/ml, (64.23 ±21.85)pg/ml, (47.58 ±9.21)% , (40.88 ± 2.96)%, 0.07 ±0.02, 0.57 ±0.30, 0.67 ±0.81, respectively, and the corresponding values were (3.63 ± 0.38)ml, (107.54 ±33.05) pg/ml, (47.30 ± 11.40) pg/ml, (367.76 ± 108.43 ) pg/ml, (54.90 ± 7.15)%, (17.17 ±3.12)%, 0.65 ±0.26, 3.20 ±0.57, 11.50 ±1.87 in ANP group; and (1.45 ±0.53)ml, (80.60 ±20.48) pg/ml, (26.61 ±10.85) pg/ml, (321.82 ±85.20) pg/ml, (29.80 ±2.19)% , (15.52± 1.96)%, 0.35 ±0.23, 2.37 ±0.67, 8.00 ±3.03. Besides the value of CD8 + T cell was significantly decreased, all other values were significantly increased when compared with control group (P <0.05). Conclusions Thalidomide can decrease the release of inflammatory mediator, and reduce the pathological damage of pancreas of ANP rats by inhibiting TNF-α mRNA expression.