Objective:To establish an LC-MS/MS method for the simultaneous determination of acetaminophen and its five metabolites in mice plasma,and investigate the metabolism of acetaminophen by using the method.Methods:Para aminobenzoic acid was used as the internal standard.The plasma samples were precipitated by methanol,and then separated on a C18 column with the mobile phase of methanol and 5 mmol·L-1 ammonium acetate buffer solution containing 0.1% formic acid (55∶45).The flow rate was 0.5 ml·min-1,and the column temperature was 25℃.An electrospray ionization source was applied and operated in a positive ion mode using MRM:APAP,-,m/z 152.0→110.0;APAP-cys,-,m/z 271.2→140.1;APAP-glut,-,m/z 457.0→328.0;APAP-NAC,-,m/z 313.4 →208.0;APAP-sulf,-,m/z 232.4→152.1;APAP-gluc,-,m/z 328.2→152.1;IS,-,m/z 138.2→120.0.Results:The method exhibited good linearity over the concentration range of 0.2-10 μg·ml-1for APAP,1.0-20 μg·ml-1 for APAP-gluc,1.0-20 μg·ml-1 for APAP-sulf,1.0-20 μg·ml-1 for APAP-glut,0.4-15 μg·ml-1 for APAP-NAC and 0.2-10 μg·ml-1 for APAP-cys (r≥0.990 0).The inter-day accuracy and precision of acetaminophen and its five metabolites were all below 15%.The average recovery was between 85% and 115%,and RSDs were all below 15%.Conclusion:The LC-MS/MS method is proved to be quick,sensitive and accurate,and suitable for the determination of acetaminophen and its five metabolites in mice plasma.

ObjectiveTo evaluate the therapeutic effect of endoscopic full-thickness resection (EFTR) for gastric stromal tumors.MethodsA total of 33 patients with gastric stromal tumor orgination from deep muscularis propria layer received EFTR from January 2010 to July 2011.The effectiveness and safety of EFTR were compared with those of other 34 patients with gastric stromal tumor origination from muscularis propria layer who underwent endoscopic submucosal dissection (ESD).ResultsExcept in 2 patients with lesions larger than 3.0 × 3.0 cm,EFTR was successful in others 31 patients,who recovered well and had no recurrence during the follow-up within 12 months.There were no significant differences in resection rate,incidence of complications,body temperature,white blood cell counts or recovery time between 2 procedures (P ＞ 0.05 ).However,the number of clips used in EFTR ( 7.0 ± 3.5 vs.4.9 ± 3.1,P =0.013 ) and postoperative fasting days (3.4 ± 1.5 vs.2.0 ± 1.0,P =0.001 ) were significantly higher than those of ESD procedures.ConclusionEFTR is effective and safe for gastric stromal tumors with no higher risk than ESD,but it is more complex technically.EFTR can be used as an expanding method of ESD in endoscopic treatment of gastric stromal tumors.

Objective To investigate the regional distribution and morphological features of nesfatin-1/NUCB2 in digestive system of the humans, Sprague-Dawley (SD) rats and the institute of cancer research (ICR) mice, so as to lay the foundation for further study of its functions in the digestive system. Methods The specimens were obtained from SD rats and ICR mice as well as 20 patients with digestive disease, who were admitted to the First hospital affiliated to Nanjing Medical University and receired surgercal treatment. The specimens from patients with malignant tumors were obtained 5 cm apart from cancerous tissues and from patients with benign tumors were obtained near the focus. The resected tissues included pancreas, stomach, duodenum, esophagus, liver, small intestine or colon. The distribution of nesfatin-1/NUCB2 was examined with immunohistochemical (IHC) staining and its protein level in each organ was measured using Western blotting. Results The immuinohistocemical study revealed the similar distribution pattern of nesfatin-1/NUCB2 in the digestive system of the patients, SD rats and ICR mice. Nesfatin-1/NUCB2 was found to localize in the center of the pancreatic islets, the lower 1/3 to 1/2 of the gastric mucosal glands, as well as the submucosa of the duodenum. Western blotting examination showed the expression of NUCB2 in all tissues from patients, SD rats and ICR mice, whereas the protein level of the nesfatin-1/NUCB2 was higher in pancreas (0.84±0.03, 0. 84±0.05 and 0. 84±0.04, respectively), stomach (0.86±0.06,0.81±0.02 and 0. 78±0.02, respectively) or duodenum (0.79±0.09,0. 79±0.04 and 0.78±0.05)than that in esophagus (0.43±0.04,0.44 ± 0.02 and 0.47 ± 0.06, respectively), liver (0.42±0.01,0.44±0.04 and 0.43 ± 0.01, objectively), small intestine (0.32±0.04,0. 32 ± 0. 04 and 0.34 ±0.04, respectively) or colon (0. 29±0.01,0.32±0.03 and 0. 28±0.03, respectively)(all P values=0. 000). Conclusion Nesfatin-1/NUCB2 is widely expressed in the pancreatic islets, gastric mucosal glands and duodenum of the patients, SD rats and ICR mice, which indicates that nesfatin-1/NUCB2 may be involved in the regulation of food intake, carbohydrate metabolism and gastrointestinal motility.

Objective To establish ulcerative colitis(UC)model in BALB/c mice and to investigate the role of high mobility group box chromosomal protein 1(HMGBI)in pathogenesis of UC.Methods Thirty-two BALB/c mice were randomly divided into UC group(n=24,which were fed with 3%dextran sulfate 80dium solution)and control group(n=8,which were fed with water).The animals were sacrificed at 24.96 and 1 68 hours,respectively,to collect samples of colon and blood.The sernm level of HMGB1 was measured with ELISA and the expression of HMGB1 in colon was determined by Western blotting analysis.Results Histological scoring increased with the induction of the model,and manifestation of colon mucosa at 168h was similar with that of UC in human.The serum level of HMGB1 was slightly higer at 24 h than that of control(5.09±0.61 μg/L vs 4.49±0.53μg/L,P＞0.05),and reached a peak at 96 h (14.74±0.60 μg/L,P＜0.01),decreased at 168 h(9.03±0.78μg/L,P＜0.01).The expression 0.05).significandy increased at 96h(0.76±0.03,P＜0.05)and at 168 h(0.77±0.04,P＜0.05).Conclusion HMGB1 might be involved in pathologic changes of UC at a later stage.

Objective To investigate cortactin expression in malignant colorectal tissues and corresponding adjacent non-cancerous colon tissues,precancerous lesions (adenomatous polyps) and the relationship between the expression of cortactin and cell division in colorectal cancer cells.Methods The expression of cortactin was detected by immunohistochemistry in colorectal cancer,colorectal adenomatous polyp (precancerous lesions) and colorectal tissues adjacent to adenocarcinomas (normal tissues).Kaplan-Meier method was employed to compare the survival between the groups.Cortactin expression and cell division were detected by Western blot and immunofluorescence in SW-620 colon cancer cells treated with cortactin siRNA.Results The positive expression rate of cortactin was significantly higher in colorectal cancer tissues than in adenomatous polyp tissues and pericarcinomatous normal tissues.Overexpression of cortactin in colorectal cancer tissues was correlated with poor differentiation (P ＜ 0.01),lymph node metastasis (P =0.006),and TNM stage (P =0.022).The 5 year survival rate of the group of negative/weak positive expression of eortactin was significantly higher than the group of strong positive expression of cortactin.CTTN gene amplification in colorectal cancer tissues was obvious.Cortactin siRNA induction caused a lower cortactin protein expression in colorectal cancer cells.Conclusions It is suggested that the excessive expression of cortactin contributes to the growth of cancer cells in colorectal cancer.

Objective:To observe the effect of Er Chen Tang on CYP2E1 and mitochondrial energy metabolism in nonalcoholic fat-ty liver disease ( NAFLD) to explore the role of Pinellinae Rhizoma Praeparata ( PRP) and Citri reticulatae pericarpium ( CRP) in the treatment of nonalcoholic fatty liver disease. Methods:Er ChenTang and the prescription without PRP or CRP was respectively given the animal models by gastric gavage. The serum levels of ALT, AST, triglyceride, cholesterol, SOD and MDA in hepatic tissue, and the contents of liver tissue CYP2E1 and ATP were detected in the mice. Results:The CYP2E1 levels in NAFLD mice increased signif-icantly with abnormal mitochondrial energy metabolism. Compared with those in the model group, the levels of ALT, AST, triglyceride and cholesterol were significantly reduced by Er Chen Tang, meanwhile, the content of CYP2E1 was reduced and also restored liver en-ergy metabolism. The treatment effect significantly decreased when the lack of PRP or CRP, and the ability of restoring liver mitochon-drial energy metabolism of Er Chen Tang decreased significantly when the lack of PRP (P<0. 05). After the removal of CRP, the in-hibition ability of Er Chen Tang to CYP2E1 levels significantly decreased (P<0. 05). Conclusion:Er Chen Tang can effectively im-prove nonalcoholic fatty liver diseases, and effectively reduce the content of CYP2E1 in liver tissue of mice and restore the mitochondri-al energy metabolism.

Objective To investigate the relationship between serum resistin,leptin and adiponectin with microangiopathy in the patients with type-2 diabetes mellitus(T2DM).Methods One hundred and twenty patients with T2DM in our hospital were selected and divided into the non-microangiopathy group (NON-MAP,60 cases) and microangiopathy group (MAP,60 cases) according to whether complicating microangiopathy.Other 60 individuals undergoing healthy physical examination were selected as the normal control group(NC).Fasting serum resistin,adiponectin and leptin levels were detected in each group.Fasting blood glucose,insulin and blood lipid levels were also detected.The insulin resistance was evaluated by using insulin resistance index(HOMA-IR).Results The levels of serum resistin,leptin,free fat acid(FFA) and hs-CRP in the NON-MVP group and MVP group were signifi cantly higher than those in the NC group,while the adiponectin level was sigrnificantly lower than that in the NC group,the difference was statistically significant(P＜0.05).The correlation analysis showed that serum resistin and leptin levels had positive correlation with hs-CRP,FFA,HOMA-IR and TG(P＜0.05),and had negative correlation with HDL-C(P＜0.05);the adiponectin level was negatively correlated with hs-CRP,FFA,HOMA-IR and TG,while positively correlated with HDL-C(P＜0.05);serum resistin and leptin levels had positive correlation,both had significantly negative correlation with adiponectin;with serum resistin,leptin and adiponectin as the dependent variables,the multiple stepwise linear regression analysis showed that HOMA-IR and waist to hip ratio had maximal influence on them.Conclusion Serum resistin and leptin levels increase and adiponectin level decrease in T2DM patients suggests that serum resistin,leptin and adiponectin are correlated with T2DM occurrence as well as microangiopathy occurrence.

Objective: To investigate the protective effects of matrine combined with glycyrrhizic acid on chronic liver injury induced by carbon tetrachloride, and explore the protective mechanism from the points of energy metabolism and CYP enzyme.Methods: The chronic hepatic injury model of rats was induced by CCl4.The changes of activity of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in serum were measured to observe the protective effect of the two drugs and their combination.The contents of glutamate dehydrogenase (GLDH) in serum and adenine nucleoside three phosphate (ATP), adenosine diphosphate (ADP) and adenine monophosphate (AMP) in liver tissue were determined to evaluate the regulation effect on hepatic energy metabolism and mitochondrial function.The levels of CYP1A2, CYP2E1 mRNA and protein in liver tissue were detected by real-time PCR and Western Blot to evaluate the two drugs and their combination on the regulation function of liver CYP enzyme.Results: Matrine (72.8 mg×kg-1)and glycyrrhizic acid(43.4 mg·kg-1)could decrease the serum activities of ALT and ALT in chronic hepatic injury model, and the combination (matrine 36.4 mg·kg-1+glycyrrhizic acid 21.7 mg·kg-1) had the most significant protective effect (P＜0.05).Matrine (72.8 mg·kg-1)and glycyrrhizic acid(43.4 mg·kg-1)could decrease GLDH in serum,and restore the content of ATP in liver (P＜0.05).Matrine (72.8 mg·kg-1) had no effect on the expression of CYP1A2 and CYP2E1mRNA, and glycyrrhizin (43.4 mg·kg-1) could inhibit the expression of CYP1A2, CYP2E1mRNA and protein (P＜0.05).Conclusion: Matrine combined with glycyrrhizin has obvious regulation effect on mitochondrial function and liver protective effect in chronic hepatic injury model.

Objective To investigate the effect of different frequencies of endoscopic ultrasonography (EUS) on diagnosis of duodenal submucesal lesions. Methods A total of 210 elevated duodenal lesions detected by routine endoscopy in 201 patients were examined by EUS at different frequencies according to lesion features and image clarity. Endoscopic resection was applied in 27 lesions with informed consents,and pathological findings were compared with the results of EUS. Results The accuracy of EUS was 81.4% (22/27). Ultrasound of low frequency was superior in displaying overview of lesion and was more suitable for lesions larger than 1.5 cm in diameter or lesions of hyperecho. On the other hand,ultrasound of high frequency was more reliable to reveal minute structures and could be employed in scanning lesions smaller than 1.5 cm in diameter. Combination of different frequencies of ultrasound could provide more details in larger lesions. Conclusion Application of appropriate frequency or combined frequencies of ultrasound in examination of elevated duodenal lesions is beneficial to achieve more accurate diagnosis.

Combining with the data based on data mining,to collect the clinical experimental research ralating to thermal moxibustion from June 2010 to March 2015, and extract the representative clinical experimental research of clinical diseases. To summarize advances in clinic study of Thermal Moxibustion in recent five years.