background The lead isotope ratios (LIR) differ among different sourced samples. Previous domestic and oversea studies on source tracing by LIR in human blood or urine mainly focused on the comparison of blood or urine samples from the same or different individuals, while few comparisons between biological and environmental samples, and the reported relative standard deviations (RSDs) of the main LIR (^207/206Pb and ^208/206Pb) fluctuate widely from 0.3% to 1%. Objective To optimize inductively coupled plasma mass spectrometry (ICP-MS), obtain a better RSD, and determine LIRs of human blood, urine, and related environmental samples. Methods The ICP-MS was optimized for operating conditions and parameters according to the sensitivity and RSD of LIR. The study subjects were 40 lead-exposed workers in a lead-acid battery factory and 2 lead poisoned children in a hospital. The samples included 40 blood and 40 urine samples from the workers before shift, 4 dust samples and 2 water samples in the workplace on the same day before shift, 2 blood and 3 urine samples from the children before hospital admission due to lead-poisoning, and 4 urine samples after medical treatment. After heating and acid digestion, the LIR (^207/206Pb and ^208/206Pb) of biological and environmental samples were determined by the optimized ICP-MS method. t-test and two-dimensional traceability graphics were adopted to analyze the detection results. Results The calibrated RSDs of the LIR (^207/206Pb and ^208/206Pb) of lead isotope standard solution were 0.11% and 0.08% respectively, and the NIST-SRM-981 actual values were 0.91531±0.00097 and 2.1670±0.0017, respectively. When the total concentration of lead was greater than 5 μg·L⁻¹, the RSD of each isotope ratio was stable gradually; when the total concentration of lead was between 10-80 μg·L⁻¹, the RSD was below 0.20%. There were statistically significant differences in the blood and urine LIR (^207/206Pb and ^208/206Pb) of the lead-exposed workers (t=5.831, P<0.001; t=21.021, P<0.001), the LIR (^207/206Pb and ^208/206Pb) between workplace dust samples and workers’ urine samples (t=−6.879, P=0.038; t=12.521, P<0.001), and the ^208/206Pb between workplace dust samples and workers’ blood samples (t=−10.46, P<0.001), except the ^207/206Pb between workplace dust samples and workers’ blood samples (t=−0.12, P=0.912). In the patients afflicted with lead poisoning, the projection points of LIR of blood and urine samples from the same individual were not at the same level in the two-dimensional model, nor was the LIR of urine samples before and after medical treatment of the same individual. Conclusion The optimized ICP-MS can control the RSD of main LIR (^207/206Pb and ^208/206Pb) below 0.20%. There are differences in the LIR distributions of different samples.

Objective To investigate the mechanism of occurrence and development of zinc-alpha-2-glycoprotein (AZGP1) in the activated hepatic stellate cells (HSCs) and liver fibrosis.Methods The activated human hepatic stellate cell line LX2 was induced by the stimulation of transforming growth factor-β1 to construct carbon tetrachloride liver fibrosis mice model.The situation expression of AZGP1 in liver cells and tissues were observed.Plasmid transfection method was used to detect the activation,proliferation,apoptotic functions and changes in related factors of LX2 cells,respectively,after the overexpression and inhibition of AZGP1expression.Univariate analysis of variance was used for multiple group comparison.Results The results of immunofluorescence staining showed that AZGP1 protein was decreased and α-smooth muscle actin was increased in the activated LX2 cells,and the two were negatively correlated.AZGP1 gene and protein were significantly under-expressed in activated LX2 cells and liver tissues of mice with carbon tetrachloride liver fibrosis.Collagen I,matrix metalloproteinase-2,and α-smooth muscle actin genes and proteins were significantly down-regulated in LX2 cells after over-expression of AZGP 1.Cell fluorescence showed that AZGP 1-overexpressing cells were activated and α-smooth muscle actin protein was reduced.In addition,the proliferative activity and G1/S-specific cyclin D1 protein of LX2 cells were significantly reduced after overexpression of AZGP1,while cell cycle experiments showed that the proportion of cells overexpressing AZGP1 was significantly increased in the G0/G1 phase,and the proportion of S phase was significantly reduced.AZGP1 had no significant effect on the apoptosis of LX2 cells.Conclusion AZGP1 can reverse liver fibrosis by inhibiting the activation and proliferation of hepatic stellate cells,and thereby overexpression of AZGP1 is expected to become a new target for liver fibrosis treatment.

Objective: To discuss the affect of glycosylated hemoglobin (HbA1c) level for the onset of nonalcoholic fatty liver disease (NAFLD) in cohort population. Methods: An epidemiological survey of the relationship between HbA1c and NAFLD conducted in 2012 was based at cohort baseline, and three follow-up sessions conducted in 2013, 2014 and 2015. In total 2 811 subjects were included in the study after exclusion of NAFLD patients at baseline and those who lost their lives due to relocation, and death. The Cox proportional hazard model was used to analyze the relationship between glycosylated hemoglobin and other risk factors of NAFLD. Continuous variables were compared using the t-test or the Mann-Whitney test. χ ²-test was used for the measurement of categorical data. Results: A total of 2 811 subjects with mean age of 59 (58.2±9.8) years old, including 1 664 males and 1 147 females. Age, waist circumference, body mass index, systolic blood pressure, γ-glutamyltransferase and fasting blood glucose level of HbA1c abnormal group were higher than normal group. The incidence of NAFLD in the abnormal HbA1c level group (25.4%) was higher than normal group (14.9 %), and diastolic blood pressure, high-density lipoprotein cholesterol was lower than normal group and the differences were statistically significant. During the three follow-up intervals, there were 440 new cases of NAFLD, consisting 285 males and 155 females with cumulative incidence of 15.7% (440/2 811). Multivariate Cox regression analysis showed that patients with elevated HbA1c had a higher risk of developing NAFLD (HR 1.796; 95% CI 1.335~2.418; P < 0.01), and the increased HbA1c level after adjustment for gender, age, and metabolic syndrome-related factors remained an independent risk factors for NAFLD (HR 1.580; 95.0% CI 1.161-2.152; P < 0.01). Conclusion An elevated HbA1c levels have a positive predictive value for the onset of NAFLD.

Objective To study the correlation between non-alcoholic fatty liver disease (NAFLD)and glycosylated hemoglobin (HbA1 c) in middle-aged and aged population.Methods A total of 4 127 inservice workers and retirees aged 45 years old or above from one petrochemical enterprise in Ningbo were enrolled in our study.The waistline,body mass index,blood pressure,fasting blood-glucose,blood lipid profile,glutamyltranspeptidase,HbA1c and epigastrium B ultrasound were investigated.According to the quartile of HbA1c level,participants were divided into four groups,namely,Q1 group ≤5.2％,Q2 group ＞ 5.2％-5.4％,Q3 ＞ 5.4％-5.6％ and Q4 group ＞ 5.6％.The prevalence of NAFLD and clinical characteristics of each group were analyzed.Logistic regression was used to predict independent risk factors of NAFLD.Results The morbidity of NAFLD was 27.2％ with 31.9％ in male and 21％ in female,which was significantly higher in men.In Q1,Q2,Q3 and Q4 group,the prevalence of NAFLD were 18.5％ (178/961),22.8％ (185/812),25.6％ (280/1 095),38.1％ (480/1 259) respectively.With the increase of HbA1 c level,the morbidity of NAFLD increased synchronously.The age,systolic pressure,total cholesterol,low densitylipoprotein cholesterin and fasting blood-glucose were all elevated according to the increase of HbA1 c in 1 123 NAFLD patients.Multi-factor logistic regression analysis indicated that high HbAlc level was the risk factor of NAFLD (OR =1.67,95％ CI 1.15-2.43,P =0.007).Conclusion HbA1c is an independent risk factor of NAFLD and both of these are closely related to blood lipid metabolism disorder.

Purpose To investigate relationship between PD-L1 molecule expression and Treg infiltration in non-small cell lung cancer ( NSCLC) tissue and to explore their clinical significance. Methods Immunohistochemistry was used to detect the PD-L1 molecules expression and Treg infiltration of 78 NSCLC tissues. The relationship among PD-L1 expression, Treg infiltration and clinic-pathological parameters was analyzed in the patients. Results PD-L1 molecule was highly expressed in lung cancer tissues than adjacent tissues (52. 5% vs 6. 4%), and same to Foxp3 +Treg (18. 63 ± 16. 67)/HPF vs (2. 96 ± 2. 97)/HPF. There was close relationship between PD-L1 expression and Treg infiltration with lymph node metastasis and clinical stage of patients, but was no statistical correlation with patient’ s age, gender, histological type and degree of differentiation of the tumor cells. PD-L1 expression was significantly higher in advanced stage than that in early stage (70. 0% vs 41. 7%) and same to Treg infiltration (73. 3% vs 35. 4%). There were also signif-icantly higher infiltration with lymph node metastasis than that without metastasis. In addition, PD-L1 molecule expression and Foxp3 +Treg infiltration were positively correlated (rs =0. 611, F=78. 82, P=0. 023). Conclusion There was strong relationship among PD-L1 expression, Treg infiltration and disease progress in lung cancer patients, and they possibly participate in the progression and immune escape of lung cancer.

Objective To compare the value of bedside index for severity in acute pancreatitis (BISAP),Ranson score and Balthazar computed tomography severity index (CTSI) in predicting the severity and prognosis of acute pancreatitis (AP).Methods From 2005 to 2011 in Shanghai,the clinical data of 1004 AP cases from seven hospitals was collected and retrospectively analyzed.The value of BISAP score,Ranson score and Balthazar CTSI in predicting the severity and prognosis of AP were assessed with receiver operator characteristic (ROC) curve.Results Among 1004 patients,the main cause of AP was biliary disease (580 cases),about 57.77％.The incidence of pancreatic necrosis,mortality and SAP increased along with BISAP score.The risk of pancreatic necrosis in patients with CTSI ≥ three was significantly higher than that of ＜ three.The risk of pancreatic necrosis and SAP in patients with BISAP score ≥ two was significantly higher than that of ＜ two (OR:4.93,95％CI 3.62-6.70; OR 2.62,95％CI 1.59-4.31,respectively).There was no significant difference in the accuracy of predicting the progression and mortality of AP among these three score systems.However the sensitivity of BISAP score (OR:61.54,95％CI 35.09-87.99) in predicting the progression and mortality of AP was better than that of Ranson (OR:46.15,95 ％ CI 19.05-73.25) and CTSI (OR:46.15,95％CI 19.05-73.25).Conclusions BISAP score is easy to perform and when combined with CTSI,it helps to make the diagnosis and classification of AP in time,predict the prognosis accurately.Compared with Ranson score,BISAP score has higher clinical value.

Objective To discuss the surgical procedures and treatment after combined liver and intestinal transplantation with portal venous drainage and enterostomy of two ends in one case.Methods A male patient with liver dysfunction and short bowel syndrome underwent the combined liver and intestinal transplantation.With the techniques of en bloc,the liver and intestinal grafts were harvested from cadaveric donor.The intestinal graft,200 cm long,was implanted with portal venous drainage and aortic inflow,and enterostomy of both ends was performed instead of intestinal anastomosis.The liver graft was placed in a piggyback fashion with end to end anastomosis of the bile ducts without T tube. Inmunosuppression protocol was administrated with campath-1H and tacrolimus.Endoscopic biopsy of intestinal graft was performed regularly,and clinical observation was done to monitor the acute rejection.Results In the first month after operation,abdominal infection was controlled by intraperitoneal drainage with open surgery.One suspect acute rejection was treated with methylprednisolone.Until sixth month,the functions of liver and intestine were progressively restored.However,the patient lost weight and could not be free from intravenous nutrition because of diarrhea.Conclusion Combined liver and intestinal transplantation with portal venous drainage and enterostomy of two ends is a simple surgical procedure with lower risk of surgical complications.This method is propitious to monitoring rejection and function improvement of the grafts.Diarrhea and loss of digestive juice are the main reasons of low body weight and malnutrition.

Objective To evaluate the diagnostic value of intraductal ulstrasonography(IDUS)in non-opaque bile duct stones.Methods Between January 2009 and August 2010 in the Department of Gastroenterology at Shanghai 10th People's Hospital,a total of 183 patients(male:102 cases,mean age 69 years； female:81 cases,mean age 71 years)were enrolled,who were suspected of bile duct stones or stenosis which could not be diagnosed by abdominal CT,MRI,and abdominal B-mode ultrasound.All the patients underwent endoscopic retrograde cholangiopancreatography(ERCP)first,and then patients with non-opaque bile duct stones followed by IDUS.Results A total of 134 cases (73.2％)of bile duct stones were diagnosed by ERCP,49 cases(26.8％)were negative.And then the 49 patients underwent IDUS,of whom 24 patients with sand-like stones,11 patients with lowdensity stones,6 patients with ampullary cancer,2 patients with pancreatic cancer,6 patients with sclerosing cholangitis.The diagnostic accuracy of IDUS in the position and quality of bile duct stones was 100％,higher than that of ERCP,which was 80％.After ERCP,pancreatitis occurred in 3 patients and improved after conservative treatment,there was no complications like perforation and bleeding.Conclusion The diagnostic accuracy of IDUS in the position and quality of bile duct stones is high,which can make up for the misdiagnosis by ERCP without increasing the complications.IDUS can provide reliable basis for the diagnosis of clinical bile duct stones.

Objective To investigate the role of pancreatic β cell on pancreatic regeneration following experimental acute pancreatitis.Methods Eighty-seven SD male rats were randomly divided into four groups:control group ( n =15 ),STZ group ( n =24),L-Arg group ( n =24 ),STZ + Arg group ( n =24).60 mg/kg of STZ was administrated by intraperitoneal injection to induce the diabetes model.2.5 g/kg body weight of LArg was administrated by intraperitoneal injection to induce the acute pancreatitis model.The rats were sacrificed 1,3,5,7 d later and the serum levels of amylase and glucose were measured.Relative pancreatic weight (pancreatic weight/body weight) were measured.Pancreatic tissue underwent routine pathologic examination,and the percentage of area of necrosis and tissue transformation was calculated.The expression of Reg4 and insulin was performed by immunofluorescence.Results Serum level of glucose significantly increased after STZ injection.After L-Arg injection,serum level of amylase significantly increased,and there was pancreatic tissue edema,necrosis,infiltration of inflammatory cells,which suggested the successful model induction.The percentage of area of necrosis in STZ + L-Arg group was (71.6 ± 6.0) ％ at the 3rd day,which were significantly higher than (42.3 ± 4.0 ) ％ in L-Arg group; the percentage of area of transformation was (45.6 ± 5.4) ％,which were significantly lower than (78.5 ± 6.4) ％ in L-Arg group.Expression of Reg4 in pancreatic islets of STZ + L-Arg group was significantly lower than those in L-Arg group.Conclusions STZ impairs pancreatic β cells,aggravates pancreatic damage following L-arginine induced pancreatitis and inhibits pancreatic regeneration.

Objective To investigate the expression of Iroquois homeobox protein 1 ( IRX1 ) gene and the hypermethylation status of its promoter in pancreatic cancer,and their relationship.Methods Real-time PCR was used to quantitatively detect IRX1 gene expression level of 12 sets of resected pancreatic cancer tissue and 6 sets of pancreatic cancer cell lines; gene sequences analysis was used to detect the structure of IRX1 promoter; DNA methylation inhibitor 5-Aza-2′-deoxycytidine (5-Aza-dC) was used in pancreatic cancer cell lines,and then the methylation of IRX 1 was measured by methylation-specific PCR (MSP) and unmethylation sequence-PCR (USP) methods.Results Expression of IRX1 mRNA in pancreatic cancer tissue was 0.31 ± 0.11,which were significantly lower than that in normal pancreatic tissue ( 1.05 ±0.32,P ＜0.01 ).IRX1 mRNA expression of AsPCl,BxPC3,Capan 2,PANCl,PaTu8988 and SW1990 were 0.36 ± 0.08,0.34 ±0.16,0.37 ±0.11,0.25 ±0.06,0.31 ±0.04,0.36 ±0.02,which were significantly lower than that in human kidney epithelial 293 cells ( 1.03 ± 0.28,P ＜ 0.05 or ＜ 0.01 ).Analysis of IRX1 gene sequence showed abundant CpG islands in promoter.Hypermethylation of IRX1 promoter site was found in all pancreatic cancer cell lines.However,its methylation status could be reversed by 5-Aza-dC,and the IRX1 expression was also restored.Conclusions IRX1 mRNA expression is down-regulated in pancreatic cancer,and it is related with promoter CpG islands hypermethylation.