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 investigate the expression of immunoglobulin-like transcript 3 (ILT3) in colorectal cancer tissues and colorectal cancer cell lines,and the correlations with the clinicopathological factors and prognosis,and to reveal the clinical significance of ILT3 abnormal expression in colorectal cancer.Methods The mRNA and protein expressions of ILT3 in colorectal cancer cell lines SW480,SW620,HT-29 and human normal intestinal epithelial cell line FHC were detected by real-time fluorescence quantitative PCR and Western blotting,and the differences were analyzed.Immunohistochemistry was used to detect the expressions of ILT3 in 85 colorectal cancer tissues and 30 adjacent normal tissues.The correlations between the expression of ILT3 and clinicopathological factors and prognosis were analyzed.Results The relative expressions of ILT3 mRNA in cancer cell lines SW480,SW620,HT-29 and the normal intestinal epithelial cells line FHC were 10.813 ± 3.289,4.391 ± 0.622,1.481 ± 0.055 and 1.000 ±0.000 respectively,with a significant difference (F =21.846,P ＜ 0.001).Compared with the normal intestinal epithelial cell line FHC,the relative expressions of ILT3 mRNA in cancer cell lines SW480 and SW620 were significantly higher,with a statistically significant difference (P ＜ 0.001,P =0.038),and the relative expression of ILT3 mRNA in cancer cell HT-29 was higher,but the difference has no statistical significance (P =0.734).The relative expressions of ILT3 protein in cancer cell lines SW480,SW620,HT-29 and normal intestinal epithelial cells line FHC were 1.838 ±-0.651,1.763 ± 0.222,1.449 ± 0.200 and 0.628 ± 0.263 respectively,with a statistically significant difference (F =6.320,P =0.017).Compared with the normal intestinal epithelial cell line FHC,the relative expressions of ILT3 protein in cancer cell lines SW480,SW620,HT-29 were significantly higher,with a statistically significant difference (P =0.005,P =0.007,P =0.030).ILT3 was mainly expressed in the membrane and (or) cytoplasm.In 85 cases of colorectal cancer tissues,55 cases (64.7％) have ILT3 high expressions,which were significantly higher than those in the adjacent normal tissues (16.7％,5/30),with a significant difference (x2=20.508,P ＜ 0.001).The high expression of ILT3 was related tolymph node metastasis (x2 =4.684,P =0.030) and TNM staging (x2 =4.684,P =0.030),but it was not related to age (x2 =0.927,P =0.336),gender (x2 =0.078,P =0.780),tumor location (x2 =0.249,P =0.618),tumor size (x2 =1.813,P =0.178) and tumor differentiation (x2 =0.807,P =0.369).The median overall survival of patients with high ILT3 expression was significantly shorter than that of patients with low ILT3 expression (45.0 months vs.74.0 months),with a significant difference (x2 =17.907,P ＜ 0.001).Conclusion The expressions of ILT3 in colorectal cancer cells are higher than those in the normal intestinal epithelial cell line,the expressions of ILT3 in colorectal tissues are higher than those in the adjacent normal tissues,which are correlated with lymph node metastasis and TNM staging.Patients with high expressions of ILT3 indicate poor clinical prognosis.The expression of ILT3 may serves as a new target for diagnosis,treatment and prognosis of colorectal cancer.

Sorafenib is currently the only systematic molecular targeted drug approved by many coun-tries for the treatment of advanced HCC.But sorafenib limites has survival improvement,patients appear inevi-table disease progression soon.In recent years,Oxaliplatin-based regimens showed promising anti-tumor activi-ty,safety and low toxicities,which provides a new way for treatment of advanced HCC.

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 analyze expression of interleukin (IL)-23p19 and IL-23 receptor (IL-23R) in inflammatory bowel disease (IBD),and the role in the induction of peripheral blood T cell activation and proinflammatory cytokine secretion.Methods Peripheral blood (PB) and intestinal mueosal biopsies were collected from 12 patients with Crohn's disease (CD),25 patients with ulcerative colitis (UC) and 20 healthy controls.Expression of IL-23p19 was determined by immunohistochemistry and RT-PCR.IL-23R expression in CD4+,CD8+ T and NK cells from peripheral blood and lamina propria was analyzed by flow eytometry.Peripheral blood mononuclear ceils (PBMC) were isolated and cultured under stimulation with IL-23 and anti-CD3,and the levels of tumor necrosis factor α (TNFα) interferon (IFN)γ and IL-2 were determined by enzyme-linked immunosorbent assay (ELISA).Results The expression of II.-23p19 was significantly increased in inflamed mucosa of CD at both the transcriptional and translational levels compared with that in UC and healthy controls.IL-23R was mainly expressed in PB- and lamina propria-CD4+,CD8+T cells and NK cells from IBD patients,and markedly increased compared with controls (P＜0.05).IL-23 strongly triggered PBMC from IBD patients to produce significantly higher levels of IFN-γ,TNF-α and IL-2(P＜0.05).Conclusions IL-23p19 and IL-23R are highly expressed in IBD,particularly in CD,and may play an important role in the induction of T cell activation and proinflammatory cytokine secretion,suggesting that targeted therapy directed against IL-23 may have a therapeutic role in IBD.

Objective To investigate the role of Hedgehog pathway in hepatic fibrosis and its association with activation of hepatic stellate cells. Methods Twenty male Spragur-Dawley rats were divided into control and model groups with 10 each. The animal models were induced by injection with CCl4 and fed with fat-rich diet. The rats in both groups were sacrified at the 8 week with 5 each and the liver tissues were removed for HSC-T6 culture. The deposition of collagen fiber in liver was detected with HE and Masson staining. RT-PCR was used to detect the expressions of Sonic hedgehog (Shh), smoothened (Smo), patched (Ptc), Gli-1 and α-smooth muscle action (α-SMA) mRNA in HSC-T6 and liver tissues. The influence of cyclopamine (Cyc) and lipopolysaccharide (LPS) on HSC-T6 proliferation were assayed by MTT. The expressions of Shh, Smo, Ptc, Gli-1 and α-SMA mRNA after intervention with Cyc (100μmol/L) and LPS were measured by real-time PCR. Results A lot of lipo and collagen deposited in liver of model rats. The Shh,Smo,Gli-1 and α-SMA mRNA were highly expressed in model rats than those in control group (2-△△Ct were 20.45±3.31 vs. 1, 12.78 ± 0. 53 vs. 1, 10.88 ± 2.41 vs. 1, 4.91 ± 2. 59 vs. 1, respectively, all P value <0. 05). In vitro Cyc inhibited HSC-T6 proliferation in dose dependant manner (F=636.81, P<0.01). Compared to the control group, the mRNA expressions of Smo, Ptc, Gli-1,α-SMA in HSC-T6 were significantly reduced after Cyc intervention (2△△Ct, were 0. 20±0. 11, 0. 21 ± 0. 08, 0. 28 ± 0. 05,0. 27±0.10,respectively, all P values<0.01). Conclusion The expression of members of Hedgehog pathway are increased in the progress of hepatic fibrosis, which may accelerate the hepatiee fibrosis by activating HSC.

Objeetive To observe the curative effect of ultramicro-dosage heparin (UMDH) combining with low-molecule dextran (LMD) on the D-dimer positive infant with severe pneumonia.Methods sixty-eight infant patienls.whose D-dimer in serum were positive,were randomly divided into two gmaps cutaneous injection for 4 times.Arier the D-dimer in serum became negative,the drug was stpped.In the heparin combining with LMD (HCLMD) group,the UMDH (same dosage as above) was used combining with the LMD.The dosage for LMD was 5 ml/kg once with general-speed intravenous drip and 1-2 times a day,which should be continued until the D-dimer in serum became negative.Results After 2-day's trestment,there was no obvious differences in the negative transfer rate of D-dimer between two groups(P＞0.05).However,after 3-4 day's treatment,the negative transfer rate in HCLMD group was absloutely higher than that in heparin group,there were obvious differences in two groups(P＜0.05).As for the death rate of illness,the death rate in HCLMD group was lower than that in hepari.group,being respectively 5.88% and 17.65%.Conclusion The use of UMDH combining with small-dosage LMD dextran can quickly improve the high condemation situation of blood and reverse the condition of infant patients with severe pneumonia.

Respiration is one of the most important modulators of heart rate variability(HRV). The modulation mechanism of respiration on HRV were introduced. The effects of respiratory rate, respiratory volume and respiratory patterns on HRV were discussed, and these effects on HRV in patients were also referred. These respiratory effects should be taken into account when HRV is used to predict cardiac vulnerability.

Objective To investigate the effects and mechanism of Safflower Injection on the proliferation,apoptosis,and expression of bcl-2/bax gene in hepatic stellate cell(HSC) in vitro.Methods HSC Line HSC-T6 was incubated with Safflower Injection at different concertration,cell proliferation was assessed by MTT colorimetric assay.After incubated with Safflower Injection 5,10,and 20 mg/mL,flow cytometry(FCM) was used to detect the content of DNA in HSC-T6.Morphological change of HSC-T6 was observed under microscope and agarose gel electrophoresis for DNA Ladder was used to detect apoptosis.Besides,the early stage of apoptosis was detected with Annexin V-FITC/PI double labbled assay.And real time RT-PCR was used to detect the expression of bcl-2/bax gene.Results The significant inhibition of Safflower Injection on HSC proliferation was observed in a dose-and time-dependent manner.Observed by FCM,the cell ratio in G0/G1 phase with Safflower Injection treatment(10 and 20 mg/mL) for 24 h was increased,which showed the significant difference compared with the control group(P