Objective To investigate the early diagnosis and disease evaluation value in patients with acute pancreatitis by various serum cytokines measurement.Methods Forty-eight acute pancreatitis patients were divided into two groups based on the results of computed tomography (CT) examination:mild acute pancreatitis group (30 cases) and severe acute pancreatitis group (18 cases).The other 30 normal persons were selected as control group.The various serum cytokines were measured by enzyme-linked immunosorbent assay (ELISA).Results The serum concentrations of interleukin(IL)-1,IL-10 and tumor necrosis factor (TNF)-α in mild acute pancreatitis group were significantly higher than those in control group [(25.00 ± 1.92) ng/L vs.(10.08 ± 2.65) ng/L,(59.78 ± 4.51) ng/L vs.(1.80 ± 0.66) ng/L,(55.31 ± 8.54) ng/L vs.(18.72 ± 7.84) ng/L,P ＜ 0.05].The serum concentrations of IL-1,IL-6,IL-8,TNF-α and platelet activating factor (PAF) in severe acute pancreatitis group were significantly higher than those in mild acute pancreatitis group [(93.27 ± 3.98) ng/L vs.(25.00 ± 1.92) ng/L,(397.84 ± 13.05) ng/L vs.(34.12 ± 4.96) ng/L,(93.32 ±3.40) ng/Lvs.(13.06± 1.86) ng/L,(181.94 ±7.54) ng/Lvs.(55.31 ±8.54) ng/L,(284.53 ±7.88) ng/L vs.(175.25 ±30.15) ng/L,P＜0.05].Conclusion The various serum cytokines measurement has great importance on the early diagnosis of acute pancreatitis and discrimination between the mild acute pancreatitis and severe acute pancreatitis.

Objective To explore the mutation situation of pancreatic cancer cell mitochondria DNA D-loop region. Method PCR and direct sequencing was used to analyze the mutational site of mitochondria DNA D-loop region in two pancreatic cancer cell lines SW1990 and JF-305 and normal primary cultured pancreas cell. Result The point mutations were found in two pancreatic cancer cell lines and normal primary cultured pancreas cell. Eight point mutations were found in SW1990 and 9 point mutations were found in JF-305. Three point mutations (73 site A-G,16223 site C-T, and 16358 site c-T) existed in all two pancreatic cancer cell lines and normal primary cultured pancreas cells, which can be considered as polymorphism. Other two point mutations (16211 site C-T and 16311 site T-C) were only found in two pancreatic cancer cell lines, which can be considered as special mutations. Conclusion The mitochondrial DNA D-loop region of pancreatic cancer cells existed polymorphism and special mutations, and the special mutations might be new molecule marker.

Objective To explore the association of genetic polymorphism of fucosyltransferase (FUT) 2 and FUT3 and expression of Lewis antigen with ulcerative colitis (UC) in Chinese Zhejiang Han population.Methods We genotyped FUT2 (rs281377, rs1047781 and rs601338) and FUT3 (rs28362459, rs3745635 and rs3894326) in 485 UC patients and 580 healthy controls using SNaPshot. By immunohistochemistry method, we also evaluated expression of Lewis a and b antigens in the sigmoid colon of 10 UC patients and 10 patients with benign colonic polyps.Results The frequencies of mutant allele (A) and genotype (GA+AA) in FUT3 rs3745635 were higher in UC patients than in controls (P=0.016, 95%CI 1.339-1.699;P=0.038, 95%CI 1.330-1.742, respectively). Stratified analyses revealed that the frequencies of mutant allele (G) and genotype (TG+GG) of FUT3 rs28362459 were significantly lower in patients with extensive colitis than in those with distal colitis (P=0.001, 95%CI 0.567-0.786;P<0.001, 95%CI 0.503-0.742, respectively). Similar conclusions were drawn for the mutant allele (A) and genotype (GA+AA) of FUT3 rs3745635 in patients with extensive colitis compared to those with distal colitis (P=0.011, 95%CI 0.621-0.900;P=0.006, 95%CI 0.553-0.845, respectively). Although expression of Lewis b antigen in the sigmoid colon did not differ between UC patients and controls, Lewis a antigen expression was higher in the crypt epithelium of both inflammatory and non-inflammatory sigmoid colon of UC patients than in controls (P=0.028).Conclusion Polymorphisms of FUT3 and expression level of Lewis a antigen might be associated with UC.

OBJECTIVETo assess the association of transcobalamine II (TCN2) gene polymorphisms and serum levels of homocysteine (Hcy), vitamin Band folate with ulcerative colitis (UC) among Chinese patients.

METHODSFor 397 UC patients and 574 controls, two single nucleotide polymorphisms of the TCN2 gene (rs1801198, rs9606756) were tested with an improved multiple ligase detection reaction method. Serum Hcy, vitamin Band folate were measured with an enzymatic cycling assay and an chemiluminescence immunoassay, respectively.

RESULTSThe allelic and genotypic frequencies of rs1801198 and rs9606756 did not differ significantly between the two groups (all P> 0.05). Compared with those of the control group, the frequencies of G allele and CG+GG genotype of rs1801198 were greater in patients with moderate and severe UC (both P< 0.05). The same conclusion may also be drawn for the G allele and AG genotype of rs9606756 (both P< 0.05). Compared with the controls, average Hcy level was enhanced in UC patients (P< 0.01), whereas average vitamin Band folate levels were decreased in UC patients (both P< 0.01). In both groups, the average level of Hcy was lower in individuals carrying CC of (rs1801198) than in those with CG+GG (both P< 0.05). A similar conclusion was also drawn for individuals with AA of rs9606756 when compared with those carrying AG(both P< 0.05). Compared with patients with mild UC, average Hcy level was increased in those with moderate and severe UC (P< 0.01), while average vitamin Band folate levels were decreased in those with moderate and severe UC (both P< 0.01). The prevalence of hyperhomocysteinemia(HHcy), vitamin Bdeficiency and folate deficiency was greater in UC patients than in controls (all P< 0.01). In UC patients, the level of Hcy was negatively correlated with those of vitamin B(P< 0.01), albumin(P< 0.01), red blood cells(P< 0.01) and platelet (P< 0.05), but positively correlated with white blood cells(P< 0.01) and Mayo score (P< 0.01). Both HHcy and folate deficiency were independent risk factors for UC (OR=4.173, OR=5.206, both P< 0.01).

CONCLUSIONTCN2 (rs1801198, rs9606756) variations, as well as serum levels of Hcy, vitamin Band folate, are correlated with UC. Both HHcy and folate deficiency are independent risk factors for UC.

Adult ; Colitis, Ulcerative ; blood ; etiology ; genetics ; Female ; Folic Acid ; blood ; Genotype ; Homocysteine ; blood ; Humans ; Male ; Middle Aged ; Polymorphism, Single Nucleotide ; Transcobalamins ; genetics ; Vitamin B 12 ; blood

Objective: To investigate the influence of tumor necrosis factor-related apoptosis-inducing ligant (TRAIL) deficiency on mice colitis and the gut microbiota composition by inclding the expermental colitis model in tumor necrosis factor-related apoptosis-inducing ligand gene knockout (TRAIL^-/-) mice. Methods: C57BL/6 TRAIL^-/- mice and wild type (WT) mice were selected and assigned into TRAIL^-/- control group (eight mice), TRAIL^-/- colitis group (16 mice), WT control group (eight mice) and WT colitis group (16 mice). The mice of two colitis groups were oral administrated with 3.5% dextran sulphate sodium (DSS) in drinking water for seven consecutive days to induce experimental colitis model. The severity of colitis was evaluated by clinical appearance and histopathological examination. The colonic tissue samples of mice were collected and microbiota profile was analyzed by 16S rDNA sequencing method. USEARCH software and R language were used to analyze the difference of gut microbiota among TRAIL^-/- control group, TRAIL^-/- colitis group, WT control group and WT colitis group. T test and Mann-Whitney U test were used for statistical analysis. Results: After modeling, the disease activity index (DAI) of WT colitis mice and TRAIL^-/- colitis mice both gradually increased over time. Furthermore, compared with colitis mice, TRAIL^-/- colitis mice developed body weight loss, diarrhea and hemafecia earlier. On the seventh day after modeling, the percentage of body weight loss of TRAIL^-/- colitis mice and WT colitis mice was (28.98±2.84)% and (17.87±3.70)%, respectively; and the difference was statistically significant (t=9.53, P<0.01). The length of colon of TRAIL^-/- colitis mice was shorter than that of WT colitis mice ((4.63±0.28) cm vs. (6.02±0.41) cm), and the difference was statistically significant (t=11.20, P<0.01). The DAI of TRAIL^-/- colitis mice was higher than that of WT colitis mice (3.00±0.00 vs. 2.32±0.05), and the difference was statistically significant (t=54.40, P<0.01). The histological score of TRAIL^-/- colitis mice was higher than that of WT colitis mice (6.19±0.25 vs. 3.87±0.22), and the difference was statistically significant (t=27.87, P<0.01). Under the microscope, colonic mucosal epithelial injury, crypt structure destruction and inflammatory cell infiltration were more obvious in TRAIL^-/- colitis mice than in WT colitis mice. The alpha diversity of colonic flora was more significant in TRAIL^-/- colitis group compared with that of WT colitis group. At the family level, the relative richness of Deferribacteraceae, Ruminococcaceae, Rikenellaceae, F16 and Paraprevotellaceae significantly increased in TRAIL^-/- colitis group, but the relative richness of Enterococcaceae obviously reduced ((19.839±19.991)% vs. (7.224±11.241)%, (3.564±2.543)% vs.(2.861±3.821)%, (0.123±0.066)% vs. (0.068±0.049)%, (0.032±0.033)% vs. (0.006±0.011)%, (0.153±0.098)% vs. (0.062±0.054)% and (0.013±0.027)% vs. (0.054±0.121)%, respectively; U=51, 69, 53, 35, 49 and 69, respectively; P<0.01 and 0.05, respectively). In addition, at the genus level the relative richness of Oscillospira, Mucispirillum and Cytophaga in TRAIL^-/- colitis group remarkably elevated, and the relative richness of Enterococcus significantly decreased ((2.363±2.147)% vs. (1.813±2.847)%, (19.839±19.991)% vs. (7.223±11.241)%, (0.104±0.153)% vs. (0.046±0.069)% and (0.076±0.049)% vs. (0.135±0.074)%, respectively; U=70, 51, 66 and 65, respectively; P <0.05 and 0.01, respectively). Conclusion TRAIL deficiency aggravate DSS-induced colitis, and increase the alpha diversity of colonic microbiota in colitis mice.