Long noncoding RNAs( lncRNAs)are a class of non-protein-coding RNA species that are greater than 200 nucleotides in length. LncRNAs regulate gene expression at the epigenetic,transcriptional and post-transcriptional levels, and are widely implicated in the physiological and pathological processes. Recent evidences indicated that lncRNAs play important roles in the development and progression of many digestive system diseases. Studies focusing on lncRNAs may provide valuable data for the prediction,diagnosis,therapeutics and prognosis assessment for these diseases. In this article,the fundamental principles and progress in study on lncRNAs in digestive system diseases were summarized.

AIM:To analyze the expression profile of long non-coding RNA (lncRNA) in the liver tissues of drug-induced liver injury ( DILI) and immune liver injury ( ILI) .METHODS:The technique of lncRNA microarray was used to inspect the lncRNA expression profile in the mouse liver tissues that the liver injury was induced by acetaminophen or concanavalin A .The raw data of lncRNA were pretreated for normalization .RESULTS:Compared with normal hepatic tissue, the lncRNA which had more than 1.5-fold variation and significant difference (P<0.05) by statistical analysis were regarded as lncRNA with differential expression .A total of 68 lncRNA with differential expression were found in the hepatic tissues of DILI, with 21 increased more than 1.5 folds and 47 reduced more than 1.5 folds.A total of 60 lncRNA with differential expression in the liver tissues of ILI were observed , with 17 increased more than 1.5 folds and 43 reduced more than 1.5 folds.In all lncRNA, 8 was simultaneously up-regulated in 2 liver injury models , accounting for 38%and 47%respectively, while 28 was simultaneously down-regulated in 2 liver injury models, accounting for 59%and 65%re-spectively .CONCLUSION:lncRNA expression profiles of DILI and ILI change significantly in comparison with normal hepatic tissue , and there are also differences between 2 hepatic damage models .The simultaneous changes of lncRNA may participate in the same or similar pathophysiological process , while the differences may be involved in relatively particular mechanisms .

Objective To observe the dynamic changes of leptin and transforming growth factor (TGF)-β1 in formation of liver fibrosis in rats,and the role of leptin in liver fibrosis.Methods Six rats were served as normal control(0 week)and killed at the beginning of the study.Thirty-two rats were subcutaneously injected with 60%CCl4 to establish fibrotie models.The rats were than sacrificed at 3,6,9 and 12 weeks with eight each.Expressions of leptin mRNA,Ob-Rb mRNA,TGF-β1 mRNA and TGF-βRⅡmRNA in liver were detected by reverse-transcription polymerase chain reaction(RT-PCR).Expression of Smad3 and Smad7 proteins were determined by Western blot.The expression and loealization of leptin,Ob-Rb and TGF-β1 in liver tissue were detected by immunohistochemistry.Results The expression of leptin mRNA,Ob-Rb mRNA,TGF-β1 mRNA and TGF-βR Ⅱ mRNA in 0 week were 0.43±0.45,0.57±0.21,0.19±0.12 and 0.30±0.22,respectively,and increased to 1.27±0.19,1.70±0.29,1.70±1.00 and 2.14±1.02 at 12 weeks(P＜0.05).At the same times the level of Smad3 increased from 0.44±0.24 at 0 week to 1.75±1.05 at 12 weeks.While the expression of Smad7 was decreased from 1.35±0.12 at 0 week to 0.74±1.21 at 12 weeks(P＜0.05).The expressions of leptin,Ob-Rb and TGF-β1 were increased with the formation of the fibrosis(P＜0.01).Conclusions With the development of liver fibrosis,the expressions of leptin,Ob-Rb mRNA and protein are increased and improved the formation of the fibrosis.Its mechanisms may be correlated with up-regulation of TGF-β1,TGF-βR Ⅱ and Smda3 expressions and down-regulaton of Smad7 expression by leptin.

Objective To observe the effect of compound fetal liver extract combined with antiviral therapy on liver fibrosis in patients with cirrhosis.Methods 60 patients with liver cirrhosis from April 2014 to July 2015 were randomly divided into treatment group and control group(n=30).The patients in control group were treated with conventional anti viral therapy , 30 cases in treatment group were treated with compound fetal liver extract combined with antiviral therapy postoperative.Results 1 month after treatment, the treatment group serum liver fibrosis HA, PCM value respectively(107.5 ±17.8,99.8 ±14.9)ng/mL, were lower than in the control group (138.4 ±15.2,124.1 ±18.1)ng/mL(P<0.05).1 month after treatment, the treatment group liver fibrosis collagen type IV, III procollagen value respectively(58.9 ±11.0,109.2 ±11.1)μg/L, were lower than in the control group (85.7 ±11.2,122.7 ±11.3)μg/L(P<0.05).Conclusion Compound fetal bovine liver extract combined with anti-viral therapy in patients with cirrhosis has good, better than the use of antiviral drugs alone.

AIM: To observe the effect of Tripterygium glycosides on NOXs-ROS-NLRP3 inflammatory signa-ling pathways in the colon tissue in dextran sulphate sodium (DSS)-induced ulcerative colitis (UC) mice, and to investi-gate the underlying mechanisms.METHODS: BALB/c mice were used and the mouse model of UC was established by DSS induction.The mice were randomly divided into 5 groups (model group, low-, medium-and high-dose Tripterygium glycosides groups, and normal group).The colon tissues were collected 21 d after Tripterygium glycosides gavage.The mRNA expression of NLRP3, ASC and caspase-1 in the colon tissues was detected by real-time PCR.The caspase-1 ex-pression in the colorectal mucosa was observed by immunohistochemical method.ELISA was used to detect the protein le-
vels of IL-1α, TNF-αand IL-13.The production of reactive oxygen species (ROS) was measured by chemiluminescence technique, and the consumption rate of NADPH, which was inhibited by DPI, was analyzed to determine the activity of NADPH oxidases (NOXs).The neutrophils were isolated, and the ROS production, NOXs activity, and the mRNA ex-pression of NLRP3, ASC and caspase-1 were also detected.RESULTS: The colon tissues were abnormal with different de-grees in Tripterygium glycosides groups, and histopathological scores were lower than that in model group.In Tripterygium glycosides groups, in addition to the mRNA expression levels of caspase-1 in the colon tissues between normal group and high-dose group, ROS production, NOXs activity and the mRNA expression levels of NLRP3, ASC and caspase-1 in the colon tissues and colon-isolated neutrophils were lower than those in model group (P <0.05), and higher than those in normal group (P <0.05).The results of pairwise comparison for the efficacy of Tripterygium glycosides administration showed that the above indexes were statistically significant except the mRNA expression levels of caspase-1 between middle-dose group and high-dose group.Tripterygium glycosides administration significantly decreased the expression levels of proinflammatory cytokines IL-1αand TNF-αin the homogenates of colon tissues in the model mice (P <0.05).No differ-ence of IL-13 expression among the groups was observed.CONCLUSION: Tripterygium glycosides inhibits NOXs-ROS-NLRP3 inflammatory signaling pathways to reduce the expression of IL-1α, TNF-αand other proinflammatory cytokines, and attenuates DSS-induced ulcerative colitis in mice, by which the neutrophils might be involved in the process.

Background:Tripterygium glycosides(TG)is effective for treatment of ulcerative colitis(UC)in clinical practice, however,the underlying mechanism has not been clarified yet. Aims:To investigate the therapeutic effect of TG on dextran sulfate sodium(DSS)-induced experimental colitis in mice and its possible mechanisms. Methods:Sixty healthy male BALB/ c mice were randomly divided into six groups:model control group,low,medium and high-dose TG group,blank control group and normal control group. Mice in the first four groups drank 5% DSS freely for 7 days to induce experimental colitis;simultaneously,distilled water,9. 01,27. 03 or 81. 09 mg/(kg·d)TG were given intragastrically for 21 days in these four groups,respectively. Histopathological changes of colonic mucosal tissues were observed;expressions of TLR4 mRNA and protein were determined by RT-PCR and Western blotting;expression of NF-κB p65 was detected by immunohistochemistry;concentrations of IL-1α,TNF-α and IL-13 were measured by ELISA. Results:Tissue damage and inflammation in varying degrees were observed in colonic mucosal tissues in TG groups with different dosage,but all were less severe than those in model control group. Expressions of TLR4 mRNA,TLR4 protein,and NF-κB p65 in colonic mucosal tissues,as well as concentrations of IL-1α and TNF-α in supernatant of colonic homogenate were significantly lower in TG groups than those in model control group(P < 0. 01). These parameters in medium and high-dose TG groups were significantly lower than those in low-dose TG group(P < 0. 05),but higher than those in blank control group and normal control group(P < 0. 05). Except for TNF-α,no significant differences were seen between medium and high-dose TG groups(P > 0. 05). Conclusions:TG exerts a protective effect on DSS-induced experimental colitis in mice. The underlying mechanism of its anti-inflammatory effect might be related with the inhibition of TLR4 / NF-κB signaling pathway activation and subsequently suppressing downstream proinflammatory cytokines expression and secretion.

AIM: To observe the changes in expression and activity of the transcription factor myocyte enhancer factor 2A (MEF2A) during hepatic stellate cells (HSC) activation, and to study the roles of MEF2A in the process of HSC activation. METHODS: Cultured HSC was isolated from male sprague-dawley rat liver on plastic dishes and were used as model of activation. The freshly isolated (0 day) and cultured HSC at time points of 1st, 2nd, 3rd, 4th, 5th, 6th, 7th and 8th day were collected. Expression of MEF2A mRNA was detected by real-time quantitative PCR. MEF2A and α-smooth muscle actin (α-SMA, a marker for activated HSC) were tested by Western blotting. Meanwhile, the MEF2A DNA binding activity was determined by electrophoretic mobility shift assays (EMSA). RESULTS: The expression of MEF2A mRNA was small amounts in the freshly isolated HSC and increased gradually after culture on plastic dishes. Western blotting revealed that the freshly isolated HSC expressed very low levels of MEF2A and α-SMA. The proteins of MEF2A and α-SMA were increased gradually in the process of HSC activation. Increased MEF2A protein was correlated with α-SMA. EMSA revealed that MEF2A DNA binding activity was increased gradually during HSC activation. CONCLUSION: In the process of HSC activation, expression and activity of MEF2A are increased gradually, indicating a role in HSC activation.

Objective To investigate the relationship between the expression of NOD-like receptor protein 3 (NLRP3) inflammasome in colonic mucosal tissues of ulcerative colitis (UC) patients and UC mice model and the activity of UC.Methods From December 2016 to January 2018,at Department of Gastroenterology,the First Affiliated Hospital of Zhejiang Traditional Chinese Medicine University,60 patients with UC were recruited,of which 15 cases at remission phase,15 cases at mild activity phase,and 15 cases at moderate activity phase,and 15 cases at severe activity phase;and 15 healthy subjects were selected as healthy control group.UC mice models were established by dextran sulfate sodium (DSS).Forty-eight BALB/c mice were divided into 2.5％ DSS group,5.0％ DSS group and 7.5％ DSS group and control group.The colon tissues of UC patients and UC mice models were pathologically scored.The expression of NLRP3,cysteinyl aspartate specific proteinase 1 (caspase-1) and apoptosis-associated speck-like protein containing CARD (ASC)at mRNA level in colon tissues of UC patients and UC mice models were determined by real time fluorescence quantitative polymerase chain reaction (PCR).The expression of NLRP3,caspase-1 and ASC at protein level in colon tissues of UC patients and UC mice models were detected by Western blotting.One-way analysis of variance and SNK-t test were performed for statistical analysis.Results The histopathological scores of colon tissues of UC patients at remission phase,at mild activity phase,at moderate activity phase,at severe activity phase and healthy controls were 2.37 ± 0.46,4.84 ± 1.29,6.82 ± 0.96,9.42 ± 1.13 and 1.23 ± 0.55,respectively;the differences were statistically significant (F =67.68,P ＜ 0.01).The higher the degree of inflammation,the higher the pathological score,and the differences were statistically significant (all P ＜ 0.05).The pathological scores of colon tissues of mice in the 2.5％ DSS group,5.0％ DSS group and 7.5％ DSS group were 4.54±0.74,6.02± 1.00 and 8.43 ± 1.46,respectively;the higher the dose of DSS,the higher the pathological score,and the differences were statistically significant (all P ＜ 0.05).The expression of NLRP3 at mRNA level of UC at remission phase,mild activity phase,moderate activity phase,severe activity phase and healthy controls were 1.15 ±0.10,1.49 ±0.13,2.00±0.25,2.05 ±0.33 and 0.61 ±0.09,respectively;the expression ofcaspase-1 at mRNA level were 1.13 ±0.08,1.51 ±0.19,2.10 ±0.23,2.88 ±0.33 and 0.61 ±0.11,respectively;the expression of ASC at mRNA level were 1.12 ± 0.08,1.88 ± 0.33,2.53 ± 0.22,3.20 ± 0.24 and 0.59 ± 0.12,respectively;the differences between groups were statistically significant (F =108.43,63.25 and 105.25,all P ＜ 0.01).The higher the degree of inflammation,the higher the mRNAexpression levels of NLRP3,caspase-1 and ASC,and the differences were statistically significant (all P ＜0.01).The higher the dose of DSS,the higher the protein expression levels of NLRP3,caspase-1 and ASC at mRNA level.The higher the degree of inflammation,the higher expression of NLRP3,caspase-1 and ASC in colon tissues of UC patients.The higher dose of DSS,the higher the protein expression levels of NLRP3,caspase-1 and ASC in colon tissues of mice.Conclusions The expression level of NLRP3 inflammasome is different in different stages of UC,the higher degree of inflammatory activity,the higher the expressie level.It is helpful to evaluate the activity of UC by detecting the expression level of NLRP3 inflammasome.

Background:Early diagnosis and treatment can effectively improve the prognosis of colon cancer.Simple,effective and sensitive screening indicators are of great significance for identification of early cancer and precancerous lesions.L-selectin is a cell adhesion molecule,and podocalyxin(PODXL)is its ligand,both of them play key roles in the development of cancer.Aims:To investigate the expression and significance of L-selectin and its ligand PODXL in colon cancer.Methods:A total of 120 cases of pathological specimens(40 hyperplastic polyp,40 colon adenoma,and 40 colon cancer)from Nov.2020 to Nov.2022 at the Frist People's Hospital of Hangzhou Lin'an District and the First Affiliated Hospital of Zhejiang Chinese Medical University were collected,and 20 cases of normal intestinal mucosal tissue were served as controls.qRT-PCR and immunohistochemistry were used to detect the mRNA and protein expressions of L-selectin and PODXL,respectively.Western blotting was used to determine the expressions of L-selectin and PODXL,and their relations with different clinicopathological parameters of colon cancer were analyzed.In addition,60 serum specimens of colon cancer were collected.ELISA was used to detect serum concentrations of L-selectin and PODXL.Results:Expressions of L-selectin and PODXL mRNA and protein in colon adenoma group were significantly higher than those in normal controls and hyperplastic polyp group(P<0.05),and mRNA and protein expressions of L-selectin and PODXL in colon cancer group were significantly higher than those in normal controls,hyperplastic polyp group and colon adenoma group(P<0.05).Significant differences in protein expressions of L-selectin and PODXL were found in different pathological types,lymph node metastasis,Dukes staging in colon cancer(P<0.05).Expression of L-selectin was positive correlated with expression of PODXL in colon cancer(r=0.855,P<0.001).Serum concentrations of L-selectin and PODXL were significantly lower in the initial group than in the relapse group(P<0.05),and serum concentrations of L-selection and PODXL was significantly lower in the non-metastatic group than in the metastatic group(P<0.05).Serum concentrations of L-selectin and PODXL at 3 months after surgery was significantly lower than 3 days after surgery and before surgery(P<0.05).Conclusions:L-selectin and PODXL may be involved in the development and progression of colon cancer.They are carcinogenic proteins,and their detection could provide reference value for the prevention and early diagnosis of colon cancer,and through early screening of lesion could improve the prognosis of colon cancer to a certain extent.