BacKground:Lymphocyte function-associated antigen-1( LFA-1 ) pIays a cruciaI roIe in the pathogenesis of infIammatory boweI disease by reguIating the activation and function of T ceIIs. Aims:To investigate the effect of LFA-1 deficiency( LFA-1-/-)on differentiation of mice na?ve T ceIIs into Th17 ceIIs in vitro. Methods:LFA-1-/- mice were breeded and the progeny genome DNA was extracted from the taiIs for genotyping by PCR. CD4+CD62L+ na?ve T ceIIs were separated from spIenic mononucIear ceIIs of LFA-1-/- progeny mice and wiId type ( WT ) C57BL/6J mice, respectiveIy,by magnetic-activated ceII sorting( MACS),and then the purity of separated ceIIs was determined. Na?ve T ceIIs obtained were cuItured in different inducing systems[ transforming growth factor-β( TGF-β),TGF-β + interIeukin-6 (IL-6),and TGF-β + IL-6 + IL-23]in vitro for Th17 ceII differentiation;ceIIs in each inducing system were coIIected for anaIyzing the ratio of Th17 ceIIs by fIow cytometry,and the expressions of Th17-specific transcription factor ROR-γt and Th17-specific marker IL-17A were measured by qRT-PCR and ELISA methods. Results:AII fifteen progeny mice were identified as LFA-1-/- genotype. Purity of CD4+CD62L+ na?ve T ceIIs separated by MACS was above 95%. Th17 ceIIs couId be induced by Iow-dose TGF-βcombined with IL-6,and the differentiation ratio was increased obviousIy when IL-23 was added. In inducing system containing TGF-β,IL-6 and IL-23,na?ve T ceIIs from LFA-1-/- mice produced more Th17 ceIIs than those from WT mice(17. 2% ± 1. 4% vs. 5. 7% ± 0. 2%,P<0. 001),expressions of ROR-γt mRNA and IL-17A mRNA were up-reguIated(P<0. 001),and IL-17A concentration in ceII cuIture supernatant was increased(P<0. 01). Conclusions:Deficiency of LFA-1 promotes the differentiation of mice na?ve T ceIIs into Th17 ceIIs in vitro.

Objective:To investigate whether the susceptibility of human colon adenocarcinoma cell SW1116 to lymphokine-activated killer cell (LAK)-mediated lysis could be enhanced by low concentration of sodium butyrate, and the possible involvement of intercellular surface adhesion molecule-1 (ICAM-1) and carcinoembryonic antigen (CEA). Methods:Standard MTT assay was used to evaluate the cytotoxic activity of LAK cells to SW1116 cells, flow cytometric and immunofluorescent techniques were used to determine the expression of ICAM-1 and CEA on tumor cells. Results:Treatment of SW1116 with sodium butyrate leads to an increased resistance to LAK-mediated lysis, accompanied by downregulation of ICAM-1 expression and upregulation of CEA expression. Conclusion: Sodium butyrate inhibits rather than enhance LAK activity against SW1116, probably by changing the expression of ICAM-1 and CEA on target cells, which impair the adherence of effector on target cells.

Objective To study effect of triple viable bacterium of bacillus (small peifeikang) on the intestinal microflora and inflammation factors in patients with infectious diarrhea.Methods From October 2010 to October 2014, 120 patients with infectious diarrhea were chosen as study objects, 30 cases of health examination were selected as healthy control group, All patients were divided into 2 groups:60 patients in small peifeikang group and 60 patients in control group.The therapeutic effect and the change of intestinal microflora and inflammation factors were compared between two groups.Results The effective rate of small peifeikang groupwas 100.00%, while that of control group was 96.67%,the difference had statistical significance (χ2 =5.52,P<0.05).The symptom disappearance time of diarrhea, vomiting and fever had statistical difference between two groups (P<0.05), that of dehydration had no statistical significance.After treatment, in small peifeikang group and control group, the number of bifidobacterium, lactobacillus, and B/E value had decreased significantly (P<0.05), and the number of intestinal bacteriahad increased significantly(P<0.05).The distribution of intestinal flora in small peifeikang group was better than that of control group, and the difference had statistical significance(P<0.05).After treatment, in small peifeikang group and control group, the level of endotoxin, IL -6, TNF-αand CRP had decreased significantly( P <0.05).The extent of decreasing in small peifeikang group was more significant than that of control group, and the difference had statistical significance ( P <0.05 ) . Conclusion The triple viable bacterium of bacillus can promote the recurrence of patients with infectious diarrhea by adjusting the distribution of intestinal microflora, and by its immune regulation function.

Background:Faecalibacterium prausnitzii( Fp) is one of the most abundant bacterium in human intestinal microbiota,and is closely correlated with the process of colitis-associated colorectal cancer(CAC). Aims:To observe the effect of Fp on CAC,and investigate the possible mechanism. Methods:The model of CAC was induced by azoxymethane (AOM)and dextran sodium sulfate( DSS). Fifty-two C57BL/ 6J mice were randomly divided into 4 groups:group A (AOM + DSS),group B(AOM + DSS + Fp),group C(AOM + DSS + Fp supernatant)and group D(control group). All the mice were sacrificed on day 92. DAI was assessed,serum levels of TNF-α and IL-10 were determined by ELISA. HE staining was used to examine the grade of tumor. Expressions of VEGF,COX-2,NF-κB in tumor tissue were measured by immunohistochemistry. Results:The tumorigenesis rates of group A,B,C were 100% ,100% and 77. 8% ,respectively;mainly were high-grade intraepithelial neoplasia. The tumor load in group A was significantly higher than that in group B (P < 0. 01),and the spleen index in group B was significantly higher than that in group C(P < 0. 01). Serum level of TNF-α was significantly lower(P < 0. 05)and IL-10 was significantly higher(P < 0. 05)in group A than that in group B. No significant differences in expressions of VEGF,COX-2,NF-κB were found among group A,B and C. Conclusions:Fp had no obvious effect on the occurrence rate of CAC,and Fp supernatant could decrease the incidence of CAC in mice. Fp and its supernatant could reduce the tumor load via regulating the expressions of TNF-α,IL-10.

Objective To study the effect of nonsteroidal anti-inflammatory drugs (NSAIDs) ,cy-clooxygenase-2 ( COX-2 ) inhibitor Piroxicam on the growth of colorectal cancer cells and to evaluate the preventive significance from COX-2 expression and apoptosis. Methods The cell growth of colorectal adenocar-cinoma cell line SW1116 was measured by MTT (3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyl tetrazolium bromide) assay, and COX-2 protein expression by Immunohistochemistry and Western Blot. Apoptosis was characterized by DNA fragmentation. Results The results showed that COX-2 inhibitor Piroxicam could restrain the proliferation of SW1116, which had positively related to its concentration. Concentration higher than 1. 0mmol/L showed cytotoxic effects. The inhibition of COX-2 by Piroxicam appeared within 12 hours, but COX-2 protein level recovered within 24 hours, its expression had negatively related to the concentration of Piroxicam. Apoptosis was induced in SW1116 culture with Piroxicam higher than 0. 1mmol/L. Conclusion It can be concluded that cell inhibition effect is associated with Piroxicam-mediated cell apoptosis and inhibition of COX-2 protein expression in SW1116 cell, because the effects of Piroxicam have the concentration and time dependence, in further clinical research its dosage and time of medication should be considered in preventing or treating colorectal cancer.

Objective The activity and cyclooxygenase (COX-2) protein expression of colorectal adenoc arcinoma cell treated by deoxycholic acids (DCA) were examined in order to find out the carcinogenesis mechanism of DCA. Methods The proliferation of colorectal cancer cells (SW 1116) was detected by MTT metho d . COX-2 protein expression was measured by immunohistochemistry and Western blo t. Results DCA lower than 100 ?mol/L concentration can stimulate the growth of the adenoca rcinoma cells with effect reversible, while higher concentration shows the long -acting effect of inhibition. DCA of 10,50 and 100 ?mol/L concentration can up -regulate the expression of COX-2 in cells, while only 10 ?mol/L can maintain this effect long than 72 hours. The level of COX-2 protein treated by the late r two concentration descends after 48 hours. Conclusion DCA affects the proliferation of SW1116 in two sides. D CA concentration lower than 100 ?mol/L can promote COX-2 protein expression, w hich may be the mechanism of its carcinogenesis.

Objective Annexin A3 (ANXA3) is a member of the annexin family,As the existing studies suggest,ANXA3 is closely related to tumor genesis,development,invasion,metastasis and prognosis.ANXA3 is down-regulated in prostate and kidney cancer,but it is up-regulated in breast cancer,liver cancer and other tumors.ANXA3 is related to tumor size,staging,lymphatic metastasis and prognosis.Silencing ANXA3 expression can not only inhibit the proliferation and invasion of colorectal cancer and hepatocellular carcinoma cells,but also restrain the migration of breast cancer cells.ANXA3 may also be involved in the regulation and maintenance of hepatocellular stem cells through HIF1a / Notch and JNK signaling pathways.The current studies have shown that ANXA3 can serve as a potential biological marker of tumor diagnosis,prediction of chemotherapy sensitivity,and provide a new target for oncotherapy.

Objective To assess the association of obesity, serum lipid levels and insulin resistance with leptin in Nanjing population. Methods One hundred and eighty eight subjects aged 22～81 years in Nanjing were divided into three groups according to BMI and dignosis criteria of diabetes: normal group, obese group and diabetes group. Their weight, height, waist, abdomen and hip circumferences were measured. Total cholesterol, triglycerides, low density lipoprotein cholesterol, high density lipoprotein cholesterol, apoA, apoB, fasting plasma glucose (FPG), true insulin (TI), immunoreactive insulin (IRI) and leptin were also measured. ISI 1=1/(FPG?TI) and ISI 2=1/(FPG?IRI) as insulin sensitivity indexes were used to analyse the relation between leptin and insulin resistance. Results Fasting serum leptin level was associated with sex (four fold as high in women as that in men). BMI was the second strongest determinant of leptin. Leptin was positively correlated with TI (r=0.32, P

Objectives: To explore the epidemiological distribution of body mass index(BMI),waist to hip ratio(WHR) and abdominal circumference in population of Nanjing,Jiangsu Province. Methods: BMI, WHR and abdominal circumference were measured on 3 445 healthy cases aged 18～90 of Nanjing. Epidemiological distribution of BMI,WHR and abdominal circumference were analyzed according to sex and age. Results:The total mean BMI was( 23.28 ?3.49)kg/m 2, and the BMI in male was higher〔(23.80?3.36)kg/m 2〕 than that in female 〔(22.25? 3.49)kg/m 2〕( t =12.75, P

Objective To study the of effets of hFRNK gene transfected by adenoviral vector on human colon cancer cell invasion induced by gastfin.Methods The subjects were divided into three groups:control,G17 and hFRNK group.The G17 group was treated with 100 umol/L gastrin 17 for 12 h to induce Col0320WT cells.As to hFRNK group,Colo320WT cells were infected by pAdhFRNK(MOI:100)for 48 h after transient transfection of pCR3.1-CAR for 48 h and subsequently treated with gastrinl7 for 12 h,and the control group was untreated Colo320WT cells.Expression of phosphorylated FAK(PY397)were assayed by western blot.FAK(PY397)at lamellipoda was observed with a confocal microscope.The influence of hFRNK on formation of signal complex of FAK-Src-p130Cas-Dockl80 was assayed with coimnlunoprecipitation and immunity blotting.Activity of Rac-GTPase was determined by pull down.Results Phosphorylated FAKTyr397 drastically increased with the induction of gastrin.Compared with that of G17 group,FAK (PY397)expression decreased and little FAK(PY397)was found at lamellipoda,at the same time,the signal complex of FAK-Src-p130Cas-Dockl80 did not form,and the activity of Rac decreased.Conclusion hFRNK gene may block gastrin-induced FAK phosphorylation,prevent formation of the signal complex,prevention and treatment of tumor invasion and metastasis.