Objective To detect the expression of CRF and CRF receptors in colonic mucosa of DSS induced colitis in mice model and to study the effect of CRF and CRF receptors on the development.Methods Six to eight weeks healthy female BALB/c mice were divided into control group and DSS group.Setting up DSS colitis model and colitis was evaluated by the disease activity index(DAI) and histological score.The immunofluorescence technique was used to assay the CRF1 and CRF2 receptors expression in colonic mucosa.The expression of CRF and CRF receptors protein were analyzed by western blotting.Results DSS colitis was set up successfully with significant inflammation in colonic mucosa by the disease activity index (DAI) and histological score.Immunofluorescenee staining evidenced that expression of CRF1 receptor in DSS colitis group has no significant deviation compared to control group(P ＞ 0.05),while the expression of CRF2 receptor was elevated in DSS colitis group compared to control group (P ＜ 0.05).CRF2 receptor was localized in epithelial cells and mononuclear cells in the lamina propria.The levels of CRF and CRF2 receptor protein by western blotting were higher in in DSS colitis group compared to control group (P ＜ 0.05).The level of CRF1 receptor protein in DSS colitis group had no significant deviation compared to control group(P ＞ 0.05).Conclusion The higher expression of CRF and CRF2 in colonic mucosa of DSS colitis may participate in the development of colitis.

Objective To investigate the possible influence of Lewis B expression in Helicobacter pylori (H.pylori) on bacterial adhesion property, and determine the relationship between babA 2 gene in H.pylori and peptic ulcer. Methods The bacterial adhesion property in a total of 78 H.pylori strains was performed using adherence assay in vitro, and the frequency of babA 2 gene was determined by polymerase chain reaction (PCR). Results Among 47 H.pylori strains isolated from peptic ulcer patients, 18 (38.3%) were positive for babA 2 gene, while babA 2 was positive in 14 (45.2%) of 31 strains isolated from non ulcer dyspepsia patients ( P =0.427). Among 31 H.pylori strains expressing Lewis B, 24 (77.4%) were positive for adherence assay, compared with 38 (80.9%) of 47 H.pylori strains without expression of Lewis B ( P =0.463). Conclusions (1) There is no association between babA 2 status in H.pylori strain and peptic ulcer in our population; (2) The expression of Lewis B in H.pylori does not interfere with bacterial adhesion property, and thus supports that the gastric epithelium is the receptor for Lewis B of H.pylori.

This study is to investigate the protein and mRNA expressions of pro-inflammatory and anti-inflammatory cytokines in U937 foam cells and effects of Ginkgo biloba extract (GbE) on the cytokines.U937 cells were cultured with different concentrations of GbE (0.1,1,and 10 μg·L-1),and stimulated by 100 mg·L-1 oxidized low density lipoprotein (ox-LDL) for 24 h.The expressions of interleukin-1β (IL-1β),tumor necrosis factor-α (TNF-α) and interleukin-10 (IL-10) in culture solution were detected by enzyme-linked immunosorbant assay (ELISA) and reverse transcriptase polymerase chain reaction (RT-PCR).The results showed that incubated with 100 mg·L-1 ox-LDL for 24 h,the U937 cells became foam cells,the protein or mRNA expressions of IL-1β,TNF-α,IL-10,and its receptor IL-10R in U937 foam cells were higher markedly than those in normal U937 cells.When the cells were pretreated with GbE (0.1,1,and 10 μg·L-1),the increases of IL-1β and TNF-α in U937 foam cells were remarkably inhibited,but IL-10 expression increased greatly.Especially when cells were pretreated with 10 μg·L-1 GbE,the protein and mRNA expressions of IL-1β and TNF-α were markedly lower than those in U937 foam cells.The protein expression of IL-10 and mRNA expressions of IL-10 and its receptor IL-10R were markedly higher than those in U937 foam cells.GbE inhibited production of pro-inflammatory cytokines IL-1β and TNF-α,but up-regulated the production of anti-inflammatory cytokine IL-10 and its receptor IL-10R in U937 foam cells,which might be related with its anti-atherosclerotic actions.

Objective To investigate the effect of corticotrophin-releasing factor (CRF) on the regulation of Toll-like receptor 4/NF-κB signaling pathway expression in human intestinal epithelial cell line HT-29. Methods HT-29 cells were divided into four groups, normal control group, LPS group (LPS 20 μg/ml stimulated for 24 h), CRF group (CRF 20 ng/ml stimulated 24 h) and CRF+ LPS group (CRF incubated for 12 h then changed to LPS for another 12 h). After stimulation, the expression of TLR4 mRNA of each group was examined by reverse transcriptase polymerase chain reaction (RT-PCR). Total cell protein were extracted and the expression of TLR4 and NFκB p65 at protein level were detected by western blotting.Cell culture supernatant was collected and the secretion of interleukin-8 was detected by enzyme-linked immunoasorbent assay (ELISA). Results The expression of TLR4 in LPS group at mRNA and protein level were 0.31±0.04 and 0.48±0.17,there was no significant difference compared with normal control group (0.28±0.02 and 0.45±0.12,t＝0.216 and 0.712 , P＞0.05 ). In CRF group which were 1.05±0.06 and1. 08±0.21, significantly higher than normal control group (t＝3.721 and 3.802, P＜0.05). In CRF+LPS group which were 1.68±0.05 and 1.81±0. 18,significantly higher than CRF group (t＝4. 816 and 3. 918, P＜0.05).The results of NF-κB p65 expression at protein level and interleukin-8 expression of cell culture supernatant were consistent with the results of TLR4 expression at mRNA and protein level.Conclusion CRF not only activate TLR4/NF-κB signaling pathway in human intestinal epithelial cell,but enhance the reaction of intestinal epithelial cell to LPS as well, which resulting in increased interleukin-8 secretion.

Objective To investigate the effects of Bifidobacterium infatn ison the expression of in -testinal corticotropin releasing factor ( CRF) receptors and how the peripheral CRF receptors activate mast cells in a murine model of irritable bowel syndrome (IBS).Methods Thirty BALB/c male mice were ran-domly divided into three groups including control group , model group and Bifidobacterium infantis group. The mouse model of IBS was established by using chronic restraint stress .Mice in Bifidobacterium infantis group received daily intragastrical administration of Bifidobacterium infantis for 14 days.Mice in control and model groups were treated with equal volume of saline .Then all mice were killed after the assessment of weight and abdominal withdrawal reflex ( AWR) .The levels of histamine , tryptase and tumor necrosis fac-tor-α( TNF-α) in serum samples were detected by ELISA .The expression of CRF in colonic mucosa was analyzed by immunohistochemistry .The expression of CRF-R1 and CRF-R2 in mast cells and the number of mast cells in colonic mucosa were detected by double immunofluorescence staining assay .The expression of CRF-R1 and CRF-R2 at mRNA level in colon were detected by reverse transcription polymerase chain reac-tion ( RT-PCR) .Results Compared with control group , the levels of histamine , tryptase and TNF-αin pe-ripheral blood samples , the expression of CRF-R1 and CRF-R2 at mRNA level , and the number of mast cells, CRF-R1+mast cells and CRF-R2+mast cells in colonic mucosa were increased significantly in model group (P<0.05), but were remarkably down-regulated with the treatment ofB ifidobacterium infantis (P<0.05).Conclusion Bifidobacterium infantis could reduce the activation of mast cells in a murine model of IBS by inhibiting the expression of CRF-R1 and CRF-R2 in intestinal mast cells .

Objective This experiment aimed to study the influence of deep sea water (DSW) on wound healing of mice .Methods 24 Mice were randomly divided into two groups :group DSW(n=12) and group sterilize tap water(STW)(n=12) ,freely feeding for 14 days respectively ,and calculated the amount of food and water .On the 15th day ,1 cm × 1 cm size of wound was established on the back area of mice ,and continued to feed with DSW and STW respectively .Tracking the wound healing rate .Specimen was taken in the edge of wound tissue on postoperative 3 ,5 ,7 days ,then observed histopathological changes .Results Compared group DSW with group STW ,there was no significant difference in the total amount of food and water .5 days after the formation of wounds ,the wound healing rate of group DSW was significantly higher than group STW .Histological observation :compared with group STW ,vascular endothelial cells and new capillaries of the group DSW was increased ,and group DSW had less inflammatory cell and more fibroblast cells proliferation .Conclusion deep sea water can promote wound healing .

Objective To detect the dynamic Th17 cells in a murine model of irritable bowel syn-drome ( IBS) and to study the effect of aryl hydrocarbon receptor ( Ahr) on Th17 cells activation .Methods Thirty BALB/c male mice were randomly divided into three groups including experiment group ,control group and Ahr antagonist group .A murine model of IBS was established by perfusing three nitrobenzene sulfonic acid (TNBS) into the colon of mice.Equal volume of saline was used to set up the control .The mice in Ahr antagonist group were intraperitoneally injected with 10 μg Ahr antagonist for four consecutive days .All mice were evaluated for visceral hypersensitivity and colonic mucosal inflammation .Mesenteric lymph nodes (MLNs) and peripheral blood mononuclear cells (PBMCs) were detected by flow cytometry through staining Th17 cells.The distribution of Ahr and IL-17A in colon and the number of Th17 cells activated by Ahr (Ahr and IL-17A double positive ) were detected by double immunofluorescence staining .Results ( 1 ) The percentage of Th17 cells in MLNs was significantly increased in experiment group followed by those in Ahr antagonist group and control group (P<0.05).(2)Compared with control group,the number of Th17 cells in peripheral blood samples was significantly increased in experiment group and Ahr antagonist group ( both P<0.05 ) ,but there was no difference between Ahr antagonist group and experiment group ( P=0.642 ) .( 3 ) The number of Ahr-activated Th17 cells ( Ahr+IL-17A+) was significantly increased in experiment group (10.00±1.58) as in comparison with that in control group (3.80±0.83,P<0.05),but the number was de-creased with Ahr antagonist intervention ( 5.80 ±0.83 , P<0.05 ) .Conclusion The number of activated Th17 cells was increased in MLNs and peripheral blood samples from mice with IBS .Ahr played an important role in the activation of Th17 cells in intestines.However,the number of Ahr-activated Th17 cells in intestinal mucosa and the proportion of Th 17 cells in MLNs could be down-regulated through blocking Ahr .

Objective To investigate the mechanism of the permeability of intestinal mucosa in the pathogenesis of irritable bowel syndrome (IBS) and the interventional effects of Clostridium butyricum combined with glutamine.Methods According to random number method,fifty BALB/c mice were divided into control group,experimental control group,glutamine group,Clostridium butyricum group and combination group.IBS mice model was established by water-avoidance stress (WAS) experiment.The defecating time of mice and fecal water content were detected by dyed stool after mice gavaged with methylcellulose (1.5％).The pathological injury of intestine was assessed by hematoxylin and eosin staining.The visceral sensitivity was evaluated by colorectal distention test (CRD).The changes of the permeability of intestine was evaluated by detecting the changes of serum D-lactic acid (D-LA),level of diamine oxidase (DAO),expressions of intestinal epithelial cells (IEC) cell tight junction protein (TJ) (occludin-1,claudin-1,zonula occludens-1 (ZOL-1)) at protein level.The interventional effects of Clostridium butyricum combined with glutamine were evaluated.t test was performed for comparison between groups,and analysis of variance was used for comparison among multi-groups.Results Compared with the control group,the defecating time of experimental control group was significantly shorten ((100.40±14.80) min vs (75.88±12.20) min and water content of fecal significantly increased ((54.76±9.98)％ vs (74.95±7.15)％,t =3.692 and 4.023; P=0.002 and 0.002).The lowest threshold of visceral sensitivity significantly decreased ((40.87 ± 4.82) mmHg (1 mmHg=0.133 kPa) vs (27.80±3.18) mmHg; t=8.761,P＜0.01),while the mucosal pathological injury score significantly increased (0.50±0.15 vs2.60±0.97; t＝6.034,P＜0.01).The level of D-LA ((1 476±246.8) ng/L vs (913.6±90.1) ng/L)) and DAO ((3 391.0±256.9) vs (5 096.0±725.2) ng/L) significantly increased (t=40.920 and 29.810; both P＜0.05),and the expression of tight junction protein ZOL-1 (0.165±0.005 vs0.119±0.003),occludin-1 (0.104±0.016 vs 0.022±0.006) significantly decreased (t=19.830 and 19.830; both P＜0.01).Compared with the experimental control group,after intragastric intervention the defecating time of glutamine group,Clostridium butyricum group and combination group increased ((90.50±3.78),(97.56±8.79) and (99.89±11.90) min and water content of fecal decreased ((69.33±6.71)％,(58.07±8.97)％ and (56.74±8.12)％) and the differences were statistically significant (F=10.020 and 8.740; both P＜0.01).The results of Clostridium butyricum group and combination group were good (F=2.481 and 4.874; both P＜0.05).And the lowest threshold of visceral sensitivity significantly increased ((31.80±2.69),(36.04±5.06) and (38.93±3.30) mmHg; F=2.420,P＜0.05),the result of combination group was the best (F=3.550,P＜0.01).Jejunal mucosal injury was significantly reduced (2.00 ± 0.94,1.30 ± 0.68 and 1.30±0.48; F＝11.350,P＜0.01).After intragastric intervention,serum levels of D-LA ((1 370.0± 78.9),(1 066.0±155.5) and (1 039.0±129.0) ng/L) and DAO ((4 808.0±477.4),(3 713.0± 595.0) and (3 725.0±615.9) ng/L) of glutamine group,Clostridium butyricum group and combination group significantly decreased (F＝37.480 and 27.670; both P＜0.01).The level of ZOL-1(0.126± 0.014,0.125±0.006,0.138±0.004) and occludin 1 (0.037±0.013,0.073±0.028,0.078±0.027) of glutamine group,Clostridium butyricum group and combination group significantly increased,and the differences were statistically significant (F=5.867 and 10.630; both P＜0.05).The change of ZOL-1 of combination group was more than that of Clostridium butyricum group (t =5.457,P ＜ 0.05).Conclusions WAS experiment can induce visceral hypersensitivity,increase the permeability of intestine and reduce the function of intestinal epithelial barrier.Clostridium butyricum and glutamine are effective in the recovery of visceral hypersensitivity and the permeability of mucosal epithelia cells.

Objective To investigate the diagnosis and treatment of congenital mesenteric hiatal hernia in aduls.Methods The retrospective cross-sectional study was conducted.The clinical data of 11 adult patients with congenital mesenteric hiatal hernia who were admitted to the First Affiliated Hospital of Henan University from January 1999 to January 2016 were collected.All patients underwent abdominal X-ray and ultrasound examinations.Patients diagnosed as with intestinal obstruction or suspected intra-abdominal hernias underwent abdominal CT examination,and then were finally confirmed during surgery.Patients diagnosed as with mesenteric hiatal hernia received necrotic tissues resection and tissue repair (small intestine resection and anastomosis) if there was necrosis of hernia contents,and closing mesenteric hiatus.Patients without small intestine necrosis received closure of mesenteric hiatus after retraction of the hernia contents.Observation indicators:(1) clinical manifestations,(2) imaging findings,(3) treatment,(4) pathological examination,(5) follow-up situations.Follow-up using outpatient examination and telephone interview was performed to detect the postoperative complications up to March 2017.Results (1) Clinical manifestations:all 11 patients were acute onset,with incentives of satiation,postprandial exercise and diarrhea.The time from onset to admission was 2.0-30.0 hours,with an average time of 9.8 hours.The main symptoms included abdominal pain,nausea and vomiting,exhaust reduction and other intestinal obstruction performances.Eleven patients received physical examination,and 10 showed abdominal bulge,including 9 with intestinal type.Eleven patients had abdominal tenderness,and 9 combined with rebound tenderness.Abdominal percussion of 11 patients showed hyperresonant without shifting dullness,and active,muted and fading bowel sounds were detected in 1,3 and 7 patients,respectively.(2) Imaging examination:of 11 patients receiving abdominal X-ray examination,2 had intestinal loop and 4 had the intestinal obstruction performances such as typical gas-liquid plane.Abdominal ultrasound examination of 11 patients showed no specific findings due to abdominal intestinal gas,and 10 with peritoneal effusion.Of 11 patients,1 didn't receive abdominal CT scan due to preoperatively misdiagnose with acute appendicitis and 10 underwent abdominal CT scan.Nine patients were diagnosed with intestinal torsion by abdominal CT scan and then underwent enhanced CT scan,and 8 with small mesenteric vascular torsion and swirling sign were diagnosed with small intestine torsion and partial necrosis of small intestine.(3) Treatment:1 patient preoperatively misdiagnosed with acute appendicitis was converted to exploratory laparotomy,and 10 patients underwent exploratory laparotomy due to complete intestinal obstruction or progressive increase in symptoms.Intraoperative exploration showed that intestinal mesenteric hiatus and colon mesenteric hiatus were respectively in 8 and 3 patients,and hiatuses were round or oval,with a diameter of 2.0-8.0 cm and an average of 4.4 cm.Hernia contents were small intestine.The partial small intestine in 10 patients were resected and then mesenteric hiatus was closed due to necrosis of the small intestine,with removal length of 110-250 cm and an average of 176 cm,and length of remaining small intestine was 80-230 cm,with an average of 159 cm.The hernia into small intestine in 1 patient without complete necrosis was retracted to abdominal cavity after symptomatic treatment,and closing mesenteric hiatus.Eleven patients were cured and out of hospital after operation,without nosocomial complications.(4) Pathological examination:small intestine ischemic necrosis was detected in 10 patients after partial small intestine resection.(5) Follow-up situations:all patients were followed up for 12-24 months,without malnutrition,short bowel syndrome and other complications.Conclusions Without history of abdominal trauma or surgery,with incentives of the satiation,postprandial exercise and diarrhea,abnormal retroperitoneal small intestine shadow and small intestinal torsion diagnosed by CT scan and absent intestine sign by enhanced CT scan can be helpful to diagnose congenital mesenteric hiatal hernia in adults and small intestinal necrosis.Surgery is the only effective method in the treatment of congenital mesenteric hiatal hernia in adults.

Objective: To investigate the effects of trentment with hydroxythy starch(130/0.4) on the serum albumin and immunologic function in postoperative patients with obstructive jaundice.Methods: 24 patients with obstructive jaundice were randomly divided into the control group(n=12) and hydroxythy starch(130/0.4) (HS) group(n=12). The serum ALB was detected 1d, 3d, 5d, 7d after the operation,and the immunologic index were detected 1d and 7d after the operation.Results: The ALB content of control group and HS group were not significantly different in the postoperative 1d and 7 d(P>0.05) and had significant differences in the postoperative 3 d, 5 d (P<0.05).All immunologic index had no significant differences in the postoperative 1d, 7d (P>0.05).Conclusion: The ALB content of patients with obstructive jaundice may decrease postoperatively. Treatment with hydroxylthy starch(130/0.4) can alleviate the drop of the ALB content. But it has no effects on immunologic function.