1.Pathogenic effect of intestinal immune dysfunction on irritable bowel syndrome in mouse model
Donghui CHEN ; Hairong CHEN ; Zhiqiang LIU ; Pengyuan ZHENG
Chinese Journal of Microbiology and Immunology 2013;(6):445-451
Objective To investigate the role of intestinal immune dysfunction in the pathogenesis of irritable bowel syndrome(IBS) and to study the effects of Clostridium butylicum on the regulation of intestinal immune disorders.Methods A total of 50 male 6-week-old C57BL/6 mice were randomly divided into three groups,including the experimental group (n =20),the control group (n =20) and the Clostridium butylicum group(n =10).A mouse model of constipation-predominant IBS (C-IBS) was established by perfusing sodium butyrate solution(200 μl,concentration of 500 mmol/L) into the mouse colon twice a day for three consecutive days.The mice in control group were intrarectally perfused with normal saline enema (200 μl).Two hours before the perfusion of sodium butyrate into colon,the mice in Clostridium butylicum group were given Clostridium butylicum 500 μl(viable cell concentration of 1×109 CFU/ml) by oral gavage once a day for six days.The colorectal distention test(CRD) was carried out for evaluation of clinical parameters.HE staining of intestinal tissue section was performed for histopathological assessment of colonic mucosal inflammation.Intestinal intraepithelial lymphocytes (IELs) and lamina propria mononuclear cells (LPMCs) were isolated and analyzed by flow cytometry to evaluate the correlation between IBS and intestinal immune dysfunction/abnormal activation of intestinal immune cells in mouse model of C-IBS,and to assess the regulatory effects of Clostridium butylicum on the intestinal immune disorder.Results (1) Compared with the control group,the mice in experimental group showed a significant change in physiological parameters,histological structure of colon,inflammatory cells infiltration and low-grade inflammatory state.There was a significant increase in scores of CRD and a decrease in lowest sensory threshold (t=8.926 and t=6.103,both P<0.001) ; (2) There was a decrease in the numbers of DC in IELs (t =2.878 and t =3.086,both P<0.05),but an increase in the numbers of macrophage (t=3.191,P<0.05) and the memory T cells in mice with IBS (t=3.071,P<0.05) as compared with that in control group; (3)DCs were decreased (t=2.880 and t=2.664,both P<0.05),but memory T cells were increased (t =3.732 and 2.682,P<0.01 and P<0.05) in the LPMCs of mice in experimental group; (4)There was no significant difference in the physiological index between the mice in control group and the Clostridium butylicum group.Levels of memory T cells,macrophages and DCs in the IELs were close to the normal level (6 d,t =1.103,0.0213,0.418,all P>0.05),and levels of macrophages and DCs in the LPMCs of mice in the Clostridium butylicum group were also similar to that in the control group (6 d,t =0.782,0.347,both P>0.05) ; (5) Compared with the mice in experimental group,the level of memory T cells in LPMCs of mice treated with Clostridium butylicum was dramatically declined (6 d,t=2.346,P=0.0470,P<0.05),however,which was still higher than that of mice in control group (6 d,t =2.233,P =0.0476,P<0.05).The intestinal immune function was restored to normal level with Clostridium butylicum intervention.Conclusion The pathophysiologic mechanism of IBS might be closely related to the abnormal activation of intestinal immune cellsand disordered functional state in the intestinal mucosa.Clostridium butylicum could regulate the intestinal immune homeostasis and restore the physiological function of gastrointestinal tract.
2.Effects of aryl hydrocarbon receptor on Th17 cells activation in a murine model of irritable bowel syndrome
Yuying WANG ; Donghui CHEN ; Pengyuan ZHENG ; Fuguang LI
Chinese Journal of Microbiology and Immunology 2014;(4):299-303
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 .
3.Effects of Clostridium butyricum combined with glutamine on the expression of intestinal tight junction protein in stress mouse model
Donghui CHEN ; Yong YU ; Zhiqiang LIU ; Fuguang LI ; Pengyuan ZHENG
Chinese Journal of Digestion 2014;34(8):544-549
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.
4.Effects of Bifidobacterium infantis on intestinal corticotropin releasing factor receptors and mast cells activation in a murine model of irritable bowel syndrome
Simeng LIU ; Pengyuan ZHENG ; Fuguang LI ; Donghui CHEN
Chinese Journal of Microbiology and Immunology 2014;(8):571-576
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 .
5.Effects of treatment with hydroxythy starch on the serum albumin and immunologic function in postoperative patients with obstructive jaundice
Anyin DAI ; Pengyuan XU ; Dali SUN ; Yunyun CEN ; Xiongzhi CHEN ; Jian LIU ; Weiming LI ; Yuxing QI
Parenteral & Enteral Nutrition 2009;16(4):215-218
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.
6.Hand hygiene management among health care workers in stomatology medical institutions in Zunyi
Xia ZHU ; Ye XIAO ; Shaowei ZHANG ; Pengyuan LEI ; Weiwei HAO ; Pengyi GUO ; Chen KAN
Chinese Journal of Infection Control 2016;15(8):570-572
Objective To understand the current management situation of hand hygiene (HH)among health care workers(HCWs)in stomatology medical institutions in Zunyi,and provide basis for the scientific management of HH.Methods In October-December 2014,the on-site observation and questionnaire survey were adopted to investigate current management situation of HH in 127 stomatology medical institutions in Zunyi.Results The rates of qualified installation of sink,faucet,detergent,alcohol-based hand rub,and hand-drying facilities were 60.63%, 11 .02%,92.13%,22.05%,and 37.80% respectively.Except detergent and sink,the installation of other HH supplies in stomatology medical institutions in Zunyi were not ideal.The correct rate of HH among HCWs was 33.07%(42/127),qualified rates of HH methods among HCWs in departments of stomatology,comprehensive out-patient departments, stomatological clinics,and teeth-cleaning clinics were 50.00%,23.08%,23.53%,and 0 respectively.Conclusion HH facilities are not perfect in primary stomatology medical institutions in Zunyi,health ad-ministrative departments’supervision on HCWs’HH is inadequate,the awareness rate of HH knowledge and com-pliance to HH among HCWs are both low.
7.The study of oral immunization with the transgenic tomato anti-caries vaccine in SD rats
Yu GU ; Jianguo LIU ; Guohui BAI ; Jiayuan WU ; Xiaoyan GUAN ; Zhu CHEN ; Pengyuan BAI ; Jing ZHAO
Chongqing Medicine 2015;(23):3169-3171,3175
Objective SD rats were immuned with the transgenic tomatoes which carried fused gene of a region of PAc Strep-tococcus mutants and cholera toxin B subunit.The immunogenicity was tested to explore secure and economic edible vaccines a-gainst dental caries.Methods A total of 18 eighteen-day-old female SD rats were subdivided randomly into three groups:the exper-imental group which were fed with transgenic tomato juice containing chimaera protein PAcP/CTB;the positive control group which were treated with deactivated S.mutans;the negative control group which were not treated with transgenic tomato juice.Rats were immuned once per week for four weeks.Blood and saliva were collected at one day before the first immunity and one week after each immunization.IgG of blood serum and SIgA of saliva were detected using the enzyme-linked immunosorbent assay (indirect ELISA) testing.On day 70,rats were terminated.The maxillary and mandibular bones were subsequently taken out to count dental caries′scores.Results Post immunization,the experimental group and the positive control group had statistical significant levels of speci-ficity IgG in serum and SIgA in saliva compared to the negative control group (P <0.05).There was a significance difference be-tween the experimental group and the negative control group except in Dx levels of caries loss (P <0.05).Conclusion The targeted protein expressed on the transgenic tomatoes is immunogenic,which can effectively induce mucous membrane immune response and the systematical immunoreaction to suppress the occurrence of the dental caries.
8.Early plasma albumin and inflammatory factors changes in patients undergoing abdominal surgery
Shumin LI ; Pengyuan XU ; Yunyun CEN ; Xiongzhi CHEN ; Dali SUN ; Weiming LI ; Yuxing QI ; Yijun LI ; Qingwen XU
Chinese Journal of General Surgery 2010;25(12):1002-1004
Objective To probe the changes of plasma albumin concentration and its correlation with that of blood inflammatory factors at the postoperative early stage in patients undergoing intraabdominal surgery. Methods From August 2008 to March 2009, 45 patients undergoing abdominal surgery were divided into three groups according to different types of operation with 15 cases in each group, cholecystectomy group( A), chole cystectomy plus common bile duct exploration group(B) and radical resection of alimentary duct maliguance group (C). Before the surgery and 12,24,48,72 h after operation, plasma albumin contentserum IL-6 and TNF-α concentration were measured. Results Postoperatively the content of plasma albumin did not change significantly in group A ( P > 0.05 ), while that decreased after operation in group B and group C(P <0.01 ). The postoperative concentration of serum IL-6 and TNF-α increased in group A at 12, 24 h and 48 h after operation(P <0.01 ). In group B and group C IL-6 and TNF-α increased at all tested time point after operation ( P < 0.01 ). The postoperative alterations of IL-6 and TNF-α were statistically different between the three groups at all time points(P <0.01 ). The content of plasma albumin was in a negative correlation with the concentration of IL-6 and TNF-α; ( r = - 0.376, P = 0.000; r =-0.772,P = 0.000). Conclusions The content of plasma albumin decreased at the early stage after major and moderate abdominal surgery. The content of plasma albumin was in a negative correlation with the concentration of inflammatory factors at the early stage after abdominal surgery.
9.Research progress on risk factors associated with postoperative recurrence in patients with Crohn's disease after bowel resection.
Shanwen CHEN ; Pengyuan WANG ; Yucun LIU
Chinese Journal of Gastrointestinal Surgery 2015;18(1):89-92
Emergency and complications make bowel resection necessary for patients who suffer from Crohn's diseaes (CD) which is traditionally treated mainly by medical therapy. CD patients can obtain temporary relief after bowel resection. However, the high recurrence rate after surgery tends to result in repeated operations. Preoperative factors, the details of the surgery and postoperative medical treatment are considered to be related to postoperative recurrence. In this manuscript, progress on risk factors associated with postoperative recurrence in patients with Crohn's disease after bowel resection is summarized.
Colectomy
;
Crohn Disease
;
Humans
;
Postoperative Period
;
Recurrence
;
Reoperation
;
Risk Factors
10.Research progress of the association of hydrogen sulfide with colorectal cancer and its associated anti-tumor drugs.
Shanwen CHEN ; Pengyuan WANG ; Yucun LIU
Chinese Journal of Gastrointestinal Surgery 2017;20(7):834-840
As the third confirmed gaseous transmitters, hydrogen sulfide was found to play a vital role in the eternal milieu both physiologically and pathologically. What's intriguing is that, there exists a debate about the role of hydrogen sulfide in the pathogenesis of cancer, especially colorectal cancer. In this manuscript, progress of the effect of low and high dose hydrogen sulfide on the tumorigenesis of colorectal cancer and research on the antitumor new drugs stemmed from the function of hydrogen sulfide is summarized. Recently, the "bell-shape" model of the action of hydrogen sulfide illustrated that hydrogen sulfide was able to promote the progression of tumor in low dose and inhibit the progression of tumor in high dose. Based on these results, both hydrogen sulfide synthesis inhibitors and hydrogen sulfide donors(chimeras with NSAIDs) have been widely studied as potential anti-tumor novel drugs. According to the actual results of present studies, hydrogen sulfide synthesis inhibitors featured by aminooxyacetic acid (AOA) is more promising as a novel kind of drug. The future research based on the relationship between hydrogen sulfide and rectal cancer may bring insights into the development of novel anti-tumor drugs, thus producing tremendous potential clinical value.