The aim of this study was to investigate the effect of interleukin 6 (IL-6) on the contraction of colon longitudinal muscle strips in rats with acute pancreatitis (AP) and its underlying mechanism. Rat AP model was established by combined injection (i. p.) of ceruletide and lipopolysaccharide. The effect of IL-6 on spontaneous contraction of longitudinal smooth muscle strips of rat colon was observed by biological function experiment system. The level of serum IL-6 was detected by ELISA, the expression and distribution of IL-6 in colon were observed by histochemical staining, and the effect of IL-6 on L-type calcium channel in colon smooth muscle cells was observed by whole cell patch clamp technique. The results showed that, compared with the control group, AP group exhibited reduced contractile amplitude and longer contraction cycle of colon smooth muscle strips. IL-6 prolonged the contraction cycle of colon smooth muscle strips, but did not affect their spontaneous contraction amplitude. Serum IL-6 concentration in AP group was significantly higher than that in control group (P > 0.05). IL-6 was diffusely distributed in the colon of the control group, but the expression of IL-6 was significantly up-regulated in the colon gland, mucosa and submucosa of the AP group. IL-6 significantly decreased the peak current density of L-type calcium channel in rat colon smooth muscle cells. These results suggest that the colon motility of AP rats is weakened, and the mechanism may be that up-regulated IL-6 inactivates L-type voltage-dependent calcium channels, and then inhibits the contraction of colon longitudinal smooth muscle.
Animals ; Calcium Channels, L-Type ; metabolism ; Colon ; Interleukin-6 ; metabolism ; Muscle Contraction ; Muscle, Smooth ; physiopathology ; Pancreatitis ; physiopathology ; Rats

OBJECTIVE: To investigate the anti-inflammatory effect and mechanisms of interleukin-35 (IL-35) in inflammatory bowel disease. METHODS: BALB/c mice were divided into three groups with 10 mice in each group:control group, model group (oral administration of 4% glucan sodium sulfate for 7 d) and IL-35-treated group (oral administration of 4% glucan sodium sulfate for 7 d, intraperitoneal injection of 2 μg IL-35 at d2-5). Disease activity index (DAI) was scored every day. After 7 d, the mice were sacrificed, and the serum and intestinal tissue samples were collected. The gross morphology of the colon was observed; HE staining was used to observe the pathological changes of colon tissue; flow cytometry was employed to detect the change of macrophage polarization ratio in colon tissue; the mRNA expression levels of cytokines IL-6, TNF-α, IFN-γ, IL-10 and SHIP1 in colon tissue were determined by real-time quantitative RT-PCR; the expression and distribution of SHIP1 in colon tissue was measured by immunohistochemistry; Western blotting was adopted to detect the expression level of SHIP1 protein in colonic intestinal tissues of each group. RESULTS: The DAI scores of the mice in the model group were higher than those in the control group, while the DAI scores in the IL-35-treated group were lower than those in the model group (all <0.01). Compared with the control group, the colon length was significantly shortened in the model group (<0.05), while the colon length of the IL-35-treated group had an increasing trend compared with the model group, but the difference was not statistically significant (>0.05). Compared with the model group, microscopic inflammatory infiltration score was decreased and microscopic crypt destruction and score was significantly lower in IL-35-treated group (all <0.05). The relative expression of proinflammatory cytokines IL-6, TNF-α and IFN-γ in the colon tissue of IL-35-treated group was decreased compared with the model group, while the relative expression of IL-10 mRNA was higher than that of the model group (all <0.05). Compared with the control group, the proportion of M1 macrophages in the model group increased (<0.05), while the proportion of M1 macrophages in the IL-35-treated group was lower than that in the model group (<0.05). The relative expression of SHIP1 mRNA and protein in the colon tissue of IL-35-treated group was higher than that in the model group (all <0.05). CONCLUSIONS IL-35 can inhibit the polarization of M1 macrophages and regulate inflammatory cytokines to promote anti-inflammatory effect on mice with colitis.
Animals ; Anti-Inflammatory Agents ; pharmacology ; Colitis ; drug therapy ; physiopathology ; Colon ; drug effects ; Cytokines ; genetics ; Disease Models, Animal ; Gene Expression Regulation ; drug effects ; Glucans ; pharmacology ; Interleukin-6 ; genetics ; Interleukins ; pharmacology ; Macrophages ; drug effects ; Mice ; Mice, Inbred BALB C ; Phosphatidylinositol-3,4,5-Trisphosphate 5-Phosphatases ; genetics

OBJECTIVETo establishment and verify pelvic nerve denervation (PND) model in mice.

METHODS(1) Establishment of models. Seventy-two healthy male SPE class C57 mice with age of 7 weeks and body weight of (25±1) g were chosen. These 72 mice were randomly divided into PND group containing 36 mice and sham operation group containing 36 mice. Referring to the establishment method of PND rats, after anesthesia, a laparotomy was performed on the mouse with an abdominal median incision. Under the dissection microscope, the pelvic nerves behind and after each sides of the prostate gland were bluntly separated with cotton swabs and cut with a dissecting scissor. After the operation, the urination of mice was assisted twice every day. For the mice of sham operation group, the pelvic nerves were only exposed without cutting. (2) Detection of models. Colonic transit test was performed in 18 mice chosen randomly from each group to detect the colonic transit ratio (colored colon by methylene blue/ whole colon) and visceral sensitivity tests was performed in the rest mice to observe and record the changes of electromyogram.

RESULTSThree mice died of colonic transit test in each group. Uroschesis occurred in all the mice of PND group and needed bladder massage to assist the urination. Colonic transit test showed that the colonic transit ratios of sham operation group at postoperative day (POD) 1, 3 and 7 were (0.4950±0.3858)%, (0.6386±0.1293)% and (0.6470±0.1088)% without significant difference (F=0.3647, P=0.058), while in PND group, the colonic transit ratio at POD 7 [(0.6044±0.1768) %] was obviously higher than that both at POD 3[(0.3876±0.1364)%, P=0.022] and POD 1[(0.2542±0.0371)%, P=0.001], indicating a recovery trend of colonic transit function (F=9.143, P=0.004). Compared with the sham operation group, the colonic transit function in PND group decreased significantly at POD 1 and POD 3(both P<0.05), and at POD 7, there was no significant difference between two groups. Visceral sensitivity test showed that the visceral sensitivity of sham operation group at POD 1, 3 and 7 was 24.2808±9.5566, 33.6725±7.9548 and 43.9086±12.1875 with significant difference (F=5.722, P=0.014). The visceral sensitivity of PND group at POD 1, 3 and 7 was 11.7609±2.1049, 21.8415±8.1527 and 26.2310±4.2235 with significant difference as well (F=11.154, P=0.001). The visceral sensitivity at POD 3 and POD 7 was obviously higher than that at POD 1 (P=0.006, P<0.001), and there was no significant difference between POD 3 and POD 7 (P=0.183). Compared with sham operation group, the visceral sensitivity of PND group decreased significantly at POD 1, 3 and 7(all P<0.05).

CONCLUSIONSDenervation of pelvic nerves can obviously decrease the colonic transit function and the visceral sensitivity of mice, but these changes can recover over time, which suggests that the establishment of PND model in mice is successful.

Abdominal Pain ; physiopathology ; Animals ; Autonomic Pathways ; growth & development ; physiopathology ; surgery ; Colon ; innervation ; physiopathology ; Denervation ; methods ; Disease Models, Animal ; Gastrointestinal Transit ; physiology ; Male ; Mice ; Mice, Inbred C57BL ; Nerve Tissue ; growth & development ; physiopathology ; surgery ; Pain, Postoperative ; physiopathology ; Pelvis ; innervation ; physiopathology ; surgery ; Prostate ; innervation ; Recovery of Function ; physiology

OBJECTIVETo investigate the risk factors of low anterior resection syndrome (LARS) after anal sphincter preserving surgery (SPS) for rectal cancer patients.

METHODSClinicopathological and follow-up data of rectal cancer patients who underwent SPS from January 2010 to June 2014 in Department of Gastroenterological Surgery, Peking University People's Hospital, were retrospectively analyzed. Patients receiving permanent colostomy and local resection were excluded. Meanwhile, during October 2014 and March 2015, the enrolled patients were asked to fill out a specially designed questionnaire for LARS through face-to-face interview or telephone inquiry, according to the chronological order of operation. Based on the score of questionnaire, patients were divided into three groups: 0-20 points: non LARS; 21-29: minor LARS; 30-42: major LARS. The demographic and clinicopathologic features were compared among groups and the risk factors of major LARS were tested by logistic regression analysis.

RESULTSA total of 100 patients (61 males, 39 females) completed the bowel function survey, with an average age of 66.2(41-86) years, 33 patients <60 years versus 67 patients ≥60 years. No significant difference was observed in age distribution (P=0.204). Interval from operation to first follow-up was more than 1 year in 70 patients, and the median follow-up was 23 months. Thirty-seven patients were non LARS, 18 were minor LARS and 45 were major LARS. No significant differences in clinicopathological data (all P>0.05) were observed among three groups except radiotherapy history (P=0.025), tumor location(P=0.000) and distance from anastomotic site to anal verge(P=0.008). After comparison of non LARS group combined with minor LARS group versus major LARS, re-analysis of risk factors showed that radiotherapy history (RR=5.608, 95%CI:1.457 to 21.584, P=0.006), distance from tumor lower margin to anal verge (RR=0.125, 95%CI:0.042 to 0.372, P=0.000), distance from anastomotic site to anal verge (RR=0.255, 95%CI:0.098 to 0.665, P=0.004) and preventive ileostomy history(RR=3.643, 95%CI:1.058 to 12.548, P=0.032) were associated with major LARS. One potential risk factor detected in combined analysis was female (RR=2.138, 95%CI: 0.944 to 4.844, P=0.078). Multivariate analysis revealed that female (RR=2.654, 95%CI: 1.005 to 7.014, P=0.049), radiotherapy history (RR=10.422, 95%CI:2.394 to 45.368, P=0.002) and distance from tumor lower margin to anal verge ≤7 cm (RR=8.935, 95%CI:2.827 to 28.243, P=0.000) were independent risk factors of major LARS.

CONCLUSIONSLARS is a significant problem in most rectal cancer patients after SPS. The risk of major LARS increases on condition of radiotherapy, low tumor position and female. When dealing with these patients, preventive measures should be taken into consideration during SPS.

Adult ; Aged ; Aged, 80 and over ; Anal Canal ; pathology ; surgery ; Anastomosis, Surgical ; adverse effects ; Colon ; surgery ; Defecation ; physiology ; Factor Analysis, Statistical ; Female ; Follow-Up Studies ; Humans ; Long Term Adverse Effects ; epidemiology ; Male ; Middle Aged ; Radiotherapy ; adverse effects ; Rectal Neoplasms ; complications ; pathology ; surgery ; Rectum ; physiopathology ; surgery ; Retrospective Studies ; Risk Assessment ; methods ; Risk Factors ; Sex Factors ; Surveys and Questionnaires ; Syndrome

OBJECTIVETo investigated the involvement of pulmonary function impairment in ulcerative colitis (UC), to explore a scientific basis for the Chinese medicine (CM) theory of exterior-interior correlation between Lung (Fei) and Large intestine (Dachang).

METHODSTotally 120 patients with a diagnosis of UC were recruited and the demographics, clinical data, and blood samples were collected. C-reactive protein (CRP) and erythrocyte sedimentation rate (ESR) concentrations were measured. Every patient accepted pulmonary function test and took chest radiograph (CXR).> RESULTS: Pulmonary function abnormalities were present in 72 of 120 patients. The median (interquartile range) vital capacity (VC), forced vital capacity (FVC), forced expiratory volume in 1 s (FEV), carbon monoxide diffusion capacity (DL) of lung, total lung capacity (TLC) and functional residual volume (FRV) were decreased in distal UC and pancolitis compared with ulcerative prochitis (P <0.0005). Male patients had increased VC, FEV/FVC, and residual volume (RV)/TLC compared with female (P <0.0005), but decreased DLand carbon monoxide iffusion capacity (K) of lung/alveolar ventilation (P <0.0005). Age was strongly correlated with RV (Spearman rank correlation coefficient (rs)=-0.57,P <0.0001), and RV/TLC (rs=0.48,P<0.0001). Age was also correlated with FEV/FVC (rs=-0.29, P=0.001), forced expiratory flow in 75% vital capacity (FEF75%, rs=-0.20, P=0.03), DL(rs=-0.21, P=0.02), TLC (rs=-0.25, P=0.006), and FRV (rs=-0.28, P=0.002). The course of disease was correlated with FEF75% (rs=-0.18, P=0.049) and K(rs=-0.19, P=0.036). Chest radiograph abnormalities were presented in 38 of 120. Pulmonary symptoms were presented in 10 of 120. Other extraintestinal complications were presented in 21 of 120.

CONCLUSIONSPulmonary function impairment was more frequently than other extraintestinal complications in UC patients, which may be affected by sex, age, extent and course of disease. These results may be a scientific basis for the theory of exterior-interior correlation between Lung and Large intestine.

Adolescent ; Adult ; Age of Onset ; Aged ; Colitis, Ulcerative ; complications ; pathology ; physiopathology ; Colon ; pathology ; Demography ; Female ; Humans ; Inflammation ; complications ; pathology ; Lung ; diagnostic imaging ; physiopathology ; Male ; Middle Aged ; Radiography, Thoracic ; Respiratory Function Tests ; Young Adult

OBJECTIVETo study the effects of sacral nerve root electrostimulation (SNS) on the colon function and its mechanisms in rats with spinal cord injury (SCI).

METHODSOne hundred and four Wistar rats were divided into three groups: A, B and C. A group ( n = 24) was divided into three subgroups (n = 8) for studying the bioelectricity: Normal group (NG), SCI group (SCI) and SCI group with SNS(SNS); B group( n = 24) was divided into three subgroups( n = 8) for studying the colon motility: NG, SCI and SNS. C group( n = 56) were divided into three groups for studying the change of morphology and neurotransmitters(SP and VIP): NG (n = 8), SCI (n = 24), and SNS (n = 24) . In SCI and SNS, included of three subgroups: 24, 48, 72 h after spinal cord injury (n = 8).

RESULTSIn SCI group, the activity of bioelectricity in proximal and distal colon was reduced; the colon motility was lessened, and colon mucosa appeared different degree of damage; cell-cell connections between intestinal epithelial cells were destroyed. The expressions of substance P(SP) and vasoactive intestinal peptide (VIP) in colon were decreased obviously. SNS was found to activate the bioelectricity, promote the colon motility, improve the intestinal mucosal, and increase the expressions of SP and VIP. Conclusion: SNS can activate the peristalsis, rehabilitate the motility of denervated colon, protection of the intestinal mechanical barrier between intestinal epithelial cells and tight junction, rebuild the colon function through activating the bioelectricity and increase the expressions of SP and VIP.

Animals ; Colon ; physiopathology ; Electric Stimulation Therapy ; Epithelial Cells ; drug effects ; Intestinal Mucosa ; drug effects ; Lumbosacral Region ; innervation ; Neurotransmitter Agents ; metabolism ; Rats ; Rats, Wistar ; Spinal Cord Injuries ; therapy ; Substance P ; metabolism ; Vasoactive Intestinal Peptide ; metabolism

OBJECTIVETo investigate the application value of colonic transit test (CTT) combined with anorectal manometry (ARM), barium enema (BE) and defecography (DFG) in accurately evaluating colonic lesions of slow transit constipation complicated with adult megacolon.

METHODSClinical data of 47 above patients admitted between October 2007 and February 2015 in the People's Hospital of Hunan Province were analyzed retrospectively. All the patients were examined with≥2 times of CTT combined with ARM and BE, and 42 cases received additional DFG at the same time, to evaluate colonic lesions before operation. Operative biopsy pathology was used as the standard. The sensitivity, specificity, positive predictive value(PPV) and negative predictive value(NPV) of positioning in the ascending colon and caecum, transverse colon and descending colon were calculated, and the consistency was represented by Kappa test(Kappa value≥0.75 indicates good consistency, meanwhile higher Kappa value indicates better consistency). The Heikkinen score was used to evaluate defecation function at postoperative 6 months.

RESULTSThe age of 47 patients was from 18 to 56 years old. Compared with intraoperative findings and biopsy pathology, the diagnostic coincidence rate was 89.4% by CTT combined with BE and DFG positioning, which suggested pathology-changed colonic segment locating in the ascending colon and cecum (n=12), transverse colon (n=26) and descending colon (n=9), while intraoperative findings and biopsy pathology suggested pathology-changed colonic segment locating in the ascending colon and cecum (n=11), transverse colon (n=23) and descending colon (n=13). The sensitivity was 88.3%, specificity 93.5%, PPV 92.1%, NPV 94.9% and Kappa value was 0.827(P<0.001). Procedures performed included segmental colectomy (n=8), subtotal colectomy (n=29), total colectomy (n=10). There was no serious complication during and after operation. Defecatory function was excellent in 24 cases (60.0%), good in 10 (25.0%), and moderate in 6 (15.0%) evaluated by Heikkinen score at postoperative 6 months. A total of 40 patients were followed up from 1 to 7 years (median 3 years) and there was no long-term diarrhea and recurrence of constipation or giant colon after operation.

CONCLUSIONPreoperative detection of CTT combined with ARM, BE and DFG in patients with slow transit constipation complicated with adult megacolon can make a more precise assessment of the extent of colonic lesions in advance, which has a good clinical predictive value.

Adolescent ; Adult ; Barium Enema ; Cecum ; pathology ; physiopathology ; surgery ; Colectomy ; methods ; Colon ; pathology ; physiopathology ; surgery ; Constipation ; complications ; diagnosis ; pathology ; surgery ; Defecography ; Female ; Gastrointestinal Transit ; physiology ; Humans ; Male ; Manometry ; Megacolon ; complications ; pathology ; surgery ; Middle Aged ; Predictive Value of Tests ; Preoperative Period ; Prognosis ; Recovery of Function ; physiology ; Retrospective Studies ; Sensitivity and Specificity

To observe the effect of vasoactive intestinal peptide (VIP) on the metabolism of intestinal fluid and cyclic AMP protein kinase A signaling pathway (cAMP-PKA) and water channel protein 3 (AQP3) in rats with constipation, and to explore the mechanism of VIP in the treatment of constipation.
 Methods: A total of 45 healthy adult rats were randomly divided into a control group, a model group, a model +VIP group. After 4 weeks of VIP treatment, the first black stool time were examined with the ink gastric method; the water content in feces was calculated; the morphological changes in colonic tissues were observed by HE staining. The expression of VIP and AQP3 protein levels in colon tissues were detected by Western blot; and the cAMP, PKA, AQP3 mRNA expression levels were detected by quantitative real time polymerase chain reaction (qPCR). 
 Results: Compared with the control group, the first black stool time was prolonged, the water content of fecal decreased significantly (both P<0.01); part of the colon mucosa epithelial cells were destructed; the goblet cell volume decreased and quantity was reduced; the contents of AQP3 and VIP in colon tissues were significantly decreased, and the cAMP, PKA and AQP3 mRNA levels were decreased in the model group (all P<0.05). Compared with the model group, the first black stool time in the model +VIP group was shortened, the fecal water content increased significantly (both P<0.05); the mucosal epithelium integrity improved, the number of goblet cells increased; the content of AQP3 and VIP in colon tissues was increased, and the cAMP, PKA, and AQP3 mRNA levels were elevated (all P<0.05).
 Conclusion: Intravenous injection of VIP can regulate intestinal fluid metabolism and improve the symptoms of constipation in rats, which might be related to the regulation of VIP-cAMP-PKA-AQP3 signaling pathway.
Animals ; Aquaporin 3 ; physiology ; Aquaporins ; Blotting, Western ; Colon ; chemistry ; pathology ; Constipation ; physiopathology ; therapy ; Cyclic AMP ; physiology ; Defecation ; Epithelial Cells ; pathology ; Feces ; chemistry ; Goblet Cells ; pathology ; Intestinal Mucosa ; metabolism ; pathology ; RNA, Messenger ; Rats ; Signal Transduction ; Vasoactive Intestinal Peptide ; administration & dosage ; physiology ; therapeutic use

BACKGROUND/AIMS: This animal study aimed to define the underlying cellular mechanisms of intestinal barrier dysfunction. METHODS: Rats were fed 4% with dextran sodium sulfate (DSS) to induce experimental colitis. We analyzed the sugars in 24-hour urine output by high pressure liquid chromatography. The expression of claudins, mannan-binding lectin (MBL), and MBL-associated serine proteases 2 (MASP-2) were detected in the colonic mucosa by immunohistochemistry; and apoptotic cells in the colonic epithelium were detected by the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling method assay. RESULTS: The lactulose and sucralose excretion levels in the urine of rats with DSS-induced colitis were significantly higher than those in the control rats. Mannitol excretion was lower and lactulose/mannitol ratios and sucralose/mannitol ratios were significantly increased compared with those in the control group (p<0.05). Compared with the controls, the expression of sealing claudins (claudin 3, claudin 5, and claudin 8) was significantly decreased, but that of claudin 1 was increased. The expression of pore-forming claudin 2 was upregulated and claudin 7 was downregulated in DSS-induced colitis. The epithelial apoptotic ratio was 2.8%+/-1.2% in controls and was significantly increased to 7.2%+/-1.2% in DSS-induced colitis. The expression of MBL and MASP-2 in the intestinal mucosa showed intense staining in controls, whereas there was weak staining in the rats with colitis. CONCLUSIONS: There was increased intestinal permeability in DSS-induced colitis. Changes in the expression and distribution of claudins, increased epithelial apoptosis, and the MASP-2-induced immune response impaired the intestinal epithelium and contributed to high intestinal permeability.
Animals ; Apoptosis/*physiology ; Claudins/*metabolism ; Colitis/chemically induced/immunology/*physiopathology ; Colon/immunology/physiopathology ; Dextran Sulfate ; Intestinal Mucosa/*physiopathology ; Lactulose/metabolism ; Mannitol/metabolism ; Mannose-Binding Lectin/*immunology ; Permeability ; Rats ; Rats, Sprague-Dawley ; Sucrose/analogs & derivatives/metabolism ; Up-Regulation

OBJECTIVETo compare the effects of electroacupuncture (EA) and moxibustion therapies on patients with diarrhea-predominant irritable bowel syndrome (D-IBS).

METHODSA total of 60 D-IBS patients were randomly allocated to the EA group (30 cases) and moxibustion group (30 cases). Before and after treatment, the gastrointestinal symptoms and psychological symptoms were scored by Visual Analogue Scale, Bristol Stool Form Scale, Hamilton Anxiety Rating Scale (HAMA), and Hamilton Depression Rating Scale (HAMD); the expressions of 5-hydroxytryptamine (5-HT), 5-HT3 receptor (5-HT3R), and 5-HT4 receptor (5-HT4R) in the sigmoid mucosal tissue were measured by immunohistochemical staining. Additionally, the effects on the functional brain areas of the anterior cingulate cortex (ACC), insular cortex (IC) and prefrontal cortex (PFC) were observed by functional magnetic resonance imaging.

RESULTSCompared with before treatment, both EA and moxibustion groups reported significant improvements in abdominal pain and abdominal bloating after treatment (P<0.01 or P<0.05). The moxibustion group reported greater improvements in defecation emergency, defecation frequency, and stool feature than the EA group (P<0.01). Both HAMA and HAMD scores were significantly decreased in the moxibustion group than in the EA group (P<0.01). Both groups demonstrated significantly reduced expressions of 5-HT, 5-HT3R and 5-HT4R in the colonic mucosa after treatment (P<0.01), with a greater reduction of 5-HT in the moxibustion group (P<0.05). Finally, decreased activated voxel values were observed in the left IC, right IC and PFC brain regions of patients in the moxibustion group under stimulation with 150 mL colorectal distension after treatment (P<0.05 or P<0.01), while in the EA group only PFC area demonstrated a reduction (P<0.05).

CONCLUSIONMoxibustion can significantly improve the symptoms of D-IBS, suggesting that moxibustion may be a more effective therapy than EA for D-IBS patients.

Adult ; Anxiety ; Brain ; physiology ; Cerebral Cortex ; physiopathology ; Colon, Sigmoid ; chemistry ; Depression ; Diarrhea ; physiopathology ; Electroacupuncture ; Gastrointestinal Tract ; physiology ; Gyrus Cinguli ; physiopathology ; Humans ; Immunohistochemistry ; Intestinal Mucosa ; chemistry ; Irritable Bowel Syndrome ; physiopathology ; psychology ; therapy ; Magnetic Resonance Imaging ; Moxibustion ; Pain Measurement ; Prefrontal Cortex ; physiopathology ; Receptors, Serotonin, 5-HT3 ; analysis ; Serotonin ; analysis