Animals ; Antibodies, Neutralizing/biosynthesis* ; Antibodies, Viral/biosynthesis* ; Body Weight/immunology* ; COVID-19/virology* ; Disease Models, Animal ; Disease Progression ; Humans ; Immunohistochemistry ; Lung/virology* ; Male ; Mesocricetus ; Nasal Cavity/virology* ; RNA, Viral/immunology* ; SARS-CoV-2/pathogenicity* ; Severity of Illness Index ; Viral Load

OBJECTIVETo investigate the safety, efficacy and long-term outcomes of Jinling procedure in the treatment of adult Hirschsprung disease.

METHODSClinical and follow-up data of 125 patients with adult Hirschsprung disease undergoing Jinling procedure at the Department of General Surgery between January 2000 and January 2013 were summarized. All the patients were diagnosed by CT, barium enema, anorectal pressure detection and pathology examination. Abdominal symptoms, gastrointestinal quality of life index(GIQLI, the lower score, the worse quality of life), Wexner constipation score (higher score indicated worse symptom), defecography (evaluation included rectocele, mucosal prolapse, intramucosal intussusception, perineal prolapse) and other operative complications were compared before and after operation.

RESULTSAmong 125 patients, 69 were male and 56 were female with median age of (41.2±15.5) (18 to 75) years. The follow-up rates were 94.4%(118/125), 92.0%(115/125), 89.6%(112/125) and 88.0%(110/125) at postoperative months 1, 3, 6, and 12. Incidences of abdominal distension and abdominal pain were 100% and 82.4%(103/125) before operation, and were 7.3%(8/110) and 20.9%(23/110) at 12 months after surgery. Wexner score was significantly lower at postoperative months 1(8.7±2.9), 3 (7.2±2.8), 6(6.7±2.2) and 12(6.3±1.7) than that before operation (21.4±7.2) (P<0.01). GIQLI score was 51.6±11.9 before operation, though it decreased at 1 month (47.3±5.5)(P<0.05) after surgery, but increased significantly at postoperative months 3, 6, 12(68.9±8.0, 96.5±8.2, 103.2±8.6)(P<0.01). Abnormal rate of defecography was 70.4%(81/115), 48.2%(54/112) and 27.3%(30/110) at postoperative months 3, 6, 12, which was significantly lower than 91.2%(114/125) before operation (P<0.01). Morbidity of postoperative complication was 29.6%(37/125), including 5 cases of surgical site infection (4.0%), 2 of anastomotic bleeding (1.6%), 8 of anastomotic leakage (6.4%, one died of severe abdominal infection), 4 of urinary retention (3.2%), 3 of recurrent constipation (2.4%, without megacolon relapse), 11 of bowel obstruction (8.8%), 2 of anastomotic stricture(1.6%) and 2 of refractory staphylococcus aureus enteritis (1.6%, diagnosed by stool smear and culture, and both died finally).

CONCLUSIONJinling procedure is a safe and effective surgical procedure for adult Hirschsprung's disease.

Adolescent ; Adult ; Aged ; Anastomosis, Surgical ; Colectomy ; Constipation ; Defecography ; Digestive System Surgical Procedures ; Female ; Hirschsprung Disease ; surgery ; Humans ; Intestinal Obstruction ; Intussusception ; Male ; Middle Aged ; Perineum ; Postoperative Complications ; Postoperative Period ; Quality of Life ; Rectocele ; Staphylococcus aureus ; Treatment Outcome ; Young Adult

OBJECTIVETo investigate the expression of TMEMl6A in the colon of intractable functional constipation patients and its clinical implications.

METHODSA total of 30 patients with intractable chronic functional constipation were selected as trial group (full thickness tissue of sigmoid colon), 30 patients with colon cancer as control group (distant tissues at least 5 cm away from cancer). Tissues from two groups were collected in our hospital from February 2012 to June 2014 and confirmed by pathological diagnosis. Immunofluorescence staining, RT-PCR and Western blot were performed to detect the mRNA and protein expression of TMEM16A and c-kit in colon.

RESULTSTMEM16A and c-kit protein expressions were observed in similar sites of colon tissues in two groups. Expressions of TMEM16A and C-kit in colon tissues detected by immunofluorescence, RT-PCR, and Western blotting were significantly lower in trial group than those in control group (TMEM16A: mean A 1.84±0.25 vs. 3.65±0.32, P<0.05, gray intensity ratio 0.66±0.07 vs. 1.04±0.17, P<0.05, relative mRNA 0.41±0.05 vs. 1.00, P<0.05; c-kit: mean A 3.38±0.24 vs. 5.06±0.31, gray intensity ratio 0.64±0.06 vs. 0.98±0.09, relative mRNA 0.18±0.08 vs. 1.00, all P<0.05).

CONCLUSIONSTMEM16A expression in colon tissues of intractable functional constipation patients is significantly lower and may adjust the movement of colonic smooth muscle by regulating the c-kit expression. TMEMl6A may be used as a new candidate target for diagnosis and treatment of intractable functional constipation.

Anoctamin-1 ; Blotting, Western ; Chloride Channels ; Colon, Sigmoid ; Colonic Neoplasms ; Constipation ; Humans ; Muscle, Smooth ; Neoplasm Proteins ; Proteomics ; Proto-Oncogene Proteins c-kit ; RNA, Messenger

OBJECTIVETo establish an innovative rat model of slow transit constipation by selective chemical ablation of the colon enteric plexus.

METHODSSprague Dawley rats, 5-6 weeks old, were randomly divided into normal control group, sham operation group, treatment group I, II, III, IIII. The normal control group did not receive treatment. Rats in the sham operation group and the treatment groups received abdominal operation under anesthesia, and the gauze containing 0.9% normal saline, 0.05%, 0.1%, 0.25%, 0.5% benzalkonium chloride (BAC) was applied into colon for 30 minutes. Two weeks after operation, the number of feces, fecal dry weight in 24 h and gastrointestinal transit time were recorded, then hematoxylin-eosin (HE) staining, immunohistochemistry, ELISA were used for the evaluation of colonic pathology, enteric plexus, Interstitial cells of Cajal and neurotransmitters 5-hydroxytryptamine(5-HT).

RESULTSCompared to the normal control group and the sham operation group, the gastrointestinal transit time was significantly prolonged and fecal dry weight was lower in the treatment group II, III (all P<0.05). HE and immunohistochemical staining showed varying degrees of pathological changes in the treatment groups and in line with the pathological changes of slow transit constipation. 5-HT concentration reduced significantly in treatment group III compared to other groups (P<0.01).

CONCLUSIONThe animal model of STC is successfully established by applying 0.25% BAC selective chemical ablation of the colon enteric plexus. This model is simple, stable, and is more in line with pathological changes of slow transit constipation.

Animals ; Autonomic Pathways ; Constipation ; Disease Models, Animal ; Gastrointestinal Transit ; Immunohistochemistry ; Interstitial Cells of Cajal ; Rats ; Rats, Sprague-Dawley

Objective To establish an innovative rat model of slow transit constipation by selective chemical ablation of the colon enteric plexus. Methods Sprague Dawley rats, 5-6 weeks old, were randomly divided into normal control group, sham operation group, treatment groupⅠ, Ⅱ, Ⅲ,Ⅳ. The normal control group did not receive treatment. Rats in the sham operation group and the treatment groups received abdominal operation under anesthesia , and the gauze containing 0.9% normal saline, 0.05%, 0.1%, 0.25%, 0.5% benzalkonium chloride (BAC) was applied into colon for 30 minutes. Two weeks after operation, the number of feces, fecal dry weight in 24 h and gastrointestinal transit time were recorded, then hematoxylin-eosin (HE) staining, immunohistochemistry, ELISA were used for the evaluation of colonic pathology, enteric plexus, Interstitial cells of Cajal and neurotransmitters 5-hydroxytryptamine (5-HT). Results Compared to the normal control group and the sham operation group, the gastrointestinal transit time was significantly prolonged and fecal dry weight was lower in the treatment groupⅡ, Ⅲ (all P<0.05). HE and immunohistochemical staining showed varying degrees of pathological changes in the treatment groups and in line with the pathological changes of slow transit constipation. 5-HT concentration reduced significantly in treatment group Ⅲ compared to other groups (P<0.01). Conclusion The animal model of STC is successfully established by applying 0.25% BAC selective chemical ablation of the colon enteric plexus. This model is simple , stable, and is more in line with pathological changes of slow transit constipation.

Objective To investigate the expression of TMEMl6A in the colon of intractable functional constipation patients and its clinical implications. Methods A total of 30 patients with intractable chronic functional constipation were selected as trial group (full thickness tissue of sigmoid colon), 30 patients with colon cancer as control group (distant tissues at least 5 cm away from cancer). Tissues from two groups were collected in our hospital from February 2012 to June 2014 and confirmed by pathological diagnosis. Immunofluorescence staining, RT-PCR and Western blot were performed to detect the mRNA and protein expression of TMEM16A and c-kit in colon. Results TMEM16A and c-kit protein expressions were observed in similar sites of colon tissues in two groups. Expressions of TMEM16A and C-kit in colon tissues detected by immunofluorescence, RT-PCR, and Western blotting were significantly lower in trial group than those in control group (TMEM16A: mean A 1.84±0.25 vs. 3.65±0.32, P<0.05, gray intensity ratio 0.66±0.07 vs. 1.04±0.17, P<0.05, relative mRNA 0.41±0.05 vs. 1.00, P<0.05﹔ c-kit: mean A 3.38±0.24 vs. 5.06±0.31, gray intensity ratio 0.64±0.06 vs. 0.98± 0.09, relative mRNA 0.18 ±0.08 vs. 1.00, all P<0.05). Conclusions TMEM16A expression in colon tissues of intractable functional constipation patients is significantly lower and may adjust the movement of colonic smooth muscle by regulating the c-kit expression. TMEMl6A may be used as a new candidate target for diagnosis and treatment of intractable functional constipation.

Objective To establish an innovative rat model of slow transit constipation by selective chemical ablation of the colon enteric plexus. Methods Sprague Dawley rats, 5-6 weeks old, were randomly divided into normal control group, sham operation group, treatment groupⅠ, Ⅱ, Ⅲ,Ⅳ. The normal control group did not receive treatment. Rats in the sham operation group and the treatment groups received abdominal operation under anesthesia , and the gauze containing 0.9% normal saline, 0.05%, 0.1%, 0.25%, 0.5% benzalkonium chloride (BAC) was applied into colon for 30 minutes. Two weeks after operation, the number of feces, fecal dry weight in 24 h and gastrointestinal transit time were recorded, then hematoxylin-eosin (HE) staining, immunohistochemistry, ELISA were used for the evaluation of colonic pathology, enteric plexus, Interstitial cells of Cajal and neurotransmitters 5-hydroxytryptamine (5-HT). Results Compared to the normal control group and the sham operation group, the gastrointestinal transit time was significantly prolonged and fecal dry weight was lower in the treatment groupⅡ, Ⅲ (all P<0.05). HE and immunohistochemical staining showed varying degrees of pathological changes in the treatment groups and in line with the pathological changes of slow transit constipation. 5-HT concentration reduced significantly in treatment group Ⅲ compared to other groups (P<0.01). Conclusion The animal model of STC is successfully established by applying 0.25% BAC selective chemical ablation of the colon enteric plexus. This model is simple , stable, and is more in line with pathological changes of slow transit constipation.

Objective To investigate the expression of TMEMl6A in the colon of intractable functional constipation patients and its clinical implications. Methods A total of 30 patients with intractable chronic functional constipation were selected as trial group (full thickness tissue of sigmoid colon), 30 patients with colon cancer as control group (distant tissues at least 5 cm away from cancer). Tissues from two groups were collected in our hospital from February 2012 to June 2014 and confirmed by pathological diagnosis. Immunofluorescence staining, RT-PCR and Western blot were performed to detect the mRNA and protein expression of TMEM16A and c-kit in colon. Results TMEM16A and c-kit protein expressions were observed in similar sites of colon tissues in two groups. Expressions of TMEM16A and C-kit in colon tissues detected by immunofluorescence, RT-PCR, and Western blotting were significantly lower in trial group than those in control group (TMEM16A: mean A 1.84±0.25 vs. 3.65±0.32, P<0.05, gray intensity ratio 0.66±0.07 vs. 1.04±0.17, P<0.05, relative mRNA 0.41±0.05 vs. 1.00, P<0.05﹔ c-kit: mean A 3.38±0.24 vs. 5.06±0.31, gray intensity ratio 0.64±0.06 vs. 0.98± 0.09, relative mRNA 0.18 ±0.08 vs. 1.00, all P<0.05). Conclusions TMEM16A expression in colon tissues of intractable functional constipation patients is significantly lower and may adjust the movement of colonic smooth muscle by regulating the c-kit expression. TMEMl6A may be used as a new candidate target for diagnosis and treatment of intractable functional constipation.