Objective To investigate the effects of pelvic nerve transection on the colonic motility and the expression of transit receptor potential ankyrin 1 (TRPA1) in the colon mucosa.Methods Ninety-six Sprague-Dawley rats were divided into 3 groups based on a random number table:(1) 18 rats in the control group remained untreated and were fed regularly;(2) 39 rats in the sham operation group received laparotomy,and the pelvic nerves were stripped;(3) 39 rats in the operation group received laparotomy with pelvic nerve transection before abdominal closure.Colonic transit was assessed respectively at postoperative day 1,3,and 7 by injecting and calculating the geometric center (GC) of the distribution of 51Cr after 3 hours of propagation.The expression of TRPA1 in the colonic mucosa was determined by Western blot at postoperative day 1,3,7.Data with normal distribution were expressed by (x)± s,and were analyzed using the repeated measures ANOVA or LSD test.Results The GC values of the distribution of 51Cr in the sham operation group at postoperative day 1,3,7 were 5.8 ± 0.9,7.5 ± 0.5,7.3 ± 0.5,with a significant difference (F =9.508,P ＜ 0.05).The GC values of the distribution of 51Cr in the operation group at postoperative day 1,3,7 were 4.9 ± 0.4,5.6 ± 0.4,6.4 ± 0.8,with a significant difference (F =11.689,P ＜ 0.05).There were significant differences in the GC values of the distribution of 51 Cr at postoperative day 1 and 3 between the sham operation group and the operation group (t =2.227,7.144,P ＜ 0.05),while no significant difference was detected at postoperative day 7 (t =2.162,P ＞ 0.05).The results of Western blot showed that the relative expressions of TRPA1 in the proximal part of the colon at postoperative day 1,3,7 were 1.00 ± 0.05,1.00 ± 0.07,1.00 ± 0.06 in the control group,with a significant difference (F =0.055,P ＞ 0.05).The relative expressions of TRPA1 in the proximal part of the colon at postoperative day 1,3,7 were 0.78 ± 0.09,0.94 ± 0.08,0.95 ± 0.12 in the sham operation group,with a significant difference (F =5.651,P ＜ 0.05).The relative expressions of TRPA1 in the proximal part of the colon at postoperative day 1,3,7 were 0.37 ± 0.12,0.89 ± 0.10,0.92 ± 0.14 in the operation group,with a significant difference (F =41.005,P ＜0.05).There was significant difference in the relative expressions of TRPA1 in the proximal part of the colon among the 3 groups at postoperative day 1 (F =73.497,P ＜ 0.05),while significant differences were respectively detected between the control group and the sham operation group and the operation group at postoperative day 1 (t =4.224,11.954,P ＜ 0.05),and significant difference between the operation group and the sham operation group was also observed (t =7.730,P ＜ 0.05).There was no significant difference in the relative expression of TRPA1 in the proximal part of the colon among the 3 groups between day 3 and day 7 (F =2.087,0.656,P ＞ 0.05).The relative expressions of TRPA1 in the distal part of the colon at postoperative day 1,3,7 were 1.00 ± 0.05,1.00 ± 0.07,1.00 ± 0.06 in the control group,with no significant difference (F =0.055,P ＞ 0.05).The relative expressions of TRPA1 in the distal part of the colon at postoperative day 1,3,7were 0.68 ±0.11,0.98 ±0.12,1.11 ±0.16 in the sham operation group,with a significant difference (F =16.975,P ＜ 0.05).The relative expressions of TRPA1 in the distal part of the colon at postoperative day 1,3,7 were 0.39 ± 0.12,0.78 ± 0.10,1.06 ± 0.13 in the operation group,with a significant difference (F =50.417,P ＜ 0.05).There were significant differences in the relative expression of TRPA1 in the distal part of the colon among the 3 groups between day 1 and day 3 (F =58.773,8.680,P ＜ 0.05),while significant differences were respectively detected between the control group and the sham operation group and the operation group at postoperative day 1 (t =5.706,10.837,P ＜ 0.05),and significant difference was also detected between the operation group and the sham operation group (t =5.131,P ＜ 0.05).There was no significant difference in the relative expression of TRPA1 in the distal part of the colon between the control group and the sham operation group at postoperative day 3 (t =0.166,P ＞ 0.05),while significant differences were respectively detected between the control group and the operation group and between the sham operation group and the operation group at postoperative day 3 (t =3.694,3.528,P ＜ 0.05).There was no significant difference in the relative expression of the TRPA1 in the distal part of the colon between the 3 groups at postoperative day 7 (F =1.319,P ＞ 0.05).Conclusions Injury to pelvic nerves adversely affects colonic transit and expression of TRPA1 in mucosa.With a compensatory mechanism from the intestinal itself,these alterations in intestinal motility function normalize over time suggesting expression of TRPA1 in mucosa plays a crucial role in the recovery of intestinal motility function.

Objective To investigate the expression of transit receptor potential ankyrin 1 (TRPA1) and 5-hydroxytryptamine (5-HT) in the colonic mucosa of patients with slow transit constipation (STC).Methods The clinical data of 10 patients with STC and 10 patients with sigmoid cancer who were respectively admitted to the Daping Hospital of the Third Military Medical University from March 2013 to March 2014 and from December 2013 to May 2014 were retrospectively analyzed.Ten and 10 surgical specimens of descending colon from patients with STC and sigmoid cancer (tumor edge measuring in controls ＞ 10 cm) were allocated into the STC group and the control group,respectively.The expressions of TRPA1 and 5-HT in the colonic mucosa in the control group and in the 2 groups were detected by immunofluorescence and immunochemistry staining,respectively.The expression of TRPA1 in the colonic mucosa was observed by Western blot test.The measurement data with normal distribution was presented as x ± s,and the comparison between groups were analyzed using the t test.Results The results of immunofluorescence showed that TRPA1 and 5-HT were co-expressed in the colonic mucosa of controls.The results of immunochemistry staining showed that the integrated optical density (IOD) of expressions of TRPA1 and 5-HT in the STC group and in the control group were 3 619 ± 1 702,55 721 ±28 613 and 9 894 ± 3 325,12 949 ± 2 200,respectively,showing significant differences between the 2 groups (t =-5.312,4.713,P ＜ 0.05).The results of Western blot test showed that the relative expressions of TRPA1 in the STC group and in the control group were 0.8 ± 0.3 and 1.5 ± 0.5,showing a significant difference between the 2 groups (t =-2.379,P ＜ 0.05).Conclusions TRPA1 is expressed in the enterochromaffin (EC) cells of colonic mucosa.High expression of 5-HT and low expression of TRPA1 in the colonic mucosa are closely related with the pathogenesis of STC.

Objective:To explore the related factors of epilepsy secondary to aneurysmal subarachnoid hemorrhage in the elderly.Methods:From January 2015 to January 2019, 213 elderly patients with aneurysmal subarachnoid hemorrhage admitted to Wuhan Hospital of Traditional Chinese Medicine were divided into epilepsy group(46 cases) and non-epilepsy group(167 cases) according to whether secondary epilepsy.Univariate analysis was used to analyze the related factors affecting epilepsy secondary to aneurysmal subarachnoid hemorrhage in the elderly, and multivariate logistic regression was used to analyze the risk factors of epilepsy secondary to aneurysmal subarachnoid hemorrhage in the elderly.The investigation factors included sex, age, hypertension, diabetes mellitus, smoking history, location of responsible aneurysms, number of aneurysms, intracranial hematoma, hydrocephalus and neurological sequelae.Results:Univariate analysis showed that there were no statistically significant differences in sex, age, history of diabetes mellitus and smoking between the two groups(all P>0.05). There were statistically significant differences between the epilepsy group and non epilepsy group in hypertension(15 cases vs.22 cases), location of responsible aneurysms in middle cerebral artery(22 cases vs.24 cases), number of aneurysms(23 cases vs.41 cases), intracranial hematoma(15 cases vs.26 cases), hydrocephalus(15 cases vs.21 cases) and neurological sequelae(14 cases vs.20 cases)(χ 2=9.491, 23.840, 11.113, 6.737, 10.306, 9.161, all P<0.05). The results of multivariate analysis showed that hypertension, middle cerebral artery, multiple aneurysms, intracranial hematoma, hydrocephalus and neurologic sequelae were risk factors for epilepsy secondary to aneurysmal subarachnoid hemorrhage in the elderly[ OR(95% CI)2.361(1.476-3.421), 3.012(1.935-1.845), 1.494(1.027-1.845), 2.785(1.684-3.982), 1.920(1.283-2.984), 1.637(1.171-2.316)]. Conclusion:There are many factors influencing secondary epilepsy in elderly patients with aneurysmal subarachnoid hemorrhage.In order to reduce the incidence of secondary epilepsy, preventive measures should be taken against the above risk factors.

OBJECTIVETo investigate the quality of life of rectal cancer patients after anterior resection and the role of 5-hydroxytryptamine(5-HT) in the pathogenesis of anterior resection syndrome (ARS).

METHODSBetween November 2012 and October 2014, 90 rectal cancer patients who underwent Dixon procedure in the Institute of Surgery Research, Daping Hospital, Third Military Medical University, and developed ARS postoperatively were enrolled in the study. By clinic interview and telephone follow-up, they were investigated according to the 4 questionnaires, including gastrointestinal quality of life index(GIQLI), Wexner constipation and incontinence score(WCS, WIS), and 36-item short form health survey(SF-36). Associated clinical data and above parameters were compared between postoperative 0-6 months and 7-24 months. Expression of 5-HT in rectal mucosa was determined by immunohistochemistry preoperatively and postoperatively.

RESULTSThe GIQLI, WCS and WIS were significantly improved in 7-24 months group compared with those in 0-6 months group(all P<0.05). Furthermore, the SF-36 test result also showed significant improvement in the terms of physical function, physical role, vitality, social function, emotional role and health changes spheres in 7-24 months group(P<0.05). 5-HT expression in rectal mucosa(upper anastomosis 3402.95±1876.24, lower anastomosis 3045.35±1373.59 of ARS patients was significantly down-regulated compared with the preoperative expression(rectal margin mucosa 7176.60±3927.61)(P<0.05).

CONCLUSIONSPatients with ARS experience a significant trend toward recovery in their whole long-term quality of life. The down-regulation of 5-HT expression in rectal mucosa after surgery may be related with the pathogenesis of ARS.

Constipation ; Humans ; Intestinal Mucosa ; Prospective Studies ; Quality of Life ; Rectal Neoplasms ; Serotonin ; Treatment Outcome

Colorectal cancer (CRC) is the second most common cause of cancer-related death in the world. The pro-viral integration site for Moloney murine leukemia virus 1 (PIM1) is a proto-oncogene and belongs to the serine/threonine kinase family, which are involved in cell proliferation, migration, and apoptosis. Fibroblast growth factor receptor 1 (FGFR1) is a tyrosine kinase that has been implicated in cell proliferation, differentiation and migration. Small molecule HCI-48 is a derivative of chalcone, a class of compounds known to possess anti-tumor, anti-inflammatory and antibacterial effects. However, the underlying mechanism of chalcones against colorectal cancer remains unclear. This study reports that HCI-48 mainly targets PIM1 and FGFR1 kinases, thereby eliciting antitumor effects on colorectal cancer growth in vitro and in vivo. HCI-48 inhibited the activity of both PIM1 and FGFR1 kinases in an ATP-dependent manner, as revealed by computational docking models. Cell-based assays showed that HCI-48 inhibited cell proliferation in CRC cells (HCT-15, DLD1, HCT-116 and SW620), and induced cell cycle arrest in the G2/M phase through modulation of cyclin A2. HCI-48 also induced cellular apoptosis, as evidenced by an increase in the expression of apoptosis biomarkers such as cleaved PARP, cleaved caspase 3 and cleaved caspase 7. Moreover, HCI-48 attenuated the activation of downstream components of the PIM1 and FGFR1 signaling pathways. Using patient-derived xenograft (PDX) murine tumor models, we found that treatment with HCI-48 diminished the PDX tumor growth of implanted CRC tissue expressing high protein levels of PIM1 and FGFR1. This study suggests that the inhibitory effect of HCI-48 on colorectal tumor growth is mainly mediated through the dual-targeting of PIM1 and FGFR1 kinases. This work provides a theoretical basis for the future application of HCI-48 in the treatment of clinical CRC.