Objective:To investigate the effect of astragaloside IV (AS-IV) on the secretion of stromal cell-derived factor-1α (SDF-1α) and CXC chemokine receptor 4 (CXCR4) by high glucose injured human umbilical vein endothelial cells (HUVECs), so as to lay a foundation for further study on AS-IV improving angiogenesis by regulating SDF-1 α/CXCR4 axis of endothelial cells.Methods:HUVECs were isolated and cultured from the umbilical vein of full-term healthy newborns and identified by von Willebrand factor (vWF) combined with 4-diamino-2-phenylindole (DAPI) nuclear staining. The obtained HUVECs was cultured in EGM-2 medium with 30 mmol/L glucose for 120 h to obtain high glucose damaged HUVECs. After intervention with different concentration gradients (25 mg/L, 50 mg/L, 100 mg/L, 200 mg/L, 400 mg/L) AS-IV for 72 hours, the contents of SDF-1α and CXCR4 were detected by enzyme linked immunosorbent assay (ELISA) method to determine the best concentration of AS-IV. The supernatant of damaged HUVECs were collected at 6, 12, 24, 48 and 72 hours after intervention with the best concentration of AS-IV, and the contents of SDF-1α and CXCR4 were detected by ELISA method to determine the best action time of AS-IV. The damaged HUVECs was randomly divided into experimental group and control group, and the blank group was set up at the same time. The experimental group was treated with the best concentration of AS-IV and the best time, the control group and the blank group were treated with the same volume of phosphate buffered saline (PBS) solution, and the contents of SDF-1α and CXCR4 in each group were detected by ELISA method.Results:The vWF factor on the cell membrane was green fluorescence, and the nucleus was blue after DAPI staining. When the fusion image showed green fluorescence, HUVECs were identified by blue fluorescence. The expression of SDF-1α in damaged HUVECs was the best when treated with AS-IV of 100 mg/L for 24 hours (1 642.87 pg/ml), and the expression of CXCR4 in damaged HUVECs was the best when treated with AS-IV of 50 mg/L for 48 hours (8.44 ng/ml). Compared with the control group, the contents of SDF-1α and CXCR4 in the experimental group were significantly increased, and the difference was statistically significant ( P<0.05). While the contents of SDF-1α and CXCR4 in the experiment group were slightly less than those in the blank group and there was no statistically significant difference ( P>0.05). Conclusions:AS-IV can promote the expression of SDF-1α and CXCR4 in HUVECs damaged by high glucose to return to normal physiological level, so as to play the role of vascular repair and neovascularization.

Objective To investigate the association between KRAS gene mutations and clinicopathological characteristics and prognosis of colorectal cancer (CRC) patients.Methods The retrospective casecontrol study was conducted.The clinicophathological data of 315 patients who underwent radical resection of CRC in the Yangpu Hospital Affiliated to Tongji University between January 2007 and July 2011 were collected.Nextgeneration sequencing was performed to identify KRAS gene mutations from surgical specimens.Observation indicators:(1) detection of KRAS gene;(2) association between KRAS gene mutations and clinicopathological characteristics of CRC patients;(3) follow-up and survival situations;(4) multivariate analysis of KRAS gene mutations in the prognosis of CRC patients.Follow-up using outpatient examination and telephone interview was performed to detect postoperative overall survival up to August 2016.Comparisons of count data were analyzed using the chi-square test.Measurement data with skewed distribution were described as M (interquartile range),and comparison between groups was analyzed using the nonparametric test.The survival rate was calculated using the Kaplan-Meier method,and survival was compared using the Log-rank test.The multivariate analysis was done using the COX regression model.Results (1) Detection of KRAS gene:all the 315 patients finished gene detection of surgical specimens,including 172 in wide-type mutations and 143 in mutant-type mutations (mutations at codon 12 and 13 of KRAS exon 2 and other mutant points were respectively detected in 80,24 and 40 patients,and 1 patient had simultaneous mutations at codon 12 and 13 of KRAS exon 2;missense and nonsense mutations were respectively detected in 141 and 2 patients).The major point mutations were at p.G12D and p.G13D.(2) Association between KRAS gene mutations and clinicophathological characteristics of CRC patients:tumors located in the proximal colon,distal colon and rectum were respectively detected in 34,48,90 patients with wild-type mutation and in 44,27,72 patients with mutant-type mutation,with a statistically significant difference (x2 =0.038,P＜0.05).(3) Follow-up and survival situation:315 patients were followed up for 3-115 months,with a median time of 78 months.The postoperative overall survival rate was 41.0％ in 172 patients with wild-type KRAS mutations,27.4％ in 80 patients with KRAS codon 12 mutations,26.3％ in 24 patients with KRAS codon 13 mutations and 48.2％ in 40 patients with other KRAS mutations,showing a statistically significant difference (x2=0.040,P＜0.05).(4) Multivariate analysis of KRAS gene mutations in the prognosis of CRC patients:the results of multivariate analysis showed that mutations at codon 12 of KRAS exon 2 was an independent factor affecting poor prognosis of CRC patients (Hazard ratio=1.543,95％ confidence interval:1.050-2.265,P＜0.05).Conclusions Most KRAS mutations of CRC patients are at codon 12 and 13 of KRAS exon 2,and the major point mutations are at p.G12D and p.G13D.KRAS gene mutations may be associated with tumor location.Mutations at codon 12 of KRAS exon 2 is an independent factor affecting poor prognosis of CRC patients.

Objective:To investigate the effects of mesenchymal stem cells-derived exosomes (MSC-Exos) secreted by mesenchymal stem cells (MSCs) induced by astragaloside IV (AS-IV) on the biological function and pyroptosis of human umbilical vein endothelial cells (HUVECs) injured by high glucose.Methods:After human umbilical cord blood mesenchymal stem cells (hUCBMSCs) were intervened with 400 mg/L of AS-IV, exosomes were extracted, and then the morphology and specific markers of exosomes were identified. Human umbilical vein endothelial cells (HUVECs) were cultured in a medium with a glucose concentration of 30 mmol/L to prepare a high glucose-impaired HUVECs model. High glucose-impaired HUVECs were randomly divided into experimental and model groups, with the experimental group intervened with 100 μg/ml of MSC-Exos and the model group intervened with an equal volume of PBS solution, while a blank control group was also set up. Cell counting Kit-8 (CCK-8) cell proliferation assay, adhesion assay, matrigel tube formation assay and scratch assay were used to detect the effects of AS-IV-mediated MSC-Exos on the proliferation, adhesion, tube formation and migrationability of HUVECs; Western blot and real time fluorescence quantitative polymerase chain reaction (qRT-PCR) were used to detect the protein and mRNA expression of scorch death-related molecules, such as Caspase-1, GSDMD (Gasdermin D) and NLRP3 in each group.Results:The proliferation, adhesion number, tube number and migration width of HUVECs cells were significantly lower than those in the blank group ( P<0.05); The expression of Caspase-1, GSDMD, NLRP3 protein and their mRNA increased significantly ( P<0.001); Under the intervention of MSC-Exos mediated by AS-IV, the cell proliferation, adhesion number, tube number and migration width of HUVECs were significantly higher than those in the model group ( P<0.05); The expression of Caspase-1, GSDMD, NLRP3 protein and their mRNA decreased, with statistically significant difference ( P<0.05). Conclusions:AS-IV mediated MSC-Exos can significantly improve the biological function of high glucose-impaired endothelial cells, and its mechanism may be related to anti-pyroptosis.

OBJECTIVETo evaluate the preliminary efficacy of video-assisted anal fistula treatment (VAAFT) for complex anal fistula.

METHODSClinical data of 11 consecutive patients with complex anal fistula undergoing VAAFT in our department from May to July 2015 were reviewed. VAAFT was performed to manage the fistula under endoscope without cutting or resection.

RESULTSVAAFT was successfully performed in all the 11 patients. The internal ostium was closed using mattress suture in 10 cases, and Endo-GIA stapler in 1 case. The mean operative time was (42.0±12.4) min, mean hospital stay was (4.1±1.5) d. Complication included bleeding and perianal infection in 1 case respectively. After 1 to 3.2 months follow-up, success rate was 72.7%(8/11), and no fecal incontinence was observed.

CONCLUSIONVideo-assisted anal fistula treatment is an effective, safe and minimally invasive surgical procedure for complex anal fistula with preservation of anal sphincter function.

Fecal Incontinence ; Humans ; Length of Stay ; Minimally Invasive Surgical Procedures ; Operative Time ; Rectal Fistula ; Sutures

The unique advantages of traditional Chinese medicine (TCM) in treating late onset hypogonadism in male have gradually emerged with the continuous deepening of the understanding and research on late onset hypogonadism in male. Time sequence is a general summary of the natural growth and operational laws. Tian gui and testosterone have their normal time sequences, and they may be associated with each other. A man′s tian gui follows the regular time sequence from " inception" to " exhaustion" throughout " eight" under normal physiological conditions. " Tian gui out of time sequence" includes the loss of tian gui exuberance (dysfunction of viscera dominated by the liver) and exhaustion in the time sequence (pathological deficiency of viscera dominated by the kidney), resulting in " tian gui exhaustion" in advance of " eight eight". Tian gui and testosterone are key concepts in Chinese and Western medicine for understanding late onset hypogonadism in male. The theory of " tian gui out of time sequence" may be closely related to the core pathogenesis of this condition, particularly in cases of liver depression and kidney deficiency. This study suggests that restoring the normal time sequence of tian gui while treating the liver and kidney simultaneously through time-sharing treatment should be effective. The use of Xiongcan Yishen Formula has shown promising therapeutic result, offering new insights and references for treating late onset hypogonadism in male using TCM.

Objective: To perform an anatomical observation on the extension of the mesocolon to the mesorectum and the continuity of the fasciae lining the abdomen and pelvis, in order to clarify the appropriate surgical plane of total mesorectal excision. Methods: This is an descriptive study. The operation videos of 61 cases (28 males, 33 females, median age of 61) were collected. All the patients underwent laparoscopic colorectal surgery from January 2018 to December 2018 in Yangpu Hospital, including low anterior resection for rectal cancer in 25 cases, left hemicolectomy for descending colon cancer in 15 cases, and subtotal resection of the colon for intractable constipation in 21 cases. Among these 21 constipation patients, 8 received additional modified Duhamel surgeries. Gross anatomy was performed on 24 adult cadavers provided by Department of Anatomy, Shanghai Jiaotong University School of Medicine, including 23 formalin-fixed and 1 fresh cadaver (12 males, 12 females). Sixty-one patients and 24 cadavers had no previous abdominal or pelvic surgical history. The anatomy and extension of fasciae related to descending colon, sigmoid colon and rectum, especially the morphology of Toldt fascia, and the continuities of mesocolon and mesorectum were observed carefully. The distribution characteristics of the fasciae and anatomical landmarks during laparoscopic surgery were recorded and described. Results: The anatomical study on 24 cadavers showed that visceral fascia was the densest connective tissue in the pelvic, posterolateral to the rectum, and stretched as a hammock to lift all pelvic organs. Among 61 patients undergoing laparoscopic surgery, 36 (59.0%) needed to free the left colon during operation, and Toldt fascia in the descending colon segment presented as potential, avascular and extensible loose connective tissue plane between the mesocolon and posterior Gerota fascia; 33 (54.1%) needed to free the rectum during operation, and Toldt fascia extended downward to pelvis as loose connective tissue between the fascia propria of the rectum and visceral fascia; the fascia propria of the rectum exposed completely in 32 (32/33, 97.0%) cases, which ran downward and fused with visceral fascia at the level of the fourth sacral vertebra. The anatomy of 24 cadavers also showed that fascia propria of the rectum fused with visceral fascia at the level of Waldeyer fascia. The fusion line of these two fasciae was supposed to be the extension of Waldeyer fascia. There were two avascular planes behind the rectum: one between the fascia propria of the rectum and visceral fascia, and the other between the visceral fascia and parietal fascia. In 8 constipation cases undergoing laparoscopic subtotal colon resection plus modified Duhamel operation, both mesocolon and mesorectum needed to be mobilized. It was obvious that the mesocolon of descending colon extended and became the mesocolon of sigmoid colon, and ran further into the pelvic and became the mesorectum. The colon fascia of descending colon served as the natural boundary of mesocolon extended downward as the fascia of sigmoid colon and the fascia propria of the rectum, respectively. Toldt fascia locating between mesocolon of descending colon and Gerota fascia extended to pelvis as the 'presacral space’ between the fascia propria of the rectum and visceral fascia. Gerota fascia in descending colon segment extended as urogenital fascia in sigmoid colon segment and visceral fascia in the pelvis, respectively. In the cadaver anatomy study, the visceral fascia served as a corridor carrying the hypogastric nerve, and ureter was observed in 23 (23/24, 95.8%) cases. The visceral fascia passed from posterior to anterior lateral of rectum, fusing with Denonvilliers fascia in a fan shape. The pelvic plexus located exactly external to the junction of visceral fascia and Denonvilliers fascia. Pelvic splanchnic nerves went through the parietal fascia toward to the inferolateral of the pelvic plexus. Conclusion Fascia propria of the rectum and the visceral pelvic fascia are two independent layers of fascia, and the TME surgical plane is between the fascia propria of the rectum and visceral pelvic fascia instead of between the visceral and the parietal pelvic fascia.