Objective To investigate the effect of exosomal miR-146a-5p expression on gray matter volume in patients with major depressive disorder(MDD).Methods A total of 113 MDD patients(MDD group)and 107 healthy controls(HC)(HC group)were selected.Peripheral blood was collected and exosomes were isolated to quantify miR-146a-5p expression.Brain high-resolution T1 WI images of MDD and HC were obtained via MR,and gray matter volume was computed via SPM12 software.The interaction effect of"Depression×miR-146a-5p expression"on gray matter volume was analyzed using SPM's Flexible factorial design,and the between-group difference was assessed by extracting the mean value,thus to analyze whether MDD-related gray matter volume abnormalities were dependent on miR-146a-5p expression.Results Exosomal miR-146a-5p expression was significantly elevated in MDD group compared to HC group.Voxel-based factorial analysis revealed a relationship between high miR-146a-5p expression in MDD group and reduced gray matter volume in the anterior and posterior cingulate cortices(independent voxel threshold P＜0.001,AlphaSim corrected),and a significantly reduced gray matter volume as compared with HC group was detected in the two regions.Conclusion The exosomal miR-146a-5p is overexpressed in patients with MDD and may be associated with specific cortical atrophy in patients with MDD.

Objective:To investigate the learning curve of laparoscopic transanal total mesorectal excision (taTME) for rectal cancer operated by one or two surgery teams.Methods:The retrospective cross-sectional study was conducted. Based on the concept of real-world research, the clinical data of 1 458 patients undergoing laparoscopic rectal cancer taTME from 44 medical centers who were registered in the Chinese taTME registry collaborative (CTRC) database from May 2010 to May 2020 were collected. The 1 458 patients were divided into cohorts with one surgery team or two surgery teams according to the operation method. Patients with one surgery team underwent taTME by transabdominal operation and then by transanal operation. Patients with two surgery teams underwent taTME by transabdominal and transanal operation simultaneously with duration of the simutaneous operation time ≥30 minutes. The entire surgical process of patients with two surgery teams is not required to be performed by two surgery teams simutaneously. The clinical data were collected from the medical centers with similar operation amount according to the operation time sequence to analyze the difference between different operation stages and explore the learning curve. The operation time was taken as the parameter to carry out cumulative sum analysis and draw the learning curve of laparoscopic rectal cancer taTME in each medical center. The clinicopathological characteristics of patients from two medical centers with the largest difference in learning curves were analyzed. Observation indicators: (1) screening results of clinical data; (2) clinical data collection of patients with one surgery team; (3) surgical situations of laparoscopic rectal cancer taTME from the one surgery team in different operation stages; (4) learning curve of the one surgery team; (5) clinical data collection of patients with two surgery teams; (6) surgical situations of laparoscopic rectal cancer taTME from the two surgery teams; (7) learning curve of the two surgery teams. The cumulative sum was calculated by the CUSUM=∑i=1nXi-U, where Xi represented the operation time of each taTME, U represented the average operation time of all cases, and n represented the operation number. Fitting process was conducted on scatter plot of learning curves. Taking the apex of learning curve as the boundary, the learning curve was divided into two stages. The abscissa corresponding to the apex of learning curve was the number of operations that needed to be performed to cross the learning curve. Measurement data with normal distribution were represented as Mean±SD. Comparison between two groups was conducted using the t test and comparison between multiple groups was conducted using the ANOVA. Measurement data with skewed distribution were represented as M( P25,P75), and comparison between groups was conducted using the Mann-Whitney U test. Comparison of ordinal data was analyzed using the rank sum test. Count data were analyzed using the chi-square test or Fisher exact probability. Results:(1) Screening results of clinical data:the clinical data of 661 patients from 7 medical centers with one surgery team and two surgery teams were collected. (2) Clinical data collection of patients with one surgery team: the clinical data of 312 patients undergoing laparoscopic rectal cancer taTME from 5 medical centers were collected including 42 cases in the number 2 medical center, 97 cases in the number 20 medical center, 82 cases in the number 33 medical center, 35 cases in the number 37 medical center and 56 cases in the number 39 medical center, respectively. (3) Surgical situations of laparoscopic rectal cancer taTME from the one surgery team in different operation stages: three medical centers including the number 2, number 37 and number 39 medical center with close operation volume provided the clinical data of cases distributed in five operation stages. Among the five operation stages, the proportion of high-quality operation of total mesorectal excision (TME) was ≥17/18, the incidence of postoperative complications was ≤13.3%(4/30) and the incidence of anastomotic leakage was ≤10.0%(3/30). There was no significant difference in the TME quality, postoperative complications or anastomotic leakage among the five operation stages ( P>0.05). There was no significant difference in the operation time among the five operation stages ( χ2=6.950, P>0.05). (4) Learning curve of the one surgery team: the number of operations corresponding to the turning point of learning curve in number 2 and number 20 medical center was 22 and 39, respectively. The number of operations corresponding to the turning points of learning curve in number 33 and number 37 medical center was 15, 66 and 10, 28, respectively. The number of operations corresponding to the turning point of learning curve in number 39 medical center was 20. The overall curve of number 20 medical center was in line with the trend of learning curve and 39 cases of operations was the minimum number needed to cross the learning curve. The biggest difference in learning curve was shown between the number 20 and number 33 medical center. Cases with the gender of male or female, age, body mass index, cases classified as stage 1, stage 2, stage 3 or stage 4 of the American Society of Anesthesiologists (ASA) Classification, cases with neoadjuvant therapy, duration of postoperative hospital stay of the number 20 medical center were 77, 20, (60±10)years, 24 kg/m 2(22 kg/m 2, 26 kg/m 2), 1, 88, 8, 0, 8, 8, 11 days (9 days, 13 days), respectively, versus 51, 31, (64±11)years, 23 kg/m 2(21 kg/m 2, 26 kg/m 2), 0, 35, 43, 1, 31, 16 days (13 day, 21 day) of number 33 medical center, showing significant differences in the above indicators between the two medical centers ( χ2 =6.442, t=-2.265, Z=-2.032, -6.870, χ2 =22.120, Z=-8.408, P<0.05). (5) Clinical data collection of the two surgery teams: the clinical data of 259 patients undergoing laparoscopic rectal cancer taTME from 5 medical centers were collected, including 46 cases in the number 2 medical center, 47 cases in the number 8 medical center, 78 cases in the number 18 medical center, 43 cases in the number 33 medical center and 45 cases in the number 44 medical center, respectively. (6) Surgical situations of laparoscopic rectal cancer taTME from the two surgery teams: four medical centers including the number 2, number 8, number 33 and number 44 medical center with close operation volume provided the clinical data of cases distributed in four operation stages. Among the four operation stages, the proportion of high-quality operation of TME was ≥50.0%(13/26), the incidence of postoperative complications was ≤35.0%(14/40) and the incidence of anastomotic leakage was ≤22.5%(9/40). There was no significant difference in the TME quality, postoperative complications or operation time among the four operation stages ( χ2 =3.252, 4.733, 8.848, P>0.05). There was a significant difference in the incidence of anastomotic leakage among the four operation stages ( P<0.05). (7) Learning curve of the two surgery teams: the number of operations corresponding to the turning point of learning curve in number 2 and number 8 medical center was 28 and 16, respectively. The number of operations corresponding to the turning points of learning curve in number 18, number 33 and number 44 medical center was 12 and 58, 10 and 36, 14 and 36, respectively. The overall curve of number 2 medical center was in line with the trend of learning curve and 28 cases of operations was the minimum number needed to cross the learning curve. The biggest difference in learning curve was shown between the number 2 and number 33 medical center. The age and cases with tumor in stage T0 and (or) Tis, stage T1, stage T2, stage T3 or stage T4 of the T staging of the number 2 and number 33 medical center were (60±12)years, 3, 1, 9, 11, 20 and (65±10)years, 2, 3, 22, 15, 0, respectively, showing significant differences in the above indicators between the two medical centers ( t=-2.280, Z=-4.033, P<0.05). Conclusion:Thirty-nine cases of operations was the minimum number for the one surgery team to cross the learning curve of laparoscopic rectal cancer taTME and 28 cases of operations was the minimum number for the two surgery teams to cross the learning curve of laparoscopic rectal cancer taTME.

Objective:To investigate the short term efficacy of laparoscopic assisted transanal total mesorectal excision (taTME) for low rectal cancer.Methods:The prospective study was conducted. The clinicopathological data of 80 patients who underwent laparoscopic assisted taTME for low rectal cancer in 8 medical centers，including 27 cases in the First Affiliated Hospital of Jilin University，16 cases in the Daping Hospital of Army Medical University，15 cases in the Beijing Friendship Hospital of Capital Medical University，10 cases in the Peking University Cancer Hospital，7 cases in the Peking Union Medical College Hospital of Chinese Academy of Medical Sciences，2 cases in the Peking University People′s Hospital，2 cases in the Liaoning Cancer Hospital Institute，1 case in the Ruijin Hospital of Shanghai Jiaotong University School of Medicine，from August 2017 to September 2018 were collected. Observation indicators：(1) clinical data of enrolled patients；(2) surgical situations；(3) postoperative histopathological examination；(4)postoperative complications and hospitalization. Measurement data with skewed distribution were represented as M(range). Count data were described as absolute numbers and (or) percentages. Results:(1) Clinical data of enrolled patients：a total of 80 patients were selected for eligibility. There were 59 males and 21 females，aged from 53 to 79 years，with a median age of 61 years. (2）Surgical situations：all 80 patients underwent surgery successfully，including 73 cases undergoing low anterior resection，4 cases undergoing Hartmann operation，1 case undergoing intersphincteric and abdominoperineal resection，1 case undergoing other operations and 1 case missing operation information. Nineteen of the 80 patients underwent transabdominal and transanal operations simultaneously. The operation time of 80 patients was 255 minutes (range，211?305 minutes). Of 80 patients，77 cases had the volume of intraoperative blood loss ≤500 mL，3 cases had the volume of intraoperative blood loss >500 mL，44 cases underwent instrumental anastomosis，24 cases underwent manual anastomosis，12 cases were missing anastomosis information，66 cases had specimens been taken out through anus，2 cases had specimens been taken out through Pfannens-tiel incision，10 cases had specimens been taken out through other ways，2 cases were missing the information of specimens removal ways，57 cases underwent preventive stoma，32 cases under-went anal canal indwelling，30 cases underwent free of splenic flexure and 2 cases were converted to open surgery. (3) Postoperative histopathological examination：of 80 patients，68 cases had the integrity of mesorectal specimens with complete，5 cases had the integrity of mesorectal specimens with near complete，1 case had the integrity of mesorectal specimens with not complete，6 cases were missing the information of integrity of mesorectal specimens，1 case had rectal perforation，1 case had positive circumferential margin and 1 case had positive distal margin. The number of lymph node dissected and diameter of tumor were 12(range，9?16) and 3.0 cm(range，1.9?4.0 cm) of 80 patients. Four of 80 patients achieved pathological complete remission. Cases with tumor stage as T0 stage，Tis stage，T1 stage，T2 stage，T3 stage or T4 stage of the pT staging，cases with tumor stage as N0 stage，N1 stage or N2 stage of the pN staging，cases with tumor stage as M0 stage or M1 stage of the pM staging were 4，2，11，24，35，4，55，21，4，75，5 of 80 patients. (4) Postopera-tive complications and hospitalization：8 of 80 patients underwent anastomotic leakage，including 2 cases with grade A anastomotic leakage，4 cases with grade B anastomotic leakage and 2 cases with grade C anastomotic leakage.Seven of 80 patients underwent intestinal obstruction. The 2 cases with grade A anastomotic leakage were improved after symptomatic drug treatment，the 4 cases with grade B anastomotic leakage were improved after treatment with antibiotics or catheter drainage and the 2 cases with grade C anastomotic leakage were improved after operation. The duration of hospital stay of 80 patients was 14 days(range，11?21 days). No patient died during hospitalization.Conclusion:Laparoscopic assisted taTME for low rectal cancer is safe and feasible，which has a good short term efficacy.

Objective:To study the relationship between zoledronic acid (ZOL) and vascular endothelial growth factor (VEGF) conformation so as to reveal the mechanism of bisphosphonates inhibiting angiogenesis.Methods:The binding structures of ZOL and VEGF were preprocessed and the molecular dockings were simulated through AutoDockTools, Discovery studio4 and AutoDockVina. The best binding conformation was accurately screened. The effects of various concentrations of ZOL (group A was 0 μmol/L, groups B, C and D were 25, 50 and 100 μmol/L, respectively) on human umbilical vein endothelial cell (HUVEC) proliferation, angiogenesis and angiogenic molecules were detected by using cell counting kit-8 (CCK-8) in vivo and in vitro angiogenesis, immunofluorescence and Western blotting. Results:There was a ZOL binding site on the target protein VEGF conformation. The affinity was -5.2 kcal/mol. This binding site consisted of the hydrophobic region composed of amino acids Cys26, 51, 57, etc. and the hydrogen bond binding region of the A chain (ASP34, SER50) and B chain (CYS61, 68, LEU66, GLY59). The results of CCK-8 showed that the levels of value A in groups B, C and D were significantly lower than that in group A at each time point from 3 to 6 days ( P<0.05). In vitro vascular experiments demonstrated that the numbers of budding in groups B, C and D [(208±28), (151±21) and (62±9), respectively] were significantly lower than that in group A (276±30) ( P<0.05). In vivo vascular experiments displayed that the ratio of Matrigel gel/plasma fluorescence in group A (0.003 1±0.000 3) was significantly higher than those in group B (0.002 1±0.000 2), group C (0.001 6±0.000 2) and group D (0.000 6±0.000 1) ( P<0.05). The results of Western blotting revealed that the expression of VEGF in groups B, C and D [(0.72±0.11), (0.41±0.07) and (0.24±0.04), respectively] were significantly lower than that in group A (1.01±0.02) ( P<0.05), and the expression levels of hypoxia-inducible factor-1α (HIF-1α) in groups B, C and D [(0.68±0.09), (0.55±0.06) and (0.43±0.08), respectively] were significantly lower than that in group A (0.96±0.04) ( P<0.05). Conclusions:ZOL could inhibit cell proliferation, in vivo and in vitro vascularization and expression of VEGF/HIF-1α. The binding site of ZOL with the conformation of VEGF was located in the hydrophobic region and hydrogen-bonding region of amino acids. Designing an antagonist targeting this site might potentially alleviate the effect of ZOL in inhibiting angiogenesis.

Objective:To nvestigate the clinical curative effects and advantages on neoadjuvant chemoradiation therapy (nCRT) combined with laparoscopy in lower rectal reserving annus surgery.Methods:we make a retrospective analysis of 141 patients (nCRT group) who received nCRT combined with laparoscopic rectal cancer resection and of 141 cases simply received laparoscopic rectal cancer resection at the same period in the department of coiorectal surgery,Cancer Hospital of China Medical University、Liaoning Cancer Hospital&Institute,from December 2013 to December 2015.Results:Two groups in operation time,anus exhausting time,hospital stay time,number of harvest lymph node,incision infection rate,anastomotic leakage rate are not statistic differences;Two groups of reserving anus rate (136/141 vs 125/141,P＜0.05),intraoperating bleeding (54.8 ± 23.4ml vs 69.7 ± 29.1 mL,P＜0.05),positive rate of distance resection margin (0/141VS 2/141,P＜0.05),length of distance resection margin(2.35 ± 0.70VS2.02 ± 0.46,P＜0.05),local recurrence rate (2/141 VS 9/141,P＜0.05) are statistic differences.Conclusion:Neoadjuvant chemoradiation therapy(nCRT) combined with laparoscopic rectal resection can improve reserving anus rate and reduce local recurrence rate.

Objective To compare the clinical effects of three-dimensional (3D) and two-dimensional (2D) laparoscopic surgeries for rectal cancer (RC) after neoadjuvant chemoradiation therapy (nCRT).Methods The retrospective cohort study was conducted.The clinicopathological data of 126 patients with RC who received laparoscopic surgery after nCRT in the Liaoning Cancer Hospital from January 2013 to January 2014 were collected.Of 126 patients,63 undergoing 3D laparoscopic surgery and 63 undergoing 2D laparoscopic surgery were respectively allocated into the 3D and 2D groups.Surgery was performed by the same doctors' team.Patients received surgery at 6-8 weeks after nCRT,and surgery followed the principle of total mesorectal excision.Observation indicators:(1) intra-and post-operative situations;(2) postoperative pathological examination;(3)follow-up and survival situations.Follow-up using outpatient examination and telephone interview was performed to detect local tumor recurrence and patients' survival up to January 2017.Patients received reexaminations once every 3 months within 1 year postoperatively and once every 6 months after 2-3 years postoperatively.Measurement data with normal distribution were represented as (x)±s and comparison between groups was analyzed using the t test.Comparisons of count data were analyzed using the chi-square test.Comparison of ordinal data was done by the nonparametric test.Survival curve was drawn using the Kaplan-Meier method.Survival was analyzed using the Logrank test.Results (1) Intra-and post-operative situations:all the patients underwent successful laparoscopic surgery for RC,without conversion to open surgery and perioperative death.Operation time,volume of intraoperative blood loss,time to anal exsufflation and number of patients with postoperative urinary dysfunction were (125±10)minutes,(54±23)mL,(44±5)hours,0 in the 3D group and (137±12)minutes,(62±20)mL,(46±5) hours,5 in the 2D group,respectively,with statistically significant differences between the 2 groups (t=5.777,2.038,2.575,x2 =7.138,P＜0.05).Number of patients with preserving anus,number of lymph node dissected,number of patients with anastomotic fistula and duration of hospital stay were 60,14.9±2.1,2,(9.5±0.8)days in the 3D group and 58,14.3± 1.7,4,(9.9±2.0)days in the 2D group,respectively,with no statistically significant difference between the 2 groups (x2 =0.133,t=1.606,x2 =0.175,t =1.329,P＞0.05).Two and 4 patients with anastomotic leakage in the 3D and 2D groups received defunctioning stoma,and finally anastomotic leakage healed,without anastomotic stenosis.Of 5 patients with urinary dysfunction in the 2D group,4 received indwelling catheter for 3 weeks and then can micturate autonomously after indwelling catheter removal,1 received indwelling catheter for 3 months and then can micturate autonomously after indwelling catheter removal,without suprapubic cystostomy.(2) Postoperative pathological examination:0 and 4 patients in the 3D and 2D groups had positive circumferential margin,with a statistically significant difference (x2=5.676,P＜0.05).One and 2 patients in the 3D and 2D groups had positive distal margin,with no statistically significant difference (x2 =1.606,P＞0.05).Number of patients with stage Ⅱ and Ⅲ of postoperative pathological staging were 30,33 in the 3D group and 32,31 in the 2D group,respectively,with no statistically significant difference between the 2 groups (x2=0.127,P＞0.05).(3) Follow-up and survival situations:126 patients were followed up for 36.0-48.0months,with a median time of 39.5 months.During the follow-up,0 and 3 patients in the 3D and 2D groups were complicated with local tumor recurrence,with a statistically significant difference between the 2 groups (x2 =4.232,P＜0.05).One-and 3-year overall survival rates were 98.4％,82.5％ in the 3D group and 96.8％,79.4％ in the 2D group,respectively,with no statistically significant difference between the 2 groups (x2 =0.206,P＞0.05).One-and 3-year disease-free survival rates were 92.7％,77.8％ in the 3D group and 90.5％,73.0％in the 2D group,respectively,with no statistically significant difference between the 2 groups (x2=0.421,P＞0.05).Conclusion Compared with 2D laparoscopic surgery,3D laparoscopic surgery for RC after nCRT is safe and feasible,it can also shorten operation time,reduce intraoperative bleeding,alleviate the influence of intestinal peristalsis function,protect pelvic nerves better and improve operation quality.

Objective To investigate the clinical effects and advantages of flexible 3D laparoscopic surgery on rectal cancer after neoadjuvant chemoradiotherapy (nCRT). Methods The data of 152 patients who received laparoscopic rectal cancer resection after nCRT excluding the cases of high rectal cancer, cardiac and pulmonary dysfunction were analyzed from January 2015 to January 2016 in the Department of Colorectal Surgery of Liaoning Cancer Hospital. The distances from the annal edge to the inferior tumor margin were within 8 cm in these patients. Among these patients, 76 cases received the 3D laparoscopic surgery after nCRT (3D-nCRT), and 76 cases undergone the 2D laparoscopic surgery after nCRT (2D-nCRT). Results Between two groups, the number of lymph node harvest (14.8±2.1 vs. 14.3±1.7), positive rate of the distal margin [1.3 % (1/76) vs. 2.6 % (2/76)], reserving anus rate [92.1 % (70/76) vs. 81.2 % (67/76)], local recurrence rate [1.3 % (1/76) vs. 3.9 % (3/76)] and anastomotic leakage rate [2.6 % (2/76) vs. 3.9 % (3/76)] had no statistical differences (all P>0.05), while the operative time [(125.3±10.2) min vs. (136.6±12.0) min], intraoperative bleeding [(54.1±23.2) ml vs. (61.9±19.5) ml], anus exhaust time [(43.5±5.0) h vs. (45.4±5.6) h] and positive rate of circumferential resection margin (CRM) [1.3 % (1/76) vs. 9.2 % (7/76)] had statistical differences (all P< 0.05). Conclusion Flexible 3D laparoscopic surgery on rectal cancer after nCRT can shorten operative time, reduce intraoperative bleeding, alleviate the influence of intestinal peristalsis function, and improve operative quality.

Objective:To establish the methods of isolated culture and functional identification of mice bone marrow derived tolerogenic dendritic cells (CD11b+F4/80 +TDCs) in vitro.Methods: Mice bone marrow cells were isolated and cultured to obtain iDCs with the simulation of mouse rmGM-CSF and rmIL-4.CD11b+F4/80 +TDCs were purified by fluorescence-activated cell sorting on day 6.The morphological changes of TDCs were observed with the inverted microscope dynamically .The expression of CD11b+F4/80 +TDCs were analyzed by the flow cytometry .Tolerogenic function of CD11b+F4/80 +TDCs was evaluated by the expression of MHCⅡ, CD83, IDO, TLR-2, IL-10 and TGF-β1.The expression of MHCⅡ was analyzed by the flow cytometry , and the expression of CD83, IDO and TLR-2 were analyzed by immune-histochemistry.The levels of IL-10 and TGF-β1 in the supernatant of CD11b+F4/80 +TDC were analyzed by ELISA .Meanwhile mature DCs ( mDCs) induced by LPS were used as control .Results:The fresh isolated bone marrow cells look like round and small under microscope .After two days of culture , cells became big and formed into clusters . Five or six days later, cells clusters increased, and the morphology of cells became irregular .At the same time, more dendrite ap-peared on the surface of cells .The percentage of CD11b+F4/80 +TDCs induced by rmGM-CSF and rmIL-4 was about 23%, and the purity of the purified BM CD11b+F4/80 +iDC was about 99%.Compared with mDCs, CD11b+F4/80 +TDCs expressed low levels of MHCⅡand CD83 and high levels of IDO, TLR-2, IL-10 and TGF-β1.Conclusion:CD11b+F4/80 +TDCs derived from mouse bone marrow could be induced successfully by rmGM-CSF and rmIL-4 in vitro.CD11b+F4/80 +TDCs showed tolerogenic function by the expressions of IL-10, TGF-β1, IDO and TLR-2.

Objective To analyze the incidence and clinical features of urothelial tumors in renal allograft recipients.Methods A retrospective analysis of 1042 patients received renal allografts who had taken immunosuppression for at least six months between 2006 and 2011 in The First Centre Hospital of Tianjin was performed.Results Eleven cases of uroepithelial tumors were diagnosed in the 1042 cases of renal transplantation ( 1.06％ ),of whom 9 cases were noticed by hematuria ( 81.8 ％ ),2 cases ( 18.2％ ) by medical examination.Six patients were diagnosed with multifocal urothelial carcinomas.Surgery was performed on all the patients with renal tumors and followed by chemotherapy or radiotherapy.Conclusion Malignancies in urinary tract after renal transplantation should be bore in mind.Early diagnosis is very important.The treatment options include reducing immunosuppressive agents and removing tumor lesions completely.