OBJECTIVES: To explore the effects of acupuncture combined with rehabilitation training for promoting transdifferentiation of astrocytes into neurons in mice after cerebral ischemia. METHODS: Male C57/BL6J mice were subjected to intracerebral microinjection of an adeno-associated virus carrying the GFAP promoter for NeuroD1 and Ngn2 overexpression in the astrocytes, followed 3 or 12 days later by electrocoagulation of the distal middle cerebral artery. After modeling, the mice were randomly divided into model group without interventions and intervention group treated with electroacupuncture at the acupoints Baihui (GV20), left Hegu (LI4), Neiguan (PC6), Zusanli (ST36), and Yanglingquan (GB34) 24 h after surgery. The mice in the intervention group were housed individually in cages with running wheels, and their activity was recorded every 24 h. Neurological function scores of the mice were assessed on the 1st, 14th, and 21st days after modeling. Transdifferentiation of astrocytes in the target brain regions was observed using double immunofluorescence staining. RESULTS: Compared with those in the model group, the mice receiving eletroacupuncture and rehabilitation training showed significant improvement of neurological deficits at 14 and 21 days after modeling. The GFAP promoter of the AAV2/5 vector specifically labeled the local astrocytes, and compared with that that in the model group, the number of AAV-positive cells colabeled with the neuronal marker DCX significantly increased after 14 days of electroacupuncture and rehabilitation intervention, and the number of AAV-positive cells colabeled with the neuronal marker NeuN significantly increased after 21 days of intervention. CONCLUSIONS In mice with cerebral ischemia, electroacupuncture and rehabilitation training can promote transdifferentiation of astrocytes into neurons in the ischemic brain region, and the efficiency of transdifferentiation is positively correlated with the improvement of motor function.
Animals ; Electroacupuncture ; Astrocytes/cytology* ; Cell Transdifferentiation ; Male ; Mice, Inbred C57BL ; Brain Ischemia/physiopathology* ; Mice ; Neurons/cytology* ; Doublecortin Protein

Objective To modify the method for establishing mouse models of middle cerebral artery occlusion(MCAO)-induced focal cerebral ischemia using electrocoagulation.Methods Forty-six C57/BL6J male mice were divided into MCAO model group(n=34)and sham-operated group(n=12).In the model group,MCAO was induced by permanent coagulation of the right middle cerebral artery(MCA)using a coagulator,and cerebral blood flow perfusion was monitored before and at 20 min and 1 day after modeling.Neurological deficits of the mice at 1,7,and 14 days after modeling were evaluated using Longa score,mNSS score,beam walking test,cylinder test and corner test.TTC staining was used to measure the cerebral infarct size,and Western blotting was performed to detect the expressions of BDNF,GFAP and DCX proteins in the ischemic cortex.Results The mice in the model group showed significantly reduced cerebral blood flow in the MCA on the ischemic side and obvious neurological deficits with increased forelimb use asymmetry on days 1,7 and 14 after modeling(P<0.05).In the cerebral cortex on the ischemic side of the model mice,the expressions of GFAP and DCX increased significantly at 1,7,and 14 days(P<0.05)and the expression of BDNF increased at 1 day after modeling ischemia(P<0.05).Conclusion We successfully prepared mouse models of MCAO using a modified method by changing the electrocoagulation location from the distal location of the junction between the MCA and the inferior cerebral vein to a 2 mm segment medial to the junction between the MCA and the olfactory bundle.

Objective To modify the method for establishing mouse models of middle cerebral artery occlusion(MCAO)-induced focal cerebral ischemia using electrocoagulation.Methods Forty-six C57/BL6J male mice were divided into MCAO model group(n=34)and sham-operated group(n=12).In the model group,MCAO was induced by permanent coagulation of the right middle cerebral artery(MCA)using a coagulator,and cerebral blood flow perfusion was monitored before and at 20 min and 1 day after modeling.Neurological deficits of the mice at 1,7,and 14 days after modeling were evaluated using Longa score,mNSS score,beam walking test,cylinder test and corner test.TTC staining was used to measure the cerebral infarct size,and Western blotting was performed to detect the expressions of BDNF,GFAP and DCX proteins in the ischemic cortex.Results The mice in the model group showed significantly reduced cerebral blood flow in the MCA on the ischemic side and obvious neurological deficits with increased forelimb use asymmetry on days 1,7 and 14 after modeling(P<0.05).In the cerebral cortex on the ischemic side of the model mice,the expressions of GFAP and DCX increased significantly at 1,7,and 14 days(P<0.05)and the expression of BDNF increased at 1 day after modeling ischemia(P<0.05).Conclusion We successfully prepared mouse models of MCAO using a modified method by changing the electrocoagulation location from the distal location of the junction between the MCA and the inferior cerebral vein to a 2 mm segment medial to the junction between the MCA and the olfactory bundle.

Objective:To investigate the influencing factors for the number of lymph node harvested after Da Vinci robotic and laparoscopic radical gastrectomy for gastric cancer.Methods:The retrospective case-control study was conducted. The clinicopathological data of 1 396 patients who underwent Da Vinci robotic or laparoscopic radical gastrectomy for gastric cancer in the First Affiliated Hospital of Nanchang University from December 2014 to July 2019 were collected. There were 991 males and 405 females, aged (60±11) years. Surgery using Da Vinci robotic system or laparoscopic system was completed according to patients' wishes. Cases with early gastric cancer underwent D 1+ lymphadenectomy and cases with advanced gastric cancer underwent standard D 2 lymphadenectomy. Observation indicators: (1) intraoperative situations; (2) postoperative situations; (3) influencing factors for the number of lymph node harvested after radical gastrectomy for gastric cancer; (4) follow-up and survival. Follow-up using outpatient examination or telephone interview was performed to detect survival of patients up to October 2020. Measurement data with normal distribution were represented as Mean± SD. Univariate analysis was done using the chi-square test or Fisher exact probability. Multivariate analysis was performed using Logistic regression model. The survival rate was calculated by Kaplan-Meier method. Results:(1) Intraoperative situations: all the 1 396 patients underwent radical gastrectomy, including 415 cases undergoing Da Vinci robotic radical gastrectomy and 981 cases undergoing laparoscopic radical gastrectomy. Thirty-five of the 1 396 patients were converted to open surgery, including 5 cases undergoing Da Vinci robotic radical gastrectomy and 30 cases undergoing laparoscopic radical gastrectomy. Of the 1 396 patients, 983 cases underwent distal gastrectomy, 400 cases underwent total gastrectomy and 13 cases underwent proximal gastrectomy, among which 597 cases underwent Billroth Ⅰ anastomosis, 385 cases underwent Billroth Ⅱ anastomosis, 401 cases underwent Roux-en-Y anastomosis and 13 cases underwent residual stomach-esophagus anastomosis. The operation time, volume of intraoperative blood loss and cases with intraoperative blood transfusion were (221±51)minutes, (201±81)mL, 24 of 415 cases undergoing Da Vinci robotic radical gastrectomy, and (196±42)minutes, (232±76)mL, 75 of 981 cases undergoing laparoscopic radical gastrectomy, respectively. (2) Postoperative situations: the time to postoperative first flatus, time to postoperative initial liquid food intake and duration of postoperative hospital stay of the 1 396 patients were (3.0±1.0) days, (4.2±1.5) days and (9.0±3.8) days, respectively. Two hundred and ten of the 1 396 patients had postoperative complications including 170 cases with grade Ⅰ-Ⅱ complications and 40 cases with grade Ⅲ-Ⅴ complications. Eight of the 210 patients with postoperative complications died of serious complica-tions and the other 202 cases were cured after symptomatic treatment. Results of postoperative histopathological examination showed that there were 958 cases of adenocarcinoma, 220 cases of mucinous adenocarcinoma, and 218 cases of signet ring cell carcinoma. The number of lymph node harvested and the number of positive lymph node of the 1 396 patients were 26.0±8.3 and 3.6±0.9, respectively, and cases with the number of lymph node harvested ≥16 or <16 were 1 312 and 84. (3) Influencing factors for the number of lymph node harvested after radical gastrectomy for gastric cancer: results of univariate analysis showed that the operating surgeon, operation method, range of gastric resection, nerve invasion, degree of tumor invasion and tumor pathological N stage were related factors influencing the number of lymph node harvested after Da Vinci robotic and laparoscopic radical gastrectomy for gastric cancer ( χ2=13.167, 6.029, 15.686, 5.573, 9.402, 17.139, P<0.05). Results of multivariate analysis showed that the operating surgeon, operation method, range of gastric resection and tumor pathological N stage were independent factors influencing the number of lymph node harvested after Da Vinci robotic and laparoscopic radical gastrectomy for gastric cancer ( odds ratio=1.589, 2.018, 1.787, 0.267, 95% confidence interval as 1.221?2.068, 1.140?3.570, 1.066?2.994, 0.103?0.689, P<0.05). (4) Follow-up and survival: of the 1 396 patients, 1 256 cases were followed up for 2 to 70 months, with a median follow-up time of 27 months. The 3-year cumulative survival rate of the 1 256 cases was 70.2%. Conclusion:The operating surgeon, operation method, range of gastric resection and tumor pathological N stage are independent factors influencing the number of lymph node harvested after Da Vinci robotic and laparoscopic radical gastrectomy for gastric cancer.

Objective:To investigate the application value of enhanced recovery after surgery (ERAS) in totally Da Vinci robotic total gastrectomy.Methods:The retrospective cohort study was conducted. The clinicopathological data of 97 patients with gastric cancer who underwent totally Da Vinci robotic total gastrectomy in the First Affiliated Hospital of Nanchang University between January 2016 and February 2019 were collected.There were 57 males and 40 females, aged (59±10)years, with a range from 35 to 60 years. Of the 97 patients, 52 receiving perioperative management using ERAS were allocated into ERAS group, and 45 receiving traditional perioperative management were allocated into traditional group. Observation indicators: (1) intraoperative situations; (2) postoperative situations. Measurement data with normal distribution were expressed as Mean± SD, and comparison between groups was analyzed using the t test. Count data were described as absolute numbers, and the chi-square test was used for comparison between groups. Repeated measurement data were analyzed by ANOVA. Comparison of ordinal datas was analyzed using the Mann-Whitney U test. Results:(1) Intraoperative situations: patients in the ERAS group and traditional group underwent totally Da Vinci robotic total gastrectomy for gastric cancer successfully. Cases with Roux-en-Y anastomosis or uncut Roux-en-Y anastomosis (methods of digestive reconstruction), operation time, volume of intraoperative blood loss for the ERAS group were 25, 27, (205±28)minutes, (176±80)mL, respectively, versus 21, 24, (199±31)minutes, (182±81)mL for the traditional group, showing no significant difference in the above indicators between the two groups ( χ2=0.02, t=1.00, 0.37, P>0.05). (2) Postoperative situations: time to first out-of-bed activities, time to first anal flatus, time to initial liquid food intake, time to abdominal drainage tube removal, cases with postoperative complications, the number of lymph node dissected, cases in stage Ⅰ, Ⅱ, Ⅲ of postoperative tumor staging, duration of postoperative hospital stay, hospitalization expenses were (1.85±0.29)days, (2.90±0.47)days, (2.53±0.28)days, (5.72±0.95)days, 6, 28±8, 4, 25, 23, (6.43±0.52)days, (60 222±3 888)yuan in the ERAS group and (3.04±0.39)days, (3.82±0.36)days, (4.24±0.30)days, (6.75±0.48)days, 5, (27±6)days, 3, 20, 22, (8.47±0.69)days, (64 197±3 369)yuan in the traditional group, respectively. There were significant differences in the time to first out-of-bed activities, time to first anal flatus, time to initial liquid food intake, time to abdominal drainage tube removal, duration of postoperative hospital stay and hospitalization expenses between the two groups ( t=17.19, 10.69, 29.02, 6.58, 16.57, 5.34, P<0.05). There was no significant difference in the postoperative complications, the number of lymph node dissected, or postoperative tumor staging between the two groups ( χ2=0.01, t=0.68, Z=-0.46, P>0.05). From 2 hours after anesthesia awakening to 48 hours after surgery, the visual analog pain scores were changed from 3.06±0.29 to 2.13±0.32 in the ERAS group, and from 4.11±0.74 to 3.26±0.42 in the traditional group, respectively, showing a significant difference in the changing trend between the two groups ( F=264.45, P<0.05). There was no death or readmission in the postoperative 30 days. Conclusions:ERAS applied in the totally Da Vinci robotic total gastrectomy is safe and effective, which is associated with faster gastrointestinal function recovery, shorter hospital stay, better pain control, and quicker recovery afer surgery.

An excellent assistant for robotic radical gastrectomy can play an important role in the operation, especially in a initial team. In robotic gastric cancer surgery, an excellent assistant should actively participate in the operation process, choose the appropriate trocar position according to patient's body habitus. Moreover, he should master various surgical instruments skillfully and switch instruments fluently to assist the surgeon to expose key parts during operation, and provide effective help in the operative details, so that the whole operation process can run more smoothly and the operation efficiency and quality will be greatly improved. The growth of the assistants needs constant practice and summary of experience. Meanwhile, the encouragement of the chief surgeon also plays a positive role in promoting the development of the assistants.

Objective:To compare the short- and long-term outcomes of robot-assisted and laparoscopic radical resection for mid-low rectal cancer.Methods:A prospective randomized controlled trial was conducted. A total of 130 patients with mid-low rectal cancer (inclusion criteria: age > 18 or ≤80 years old; pathological diagnosis of rectal adenocarcinoma by colonoscopy; distance from tumor to the anal verge ≤12 cm; no distant metastasis; cT1-3N0-1 or ycT1-3 after neoadjuvant radiotherapy and chemotherapy; suitable for laparoscopic and robotic surgery) at the Department of Colorectal Surgery of the First Affiliated Hospital of Nanchang University from October 2016 to September 2018 were prospectively enrolled. According to computer-generated random number method, patients were randomly divided into the robot group ( n=66) and laparoscopy group ( n=64), and underwent robot-assisted surgery or laparoscopic surgery respectively. Clinicopathological data of all the patients were collected and analyzed. The demographic parameters, short- and long-term outcomes were compared between two groups. Results:One patient in robot group whose postoperative sample was diagnosed as rectal adenoma by pathology was excluded. There were no statistically significant differences in age, sex, BMI, ASA classification, distance from tumor to the anal verge, serum CEA level, CA199 level between two groups (all P>0.05). Operations were successfully performed in all the patients without conversion to open operation. Robotic surgery was found to be associated with less intraoperative blood loss than laparoscopic surgery [(73.4±49.7) ml vs. (119.1±65.7) ml, t=-4.461, P<0.001], while there were no statistically significant differences in surgical procedures, operation time, time to first flatus, time to first liquid intake, time to removal of catheter or postoperative hospital stay between two groups (all P>0.05). Besides, there was no significant difference in the morbidity of postoperative complication between two groups [10.8% (7/65) vs. 12.5 (8/64), χ 2=4.342, P=0.720]. The median number of harvested lymph node in the robot group and the laparoscopy group was 15.7±6.2 and 13.8±6.1 ( t=1.724, P=0.087). There were no significant differences between two groups in tumor sample length, distance between proximal and distal resection margin, integrity grade of TME specimen, number of positive lymph nodes, postoperative pathological stage and tumor differentiation (all P>0.05). The distal resection margin of samples in two groups was all negative. One case in the robot group was found to have positive circumferential resection margin. The median follow up was 24 (9 to 31) months. In the robot group and the laparoscopy group, the 2-year overall survival rate was 95.4% and 90.6% respectively; the 2-year disease-free survival rate was 90.8% and 85.9% respectively, whose differences were not significant (both P>0.05). Conclusion:Robot-assisted radical resection for mid-low rectal cancer can achieve similar short-term and long-term outcomes of laparoscopic resection, while robot-assisted surgery can decrease blood loss during operation, leading to more precise practice in minimally invasive surgery.

An excellent assistant for robotic radical gastrectomy can play an important role in the operation, especially in a initial team. In robotic gastric cancer surgery, an excellent assistant should actively participate in the operation process, choose the appropriate trocar position according to patient's body habitus. Moreover, he should master various surgical instruments skillfully and switch instruments fluently to assist the surgeon to expose key parts during operation, and provide effective help in the operative details, so that the whole operation process can run more smoothly and the operation efficiency and quality will be greatly improved. The growth of the assistants needs constant practice and summary of experience. Meanwhile, the encouragement of the chief surgeon also plays a positive role in promoting the development of the assistants.

Objective:To compare the short- and long-term outcomes of robot-assisted and laparoscopic radical resection for mid-low rectal cancer.Methods:A prospective randomized controlled trial was conducted. A total of 130 patients with mid-low rectal cancer (inclusion criteria: age > 18 or ≤80 years old; pathological diagnosis of rectal adenocarcinoma by colonoscopy; distance from tumor to the anal verge ≤12 cm; no distant metastasis; cT1-3N0-1 or ycT1-3 after neoadjuvant radiotherapy and chemotherapy; suitable for laparoscopic and robotic surgery) at the Department of Colorectal Surgery of the First Affiliated Hospital of Nanchang University from October 2016 to September 2018 were prospectively enrolled. According to computer-generated random number method, patients were randomly divided into the robot group ( n=66) and laparoscopy group ( n=64), and underwent robot-assisted surgery or laparoscopic surgery respectively. Clinicopathological data of all the patients were collected and analyzed. The demographic parameters, short- and long-term outcomes were compared between two groups. Results:One patient in robot group whose postoperative sample was diagnosed as rectal adenoma by pathology was excluded. There were no statistically significant differences in age, sex, BMI, ASA classification, distance from tumor to the anal verge, serum CEA level, CA199 level between two groups (all P>0.05). Operations were successfully performed in all the patients without conversion to open operation. Robotic surgery was found to be associated with less intraoperative blood loss than laparoscopic surgery [(73.4±49.7) ml vs. (119.1±65.7) ml, t=-4.461, P<0.001], while there were no statistically significant differences in surgical procedures, operation time, time to first flatus, time to first liquid intake, time to removal of catheter or postoperative hospital stay between two groups (all P>0.05). Besides, there was no significant difference in the morbidity of postoperative complication between two groups [10.8% (7/65) vs. 12.5 (8/64), χ 2=4.342, P=0.720]. The median number of harvested lymph node in the robot group and the laparoscopy group was 15.7±6.2 and 13.8±6.1 ( t=1.724, P=0.087). There were no significant differences between two groups in tumor sample length, distance between proximal and distal resection margin, integrity grade of TME specimen, number of positive lymph nodes, postoperative pathological stage and tumor differentiation (all P>0.05). The distal resection margin of samples in two groups was all negative. One case in the robot group was found to have positive circumferential resection margin. The median follow up was 24 (9 to 31) months. In the robot group and the laparoscopy group, the 2-year overall survival rate was 95.4% and 90.6% respectively; the 2-year disease-free survival rate was 90.8% and 85.9% respectively, whose differences were not significant (both P>0.05). Conclusion:Robot-assisted radical resection for mid-low rectal cancer can achieve similar short-term and long-term outcomes of laparoscopic resection, while robot-assisted surgery can decrease blood loss during operation, leading to more precise practice in minimally invasive surgery.

Objective To investigate the clinical efficacy of Da Vinci robotic and laparoscopic distal gastrectomy for gastric cancer.Methods The propensity score matching and retrospective cohort study was conducted.The clinicopathological data of 171 patients with gastric cancer who were admitted to the First Affiliated Hospital of Nanchang University from January 2015 to October 2016 were collected.There were 110 males and 61 females,aged from 38 to 81 years,with a median age of 57 years.Of 171 patients,70 undergoing Da Vinci robotic distal gastrectomy for gastric cancer and 101 undergoing laparoscopic distal gastrectomy were allocated into the robotic group and laparoscopic group,respectively.Observation indicators:(1) the propensity score matching conditions and comparison of general data between the two groups after the propensity score matching;(2) intraoperative and postoperative situations;(3) situations of pathological examination;(4) follow-up.Patients were followed up by outpatient examination and telephone interview to detect severe complications and survival after discharge up to October 2018.The overall survival time was from the operation data to end of follow-up or time of death.The propensity score matching was used to perform 1 ∶ 1 matching by Empower Stats.Measurement data with normal distribution were represented as Mean ± SD,and comparison between groups was done using the t test.Measurement data with skewed distribution were represented as M (range),and comparison between groups was done using the Mann-Whitney U test.Count data were represented as absolute number,and comparison between groups was analyzed using the chi-square test and comparison of ordinal data between groups was analyzed using the Mann-Whitney U test.The survival rate and curve were respectively calculated and drawn by the Kaplan-Meier method,and Log-rank test was used for survival analysis.Results (1) The propensity score matching conditions and comparison of general data between the two groups after the propensity score matching:124 of 171 patients had successful matching,including 62 in each group.The body mass index (BMI) and tumor diameter before matching were (24.2±2.4)kg/m2 and (50±13)mm in the robotic group,(25.1±2.1) kg/m2 and (45±14) mm in the laparoscopic group,showing statistically significant differences between the two groups (t =-2.676,2.045,P＜ 0.05).The BMI and tumor diameter after matching were (24.5 ± 2.3) kg/m2 and (49 ± 14) mm in the robotic group,(24.4 ± 2.2) kg/m2 and (48 ± 12) mm in the laparoscopic group,showing no statistically significant difference between the two groups (t=0.110,0.524,P＞0.05).(2) Intraoperative and postoperative situations:the total operation time,volume of intraoperative blood loss,level of C-reactive protein at day 1 postoperatively,level of C-reactive protein at day 3 postoperatively,volume of totally abdominal drainage were (147±13) minutes,(115±12)mL,(52.2±7.2)mg/L,(33.7±11.9)mg/L,353.5 mL (range,267.0-1 350.0 mL) in the robotic group,and (140± 12) minutes,(131 ± 12) mL,(58.2±7.4) mg/L,(41.1 ± 16.9) rag/L,397.0 mL (range,255.0-1 600.0 mL) in the laparoscopic group,respectively,showing statistically significant differences in the above indexes between the two groups (t =3.163,-7.814,-4.631,-2.840,Z =-4.351,P＜0.05).(3) Situations of pathological examination:patients after matching in the two groups received R0 resection,with negative duodenal margin and gastric margin.The number of lymph nodes dissected in the robotic group and laparoscopic group were 22±4 and 20±4,respectively,with a statistically significant difference between the two groups (t=2.812,P＜0.05).(4) Follow-up:124 patients after propensity score matching were followed up for 6-37 months,with a median time of 25 months.During the follow-up,no severe surgery-related complications such as obstruction of input or output loop and dumping syndrome were found in the two groups within 3 months after operation.The 2-year overall survival rate was 82.1％ and 75.2％ in the robotic and laparoscopic group,with no statistically significant difference between the two groups (x2 =0.436,P＞0.05).Conclusions Compared with laparoscopic surgery,Da Vinci robotic distal gastrectomy for gastric cancer has advantages in postoperative recovery and minimally invasion.There are similar 2-year overall survival rates in the two groups.