1.PE_PGRS37 protein promotes intracellular colonization of Mycobacterium tuberculosis by inhibiting macrophage autophagy flow
Mengyu LI ; Zhenjun ZHANG ; Tingting FENG ; Hui WANG ; Chanchan NIE ; Chunwen CHEN ; Yunjie GAO ; Yifan DUAN ; Ruonan GUO ; Yingying CUI ; Guanghui DANG ; Siguo LIU
Chinese Journal of Zoonoses 2025;41(10):1005-1010,1015
This study explored the effect of the Mycobacterium tuberculosis(Mtb)PE/PPE family protein PE_PGRS37 on the growth of Mycobacterium smegmatis(Ms)and macrophage autophagy during Mtb infection.The pe_pgrs37 gene was amplified from Mtb genome through PCR,and the recombinant vector pAIN-PE_PGRS37 was successfully constructed through homologous recombi-nation.pAIN-PE_PGRS37 and pAIN were integrated into Ms through electroshock to construct pAIN-PGRS37/Ms and pAIN/Ms re-combinant bacteria.Western blotting indicated that the PE_PGRS37 protein was correctly expressed in pAIN-PE_PGRS37/Ms.The re-combinant bacteria were inoculated in 7H9/7H10 medium,and their colony morphology and growth curves were observed.No signifi-cant difference in colony morphology was observed between pAIN-PE_PGRS37/Ms and pAIN/Ms.The growth rate significantly in-creased between 10 and 16 h,and a plateau was reached at 26 h.After infection of U937 cells with pAIN-PE_PGRS37/Ms and pAIN/Ms,macrophage autophagy flow was detected with western blotting and immunofluorescence.In the pAIN-PE_PGRS37/Ms-infected group,compared with the pAIN/Ms-infected group,macrophage LC3-II and p62 protein expression was significantly up-regulated(P<0.001)and inhibited autophagosome and lysosome fusion.The intracellular survival of the recombinant bacteria was detected through colony counting,and pAIN-PE_PGRS37/Ms showed significantly greater survival in macrophages at 12 h,24 h,and 48 h than pAIN/Ms(P<0.05).Our results suggested that PE_PGRS37 protein promotes Mycobacterium survival in macrophages by blocking macro-phage autophagy flow,thus inhibiting macrophage autophagy.
2.PE_PGRS37 protein promotes intracellular colonization of Mycobacterium tuberculosis by inhibiting macrophage autophagy flow
Mengyu LI ; Zhenjun ZHANG ; Tingting FENG ; Hui WANG ; Chanchan NIE ; Chunwen CHEN ; Yunjie GAO ; Yifan DUAN ; Ruonan GUO ; Yingying CUI ; Guanghui DANG ; Siguo LIU
Chinese Journal of Zoonoses 2025;41(10):1005-1010,1015
This study explored the effect of the Mycobacterium tuberculosis(Mtb)PE/PPE family protein PE_PGRS37 on the growth of Mycobacterium smegmatis(Ms)and macrophage autophagy during Mtb infection.The pe_pgrs37 gene was amplified from Mtb genome through PCR,and the recombinant vector pAIN-PE_PGRS37 was successfully constructed through homologous recombi-nation.pAIN-PE_PGRS37 and pAIN were integrated into Ms through electroshock to construct pAIN-PGRS37/Ms and pAIN/Ms re-combinant bacteria.Western blotting indicated that the PE_PGRS37 protein was correctly expressed in pAIN-PE_PGRS37/Ms.The re-combinant bacteria were inoculated in 7H9/7H10 medium,and their colony morphology and growth curves were observed.No signifi-cant difference in colony morphology was observed between pAIN-PE_PGRS37/Ms and pAIN/Ms.The growth rate significantly in-creased between 10 and 16 h,and a plateau was reached at 26 h.After infection of U937 cells with pAIN-PE_PGRS37/Ms and pAIN/Ms,macrophage autophagy flow was detected with western blotting and immunofluorescence.In the pAIN-PE_PGRS37/Ms-infected group,compared with the pAIN/Ms-infected group,macrophage LC3-II and p62 protein expression was significantly up-regulated(P<0.001)and inhibited autophagosome and lysosome fusion.The intracellular survival of the recombinant bacteria was detected through colony counting,and pAIN-PE_PGRS37/Ms showed significantly greater survival in macrophages at 12 h,24 h,and 48 h than pAIN/Ms(P<0.05).Our results suggested that PE_PGRS37 protein promotes Mycobacterium survival in macrophages by blocking macro-phage autophagy flow,thus inhibiting macrophage autophagy.
3.Application of clinical pathway teaching method in standardized training of general practitioners in gastroenterology
Meizhu YAN ; Min AN ; Manru SHEN ; Zhenjun GAO
Chinese Journal of Medical Education Research 2022;21(1):60-62
Objective:To investigate the efficacy of clinical pathway teaching method in standardized training of general practitioners in gastroenterology.Methods:From 2018 to 2019, 40 residents who participated in the standardized training of general practitioners in the Department of Gastroenterology, Zhongshan Hospital Qingpu Branch, Fudan University Medical College were randomized into the experimental group and the control group. The residents of the experimental group were trained by clinical pathway teaching method, while the control group were trained by traditional methods. After 6 weeks' teaching, theoretical examination, operation skills and case analysis test were assessed and satisfaction surveys were conducted.Results:The operation skills and case analysis scores of the experimental group were significantly better than those of the control group [operation test (91.50±2.77) vs. (89.80±3.74), P<0.01; case analysis (92.10±1.98) vs. (86.40±2.87), P<0.01]. The teaching satisfaction of the experimental group was significantly higher than that of the control group, and the difference was statistically significant ( P<0.01). Conclusion:The teaching model of clinical pathway can improve the teaching quality of the residents in the department of gastroenterology, improve the satisfaction of the doctors in the training, and broaden clinical thinking, which is worthy of promotion in clinical teaching.
4.Comparison between KeyPort access and traditional transanal endoscopic microsurgery in the treatment of rectal tumors
Zhigang GAO ; Xiaofeng HAN ; Yunlei WANG ; Jianwei ZHENG ; Yong YANG ; Zhenjun WANG
Chinese Journal of General Surgery 2020;35(5):353-356
Objective:To compare the efficacy and safety of KeyPort access and traditional transanal endoscopic microsurgery(TEM) in the treatment of rectal tumors.Methods:In this study, 36 cases of rectal tumors were treated by KeyPort TEM access and 52 cases by traditional TEM. Tumor type, size, distance from anal edge, operation time, intraoperative blood loss, postoperative hospitalization time, specimen quality and complications were compared between the two groups.Results:There were no significant differences in tumor type, size, operation time, intraoperative blood loss, postoperative hospitalization time, complications, recurrence and metastasis rate between the two groups. The distance between lower cutting edge to the anus in KeyPort access group was significantly greater than that of traditional TEM group[(6.7±1.9) vs. (5.1±1.8) cm , t=3.901, P<0.001]. All the surgeries in the KeyPort access group were completed. While two cases of in traditional TEM group were coverted to other surgical approaches. All patients in the KeyPort group had normal anal function in the early postoperative period, while 2 patients in the traditional TEM group suffered anal function impairment. Conclusion:TEM by KeyPort access is safer and more effective then traditional TEM, as well as more generous indications.
5. Application of KeyPort access to transanal endoscopic mircrosurgery
Zhigang GAO ; Xiaofeng HAN ; Yunlei WANG ; Yong YANG ; Zhenjun WANG
Chinese Journal of Gastrointestinal Surgery 2019;22(12):1131-1136
Objective:
To explore the effectiveness of KeyPort access in transanal endoscopic mircrosurgery (TEM).
Methods:
A descriptive case series study was performed. Clinicopathological data of 20 patients undergoing KeyPort access TEM in Beijing Chaoyang Hospital of Capital Medical University from December 2016 to April 2018 were collected and analyzed retrospectively. Procedure of KeyPort access TEM: general anesthesia or combined spinal epidural anesthesia (CSEA); lithotomy or prone jack-knife position; anal dilation; placement of the KeyPortaccess; connection of TEM pneumoperitoneum device, light source and imaging equipment; placement of 5 mm dedicated endoscope; insufflation of CO2 with pressure of 1.6-2.0 kPa (12-15 mmHg); after rinsing the intestinal lumen, circular resection marginlabeled by the needle-shaped electrocautery;electric coagulation or ultrasonic knife used to perform a full-thickness resection with a 0.5 cm-1 cm margin along the marking line. Indications of KeyPort access TEM: (1) benign large sessile polyps which were difficult to resect under colonoscopy; (2) submucosal lesions with diameter <2 cm; (3) Tis and T1 stage rectal carcinoma without lymph node metastasis; (4) palliative resection of T2 stage rectal carcinoma without lymph node metastasis. Contraindications: (1) accompanying serious diseases without the tolerance of anesthesia and operation; (2) distance from lesion to anal verge >20 cm.
Results:
There were 10 males and 10 females with age of (63±15) years old and BMI of (24.5±3.3) kg/m2. The diameter of the lesions was (2.0±1.3) cm, and the distance from lesion to anal verge was (6.2±2.2) cm. One patient had 3 lesions at different positions in rectum with diameters of 0.5 cm, 0.5 cm, and 1 cm, respectively. All operations were accomplished through the KeyPort access TEM and no case was transferred to other methods. The duration of surgery was 75 (30-220) minutes; intraoperative blood loss was 10 (0-30) ml. Two patients with rectal anterior wall lesions underwent full-thickness resection of the intestine wall reaching the peritoneal reflex with penetration into the peritonealcavity, and received suture closure immediately. For the patient with 3 rectal lesions, the 1.0 cm lesion received a full-thickness resection and the other 2 lesions received submucosal resection. No postoperative complication occurred. Postoperative pathology showed that there were 1 case of chronic inflammatory lesion, 4 cases of benign tumor, 3 cases of carcinoma in situ, 4 cases of neuroendocrine tumor, 6 cases of pT1 rectal cancer, 2 cases of pT2 rectal cancer (both invading the superficial muscle layer). The median hospital stay was 6 (3-7) days. The postoperative follow-up was (7.2±3.8) months. No postoperative complication or recurrence was observed.
Conclusion
TEM with KeyPort access is safe, rapid and effective in the treatment of rectal tumors.
6.Risk Analysis of Radiotherapy Implementation Process Based on Failure Mode and Effect Analysis.
Mingyin JIANG ; Linlin WANG ; Jiaqi GAO ; Mengya HU ; Qin LI ; Zhenjun PENG ; Qingmin FENG ; Xutian ZHANG ; Qiang ZHANG ; Shenglin LIU
Chinese Journal of Medical Instrumentation 2019;43(3):230-234
OBJECTIVE:
Providing a risk assessment method for the implementation of radiotherapy to identify possible risks in the implementation of the treatment process, and proposing measures to reduce or prevent these risks.
METHODS:
A multidisciplinary expert evaluation team was developed and the radiotherapy treatment process flow was drawn. Through the expert team, the failure mode analysis is carried out in each step of the flow chart. The results were summarized and the (risk priority ordinal) score was obtained, and the quantitative evaluation results of the whole process risk were obtained.
RESULTS:
One hundred and six failure modes were obtained, risk assessment of (20%) high risk failure model are 22 and severity (≥ 8) high risk failure model are 27. The reasons for the failures were man-made errors or hardware and software failures.
CONCLUSIONS
Failure mode and effect analysis can be used to evaluate the risk assessment of radiotherapy, and it provides a new solution for risk control in radiotherapy field.
Healthcare Failure Mode and Effect Analysis
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Risk Assessment
7.Application of KeyPort access to transanal endoscopic mircrosurgery
Zhigang GAO ; Xiaofeng HAN ; Yunlei WANG ; Yong YANG ; Zhenjun WANG
Chinese Journal of Gastrointestinal Surgery 2019;22(12):1131-1136
Objective To explore the effectiveness of KeyPort access in transanal endoscopic mircrosurgery (TEM). Methods A descriptive case series study was performed. Clinicopathological data of 20 patients undergoing KeyPort access TEM in Beijing Chaoyang Hospital of Capital Medical University from December 2016 to April 2018 were collected and analyzed retrospectively. Procedure of KeyPort access TEM: general anesthesia or combined spinal epidural anesthesia (CSEA); lithotomy or prone jack?knife position; anal dilation; placement of the KeyPortaccess; connection of TEM pneumoperitoneum device, light source and imaging equipment; placement of 5 mm dedicated endoscope; insufflation of CO2 with pressure of 1.6?2.0 kPa (12?15 mmHg); after rinsing the intestinal lumen, circular resection marginlabeled by the needle?shaped electrocautery;electric coagulation or ultrasonic knife used to perform a full?thickness resection with a 0.5 cm?1 cm margin along the marking line. Indications of KeyPort access TEM: (1) benign large sessile polyps which were difficult to resect under colonoscopy; (2) submucosal lesions with diameter <2 cm; (3) Tis and T1 stage rectal carcinoma without lymph node metastasis; (4) palliative resection of T2 stage rectal carcinoma without lymph node metastasis. Contraindications: (1) accompanying serious diseases without the tolerance of anesthesia and operation; (2) distance from lesion to anal verge>20 cm. Results There were 10 males and 10 females with age of (63±15) years old and BMI of (24.5±3.3) kg/m2. The diameter of the lesions was (2.0±1.3) cm, and the distance from lesion to anal verge was (6.2±2.2) cm. One patient had 3 lesions at different positions in rectum with diameters of 0.5 cm, 0.5 cm, and 1 cm, respectively. All operations were accomplished through the KeyPort access TEM and no case was transferred to other methods. The duration of surgery was 75 (30?220) minutes;intraoperative blood loss was 10 (0?30) ml. Two patients with rectal anterior wall lesions underwent full?thickness resection of the intestine wall reaching the peritoneal reflex with penetration into the peritonealcavity, and received suture closure immediately. For the patient with 3 rectal lesions, the 1.0 cm lesion received a full?thickness resection and the other 2 lesions received submucosal resection. No postoperative complication occurred. Postoperative pathology showed that there were 1 case of chronic inflammatory lesion, 4 cases of benign tumor, 3 cases of carcinoma in situ, 4 cases of neuroendocrine tumor, 6 cases of pT1 rectal cancer, 2 cases of pT2 rectal cancer (both invading the superficial muscle layer). The median hospital stay was 6 (3?7) days. The postoperative follow?up was (7.2±3.8) months. No postoperative complication or recurrence was observed. Conclusion TEM with KeyPort access is safe, rapid and effective in the treatment of rectal tumors.
8.Application of KeyPort access to transanal endoscopic mircrosurgery
Zhigang GAO ; Xiaofeng HAN ; Yunlei WANG ; Yong YANG ; Zhenjun WANG
Chinese Journal of Gastrointestinal Surgery 2019;22(12):1131-1136
Objective To explore the effectiveness of KeyPort access in transanal endoscopic mircrosurgery (TEM). Methods A descriptive case series study was performed. Clinicopathological data of 20 patients undergoing KeyPort access TEM in Beijing Chaoyang Hospital of Capital Medical University from December 2016 to April 2018 were collected and analyzed retrospectively. Procedure of KeyPort access TEM: general anesthesia or combined spinal epidural anesthesia (CSEA); lithotomy or prone jack?knife position; anal dilation; placement of the KeyPortaccess; connection of TEM pneumoperitoneum device, light source and imaging equipment; placement of 5 mm dedicated endoscope; insufflation of CO2 with pressure of 1.6?2.0 kPa (12?15 mmHg); after rinsing the intestinal lumen, circular resection marginlabeled by the needle?shaped electrocautery;electric coagulation or ultrasonic knife used to perform a full?thickness resection with a 0.5 cm?1 cm margin along the marking line. Indications of KeyPort access TEM: (1) benign large sessile polyps which were difficult to resect under colonoscopy; (2) submucosal lesions with diameter <2 cm; (3) Tis and T1 stage rectal carcinoma without lymph node metastasis; (4) palliative resection of T2 stage rectal carcinoma without lymph node metastasis. Contraindications: (1) accompanying serious diseases without the tolerance of anesthesia and operation; (2) distance from lesion to anal verge>20 cm. Results There were 10 males and 10 females with age of (63±15) years old and BMI of (24.5±3.3) kg/m2. The diameter of the lesions was (2.0±1.3) cm, and the distance from lesion to anal verge was (6.2±2.2) cm. One patient had 3 lesions at different positions in rectum with diameters of 0.5 cm, 0.5 cm, and 1 cm, respectively. All operations were accomplished through the KeyPort access TEM and no case was transferred to other methods. The duration of surgery was 75 (30?220) minutes;intraoperative blood loss was 10 (0?30) ml. Two patients with rectal anterior wall lesions underwent full?thickness resection of the intestine wall reaching the peritoneal reflex with penetration into the peritonealcavity, and received suture closure immediately. For the patient with 3 rectal lesions, the 1.0 cm lesion received a full?thickness resection and the other 2 lesions received submucosal resection. No postoperative complication occurred. Postoperative pathology showed that there were 1 case of chronic inflammatory lesion, 4 cases of benign tumor, 3 cases of carcinoma in situ, 4 cases of neuroendocrine tumor, 6 cases of pT1 rectal cancer, 2 cases of pT2 rectal cancer (both invading the superficial muscle layer). The median hospital stay was 6 (3?7) days. The postoperative follow?up was (7.2±3.8) months. No postoperative complication or recurrence was observed. Conclusion TEM with KeyPort access is safe, rapid and effective in the treatment of rectal tumors.
9.Application value of biological mesh in the pelvic floor reconstruction of extralevator abdominoperineal excision for advanced low rectal cancer
Jiagang HAN ; Zhenjun WANG ; Guanghui WEI ; Zhigang GAO ; Baocheng ZHAO ; Zhiwei ZHAI ; Bingqiang YI ; Yong YANG ; Huachong MA ; Zhulin LI ; Jianliang WANG ; Sanshui YU ; Liangang MA ; Weigen ZENG
Chinese Journal of Digestive Surgery 2018;17(2):161-167
Objective To investigate the application value of biological mesh in the pelvic floor reconstruction of extralevator abdominoperineal excision (ELAPE) for advanced low rectal cancer (RC).Methods The retrospective cohort study was conducted.The clinicopathological data of 228 patients with advanced low RC who underwent ELAPE in the Beijing Chaoyang Hospital of Capital Medical University between August 2008 and December 2016 were collected.Of 228 patients,174 using biological mesh closure and 54 using primary closure were respectively allocated into the biological mesh group and primary closure group.Observation indicators:(1)intra-and post-operative situations;(2) postoperative complications (including short-term and long-term complications);(3) follow-up and survival situations.Follow-up using outpatient examination and telephone interview was performed to detect postoperative long-term complications,tumor recurrence or metastasis and overall survival up to December,2017.Measurement data with normal distribution were represented as( x) ±s,and comparison between groups was analyzed using the independent-sample t test.Measurement data with skewed distribution were described as M (range),and comparison between groups was analyzed using the nonparametric test.Comparisons of count data were evaluated by the chi-square test or Fisher exact probability.Results (1)Intra-and post-operative situations:all the patients underwent successful ELAPE.The perineal operation time,time of indwelling perineal drainage-tube and hospital expenses were respectively (60 ± 50)minutes,(11.6 ± 2.4) days,(57 781± 11 337) yuan in the biological mesh group and (50±21) minutes,(8.9± 1.7) days,(53 714± 13 395)yuan in the primary closure group,with statistically significant differences between groups (t =3.327,7.691,-2.203,P<0.05).The total operation time and duration of postoperative hospital stay were respectively (242±53) minutes,(13.0±5.0) days in the biological mesh group and (228±51) minutes,(12.0±5.0) days in the primary closure group,with no statistically significant difference between groups (t =1.701,1.309,P>0.05).(2) Postoperative complications:26 and 19 patients in the biological mesh group and primary closure group had respectively perineal wound complications (1 patient combined with multiple complications),showing a statistically significant difference between groups (x2 =10.660,P<0.05).The perineal wound infection,perineal hernia and disruption of perineal wound were respectively detected in 20,6,1 patients in the biological mesh group and 12,7,3 patients in the primary closure group,showing statistically significant differences between groups (x2 =3.931,5.282,P<0.05).(3) Follow-up and survival situations:174 patients in the biological mesh group were followed up for 64 months (range,13-112 months),and 54 patients in the primary closure group were followed up for 51 months (range,23-76 months).The local recurrence rate,distal metastasis rate and overall survival rate were respectively 5.17% (9/174),20.11% (35/174),77.59% (135/174) in the biological mesh group and 7.41%(4/54),24.07%(13/54),79.63%(43/54) in the primary closure group,with no statistically significant difference between groups (x2 =0.080,0.389,0.101,P>0.05).Conclusions The biological mesh in the pelvic floor reconstruction of ELAPE for advanced low RC is safe and feasible.Compared with primary closure,biological mesh closure will extend perineal operation time and time of indwelling perineal drainage-tube,and increase hospital expenses,but doesn't affect total operation time and duration of postoperative hospital stay,meanwhile,it can also reduce the overall perineal wound complications,especially in perineal wound infection,perineal hernia and disruption of perineal wound.
10.Laparoscopy combined with transperineal extralevator abdominoperineal excision for locally advanced low rectal cancer.
Jiagang HAN ; Zhenjun WANG ; Zhigang GAO ; Guanghui WEI ; Yong YANG ; Bingqiang YI ; Zhiwei ZHAI ; Huachong MA ; Bo ZHAO ; Baocheng ZHAO ; Hao QU ; Jianliang WANG ; Zhulin LI
Chinese Journal of Gastrointestinal Surgery 2016;19(6):654-658
OBJECTIVETo evaluate the laparoscopy combined with transperineal extralevator abdominoperineal excision (TP-ELAPE) for locally advanced low rectal caner.
METHODSClinical data of 12 patients with locally advanced low rectal cancer undergoing laparoscopy combined with TP-ELAPE in our department from May 2013 to March 2015 were retrospectively analyzed. There were 8 male and 4 female patients with median aged of 63 (46 to 72) years. The median distance from tumor lower margin to anal verge was 3.5(2.0 to 4.0) cm. A self-made transanal suit for minimally invasive operation was used to make a sealed lacuna outside the sphincter, thus laparoscope can be applied to perform transperineal operation.
RESULTSAll the patients underwent operations successfully without conversion to open abdominal operation. The median operating time was 206 (180 to 280) minutes with perineal operating time 95(80 to 120) minutes. The median intraoperative blood loss was 120(50 to 200) ml. The median postoperative hospital stay was 12(9 to 18 ) days. Postoperative pathology revealed that all circumferential margins (CRM) were negative. The area of sample horizontal section was (2 824±463) mm(2), and of outer muscularis propria was(2 190±476) mm(2). Postoperative complications included chronic sacrococcygeal region pain in 2 cases, urinary retention in 3 cases, perineal wound infection in 1 case. No perineal seroma, perineal hernia, wound dehiscence and sinus tract formation were observed. Among 8 patients with preoperative normal sexual function, sexual dysfunction occurred in 2 patients. There was no local recurrence and metastasis during a median follow-up of 21(12 to 34) months.
CONCLUSIONLaparoscopy combined with TP-ELAPE has the potential to simplify the operation procedure for low rectal cancer, can ensure the radical treatment and safety of operation, and may be carried out in experienced centers.
Abdomen ; Aged ; Anal Canal ; Blood Loss, Surgical ; Digestive System Surgical Procedures ; methods ; Female ; Humans ; Laparoscopy ; Length of Stay ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; Operative Time ; Perineum ; Postoperative Complications ; Postoperative Period ; Rectal Neoplasms ; surgery ; Rectum ; Retrospective Studies

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