1.Preparation and identification of a 9B9 monoclonal antibody specifically targeting EGFRvⅢ/EGFR
Huiping GAO ; Hua JIANG ; Huamao WANG ; Zonghai LI
Chinese Journal of Cancer Biotherapy 1996;0(04):-
Objective:To prepare and identify monoclonal antibody specifically targeting epidermal growth factor receptor(EGFR) and(or) epidermal growth factor receptor vⅢ(EGFRvⅢ),and to investigate its inhibitory effects on human hepatocellular carcinoma Huh7-EGFRvⅢ cell-and epidermal carcinoma A431 cell-implanted tumors in nude mice.Methods: BALB/c mice were immunized with 3T3 cells stably transfected with EGFRvⅢ(3T3-EGFRvⅢ).Immunized spleen cells were fused with myeloma SP2/0 cells,and anti-EGFRvⅢ monoclonal antibody positive hybridoma cells(named 9B9 antibody and 9B9 cells,respectively) were selected and identified by ELISA.The specific interaction between 9B9 antibody and EGFRvⅢ/EGFR antigen was detected by Western blotting and immunofluorescence assay.Huh7-EGFRvⅢ cell-(human hepatocellular carcinoma Huh7 cells stably transfected with EGFRvⅢ) and epidermal cell carcinoma A431 cell-bearing mouse models were established and were divided into PBS group,Cetuximab group and 9B9 antibody group.Then,anti-tumor effect of 9B9 antibody was examined and compared with those of PBS and Cetuximab.Results: A monoclonal antibody,named 9B9 antibody,was obtained by hybridoma technique and it reacted with both EGFRvⅢ antigen and EGFR antigen as detected by Western blotting and immunofluorescence.The inhibitory rates of Cetuximab and 9B9 antibody against Huh7-EGFRvⅢ cells-implanted tumors were 42% and 46%,respectively,and those against A431 cells-implanted tumors were 85% and 86%,respectively.Conclusion: 9B9 monoclonal antibody can effectively inhibit the growth of human hepatocellular carcinoma cell-and epidermal cell carcinoma cell-implanted tumors,and the effects resemble that of Cetuximab.
2.Preliminary Study on the Effect of 5-Aza-CdR on the Demethylation of RNF180 in Prostate Cancer Cell Line DU145
Haiguang WANG ; Huamao JIANG ; Zhirun ZUO ; Huangzhe LONG ; Guanyuan YUAN ; Fanzhen JIA
Journal of China Medical University 2017;46(2):140-144
Objective To investigate the mechanism and cause of the inactivation of tumor suppressor gene RNF180 in prostate cancer cell line by observing the effect of 5-Aza-CdR on the RNF180 gene in prostate cancer cell line DU145. Methods MTT method was adopted to study the effect of 5-Aza-CdR(0,1,2,5,10,15 and 20μmoI/L)on the proliferation of prostate cancer cells. Western blotting,real-time PCR,and methyla-tion specific PCR(MSP)were separately used to detect the expression of RNF180 in prostate cancer cells before and after the treatment of the most suitable drug concentration(5μmoI/L). Results In a certain range,the effect of 5-Aza-CdR on the proliferation of prostate cancer cell line DU145 was increased with the increase of drug concentration and the time of drug treatment(P<0.05). After the treatment of the most suitable drug concentration,the protein and mRNA expression of RNF180 in prostate cancer cells was significantly increased(P<0.05),but the methyla-tion of the promoter region was obviously decreased. Conclusion 5-Aza-CdR can reverse the methylation status of RNF180 gene in DU145 pros-tate cancer cell line,and relieve the silencing status of RNF180gene expression.
3.Endoscopic treatment of ureteral fibroepithlial polyp
Chuanliang XU ; Xiaofei WEN ; Yinghao SUN ; Jianguo HOU ; Huamao YE ; Xu GAO ; Xiaofeng GAO ; Linhui WANG
Chinese Journal of Urology 2008;29(12):836-838
Objective To report the experience on the ureteroscopic treatment of ureteral fibro-epithlial polyp by Holmium:YAG laser resection.Methods Of five cases,the polyp was located in the upper 1 third of the ureter in 2 cases,and in middle 1 third or ureter in 2 cases,in lower 1 third of ureter in 1 case.The length of the polyps ranged from 3 to 16 cm.Three patients presented with flank pain,4 with hematuria and 1 with hydronephrosis.Five patients underwent rigid ureteroscopic treat-ment.TUR was performed in the 2 polyp cases with prolapsing from the ureteral orifice.A Holmium:YAG laser was used to resect ureteral polyps.At the end of the procedure,a 7 F double-J ureteral stent was placed and indwelling for 6- 8 weeks.Results All operations were successfully done.The pathologic diagnosis were fibroepithelial polyp.Histologically,the polyps were composed of a central fibrovascular core surrounded by hyperplastic benign urothelium.The stroma of polyp consis-ted of fibrous connective tissue with minimal cellular infiltration,and occasional epithelial cell nests were seen.The average length of hospital stay was 3 d (range 2 to 5).The mean follow-up was 24 months (range 3 to 51),and all patients remained no recurrence.One patient developed a ureteral stricture 3 months after the treatment,and relieved by endoscopic incision by Holmium:YAG laser.Conclusion Endoscopic management of ureteral fibroepithelial polyps could be a treatment modality with minimal morbidity and good treatment results.
4.RNF180 Promoter Methylation in Prostate Cancer
Haiguang WANG ; Huamao JIANG ; Zhirun ZUO ; Huangzhe LONG ; Guanyuan YUAN ; Fanzhen JIA
Journal of China Medical University 2017;46(6):561-565
Objective To clarify the significance of RNF180 expression in carcinogenesis and progression of prostate cancer by detection of RNF180 promoter methylation. Methods RNF180 expression was detected in human prostate cancer cell lines(PC3,LNCap,and DU145)and normal prostate cells(RWPE?1)via Western blotting,RT?PCR,methylation?specific PCR(MSP),bisulfite?sequeneing PCR(BSP),respectively, while RNF180 expression in human prostate cancer tissues and paired adjacent non?tumor tissue was detected via immunohistochemistry. Results The expressions of RNF180 mRNA and protein in prostate cancer cells were significantly lower than those in normal prostate cells(P<0.05),op?posite to what was observed for the methylation level of the RNF180 promoter. Additionally,the RNF180 expression in prostate cancer tissue was significantly lower than that in paired adjacent non?tumor tissue. Conclusion The RNF180 promoter is incompletely methylated in prostate can?cer cells,which may be a reason for the decline or silencing of RNF180 expression in cancer cells and tissues.
5.Research on primary and advanced levels of laparoscopic simulation training on improving laparoscopic skills of gynecologic residents with different seniority
Huamao LIANG ; Yiting WANG ; Yan WANG ; Ying YAO ; Yanjie WANG ; Yuan LI ; Kun ZHANG ; Jie QIAO
Chinese Journal of Medical Education Research 2019;18(2):186-190
Objective To explore the value of primary and Advanced levels of laparoscopic simulating training in different seniority of gynecologic residents.Methods 77 residents in their first to forth training-year were divided into two groups:1-2 year resident and 3-4 year resident,trained with different levels of simulating training plans respectively and then assessed in the Department of Obstetrics and Gynecology in Peking University Third Hospital.Results The qualified rate of primary and advanced simulation training was 85.7% and 57.1% respectively.The qualified rate of primary training (80.4% vs.100.0%,P=0.028) and advanced training (12.5% vs.57.14%,P=0.000) were significantly different between 1-2 year residents and 3-4 year residents.The operative skills improved significantly in all the residents.In the 1-2 year residents,the scores of the primary training increased more obviously,while in the 3-4 year residents,the scores of the advanced training increased significantly.Conclusion It might be more effective for residents with different seniority to receive different levels of simulating training accordingly,so as to improve their laparoscopic operative skills more effectively.
6.Application of IQQA three-dimensional reconstruction technique in precise laparoscopic or robot-assisted laparoscopic partial nephrectomy for renal hilar tumors
Yue YANG ; Zhi CAO ; Chao ZHANG ; Huamao YE ; Fei GUO ; Huiqing WANG ; Chen LYU ; Yang WANG ; Bo YANG ; Chuanliang XU ; Yinghao SUN
Chinese Journal of Urology 2017;38(7):493-497
Objective To evaluate the feasibility and effectiveness of the application of IQQA (Intelligent/interactive Qualitative and Quantitative Analysis) three-dimensional reconstruction technique in precise laparoscopic or robot-assisted laparoscopic partial nephrectomy (LPN or RAPN) for renal hilar tumors.Methods The study retrospectively reviewed 11patients with hilar tunors from February 2016 to February 2017.Of the 11 patients,4 were women and 7 were men,with an average age of 51 years (range 38 to 70 years).The average tumor size was 3.1 cm (range 1.7 to 4.3 cm).For tumor stage,9 patients were in T1a stage and 2 patients were in T1b stage.Tbe average R.E.N.A.L score was 8.7 (range 7 to 10).The mean preoperative GFR was 40.6 ml/min (range 32 to 45 ml/min).IQQA three-dimensional reconstruction technique was applied for the purpose of precise navigation and resection of the tumors.Multivariate analysis was used to identify predictors of warm ischemia time,estimated blood loss,major perioperative complications,and postoperative renal function.Results All 11 laparoscopic or robot-assisted laparoscopic hilar partial nephrectomies were successfully completed without conversion to a hand-assisted or an open approach.Under the navigation of IQQA,all tumors were found precisely at the first time during surgeries.The final pathologic examination revealed that all the 11 patients were clear cell renal cell carcinomas.The mean operative time was 142 minutes (range 90 to 230 minutes),with a mean warm ischemia time of 24 minutes (range 17 to 33 minutes).The estinated blood loss was 146 ml (range 50 to 400 ml).No intraoperative complications occurred.Two patients suffered from postoperative complications.One patient with gross hematuria was recovered by consistent bladder irrigation.The other patient with postoperative hemorrhage needed transfusion.All patients had negative margins on the final pathologic examination.At a mean follow-up period of 3 months,the mean GRR is 22.5 ml/min (range 13 to 34 ml/min) without any disease recurrence.Conclusions With peculiar features,such as accurate location,complete resection and fewer perioperative complications,the application of IQQA three-dimensional reconstruction technique in precise partial nephrectomy represents a safe and effective procedure for hilar tumors.
7.Robotic-assisted laparoscopic prostatectomy in patients with high-risk prostate cancer :experiences of 400 cases
Min QU ; Hengzhi LIN ; Haifeng WANG ; Linhui WANG ; Bo YANG ; Huamao YE ; Shancheng REN ; Tie ZHOU ; Chuanliang XU ; Yinghao SUN ; Xu GAO
Chinese Journal of Urology 2017;38(6):424-427
Objective To study the short-term effectiveness of robotic-assisted laparoscopic radical prostatectomy in high-risk prostate cancer.Methods From March 2012 to March 2017,400 patients with high-risk prostate who underwent robotic-assisted laparoscopic radical prostatectomy were reviewed.The median age was 68 years old(ranged from 49 to 83 years),and the median PSA was 23.1 ng/ ml(ranged from 5.2 to 999.0 ng/ml).Preoperative parameters,surgical interventional data,postoperative pathology and follow-up data were collected.Logistic regression was used to analyze the risk factors of positive surgical margin in postoperative pathology.Results All the operations were successfully completed.Median operation time was 115 min(ranged 50-555 min),and median estimated blood loss was 110 ml(ranged 30-500 ml).Six patients had perioperative complications,among which two were rectal injury,two were cardio-cerebrovascular disease and two were hemorrhage.There was no perioperative death.Positive surgical margin was detected in 151 patients,accounting for 37.8%.A total of 345 cases (86.3%) underwent lymphadenectomy,of which 253 cases (63.3%) were performed standardized resection and 92 cases (23%) were performed extensive resection.The median number of resected lymph nodes was 9 (ranged 3-36).Eighty cases (23.2%,80/345) were positive in resected lymph nodes.Regression analysis showed that preoperative PSA > 20 ng / ml or clinical stage ≥ T2c were risk factors for positive surgical margins.After a median follow-up of 14.4 months (ranged 2.0-58.8 months),the overall incidence of biochemical recurrence was 33.4% (107/320),and the urinary continence rate one year after operation was 86.6% (277/320).Conclusions Robotic-assisted laparoscopic radical prostatectomy in patients with high-risk prostate cancer was a feasible,safe and effective approach.Preoperative PSA and clinical stage were the risk factors for positive surgical margin.
8.Surgical outcomes of robot-assisted partial nephrectomy in 12 cases
Linhui WANG ; Huamao YE ; Bin XU ; Bing LIU ; Jiatao JI ; Liang XIAO ; Xia SHENG ; Lili WANG ; Xin CHENG ; Jiong HOU ; Jianguo HOU ; Yinghao SUN
Chinese Journal of Urology 2012;(11):814-817
Objective To summarize the surgical experience in robotic-assisted laparoscopic partial nephrectomy,and to investigate the efficacy and safety of this surgery.Methods The clinical data of 12 patients who underwent robot-assisted laparoscopic partial nephrectomy in Changhai Hospital from March to July in 2012 were analyzed.All the patients were male and the age range was 43-66 years.In 4 cases the tumors were in the left kidney,and 8 in the right.In 7 cases the tumors were in the dorsal part of the kidney,and 2 in the ventral part.There were 3,5 and 4 cases in the upper,middle and lower pole of the kidney respectively.Preoperative GFR test was normal in all cases.Kidney CT scan showed the maximum diameters of the tumors were 2.0-5.8 cm,with an average of 3.3 cm.The pre-operative stages in all cases were T1N0M0.Results The surgery was successfully completed in all cases.The mean duration of the surgery was 160-310 min,with an average of 242 min.The blood loss was 30-300 ml,with an average of 135 ml,and the intraoperative blood transfusion was unnecessary.The warm ischemia time was 20-49 min,with an average of 31 min.There was no intraoperative morbidity,and no conversion to open surgery.The postoperative length of hospitalization was 9-31 d,with an average of 14 d.Gross hematuria arose in 1 patient at 1 week after the surgery.The post-operative pathology showed renal clear cell carcinoma with Furhman Grade Ⅱ in 11 cases,and renal angiomyolipoma in 1 case.The maximum diameters of the tumors were 2.0-5.0 cm,with an average of 3.5 cm.The tumor resection margin was negative in all cases.Conclusions Robot-assisted laparoscopic partial nephrectomy is safe and effective for local renal tumors.This surgery has significant advantage over traditional laparoscopic partial nephrectomy,in terms of the resection of the renal tumors and the reconstruction of the kidney.
9.Application of flipped classroom combined with surgical simulation training system in clinical clerkship teaching of gynecology and obstetrics
Chunyu ZHANG ; Huamao LIANG ; Xuan HE ; Ying SONG ; Ying YAO ; Yan WANG
Chinese Journal of Medical Education Research 2020;19(11):1310-1313
Various contents, relatively insufficient class hours, few practical operation opportunities and lack of teacher-student interaction are the main difficulties in clinical clerkship teaching of gynecology and obstetrics. In this study, the flipped classroom, surgery simulation training system and online self-test module are added to the clinical clerkship teaching of obstetrics and gynecology, and also the WeChat group interactive platform is established to make up for the deficiency of traditional teaching mode. The questionnaire survey showed that 59 clerkship students' satisfaction with the course was 92.6 points, 98.3% of students thought that the surgery simulation training was helpful, and 93.2% of them thought that the flipped classroom was helpful, and the students' scores of the final exam was higher than those of the previous students in the same period with the same difficulty of the exam. Therefore, rational use of flipped classroom, surgery simulation training system and other modules can improve the overall effect of clinical clerkship teaching in gynecology and obstetrics to a certain extent.
10.Safety and prognostic analysis of partial nephrectomy for cT 1N 0M 0 non-clear renal cell carcinoma with high grade malignancy
Jiaao SONG ; Wenqiang LIU ; Bo YANG ; Huamao YE ; Jianguo HOU ; Zhenjie WU ; Linhui WANG
Chinese Journal of Urology 2023;44(6):422-426
Objective:To investigate the safety and prognosis of partial nephrectomy (PN) in the treatment of highly malignant non-clear renal cell carcinoma (nccRCC).Methods:Clinical data of 47 patients with cT 1N 0M 0 high malignant nccRCC treated in Changhai Hospital from March 2016 to March 2022 were retrospectively analyzed. All patients received PN. There were 34(72.3%) males and 13(27.7%) females. The mean age was (53.5±15.0) years, and average BMI, was(23.7±3.4)kg/m 2.The maximum tumor diameter was (29.8±12.6) mm, and R. E.N.A.L. score was 7(5-9), with 37(78.7%) cases of T 1a and 10(21.3%) cases of T 1b. The mean estimated glomerular filtration rate (eGFR) before surgery was (96.3±25.5) ml/ (min·1.73m 2). All patients underwent PN, including 1 patient (2.1%) undergoing open surgery, 29 patients (61.7%) undergoing laparoscopic surgery, and 17 patients (36.2%) undergoing robotic surgery. There were a total of 22(46.8%) cases of papillary cell carcinoma(pRCC)type Ⅱ, 4(8.5%) cases of collecting duct carcinoma (cdRCC), 9(19.1%) cases of MiT family translocated renal cell carcinoma (tRCC), 5(10.6%) cases of mucoid tubular and spindle cell carcinoma (mtSCC)and 7(14.9%) cases of unclassified renal cell carcinoma (uRCC). The surgical conversion rate, positive margin rate, operative time, intraoperative blood loss, complications, and postoperative hospital stay were analyzed. Preoperative and postoperative eGFR were analyzed, and overall survival (OS) and cancer specific survival (CSS) were calculated. Results:All the operations were successfully completed. No radical operation or open operation was performed, with operation time of(100±60) min and intraoperative blood loss of(100±59) ml. There were no intraoperative complication and 1 case (2.1%) suffered from postoperative complication. Postoperative hospital stay were 5 (4-6) days. The mean eGFR after surgery was (86.5±27.1) ml/(min·1.73m 2), and the difference was statistically significant ( P=0.041). In this study, the mean follow-up time was (45.7±20.9)months, and no adjuvant therapy was used after surgery. During the follow-up period, 2 patients died, who all of them were kidney cancer-related death, and both OS and CSS were 95.7% (45/47). Conclusions:PN is safe, feasible and has a good prognosis in the treatment of high malignant T 1 nccRCC. For tumors with clear imaging boundaries and complete envelope, complete tumor resection is more likely, postoperative follow-up should be strict, and no remedial radical or systemic treatment was required.