Comparative study of fluorescence vs.high-definition laparoscopy in extended pelvic lymph node dissection plus radical prostatectomy for patients with locally advanced prostate cancer / 中华泌尿外科杂志

Objective To compare the efficacy of extended pelvic lymph node dissection (ePLND)and oncological outcome by fluorescence laparoscopic radical prostatectomy (FLRP) versus high-definition laparoscopic radical prostatectomy (HD-LRP) for men with locally advanced prostate cancer (LAPCa).Methods In a prospective trial,we recruited 51 patients with T3a-bNxM0 prostate cancer from July 2015 to April 2018.Patients were assigned to study group or control group according to random number method,and were underwent either FLRP + ePLND or HD-LRP + ePLND.21 in the study group were injected with 5 mg of indocyanine green (ICG) into the bilateral lobes of the prostate transperineally guiled by transrectal ultrasound 30 min before surgery for lymphography.During the surgical procedure a fluorescence laparoscope,optimized for detection in the near infrared range,was used to visualize the lymph nodes (green fluorescent) in the dissection region in the study group while a common laparoscopy introduced in control one.Lymph nodes were removed in the external iliac vessiles,internal iliac artery,obturator fossa regions,common iliac regions and presacral regions in both groups.Radical prostatectomy was completed in the both groups by similar steps.The operation time,blood loss,number of removed lymph nodes and positive lymph nodes,complication rate,biochemical recurrence (BCR) and metastasis free survival rates in 2 years were recorded and compared in the two groups.Results 51 eligible patients were selected,including 21 in the study group and 30 in the control group.The mean age of biopsy of study group and control one were (66.4 ± 7.7) and (66.8 ± 7.4),the mean age PSA (23.5 ± 16.8) ng/ml and (26.0 ± 20.1) ng/ml,the mean Gleason score of biopsy (8.1 ± 1.0) and (7.9 ± 0.9) respectively,and there was no statistical significant difference between two groups.The mean operation time of study group and control one were (45.9 ± 4.6) min and (56.4 ± 3.2) min,the mean removed lymph nodes were (27.7 ± 5.6) and (22.1 ±5.6) respectively,and there was statistical significant difference between two groups (all P ＜ 0.05).Lymph nodes invasion in pathology were reported in 8 cases(38.1％)in the study groups while 9 (30.0 ％) in the control one;the proportion of positive lymph node (metastasis) were 3.2％ (19/583) and 3.4％ (23/663) in the two groups respectively and no statistically significant difference was noted between the two groups.Lymphorrhagia occurred in 4 cases in the control group,and there was no serious complications in both groups.The median follow-up time was 20 (7-33) month and during this time,BCR observed of 1 (4.7％) in the study group and 8 (26.7％) in the control;meanwhile,the MFSR was recorded of 100.0％ (0)in the study group and 86.7％ (4)in the control one,showing a statistically significant difference between the two groups(P =0.04).Conclusions Comparing with LRP,FLRP achieved better results of LN dissection,which will improve oncological outcomes.

Value of endonuclease domain containing 1 in progression of prostate cancer / 中国病理生理杂志

AIM: To analyze the difference of endonuclease domain containing 1 (ENDOD1) expression be-tween benign prostatic hyperplasia ( BPH ) tissues and prostate cancer ( PCa ) tissues and to investigate the effect of ENDOD1 on the biological function of human prostate cancer cells .METHODS: The BPH samples ( n=20 ) and PCa samples (n=21) were processed and analyzed according to the instruction of immunohistochemical (IHC) staining.The mRNA and protein levels of ENDOD 1 in the normal prostate epithelial cells and prostate cancer cells were evaluated by RT -qPCR and Western blot , respectively .The recombinant plasmids pCMV-N-Flag-ENDOD1 was constructed and was trans-fected into the human prostate cancer cells .The proliferation , apoptosis , migration and invasion abilities of the prostate cancer cells were evaluated by MTT assay , flow cytometry, Transwell migration and Matrigel invasion assays , respectively. RESULTS:The analysis of variance of the immunoreactivity score showed that PCa tissues with high Gleason score dis -played significantly lower ENDOD1expression than that with low Gleason score and BPH (P<0.05).The expression of ENDOD1 at mRNA and protein levels in PC3 cells and DU145 cells was significantly lower than that in the LNCap cells (P<0.05).The proliferation of DU145 transfected with ENDOD1 was inhibited.The flow cytometry indicated that ENDOD1 over-expression in the DU145 cells resulted in a notable increase in G0/G1 phase arrest (P<0.05), but the ap-optotic rates showed no statistical difference .The results of Transwell assay showed that migration and invasion abilities of the cells were also inhibited after transfection with over-expressing ENDOD1 plasmid (P<0.05).CONCLUSION: The expression of ENDOD1 significantly decreased in prostate cancer with high Gleaon score .Meanwhile, the ENDOD1 is spe-cifically down-regulated in androgen independent prostate cancer (AIPC) cell lines.Over-expression of ENDOD1 remark-ably inhibits the proliferation , migration and invasion abilities of AIPC .

Preparation Technology Optimization of Ketoconazole and Miconazole Nitrate β- Cyclodextrin Inclusion Compound / 中国药师

Objective:To optimize the preparation technology of ketoconazole and miconazole nitrateβ-cyclodextrin inclusion com-pound. Methods: The weight ratio of β-cyclodextrin to ketoconazole, inclusion temperature and inclusion time as the testing factors, the optimal inclusion technology was screened by orthogonal experiments. Results:The optimum inclusion conditions were as follows:the weight ratio of β-cyclodextrin to ketoconazole was 8 ∶1, the inclusion temperature was 50℃, and the ultrasonic time was 50 min. Conclusion:The optimized β-cyclodextrin inclusion process is simple and convenient to carry out.

The efficacy of calcined natural bovine bone(CBB)in the treatment of bone defects in animal / 实用口腔医学杂志

Objective:To observe the efficacy of calcined bovine bone(CBB)in the repair of animal bone defects.Methods:Ca-nine alveolar bone defect model,rat and rabbit calvarial critical size defect models were established respectively.All animals were randomly divided into 2 groups(n=6).The defects in the experimental group were repaire with CBB,those in the control group were not treated.The effects were observed by HE staining,Micro CT and Masson trichrome staining.Results:Canin alveolar bone de-fects were well repaired 8 week after operation.The skull defects in rabbits and SD rats were replaced by new bone 1 2 and 8 weeks after operation respectively.In all the control groups bone defects were not healed or not completely repaired.Conclusion:CBB is ef-fective in the repair of bone defects.

Simultaneous Determination of Ketoconazole and Miconazole Mitrate in Compound Toconazole Gels by HPLC / 中国药师

Objective: To establish a method for the determination of ketoconazole and miconazole nitrate in compound ketocon-azole gels. Methods:An HPLC method was developed. A Hypersil BDS C18 column(416 mm × 200 mm,5μm) was used, the mobile phases consisted of 0. 5% ammonium acetate solution-methanol(containing 0. 2% triethanolamine) (20∶80), the flow rate was 1. 0 ml ·min-1 , the detection wavelength was set at 230nm, the column temperature was 30℃ and the injection volume was 20μl. Results:There was a good linear correlation within the range of 5. 1-510. 0 mg·L-1 for ketoconazole (r=0. 999 9) and 50. 0-500. 0 mg·L-1 for miconazole nitrate (r=0. 999 9), the average recovery for ketoconazole and miconazole nitrate was 100. 3%(RSD=0. 38%, n=6) and 99. 9%(RSD=0. 79%, n=6), respectively. Conclusion:The method is simple, rapid, accurate and sensitive, and can pro-vide a method for controlling the quality of compound ketoconazole gels.

Effect of silencing human RRM 2 by specific siRNA on the biological behavior of human osteosarcoma cell / 中国基层医药

Objective To investigate the effect of small interference RNA(siRNA) targeting RRM2 on the biological behavior of human osteosarcoma cell line Saos-2 and the molecular mechanisms.Methods RRM2 expression was knocked down in human osteosarcama cell line Saos-2 by RRM2 siRNA.The expression of RRM2 mRNA and protein was determined in human osteosarcoma cell line Saos-2 and human osteoblast-like cell line hFOB1.19 by real time-PCR and Western blot.The cell proliferation was detected by CCK-8.The migration was observed by using transwell system.The apoptotic rate was observed by ELISA.The expression of CyclinD1 and Bcl-2 proteins were detected by Western blot.Results The expression of RRM2 mRNA and protein was higher in Saos-2 than in hFOB1.19.siRRM2 could down-regulate the expression of RRM2 in Saos-2 cells in a time-and concentration-dependent manner.CCK-8 assay showed that si-RRM2 could inhibit the proliferation ability of Saos-2 cells in a time-and concentrationdependent manner,but had no effect on the proliferation of hFOB1.19 cells.Transwell assay indicated that si-RRM2 could inhibit the migration of Saos-2 cells.si-RRM2 combined with adriamycin could increase the apoptosis of Saos-2 cells.Western blot showed that the expression of Cyclin D1 and Bcl-2 were decreased by silencing RRM2.Condusion RRM2 overexpression maybe associate with the osteosarcoma cells proliferation and migration and suppression of its function is a potential therapeutic strategy in osteosarcoma.

Transvesical approach laparoendoscopic single-site radical prostatectomy: for organ-confined prostate cancer: report of 8 cases / 中华泌尿外科杂志

Objective　To investigate the feasibility of applying transvescal approach laparoendoscopic single-site radical prostatectomy (TVSSLRP) and assess the oncological and functional outcomes.Methods　Eight patients with clinically localized prostate cancer (PCa) of low risk underwent TVSSLRP.Demographic data were accrued including patient age,body mass index (BMI),preoperative PSA level,the International Index of Erectile Function 5,biopsy Gleason score,clinical TNM stage and D'Amico risk classification.One surgeon performed all TVSSLRP procedures.A homemade triple-port was introduced percutaneouly into the bladder to establish pneumovesicum through a 4 cm incision.The major steps of the surgery were described as follows:initial incision was made along posterior margin of the bladder neck to expose bilateral vas deference and spermatic vesicle.After opening Denonvilliers' fascia and extending the space to lateral prostatic pedicles,an intra-fascial nerve sparing procedure was performed.The puboprastatic ligaments were then separated close to the prostate surface and the dorsal vein complex was cautiously swept off.Subsequently,careful apical dissection and urethral transection was sequentially conducted. To reduce the tension of vesico-urethral anastomosis,3 additional incisions parallel to vesio-urethral margin were created and a novel tension - reduced V-LocTM barbed polydioxanone sutures was used.　Results　All the operations were successfully performed and there was no conversion to standard laparoscopic approach or open surgery.The total operative time range was 75 - 180 min with mean time of 125 min.The blood loss was 85 -450 ml with mean 140 ml and no blood transfusion was required.The catheter was removed after a mean (range) of 14 (9 -16) days.No intra-operative complications occurred. No patient had positive surgical margins.The mean (range) hospital stay was 17 (13 -25) days after surgery. All the cases were continent after removal of the catheter.No cases demonstrated vesico-urethral stricture and biochemical recurrence on 12 - 18 months follow up postoperatively.　Conclusions　TVSSLRP is technically feasible for cases with organ-confined prostate cancer with good oncological and functional results.

The classification of ureter disease by the level of difficulty and risk during rigid ureteroscopic surgery / 中华泌尿外科杂志

Objective To summarize the common types and clinical characteristics of ureter disease;which can increase manipulation difficulties and adverse events during rigid ureteroscopic procedures. Methods From Jan 2001 to Dec 2010,our team performed 317 rigid ureteroscopic Drocedures for ureteroscopic examination or treatment;including 60 difficult procedures(34 male and 26 female).The mean age of the patients was 37 years (range,18 to 71).The ureteral diseases were classifted into five types according to the pathological characteristics:Type Ⅰ calculous stenosis,Type Ⅱ neoplastic stenosis;Type Ⅲ non-congenital stenosis,Type Ⅳ congenital stenosis,Type Ⅴ expansion of tortuous ureters.The operative time,complications,and conversion to open surgery were evaluated,and the therapeutic methods were analyzed. Results Of the 60 difficuhly-manipulated procedures,the mean manipulated time was 75 min (range,31 to 200).Intra-operative complications occurred in 9 procedures,including 4 cases of mucosal bleeding,2 cases of submucosaI false passage and 3 cases of ureteral perforation.Eleven procedures were converted to open surgery. In five procedures only a double J tube was inserted for drainage due to the difficulty of entering the ureter.Fiftyfive patients were followed up for 17 months (range,3 to 110);48 patients were cured,5 patients improved and 2 patients were unchanged. Conclusions The five types of ureteral disease can increase operative difficulties and risks of rigid ureteroscopic procedures.We should be cautious during surgery and should stop manipulation or convert to other surgeries if necessary.

Combined Action of ACE Gene I/D and GNB3 Gene C825T Polymorphisms on Essential Hypertension in Northern Han Chinese / 生物化学与生物物理进展

Essential hypertension (EH), a complex polygenic disease, is considered to the result of the genetic interaction of multiple gene alterations in concert with environmental factors. Evidences showed that angiotensin-converting enzyme (ACE) gene and G protein beta3 subunit (GNB3) gene are both important susceptibility genes for EH, and that there exists putative biological connection between the two genes in developing hypertension. To investigate whether hypertension was affected by gene-gene interaction between the two genes in the northern Chinese Han population, a case-control association study including 502 hypertensive cases and 490healthy controls was conducted, selecting the ACE gene I/D polymorpinsm and the GNB3 gene C825T polymorphism. Linkage disequilibrium analysis revealed a significant nonrandom distribution only in male hypertensives, indicating that interaction between ACE gene and GNB3 gene may predispose males to the occurrence of hypertension. Multivariate stepwise logistic regression in single locus analysis, with adjustment for common risk factors for hypertension, demonstrated that the OR for DD/ID versus Ⅱ for hypertension among men was significant (OR 1.57; 95% CI, 1.09 ～2.27; P = 0.016) in dominant genetic model. In combination analysis stratified with respect to gender, slightly significant ORs were found after adjustment in males: OR for TT vs CC, 0.11; 95%CI, 0.01 ～0.99; P = 0.049 within ACE DD genotype; OR for DD/ID vs Ⅱ, 1.52; 95% CI, 1.01 ～2.29; P = 0.047 within GNB3 CC+CT genotype. The results suggest that ACE, or a nearby gene, is a male-specific susceptible gene for hypertension, and that there may exist epistatic gene-gene interaction between ACE D allele and GNB3 825C allele.

Expression of recombinant human tumor suppressor NDPK-A in E. coli / 中国病理生理杂志

AIM: To construct E. coli expression plasmid of recombinant human NDPK-A with a 6?His tag, optimize the expression condition and identify the activity of the product. METHODS: nm23-H1 was subcloned from plasmid pBVNMH1 to pQE40 which contain 6?His purification tag. The expression condition was modulated in grades to get the optimal expression. We purified protein with the Ni+-NTA affinity chromatography column, identified the immunogenicity of the product with Western blot, and measured the kinases activity with HPLC. In addition, angiogenesis inhibition activity of rhNDPK was identified by CAM. RESULTS: The sequence of nm23-H1 subclone in pQE40 was exactly correct. The expression rate of rhNDPK-A was 49 6%. Purified rhNDPK-A specially recognized the antiserum of NDPK-A. It also inhibited angiogenesis. CONCLUSION: PQE-nm23H1 containing 6?His can express target protein at high level. This purification method is simple than other methods, and the product has the same activity as natural human NDPK-A.