Cytological study on aptamer selection for bladder cancer through CELL-SELEX / 医学研究生学报

ObjectiveThe methods based on bladder cancer markers which could be applied to early diagnosis and postoperative recurrence monitoring of bladder cancer were current research hotspots. This study aims to screen aptamers that specifically recognize human bladder cancer cell lines (EJ, T24, BIU87) through cell-based systematic evolution of ligand by exponential enrichment (CELL-SELEX).MethodsFor CELL-SELEX screening, bladder cancer cell lines EJ, T24, and BIU87 were used as positive control cells. HCV 29 (human normal urothelial cell line), 293T (human embryonic kidney cell line), huh7 (human hepatocellular carcinoma cell line) were used as negative control cells. PCR upstream primers were labeled with FITC, downstream primer was labeled with Biotin. ssDNA fragments collected from each round were amplified by PCR, and the amplified product was then purified using a DNA purification Kit. The biotin-streptavidin magnetic separation methods were used to isolate the PCR product to obtain secondary FITC-ssDNA for the next CELL-SELEX round. The screening process was monitored by flow cytometry. ssDNA pool with the highest binding rates to bladder cancer cell lines(EJ, T24, and BIU87) was selected to PCR amplification, product purification, molecular cloning, and sequencing. According to the sequencing results, the secondary structure of the aptamer was pre-simulated by Dnaman software. Aptamer labeled with FITC was synthesized in vitro, flow cytometry was used to detect the binding rate of the aptamer to bladder cancer cell lins (EJ, T24 and BIU87).ResultsWith the advance of the CELL-SELEX process, the binding rate of FITC-ssDNA to bladder cancer cell lins (EJ, T24, and BIU87) increased gradually. By the 15th round, the binding rate of FITC-ssDNA to EJ cells reached the highest level. The apt1 had the highest enrichment among the 15th round ssDNA pool. By the 18th round, the binding rate of FITC-ssDNA to T24 or BIU87 cells reached the highest level. The apt2 and apt3 had the highest enrichment among the 18th round ssDNA pool. DNA structure prediction showed that the secondary structure of apt1, apt2, and apt3 was mainly stem-loop structure. Flow cytometry showed that the highest binding rate was FITC-apt1 to EJ cells, FITC-apt2 to T24 cells, and FITC-apt3 to BIU87 cells, respectively. There is no significant combination between these aptamers with the negative cells.ConclusionIn this study, three kinds of aptamers with high specificity for bladder cancer cell lines were successfully screened by CELL-SELEX. The apt1 can specifically recognize EJ cells, apt2 can specifically recognize T24 cells and apt3 can specifically recognize BIU87 cells, all of which provide experimental evidence for early diagnosis and targeted therapy technology research of bladder cancer.

Research progress of vascular endothelial growth factor in bladder cancer / 医学研究生学报

Vascular endothelial growth factor (VEGF) is known as the most efficient promote blood vessel growth factor.Numerical novel researches has confirmed that it plays a key role in bladder cancer proliferation, invasion and metastasis. The increasing clinical trials have aimed at targeting VEGF pathway.The encouraging outcomes provide new feasibility on bladder tumor treatment. Theis review conducted to discuss the tumor promoting mechanisms, bladder cancer progression relevance and bladder tumor target therapy of VEGF.

A Clinical Analysis of 554 Cases of Partial Tubeless mPCNL / 昆明医科大学学报

Objective To summarize the experience of partial tubeless mPCNL. Methods A retrospective analysis of partial tubeless mPCNL surgery experience from January 2010 to December 2016. Atotal of 1320 patients underwent mPCNL surgery in these 7 years.Out fo those 1320 patients,554 patients underwent partial tubeless mPCNL,766 patients underwent standard mPCNL,and 85 exception cases of standard mPCNL were forced to abort surgery due to maximum surgery time of 2 hours and different complications such as bleeding, infections and etc, so total of 681 patients with standard mPCNL were compared with partial tubeless mPCNL.Results The rate of partialtubeless mPCNL has increased by 84% in 2016, with an indication of tubeless mPCNL being extended, while the complication rate showed no increase. Compared with the standard mPCNL, there was no significant dif-ference between the two groups in the rate of stone removal, drop in mean hemoglobin, blood transfusion and postoperative fever. There were significant differences in postoperative analgesic use rate (5%:21%,P=0.001) and hospitalization stay (2.5:4.5d,P=0.001) . The rate of postoperative bleeding complications in partial tubeless group and standard group is 1.1%and 2.5%respectively, but difference is not statistically significant. There were 1cases of urinary extravasations in the partial tubeless group which was treated by perirenal drainage, antibiotic treatment, and 1 cases of pleural injury, which were treated by open exploration, and chest tube placement. Conclusion In compared to standard mPCNL partial tubeless mPCNL significantly increased patients postoperative comfort, shorten hospital stay,and no complications increased.Partial tubless mPCNL is a safe and practicable procedure.

The Research of Monoclonal Antibody KMP1 Inhibited Bladder Cancer EJ Cell Lines Growth and Metastasis in Vivo / 昆明医科大学学报

Objective To research the monoclonal antibody KMP1 inhibited bladder cancer EJ cell lines growth and metastasis in vivo by bioluminescence imaging. Methods Immunohistochemistry was used to determine the KMP1 binding to EJ and EJ-GFP cell lines. The xenograft tumor cell growth and distribution were measured by vernier calipers and dynamic in vivo fluorescence imaging. Immunohistochemistry and H&E counterstaining researched the feature of the xenograft tumor. Results Cell growth curves of EJ and EJ-GFP cells were similar. EJ-GFP had a green fluorescence. In EJ-GFP nude mouse tumor model, the addition of KMP1 significantly inhibited tumor growth and extended the average life span of nude mice. Both EJ and EJ-GFP cells can bind to KMP1,and the weight of transplanted tumors in the KMP1 treatment group was significantly lower than that of the mIgG control group (P＜0.001).Conclusion KMP1 has a promising antitumor effect in vivo. It might be valuable for development as a promising targeted agent for bladder cancer.

Update of combined therapies with docetaxel for hormone-independent prostate cancer / 中华男科学杂志

Hormone-independent prostate cancer (HIPC) is the end stage of prostate cancer, with a short median survival of 9-18 months for the patients. Two large phase III studies have demonstrated a survival advantage of docetaxel chemotherapy in HIPC patients. New combined protocols have been developed with promising results. These protocols propose a combination with docetaxel, chemotherapy, antiangiogenic agents, vaccine and biological drugs. This review focuses the progress achieved the combined therapies for HIPC.

Characterization of a novel transplantable orthotopic nude mouse model with xenografted human bladder transitional cell tumor (BIU-87) / 中华肿瘤杂志

<p><b>OBJECTIVE</b>A mouse model of orthotopic bladder cancer simulating its human counterpart is of great importance in preclinical evaluation of new treatment modalities such as immunotxin therapy. The aim of the present study is to establish a novel nude mouse model with xenografted human bladder cancer.</p><p><b>METHODS</b>Single cell suspension of an established human bladder transitional cell carcinoma (TCC) cell line BIU-87 was instilled into nude mouse bladders which were pretreated with mild acid washing. The tumor growth in mouse bladder was assessed weekly by magnetic resonance imaging (MRI). At intervals following implantation and MRI tumor detection, the animals were sacrificed for necropsy, histological examination and immunocytochemical studies.</p><p><b>RESULTS</b>The overall tumor establishment was 92.9% (52/56 mice) at 7 - 36 days, while in the subgroup of animals sacrificed at 12 - 13 days, 40 out of 42 animals (95.2%) developed TCC, the majority of which was superficial. The tumor stages were assessed by gross and histopathology. Histological examination confirmed the presence of grade II - III TCC. Immunocytochemistry confirmed that the tumor model maintained the biological and immunological features of BIU-87 cells. The changes seen on MRI images well correlated with the extent of tumor invasion identified by histology. Carcinoma in situ could be detected histologically at 7 - 9 days post-inoculation and progressed into papillary or invasive tumors thereafter.</p><p><b>CONCLUSION</b>The orthotopic BIU-87 TCC model in nude mice is highly reproducible and is ideal for preclinical studies on experimental intravesical therapies.</p>

Preparation of lung targeting azithromycin liposomes and its tissue distribution in mice / 药学学报

<p><b>AIM</b>To prepare lung targeting azithromycin cationic liposomes and to observe its tissue distribution in mice.</p><p><b>METHODS</b>The azithromycin cationic liposomes were prepared by thin film method with freeze-thawing steps. HPLC method was established and validated for the determination of azithromycin in tissues of mice.</p><p><b>RESULTS</b>The particle size of the liposomes was 6.582 microm with zeta potential of +19.5 mV. The entrapment efficiency was more than 75%. The liposomes was stable in 6 months stored at 4 degrees C. The release in vitro was characterized by Higuchi equation. Azithromycin liposomes and free azithromycin solution were injected intravenously at a dose of 80 mg x kg(-1) to mice. Compared with solution, liposomes were characterized by slower clearance, increased half-life and the AUC increased by 7.4 fold in lung.</p><p><b>CONCLUSION</b>Thin film method with freeze-thawing steps could increase the entrapment efficiency and increase the particle size of azithromycin liposomes. After modification of lipid membrane with stearylamine, the cationic liposomes were prepared. The azithromycin concentration and AUC increased in lung after iv administration to mice of the cationic liposomes. This offered a good information for preparing liposomes targeting on the lung.</p>

Comparison of clinical efficacy of orthotopic ileal neobladder versus orthotopic sigmoid neobladder / 中华泌尿外科杂志

Objective To compare the clinical efficacy of orthotopic ileal neobladder versus ortho- topic sigmoid neobladder.Methods The data of 96 patients who had undergone orthotopic ileal neoblad- der and 68 patients who had undergone orthotopic sigmoid neobladder were retrospectively analyzed.The perioperative condition,urinary continence,urodynamics,and pouch-related complications were compared between the 2 groups.Results Of all the 164 patients,12(7.3%)were lost to follow-up.The mean fol- low-up was 46(2-86)months in orthotopic ileal neobladder group,and 42(4-78)months in orthotopic sigmoid neobladder group.There was no significant difference in intraoperative blood loss and postoperative urinary continence between the 2 approaches(P＞0.05).However,compared with sigmoid neobladder group,ileal neobladder group had longer operative time and postoperative recovery time,and got a bigger pouch(P＜0.05).The early and late pouch-related complication rates of ileal neohladder group were 16. 7% and 29.2%,which were higher than those of sigmoid neobladder group.During the follow-up,tumor recurred in 3 cases of ileal neobladder group,but none in sigmoid neobladder group.Conclusions Ortho- topic ileal neobladder and sigmoid neobladder are similar in operative difficulties,and both can achieve satis- factory clinical results.Compared with ileal neobladder,sigmoid neobladder has shorter operative time, quicker recovery and lower rate of pouch-related complications,thus is a preferred procedure.