Objective:To explore the application and preliminary evaluation of multidisciplinary rounds (MDRs) in the clinical teaching of urology.Methods:A total of 20 pediatrics medical students in the clinical medicine were selected as the control group, and the traditional single department teaching rounds were used. Another 20 clinical medical students in the same grade were taken as the experimental group, and MDRs were used. The teaching officer of urology served as the general ward round teacher, combined with nephrology physicians and imaging ultrasound physicians to conduct bedside teaching rounds, and the length of the rounds was about 60 to 90 minutes. Through the teaching evaluation form and the test scores, the effect of teaching ward rounds was evaluated. SPSS 21.0 was used for statistical evaluation data, and the unpaired t-test was performed to make comparison between groups. Results:In terms of theoretical test, the average score of students in the experimental group was (92.15±0.60), which was higher than that in the control group (90.05±0.71), and the difference was statistically significant ( P=0.030). In the experimental group, 95.0%(19/20) of the medical students affirmed the MDRs model, and 100% of them thought it was necessary to introduce this model in clinical teaching. Conclusion:MDRs are patient-centered, emphasize interdisciplinary cooperation, and are operable, which deepens the understanding of medical students on urological diseases, promotes the exchange of clinical teaching experience between urology and interdisciplinary research, and improves the quality of teaching.

Objective:To explore the application value of modified technique of ureter implantation in murine renal transplantation.Methods:Thirty left donor kidneys from BALB/c mice was transplanted into syngeneic mice. Cuff technique was applied for anastomosing kidney artery and vein. The procedure of ureter-bladder anastomoses shifted from implication-fixation-embedding to fixation-implication-embedding. Operative duration, recipient survival rate and complications were recorded.Results:Time for separating vessels, perfusion and excision of donor graft was (25±3) min, (10±6) s for warm ischemia and (25±5) min for cold ischemia. Time for separating recipient vessels was (12±5) min, (7±1) min for arterial anastomosis, (7±1) min for venous anastomosis, (13±2) min for ureter-bladder anastomosis, (5±1) min for right kidney excision and (5±1) min for abdominal closure. Operative duration was(77±3)min. Twenty-six recipients survived over 3 months. The successful operative rate was 86.7%.Conclusions:With a shorter learning curve, modified technique of ureter implantation is easier and faster so as to reduce the postoperative incidence of urinary tract complications during murine renal transplantation.

Objective Establish an overall performance evaluation index system for innovative medical devices transforming project,a major science and technology project in Zhejiang province.Methods The weight of indexes was calculated using Delphi method,analytic hierarchy process and weight of percentile method.Results A three-level performance evaluation index system was established.First level included 3 indexes,as condition index,process index and result index,with weight of 0.3401、0.4042 and 0.2557 respectively.Meanwhile,second level and third level included 13 and 38 indexes.Conclusions The index system provide a performance evaluation tool for the innovative medical devices transforming project.

Objective To analyze the virulence genes and molecular typing of non-O1/non-O139 Vibio cholerae in bloodstream infection,and to provide the scientific basis for its diagnosis,treatment, prevention and controls.Methods Five Vibio cholerae strains were obtained from blood samples of five inpatients with sepsis in Ruian People ’s Hospital from 2012 to 2015 . Morphological examination, biochemical identification,drug sensitivity test and multilocus sequence typing (MLST)classification analysis of strains were conducted.Totally 17 virulence genes were detected by PCR amplification.Results These five suspected Vibrio cholerae isolates were confirmed as non-O1/non-O139 Vibrio cholerae . Drug susceptibility test showed that all the strains were sensitive to tetracycline,ciprofloxacin,piperacillin and tazobactam, meropenem, amikacin and gentamicin; one strain was resistant to trimethoprim/sulfamethoxazole;all were resistant to ampicillin.MLST analysis showed that all strains were new sequence types (ST),belonging to ST268,ST269,ST267,ST270 and ST271 ,and two novel alleles of RY03(mdh:60 and pyrC:86)were discovered.Virulence genes testing showed that the five strains were divided into 4 virulence genotypes:RY02 and RY04 (hlyA + toxR + hap + rtxA + nanH + vasH + vasA +vasK + ),RY01 (hlyA +toxR +hap +rtxA +nanH +vasH -vasA +vasK - ),RY03 (hlyA +toxR +hap +rtxA +nanH - vasH + vasA + vasK + ) and RY05 (hlyA + toxR + hap + rtxA + nanH + vasH - vasA - vasK - ). Conclusions Non-O1/non-O139 Vibrio cholerae can cause human bloodstream infection in immunocompromised patients.The pathogenic factors may be related to the virulence genes of hlyA, toxR,hap ,rtxA and T6SS.

Background and purpose:Cancer of unknown primary (CUP) represents approximately 5%~10%of malignant neoplasms. For CUP patients, identiifcation of tumor origin allows for more speciifc therapeutic regimens and improves outcomes.Methods:By retrieving the gene expression data from ArrayExpress and Gene Expression Omnibus data repositories, we established a comprehensive gene expression database of 5 800 tumor samples encom-passing 22 main tumor types. The support vector machine-recursive feature elimination algorithm was used for feature selection and classiifcation modelling. We further optimized the RNA isolation and real-time quantitative polymerase chain reaction (RTQ-PCR) methods for candidate gene expression proifling and applied the RTQ-PCR assays to a set of formalin-fixed, paraffin-embedded tumor samples.Results:Based on the pan-cancer transcriptome database, we identiifed a list of 96-tumor speciifc genes, including common tumor markers, such as cadherin 1 (CDH1), kallikrein-re-lated peptidase 3 (KLK3), and epidermal growth factor receptor (EGFR). Furthermore, we successfully translated the microarray-based gene expression signature to the RTQ-PCR assays, which allowed an overall success rate of 88.4% (95%CI: 83.2%-92.4%) in classifying 22 different tumor types of 206 formalin-fixed, paraffin-embedded samples. Conclusion:The 96-gene RTQ-PCR assay represents a useful tool for accurately identifying tumor origins. The assay uses RTQ-PCR and routine formalin-ifxed, paraffn-embedded samples, making it suitable for rapid clinical adoption.

Objective To study the effects of using 25 kDa linear and branched PEI to transfer plasmid DNA pEGFP -C1 (pDNA ,encoding the enhanced green fluorescent protein reporter gene ) to the basilar membrane of the C57BL/6 mice cochlea in vitro .Methods L -PEI/pDNA and B -PEI/pDNA polyplexes were generated in 0 .1M phosphate buffer solution (PBS) or 5% glucose solution .Polyplexes were characterized by transmission electron mi-croscopy .Agarose gel retardation assay was used to determine the plasmid binding ability of L -PEI and B -PEI . The toxicity was investigated by MTT assay .The transfection was firstly evaluated in 293T cell line ,and then the appropriate amount of PEI and plasmid were applied for cochlear explant transfection of P4 mice pups .Results Un-der the same condition ,B -PEI had better transfection efficiency than L -PEI ,but its toxicity was also higher . When generated in PBS ,the polyplexes had lower toxicity than in glucose solution .L -PEI-pDNA nanoparticles could transfect the spiral limbus fibrocytes ,some spiral ganglion neurons and supporting cells ,but the efficiency was low .Conclusion L -PEI could be used as the non -viral vector for the transfection of the cultured basilar mem-brane of P4 mice pups ,but it should be modified to reach higher efficiency .

Objective To investigate the phenotype and function of antigen-specific CD8+ cytotoxic T cells (CTL) from HLA-A2+ healthy human donors induced by high-carcinogenic HPV 16 E7 peptide in vitro.Methods Peripheral blood T cells from 26 HLA-A2+ healthy human donors were incubated with HPV 16E711-20 peptide (HLA-A*0201/YMLDLQPETT), recombinant human interleukin 7 (IL-7) and IL-2 for 7 days to yield antigen-specific CD8+ CTL. Then, four-color flow cytometric analysis was performed to detect the percentage of antigen-specific CD8+ CTL expressing different surface markers including CD45 and CD27. The expression of three intracellular cytokines including perforin, granzyme-B and FasL in the antigen-specific CD8+ CTL was also measured by intracellular flow cytometry. A wrong peptide, HBVcoxe18-27 (FLPSDFFPSV),was used as an isotype control. Results A significant increase was observed in the percentage of antigen-specific CD8+ CTL in peripheral T cells stimulated with HPV 16 E7-peptide compared with the non-stimulated T cells (0.73% ± 0.33% vs 0.02% ± 0.03%, P ＜ 0.01). The percentage of CD45RA+CD27- effector T cells,CD45RA-CD27- effector memory T (TEM) cells, CD45RA-CD27+ central memory T (TCM) cells, and CD45RA+CD27+ naive T cells in antigen-specific CD8+ CTL was 26.07% ± 13.46%, 7.97% ± 7.11%, 33.25% ± 19.68%and 32.73% ± 13.89%, respectively in HPV 16 E7-stimulated group, significantly higher than that in nonstimulated group (0.02% ± 0.03%, 0.02% ± 0.03%, 0.02% ± 0.03% and 0.02% ± 0.05%, all P ＜ 0.01 ). Elevated proportions of perforin-, granzyme-B- and FasL-expressing antigen specific CTL were observed in HPV 16 E7-stimulated group compared with non-stimulated group (47.01% ± 18.69% vs 0.38% ± 0.55%, 80.53% ±13.32% vs 0.34% ± 0.22%, 26.48% ± 7.81% vs 0.16% ± 0.16%, all P ＜ 0.01 ). Conclusions HPV 16 E7peptide could induce the clonal proliferation of CD8+ T cells, generation of antigen-specific CD8+ CTL and secretion of toxic cytokines, finally lead to a highly efficient and specific killing of virus-infected target cells through different mechanisms, hence, it might play a crucial role in antiviral immune responses.

Objective To study the effect of acute hypervolemic hemodilution on expression of serum chemokine interferon-inducible protein 10 in patients undergoing total hip replacement.Methods Twenty ASA Ⅰ or Ⅱ patients undergoing elective total hip replacement were randomly divided into 2 groups (n=10 each):HES group and LR group.The patients in HES group received 6% HES 20 ml/kg in rate of 30 ml/(kg·h) after anesthesia.The patients in LR group received Ringer's solution 20 ml/kg in rate of 30 ml(kg·h) after anesthesia.The blood loss,blood transfusion and the time of operation were recorded.Venous blood samples were taken before anesthesia (T0),at the begining of operation (T1),30 min after operation (T2),and at the end of operation (T3),in determination of serum chemokine interferon-inducible protein 10.Results The blood loss and the blood transfusion in HES group were (560±90)ml and (200±100) ml,those were significantly lower than that in LR group[(810±110)ml and (600±200)ml].The IP-10 concentrations were significantly increased at T2～T3 as compared to baseline value at T0 in both groups,but were higher in LP group[(77.3±13.8) ng/L and (89.9±15.1) ng/L]than those in HES group [(62.8±13.6) ng/L and (65.4±10.2) ng/L,P＜0.05].Conclusions Acute hypervolemic hemodilution can abate blood loss and blood transfusion during total hip replacement operation.Preoprative infusion with hydroxyethyl starch can attenuate the immunological depression during operation and anesthesia.

Objective To study the effect of acute hypervolemic hemodilution on expression of plasma bac-tericidaL/permeability-increasing protein (BPI) in patients undergoing total hip replacement. Methods Twenty ASA Ⅰ-Ⅱ patients undergoing elective total hip replacement were randomly divided into two groups (n=10 for thesia. The blood loss,blood transfusion and the time of operation were recorded. Venous blood samples were taken before anesthesia (T0) ,at the begining of operation (T1) ,30 min after operation (T2) ,and at the end of operation (T3) for determination of plasma bactericidal/permeability-increasing protein. Results The blood loss and the blood transfusion in HES group were significantly lower than that of LR group[blood loss: (560±90)ml vs (810±110) ml and blood transfusion: (200±100) ml vs (600±200) ml,t=5.562 and 5.657,P<0.001]. The plasma BPI concentrations in HES group were significantly increased at T2～T3 as compared to baseline value at T0 [(8.9±1.6)μg/L,(13.4±1.2)μg/L and (4.9±1.2)μg/L,P<0.05]. The plasma BPI concentrations in LR group were significantly increased at T2～T3 as compared to baseline value at T0 [(7.3±1.2)μg/L,(9.9±0.8) μg/L and (5.0±1.1)μg/L,P<0.05],but were lower than those in HES group (t=2.530 and 7.674,P=0.021 and 0.001 ). Conclusion Acute hypervolemic hemodilution with 200/0.5 hydroxyethyl starch can reduce blood transfusion during total hip replacement operation and also can increase the BPI level which would beneficial for the immunological function.

Objective To investigate the possible roles of cellular immunosuppression induced by phenotypic and functional changes of peripheral CD4+CD25+ regulatory T cells (Tregs) in the pathogenesis of condylomata acuminata. Methods Three-color flow cytometry was performed to examine the expression of transcription factor Foxp3 in, along with several inhibitory membrane molecules, i.e. cytotoxic T lympho-cyte associated antigen-4 (CTLA-4), glucocorticoid-induced TNF receptor family-related gene (GITR) and programmed death-1 (PD-1) on peripheral CD4+CD25+ T cells from 46 patients with condylomata acuminata and 43 normal human controls. Meanwhile, high purity of CD4+CD25+ T cells were isolated from peripheral blood using imrnunomagnetic beads, and stimulated to produce intracellular suppressor cytokines such as interleukin-10 (IL-10) and transforming growth factor-beta (TGF-beta), which were detected by flow tyro-metric analysis. Results The number of peripheral CD4+CD25+ Foxp3+ Tregs increased significantly in patients with condylomata acuminata than that in the normal controls (7.37% ± 2.43% vs 5.96% ± 2.09%,P ＜ 0.001). The expressions of CTLA-4 and PD-1 were 1.86% ± 1.13% and 2.41% ± 1.12%, respectively,in the patients, which were significantly higher than those in the normal controls(1.36% ± 0.90% and 1.70%± 0.97%, P ＜ 0.05 and 0.01, respectively). The number of TGF-beta-positive CD4+CD25+ T ceils from peripheral blood were increased in patients than that in, the controls (1.57% ± 0.91% vs 0.78% ± 0.24%, P ＜0.001). Conclusions Human papillomavirus infection can induce the activation and proliferation of CD4+CD25+ Tregs, enhance the expression of negative costimulatory molecules and secretion of suppressor cytokines, and inhibit antiviral immune response through multiple mechanisms.