To increase the biotransfomation efficiency from the orotic acid to the uridine 5'-monophosphate(UMP),URA5 gene encoding orotate phosphoribosytransferase was amplified from Saccharomyces cerevisiae BY4742 by PCR,then it was inserted into the expression vector pYX212(contained orotidine monophosphate decarboxylase gene URA3)and the pYX212-URA5 was transformed into Saccharomyces cerevisiae BJX12 by electroporation.The recombinant strain was elementarily used to convert orotic acid to UMP.The results showed that pYX212-URA5/BJX12 could accumulate 7mmol/L UMP from 32mmol/L orotic acid in 26h,significantly higher than both control groups pYX212/BJX12(2.7mmol/L) and BJX12(2.4 mmol/L).

Objective To explore the risk factors and metastasis of non-sentinel lymph node (NSLN) in breast cancer patients with sentinel lymph node metastasis≤2. Methods 88 cases of breast cancer with sentinel lymph node metastasis≤2 were retrospectively analyzed. Factors influencing the status of NSLN were studied. Results 8 cases (9.09%) had NSLN metastasis. Variance analysis showed that tumor stage(P=0.014), histolog-ical grade(P=0.014) and ER statuts(P=0.009) were correlated with NSLN metastasis. Logistic analysis showed that only histological grade was the independent predictor for NSLN metastasis (P=0.016,OR=1.150,95%CI=0.598~7.740).18 cases had micro-metastasis of sentinel lymph node and 70 cases had macro-metastasis of sentinel lymph node. The ratio of false negative SLN was 10.23%. The mean follow-up time was 11 months. There was no case of recurrence, metastasis or death. Conclusions The metastasis rate of NSLN is relatively low in breast cancer patients with small number of positive SLNs. NSLN metastasis is related to tumor stage, histological grade, and ER expression.

Objective To study the effect of Yuan‐source Points and Collateral‐points combination and Bushen Yishou cap‐sule on hemorheology and hs‐CRP in patients with cerebral infarction .Methods A total of 120 patients with neurological function defects diagnosed with cerebral brain infarction were randomly divided into 3 groups ,who were given general treatment and nursing as usual ,Acupuncture group in general treatment based on Yuan‐source Points and Collateral‐points combination for enjoy points principles ,to acupuncture treatment ;drug group to oral Bushen Yishou capsule;combined group to Yuan‐source Points and Collat‐eral‐points combination and oral Bushen Yishou Capsule .Neurological function defects scores ,blood rheological properties and blood hs‐CRP were evaluated in all patients before and after treatment for 1 month .Results Three sets of results in treatment of limb function after scoring ,hemorheology and serum hs‐CRP levels were significantly improved (P<0 .01) .There was better improve‐ment in Combined group than these in Acupuncture group and Drug group respectively (P<0 .01) .Conclusion Yuan‐source Points and Collateral‐points combination combined with Bushen Yishou capsule can significantly improve neurological function and hemo‐rheology ,hs‐CRP levels in patients with cerebral infarction .

Objective To observe the changes of serum calcium, phosphorus, calcium-phosphorus product and para-thyroid hormone(PTH)levels after treatment with different concentrations of calcium dialysate in combination with calcitri-ol. Methods Thirty-six patients on maintenance hemodialysis were randomly divided into Dca1.25 group and Dca1.5 group, 18 cases in each group. Patients took different doses of calcitriol on the different values of PTH basis. Changes of se-rum calcium, phosphorus, calcium-phosphorus product,PTH and bone alkaline phosphatase (BAP) levels were recorded re-spectively on four time points from prior treatment and 1, 3 and 6 months of therapy. The levels of systolic blood pressure (SBP) and diastolic blood pressure (DBP) were also observed. Results There was an interaction between treatment effects and time effects in serum calcium, phosphorus, calcium-phosphorus product levels in two groups (P＜0.05). With the dura-tion of treatment, there was a trended to decrease and increase respectively in DCa 1.25 group and DCa 1.5 group, and trend-ed to stabilize for therapy 3 to 6 months. There was no interaction between serum PTH and BAP levels and time effects in two groups. There was no statistical difference in simple treatment effects and time effects. The serum PTH level showed a trend of increase gradually in DCa1.5 group with the duration of treatment. In DCa 1.25 group, the serum PTH level trended to stabilize after 3-month therapy and showed a trend of decrease gradually. There was an interaction between treatment effects and time effects in SBP in two groups, but no interaction in DBP. There was a significant difference in SBP before and after treatment in two groups. The level of SBP reduced tardily with time of treatment and reduced significantly after 3 and 6-month therapy. Conclusion The application of DCa1.25 dialysate in combination with calcitriol decreased serum calci-um effectively and kept the lower limit of normal value, which increased tolerability of active vitamin D and calcium carbon-ate therapy, and decreased SBP in maintenance hemodialysis patients.

Objective To analyze clinical features,surgical treatment efficacy and prognostic factors of bone metastasis patients after liver transplantation for hepatocellular carcinoma.Methods A retrospective clinical data of 20 bone metastasis patients after liver transplantation for hepatocellular carcinoma from July 2000 to January 2010 were received.The effect of surgery aimed at bone metastasis was evaluated.Univariate and multivariate prognostic risk factors were analyzed.Results The median survival time of these patients was 7.5 months and 1-year survival rate was only 20％.Surgical treatment could relieve pain and promote patients' peformance status significantly.Univariate and multivariate analysis found that tumor microvascular invasion within the removed recipient liver was the only prognostic risk factor.Conclusions Patients of bone metastasis after liver transplantation for HCC had poor prognosis.Surgical treatment helps improve patient's quality of life.Tumor microvascular invasion is the risk factor of surgical prognosis.

AIM: To study the effects of CIK cocultured with DC that pulsed with RCC antigen on renal carcinoma cells.METHODS: DC and CIK cells were generated respectively by cytokines from PBMC of healthy blood donor.Cell surface markers were analyzed by flow cytometry.Then CIK were cocultured with autologous DC that was(or not) pulsed with RCC antigen(786-0 cells).Cytotoxic activity against 786-0 or PC3 cells was measured by MTT assay under three different conditions: CIK cocultured with DC which was pulsed with 786-0 antigen(group A);CIK cocultured with DC which is not pulsed with 786-0 antigen(group B);CIK without DC(group C).RESULTS: The cytotoxic activity of three groups against 786-0 cells was(70.64?8.26)%,(53.40?7.33)%,(46.64?6.01)%,respectively(E/T=(20∶1)).Significant differences between group A and group B or between group A and group C were observed(P

Objective To investigate the effect of chemotherapy regimen of rituxmab combined with CHOP (R-CHOP) on the survival of patients with diffuse large B cell lymphoma (DLBCL). Methods One hundred and fifty-six cases of DLBCL diagnosed according to the WHO 2008 classification were collected from the haematopathological laboratory, the department of pathology, and Beijing University Health Science Center. Standard two-step method of immunohistochemical staining with Envision was used to assess the expression of CD10, MUM-1, bcl-6, and bcl-2. The different classification models were made according to the immuaohistochemical staining results. Hans algorithm classifies the patients into two subgroups originating from germinal center B cell-like cell (GCB) and non-germinal center B cell-like cell (non-GCB), and Muris model were classfied the DLBCL patients into the good-survival groupl and the poor-survival group2. Thirty patients with treatment of R-CHOP were set as study group and the other 126 patients without Retuxmab were defied as control group. The data were analyzed with X2 test, log-linear model and Life Table survival analysis by the SAS 8.2 statistical package. Results The 3-year survival rate of the study group was 78.3 %, but was 53.4 % in the control group. The over-all survival of the study group was obviously better than the control group with the significant difference (P <0.05). Hans algorithm showed no implication of survival for any group. The survival of different groups in Muris model has no difference in study group but was obvious in control group. The expression of bcl-2 protein has no association with survival in study group but acted as a worse implication of survival in control group. Conclusion R-CHOP chemotherapy regimen could improve the remission rate and over-all survival of DLBCL. Rituxmab could weaken the effect of bcl-2 expression in the prognosis, and the implication of survival by Muris model has diminished.

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.

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.

Objective To evaluate the efficacy of microvasoepididymostomy for the treatment of obstructive azoospermia. Methods Ninety-eight patients were confirmed as obstructive azoospermia by infertility investigations.The mean (range) age was 31 (20-43) years.The mean duration of obstruetion was 4 years.All patients were seen by the surgeon for a complete history and physical examination.Semen analyses proved azoopermia,serum levels of sexual hormone were normal and testicular function for production of sperms were normal certified by biopsy.All patients with suspected epididymal or vasal obstruction were offered scrotal exploration.The decision for microvasoepididymostomy was made during surgery,based on the pateney of the vas towards the abdomen documented by saline and sperm fragments containing in the epididymal fluid on intraoperative light-microscopic examination.Microvasoepididymostomy was performed in 58 patients documented epididymal obstruction The initial semen analysis was then done after usually 3 months postoperatively.Patency was arbitrarily defined as＞10 000 sperm/mL in ejaculate in at least one semen analysis after surgery.Postoperative patency rate and postoperative impregnation rate were followed. Results Fifty patients were followed up for 3-29 months,8 cases lost.Sperm was found by semen analysis in 36 patients.Sperm density was 4×104-2×108 sperms/ml and motility was 2%-70%.Semen analysis reveals azoospermia in 6 patients and the patients were counseled to undergo further testing to determine the ultimate outcome of the procedure.Natural conception occurred in 14 patients followed for more than 12 months.The overall pateney rate was 72%(36/50).Among patients with a follow-up of＞6 months,the natural paternity rate was 28%(14/50).The median time to achieve a natural pregnancy was 6.6 (4.0-10.0)months. Conclusion A more favourable patency can be achieved using microsurgical intussusception vasoepididymostomy and a part of patients with obstructive azoospermia can be cured by this means.