[Objective]To determine the biological effects of transfection of adeno-associated rirus(AAV) BMP4/7 on rabbit bone marrow stromal cells (BMSCs).[Method]The mature peptide of BMP4 and BMP7 were gained by One-Step RT-PCR from the human placenta and the BMP4/7 fusion gene was gained through gene recombinant techniques and then transferred to pGEM plasmid. The BMP4/7 fusion genen was cut down from the pGEM-BMP4/7 and the recombination was successfully completed in colibacillus, and recombinant adeno-assosiated was produced in 293 cells. Rabbit BMSCs were transfected with the recombinant adeno-assosiated virus vectors carrying BMP4/7 fusion gene with differen multiplicitv of infection(MOI) values. Cell growth curves were drawn to evaluate the biologic effect of AAV-BMP4/7 on cyto-activity. The transfection efficiency was measured using MTT method. The ossification of cells was evaluated by investigating the shape change of the cell ability of ALP and OC at 7,14 days after transfection. Cells were then transfected with AAV-BMP4/7 and AAV-EGFP,respectively.[Result]1.We successfully constructed the recombinant adeno-assosiated virus with BMP4/7 fusion gene.2.The ttransfection efficiency of AAV-BMP4/7 was roughly 72% without siginifficant biologic effect on cyto-activity.The ossification of cells was significant after transfection with AAV-BMP4/7. The 1?105 vg/cell MOI value of transfection efficiency was stroger than 5~104 vg/cell MOI value (59.3,8%). 3. The concentrations of ALP and OC in AAV-BMP4/7 transfection groups were significantbly higher than in AAV-EGFP groups (tALP=896.88 P

Objective To observe the expression change of HRG-1 gene between urinary bladder carcinoma and normal tissues,and to investigate the effect of HRG-1-siRNA on the proliferation and apoptosis of human bladder carcinoma cells.Methods Immunohistochemisty was used to detect the expression of HRG-1 in 85 cases of bladder carcinomas and 20 normal bladder tissues.The siRNA of HRG-1 was designed,synthesized,and transfected into bladder cancer cell line T24.Results The HRG-1 gene expression had significant differences between bladder carcinoma and normal bladder tissues (P ＜ 0.05).The positive expression of HRG-1 gene had significant differences between the pathological grades and clinical stages of bladder carcinomas (P ＜0.05).After treated with siRNA,the expression levels of HRG-1 protein and mRNA in T24 cells decreased obviously (P ＜ 0.05).The apoptosis rate of T24 cells transfected with HRG-1-siRNA was significantly different from control-siRNA group and blank group (P ＜ 0.01).Conclusions The high expression of HRG-1 gene may play an important role in bladder carcinoma,and siRNA targeting HRG-1 can suppress HRG-1 protein expression markedly and enhance apoptosis of T24 cells.

Objective To explore the expression of a new candidate tumor suppressor N-myc downstream regulated gene 2 (NDRG2) in bladder cancer tissues and to investigate its clinical and pathological significance.Methods Formalin-fixed,paraffin-embedded tissue sections from 62 cases of bladder carcinomas and 10 cases of normal bladder tissues were analyzed retrospectively with immunohistochemistry (S-P method).Results The NDRG2 gene was highly expressed in normal bladder tissues,but low expressed in bladder carcinoma tissues.Positive expression of NDRG2 was detected in 8 of the 10 (80.0％) normal tissues and 40.3％ in bladder carcinoma ones (x2 =3.98,P ＜0.05).Furthermore,with the degree of malignancy increased,the positive expression of NDRG2 in bladder carcinoma samples was decreased.The expression of NDRG2 in bladder carcinoma was negatively correlated(r =-0.288,P ＜0.05) with C-myc(r =-0.436,P ＜0.01) and positively correlated with p53 in bladder carcinoma tissues(r =0.717,P ＜0.01).Conclusions The level of NDRG2 expression was lower in bladder carcinomas than in normal tissues.NDRG2 may play an important role in bladder carcinogenesis and in the progress of bladder cancers.

Objective To study the hemostnsis effects of compression cords oppressing after femoral artery puncture and encheiresis. Methods Self-made compression cords were used to stop bleeding of femoral artery puncture sites in 2164 patients underwent 3046 cases of arteriopuncture encheiresis. The total eases were divided into group A (1482 eases) and group B (1564 cases) according to different treatment of oppression and breaking duration which were 8-10 hours for group A and 5-6 hours for group B, the hemostasis effects, discomfortableness and related complications within two groups were evaluated. Results The proportion of patients who got good hemostasis effects were 99.2% in group A, 99.0% in group B. There was no difference in hemostasis effects between two groups (P> 0.05). While the rate of discomfortableness in group A (63.8%) was higher than that in group B(42.9%) (P<0.05). The related serious complications were 2 patients in group A, but no occurred in group B. Conclusion Self-made compression cords op- pressing and 5-6 hours breaking time course can achieve both finer hemostasis effect and lower frequency of complications after femoral artery puncture and encheiresis.

Objective To evaluate the serum pyridinoline cross-linked carboxyteminal telopeptide of type Ⅰ collagen ( ICTP) in the early diagnosis potency,efficacy and prognosis of tumor bone metastasis. Methods According to emission computed tomography(ECT) ,MRI and X-ray results,336 cases of tumor were divided into higher ICTP (5. 98 ± 1. 95μg/L ) than normal values. Twenty-two cases were identified bone metastasis through PET/CT examination. 26 cases were identified bone the effective cases decreased from( 13. 22 ± 4.65)μg/L (before treatment) to (7. 18 ±3. 54)μg/L (after treatment) (t = 10. 076,P = 0. 000). Conclusions Serum ICTP is helpful for the early diagnosis, screening and efficacy evaluation of tumor bone metastasis. It could be used for monitoring the occurrence of tumor bone metastasis and its prognosis.

Objective To investigate the antitumor activities of adenovirus-mediated NDRG2 gene (Ad-NDRG2) and docetaxel on human prostate cancer DU145 cells.Methods The protein expressions of cyclin D1,cycliu E,and NDRG2 in the cells were determined by Western blot.MTT and flow cytometry were used to observe the effects of docetaxel (10-6 mol/L,10-7 mol/L,and 10-s mol/L) and Ad-NDRG2 on prostate cancer cell line DU145 in single or synergistic administration ways for 24 and 48 hours in vitro.Male BALB/C-nu mice with DU145 prostate cancer cell lines were treated by docetaxel and Ad-NDRG2 singly or synergistically in vivo.Results After infected by adenovirus,the protein expression of NDRG2 increased,but cyclin D1 and cyclin E decreased in DU145 cells.Ad-NDRG2 inhibited the cell growth (inhibition ratio =41.8％,t =4.18,P ＜0.01),promoted apoptosis (apoptosis ratio =32.4％,x2 =11.66,P ＜0.05),changed the ratio of G2/M phase from 50.2％ to 23.6％,and reversed partially the G2/M arrest,of DU145 cells induced by 10-7 mol/L docetaxel.In vivo experiment showed that docetaxel,Ad-NDRG2,and combination of docetaxel and Ad-NDRG2 inhibited tumor growth with a inhibition rate of 30.7％,28.2％,and 55.8％,respectively.The coefficient of drug interaction (CDI) of docetaxel and Ad-NDRG2 was 0.89.Conclusions Ad-NDRG2 can enhance the growth suppression and apoptosis induced by docetaxel in synergistic way in vitro and in vivo.It demonstrated the great potential of Ad-NDRG2 in the treatment of androgen-independent prostate carcinoma.

Objective To study the clinical efficacy of laparoscopic retroperitoneal lymph node dissection (LRPLND) for stage Ⅱ nonseminomatous testicular cancer. Methods Seven patients (mean ages 28 years, 4 had the tumors on the left side and 3 on the right) underwent LRPLND during 1 to 4 weeks after orchiectomy for clinical stage Ⅱ nonseminomatous testicular cancer. The dissection-al boundaries included the ipsilateral ureter laterally, the renal vessels superiorly, abdominal aorta and the bifurcation of the iliac artery inferiorly. The operative time, intraoperative blood loss, intra-or postoperative complications, tumor markers and ejaculation were observed. Results LRPLND was completed in all patients. Operative time ranged from 140 to 220 min (mean 180 min). Intraoperative blood loss ranged from 80 to 127 ml (mean 95 ml) and no transfusions were required. No intra- or postoperative complications occurred because of the procedure. At 2 weeks after operation, serum al-pha fetoprotein decreased from (15 to 1247 μg/L) to (2 to 8 μg/L), and beta human chorionic gona-dotropin decreased from (5 to 59 μg/L) to (0.5 to 2.5 μg/L). All patients recovered ejaculation dur-ing 10 to 12 weeks after operation. After a mean follow-up period of 21 months (range 12 to 26), no disease recurrence or metastasis was observed. Conclusion LRPLND is a safe, effective, minimally invasive procedure in the management of stage Ⅱ nonseminomatous testicular cancer patients.

Due to the ability of overcoming both the dimensionality and the collinear problems of the spectral data, partial least squares ( PLS ) is in ever increasingly used for quantitative spectrometric analysis, especially for near-infrared spectrum, mid-infrared spectrum and Raman spectrum. In this work, an improved PLS algorithm is proposed for efficient information extraction and noise reduction. The spectral variables are clustering to several subsets, and several sub-models are built for each subset. Then, the sub-models are re-weighted and ensemble to the final model. Experiments on two near-infrared datasets ( octane number prediction in gasoline and nicotine prediction in tobacco leafs ) demonstrate that the new method provides superior prediction performance and outperformed the conventional PLS algorithm, and the root mean square error of prediction ( RMSEP) is reduced by 32% and 22%, respectively.

BACKGROUND: The nutritional support, as well as the water and electrolyte balance during the perioperative period in the renal transplantation recipients at diuresis stage are important to the functional restoration of transplanted kidneys.OBJECTIVE: To explore the method and opportunity of the nutritional support and the handling of the water and electrolyte balance in perioperative period of renal transplantation.DESIGN, TIME AND SETTING: A retrospective clinical analysis was performed in the Department of Urology, Xijing Hospital from June 2003 to June 2007.PARTICIPANTS: Ninety-six patients of chronic renal failure underwent allograft renal transplantation. They comprised 59 males and 37 females, aged 17-67 years, with a mean of 35.7 years.METHODS: The perioperative physiological features of the renal transplantation recipients were summarized retrospectively. The recipients' condition during the perioperative period was divided into two stages at the opening point of allograft blood current. The vital signs of the patients maintained at a stable level before operation. All patients received blood transfusion since the operation began, and were supplemented with albumin before opening the vessels. Urinary production exceeding 100 mL per hour indicated the beginning of fluid replacement, which was a simplified transfusion for the patients at diuresis stage following renal transplantation.MAIN OUTCOME MEASURES: Blood inosine, urea nitrogen, electrolyte, blood sugar and urine of the patients were detected at one day postoperatively.RESULTS: During 12-16 hours postoperatively, the urinary production was 260-1 200 mL, average 520 mL per hour. Blood routine test showed 8 cases developed mild hyponatremia, accounting for 8.3%, 3 cases occurred high potassium and healed after renal functional recovery, 1 case presented low potassium and healed with supplement therapy. There were no abnormal changes of blood chlorine. The blood glucose among 21 cases (21.9%) was higher than the normal level, and recovered following hormone maneuver. The electrolytes and blood glucose were detected to be normal in other patients, without any case with low calcium or magnesium. The urine specific gravity arranged during 1.010-1.015.CONCLUSION: The colloid such as erythrocytes, blood plasma and albumin should be mainly infused before the opening of allograft blood current. And the water and electrolytes is recommended to administrate promptly and regularly during the diuresis stage. The healing of the stoma benefits from the adequate nutritional support. The metabolic acidosis still should be prevented when the urinary production returns normal.

Aim To construct the fusion gene of human IgG3 upper hinge region and p53 tetramerization domain and to analyze its space conformation. Methods The fusion gene was obtained by recursive polymerase chain reaction (R-PCR),and was cloned into vector pUC19. The positive clone was selected and sequenced with PE310 auto-sequencer. The space conformation of the fusion gene expression product was predicted by using computer program Antheprot. Results Restriction endonuclease digestion confirmed that the fusion gene has been inserted correctly into the vector. The result of sequencing showed that the fusion gene is identical with designation. Analyzing with antheprot program showed that the fusion gene expression product could auto-assembly into a tetramer with four long and flexible linkers. Conclusion Successful construction of the fusion gene mentioned above laid the foundation for further preparation of multivalent gene engineering antibody.