Purpose　To investigate the role of urokinase plasminogen activator (uPA),uPA receptor (uPAR),tissue type plasminogen activator (tPA) and plasminogen activator inhibitor 1 (PAI-1) in the invasiveness of human breast cancer cells.　Methods　Three human breast cancer cell lines with different invasive ability were taken as research targets.RT-PCR and milk plates methods were used to detect the expression of uPA system members and the PA activities,respectively.Modified Boyden's chamber model was employed to detect the invasive ability of cancer cell.　Results　MDA-MB-231 could express high level of uPA,uPAR,PAI-1 and low level of tPA.MDA-MB-435 could express lower level of uPA and hight level of tPA,but no PAI-1 and uPAR were detected.MCF-7 could express lower level of uPAR and high level of PAI-1,but no uPA and tPA were detected.MDA-MB-231 cells showed the highest total PA and uPA activity.MDA-MB-435 cells also showed high total PA activity,but almost all the activity owed to tPA.MCF-7 showed almost no PA activity.Correlated with their PA activities,MDA-MB-231 was found the most invasive in vitro,followed by MDA-MB-435,and MCF-7 almost had no invasive ability.The antibodies against uPA and uPAR were significantly effective in reducing the matrigel invasiveness of MDA-MB-231 by approximately 83.1% and 43.9% respectively (P<0.05).　Conclusions　Co-expression of uPA,uPAR and PAI-1 in human breast cancer highly correlates with the invasiveness in vitro.

A sandwich ELISA for the measurement of urokinase(UK)antigen was developed based on anti—UK monoclonal antibodies(McAbs)against two non—overlapping epitopes.The lower limit of sensitivity of the assay was 0.15ng/ml.Coefficients of variation of the assay at physilogi-cal levels of UK were 4.3 percent within assaysw and 8.7 percent between assays.The recovery of added UK was about 98 percent.Culture media of some human malignant cell lines contains more UK than that of normal cell lines measured by our assay.The ELISA was used to measurethe concentration of UK in plasma from 82 healthy donors.The mean value for the healthy donors was 1.31?60.6ng/ml of UK in plasma.

Objective To investigate the p53 gene mutations in Chinese patients with epithelial skin cancers. Methods Biopsies of 22 squamous cell carcinoma(SCC)and 18 basal cell carcinoma(BCC)were analyzed by polymerase chain reaction denaturing gradient gel electrophoresis(PCR-DGGE) and DNA sequencing for alterations in the exons 5～8 of p53 tumor suppressor gene. Results p53 gene mutations were found in 32.5%(13/40)of tumor tissues and 92.3%(12/13) of the mutations occurred at sun exposed areas. 36.4%(8/22)of SCC tissues and 27.8%(5/18) of BCC tissues presented p53 gene mutations. Of the 13 mutations within the exons examined, 76.9%(10/13) were missense mutations and 23.1%(3/13) were nonsense mutations. Twelve of the 13 mutations were of single nucleotide substitutions. Six of the 13 mutations involved dipyrimidine sites, where a C was replaced by a T or CC was replaced by TT(five C→T transitions and one CC→TT transition). Conclusion Ultraviolet(UV) radiation might be related to the pathogenesis of skin cancer. Mutations of p53 gene caused by solar UV might play a significant role in UV tumorigenesis, although other carcinogens might also be involved.

Objective To assess the effect of alprostadil on accelerating early recovery of renal transplant patients with delayed graft function.Methods A randomized control clinical trial was designed in renal recipients with delayed graft function(DGF)who were administered with 50μg alprostadil each day after the operation.The effects of alprostadil were compared with the control group which included 14 recipients with DGF to determine the influences of alprostadil on urine,serum creatinine(SCr).Under Doppler ultrasound,the renal blood flow resistance-indexes(RI)were measured.The rates of acute renal graft rejection(AR)were also calculated in both groups,and the 1year patient/graft survival rates were compared between the two groups.Results Urine was significantly higher in alprostadil-treated group than in control group(P ＜ 0.05).On the contrary,SCr and RI were significantly lower in alprostadil-treated group than in control group(P ＜ 0.05).No significant differences were found between the two groups in AR rate(6.25％ vs 7.14％)and the 1-year patient/graft survival rates(P ＞ 0.05).Conclusion The application of alprostadil in renal recipients with DGF had beneficial effects on accelerating recovery of renal graft function.

Objective To explore the clinical value of peripheral blood albumin gene and alpha-fetoprotein gene in early diagnosis and differential diagnosis ofhepatocellular carcinoma(HCC).Methods Two gene fragments were amplified by nested reverse transcription polymerase chain reaction(RT-PCR)with the samples from patients.Results The positive tales of peripheral blood gene fragments from patients with HCC were 53.7％in AFP and 59.7％in Alb.respectively.The former Wag signifieantly higher than that in liver cirrhosis(LC),chronic hepatitis(CH),acute hepatitis(AH)and extrahepatic tumor(ET).The latter Wag also significantly higher than that in LC,CH and ET(P

Objective To identify the antibiotic resistance, homology of imipenem-resistant Pseudomonas aeruginosa strains isolated from older in Zhejiang Hospital and the carbapenemases determinants of imipenem-resistant strains. MethodsTwo hundred and sixty-two strains of Pseudomonas aeruginosa were isolated through May 2006 to May 2009 from older in Zhejiang Hospitals. K-B method was used to determine the 16 antimicrobial agents resistance of these 262 strains. The MICs of strains to 14 antimicrobial agents were determined by agar dilution and E test method. The coding sequence of Metallo-β-lactamases (MBL) were amplified, PCR products were purified, cloned and sequenced. The homology of these isolates was analyzed by pulse-field gel electrophoresis(PFGE). ResultsOne hundred and four strains of imipenem-resistant Pseudomonas aeruginosa were screened from 262 strains. The resistant rates of 104 isolates of Pseudomonas aeruginosa to ampicillin/sulbactam and cefoperazone/sulbactam were 78.9％ and 35.9％ ; polymyxin E had a minimal resistance of 6.0％ ; minocycline had a resistance rate of 58.3％. The resistant rates to other antimicrobial agents were more than 70.0％. Twelve imipenem-resistant Pseudomonas aeruginosa strains contained MBL gene and two kinds of integron were detected from 10 of these 12 strains. Twelve strains of Pseudomonas aeruginosa belonged to 5 epidemic PFGE-clone. ConclusionAll of the imipenem-resistant Pseudomonas aeruginosa which had cause outbreaks in Zhejiang Hospital. MBL were not the most popular gene type. All of the MBL gene types were VIM-2. The blaVIM-2 gene cassettes located in diflerent class 1 integrons. The integrons dissemination was the most important style of strains spread.

Objective To investigate the security and result of operation of one side polycystic kidney removal and homonymy kidney transplantation simultaneously for giga-polycystic kidney di-sease of terminal stage. Methods Forty-five patients with polycystic kidney of transplantation were retrospectively analyzed. The patients were divided into 2 groups. Patients of group A (n=23) under-went resection of the cystic kidney by extraperitoneum and the other 22 patients(group B) didn't re-move the cystic kidney. The data including average length of hospital stay, variance of blood pressure, lessen of abdominal circumference, lung capacity, total lung capacity, FEV1.0/FVC, incidence rate of delayed graft function (DGF) and 1 year patient/kidney survival rate of the 2 groups were compared. Results The average length of post-operative hospital stay of group A was (14.5±2.6)d,lessen of blood pressure was (30.0±0.7/13.34±8.4)mm Hg, lessen of abdominal circumference was (11.0+ 6. 3) cm, lung capacity increased (1.4±0.3)L, total lung capacity increased (2.0±1.0)L, FEV1.0/ FVC increased (5.3±1.0) %, the incidence rate of DGF was 8.7% (2/23), 1 year patient/ kidney survival rate was 100.0%/95.7%. The average length of post-operative hospital stay of group B was (28.45±7.9)d,lessen of blood pressure was (3.9±11.2/2.9±12. 0)ram Hg, lessen of abdominal circumference was (3.3±2.2)cm, lung capacity increased (0.44±0.3)L, total lung capacity increased (0.8±0.2) L, FEV1.0/FVC increased (2.0±0.9)%, the incidence rate of DGF was 9.1%(2/22), 1 year patient/kidney survival rate was 100.0%/95.5%. There were no significant differences of the incidence rate of DGF and 1 year patient/kidney survival rate between the 2 groups. While there were significant differences of the other data between the 2 groups(P＜0.05). Conclusions It is safe and convenient for one side polycystic kidney removal and homonyrny kidney transplantation simultaneous-ly for giga-polycystic kidney disease of terminal stage. The procedure could be applied to the patients of graveness complication or giga-polycystie kidney hampering operation of transplantation.

OBJECTIVE: Complication incidence of urinary fistula which frequently occurs following renal transplantation is 3%-10%. Thus, poor processing may cause loss of transplanted kidney. This study was designed to retrospectively analyze urinary fistula following renal transplantation and to summarize the processing experience. METHODS: A total of 27 out of 1 203 patients with urinary fistula following renal transplantation (16 males and female 11 and mean age of 43 years) were collected from Guangzhou General Hospital of Guangzhou Military Command of Chinese PLA from December 2000 to March 2009. Totally, 26 patients accepted donor kidney from corpse, and 1 from living body. All patients were treated with expectant treatment (n=17) including inserting 18Fr Foley catheter alone and draining from the native drainage channel of operative site (n=12) and inserting a ureteral stent (single-J) by cystoscope retrograde approach and inserting 18Fr Foley catheter into the bladder (n=5); exploring operation (n=10) including anastomosing ureter and bladder and placing ureteral stent (n=5) and anastomosing ureter and ureter of recipient and placing ureteral stent (n=5); pedicled omentum grafts to cover and surround stoma after suturing (n=6). RESULTS: Only 1 case was failed because kidney vain was injured in the second operation and the kidney was resected. Another 26 cases were cured. Within the 3 month to 7 years follow-up, the urinary fistulas did not relapse, no stegnosis or hydronephrosis, no urinary tract infection and renal function were normal. CONCLUSION: Rapid diagnosis and treatment for urinary fistulas after renal transplantation is imperative. First mostly patients may be cured by expectant treatment. If not then perform exploratory operation. Using peclicled omentum grafts to cover and surround stoma after suturing for complex urinary fistulas can raise achievement ratio of operation.

Objective To investigate the genotypes of β-lactamases produced by Klebsiella pneumoniae.Methods Plasmid conjugation,PCR amplification,gene cloning and DNA sequencing,isoelectric focusing electrophoresis and extended-spectrum β-lactamase(ESBLs)confirmatory test were carried out for analyzing the encoding gene of β-lactamases in clinical strains of Klebsiella pneumoniae collected from hospital wards.Results Totally 75 clinical strains of Klebsiella pneumoniae were collected,in which 48 strains were confirmed to produce genotype of β-laetamases(64.0%),including 39 ESBLs-producing Btraim(52.0%).Among 48 strains,17 isolates(35.4%)carried 2 types of ESBLs genes,7(14.6%)carried 3 types of ESBL8 genes,and 5(10.4%)carried 4 types of ESBLs genes.CTX-M was the most comon type(30/48,62.5%),followed by TEM(26/48,54.2%)and SHV(25/48,52.1%).Among 9 isolates with DHA-1 AmpC β-laetamase,8 produced AmpC β-lactamases and ESBLs.Class A carbapenemase KPC-2 was produced in 3 isolates.False negative rate of ESBLs confirmatory test was 23.1%(9/39).Condusion Genotypes of β-lactamases produced by Klebsiella pneumoniae are complicated,which results in multi-drug resistance in clinic.

Objective To investigate the coverage of a recombinant protein vaccine based on pneumococcal surface protein A (PspA) from both family 1 and family 2.Methods One hundred and fifty-nine Streptococcus pneumoniae strains, including 47 invasive strains, were isolated from children in Nanjing Children′s Hospital.Cell lysates were prepared and reacted with three antibodies recognizing PspA -RX1, PspA-3296 and PspA-5668 for PspA typing by ELISA .Results Among 47 invasive isolates of 9 different serotypes, 10.7%were PspA family 1 and 89.3%were PspA family 2.Among all of 159 clinical isolates, 10.1% were identified as PspA family 1, 88.0%were family 2, while 1.9%of strains could not be typed by ELISA and PCR assays .None of strains belonged to PspA family 3.Conclusion The recombinant pro-tein vaccine based on PspA from both family 1 and family 2 has a broad coverage among clinical isolates and is potentially protective against both invasive and non-invasive pneumococcal diseases .