BACKGROUND:Cancer stem cell (CSC) hypothesis suggests that tumorous clones are maintained by a rare fraction of cells with stem cell proprieties. Several kinds of CSCs of solid tumor have been isolated in recent years. However, there have been fewer studies on the objective existence of bladder cancer stem cells (BCSCs) and on the methods to effectively isolate and identify BCSCs. OBJECTIVE:To investigate possibilities of BCSC existence and of epithelial membrane antigen (EMA) used as a surface marker of BCSC. DESIGN:A control observation experiment. SETTING:Tianjin Institute of Urinary Surgery & Second Hospital of Tianjin Medical University. MATERIALS:This study was performed at the Room for Tumor Immunity of Tianjin Institute of Urinary Surgery (key laboratory for State "211 Project") from March 2006 to July 2007. Nine specimens of human bladder were obtained from patients who received treatment in the Second Hospital of Tianjin Medical University. These specimens corresponded to the diagnostic criteria of low malignant potential papillary urothelial neoplasm and low-grade papillary urothelial carcinoma. Additionally, 40 samples of human low malignant bladder transitional cell carcinomas (BTCC) and 10 samples of normal urothelium that were used for immunohistochemistry were obtained from the patients who received treatment in the Department of Urinary Surgery, Second Hospital of Tianjin Medical University. Written informed consent for the specimen providing was obtained from the patients, and the protocol was approved by the hospital’s Ethics Committee. METHODS:The genes that were differentially expressed between normal urothelium and BTCC were identified through a DNA array assay to preliminarily determine the existence of BTCC. Overpressed stem cell related genes, Bmi-1 and EZH2, were verified by immunohistochemistry. A total of 27 potential surface markers of BCSCs were assayed to determine the location of positive cells. EMA- subsets were obtained through an immunomagnetic bead cell sorting system to analyze their abilities for colony forming, self-renewal and extensive proliferation. MAIN OUTCOME MEASURES:Normal urothelium and BTCC gene expression difference, Bmi-1 and EZH2 protein expression difference between low malignant BTCC and normal urothelium; and experimental results of colony forming. RESULTS:A total of 268 genes (including Bmi-1 and EZH2) that were differentially expressed between normal urothelium and low malignant BTCC were identified through a DNA array assay. The Bmi-1 and EZH2 had been found overexpressed in the low malignant BTCC. Except for EMA, above-mentioned 26 out of 27 potential surface markers were null for isolating BCSCs. EMA- subsets were located in the basal layer and suprabasal epithelial layers of both normal and tumorous urothelium (potential location of BCSCs). EMA- subsets possesed the abilities for colony forming, self-renewal and extensive proliferation. CONCLUSION:The experiment confirms the existence of BCSCs. EMA- might be a surface marker of BCSCs.

ObjectiveBy analyzing the transfusion-related data using DRGs data to establish a credible evaluation system for clinical use,thereby enhancing the safe and effective clinical transfusion management.Methods Use the transfusion-related data of DRG-s,and statistical methods of EXCEL database to categorize and analyze the number of blood transfusion,hospital name,hospital type,hospital level,DRGs code and name,times of blood transfusion,volume of transfusion,etc.Results Output the statistics of the city's ratio of institutional use of blood,per capita consumption in patients,distribution and ordering of hospital based blood tranfusion,distribution and ordering of DRGs based blood transfusion,distribution and ordering of single DRGs between hospitals,annual consumption growth in different hospitals and major disease groups of blood tranfusion.ConclusionThe use of DRGs can efficiently control clinical indicators for blood transfusion and evaluate the performance of blood transfusion.it can provide a credible management tool for the health administrative departments and hospitals while offering data support for policy making of management objectives at the same time.

Antimutagenic effect of p-aminobenzoic acid (PABA) and sodium bisulfite (NaHSO_3)on the mutagenicity of N-methyl-N-nitro-N-nitrosoguanidine (MNNG) was detected by the mehod of human lymphocytes unscheduled DNA Sythesis (LIDS). The results showed that when the concentration of PABA and NaHSO_3 was varied in presence of a constant concentration of MNNG, 5 ?10~(-5) mol/L concentration of PABA, 3 ? 10~(-4) mol/L concentration of NaHSO_3 were the most effective, while the concentration of MNNG varied in the presence of a constant concentration of PABA and NaHSO_3, antimutagenicity was the most effective at high concentration of MNNG. The results indicated that the PABA and NaHSO_3 exhibited antimutagenic activity towards MNNG-induced mutagenicity in LIDS and the extents of antimutagenicity were related with the concentration of MNNG.

Objective To study the surgical treatment of multilevel arterial occlusion of lower extremities. Methods Single or jumping arterial bypass using grafting and percuteneous transluminal angioplasty(PTA)and stenting combined with infrainguinal revascularization were used for the multilevel arterial occlusion in 25 lower extremities. Results 25 cases showed satisfactory results in 6 months to 3 years follow-up. Postoperatively,the ischemia symptoms of limbs were improved or disappeared,4 toes were amputared because of preoperative dry gangrene,the wounds received eurement in a 2 to 4 months,2 graftings became occluded in 6 months postoperation. There were no procedural or postoperative morbidity or mortality.The cumulative patency rate of grafting was 92%(23/25),The rate of curement was 100%.Conclusion According to the individual principle,it is an effective therapy method to choose single or jumping arterial bypass and PTA and stenting combined with infrainguinal revascularization for high risk patients with multilevel arterial occlusion.

Objective To appraise the postoperative anti-reflux function of vertical esophagogastic valve-plasty anastomosis for cardiac cancer.Methods Forty patients with cardiac cancer were randomly divided into study group and surgery control group,with 20 patients in each group.The study group underwent vertical esopagogastric valve-plasty anastomosis,while the surgery control group underwent conventional esophagogastrostomy.Ten healthy volunteers were recruited as normal control group.A 24-hour esophageal pH monitoring and endoscopy check-up was carried out in all experimental subjects at 90 days postoperatively.Results All of the pH monitoring indexes in study and surgery control groups were higher than those in normal control group(P

Objective To investigate the clinical and pathological characteristics of nephrogenic adenoma. Methods Eleven patients were diagnosed as nephrogenic adenoma including 5 men and 6 women, aged 37-78 years (56 on average). The pathological findings in all cases of nephrogenic adenoma were presented with a review of the literature. Results Eleven cases of nephrogenic adenomas were evaluated, 2 cases were in ureter and 9 cases were in the bladder. Eight of the 9 bladder cases underwent TUR-BT surgery in continuous epidural anesthesia, 1 case underwent partial cystectomy with general anesthesia. A right ureteroscopy and left ureterolithotomy were performed respectively in continuous epidural anesthesia for the 2 cases in ureter. The final diagnosis was based on histopathological findings. For all of cases, 8 cases were diagnosed as nephrogenic adenomas, 2 cases as atypical nephrogenic adenoma and 1 case as nephrogenic adenoma with malignant transformation. The microscopic appearance of nephrogenic adenoma demonstrated that morphology closely resembled aberrant tubules of the kidney. In addition, atypical nephrogenic adenomas appeared as the presence of cytologic atypia, including nuclear enlargement, nuclear hyperchromasia and prominent nucleoli. The morphologic changes of nephrogenic adenomas with malignant transformation were that tumor cells retained the basic structural characteristics of typical nephrogenic adenomas, and the similar morphological cells lost adhesion ability among cells and presented diffuse solid growth in the surrounding area.Intravesical perfusion was further performed for treating the patients with atypical nephrogenic adenomas or nephrogenic adenomas with malignant transformation. The mean patient follow up was 46 months (range, 24- 104 months), and there was only 1 case of recurrence. Conclusions Nephrogenic adenoma is an uncommon benign lesion of the urinary tract. The symptoms and cystoscopic manifestations are not unique. We reported one patient of nephrogenic adenomas with malignant transformation and provided some evidence for malignant alteration in morphology and invasive behavior. All patients underwent local excision of the lesions. Intravesical perfusion was further performed for treating the patients of atypical nephrogenic adenomas or nephrogenic adenomas with malignant transformation. Whether it is nephrogenic adenoma or atypical nephrogenic adenoma, long-term follow-up after treatment is necessary.

Objective: To construct the lentiviral plenti6/V5-D-TOPO vector containing TβR Ⅱ Dnglytk and control vector by developing a lentivirus mediated gene transfer program incorporating a herpes simplex virus thymidine kinase(HSVtk) in the dominant negative TGF-β type Ⅱ receptor (TβR Ⅱ Dnglytk) expression vector. Methods: The PCR methods were used to amplify the genes TβR Ⅱ DN and HSV-tk from the respective plasmids. Then the genes were Linked by recombinant PCR technology to construct the fusion gene TβR Ⅱ Dnglytk and control vector TRANSglytk. According to the operation manual (from the Invitrogen company), ToPe cloning technology were used to construct the plasmids of plenti6/VS-D-TOPO(R)-TβRⅡ Dnglytk and plenti6/VS-D-TOPO -TRANSglytk. Both of the constructed plasmids were verified by sequencing.Results: The constructions of the plasmids of plenti6/V5-D-TOPO(R)-TβR Ⅱ Dnglytk and plenti6/V5-D-TOPO(R)-TRANSglytk were completed smoothly. The DNA sequencing results showed that both the plasmids were constructed correcdy and can be used in the production of infectious lentivirus vectors. Conclusion: Using TOPO cloning technology in the construction of the plasmids of plenti6/VS-D-TOPO -TβR Ⅱ Dnglytk and plenti6/VS-D-TOPO(R)-TRANSglytk and recombinant PCR in the fusion genes are feasible. The recombinant PCR combined with ToPe cloning technology can be the simple, highly efficient and rapid way to construct lentiviral vector and construction of plenti6/V5-D-TOPO(R)-TβR Ⅱ Dnglytk, which will lay a foundation for tumor immunotherapy.

Objective To investigate the myocardial protective effects of trimetazidine by observing the changes of peripheral B-type natriuretic peptide (BNP),cardiac troponin I (cTnI) level in patients undergoing off-pump coronary artery bypass graft (OPCAB), and the clinical significance of peripheral cTnI and BNP in cardiac surgery. Methods One hundred and three OPCAB patients were divided into trimetazidine group (52 cases) and control group (51 cases) by random digits table. The serum levels of BNP and cTnI preoperatively,postoperatively of 24 h, 72 h and 7 d were detected. Results The serum levels of BNP [(224.5 ± 12.0), (331.2 ±22.6), (82.4 ±3.3) ng/L] and cTnI [(0.21 ±0.04), (1.32 ±0.49), (0.26 ±0.04) μ g/L] in trimetazidine group were lower than those in control group [(294.7 ± 11.8 ), ( 383.9 ± 28.3 ),( 112.4 ± 12.5 ) ng/L and ( 1.20 ± 0. 13 ), (2.35 ± 0.54), (0.75 :± 0.21 ) μ g/L] postoperatively of 24 h, 72 h and 7 d (P＜ 0.05 ). The serum levels of BNP and cTnI increasedd at 24 h after operation. The peak level was found at 72 h and remained higher level until 7 d after operation. The baseline levels of BNP were positively correlated with cTnI (r = 0.635,P ＜ 0.05), but negatively correlated with left ventricular ejection fraction (LVEF) (r =-0.674,P ＜ 0.01 ). Conclusion Trimetazidine can obviously reduce serum levels of BNP and cTnI in patients undergoing OPCAB. So the united application of serum BNP and cTnI may be as a monitor marker to reflect the cardiac function in patients after OPCAB.

Objective To evaluate the safety and clinical effects of radiofrequency single catheter ablation (RESCA)for right ventrieular arrhythmia(RVA).Method A total of 111 patients data in the Second Affiliated Hospital of Wenzhou Medical College from May 2003 to May 2008,were retrospectively analyzed aged(45.2±16.7)years old including 41 men and 70 women,consisted of 13 patients of ventricular tachycardia(VT)and 98 patients of premature ventricular contractions(PVC).There were 104 casess from right ventricular outflow tract arrhythmia(RVOTA)and 7 cases from right ventricular inflow tract arrhythmia(RVITA).According to use single catheter approach or common technique,electrophysiolo-gical study,pacing and/or activation mapping and Catheter ablation were performed,were separated into two groups.①Single catheter group:27 men and 49 women,ages(44.5±16.9)years old;consisted of 62 patients of RVOT-PVC,9 patients of RVOT-VT and 5 patients of RVIT-PVC.②Control group:14 men and 21 women,ages(46.7±16.5)years old;consisted of 29 patients of RVOR-PVC,4 patients of RVOT-VT and 2 patients of RVIT-PVC.Results Operations in two groups came off smoothly and no ablation related complications in two groups.Procedure time and fluoroscopy time[(55.23±26.24)min and(9.93±5.32)min]in single catheter group were significantly shorter than those in control group [(68.37±21.83)min and(12.96±4.54)min,t=2.76 and 3.09,all P<0.01].Cost in the fromer (12440.32±761.24)RMB were significantly less than those in the latter[(22119.51±1071.07)RMB,t=46.09,P<0.01].Ablated successful rate in the near future,at a specified future date and other parameter were similar in two groups.Conclusions Right ventricular arrhythmia can be ablated with single catheter approach in safety,efficacious,easy to operate and lower cost.

Objective Our hypothesis was if we rendered host' s cytotoxic tumor lymphocytes insensitive to TGF-β,these immune cells could be able to overcome the TGF-β mediated immunosuppression and reject the tumor.We aimed to develop a lentivirus mediated gene transfer program incorporating a herpes simplex virus thymidine kinase(HSV-tk)in our dominant negative TGF-β type Ⅱ receptor(TβRⅡDNglytk)expression vector.So we first need to construct the lentiviral pLenti6/V5-D-TOPO vector containing TβRⅡDNglytk and produce the recombinant lentivirus as the transfection vector.Methods PCR were used to amplify the genes TβRⅡDN and HSV-tk from the respective plasmids.Then the genes were linked by recombinant PCR technology to construct the fusion gene TβRⅡDNglytk and control vector TRANSglytk.According to the operation manual from the Invitrogen company,TOPO cloning technology was used to construct the plasmids of pLenti6/V5-D-TOPO(R)-TβRⅡDNglytk and pLenti6/V5-D-TOPO(R)-TRANSglytk.Both of the constructed plasmids were verified by sequencing.ViraPowerTM Lentiviral System and 293 FT cells provided by Invitrogen were used to produce the recombinant lentivirus vector,the tillers of the lentivirus were determined by 293 cells.Results The construction of the plasmids of pLenti6/V5-D-TOPO(R)-TβRⅡDNglytk and pLenti6/V5-D-TOPO(R)-TRANSglytk were completed succefully.The DNA sequencing results showed that both the plasmids were constructed correctly.Using them we successfully produced infectious lentivirus vectors with appropriate tilters.Conclusions Using TOPO cloning technology in the construction of the plasmids of pLenti6/V5-D-TOPO(R)-TβRⅡDNglytk and pLenti6/V5-D-TOPO(R)-TRANSglytk and recombinant PCR in the fusion genes is feasible.Recombinant PCR combine with TOPO cloning technology can be a simple,highly efficient and rapid way to construct lentiviral vector.The production of infectious lentivirus with appropriate tilters using 293FT is suitable and feasible.The construction of pLenti6/V5-D-TOPO(R)-TβRⅡDNglytk and production of the infectious lentivirus will lay a foundation for immunotherapy of prostate cancer.