ObjectiveTo established a cell line that expresses hBD1 stably,and detected the antimicrobial activity of the hBD1 to the muhidrug resistant bacterial strains.MethodsRecombinant plasmid was introduced into COS-7 cells by lipofectamine,cells were selected in culture medium containing G418 to acquired the monoclonal cell lines,total RNA were extracted from the cultured cells,expression levels of hBD1 mRNA was identified by RT-PCR,collected the supernatant solution of the cultured cell,expression levels of protein was identified by Western blot.Put the expression products and resistant organisms mixed together,after incubation in different times in 37℃,coating the mixtures in LB flat,then obtained the ratios between colonies number of experimental groups and colonies number of control groups,put those ratios as the survival rate of the drug resistance bacterias.Results The monoclonal cell lines had obtained after screened with G418,the hBD1 gene could be detected both at transcriptional and protein levels,Under the influence of expression product hBD1,survival rate of muhidrug-resistant Acinetobacter baumannii,multidrug-resistant Escherichia coli and multidrug-resistant Klebsiella pneumoniae could reduced to 9%,22% and 50%,but survival rate of multidrug-resistant Stenotrophomonas maltophilia is not have apparente difference with the control group.ConclusionThe stably-transfected cell line of hBD1 was successfully constructed,and the expression products of hBD1 showed the antimicrobial activity toward multidrug resistant bacterial strains.

OBJECTIVE To find the active principle in Radix Scutellariae,Radix Sophorae Flavescentis and Flos Lonicerae in Gansu inhibiting yeast-like fungi of deep infection.METHODS Radix Scutellariae,Radix Sophorae Flavescentis and Flos Lonicerae were separately cleaned,toasted and grinded to fine power.Five kinds of(8 strains) yeast-like fungi that were collected and spread into 90mm Sabouraud plate.On aseptic condition,bored holes on Sabouraud plate with the drilling instrument of 6mm diameter,and filled every holes with 25.73mg fine powder of Radix Scutellariae,Radix Sophorae Flavescentis and Flos Lonicerae,and added 3 to 5 drops of distilled water in the medicinal powder.Under 35℃,the bacteria were cultured for 24 to 48 hours,and the size of bacteriostatic ring was observed.RESULTS The diameters of the bacteriostatic ring of Radix Scutellariae powder inhibiting five kinds of yeast-like fungi were separately 16.0mm,13.5mm,13.0mm,13.0mm and 10.5mm,respectively,and the diameter of Radix Sophorae Flavescentis and Flos Lonicerae powder was 0.CONCLUSIONS There is the active principle inhibiting yeast-like fungi in Radix Scutellariae,but not that in Radix Sophorae Flavescentis and Flos Lonicerae.

Objective To discuss the application value of CT angiography (CTA) in evaluating renal artery anatomy of laparoscopic nephron-sparing surgery (LNSS) before operation. Methods Retroperitoneal LNSS was performed in 87 patients with renal tumors. Forty cases underwent CTA before LNSS, volume rendering and maximum intensity projection of renal artery were used as three-dimensional reconstruction. The other 47 without CTA were compared as control. Treatment out-comes of the 2 groups were compared, including operation time, warm ischemia time, blood loss, con-version to open surgery, hospitalization stay after operation, and complications after operation. Re-sults All the procedures were completed laparoscopically with no conversion to open surgery in CTA group. One patient underwent conversion to open surgery in the control group. In CTA and control group, the operation time was 94 min(range, 76-118)and 115 min(90-190, P<0.05). The warm ischemia time was 23 min(12-39)and 29 min(18-40, P<0.05). Blood loss was 90 ml(9.0-160) and 130 m1(90-600, P<0.01). Hospitalization stay after operation was 4.1 d(3-5) and 5.5 d(3-9), respectively(P<0.05). The pathologic examination showed negative surgical margin in all cases of renal cell carcinoma. The patients were followed up for 4 months to 32 months. Neither distant nor local recurrences were observed, and the renal function was normal in all cases. Urinary leakage oc-curred in 1 patient of each group. Conclusions CTA could be an effective method in evaluating the supply blood vessels of kidney before LNSS. And it can shorten the operation time and warm ischemia time, reduce the blood loss, as the result of helping the operaters deal with renal artery more quickly and accurately.

Objective By using percutaneous endovascular microcatheter technique to establish an animal model of coronary microembolization in mini-swine which is suitable for long-term observation.Methods Coronary microembolization was established in 10 mini-swine by infusing 15 × 10~4 microspheres (φ45μm) selectively into the left anterior descending artery (n = 10). Coronary flow reserve (CFR) was measured by Doppler wire and left ventricular eject fraction (EF) was assessed by echocardiography.Hematoxylin and eosin (HE) staining and nitroblue tetrazolium (NBT) dye were used to demonstrate the presence of microembolization after the procedure of coronary microembolization. The ultra-structures of cardiomyocyte were observed by transmission electron microscopy (TEM). Before sacrifice, the CFR measurement and coronary angiography were performed again in survival animals. Results The coronary microvascular integrity (CFR < 2.0) and left ventricular function (EF < 50% ) were damaged by coronary microembolization. One month after the procedure, all the 10 animals survived and were able to receive the angiography and CFR measurement again. HE staining and NBT dye could demonstrate the presence of microembolization. The edema and fibrosis of cardiomyocytes could be revealed with TEM. Conclusion The animal model of coronary microembolization can be established in mini-swine by using percutaneous endovascular microcatheter technique. The model is suitable for long-term observation, the preparation is technically-simple and minimally-invasive with very low mortality. Therefore, this kind of animal model is an ideal experimental form for studying the mechanism of coronary microembolization.

Objective To discuss the techniques and clinical efficacy of laparoscopic partial cystectomy with bilateral pelvic lymphadenectomy for urachal adenocarcinoma. Methods From July 2006 to April 2008, 4 patients with urachal adenocarcinoma were managed by the laparoscopic procedure. Three patients were male, the other one was female, with a median age of 51 (range 42 to 66)years. The mean size of tumors was 3.4(rang 1.9 to 5.4)cm in diameter. Three of them were diagnosed as mucinous adenocarcinoma, the other one was adenocarcinoma. There was 1 patient at stage Ⅱ , and the other three as stage Ⅲ according to Sheldon Stage. Four patients were performed by transperitoneal approach. The boundaries of resection were similar to the open surgery, including resection of the tumor with normal margins, the peritoneum lateral to the two medial unbilical ligaments,the posterior sheath of the rectus muscle and the muscle fibers of the rectus muscle below it, and bilateral pelvic lymphanodes. Results The procedure was successfully in all 4 patients, with a mean operative time of 220(range 150 to 350)min, a mean estimated blood loss of 180 (range 120 to 290)ml.No significant intraoperative or postoperative complications occurred, except for an inferior epigastric artery injury in 1 case. The mean postoperative in-dwelling urinary catheter time was 6 (range 5 to 7)d, and the mean postoperative hospital stay was 6 (range 5 to 8)d. All 36 resected lymph nodes (range 8 to 11) were negative. At a median follow-up of 25(range 15 to 36) months, there was no evidence of recurrent disease by radiologic or cystoscopic evaluation. ConclusionLaparoscopic partial cystectomy and bilateral extended pelvic lymphadenectomy in selected patients with urachal tumors could be a safe, feasible, minimally invasive procedure.

Objective To investigate the prevalence of vitamin D deficiency and its associated metabolic risk factors in the healthy adults in Gansu Province, a western province of China.Methods Through stratified, multistage probability population sampling, 11 300 adults (4 740 men and 6 417 women aged 18-79 years) who had been living in their current residence for at least 5 years were arbitrarily chosen from seven representative geographical cities in Gansu Province from June 2014 to December 2015.The data obtained from a questionnaire-based survey, anthropometric and biochemical assessments, and areal bone mineral density measurement were collected and analyzed.Results The overall mean serum 25(OH)D was (16.07±9.40) μg/L.The prevalence of severe deficiency (<10 μg/L), deficiency (≥10 μg/L and <20 μg/L), insufficiency (≥20 μg/Land <30 μg/L), and sufficiency (≥30 μg/L) among these adults were 17.3％, 64.6％, 11.8％, and 6.3％, respectively.The significant predictors of vitamin D deficiency included younger age and female sex(P<0.05), whereas sun exposure, physical activity, and calcium supplementation were protective (P<0.05).Serum 25(OH)D was inversely associated with parathyroid hormone (r=-0.279, P<0.001) and positively associated with serum calcium (r=0.239, P<0.001), serum phosphorus (r=0.090, P=0.018), low-density lipoprotein cholesterol(r=0.100, P=0.008), and body mass index (r=0.093, P=0.014).The 25(OH)D concentration showed no significant correlation with bone mineral density (P>0.05).Conclusions Vitamin D deficiency is highly prevalent among adult populations in Gansu, northwestern of China, and is largely attributed to younger age and female sex.The cumulative effects of factors including sun exposure, physical activity, and calcium supplementation play an influential role in maintaining vitamin D levels.

AIM: To isolate, purify and differentiate endothelial progenitor cells from cord blood in vitro and to study their biological characteristics. METHODS: CD133~+ cells were selected from fresh cord blood mononuclear cells (MNC) by magnetic activated cell-sorting system (MACS). EPC was studied by flow cytometry, immunocytochemistry and immunofluorescence staining. Isolated cells were cultured in IMDM medium supplemented with or without VEGF, bFGF, SCF. RESULTS: The percentage of CD133~+ cells of cord blood MNC was (1.41?1.14)%, and purity was 75%-85% (FACS method). CD133~+ cells were grown on fibronectin-coated chamber slides in the presence of VEGF, bFGF, SCF. Within 1-2 hours of culture cells became adherent. On day 7-10, the adherent cells displayed a typical "cobblestone" morphology. After 14 days of culture, the adherent cells revealed a heterogeneous cell population, comprising small-sized round cells, spindle-like cells and formed tube-like structure. Weibel-Palade bodies were shown on the transmission electron microscopy photomicrographs. Compared with the (original,) cell markers CD133 and CD34 decreased significantly (77.0%?3.3% to 1.6%?2.2% and 93.1%?4.7% to 37.4%?4.9%, P

Objective To assess the prognosis of patients with bladder neck (BN) involvement in radical prostatectomy specimens and compare it with patients with seminal vesicle invasion (SVI).Methods From January 2002 to December 2008,42 patients with prostate carcinoma who underwent radical prostatectomy were analyzed retrospectively.The mean age was 70.3 years (range 59-78 years).The patients were divided into two groups according to 2002 TNM system:pT4a group (bladder neck invasion) with 17 cases and pT3b group (unilateral or bilateral seminal vesicle invasion)with 25 cases.Postoperative outpatients were followed-up on a regular basis,two consecutive postoperative serums PSA greater than 0.2 μg/L defined as a biochemical recurrence.The postoperative follow-up to the time of biochemical recurrence or follow up to the deadline without biochemical recurrence were defined as biochemical recurrence free survival time.Results All patients were followed up for 50-122 months.The patients with BN involvement 17.6％ (3/17) had biochemical recurrence,whereas seminal vesicle invasion 24.0％ (6/25),there was no statistical significance between the two groups (P＞0.05).Conclusions Postoperative adjuvant therapies can improve the biochemical recurrence free survival.In this study,the prognosis of pT4a and pT3b group is similar.A conceivable down-staging of BN involvement in the TNM staging system should be considered.

ObjectiveTo evaluate the clinical effecacy and safety of transurethral enucleation of submucosal bladder leiomyoma.MethodsAnalyze retrospectively the clinical data of 6 patients (2 male,4 female) of submucosal bladder leiomyoma. The mean age was 59 years (range 32- 78). The clinical manifestations included dysuresia in 3 cases, irritative bladder in 1 case, gross hematuria in 1 caes, and no clinical symptoms in 1 case. The mean course was 23 months (range 1 week-4 years). All the bladder tumors were indicated by ultrasonography, CT scan and cystoscopy, while 4 cases of bladder filling defects were showed by intravenous urogram. Before the tranaurethral enucleation of the bladder tumor, a deep needle biopsy and pathological examination were carried out to confirm the diagnosis of bladder leiomyoma. For the relatively small lateral leiomyomas, holium laser enucleation was carried out, while for the big ones, resectoscope enucleation was used to remove the mass. Biopsys were performed after complete removal of the tumor.ResultsTransurethral enucleation of 6 cases of submucosal bladder tumors were successful without any postoperative complication. All patients regained normal urination, and lower urinary tract irritation was relieved obviously and hematuria disappeared. No tumor recurrence or metastasis was found during the follow-up of 4 to 158 months.ConclusionsCystoscopy and transurethral biopsy are the most important methods for the diagnosis of submucosal bladder leiomyoma. The transurethral enucleation is a feasible and safe surgical technique for such patients with excellent results.

ObjectiveTo observe the clinical characteristic and treatment response of the Gleason 5 + 5 prostate cancer and to summarize the prognostic factor of such patients. MethodsFrom January 2005 to May 2010,21 cases of Gleason 5 + 5 prostate cancer were enrolled in this study.The average age was 73 years and the pre-treatment PSA was 60.8 ng/ml.Six of the 21 cases had a PSA level ≤20 ng/ml.The maximal androgen deprivation therapy (ADT) in the form of castration and Casodex or fluctamide was carried out once the diagnosis was made.Bone metastasis was revealed in 16 cases.PSA levels were tested at the 4th month and then every 3 months after the castration.The observation endpoint was 6 months.Extracorporeal beam irradiation or the systemic chemotherapy was carried out if the PSA did not drop to a normal range after 6 months. ResultsIn the study group with initial PSA ＞ 20 ng/ml,the PSA did not drop to normal range in 5 cases; of which 4 had bone metastasis.With systemic chemotherapy,3 cases died within 1 year.Another case,with localized disease,received an extracorporeal beam irradiation and died within 1 year due to progression of the tumor.Among the remaining 10 cases sensitive to total androgen blockage,7 survived through the end of the study period.One case had PSA recurrence in month seven after the initial therapy and died at month 19.One case had PSA recurrence at month 22 and died at month 36.The patient had his PSA augmented at month 24 after total androgen blockage and the patient passed away 4 months later.Of the 6 cases who had an initial PSA ≤20 ng/ml 4 died within one year. ConclusionsGleason 5 +5 prostate cancer is relatively resistant to ADT with a worse prognosis,especially for the cases with a nadir PSA ＞4 ng/ml after 6 months’ ADT.The patients who had an initial PSA ≤20 ng/ml might have an unsatisfactory clinical outcome.