Objective To establish a polymerase chain reaction method for the detection of the rcp virulence gene of L. pneumophila. Methods Sixteen strains of Legionella pneumophila, L. dumiffii, L. bozemanii and L. Longbeachae isolated from the cooling towers of the centralized air-conditioning systems in Tianjin form 2005 to 2007 were detected by polymerase chain reaction method using L. pneumophila rcp virulence gene-specific primer according to GenBank published nucleotide sequence. Results The 900 bp rcp genetic fragment was amplified in three strains of L. pneumophila, while others not. Conclusion The rcp virulence gene-based polymerase chain reaction method for the detection of L. pneumophila has been successfully established and is the foundation for the studies of Legionella virulence.

Objective To evaluate if the flexible ureteroscopy could treat stones located in lower calyx with the infundibulopelvic angle (IPA) less than 30°.Methods Thirty-six patients with inferior caliceal calculi on whom flexible ureteroscopic procedures were performed between November 2009 and June 2015 were reviewed.The mean age of the patients was 52.1 years (34-71),with the mean stone diameter of (1.5 ± 0.8) cm (1.2-2.6 cm).IPA was smaller than 30° in all 36 cases,which confirmed by CTU examination.IPA was less than 10° in 15 patients,between 11 ° and 20° in 13 patients and between 21 ° and 30° in 8 patients.Results The success rate was 63.9％ (23/36 patients) in patients with IPA smaller than 30° after first session of procedure,and the stone free rate reached 100％ after the second session of procedure.The mean operation duration was (95.5 ± 31.4) min(51-127 min).The mean hospital stay after operation was(4.1 ± 1.2)days (3-5 days).No major complications were recorded and no patients needed to convert to open surgery.Double J tube was removed after 4 weeks postoperatively.Patients were followed up for 4-12 months,during which ultrasound and CT scan were used for stone detection.Conclusions The small IPA (＜30°) negatively affected the SFR in the first session operation.However,its negative effect was solvable by using modern endoscopes.A complete stone clearance was achievable even in case of unfavorable anatomic conditions in experienced hand.

Object To select the endophytic fungi which produce vincristine by isolating fungi from the leaves of Catharanthus roseus (L.) G. Don. Methods The endophytic fungi were isolated from the leaves of C. roseus and the zymotic extracts were analyzed by TLC and HPLC. Results An endophytic fungus which is Mycelia sterilia 97CY_3 can produce vincristine. The content of vincristine in the fungus was determined as 0.205 ?g/L by HPLC. Conclusion Some endophytic fungi isolated from C. roseus can produce the anticancer substance vincristine which is the same as that of host plant producing.

Objective To study the immunoregulatory effects of bone marrow mesenchymal stem cells (MSC) on allogeneic peripheral B lymphocytes in vitro. Methods MSCs were isolated and cultured from bone marrow by gradient centrifugation. Mononuclear cells were isolated routinely from peripheral blood, then monocytes were eliminated by L-leucine methy ester method. Remained T lymphocytes were eliminated by AET-SRBC rosette method. The action of MSCs and its supernatant on B lymphocytes proliferation in the presence of anti-human IgM goat antibodies (anti-IgM) was investigated by MTT. The IgG, IgM in the supernatant were detected by ELISA. The percent of apoptosis B lymphocytes, co-cultured with MSCs for 24 or 48h, was assayed by FACS. Results MSCs and its supernatant inhibited B lymphocytes proliferation and Ig secretion. The inhibitory effect depended on the amount of MSCs and condition of its supernatant. The date of FACS indicated that the apoptosis ratio of B lymphocytes, co-cultured with MSCs for different times, were non-significant. The inhibitory effect of MSCs on B lymphocytes was temporary and reversible. Conclusion MSCs have immunoregulatory effects on B lymphocytes, and its mechanisms are complex, not only correlating with the concentration of MSCs but also the action between cells and the secretory cytokine of MSCs.

Objective To monitor the renal pelvic pressure and to investigate its clinical significance during retrograde flexible ureteroscopic lithotripsy (RFUL).Methods The data of renal pelvic pressure measured in 60 cases of RFUL with the mean irrigation pump speed and pressure of 30 ml/min and 30 mmHg were analyzed retrospectively.The influence factors of renal pelvic pressure and its correlation with postoperative fever were analyzed.Renal pelvic pressure was measured by baroceptor,which was connected to PHILIP-MP4 monitor IBP channel and ureteric catheter positioned in renal pelvis through a dual channel ureteral access sheath (UAS).The renal pelvic pressure data was collected and analyzed in every 2 seconds by computer.The 60 cases were divided into 3 groups according to their intra-pelvic pressure situations:normal pressure group(NP,IPPmax ≤30 mmHg),high pressure group(HP,IPPmax＞30 mmHg,but high pressure duration≤ 10 min),and backflow pressure group(BP,IPPmax＞30 mmHg and high pressure duration＞ 10 min).Results The baseline intra-pelvic pressure (IPP0) and max imum intra-pelvic pressure (IPPmax) were (13.2±5.6) mmHg and (95.6±2.3) mmHg respectively.IPP levels during the RFUL were significantly higher than the IPP0(P＜0.001).There were 32,17 and 11 cases in NP,HP and BP groups,respectively.There were 6 cases with fever higher than 38.5 ℃ (10％),in which there were 1 case in NP,1 case in HP group and 4 cases in BP group.The postoperative fever rate in NP,HP and BP group were 3％,6％ and 36％ respectively,which were significantly different between groups(P＜0.01).There were 12 cases with procalcitonin ＞0.1 ng/ml and 8 cases with procalcitonin ＞0.5 ng/ml,in which 2 cases in HP group and 6 cases in BP group.Conclusions RFUL would result in a temporal elevated intrapelvic pressure greater than 30 mmHg.Postoperative fever is relevant with renal perfusion pressure and perfusion time.It's necessary for the surgeons to adjust the perfusion pressure during operation.

Objective To compare the therapeutic effect of retrograde ureteroscopic Ho:YAG laser lithotripsy (UHL) and percutaneous nephrolithotripsy (PCNL) in treating patients with renal calculi of ≤3 cm.Methods From Feb.2008 to Apr.2011,a total of 109 cases (117 renal calculi in total) treated by either PCNL (50) or UHL (67) were retrospectively analyzed.Operative time,stone free rate,postoperative hospital stay,hospitalization expenses and complications were compared in the 2 groups.Results The mean stone burden of the UHL group and PCNL group was 2.5 cm (1.5-3.0) cm and 2.4 cm (1.3-3.0) cm,respectively.There was no significant difference in two groups.In UHL group,operations were performed successfully in all 67 calculi,of whom,43 patients needed combination of flexible ureteroscopy to break the stone fragments falling into the renal calices.No serious complication was recorded except postoperative fever in 3 cases.In PCNL group,all patients had been successful operated.Postoperative fever occurred in 2 cases.Obvious intraoperative and postoperative haemorrhage appeared in 1 case,and cured by selective renal artery embolization.The operation time of the UHL group and PCNL group was (117.0±36.5) min and (90.0±18.3) min respectively,and had no significant difference in two groups (P＞0.05).The stone free rate of the UHL group and PCNL group was 94％ (63/67) and 92％ (46/50) respectively,and had no significant difference in two groups (P＞0.05).Hemoglobin decline of the UHL group and PCNL group was (0.3±0.1) g/L and (20.6±8.1) g/L,and had significant difference in two groups (P＜0.05).Postoperative hospital stay of the UHL group and PCNL group was (5.0±2.4) day and (7.0±3.7) day,and had statistical difference in two groups (P＜ 0.05).Hospitalization expenses of the UHL group and PCNL group was (15 477.0±754.3) RMB and (27 453.0± 1763.5) RMB,and had statistical difference in two groups (P＜0.05).Conclusions Although UHL and PCNL have similar curative effect in treating renal stones of less than 3 cm,UHL have the advantages of lcss trauma and complication,lower expenses and fewer hospital stay.In treating renal stones of smaller than 3 cm,especially in the contraindication of PCNL,UHL may be a better choice.

The cloning and identification of frc gene from Oxalobacter formigenes in the intestines of Chinese people were conducted. The genomic DNA of Oxalobacter formigenes was extracted. frc gene fragment was amplified by polymerase chain reaction (PCR) and linked with pEGFP-C1. The recombinant plasmid was designated pEGFP-frc and was identified by restriction-enzyme digestion and sequencing. Human embryo kidney 293 cells were transfected with pEGFP-frc, then RT-PCR and Western blotting were performed to detect the expression of frc gene. The length of frc gene was found to be 1287 bp, and the homology of nucleotides and amino-acid residue with the sequence in GenBank was 95.88% and 99.07%. Bright green fluorescent light could be observed in 293 cells transfected with the pEGFP-frc. frc mRNA and fusion protein FCoAT-EGFP were detected in the cells. It is concluded that frc gene cloned from the Oxalobacter formigenes in the intestines of Chinese people can be expressed in eucaryotic 293 cells and keep its enzyme activity.

ObjectiveTo evaluate term efficacy of focal cryoablation for localized single lesion prostate cancer and the protection of erectile function.MethodsFrom June 2008 to December 2009, 12 patients with localized single lesion prostate cancer were treated with focal cryoablation. All diagnoses were confirmed by transrectal ultrasound-guided prostatic biopsy. The mean Gleason score was ≤ 7; three cases had a Gleason score of 5, five cases a score of 6, and four cases a score of 7. The average preoperative value of PSA was 9.7 ng/ml ( 4.2 - 14.9 ng/ml ). The PSA level was obtained every three months for the first year and then every six months thereafter. Patients with a PSA nadir greater than 1.0 ng/ml or if the PSA nadir increased more than 2 ng/ml underwent repeat biopsy to exclude tumor recurrence.ResultsThe mean operative time was (82 ±26) minutes. The mean postoperative hospital stay was (5 ± 2) days. After catheter removal, all 12 cases had good continence and 10 had erectile function before surgery. Of the 10 patients, eight had erectile function after surgery. The follow - up ranged from 12 to 30 months ( mean 23 ).The lowest average serum PSA nadir was 2.2 ng/ml, nine patients had a minimum PSA nadir of less than 1.0 ng/ml and three had a minimum PSA nadir of more than 1.0 ng/ml. Four patients underwent repeat biopsy after abnormal PSA, and prostate cancer was detected in the contralateral gland in one patient.ConclusionsUltrasound-guided transperineal focal cryoablation is a safe and effective treatment with few complications for localized single lesion prostate cancer. Additional studies with longer follow-up are necessary for long-term efficacy.

OBJECTIVETo study the stomach role in exogenous oxalate absorption.

METHODSThe kinetic variation of urinary oxalate excretion (mg/min) in 10 healthy adults and 8 patients who underwent total gastrectomy was investigated before and after an oral spinach oxalate load. The bioavailability of the oxalate load in the healthy adults was calculated and compared with that in the patients.

RESULTSThe oxalate content in the oral spinach load was 2567-2670 mg. The urinary oxalate excretion (mg/min) in the 10 healthy adults increased significantly 20 minutes after loading (this increase was compared against their basic oxalate excretion level of 0.0331 +/- 0.0203). Further observations after loading include: a first peak of oxalate excretion 40 minutes after loading; an oxalate excretion level double that of the basic level after 60 minutes (0.0732 +/- 0.0294) and a second peak appearing at 3 hours (P < 0.01). A "first peak" (0.063% +/- 0.062%) was not in any of the patients who underwent a total gastrectomy. Furthermore, a bioavailability of oxalate, which was 50% lower than that in the healthy subjects, appeared 60 minutes after loading (0.098% +/- 0.071%, P < 0.01).

CONCLUSIONSThe stomach is a powerful oxalate absorption organ under normal physiological conditions. Further investigation on the relationship between stomach dysfunction and urinary calcium oxalate formation is needed.

Absorption ; Adult ; Female ; Humans ; Male ; Middle Aged ; Oxalates ; pharmacokinetics ; Spinacia oleracea ; metabolism ; Stomach ; metabolism