The inter-species differences of estradiol metabolism were investigated in human, Beagle dog and rat liver microsomes by comparing enzyme kinetics of parent drug and the formation of its major metabolites. The incubation systems of estradiol with liver microsomes of the three species were optimized in terms of estradiol concentration, microsomal protein content and incubation time. The concentrations of estradiol and its metabolites were measured by LC-MS/MS method. The t1/2, CLint, CLh, Km and Vmax of estradiol incubated with male human liver microsomes were 40.02 +/- 8.32 min, 41.39 +/- 6.57 mL x min(-1) x kg(-1), 13.81 +/- 12.36 mL x min(-1) x kg(-1), 26.8 +/- 6.99 micromol x L(-1) and 0.75 +/- 0.92 micromol x L(-1) x min(-1), respectively. The corresponding parameters of female human were 44.71 +/- 10.21 min, 29.85 +/- 8.97 mL x min(-1) x kg(-1), 0.01 +/- 0.68 mL x min(-1) x kg(-1), 44.2 +/- 7.73 micromol x L(-1) and 1.27 +/- 4.41 micromol x L(-1) x min(-1), that of male dog were 21 +/- 7.33 min, 165.53 +/- 29.33 mL x min(-1) xkg(-1), 26.01 +/- 8.39 mL x min(-1) x kg(-1), 19.5 +/- 7.34 micromol x L(-1) and 1.6 +/- 0.65 micromol x L(-1) x min(-1), that of female dog were 25.5 +/- 5.32 min, 135.11 +/- 42.34 mL x min(-1) x kg(-1), 0.24 +/- 3.18 mL x min(-1) x kg(-1), 8.33 +/- 6.32 micromol x L(-1) and 0.51 +/- 2.15 micromol x L(-1) x min(-1), that of male rat were 5.11 +/- 3.84 min, 485.63 +/- 36.52 mL x min(-1) x kg(-1), 49.57 +/- 15.29 mL x min(-1) x kg(-1), 62 +/- 13.74 micromol x L(-1) and 19.16 +/- 9.67 micromol x L(-1) x min(-1), and that of female rat were 7.0 +/- 3.69 min, 354.82 +/- 33.33 mL x min(-1) x kg(-1), 8.04 +/- 3.23 mL x min(-1) x kg(-1), 35.38 +/- 7.65 micromol x L(-1) and 8.39 +/- 4.91 micromol x L(-1) min(-1), respectively. There were nine metabolites detected from all the three species, but the relative amounts of the metabolites generated were different in three species. The results indicted that the major phase I metabolic pathway of estradiol was similar in the liver microsomes from all the three species. However, the inter-species differences were found in the view of relative amounts of the metabolites as well as the metabolic characteristics of estradiol in liver microsomal incubation.

Adult ; Asthma ; metabolism ; physiopathology ; Basic-Leucine Zipper Transcription Factors ; genetics ; physiology ; Fanconi Anemia Complementation Group Proteins ; genetics ; physiology ; Female ; Glutamate-Cysteine Ligase ; metabolism ; Humans ; Inflammation ; metabolism ; pathology ; Male ; NF-E2-Related Factor 2 ; genetics ; physiology ; RNA, Messenger ; genetics ; metabolism ; Sputum ; cytology

Objective To evaluate the effect of hydrogen-rich saline on the expression of phosphor-p38 mitogen-activated protein kinase (p-p38MAPK) during cerebral ischemia-reperfusion (I/R) in rats.Methods Seventy-two adult male Sprague-Dawley rats,weighing 220-250 g,were randomly divided into 3 groups (n =20 each) using a random number table:sham operation group (group S),I/R group and hydrogen-rich saline group (group I/RH).Cerebral ischemia was induced in chloral hydrate-anesthetized rats by 2 h middle cerebral artery occlusion in I/R and I/RH groups.The artery was only exposed but not occluded in group S.At 3 days before operation and immediately after onset of reperfusion,hydrogen-rich saline (0.6 mmol/L) 10 ml/kg was intraperitoneally injected in group I/RH,while the equal volume of normal saline was given in S and I/R groups.Neurological deficits were blindly assessed and scored at the end of 24 h reperfusion.The animals were then sacrificed,and brains were removed for microscopic examination and for determination of the cerebral infarct size (by TTC),brain water content,cell apoptosis (by TUNEL),and expression of p38MAPk and phosphor-p38MAPK (p-p38MAPK) (by immunohistochemistry and Western blot).Apoptosis index was calculated.Results Compared with group S,neurological deficit score,apoptosis index,brain water content and cerebral infarct size were significantly increased,and the expression of p38MAPK and p-p38MAPK was up-regulated in I/R and I/RH groups.Compared with group I/R,neurological deficit score,apoptosis index,brain water content and cerebral infarct size were significantly decreased,and the expression of p38MAPK and p-p38MAPK was down-regulated in group I/RH.The pathological changes of cerebral tissues were significantly attenuated in group I/RH as compared with group I/R.Conclusion Hydrogen-rich saline can reduce cell apoptosis through inhibiting p-p38MAPK expression,thus attenuating cerebral I/R injury in rats.

Objective To evaluate the effect of hydrogen on the activation of caspase-3 in brain tissues during cerebral ischemia-reperfusion (I/R) in rats.Methods Thirty-six healthy adult male Sprague-Dawley rats,weighing 220-250 g,were divided into 3 groups (n=12 each) using a random number table:sham operation (group S),I/R group and hydrogen group (group H).Cerebral ischemia was induced by occlusion of the middle cerebral artery followed by reperfusion in I/R and H groups.In group H,hydrogen-rich saline 5 ml/kg (0.6 mmol/L) was injected intraperitoneally at 3 days before establishment of the model and immediately after the onset of reperfusion.At 24 h of reperfusion,the rats were sacrificed,and hippocampal tissues were obtained for determination of neuroapoptosis (by TUNEL),apoptotic neuron count and expression of activated caspase-3 (by Western blot).The brain tissues in the ischemic area were obtained and stained with haematoxylin and eosin for examination of the pathological changes.Results Compared with group S,the expression of activated caspase-3 was significantly up-regulated,and the apoptotic neuron count was increased in I/R and H groups (P<0.05).Compared with group I/R,the expression of activated caspase-3 was significantly down-regulated,the apoptotic neuron count was decreased (P<0.05),and the pathological changes of brain tissues were significantly reduced in group H.Conclusion The mechanism by which hydrogen inhibits neuroapoptosis during cerebral I/R is probably related to inhibited activation of caspase-3 in brain tissues of rats.

OBJECTIVETo explore the therapeutic effect of combination of Zhuanyindan (ZYD, a self-made Chinese herbal preparation) and hormone in treating male infertility with positive antisperm antibody and its influence on nitric oxide (NO) level.

METHODSEighty-two patients were randomly divided (according to the digital list) into the WM group (n = 20, treated with prednisone), the TCM group (n = 28, treated with ZYD) and the ICWM group (n = 34, treated with prednisone plus ZYD). The clinical effect, negative converting rate of antisperm antibody, changes of NO level in semen and various parameters of sperm motion before and after treatment were observed.

RESULTSThe total effective rate in the ICWM group was 88.2%, that in the TCM group 75.0% and in the WM group 65.0%. Significant difference was seen in the ICWM and TCM group before and after treatment in NO level, sperm motion parameters, including linear motion speed, linearity, propulsion, whip frequency, sperm vitality and mean moving angle, and quality of semen (P < 0.05 or P < 0.01). In the WM group, significant difference in comparison before and after treatment was seen in NO level, propulsion, whip frequency, mean moving angle and quality of semen, including vitality and survival rate (P < 0.01).

CONCLUSIONCombination of Chinese herbs and hormone could lower the NO level in semen and improve the quality of sperm.

Adult ; Autoantibodies ; metabolism ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Infertility, Male ; drug therapy ; etiology ; immunology ; Male ; Nitric Oxide ; metabolism ; Phytotherapy ; Prednisone ; therapeutic use ; Semen ; metabolism ; Sperm Motility ; drug effects ; Spermatozoa ; immunology

ObjectiveTo summarize the initial experience of transumbilical laparoendoscopic single-site surgery of urology.MethodsFrom February 2010 to March 2011,21 patients underwent laparoendoscopic single-site surgery using transumbilical single-site and common surgical instruments of laparoendoscopic.Nine patients underwent single-site laparoscopic ureterolithotomy,5 underwent transumbilical single-site laparoscopic ureteral stricture resection and anastomosis,5 underwent transumbilical single-site laparoscopic renalcyst unroofing and 2 had a nephrectomy.All of the cases were definitely diagnosed.A single umbilical incision of 1.5 cm to 2.5 cm was made for Triport.The procedures were performed according to the methods used in classical laparoscope methods using general instruments.ResultsAll the operations were successfully completed without conversion to open surgery.The mean operative time of ureterolithotomy was 143 (120-230) min,the mean operative time of ureteral stricture resection and anastomosis was 157 (120 -180) min,the mean operative time of unroofing of renal cysts was 110 (95 -132) min,and the operative time of the nephrectomy was from 95 to 120 min.The intestinal tract function recovered within 1 -2 d,the drainage tube was removed within 2 -3 d and the postoperative hospitalization duration was 4 -7 d.The symptoms were reduced or disappeared and no major intraoperative or postoperative complications occurred within 4 - 6 months.Conclusions Transumbilical laparoendoscopic single-site surgery represents a safe and feasible operation for urologic patients.With more clinical practice,laparoendoscopic single-site surgery could be generally applied.

Objective To explore the methylation status of hMLH1 and hMSH2 promoter region in the epithelial ovarian cancer and its role in oncogenesis.Methods Methylation status of hMLH1 and hMSH2 promoter region was assayed in 20 normal ovarian tissues,25 benign epithelial tumor,56 malignant epithelial tumor and cell lines SKOV3,3AO by methylation-specific PCR (MSP).SKOV3 and 3AO were analyzed before and after 5-aza-2′-deoxycytidine (5-Aza-CdR) treatment.In addition,an alterations of mRNA expression of hMLH1 and hMSH2 was observed by reverse transcription polymerase chain reaction (PT-PCR).Results No methylation of hMLH1 and hMSH2 promoter was found in normal ovarian tissues. CPG islands methylation of hMLH1 and hMSH2 was observed in 4% (1/25),8% (2/25) respectively in benign epithelial tumor,30.4% (17/56),51.8% (29/56) respectively in malignant epithelial tumor. Methylation status in promoter showed obvious correlation with pathological grade and lymph node metastasis (P

Objective To detect the expression of urinary nuclear matrix protein 22(NMP22) and cytokeratin 20(CK20) mRNA in the monitoring of bladder tumor recurrence ,and to explore the clinical value of combined detection of urinary NMP22 and CK20 mRNA in the monitoring of bladder tumor recurrence .Methods Enzyme‐linked immunosorbent assay(ELISA) and nested reverse transcriptase polymerase chain reaction(Nested RT‐PCR) were used to detect the expression of NMP22 and CK20 mRNA in the u‐rine of 46 patients with recurrent bladder cancer tumor (recurrent group) ,66 patients without recurrent tumor (no‐recurrent group) and 40 healthy volunteers(control group) respectively .Results The expression of NMP22 and CK20 mRNA in the urine of control group were negative .There was statistically significant difference between the positive expression rate of NMP 22 and CK20 mRNA in the urines of the recurrent group[78 .3% (36/46) ,80 .4% (37/46))] and that of the no‐recurrent group[6 .1% (4/62) , 6 .1% (4/62)] and the control group respectively (P<0 .01) .There was no statistically significant difference between the no‐recur‐rent group and the control group(P>0 .05) .The expression level of NMP22 in recurrent tumor increased obviously along with the progression of the pathological grade and clinical classification of tumor (P<0 .05) .The positive rate of CK20 mRNA expression in the recurrent group increased obviously along with the increase of the tumor number (P<0 .05) .The sensitivity and specificity of NMP22 for monitoring bladder cancer recurrence were 78 .3% and 93 .9% ,of CK20 mRNA were 80 .4% and 93 .9% .Combined de‐tection of both sensitivity and specificity were 95 .7% and 92 .4% .Statistical significance was observed between combined sensitivi‐ty and that of single detection(P<0 .05) .No significant difference was observed between combined specificity and that of single de‐tection(P>0 .05) .Conclusion The positive expression of CK20 mRNA may also indicate tumor multiplicity .Combined detection of NM P22 and CK20 mRNA significantly increases the sensitivity and specificity in clinical detection .

Objective To assess the safety and efficacy of abdominal radical nephrectomy and systematic lymph node dissection for treatment of renal carcinoma. Methods A total of 136 patients underwent radical nephrectomy and regional clearance of lymph nodes from July 2004 to June 2008.There were 92 males and 44 females in the study group.Ages ranged from 23 to 81 years,with a mean age of 54 years.The mean tumor diameter was 55 mm (range,15-170 mm).The tumor size detected by CT and MRI was consistent with that detected by B-ultrasound,98 were stage Ⅰ,13stage Ⅱ,12 stage Ⅲ,and 2 stage Ⅳ. Results All 136 cases underwent radical nephrectomy with retroperitoneal lymphadenectomy.All operations were successful without any major complication.The operative time was 90 to 180 min,with an average of 120 min,and blood loss was 20-400 ml,with an average of 50 ml.The pathological diagnoses were as follows: renal cell carcinoma 123 cases (90%), papillary renal cell carcinoma six cases(4%),chromophobic two cases(1.4%),oncocytoma two cases(1.4%),collecting duct two(1.4%),and others three cases(2.2%).Eight cases reported positive lymph nodes.Of the 136 cases,92 cases were T1 N0 M0,11 were T2 N0 M0,10 were T3 N0 M0,eight were T3 N1 M0 and two were T1 N0 M1.Ninety-five cases (70%) were followed-up at six to 40 months (mean,20 months).The one year and three year survival rates were 96% (91/95) and 86% (82/95),respectively.Conclusions Radical nephrectomy with systematic lymph dissection has advantages of accurate staging,effective resecting of renal tumors and preventing recurrence.Radical nephrectomy is an effective method for the treatment of renal carcinoma.

Objective To retrospectively analyze the prognostic factors for locoregionally recurrent early?stage extranodal nasal?type natural killer/T?cell lymphoma ( NKTCL) . Methods A total of 56 patients with early?stage extranodal nasal?type NKTCL, who had locoregional recurrence after initial treatment and then received salvage treatment from 1995 to 2014, were enrolled as subjects. The effects of salvage treatment on the overall survival ( OS) rate were analyzed after initial treatment and recurrence. Univariate and multivariate prognostic analyses were performed on the OS rate after recurrence. Results The median follow?up time was 35. 9 months after initial treatment and 14. 8 months after recurrence. The 3?year OS rate was 73% after initial treatment and 58% after recurrence. Compared with chemotherapy alone, radiotherapy?containing salvage treatment significantly improved the OS rates after initial treatment and recurrence ( P=0. 040, 0. 009 ) , and re?irradiation also significantly improved the OS rates after initial treatment and recurrence (P=0. 018, 0. 019). Most (84%) of the acute and late adverse reactions after re?irradiation were grade 1?2 ones. The univariate and multivariate analyses showed that the Karnofsky Performance Status score, radiotherapy in initial treatment, and radiotherapy in salvage treatment were influencing factors for the OS rate after recurrence. Conclusions Radiotherapy achieves improved survival and tolerable toxicities, making it indispensable in the treatment of locoregionally recurrent extranodal nasal?type NKTCL.