Objective To evaluate the methylation status of prostate cancer NDRG1 gene promoter region,and to explore the influence of methylation inhibitor 5-azacytidine on NDRG1 gene's mRNA expression in prostate cancer cells and its effects on cell proliferation.Methods Bisulfite-sequencing PCR (BSP) were used to detect the NDRG1 gene promoter methylation status in prostate cancer and BPH tissue,prostate cancer cell lines (PC3,22RV1,LNCaP and DU145) and human normal prostate cell line's RWPE-1.After 10 μmol/L 5-azacytidine were used on LNCaP and DU145 cells for 72 h,5-azacytidine's influence on cell proliferation was analyzed with MTT,two prostate cancer cell lines NDRG1 mRNA expressions were detected with RT-PCR.Results The methylation rates of NDRG1 gene in prostate cancer cell lines PC-3,22RV1,LNCaP and DU145 were (24.8 ± 3.3) ％,(36.2 ± 2.5) ％,(48.6 ± 2.8) ％ and (69.5 ± 1.7) ％,respectively.Methylation rate of Human normal prostate cell lines RWPE-1 was (4.8 ± 4.5) ％;prostate carcinoma was (48.6 ± 5.3) ％,BPH tissue was (4.3 ± 2.1) ％.The differences between groups were statistically significant.After 10 μmol/L 5-azacytidine added on LNCaP and DU145 cells for 72 h,NDRG1 gene demethylation occurred in both cells,its mRNA expression enhanced 8-9 times compared with previous and its cell growth was inhibited (P ＜ 0.05).Conclusions NDRG1 gene promoter region's hypermethylation is one of the reasons of its aberrant expression in prostate cancer.5-azacytidine can reverse NDRG1 gene promoter methylation status,regulate the expression of the gene and can inhibit prostate cancer cell proliferation.

Objective To observe liver functional lesion caused by combination chemotherapy containing or not containing oxaliplatin. Methods Data from 42 patients with liver functional lesion caused by chemotherapy between March 2005 and October 2007 were analyzed. All patients were diagnosed through histology or cytology detection and received chemotherapy only. Different drugs were. admitted,based on different tumors. Before chemotherapy, each patient had normal liver function without liver lesions such as liver metastasis, Hepatitis B and C, hepatic cirrhosis, etc. Furthermore, 22 received FOLFOX-4 in containing oxaliplatin group while the remaining 20 received chemotherapy excluding oxaliplatin. When liver functional lesion without the influence of any liver protectant was first observed, ALT, AST, TBIL, DBIL, IBIL, ALP, GGT and the WHO criteria of liver toxicity were analyzed. T test and Wilcoxon rank sum test were used for data analysis. Results All together 90 cycles, median 2.14 cycles, were given. According to WHO criteria of liver toxicity, 13 cases were in grade O, 21 in grade Ⅰ, 7 in grade Ⅱ, and 1 in grade Ⅲ. ALT and AST were significantly high after chemotherapy(P <0.05). Moreover, ALT and AST were significantly higher in containing oxaliplatin group than non oxaliplatin group after chemotherapy(P <0.05). Chemotherapy had no influence on bilirubin. The population distribution of accumulative chemotherapy cycles and WHO criteria of liver toxicity was similar between two groups. Conclusion Before the intervention of liver protectant, combination chemotherapy containing oxaliplatin is more likely to have liver functional lesion than other chemotherapy without oxaliplatin. It mainly presents an increase in transaminase.

OBJECTIVETo observe the effect of Chinese herbal therapy on T-lymphocyte subsets in patients with IgA nephropathy (IgAN).

METHODSTotally 36 inpatients and outpatients at Department of Nephropathy, Xiyuan Hospital, China Academy of Chinese Medical Sciences, from June 2011 to June 2013 were recruited in the treatment group, while 20 volunteers were recruited as the healthy control group. Patients in the IgAN group only took Chinese herbal decoctions by syndrome typing for 3 months (except those accompanied with hypertension additionally took antihypertensive agents such as angiotensin-converting enzyme inhibitor and/or dihydropyridines calcium antagonist). No intervention was performed in the healthy control group. The values of Th1, Th2, and CD4+ CD25+ Treg, and red blood cell number in urine were detected using flow cytometry before and after treatment. 24 h urine protein was detected using inmmunoturbidimetry.

RESULTSCompared with the healthy control group, the CD4+ CD25+ Treg level obviously decreased in the IgAN group, showing statistical difference (P < 0.01). In the IgAN group, Th1, 24 h urine protein, and urine red blood cell counts were obviously lower after treatment, showing statistical difference when compared with before treatment (all P < 0.05).

CONCLUSIONChinese herbal therapy could reduce urine erythrocyte number and 24 h urine protein of IgAN patients, and down-regulating Th1 expression might be its mechanism.

Adult ; Case-Control Studies ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Glomerulonephritis, IGA ; drug therapy ; Humans ; Male ; Middle Aged ; Phytotherapy ; T-Lymphocyte Subsets ; drug effects ; Young Adult

Objective The purpose of this study was to investigate the expression of mitochondrial motility-related gene mirol and mitochondrial energy metabolism during an acute bout of prolonged exercise. Methods CS7 BL/6 mice underwent a moderate intensity treadmill running with the slope of 0° and at the speed of 13m/min, and they were randomly divided into 5 groups:resting group(R),exercise groups running for 30min(E30),60min(E60),90min (E90) and 120min(E120),respectively. Respiratory control index and ATP synthesis activity in isolated mitochondria were detected. Skeletal muscle H_2O_2 concentration,mirol mRNA expressions were also measured. Results (1)mirol mRNA contents were significantly increased in groups E30 to E120,as compared with the group R;and mirol mRNA expression increased 32.8%, 107.6%,63.8% and 44.8% respectively in groups E30 to E120. (2)H_2O_2 contents of skeletal muscle were clearly increased in group s E30 to E120. (3)ATP synthesis activity elevated at 30 min of exercise and returned to the baseline thereafter,whereas respiratory control index without remarkable change in groups E30 to E120. Conclusion Muscular mirol mRNA expression significantly increased during 120min of exercise as compared with the resting status and thus facilitated the mitochondrial respiratory and ATP synthesis for matching the energy demand during exercise.

OBJECTIVETo investigate the expression of zinc finger E-box binding homebox 1 (ZEB1) in the prepuce of hypospadias children and its relationship to the incidence of hypospadias.

METHODSPrepuce tissues were collected from 37 children aged 6-15 months undergoing hypospadias repair and 11 age-matched controls receiving circumcision. Based on the position of the urethral meatus, the hypospadias cases were classified as severe (n = 13) and mild-moderate (n = 24). The mRNA and protein expressions of ZEB1 were determined by immunohistochemistry and RT-PCR.

RESULTSThe expression of the ZEB1 protein was remarkably higher in the severe (100% [13/13]) and mild-moderate hypospadias patients (75.0% [18/24]) than in the controls (9.1% [1/11]), with statistically significant differences between any two groups (P < 0.05). RT-PCR showed the integrated density value (IDV) of the ZEB1 mRNA expression to be (0.67 ± 0.21), (0.81 ± 0.24), and (1.55 ± 0.29) in the control, mild-moderate, and severe hypospadias patients, respectively, significantly higher in the severe hypospadias than in the control and mild-moderate hypospadias groups (P < 0.05), but with no significant difference between the latter two (P = 0.64).

CONCLUSIONThe expression of ZEB1 is significantly increased in hypospadias patients, and its upregulation is positively correlated with the severity of hypospadias, which suggests that the overexpression of ZEB1 may contribute to the development of hypospadias.

Biomarkers ; metabolism ; Case-Control Studies ; Circumcision, Male ; Foreskin ; metabolism ; Homeodomain Proteins ; genetics ; metabolism ; Humans ; Hypospadias ; classification ; etiology ; metabolism ; Immunohistochemistry ; Infant ; Male ; Penis ; RNA, Messenger ; metabolism ; Transcription Factors ; genetics ; metabolism ; Up-Regulation ; Urethra ; Zinc Finger E-box-Binding Homeobox 1

The chromatographic fingerprint was established by eluting with the mobile phase consisted of acetonitrile and 0.2% formic acid water on an Agilent TC-C18 (2) column (4.6 mm x 250 mm, 5 microm). Six chromatographic peaks were identified by HPLC-MS/MS method. Ten batches of Glycyrrhizea Radix et Rhizoma Praeparata Cum Melle were determined, and the similarity was arranged from 0.72 to 0.99. Good precision, stability and repeatability were obtained, and this study provides a reference for the quality control of Glycyrrhizea Radix et Rhizoma Praeparata Cum Melle.
China ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; Glycyrrhiza uralensis ; chemistry ; Mass Spectrometry ; Quality Control ; Rhizome ; chemistry

Objective To evaluate the contribution of puboprostatic ligament-sparing technique in urinary continence after radical retropubic prostatectomy (RRP). Methods A total of 74 men with clinically localized prostate cancer underwent RRP. Of whom, 50 patients were performed pubo-prostatic ligament-sparing technique(group A), while 24 patients were not(group B). Patients were evaluated by independent observer questionnaire to determine their urinary continence status. Results Mean patient age [(61.3±2.4) vs (60.8±2.1)years], serum prostate-specific antigen (PSA) values [14.3±1.2)ng/ml vs (14.7±1.3) ng/ml], operative time [(110.5±10.4)min vs (109.7±10.6) mini, estimated blood loss [(250.5±23.4) ml vs (253.4±22.3) ml], and positive surgical margin rate (6% vs 8%) were not significantly different between group A and B(P>0.05). The urinary con-tinence rate with the puboprostatic ligament-sparing technique at 1-year follow-up was significantly higher than that of the control group (P<0.05). Concision The puboprostatic ligament-sparing technique significantly enhances post-operative urinary continence after RRP.

Objective To evaluate the accuracy of Gallina nomogram in predicting seminal vesicle invasion (SVI) in prostate cancer.Methods From January 2009 to December 2011,89 patients with prostate carcinoma underwent open retropubic or laparoscopic radical prostatectomy.Complete data of preoperative serum prostate specific antigen (PSA),clinical stage,biopsy Gleason score,percentage of positive biopsy cores,pelvic MRI and pathological report of prostatectomy specimen were collected,and all the patients met the inclusion criteria of Gallina nomogram,2001 Partin tables and 2007 Partin tables.Postoperative pathological results were respectively compared with MRI and the incidence of SVI predicted by the three tools,and the sensitivity,specificity and accuracy of MRI in predicting SVI were calculated.The receiver operating characteristics curves were performed to test the predictive accuracy of SVI of each tool.Results The incidences of organ-confined disease,capsule invasion,SVI and lymph node metastasis were 58.4％,16.9％,22.5％,and 2.2％,respectively.The sensitivity,specificity and accuracy of MRI in predicting SVI was 45.0％ (9/20),71.0％ (49/69) and 65.2％ (58/89),respectively.The area under the curve (AUC) for SVI disease prediction of 2001 Partin tables,2007 Partin tables and Gallina nomogram was 0.712,0.711 and 0.801,respectively.Conclusions The sensitivity of MRI in predicting SVI is poor,the specificity and accuracy are common.All the predictive tools have a reasonable value for SVI (AUC ＞ 0.7),and Gallina nomogram is superior to two versions of Partin tables in predicting SVI.

Objective To evaluate effectiveness of the applying pelvic double-slice retractor in the division of bladder and prostate pedicles in male patients with previous surgery history during radical cystectomy (RC) and anterograde radical prostatectomy (ARP). Methods The pelvic double-slice retractor was used to expose the bladder and prostate pedicles in 43 RC (30 cases with BMI ＞ 28 kg/m2,16 cases had previous partial cystectomy) and 37 ARP ( all cases with BMI ＞ 28 kg/m2 ). Results The pelvic double-slice retractor provided excellent exposure for the division of bladder and prostate pedicles.The handling of bladder and prostate pedicles became easy and safe without unnecessary bleeding and injury to the rectum.The median operating time to control the pedicles during RC and ARP were 12 min and 7 min,and the average blood loss were 30 ml and 20 ml,respectively. Conclusion The use of pelvic double-slice retractor for the exposure of the bladder and prostate pedicles is simple and effective in male patients with complicated pelvic anatomy during RC and ARP.

Objective To explore the alterations of protein phosphatase-2A (PP-2A) in lymphocytes in mild cognition impairment (MCI) and Alzheimer's disease (AD).Methods The activity PP-2A of was measured by ~(32)p liquid seintillography for incorporated radioactivity in control group(n=11) , the MCI group(n=11),and the AD group(n=11).The expression of PP-2A was determined by Western blot.Results In the control group,the activity of PP-2A (1.01?0.09) and the expression of PP-2A (0.96?0.07) were high while in the MCI group,the activity of PP-2A (0.71?0.12) and the expression of PP-2A (0.80?0.05) were decreased (both P