AIM: To establish an HPCE method for determining the fingerprint of Compound Danshen Dropping Pill.(Radix et Rhizoma salviae miltiorrhizae,Radix et Rhizoma notoginseng,Borneolum Syntheticum) METHODS: A standard fingerprint was set up under following conditions: capillary,50 ?m (id)?50.0(eff.41.5) cm,uncoated;detector,UV 203 nm;injection,50 mbar(10 s);running voltage,20 kV;buffer,50 mmol/L sodium tetraborate and 50 mmol/L boric acid(pH 9.0,containing 30 mmol/L SDS) and acetonitrile(4∶1,v/v). RESULTS: Based on the fingerprints of 10 batches Compound Danshen Dripping Pills,an average electropherogram was used as the standard fingerprint,among which there were 14 characteristic peaks. CONCLUSION: The fingerprint analysis can be used for assessing the quality consistency of Compound Danshen Dropping Pill from batch to batch.

Objective To study the influence of inhibited steroidogenic factor-1 on human adrenocortical H295R cells, and explore its role in the pathogenesis of adrenal tumors. Methods The plasmids pGenesil1-SF-1-shRNA which containing U6 promoter and SF-1-specific short hairpin RNA (shRNA) and pGenesil1-negative-shRNA containing unspecific shRNA were transfected into H295R cell. The expression of SF-1 was measured by Western blot and real-time polymerase chain reaction(RT-PCR). Cell proliferation was analyzed by WST-1 assay and cell count. Ki-67 expression was detected by immunohistochemistry and cell apoptosis was examined by TUNEL assay. Results Compared with those in control cells, the protein and mRNA level of SF-1- transfected cells were reduced by 69.7% and 71.2% (P＜0. 01). WST-1 and cell count method showed that SF-1 gene silencing obviously inhibited cell proliferation(P＜0. 01). By contrast, there was a 3. 7-fold increase in the percentage of apoptotic H295R cells in SF-1-inhibited group than that of control group (P＜0. 01). Immunohistochemistry showed that Ki-67 positive cells in SF-1-inhibited cells were lower than the negative control cells (16.90±2.17) % and (33. 48±3.16)%,(P＜0. 01). Conclusion SF-1 gene silencing can inhibit the proliferation of adrenocortical cells, and it is expected to become a key protein in understanding pathogenesis of adrenal tumors or treating them.

Objective To investigate the relationship between raf kinase inhibitor protein (SKIP), a novel metastasis suppressor gene, and metastasis of ovarian carcinoma. Methods Immunohistochemistry, RT-PCR, and western blot analysis were performed to examine the expression of SKIP in clinical samples of ovarian tumors and five human ovarian carcinoma cell lines. Stable cell lines over-expressed or deleted of SKIP were cloned to investigate the function of SKIP in ovarian cancer cells. The recombinant plasmids expressing sense (ss) or antisense (as) SKIP cDNA or empty vector was transfected into ovarian cancer cell line SKOV3 by lipofectamine. The expression level of mitogen-activated protein kinase/extracellular signal-regulated kinase kinase (MEK) and extracellular signal-regulated kinase (ERK) in ovarian cancer cells were detected by western blot analysis. Assays of cell proliferation, soft-agar colony formation, cell adhesion, and cell invasion in vitro were used to examine the malignant phenotypes of the transfected cells. Flow cytometric analysis was performed to observe the effect of SKIP on cell cycle distribution before and after transfection. Results (1 ) The expression levels of SKIP protein in ovarian carcinoma tissues from patients were found to be reduced than those in ovarian benign tumor and borderline tumor. SKOV3 clones stably expressing full-length recombinant ssRKIP, asRKIP, and their respective empty vector were obtained. (2)RKIP was able to block basal levels of MEK and ERK in ovarian cancer cells. The expression level of phosphorylation MEK in ssRKIP#1 and ssRKIP#4 cells were 0. 35, 0. 34; while the expression level of phosphorylation ERK in ssRKIP#1 and ssRKIP#4 cells were 0.48 and 0.46. (3) Abilities of cell proliferation in the ssRKIP vector-transfected cells were decreased compared with that in the non-transfected cells (P ＜0. 01 ). (4)Anchorage-independent growth in the ssRKIP#1 and ssRKIP#4 cells (83.7 ± 5.7, 106. 0±9. 2) were decreased compared with that in the empty vector-transfected cells (158.3 ± 14. 6, P＜ 0. 01). (5)Cell adhesion in the ssRKIP#1 and ssRKIP#4 cells [(68.3±0. 8)%, (64. 1±0. 9)%] were decreased compared with that in the non-transfected cells [(100. 0 ± 1.1 )%, P ＜ 0. 01]. (6) Cell invasion in the ssRKIP#1 and ssRKIP#4 cells (24 ± 5, 25±4) were decreased compared with that in the non-transfected cells (68 ± 5, P ＜ 0. 01 ). (7) ssRKIP cells had a significant increase in the G1 phase and decrease in the G2 + S phase. Conclusion RKIP could inhibits the metastasis, but also the growth of ovarian cancer cells.

Objective:To identify the risk factors of anxiety disorders among patients undergoing coronary angiography (CAG) and to determine whether the decision of revascularization affect anxiety level following coronary angiography.
Methods:A total of 379 patients undergoing CAG in Fuwai Hospital from Dec. 2012 to Dec. 2013 were invited to participate this study. A data-collecting form, which included questions about demographic features, health history, Type A Behavior Questionnaire (TABQ) and Self Rating Anxiety Scale (SAS), was completed by the participants on the day before and the day after CAG.
Result:Among these patients, SAS score of both before and the day after CAG were higher than Chinese normative SAS score. Female patients had a higher SAS score level than male patients (40.57±9.53 vs 38.26±9.61, P<0.05) before CAG. Patients with these factors of female, over 50 years old, duration of coronary artery disease over 1 year, lower education level had a higher SAS score after CAG. SAS score declined signiifcantly after CAG except those scheduled to receive CABG. Multivariable linear analysis found the D-value between SAS scores before and after CAG was negative correlated with degree of education and positive correlated with the duration of coronary artery disease.
Conclusion:The anxiety level decreased after CAG, except those who need coronary revascularization surgery.

Objective To observe the effect of the optimized formulas of extracts of Radix Astragali and Radix Angelicae Sinensis on the survival status of the idiopathic pulmonary fibrosis (IPF) mice,and on the expression levels of transforming growth factor-β (TGF-β) and vascular endothelial growth factor(VEGF),so as to optimize the therapeutic regimen and to explore the therapeutic mechanism.Methods One hundred and five SPF ICR male mice were randomly divided into normal group,model group and 5 Chinese medicine treatment groups (group 1,2,3,4,5 of the optimized formula of Radix Astragali and Radix Angelicae Sinensis extracts).The mice in the model group and the 5 treatment groups were intratracheally injected with bleomycin (5 mg/kg) to induce the pulmonary fibrosis model.On day 21,the lung tissues were taken out for the test.Hydroxyproline content was detected by alkaline hydrolysis method,and morphological changes of lung tissues were observed by hematoxylineosin (HE) staining and Mallory's staining methods.The expression levels of TGF-β and VEGF were detected by polymerase chain reaction (PCR).Results The HE staining and Mallory's staining results showed that the pulmonary fibrosis in the 5 treatment groups was relieved as compared with that in the model group,especially in the group 1,and the alveolar structure recovered better.The 21-day overall death rate in the treatment groups were lower than those in the model group,and group 1 and group 5 had the lowest rates,the difference being statistically significant (P＜ 0.05).Compared with the model group,the content of hydroxyproline in the lung tissues of the treatment groups were decreased to some degrees,and there was significant difference (P ＜ 0.05 or P ＜ 0.01).The expression levels of TGF-β and VEGF in model group were higher than those in normal group,but were deceased in the treatment groups to some degrees,except TGF-β expression in group 5,and the difference was significant(P ＜ 0.05 or P ＜ 0.01).Conclusion When the contents of Radix Astragali water-extract and Radix Angelicae Sinensis alcohol-extract were predominated,the extract formula exerts certain effects on decreasing hydroxyproline content in the lung tissues,inhibiting the expression levels of TGF-β and VEGF,and relieving the degree of pulmonary fibrosis in IPF mice.

Objective To evaluate the key technique and effectiveness of potency preservation in robotic-assisted laparoscopic radical prostatectomy (RALP).Methods The complete clinical and follow-up data of 30 cases underwent RALP between February and May of 2016 were reviewed retrospectively.The average age of the patients was 67.3 years (48-82 years).The peak PSA level before surgery ranged from 7.6 to 53.4 ng/ml with the average level of 21.1 ng/ml.There were 7,16,6 and 1 case with the Gleason score of 6,7,8,and 9 point,respectively.Preoperative erectile score (IIEF-5) of the 30 patients were list as below:3 cases (0-10 points),4 cases(11-15 points),17 cases(16-20 points),and 6 cases(21-25 points).The key techniques of potency preservation during RALRP includes deep dorsal vein ligation techniques,neurovascular bundles preservation techniques and drain tube placement techniques.Results All 30 cases underwent operation successfully with no transfer to open surgery.Average operative time was 150.7 min (98-240 min) with an estimated blood loss of 165.7ml (50-550 ml).The average drainage removal time was 5.1 d postoperatively.The average bowel recovery time was 2.7 d and average hospitalization time was 8.2 d,respectively.There were two cases with one positive margin on the bladder neck and one on the tip of prostate,respectively.Seventeen cases gained potency preservation six months after surgery.Conclusion It was safe and beneficial for the potency preservation in RALP,which could be considered as one of the best options for the prostate cancer surgery.

Objective To investigate the in vitro effects of bladder cancer cell proliferation after silencing Notch1 gene. Methods The siRNA eukaryotic expression vector of Notch1 (psiRNA1)was constructed and transfected into bladder cancer cell lines T24 and BIU-87. Methabensthiazuron (MTT) and flow cytometry (FCM) assays were used to detect bladder cancer cells line growth, cell cycle and apoptosis after the transfection. RT-PCR and Western blotting were used to determine the expression changes of Notch1 in these cell lines. Results After transfection for 72 h, the rate of G0/G1 phase cells inceased from (23.89±1.32) % to (80.13±2.69)% in T24 cell line, and increased from (24.63±1.68)% to (69.44±2.41)% in BIU-87 cell line (both P<0.05). In addition, apop-totic cell index in T24 and BIU-87 cell lines increased from (1.28±0.14)% to (13.75±1.23)%, from (1.01±0.27)% to (8.72±1.01)%, respectively(both P<0.05). The growth of T24 and BIU-87 cell lines was obviously inhibited 24 h after the transfection, and the inhibitory effects lasted until 96 h after the transfection. Notch1 mRNA and protein significantly downregulated after transfection compared to the control(P<0.05). Conclusions Silencing Notch1 expression can inhibit the prolif-eration of bladder cancer cell lines. Notch1 gene might act as a tumor gene in bladder cancer.

The expression of angiotensin II type 1 receptor (AT(1)R) and angiotensin II type 2 receptor (AT(2)R) in aldosterone-producing adenoma (APA) of the adrenal gland was detected, and their relationship with clinical indexes of APA was analyzed. The mRNA expression of AT(1)R and AT(2)R in 50 cases of APA and tissues adjacent to tumors and 12 cases of normal adrenal tissues was detected by using reverse transcriptase polymerase chain reaction (RT-PCR). The expression of AT(1)R and AT(2)R proteins in paraffin-embedded slices of tissue was detected by immunohistochemistry. The expression of AT(1)R in adenoma, tissues adjacent to tumor, and normal tissues of the adrenal gland showed no significant differences. The expression of AT(2)R in APA tissue was lower than that in normal adrenal gland tissues (P<0.05). Correlation analysis of the mRNA expression level of AT(2)R and clinical data from patients demonstrated that AT(2)R expression was negatively related to plasma aldosterone concentration (PAC) (r=-0.467, P<0.05), but positively related with plasma renin activity (PRA) (r=0.604, P<0.05). It is concluded that down-regulation of the AT(2)R expression is possibly related with the tumorigenesis of APA.

This study examined the association of polymorphisms in angiotensin II receptor genes (AT (1) R and AT (2) R) with the risk for aldosterone-producing adenoma (APA) in a Chinese Han population. Four polymorphisms including rs5182 (573T/C) in exon 4, rs5186 (1166A/C) in 3'-untranslated region (3'-UTR) in AT (1) R gene and rs5194 (2274G/A) in 3'-UTR, rs1403543 (1675G/A) in intron 1 in AT (2) R gene were detected in 148 APA patients and 192 normal subjects (serving as control) by using a MGB-Taqman probe. The distribution of genotypes of each locus was in accordance with Hardy-Weinberg Equilibrium (HWE) in the APA and control groups (P>0.05). The allele A frequency at rs5194 was significantly higher in the APA group (0.49) than in the control group (0.35) (χ (2)=12.08, P=0.001). Subjects with homozygotic genotype AA and heterozygotic genotype GA were at an increased risk for APA as compared to those with GG genotype (OR=2.66, 95% CI=1.45-4.87; OR=1.67, 95% CI=1.02-2.74). Furthermore, rs5194 single-nucleotide polymorphism (SNP) at AT (2) R gene was significantly associated with APA in additive (OR=1.64, 95% CI=1.21-2.20, P=0.001), dominant (OR=1.94, 95% CI=1.23-3.06, P=0.003), and recessive model (OR=2.01, 95% CI=1.17-3.45, P=0.01). It was concluded that rs5194 polymorphism at AT (2) R gene was associated with the risk for APA, which may constitute a genetic marker of APA.

MicroRNAs (miRNAs or miRs) are a class of short, non-coding RNAs that participate in various oncological processes. This study aims to explore the roles of microRNA-34a (miR-34a) in invasive urothelial bladder carcinoma. miR-34a was transfected into bladder cancer cell lines 253J and J82. The miR-34a expression levels in tissues and cells were detected by using qRT-PCR. The Notch1 expression was detected by qRT-PCR and Western blotting. Cell migratory and invasive abilities were measured by Transwell chamber assay. Bioinformatics and luciferase assay were performed to predict and analyze the binding sites between miRNA-34a and Notch1. It was found that there was aberrant expression of miR-34a in bladder cancer tissues. Moreover, we revealed that ectopic expression of miR-34a suppressed cell migration and invasion, while forced expression of Notch1 increased cell migratory and invasive abilities. Finally, we observed that miR-34a transfection significantly down-regulated luciferase activity and reduced the mRNA and protein levels of Notch1. Our study concluded that microRNA-34a antagonizes Notch1 and inhibits cell migration and invasion of bladder cancer cells, which indicates the tumor-suppressive function of microRNA-34a in bladder cancer.