Objective To separate the side population cells(SP) from breast cancer MCF-7 cell line,and observe its biological characteristics.Methods Flow cytometry and Hcechst 33342 dye efflux assay were used to isolate SP cells and non-SP cells from the MCF-7 cell line of human breast cancer.Tumorigenicity of the two subpopulations was observed by a soft agar cloning method.Results The results of FACS analysis indicated that (6.5 ± 0.4 ) ％of the MCF-7 cells were SP cells;The vitro colony formation rate of SP cells was(38.5 ±9.4)％,and higher than that of non-SP cells ( 8.4 ± 2.6 ) ％ ( t =5.34,P ＜ 0,05 ).Concluslon The SP cells sorted from MCF-7 cell line enriched tunor stem cells,which exhibited high tumorigenicity.It indicated that SP cells should play a principal role in breast cancer.

Brain Neoplasms ; pathology ; Glioma ; pathology ; Humans ; Infant, Newborn ; Male

Female ; Humans ; Infant, Newborn ; Kidney Neoplasms ; Twins, Monozygotic ; Wilms Tumor

Animals ; Arrhythmias, Cardiac ; etiology ; therapy ; Captopril ; therapeutic use ; Disease Models, Animal ; Electric Stimulation Therapy ; methods ; Myocardial Ischemia ; physiopathology ; therapy ; Rabbits ; Ventricular Fibrillation

Animals ; Cochlea ; pathology ; Female ; Guinea Pigs ; Hyperlipidemias ; pathology ; physiopathology ; Male ; Otoacoustic Emissions, Spontaneous

Objective To evaluate the short-term efficacy of Perigee system in the treatment of anterior pelvic organ prolapse. Methods From October 2012 to September 2014, 59 patients with pelvic organ prolapse, pelvic organ prolapse quantitation system (POP-Q) were diagnosed as anterior pelvic organ prolapse Ⅲ degree and above were performed Perigee anterior pelvic floor reconstruction, while some patients combined with sacrospinous ligament suspension, posterior wall repair or posterior pelvic reconstruction surgery for pelvic prolapses. Pelvic floor distress inventory-short form 20 (PFDI-20), pelvic organ prolapse-urinary incontinence sexual questionnaire-12 (PISQ-12) were evaluated, and postoperative POP-Q were used to analyze the changes of the indexes and postoperative complications. Results In 59 patients, the average operation time was (99±29) minutes, the average intraoperative blood loss was (119± 92) ml. The median postoperative follow-up time of 59 cases was 17.5 months (range:8-30 months), median follow-up time of subjuctive symptoms was 21.2 months (range:11-34 months), the total score of PFDI-20 was compared with the preoperative, and the difference was statistically significant (5.6 versus 27.8, P<0.01). It was statistically significant of PISQ-12 score before and after surgery (34±3 versus 36±4, P<0.05). Short-term anatomical cure rate was 98%(58/59), 1 cases (2%, 1/59) in recurrence, 2 cases (3%, 2/59) of erosion. Conclusion This results show that the Perigee system is effective and reliable in the treatment of anterior pelvic organ prolapse.

Objective To investigate the optimal extraction technique and antioxidant activity of curcuminoids from Curcuma longa L. Methods The optimized extraction process of curcuminoids from Curcuma longa L. with ethanol as extracting solvent were studied based on orthogonal test.The effect of each factor such as the ethanol concentration, the amount of solvent, extraction temperature and extraction time were investigated based on a single-factor experiment.The effects of extraction yield of curcuminoids under different processing conditions were compared by HPLC method. Content of curcuminoids and antioxidant activity of the extract were used as comprehensive survey index. Free radical scavenging DPPH method was used to assess the antioxidant effect. And the extraction technique parameters were optimized. Results The condition of optimal extraction technique was: ethanol concentration 60%, amount of solvent 20 folds, extraction time 2 h, extraction temperature 90 ℃. Under these conditions, the extraction yield of curcuminoids was 3. 65%. DPPH free radical scavenging yield of the curcumin extract was 79.79%. Conclusion This optimized technology is simple and have a good reappearing rate, and the extracts of Curcuma longa L.show good antioxidant activity.

Objective To investigate the effects of 5-azacytidine on radiation-induced epithelialmesenchymal transition in rat alveolar type Ⅱ epithelial cell line (RLE-6TN) and explore their working mechanisms,and to provide experimental evidence for the potential drug-based treatment of radiationinduced pulmonary fibrosis.Methods RLE-6TN cells were cultured in vitro and divided into four groups according to the experimental purposes:control group (C),radiation group (R),5-azacytidine group (A),and radiation followed by 5-azacytidine group (R + A).The microstructural changes in cells were determined by transmission electron microscopy.Inverted phase-contrast microscopy revealed the morphological changes in cells.The mRNA expression levels of E-cadherin and α-SMA were measured by quantitative real-time polymerase chain reaction (qRT-PCR).The protein expression levels of E-cadherin,GSK3 β,and p-GSK3 β (Ser9) were measured by Western blot.The one-way analysis of variance was used for pairwise comparison.Results The cells in group R became spindle-like.Similar morphological changes were not observed in cells in group R + A.Osmiophilic lamellar bodies disappeared at last in cells in group R.RT-PCR results showed that compared with group C,group R had a significantly lower mRNA expression level of E-cadherin ((0.23 ± 0.06) vs.(1.00 ± 0.00),P =0.002)) and a significantly higher mRNA expression level of α-SMA ((2.91 ± 0.01) vs.(1.00 ± 0.00),P =0.000)).However,compared with group R,group R + A had a significantly higher mRNA expression level of E-cadherin ((0.47 ± 0.05) vs.(1.00 ± 0.00),P =0.024)) but a significantly lower mRNA expression level of α-SMA ((2.50 ± 0.02) vs.(1.00 ±0.00),P =0.037)).The results of Western blot showed that the protein expression level of Ecadherin was significantly reduced ((0.07 ± 0.01) vs.(0.48 ± 0.02),P =0.028)),while the protein expression level of p-GSK3β was significantly increased in Group R than in Group C ((0.85 ± 0.04) vs.(0.23 ± 0.03),P =0.031)).However,compared with group R,group R + A had a significantly lower protein expression level of E-cadherin ((0.25 ± 0.00) vs.(0.07 ± 0.01),P =0.024)) and significantly less up-regulation of the protein expression level of p-GSK3β ((0.39 ± 0.03) vs.(0.85 ± 0.04),P =0.014)).Conclusions X-ray radiation can induce the epithelial-mesenchymal transition in epithelial cells.5-azacytidine suppresses radiation-induced epithelial-mesenchymal transition by inhibition of the activity of p-GSK3β in RLE-6TN cells.

OBJECTIVE To establish a method for determination of Nimesulide in suppositories. METHODS Screening different organic solvent to extract 36 type semi-sythetic fatty acid ester in suppositories and determinating Nimesulide by UV. RESULTS N-Hexano was used as the solution of extraction. A good linearity was within the concentration range of 3～15μg/mL with a Borrelation coefficient of 0.9999. The average recovery was 98.3%. The RSD for reproducibility was 0.47%. CONCLUSION The method is simple, accurate and reliable.

Objective To explore the mechanism of the formation of glioma cells derived vessels (GCDV).Methods The tubular formation assay was performed on 3D cell cultures of U87,U251,U373,SF295,T98G,SKMG-4 and C6 glioma cell lines.The expression of Notch1 、Dll4 were examined by western blot analysis.Result The mean number of vasculogenic channels of cell lines C6,U373,U87,T98G,SF295,U251 and SKMG-4 per area (100×) were 25.2±5.0,36.4±3.2,19.0±2.2,12.6±2.4,4±2.2,0.2±0.4 and 0,respectively.The relative protein densities of Notchl in U87,U251,T98G,SF295,SKMG-4,C6 and U373 were 0.34,0.21,0.79,0.04,0.28,1.75 and 1.19,which were significantly related with the tubular formation ability (P＜0.05).However,the expression of Dll4 was not associated with tubular formation ability (P＞0.05).Conclusion Notch1 may play a key role in tubular formation of glioma cells while the role of Dll4 in the process needs further study.