Construction of goat germ cell specific reporting system pVASA-EGFP / 生物工程学报

To monitor the trans-differentiation from adult stem cells to germ cells, we analyzed the vasa expression of goat testicular tissues in different ages and constructed the germ cell specific reporting vector pVASA-EGFP. The expression of vasa was verified by RT-PCR and immunofluorescence. The vector pVASA-EGFP was constructed by molecular technology, then transfected into goat bone mesenchymal stem cells (BMSCs) by Lipofectamine 2000. Moreover, we observed the expression of the vector through green fluorescent protein (GFP). Immunofluorescence results show that Vasa was expressed in all groups of goat testicular tissues, RT-PCR results show that the levels of vasa mRNA in 3-month group and 10-month group were significantly higher than that in 10-day group. Sequencing and restriction enzyme results show that the vector was successfully constructed. After transfection and RA treatment, GFP expression was observed, which proved the validity of our reporting system. All the results proved that vasa was expressed in different ages in goat testicular tissues, and the vector pVASA-EGFP is efficient in monitoring the trans-differentiation in vitro, which paves the way for further characterization and screening of the trans-differentiation of goat BMSCs.

Effects of phycocyanin on apoptosis of human laryngeal cancer HEP-2 cells / 中国病理生理杂志

[ ABSTRACT] AIM:To investigate the effect of phycocyanin on the apoptosis of human laryngeal cancer HEP-2 cells and to explore the inhibitory mechanism of phycocyanin to tumor.METHODS:Highly purified phycocyanin was ex-tracted from spirulina.The effects of phycocyanin at different concentrations on the growth of human laryngeal cancer HEP-2 cells were detected by MTT assay.In addition, the cell structures were observed under electron microscope.The cell ap-optosis was analyzed by flow cytometry.The production of reactive oxygen species ( ROS) was measured by flow cytometry. Enzymatic activities of caspase-3,-8 and-9 were measured by chemical colorimatry.The expression of Bax, Bcl-2, Fas, P53, caspase-3 and caspase-9 at mRNA and protein levels was determined by RT-PCR and Western blot.RESULTS:MTT test confirmed that phycocyanin inhibited the cell activity of HEP-2 cells with time and dose dependent manners.The result of electron microscope observation and flow cytometry indicated that phycocyanin induced the apoptosis of HEP-2 cells.The intracellular content of ROS was increased.The activities of caspase-3, -8 and -9 were increased.RT-PCR showed that the mRNA expression of Bax, Fas, P53, caspase-3, caspase-9 was increased and Bcl-2 was decreased.The results of Western blot were consistent with the results of RT-PCR.CONCLUSION:Phycocyanin might induce apoptosis of HEP-2 cells by down-regulating Bcl-2, up-regulating Bax, Fas and P53, and the transduction of apoptotic signals in the human laryngeal cancer cells.

Study on correlation of androgen and androgen receptor with coronary heart disease in elderly men / 中华老年医学杂志

Objective To investigate the changes of sex hormone and androgen receptor levels and evaluate the relationship of the sex hormones and androgen receptor with coronary heart disease (CHD) in elderly men. Methods A cross-sectional study was performed in 539 elderly men, including 400 healthy people aged 62-92 years and 139 CHD patients aged 60-88 years. The plasma concentrations of total testosterone (TT), free testosterone (FT), dehydroepiandrosterone sulfate (DHEAS), sex hormone binding globulin (SHBG), estradiol (E2), luteinizing hormone (LH) and follicle-stimulating hormone (FSH) were measured. The androgen receptor (AR) level was tested by flow cytometry. Results The fluorescence intensity of DHEAS, TT, SHBG, FT and AR were significantly lower in CHD group than in healthy group (P<0.01);however, FSH and E2 in CHD group were higher(P(0.01). Age was negatively correlated with TT(r=-0.28,P<0.01) and FT (r=-0.17,P<0.05), and positively correlated with SHBG(r=0.14,P<0.05) and E2 (r=0.33, P<0.01). AR fluorescence intensity was negatively correlated with systolic blood pressure (r=-0.12,P<0.01). Logistic regression analysis indicated that TT (OR=1.065,9% CI: 1.012～1.121,P<0.05), SHBG(OR=0.994,95% CI:0.990～0.998,P<0.01) and AR (OR=0.971,95%CI:0.956～0.986, P<0.01)were significantly associated with CHD in elderly male patients. Conclusions The levels of DHEAS, TT, SHBG, FT and AR are lower in elderly men with CHD than in elderly healthy men;however, the FSH and E2 concentrations are higher. Low levels of TT, SHBG and AR may be the independent risk factors for CHD in elderly men.

Study of arousal in aged people with sleep apnea syndrome / 中华老年医学杂志

Objective To study the relationship between arousal and sleep apnea in old people with sleep apnea syndrome(SAS). Methods Sixty-two aged patients were diagnosed of SAS after polysomnography (PSG). Records included total sleep time, apnea-hypopnea index(AHI),total arousal number(TAN), arousal index(AI), total respiratory-related arousal number(TRAN), total number of arousal lasted over 15 seconds (T15AN) and the severity of hypoxemia (desaturation). Sixty-four non-aged patients were included in control group. Ten aged patients were intervened with nasal continuous positive airway pressure (nCPAP) treatment. Data were compared with that before treatment. Results The results for the above parameters of the aged and the non-aged SAS patients were( 35.1?23.2)/h and (39.1?27.2)/h respectively for AHI, (15.8?11.4)/h and (20.4?13.0)/h for AI, 44.1% and 51.3% for the ratio of TRAN/TAN, only 6.1% and 6.7% for ratio of T15AN/TAN. 50% of 62 aged and 64 non- aged SAS patients were in severe desaturation. The AI and TRAN of the aged patients were significantly lower than that of non-aged patients (P