Establishment of A549 cell line with stable expression of HIF1α mediated by lentiviral vector / 重庆医学

Objective To establish the A549 cell line with stable expression of HIF1α by using lentiviral vector system.Methods Primers were designed and synthesized with human HIF1α gene coding sequence by the National Center of Biotechnogical Information(NCBI) as the template.HIF1α was amplified by PCR.The HIF1α fragment recycled by enzyme digestion was recombined with prepared lentiviral vector HBLV-RFP-Puro.The recombinant plasmid was identified by PCR and gene sequencing.The recombinant plasmid and the auxiliary plasmid were co-transfect into 293T cell.After filtration and concentration of packaged virus,the viral titer was detected by using the dilution counting method.The prepared lentivirus was infected A549 cells.The drug screening was adopted to stabilize the transfected cell line.The transfection effect was detected and observed by fluorescence microscope and Western blotting.Results The HIF1α fragment amplified by PCR was successfully verified and the recombinant plasmid was successfully constructed by PCR and gene sequencing identification.High-titer LV-HIF1α was obtained by successful package.After LV-HIF1α infecting A549 cells,the cells showed the red fluorescence by fluorescence microscope.The expression level of HIF1α in the LV-HIF1α group was significant higher than that in the control group by Western blot.Conclusion The 549 cell line with HIF1α stable expression mediated by lentivirus is constructed successfully.

Clinical application of MR diffusion-weighted imaging in evaluating the short-term curative effect of cryoablation for pancreatic carcinoma / 介入放射学杂志

Objective To discuss the clinical value of magnetic resonance diffusion-weighted imaging (MR-DWI) in distinguishing tumor remnants from tumor necrosis of pancreatic carcinoma after cryoablation treatment.Methods Conventional MRI T1WI,T2WI scan,DWI sequence and dynamic enhanced MRI scan were performed in 26 patients with pancreatic carcinoma who were received cryoablation treatment.The changes in MRI signals after cryoablation treatment were recorded.The apparent diffusion coefficient (ADC) values of the normal pancreas,preoperative tumor tissue,postoperative remnants and necrosis tissue were calculated,and the results were compared.The correlation between the ADC values and the size of the tumor was evaluated,and the differences in ADC values among the tumors that had different diameter,location and staging were statistically analyzed.Results Of the 26 patients,complete necrosis of tumor was obtained in 16.The necrotic tumor tissue displayed low-signal on T1WI,high-signal on T2WI and low-signal on DWI,with no enhancement on dynamic enhanced imaging.Active residual tumor tissue was detected in 9 patients,among them the residual tumor diameter ＞5 cm was seen in 7 patients;the residual rate was 34.6％.ADC values of the following tissue,from low to high in order,were preoperative pancreatic tumor tissue (1.022± 0.126)x10-3 mm2/s,postoperative residual tumor tissue (1.130±0.155)x10-3 mm2/s,normal pancreatic tissue (1.924±-0.124)×10-3 mm2/s and postoperative necrosis tissue (2.312-±0.214)×10-3 mm2/s.No statistically significant difference in ADC values existed between preoperative pancreatic tumor tissue and postoperative residual tumor tissue (P=0.452),while statistically significant difference in ADC values existed between normal pancreatic tissue and postoperative necrosis tissue (P＜0.001).The ADC values of pancreatic tumor tissue bore a negative correlation with the tumor size (R=-0.43,P=0.027 2),while the ADC values lacked the relationship to the tumor location as well as to the tumor staging (P=0.738 8 and P=0.089 5 respectively).Conclusion MR-DWI can effectively distinguish the residual tumor tissue from the necrotic tumor tissue of pancreatic carcinoma after cryoablation treatment,which provides reliable basis for further clinical diagnosis and treatment.

Expression of neuropeptide Y in rat testes and its significance / 中华男科学杂志

<p><b>OBJECTIVE</b>To assay the expression of neuropeptide Y (NPY) in rat testes and to investigate the significance of NPY in the regulation of androgen production and spermatogenesis.</p><p><b>METHODS</b>NPY mRNA levels in SD rat testes were measured by RT-PCR semi-quantity with beta-actin as internal control. NPY distribution was observed immunohistochemically.</p><p><b>RESULTS</b>NPY gene expressed in the testes, showing the strong positive band in the PCR production electrophoresis gel. In the immunostaining slides, NPY was found positively expressed in the Leyding cell area and around the testicular vessels and tubules, but not in the seminiferous tubules.</p><p><b>CONCLUSION</b>There was positive expression of NPY in the rat testes, which showed that NPY played a direct role in the regulation of testicular function.</p>

Effects of reproductive hormones on spermatogenesis / 中华男科学杂志

Spermatogenesis, a continuous course of cell proliferation and differentiation, depends on reproductive hormones. FSH and LH-induced T are the main hormones regulating spermatogensis. Intratesticular T is one of the key factors in maintaining spermatogenesis, while FSH is just as important for origination and maintenance of normal spermatogenesis. Sertoli cells are the pivot of hormonal regulation. Interestingly, these reproductive hormones also regulate sperm cell apoptosis spermatogenesis. Further studies on the hormonal regulation of spermatogenesis provide a base for the development of safe and recoverable contraceptives for males.