Construction of lentiviral mediated CyPA siRNA and its functions in non-small cell lung cancer / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To construct a lentiviral-vector-mediated CyPA small interference RNA (siRNA) and study its function in non-small cell lung cancer.</p><p><b>METHODS</b>First, four target sequences were selected according to CyPA mRNA sequence, the complementary DNA contained both sense and antisense oligonucleotides were designed, synthesized and cloned into the pGCL-GFP vector, which contained U6 promoter and green fluorescent protein (GFP). The resulting lentiviral vector containing CyPA shRNA was named Lv-shCyPA, and it was confirmed by PCR and sequencing. Next, it was cotransfected by Lipofectamine 2000 along with pHelper1.0 and pHelper 2.0 into 293T cells to package lentivirus particles. At the same time, the packed virus infected non-small cell lung cancer cell (A549), the level of CyPA protein at 5 d after infection was detected by Western Blot to screen the target of CyPA. A549 were infected with Lv-shCyPA and grown as xenografts in severe combined immunodeficient mice. Cell cycle and apoptosis were measured by FCM.</p><p><b>RESULTS</b>It was confirmed by PCR and DNA sequencing that lentiviral-vector-mediated CyPA siRNA (Lv-shCyPA) producing CyPA shRNA was constructed successfully. The titer of concentrated virus were 1 x 10(7) TU/ml. Flow cytometric analysis demonstrated G2-M phase (11.40% +/- 0.68%) was decreased relatively in A549/LvshCyPA compared with control groups (14.52% +/- 1.19%) (P<0.05). The apoptosis rate of A549/Lv-shCyPA (5.01% +/- 0.5%) was higher than control groups (0.35% +/- 0.17%) (P<0.05). Visible tumors were only detectable at 6th day after inoculated by A549/Lv-shCyPA. The xenograft tumors of A549/Lv-shCyPA remarkably delayed tumor growth and remained at a similarly small average size at 38th days after inoculation compared with the control group (P < 0.05).</p><p><b>CONCLUSION</b>Lentiviral-vector-mediated siRNA technique effectively inhibits the expression of CyPA, induces the NSCLC cell apoptosis, inhibits the tumor growth. Elucidation of the precise role of CypA in these pathways may lead to new targeted therapies for non-small cell lung cancer.</p>

Antagonistic effects of vitamin E on the testicular injury by cyclophosphamide in mice / 中华男科学杂志

<p><b>OBJECTIVE</b>To observe the protective effects of vitamin E on the testicular injury by cyclophosphamide in mice, and the correlative mechanism.</p><p><b>METHODS</b>Fifty sexually mature male mice were randomly divided into five groups: the cyclophosphamide group (the CP group), the low-dose vitamin E group (the low-dose group), the middle-dose vitamin E group (the middle-dose group), the high-dose vitamin E group (the high-dose group), the matched control group (the control group). The first four groups were given cyclophosphamide by gavage at a dose of 5 mg/(kg x d). The low-dose group, the middle-dose group and the high-dose group were given vitamin E by subcutaneous injection at doses of 30 mg/(kg x d), 50 mg/(kg x d) , 70 mg/(kg x d) after 4 h of cyclophosphamide treatment. The control group was gavaged with equivalent normal saline. The treatment period for all groups was 28 days. The level of plasma FSH, LH, T and the activity of testicular SOD, GSHPx, CAT and the level of testicular MDA were detected. The histological structure and the ultrastructure of the testis were examined by light microscope and electron microscope.</p><p><b>RESULTS</b>As compared with the CP group, the plasma FSH, LH, T level and the SOD, GSHPx, CAT activity in the middle-dose group and the high-dose group were higher (P< 0.05, P< 0.01), MDA level significantly lower(P<0.01). The histological structure and the ultrastructure of the testis were in the normal range.</p><p><b>CONCLUSION</b>Vitamin E has protective effects on the testicular injury by cyclophosphamide in mice. The possible mechanism of vitamin E may be its scavenging free radical and antioxidant effects, as well as it may have some stimulatory effects on gonadotrophin releasing of pituitary anterior lobe.</p>

Analysis of sterile male semen of occupational drivers / 中华男科学杂志

<p><b>OBJECTIVE</b>To explore the possible correlation between the driver's occupation and male semen quality.</p><p><b>METHODS</b>Semen samples were collected from 1,223 infertile men (78 drivers and 1,145 non-drivers) and 100 normal men, and their liquefaction, sperm density, sperm vitality, sperm motility and sperm shape were analysed.</p><p><b>RESULTS</b>The abnormal rates of semen quality in sterile male drivers were significantly higher than in non-drivers(P < 0.05) and in normal men(P < 0.01). The semen abnormal rates in drivers with more than 8 years' driving experience were higher than in those with less than 8 years' driving experience(P < 0.05).</p><p><b>CONCLUSION</b>Driving occupation can result in abnormal semen quality.</p>