Construction of a lentiviral vector carrying short?hairpin RNA targeting PAX6 and its effect on proliferation of glioma U251 cells in vitro / 南方医科大学学报

<p><b>OBJECTIVE</b>To construct a lentiviral vector for delivering short hairpin RNA (shRNA) targeting PAX6 and investigate its effect on the proliferation of glioma U251 cells in vitro.</p><p><b>METHODS</b>Two small interfering RNA sequences targeting PAX6 gene were designed based on the reported sequence of PAX6 and annealed to form a double?stranded chain, which was inserted into a lentiviral vector to construct the recombinant lentiviral vector shRNA?PAX6. The recombinant vector was infected into U251 cells, and the expression of PAX6 mRNA and protein in the cells was detected by real?time PCR and Western blotting, respectively. The changes in the proliferation of U251 cells after the infection was assessed using MTT assay.</p><p><b>RESULTS</b>Double enzyme digestion of the lentiviral vector pLKD?CMV?G&NR?U6?shRNA yielded an 8208?bp fragment, and colony PCR and sequencing analysis confirmed successful construction of the lentiviral vector shRNA?PAX6. Infection of the cells with shRNA?PAX6 caused a significant reduction of the expressions of PAX6 mRNA and protein (P<0.05) and resulted in obviously increased proliferation of U251 cells (P<0.05).</p><p><b>CONCLUSION</b>We successfully constructed the recombinant vector shRNA?PAX6 for silencing PAX6 gene. PAX6 gene silencing results in increased proliferation of U251 cells in vitro.</p>

Effects of hypoxic preconditioning on hypoxia tolerance of astrocytes in vitro / 中国应用生理学杂志

<p><b>AIM</b>To explore the mechanisms of hypoxic preconditioning on protecting cultured astrocytes from hypoxia injury.</p><p><b>METHODS</b>Cultured astrocytes were divided randomly into several groups: control(C), hypoxia(H) and hypoxic preconditioning (HP). Cells MTT metabolic activity, qualitation of apoptosis and modality to explore the protection effects of hypoxic preconditioning. Immunocytochemistry of Bcl-2 and Bax to explore the mechanisms of hypoxic preconditioning on protecting astrocytes from hypoxia.</p><p><b>RESULTS</b>Compared with H group there was marked increase of MTT metabolic activity in HP48 and HP72 groups. Immunocytochemistry of Bcl-2 and Bax showed that compared with H group, expression of Bcl-2 was increased in HP group, while expression of Bax was decreased in HP group.</p><p><b>CONCLUSION</b>Hypoxic preconditioning can protect astrocytes from hypoxia. One possible mechanism maybe concerned with inhibition of Bax and maintain of Bcl-2 to depress apoptosis procedure.</p>

Correlation between MHC class I-related chain A gene *008 allele and human cytomegalovirus infection / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the correlation between MHC class I-related chain A (MICA) gene *008 allele and human cytomegalovirus (HCMV) infection.</p><p><b>METHODS</b>MICA*008 allele was detected in 86 patients with chronic granulocytic leukemia and 81 unrelated normal individuals by way of sequence-specific primers (PCR-SSP). Anti-HCMV IgM was also detected in the sera of these subjects with enzyme linked immunosorbent assay (ELISA).</p><p><b>RESULTS</b>MICA*008 allele frequency was lower in patients with chronic granulocytic leukemia than in the control group (22.2% vs 34.3%, Chi(2)=4.98, P<0.05). The infection rate of HCMV was significantly higher in those individuals with genotype of MICA*008 (-) than in those with MICA*008 (+), and moderate correlation was suggested between MICA*008 and HCMV infection (C=0.5829, 0.6142).</p><p><b>CONCLUSION</b>Individuals with MICA*008 positivity is not liable to HCMV infection, but those with MICA*008 (-) can be vulnerable to HCMV infection, suggesting an inverse correlation between MICA*008 allele with HCMV.</p>

MICA*008/A5.1 allele and HCMV infection in kidney transplanted donees of Hunan Han nationality / 中南大学学报(医学版)

OBJECTIVE@#To investigate the relationship between MICA*008/A5.1 allele and human cytomegalovirus (HCMV) infection in kidney transplanted donees of Hunan Han nationality.@*METHODS@#The MICA*008/A5.1 allele based on 91 kidney transplanted donees and 81 unrelated normal individuals of Han nationality in Hunan Province were analyzed by PCR/SSP assay. At the same time, anti-HCMV antibody IgM was detected in the serum by ELISA method.@*RESULTS@#The positive rate of MICA*008/A5.1 allele was significantly higher in the control group (56.79%) than that in the kidney transplanted donee group (34.07%) (P <0.05). The infection rate of HCMV in those individuals whose genotype was MICA*008/A5.1 (-) was significantly higher than that in the MICA*008/A5.1(+).@*CONCLUSION@#The individual whose genotype is MICA*008/A5.1 (+) is not liable to HCMV infection, but the individual whose genotype is MICA*008/A5.1 (-) is liable to HCMV infection.