Effect of testosterone on pathological aggression behavior and expression of postsynaptics density-95 and growth associated protein-43 in prefrontal cortex in puberty rats / 第二军医大学学报

Objective To explore the effect of serum testosterone level on the pathologically aggressive behavior and the synaptic plasticity in the prefrontal cortex of puberty rats after gonadectomy. Methods Thirty male rats, 21 days old, were randomly divided into 3 groups: gonadectomized group, model group and control group. The gonadectomized and model groups were given a series of standard stress from 21 days old to puberty to induce animal model of pathological aggressive behavior. Resident-intruder experiment was performed to observe the variation of aggressive behaviors of animals. Enzyme-linked immunosorbent assay was used to examine the serum testosterone level. Western blotting analysis was used to determine the expression of postsynaptics density-95 (PSD-95) and growth associated protein-43 (GAP-43) in the prefrontal cortex. Results Resident-intruder experiment showed that the latency to the first attack in the gonadectomized group decreased significantly (P<0.01,P<0.01) and the attack times after yield increased significantly compared with those in the other two groups (P<0.01,P<0.01). The serum testosterone in the gonadectomized group was significantly decreased compared with the other two groups as shown by ELISA results (P<0.01, P<0.01). In addition, the aggressive-related behavior indices had a moderate to high negative correlation with the testosterone level (P<0.01). Western blotting analysis showed that prefrontal cortex expression of PSD-95 and GAP-43 in the gonadectomized group was significantly lower than those in the other two groups(P<0.01, P<0.01). Conclusion Low serum testosterone level can cause damage to the neuroplasticity of prefrontal cortex in puberty rats, which might be related to the pathologically aggressive behavior.

Expression of green fluorescent protein using an infectious cDNA clone of infectious bronchitis virus / 病毒学报

An infectious cDNA clone of H120 vaccine strain of infectious bronchitis virus (IBV) was constructed to demonstrate its potential as a gene transfer vector. Primers were designed according to the published genome sequence of H120 strain, and ten cDNA fragments covering the entire genome of H120 strain was amplified by RT-PCR. All the PCR products were ligated into pMD19-T vector and sequenced, and the ORF5a open reading frame in the pMDTM9 plasmid was replaced by an enhanced green fluorescent protein (EGFP) gene. Recombinant plasmids were digested by the restriction enzyme Bsa I, and all the cDNA fragments were recovered. By using appropriate ligation strategy, the genomic cDNA of H120 strain were reconstituted. Then genome RNA was synthesized in vitro by T7 RNA polymerase and transfected into BHK-21 cells. Recombinant virus expressing the green fluorescent protein was rescued and identified by RT-PCR and sequencing. The characteristics of recombinant virus were evaluated by passage in embryonated chicken eggs. This study showed that the 5a ORF is a good candidate for an insertion site of recombinant genes for the development of IBV infectious cDNA clone as a gene transfer vector.

Synthesis of 1-(1H-1, 2, 4-triazole-1-yl)-2-(2, 4-difluoro-phenyl)-3-(N-cyclopropyl-N-substituted amino)-2-propanols and its antifungal activity / 第二军医大学学报

Objective: To study the antifungal activity of triazole alcohols by introduction of cyclopropyl as side chain. Methods: Ten title compounds were synthesized and characterized by 1 HNMR,MS spectra and element analysis. The MICs of the compounds were determined by in vitro test in 8 fungus strains. Results: The title compounds exhibited potent antifungal activities against 8 strains,especially for the deep infection ones. Some compounds had MIC80 values less than 0.125 μg · ml-1 against Candida albicans, showing an activity 4 time higher than that of fluconzole. Conclusion: The title compounds with cyclopropyl and alky substituents have antifungal activities,and the antifungal activity decreases as the alkyl side chains getting longer.

Construction of a retroviral vector carrying HBX gene and its expression in LO2 human hepatocytes / 南方医科大学学报

<p><b>OBJECTIVE</b>To construct a retroviral vector carrying HBX gene and investigate its expression in LO2 human hepatocytes.</p><p><b>METHODS</b>HBX gene was amplified by PCR and subcloned into the retroviral vector pSEB-Flag to construct a retroviral plasmid (pSEB-Flag-HBX) expressing HBX. The HBX gene insert was confirmed by restriction enzyme digestion, PCR and DNA sequencing. The recombinant retroviruses carrying HBX gene were generated in 293T cells co-transfected with pSEB-Flag-HBX and the packaging plasmids pAmpho, and used to infect LO2 human hepatocyte. After selection with blasticidin, the mRNA and protein expressions of HBx were determined by the reverse transcription-PCR and Western blotting, respectively.</p><p><b>RESULTS</b>The retroviral plasmid (pSEB-Flag-HBX) carrying HBX was constructed successfully. The recombinant retrovirus efficiently delivered HBX gene into LO2 human hepatocyte, resulting in stable expression of HBX mRNA and HBx protein as shown by RT-PCR and Western blotting, respectively.</p><p><b>CONCLUSION</b>The recombinant retrovirus pSEB-Flag-HBX has been successfully constructed, which is capable of delivering the target gene HBX into LO2 human hepatocytes and results in stable expression of HBx to serve as an ideal model to study the effect of HBx on the development of hepatocellular carcinoma.</p>

Effect of vascular endothelial growth factor on pre-eclampsia in pregnant rats / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To investigate relationship between the vascular endothelial growth factor (VEGF) and the pathogenesis of pre-eclampsia in pregnant rats.</p><p><b>METHODS</b>Pregnant rats were divided into two groups randomly. Saline solution or L-nitro arginine methyl ester (L-NAME) 125 mg/d was given subcutaneously from day 7 of gestation till establishing pre-eclampsia. Systolic blood pressure, urine protein, platelet count, and weight of pups and placenta were determined. The levels of VEGF in pregnant rats venous serum, placenta and decidual tissue from normal pregnancy and pre-eclampsia rats were detected by ELISA and immunohistochemistry, respectively.</p><p><b>RESULT</b>Pregnant rats which were given L-NAME produced physical signs similar to those of pre-eclampsia, such as increase in systolic blood pressure [(145.3 +/-4.6)mmHg] and urine protein [(814.3 +/-57.5)mg/L], and decrease in platelet count [(467.1 +/-76.3) x 10(9)/L] and weight of pups and placenta. Compared with controls, the intensity of VEGF immunostaining in trophoblast or decidual cells were significantly reduced. The serum levels of VEGF were significantly lower in pre-eclampsia group than in normal pregnancy.</p><p><b>CONCLUSION</b>Decreased serum levels of VEGF and reduced expression of VEGF in placental tissues might in part explain the pathogenesis of pre-eclampsia in pregnant rats.</p>