Construction and identification of recombination expression vector Ksp-Cadherin-Gpx1-Kik1 / 药物分析学报

Objective To construct and identify the Gpx1-Klk1 vector which contains kidney-specific promoter (Ksp-cadherin). Methods Through PCR amplification, the human Gpx1, Klk1, and Ksp-cadherin eDNA were obtained by taking Gpx1 cDNA, Klk1 eDNA, and Ksp-cadherin BAC as templates. After being testified, the PCR products were inserted into the expressive vector pIRES-EGFP step-by-step to produce a recombinant vector Ksp-cadherin-Gpx1-Klk1. This vector was examined by restriction enzyme digestion and sequence analysis. Results The recombinant expressive vector Ksp-cadherin-Gpx1-Klk1 was successfully constructed. Conclusion The construction of the recombinant vector Ksp-cadherin-Gpx1-Kik1 laid foundations for investigations in establishing transgenic animal models, the over-expression of Gpx1 and Kikl in mammal kidney, and gene therapy for ischemia-reperfnsion injury during kidney transplantation.

Construction of a Transgenic Mice Model with High Serum Titer of Natural Anti-keratin Autoantibody / 中华皮肤科杂志

Objective To construct an anti-keratin autoantibody (AK auto Ab) transgenic mouse model. Methods Linearized transgene plasmid was microinjected into the zygotes of CBA?C57BL/6 mice, which were transplanted into the oviducts of pseudo-pregnant mice. PCR was used to identify the genotype of the offsprings, and ELISA was applied to measure the serum levels of AK auto Ab. Results Twelve transgene positive founder mice were obtained, and 9 of them produced offsprings as the third generation. The serum level of AK auto Ab was increased in 3 of the transgenic mice. Conclusions AK auto Ab transgenic mice were successfully established; these mice could serve as an animal model with increased serum levels of AK auto Ab.

Alterations of cardiac sympathetic norepinephrine transporter expression in rats with volume overload / 中国病理生理杂志

AIM: To determine the alterations of myocardial ? 1-adrenergic receptor (? 1-AR) and cardiac sympathetic norepinephrine transporter (NET) mRNA expression, which is upstream modulator of ? 1-AR, in rats with longterm volume overload (VOL).METHODS: Left ventricular systolic (LVSP) and end diastolic pressure (LVEDP) of rats with VOL induced by aortacaval fistula operation and control group were measured at 3, 14,30 and 60 d after the operation, the mRNA at the time points was measured by RT-PCR and Northern blot analysis and quantified by densitometry.RESULTS: The cardiac sympathetic NET specific expression is in the cardiac sympathetic ganglia. Be compared with the control group, LVSP of VOL rats decreased most dramatically by 24%( P