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1.
China Pharmacy ; (12): 2632-2634,2635, 2015.
Article in Chinese | WPRIM | ID: wpr-605137

ABSTRACT

OBJECTIVE:To establish a method for determining plasma concentrations of tanshinone ⅡA and use it for pharma-cokinetics study of tanshinone ⅡA in Beagle dogs. METHODS:HPLC was performed on the column of Phenomenex Luna C18, with the mobile phase of water(5 mmol/L of ammonium acetate+0.05% H3PO4)-acetonitrile at flow rate of 1.0 ml/min;the detec-tion wavelength was 270 nm,column temperature was 40 ℃ and the volume was 20 μl. 9 Beagle dogs were randomly divided into tanshinone ⅡA low,medium and high dose groups(2,4 and 8 mg/kg),blood was respectively taken from forelimb venous blood to determine the plasma concentration before and after 2,5,10,15,20,30,45,60,75,90 and 120 min of iv administration. DAS 2.0 software was used to calculate the pharmacokinetic parameters. RESULTS:The linear range of tanshinoneⅡA was 0.097 5-12.50 μg/ml (r=0.999 8);the RSDs of precision and stability tests were all less than 10%;the method recovery was 100.4%-107.2%,and extraction recovery was 90.2%-92.3%. The t1/2αin tanshinoneⅡA low,medium and high dose groups were respective-ly(1.01±0.27),(1.03±0.46)and(1.51±0.65)min;t1/2β were respectively(16.25±4.78),(22.42±9.32)and(24.45±12.02) min;AUC0-120 min were respectively(150.88±45.25),(305.44±92.55)and(643.67±178.27)μg·min/ml;and CL were respective-ly(12.01±4.36),(12.78±5.06)and(12.17±5.41)ml/(min·kg). CONCLUSIONS:The precision,stability and recovery of the method are all in line with the determination requirements of biological samples,and tanshinone ⅡA showed a good linear relation-ship with dose in Beagle dogs in AUC0-120 min.

2.
Chinese Journal of Biotechnology ; (12): 1138-1143, 2009.
Article in Chinese | WPRIM | ID: wpr-296946

ABSTRACT

In this study, we evaluated the development potential of caprine mammary gland epithelial cells (CMGECs) after transfection and nuclear transfer into enucleated, ovulated oocytes. We first isolated CMGECs from udders of lactating goats which were transfected with expression plasmid for human lacterrin and selected by G418. Then we chose sixteen neomycin resistant lines and induced them with prolactin for the expression of human lactoferrin checked by Western blotting. The donor cells, expressing human lactoferrin of 75 kD, were fused and activated with enucleated ovulated oocytes. Pronuclear-stage reconstructed embryos were transferred into the oviducts of 16 recipient goats. There were fourteen (87.5%), thirteen (81.3%), and ten (62.5%) pregnancies confirmed pregnant by ultrasound on Day 30, 60, and 90, respectively. Three recipients carried the pregnancies to term and delivered one goat each. Nested PCR-RFLP analysis confirmed that all of the kids were clones of the donor cells. These results demonstrated that CMGECs after transfection remain totipotent for nuclear transfer.


Subject(s)
Animals , Cloning, Organism , Methods , Epithelial Cells , Cell Biology , Female , Goats , Humans , Lactoferrin , Mammary Glands, Animal , Cell Biology , Nuclear Transfer Techniques , Pregnancy , Transfection
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