OBJECTIVE:To provide reference for the improvement of supervision system of online drug sale in China. METH-ODS:Usingonline pharmacyonline drugstoreinternet drugssupervisionas keywords,CNKI,Wanfang database from Jan. 2015 to Jan. 2016 and related websites were retrieved,experience about legislation,legal provisions and regulatory bodies in the United States,Britain,Germany and other countries were summarized. The problems existing in the current supervision system in China were analyzed,and related suggestions were proposed. RESULTS & CONCLUSIONS:The legal system of the online drug sale in the United States,Britain,Germany and other countries is more complete,legislation content is relatively perfect,pay-ing attention to the combination of administrative supervision and industry supervision. The supervision system of online drug sale in China exists problems as follows as legislation level relatively lower,conflict between laws and regulations;the legislation con-tent being not perfect,contents missing or provisions blur in many aspects;law enforcement existing in the government depart-ments,industry regulation being weak and others. It is suggested that the country should develop high-order law when improving the supervision system of online drug sale,modifying the existing law or promulgating the relevant specialized laws by legal form rather than regulations and rules;develop clear,actionable law and learn from foreign related systems (such as electronic evi-dence,blacklist)to improve legal contents;and further clear the powers and responsibilities of the regulatory authorities,actively develop industry associations(such as pharmaceutical association,etc.)to conduct supervision for online drug sale.

To knock out β-lactoglobulin (BLG) gene and insert human lactoferrin (hLF) coding sequence at BLG locus of goat, the transcription activator-like effector nucleases (TALEN) mediated recombination was used to edit the BLG gene of goat fetal fibroblast, then as donor cells for somatic cell nuclear transfer. We designed a pair of specific plasmid TALEN-3-L/R for goat BLG exon III recognition sites, and BLC14-TK vector containing a negative selection gene HSV-TK, was used for the knock in of hLF gene. TALENs plasmids were transfected into the goat fetal fibroblast cells, and the cells were screened three days by 2 μg/mL puromycin. DNA cleavage activities of cells were verified by PCR amplification and DNA production sequencing. Then, targeting vector BLC14-TK and plasmids TALEN-3-L/R were co-transfected into goat fetal fibroblasts, both 700 μg/mL G418 and 2 μg/mL GCV were simultaneously used to screen G418-resistant cells. Detections of integration and recombination were implemented to obtain cells with hLF gene site-specific integration. We chose targeting cells as donor cells for somatic cell nuclear transfer. The mutagenicity of TALEN-3-L/R was between 25% and 30%. A total of 335 reconstructed embryos with 6 BLG-/hLF+ targeting cell lines were transferred into 16 recipient goats. There were 9 pregnancies confirmed by ultrasound on day 30 to 35 (pregnancy rate of 39.1%), and one of 50-day-old fetus with BLG-/hLF+ was achieved. These results provide the basis for hLF gene knock-in at BLG locus of goat and cultivating transgenic goat of low allergens and rich hLF in the milk.
Animals ; Animals, Genetically Modified ; genetics ; Female ; Fibroblasts ; Gene Knock-In Techniques ; Gene Knockout Techniques ; Goats ; genetics ; Humans ; Lactoferrin ; genetics ; Lactoglobulins ; genetics ; Milk ; chemistry ; Nuclear Transfer Techniques ; Plasmids ; Pregnancy ; Transfection