Objective To develop and verify a duplex real-time PCR method for simultaneous detection of bovine viral diarrhea virus(BVDV) and African swine fever virus(ASFV) in pig vaccines and apply it preliminarily.Methods According to the sequence of BVDV 5'UTR and ASFV VP72 included in NCBI,a pair of specific primers and a TaqMan probe were designed respectively,and a duplex real-time PCR method for simultaneous detection of BVDV and ASFV was established. The developed method were verified for the sensitivity,specificity and precision,and used to detect BVDV and ASFV of a total of 76 batches of live vaccines against porcine pseudorabies virus(PRV)and classical swine fever virus(CSFV),as well as live vaccines against porcine PRV added with BVDV and recombinant plasmid pMD-VP72,5 bottles for each,respectively. The BVDV and ASFV were rechecked and detected using the methods stipulated in the third part of Veterinary Pharmacopoeia of the People's Republic of China(2020 edition)and Announcement No. 172 of the Ministry of Agriculture and Rural Affairs of the People's Republic of China.Results 4. 2 copies/μL of BVDV and 8. 7 copies/μL of ASFV recombinant plasmids were detected by the developed duplex real-time PCR;No cross reaction with other common pathogens was observed and the coefficient of variation(CV)of repeatability and precision was not more than 2%.Conclusion The duplex real-time PCR method developed in this experiment showed good sensitivity,specificity and precision,which might be used for the rapid detection of exogenous viruses in pig vaccines and the quality control of pig vaccines and other biological products.

Objective To investigate protection by immunization with recombinant Ferritin vaccine of Echinococcus granulosus against protoscolices.Methods ICR mice were randomized into 3groups of 12 mice in each.The mice in group A and B were immunized three times with an interval of two weeks and those in group C did nothing.The animals in all the 3 groups were challenged with 1100 protoscolices intraperitoneally on the 8th week.Serum samples were collected before each inoculation and challenge injection.Seven months later, all the mice were killed and examinated for hydatid cysts.Result The number of cysts was significantly lower in the group A than in group B and C (P＜0.05).The levels of protection afforded were found to be 73% and 85%, respectively.Meanwhile,the number of cysts was markedly lower in group B than in group C(P＜0.05).The rate of protection afforded was 42%.Conclusion Recombinant Ferritin vaccine of Echinococcus granulosus shows partial immune protection.Therefore, it might be a suitable candidate for cocktail vaccine study in the future.

The present paper is aimed to detect superparamagnetic iron oxide labeled c-erbB2 oncogene antisense oligonucleotide probe (magnetic antisense probe) connected with SK-Br-3 oncocyte mRNA nucleotide by high resolution atomic force microscope (AFM). We transfected SK-Br-3 oncocyte with magnetic antisense probe, then observed the cells by AFM with high resolution and detected protein expression and magnetic resonance imagine (MRI). The high resolution AFM clearly showed the connection of the oligonucleotide remote end of magnetic antisense probe with the mRNA nucleotide of oncocyte. The expression of e-erbB2 protein in SK-Br3 cells were highly inhibited by using magnetic antisense probe. We then obtained the lowest signal to noise ratio (SNR) of SK-Br-3 oncocyte transfected with magnetic antisense probe by MRI (P<0.05). These experiments demonstrated that the high resolution AFM could be used to show the binding of magnetic antisense probe and SK-Br-3 mRNA of tumor cell nuclear.
Breast Neoplasms ; metabolism ; pathology ; Cell Line, Tumor ; DNA, Antisense ; chemistry ; genetics ; Female ; Ferric Compounds ; chemistry ; Genes, erbB-2 ; genetics ; Humans ; Magnetics ; Microscopy, Atomic Force ; methods ; Molecular Probe Techniques ; Nucleic Acid Probes ; chemistry ; genetics ; Oligodeoxyribonucleotides ; chemistry ; genetics ; Oxyphil Cells ; ultrastructure ; RNA, Messenger ; genetics ; metabolism

Objectives To explore the genetic diagnosis of fructose 1,6 diphosphatase deficiency and analysis of mutation sites of its pathogenic genes. Methods The clinical data and the related results of gene panel screening in one child with fructose 1, 6 diphosphatase (FBPase) deficiency were retrospectively reviewed. Results The 2-year-old girl suffered repeated infection, nausea, vomiting, mental illness, and drowsiness, accompanied by intermittent convulsions. Blood biochemical tests sμggested hypoglycemia and acidosis.The FBP1 gene had a missense mutation,c.355G>A,p.Asp119Asn(isozygoty).Both her parents carried the locus variation (heterozygous). Conclusions Fructose 1, 6 diphosphatase deficiency should be considered when child with hypoglycemia after repeated infection, acidosis, and ketosis.

OBJECTIVE: To explore the genetic basis for a fetus with autosomal recessive polycystic kidney disease (ARPKD). METHODS: Fetal tissue and peripheral blood samples were respectively obtained from the abortus and the couple. Following extraction of genomic DNA, genetic testing was carried out. RESULTS: The fetus was found to carry compound heterozygous variants of the PKHD1 gene, namely c.5336A>T (p.N1779I) and c.9455delA (p.N3152Tfs*10), which were respectively inherited from the husband and wife. CONCLUSION The c.5336A>T and c.9455delA variants of the PKHD1 gene probably account for the ARPKD in the fetus. Above results have enabled genetic counseling and prenatal diagnosis for the couple.