| Prion diseases are one of degenerative disorders of the nervous system. Prion diseases are also known as transmissible spongiform encephalopathies (TSEs) caused by infectious protein(Prion protein, PrPSc)which are convered from normal cellular proteins (PrPc) without nucleic acid. Human prion diseases are classified as sporadic Creutzfeldt-Jakob disease (CJD), vCJD, Gerstmann-Straussler-Scheinker disease, Kuru and Fatal familial insomnia. Protein synthesis of mRNA can bestarted by two mechanisms: cap-dependent translation initiation and cap-independent translation initiation. Cap-independent translation initiation is propelled by internal ribosome entry sites (IRESs) that are found in both viral RNAs and cellular mRNAs. To explore whether the translation initiation mechanism of the Prion protein, PrP eukaryotic expression vector was constructed with both the mutation of glycosylationsites N181/N197and the lack of GPI.The IRES expression of PrP proteinwas detected after the PrP vector was transfect into 293 T cell and H1-HeLacell. The results show that many forms of PrP protein are found than in eukaryotic cells. Dual luciferase expression vector with suspected PrP IRES sequence were constructed to determine possible PrP mRNA IRES areas. In the mean time, a new Western blotting methods was constructed to further confirm PrP mRNA IRES. Combined with GGG complementary sequence of 18S ribosome helix 26 region, we found PrP mRNA IRES regions were located respectively on among 197-324bp,400-459bp,496-612 bp,613-636bp and 637-693 bp. ER stress activator, Tunicamycin not Thapsigargin and Brefeldin A, induced PrP mRNA IRES protein expression. Both IRE I and PERK not ATF6 modulated PrP mRNA IRES by inhibiting three important ER stress signal passways. The results will play some roles in the mechanism of both Prion protein expression and prion disease.Fever is one of clinical manifestations of virus disease.Specific IgM detection in serum is an important means of laboratory diagnosis of viral diseases. In order to explore viral infection in febrile patients in Anhui fuyang and ZheJiang Ningbo,45 serum of hospitalized Hand foot and mouthdisease (HFMD) children from Fuyang, Anhui and 105 serum of fever patient from NingBo, Zhejiang in 2014-2015 were colleceted, serum IgM antibodies of 6 viruseswere detected and analyzed in this reserach.HFMDis common viral disease in children in many countries. HFMD can be caused by many entervirus, e.g. Coxsacklevirus-A(CVA), Coxsacklevirus-B (CVB),et al. Enterovirus 71(EV71) pathogen is the most common cause of HFMD. To study co-infected with other viruses, we collected forty-fiveserum samples of HFMD hospitalized cases according to the diagnostic criteria of HFMD from November 2013 to December 2013 in Fuyang, Anhui. Serum IgM of ADV, RSV, B19, HCMV, HPIVs, and HSV1 were detected with ELISA method. Results show that serumIgM positive of thirty cases (67.7%) ADV,8 cases (17.8%) RSV,8 cases (17.8%) parvovirus B19,3 cases (6.7%) HCMV,1 case (2.2%) HSV were found in these HFMD samples from November 2013 to December 2013 in Anhui. No IgM positive caseof HPIVs was detected in these samples. Two or two more viruses IgM positive in HFMD were also found in this study. We think that HFMD cases co-infected with many viruses including ADV, RSV, and B19 in FuYang in the 2013 winter.In order to understand parvovirus B19 infectionin Ningbo fever patients in 2014-2015, 105 serum samples were collected. Serum IgM of B19 virus was detected by ELISA. parvovirus B19 was typed through specific PCR detection. IgM antibodies of AD, HSV1, Cox, HCMV,and RSV viruses also was detected in B19 positive serum samples by ELISAKits. The results showed32 positive cases were found in Ningbo, Zhejiang in June,2014-June,2015. Positive cases gradully increased from June,2014 and peaked to 21 cases between July and August in 2014, then decreased in 2015. The results suggest that parvovirus B19 outbroke in the 2014.The type III of B19 virus was identified through PCR. At the same time, the IgM positive cases of ADV, RSV, HCMV were also foundinthese B19 IgM positive serum. |
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