Base Sequence ; Hepacivirus ; drug effects ; genetics ; Hepatitis C ; drug therapy ; virology ; Humans ; Molecular Sequence Data ; Oligonucleotides ; therapeutic use ; Phosphorothioate Oligonucleotides ; therapeutic use

In addition to the structural genes of the coronavirus genome, S, E, M, and N, there are several additional genes called "group-specific or accessory genes". Their gene products are designated as "accessory proteins", as reports to date make it clear that these proteins are not essential for virus replication in vitro. Nevertheless, many of these genes are still maintained in the virus genome under selective pressure, suggesting that they might play a very important role in the survival of the virus in the natural environment of the infected host. This review will summarize the research progress in the functions of coronavirus accessory genes.
Animals ; Coronavirus ; genetics ; metabolism ; Coronavirus Infections ; virology ; Humans ; Open Reading Frames ; Viral Proteins ; metabolism

Animals ; Autophagy ; Coronaviridae ; genetics ; pathogenicity ; physiology ; Coronaviridae Infections ; physiopathology ; virology ; Humans ; Virus Replication

Hepatitis C virus (HCV) accounts for the majority of cases of transfusion acquired hepatitis and may cause chronic hepatitis, cirrhosis and hepatocellular carcinoma. Currently, there is no vaccine against HCV and treatment is expensive and not always effective.The adaptive host immune response in viral clearance of HCV infection was described and the recent progress in vaccine development of HCV, focusing on the fields of DNA vaccine candidates, recombinant viral vectored vaccine candidates and combined (prime-boost) vaccine candidates were summarized. Some challenges and limitations of developing a HCV vaccine were also analysed. In summary, a promising approach of developing an experimental HCV vaccine to induce extremely potent and broad T-cell responses based on prime-boost strategy was presented.

Objective To determine the clinical value of EUS in combination with detection of CA19-9 and CA242 for early diagnosis of pancreatic cancer.Methods General features of high-risk group were determed by EUS and the plasma levels of CA19-9 and CA242 measured by ELISA.Results The serum level of CA19-9 was abnormal in 9.80% (10/102) and that of CA242 in 12.75% (13/102) of the patients.They were both abnormal in 7 cases.Of the 7 cases, 3 were found to suffer from pancreatic cancer by EUS, 1 had the tumor that had been clinically confirmed, 1 had posterior peritoneal tumor and 2 were normal.Among 102 cases detected by EUS, 2 showed low echo in pan-creatic head, 2 semi-cyst, 3 pancreatic cancer and proved by pathology.Accurate diagnotic rate, sensi-tivity and specificity were 90.20% (92/102), 50% (4/8) and 93.62% (88/94), respectively, for serum CA19-9.For the serum CA242, the 3 parameters were 87.25% (89/102), 50% (4/8) and 90.43% (85/94), respectively.For combination of both methods, they were 98.04% (100/102), 100%(8/8) and 97.87%(92/94), respectively.Accurate diagnostic rate of the latter was significantly higher than that of the former 2(P＜0.05).Concision Combination of EUS with detection of serum CA19-9 and CA242 is of great vlaue for early screening of pancreatic cancer.

Objective To investigate the immunogenicity and cross protective effects of two novel HCV DNA vaccines in a mice model.Methods Two self-replicating alphavirus vector-based HCV DNA vaccines, pSCK CE1E2Y and pSCK H155, were constructed based on the genes encoding the structural pro-teins (Core, E1 and E2) and structural and NS3 fusion proteins (Core, E1 , E2 and NS3) of a HCV strain isolated from a Chinese patient (genotype 1b, Hebei strain), respectively.Western blot analysis was per-formed to detect the expression of fusion antigens.The BALB/c mice were intradermally immunized with the recombinant DNA vaccines by using electroporation.The immune responses induced in mice and the cross protective effects of the recombinant DNA vaccines were evaluated.Results The DNA vaccines effectively expressed the target antigens in vitro.The antigen-specific antibody responses and specific T cell immune re-sponses were induced in mice by the immunization of replicative DNA vaccines.However, no effective cross protection was provided by either of the DNA vaccines in the surrogate challenge model based on a recombi-nant heterologous HCV (JFH1, 2a) vaccinia virus strain.Conclusion Although no effective cross protec-tion was observed, both of the two replicative DNA vaccines could induce strong humoral and cellular im-mune responses against multi-target antigens of HCV strains.This study has paved the way for further inves-tigation on the development of novel HCV vaccines.

Objective To evaluate the expression of macrophage migration inhibitor factor (MiF) and cyclinD1 in pancreatic carcinoma and their relationships with clinical pathology characteristics. Methods The expression of MIF and eyclinD1 in 89 carcinoma and 5 normal pancreatic tissues was detected with immunohis-tochemistry methods, and the relationships among MIF and cyclinD1 expression and clinicopathological factors were studied. Results The overexpression of MIF and cyclinD1 was found in 88.8%, and 50. 6% of pancre-atic carcinoma tissues respectively. The overexpression of MIF had a significant correlation with Ⅰ,Ⅱ,Ⅲ,Ⅳ tumor stage (69. 2%, 94. 7%, 96. 4%, 100%, P <0.05), while the positive expression rate of cyclinD1 only had a significant correlation with tumor stages Ⅲ,Ⅳ (33. 3%, 68. 8%, P <0. 05). Both of the two proteins had a correlative tendency with pathological grade and lymph node metastasis. The different expression of MIF between pancreatic carcinoma with and without liver metastasis had no statistical significance, (100% ,85.9%, P >0. 05)while there was a statistically significant difference about cyclinD1 (66. 7% ,46. 5% ,P <0. 05). A significant positive correlation was also found between MIF and cyclinD1 (P < 0. 05). Conclusion The ex-pression of MIF and CyclinD1 was higher in pancreatic cancer tissues than in normal tissue, and they may be associated with the malignant stage, tumor differentiation, local lymph node and liver metastasis of this tumor.

Objective To assess the left ventricular systolic asynchronicity in chronic ischemic model with real-time three-dimensional echocardiography (RT-3DE), and to explore the affection of low-dose dobutamine to it. Methods A chronic ischemic model was induced by placing an Ameroid constrictor in the left circumflex(LCX) in swines,then full volume RT-3DE was performed by Philips iE33 with X3-1 probe combining rest and stress(dobutamine stress echocardiography, DSE) every week after LCX constriction.Ten normal pigs before operation served as controls (group A). Examination of all the models post operation were grouped into group B (mild stenosis, LCX stenosis＜50% ), group C (moderate stenosis, LCX stenosis 50%～75%) and group D (severe stenosis, LCX stenosis≥75%) according to the results of coronary angiography. Images were copied to QLAB 5.2 postprocess workstation,and 3DQA software was used to analyze the full volume data sets. The time to the point with minimal systolic volume (Tmsv) in each segment was taken to derive the following indexes of systolic synchrony: the maximum difference of Tmsv (Tmsv-dif) and standard deviation(Tmsv-SD) among various segments and standard index (Tmsv-dif% and Tmsv-SD%), to evaluate left ventricular dyssynchrony. Tmsv3-6 represented the maximum difference of Tmsv between lateral segment and posterior septum (Tmsv3-5: between lateral segment and inferior) in basal level. Results Tmsvl2-Dif%, Tmsv6-Dif%, Tmsv3-6% and Tmsv3-5% under stress condition in group C and D were significantly higher than those at rest;all the data in group D were significantly higher than in group A and B, and in group C higher than group A ( P ＜0.05,0.01 ). Compared with group A,Tmsv6-Dif,Tmsv3-6 and Tmsv3-5 in group B were significantly increased under stress condition,and so did their standardize data under both rest and stress conditions ( P ＜ 0.05, 0. 01 ). Conclusions RT-3DEcombined with DSE could display sensitively the left ventricular asynchrony caused by chronic ischemia,and that will be more significant in lateral wall in LCX stenosis than in normal segments.

To develop a safe and broad-spectrum effective hepatitis C virus (HCV) T cell vaccine,we constructed the recombinant adenovirus-based vaccine that carried the hepatitis C virus truncated NS3 and core fusion proteins. The expression of the fusion antigen was confirmed by in vitro immunofluorescence and western blotting assays. Our results indicated that this vaccine not only stimulated antigen-specific antibody responses,but also activated strong NS3-specific T cell immune responses. NS3-specific IFN-γ+ and TNF-α+ CD4+ T cell subsets were also detected by a intracellular cytokine secretion assay. In a surrogate challenge assay based on a recombinant heterologous HCV (JFH1,2a) vaccinia virus,the recombinant adenovirus-based vaccine was capable of eliciting effective levels of cross-protection. These findings have im- portant implications for the study of HCV immune protection and the future development of a novel vaccine.
Adenoviridae ; genetics ; metabolism ; Animals ; CD4-Positive T-Lymphocytes ; immunology ; Cross Protection ; Female ; Genetic Vectors ; biosynthesis ; genetics ; Hepacivirus ; genetics ; immunology ; Hepatitis C ; immunology ; prevention & control ; virology ; Humans ; Interferon-gamma ; immunology ; Mice ; Mice, Inbred BALB C ; Recombinant Proteins ; administration & dosage ; genetics ; immunology ; Viral Core Proteins ; administration & dosage ; genetics ; immunology ; Viral Hepatitis Vaccines ; administration & dosage ; genetics ; immunology ; Viral Nonstructural Proteins ; administration & dosage ; genetics ; immunology

Objective To investigate the development strategy of novel T cell based vaccine against HCV infection.Methods BALB/c mice were primed with pSCK-based DNA vaccine and boosted with type 5 adenoviral vector-based vaccine, which expressed the structural proteins ( Core, E1 and E2) de-rived from a Chinese HCV patient (genotype 1b, Hebei strain).Enzyme linked immunospot assay (ELIS-POT) and intracellular cytokine staining ( ICS) were used to analyze the elicited antigen-specific immune re-sponses and the efficacy of cross-protection.Results Immunization of mice with the prime-boost vaccination strategy elicited stronger T cell immune responses against multiple HCV antigens than using the DNA vac-cines alone, especially the IFN-γ-secreting T cell responses against E1 protein as indicated by ELISPOT as-say.ICS data indicated that the prime-boost regimen elicited more TNF-α-producing CD4+and IFN-γ-produ-cing CD8+T cells against E1 protein and high levels of IFN-γ-producing CD4+and CD8+T cells against E2 protein in comparison with immunization with DNA vaccines.Moreover, the prime-boost vaccination was ca-pable of eliciting effective cross-protection in a surrogate challenge model based on a recombinant heterolo-gous HCV (JFH1, 2a) vaccinia virus.Conclusion The prime-boost vaccination using DNA and rAd5-based vaccine expressing HCV structural antigens induced significant cellular immune response and cross-protection in mice, suggesting the possibility of using it as a promising T cell based vaccine against HCV in-fection.