Animals ; Apoptosis ; Cell Cycle ; Cell Movement ; Cell Proliferation ; Connexin 43 ; genetics ; metabolism ; Down-Regulation ; Gap Junctions ; metabolism ; physiology ; Humans ; Neoplasms ; metabolism ; pathology ; Tumor Suppressor Proteins ; genetics ; metabolism

Objective To investigate the difference in ocular surface microbiota between dry eye patients and healthy subjects,and discuss the role of microbiota in dry eye.Methods Twenty cases of dry eye patients and 90 cases of healthy subjects were collected in the PLA General Hospital and Zhongshan Ophthalmic Center.The samples of conjunctiva impression cytology were collected from all subjects,and then metagenomic shotgun sequencing was performed following the DNA extraction.The differences in alpha diversity and metabolic pathways of the ocular surface microbiota between dry eye patients and healthy subjects were evaluated.Results There was no significant difference in alpha diversity of the microbial community between dry eye patients and healthy subjects (P =0.13).However,an increase of 15 species and a decrease of 10 species were detected on the ocular surface of dry eye patients.The enriched antibiotic resistance genes in dry eye patients were more than healthy subjects.Conclusion Although the alpha diversity of the microbial community on ocular surface between dry eye patients and healthy subjects are not distinguishable,a significant difference could be found in relative abundance and metabolic pathways,suggest that these specific microbiome may be related to the pathogenesis and disease progression of dry eye.

Female ; Humans ; Infant ; Lung ; physiopathology ; Male ; Pneumonia, Viral ; physiopathology ; Respiratory Function Tests ; Respiratory Syncytial Virus Infections ; physiopathology ; Respiratory Syncytial Viruses

ObjectiveTo observe the therapeutic effect and prognosis of nimodipine on severe traumatic brain injury. Methods64 patients with severe traumatic brain injury were divided into the nimodipine group(32 cases) and the routine treatment group(32 cases). The Glasgow Coma Scale(GCS) was assessed before and after the treatment. The Activities of Daily Living(ADL) and cognitive ability were evaluated in clear-headed patients. After 6 months follow-up, the Glasgow Outcome Scale (GOS), Bathel index and Mini Mental State Examination (MMSE) score were carried on.ResultsIn both of the two groups, GCS score were increased distinctively after treatment. The MMSE score in nimodipine group was higher than that of routine treatment group. There was no statistic difference in GOS and ADL between two groups after 6 months, but MMSE in nimodipine group was higher than that of routine treatment group. Conclusions Nimodipine could be helpful in cognitive function. The prognosis of severe traumatic brain injury lied on the degree of cerebral damage.

Middle East respiratory syndrome coronavirus (MERS-CoV) was identified as a novel human coronavirus and posed great threat to public health world wide,which calls for the development of effective and safe vaccine urgently. In the study, peptide epitopes tagrgeting spike antigen were predicted based on bioinformatics methods. Nine polypeptides with high scores were synthesized and linked to keyhole limpet hemocyanin (KLH). Female BALB/C mice were immunized with individual polypeptide-KLH, and the total IgG was detected by ELISA as well as the cellular mediated immunity (CMI) was analyzed using ELIs-pot assay. The results showed that an individual peptide of YVDVGPDSVKSACIEVDIQQTFFDKTWPRPIDVSKADGI could induce the highest level of total IgG as well as CMI (high frequency of IFN-γ secretion) against MERS-CoV antigen in mice. Our study identified a promising peptide vaccine candidate against MERS-CoV and provided an experimental support for bioinformatics-based design of peptide vaccine.
Animals ; Antibodies, Viral ; immunology ; Computational Biology ; Coronavirus Infections ; immunology ; prevention & control ; virology ; Female ; Humans ; Immunization ; Mice ; Mice, Inbred BALB C ; Middle East Respiratory Syndrome Coronavirus ; genetics ; immunology ; Peptides ; administration & dosage ; genetics ; immunology ; Spike Glycoprotein, Coronavirus ; administration & dosage ; genetics ; immunology ; Viral Vaccines ; administration & dosage ; genetics ; immunology

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.

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 effect of smoking and smoking cessation on the phosphorylation of IKK-β in type 2 diabetic rats. Methods Forty-two six-week-old Wistar rats were randomly divided into 4 groups: normal control(NC, n =7), diabetes control (DC, n =7), diabetes with smoking (DS, n = 14) and diabetes with smoking cessation(SC, n = 14). Rats in DS and SC groups were further assigned randomly into 8w and 12w subgroups. DS group was given passive smoking twice a day for 8 or 12 weeks, while SC group ceased passive smoking for 4 weeks after 8 or 12 weeks of smoking . Western blot method was used to detect the level of IKK-13 phosphorylation in skeletal muscle. Results Compared with the NC group,the phosphorylation of IKK-β protein in DC group was increased (0. 16±0. 05 vs 0. 30±0. 08, P < 0. 01). There was an increasing trend with the phosphorylation level of IKK-β in the DS (8w) subgroup, but there was no statistical difference between the DC group and SC(8w) subgroup (0. 40±0. 09 vs 0. 30±0. 08,0. 36±0. 10, P >0. 05). The phosphorylation level of IKK-β in DS(12w) group increased obviously, being significantly higher than that in the DC group and SC (12w) subgroup(0. 74 ± 0. 11 vs 0.30±0.08,0.35±0.07,P < 0.01). Conclusion With the prolongation of smoking duration, the phosphorylation of IKK-β in type 2 diabetic rats increased. After smoking cessation, the phosphorylation of IKK-β decreased. The phosphorylation of IKK-β may be involved in the mechanism by which smoking causes type 2 diabetes.

Objective To investigate the effect of human cytomegalovirus (HCMV) on proliferation of colony forming unit T-lymphocyte (CFU -TL)and its interference methods. Methods Normal CFU - TL culture was used as blank control. Normal CFU- TL culture system plus inactivated HCMV fluid as inactivated HCMV control. The dilution of 1:10,1:100,1:1000 were added into CFU -TL colonies culture system directly as infected group. Astragalus (AMI) and ganciclovir(GCV) were added into culture system with HCMV dilution of 1:10 as experimental group. By methylcellulose semi-solid culture, different concentrations of HCMV - AD1699 affect CFU-TL and interfered by astragalus AMI, GCV. CFU - TL were surveyed. The effect of HCMV on CFU-TL proliferation was measured by MTT; HCMV-AD169 DNA in CFU-TL was found by PCR. Results 1. Compared with control group, the numbers of CFU -TL in the HCMV infection groups decreased significantly(P

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.