Large DNA viruses normally have complex structures with many of protein components derived from both viral and host origins. The development in proteomics, especially mass spectrometry identification techniques provide powerful tools for analyzing large viruses. In this review, we have summarized the recent achievements on proteomic studies of large DNA viruses, such as herpesvirus, poxvirus, nimavirus and baculoviruse. The proteomics of baculovirus occlusion-derived virions (ODV) were emphasized. Different mass spectrometry techniques used on ,carious baculoviruses were introduced, and the identified structurally associated proteins of baculoviruses are summarized.

Objective To discuss the clinical application of simplified neonatal critical illness score (sNCTS)in comparison with original neonatal critical illness score(oNCIS)published in 2001.Method A total of 705 neonates referred to neonate ICU(NICU)from 1 st January 2007 to 31th December 2009 were prospectively studied with control.The patients were scored by oNCIS on admission day,3rd,7th days after admission and on the day of discharge or dead.At the first scoring on admission,2 items of the PaO2 and pH were excluded from oNCIS's 10 items,and the remaining 8 items were used.Three items of plasma sodium,potassium and creatinine or BUN were scored out from 8 items and the still remained 5 items were used for the subsequent 3 scorings.The remaining 8 and 5 items were used as a simplified neonatal critical illness score.The simplified NCIS was evaluated by comparing the patients'condition that was assessed by the originat NCIS.The consistency rate between oNCIS and sNCIS should be over 80%.Results There were 8 items were used to evaluate the severity of disease on admission, and the consistence rate was 86.7%with the original NCIS.The 5 items selected from the original NCIS were used on the 3rd,the 7th days and the day of discharge or death.the consistence rate with original NCIS were 86.6%to 95.7%.A close correlation existed between the original NCIS and simplified NCIS(P<0.01).There were significant differences in mortality rates of the non-serious,serious and extremely serious groups between original NCIS and simplifled NCIS used for the first and the last seorings(P<0.01).Within a cohort of neonates with the same general condition,the assessments of mortality rates were not significantly different between these two methods (P>0.05).Conclusions Compared with the original NCIS.the simplified NCIS is consistent to a large extent in disease assessment,which is a concise way to evaluate the critical ill neonates objectively and can be easily applied to clinical practice.

The etiological agent of severe acute respiratory syndrome (SARS) was identified as a new coronavirus, termed SARS-CoV. Establishment of an efficient and sensitive diagnostic system of SARS-CoV genetic materials is crucial for SARS control. In this study, we quantified SARS-CoV mRNAs in both infected cell culture lysate and in supernatant by using Real-time quantitative revere transcription-PCR based on EvaGreen鈩?dye and Taqman-MGB probes. For extensive evaluation of sensitivities and specificities, 13 pairs of primers and 4 probes were designed based on different genes of SARS-CoV. Glyceraldehydes-3-phosphate dehydrogenase (GAPDH) was selected as the internal control gene. Results showed that S-gene-specific PCR was the most sensitive for detection, but because of its sequence variability in the different viral strains, primers and a probe based on the N gene were suitable substitutions. Meanwhile, we found the mRNA concentrations in cell culture lysates were much higher than in cell supernatant and facilited more sensitive detection of the SARS-CoV.

Objective To investigate the inhibitory effect of bone marrow mesenchymal stem cells ( BMMSC) on the inflammatory response of microglial cells .Methods The samples were divided into four groups .Group I:microglial (BV-2) cells were cultured in DMEM (high glucose).Group II: BV-2 cells were cultured in DMEM containing lipopo-lysaccharide (LPS).Group III:BV-2cells and BMMSCs were co-cultured in DMEM.Group IV: BMMSCs were cultured in DMEM containing LPS .The growth state and ultrastructure of BV-2 cells were observed and the changes of TNF-αex-pression were detected .Results Different cell densities of BV-2 cells were observed under the optical microscope in an or-der from high level to low level:group I >group III >group II.The expressions of TNF-αwere:group Ⅱ >groupⅢ>group Ⅰ.Ultrastructural observation of BV-2 cells showed that there were a large number of swollen mitochondria and endoplasmic reticulum , some of them showed vacuolization .No BV-2 cells with multiple hucleoli were found in the group II indicating the absence of active cell growth .At the same time, cytolysis was observed only in the group II .The growth of BV-2 cells in the group III was better than that in the group II .Conclusions BMMSCs can inhibit inflammatory response of microglial cells, therefore, play a neuroprotective role.

Objective To investigate the correlation between red cell distribution width RDW) and prognosis in patients with severe traumatic brain injury.Methods A total of 264 consecutive patients with severe traumatic brain injury admitted from May 2012 to November 2014 were enrolled.The patients were divided into low-RDW group (RDW ＜ 15％,n =198) and high-RDW group (RDW ≥ 15％,n =66) based on their RDW levels.Between-group differences were evaluated on general conditions,acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ),inflammatory markers [high sensitivity C-reactive proteins(hs-CRP) and arterial lactate],liver/renal dysfunction,and 28-day mortality.Spearman correlation analysis of RDW with mortality was conducted.Independent factors of 28-day mortality were identified using multivariate Logistic regression.Kaplan-Meier 28-day survival curve was analyzed and survival probability of the two groups was compared using Log-Rank test.Results The 28-day mortality was significantly enhanced in high-RDW group compared to that in lowRDW group (43.9％ vs 26.8％,P ＜ 0.01).RDW ≥ 15％ related positively to APACHE Ⅱ and mortality(r =0.172 and 0.253 respectively,P ＜ 0.01),but negatively to Glagow coma score (GCS) (r =-0.169,P ＜0.01).RDW≥ 15％ was the independent risk factor for predicting the 28-day mortality (OR =2.144,95％ CI 1.202-3.826,P ＜0.01).After adjusted gender,age,and other relative factors,RDW≥15％ was still strongly correlated with the 28-day mortality(OR =2.244,95％ CI 1.076-4.678,P ＜ 0.05).Significantly lower 28-day survival rate was found in high-RDW group than in low-RDW group (P ＜ 0.01).Conclusions RDW level rises beyond the normal range on admission in patients with severe traumatic brain injury,which is closely correlated with the 28-day poor outcome.RDW≥ 15％ has significant predictive value in the prognosis.

BACKGROUND:A large number of experiments have shown that bone marrow mesenchymal stem cels (BMSCs) have good effects on the treatment of lung disease or improvement of lung injury, and its therapeutic effect is mainly oriented to reduce the inflammatory response. OBJECTIVE: To study the effects of BMSCs combined with reduced glutathione in murine models of bleomycin-induced lung injury. METHODS:BMSCs were isolated from a male NOD/SCID mouse and cel morphology and phenotype were observed. Sixty-four female NOD/SCID mice were randomly divided into a control group, a model group, a BMSCs group and a BMSCs+reduced glutathione group (combined group). The control group received intratracheal injection of normal saline, the model group received intratracheal injection of bleomycin, and the BMSC group received BMSCs injectionvia the tail vein at 2 hours after intratracheal injection of bleomycin, and combined group received BMSCs+reduced glutathione injection via the tail vein at 2 hours after intratracheal injection of bleomycin. Al mice were kiled after 7 days, and levels of tumor necrosis factor-α, interleukin-β, malondialdehyde in lung tissue were detected and lung tissue specimens were obtained for pathological examination. RESULTS AND CONCLUSION:BMSCs were fibroblast-like cels, Positive for CD10, CD13 and CD44, but negative for CD34 and CD45. Compared with the control group, the levels of tumor necrosis factor-α, interleukin-β and malondialdehyde were increased in the model group, while decreased significantly in the BMSCs group and combined group (P < 0.05), especialy in the combined group (P < 0.05). Pathological examination showed that more serious lung injury was observed in the model group and BMSCs group than the combined group. These findings indicate that BMSCs combined with reduced glutathione can more effectively protect against bleomycin-induced lung injury.

Purpose To assess the effect of pelvicfloor electrical stimulation treatment by transperineal sonography, so as to provide guidance for postpartum pelvic floor rehabilitation.Materials and Methods Thirty-one delivery women who received postpartum pelvicfloor rehabilitation were selected as research group and 30 non-pregnant young women as control group in the study. Pelvicfloor electromyography treatment was taken in research group, and transperineal sonography was undergone in quiet state and pressure state (Valsalva operation) before treatment, after the fifth treatment and after the tenth treatment respectively to measure the bladder neck to the public symphysis edge distance (Dr, Ds), bladder neck angle (θr,θs) and bladder rotation angle (θ). A comparison was made between the research group and control group.Results①Dr and Ds in research group were both lower than those in control group (P<0.05), butθr,θs andθ were all higher than those in control group (P<0.05).②Dr and Ds showed a gradual increase (P<0.05) whilstθr presented a gradual decrease before the treatment, after the fifth treatment and after the tenth treatment (P<0.05).Conclusion Natural childbirth may damage the function of pelvicfloor; electrical stimulation of pelvic therapy is effective on postpartum pelvicfloor rehabilitation; and transperineal sonography can objectively assess the effect of this therapy.

As a protein expression vector,the baculovirus demonstrates many advantages over other vectors.With the development of biotechnology,baculoviral vectors have been genetically modified to facilitate high level expression of heterologous proteins in both insect and mammalian cells.These modifications include utilization of different promoters and signal peptides,deletion or replacement of viral genes for increasing protein secretion,integration of polycistronic expression cassette for producing protein complexes,and baculovirus pseudotyping,promoter accommodation or surface display for enhancing mammalian cell targeting gene delivery.This review summarizes the development and the current state of art of the baculovirus expression system.Further development of baculovirus expression systems will make them even more feasible and accessible for advanced applications.

Influenza virus is a continuous and severe global threat to mankind.The continuously re-emerging disease gives rise to thousands of deaths and enormous economic losses each year,which emphasizes the urgency and necessity to develop high-quality influenza vaccines in a safer,more efficient and economic way.The influenza subunit and VLP vaccines,taking the advantage of recombinant DNA technologies and expression system platforms,can be produced in such an ideal way.This review summarized the recent advancements in the research and development of influenza subunit and VLP vaccines based on the recombinant expression of hemagglutinin antigen (HA),neuraminidase antigen (NA),Matrix 2 protein (M2) and nucleocapsid protein (NP).It would help to get insight into the current stage of influenza vaccines,and suggest the future design and development of novel influenza vaccines.

Objective:To determine the inflammatory factor levels and inflammatory cell percentages in the patients with cough variant asthma (CVA ), and to clarify their potential role in the pathogenesis of CVA unresponsive to bronchodilator treatment.Methods:60 patients with CVA were randomly selected and divided into CVA unresponsive to bronchodilator treatment group (n=30)and CVA responsive to bronchodilator treatment group (n=30).As the same time 30 cases of healthy persons were used as normal control group.The levels of interluekin-8 (IL-8)and esoinophil cationic protein (ECP)in their induced sputum were detected,the classification of inflammatory cells in their induced sputum were observed, and their scores of cough symptom were recorded. Results:The IL-8 level in the induced sputum of the patients in CVA unresponsive to bronchodilator treatment group was higher than that in CVA responsive to bronchodilator treatment group and normal control group (P<0.05).The ECP level in the induced sputum of the patients in CVA unresponsive to bronchodilator treatment group was lower than that in CVA responsive to bronchodilator treatment group (P<0.05),but it was similar to the level in normal control group (P>0.05).The neutrophil percentages in the induced sputum of the patients in CVA unresponsive to bronchodilator treatment group were higher than those in CVA responsive to bronchodilator treatment group and normal control group (P<0.05).The scores of cough symptom of the patients in CVA unresponsive to bronchodilator treatment group was positively correlated with IL-8 level (r=0.764,P<0.01), and the scores of cough symptom of the patients in CVA unresponsive to bronchodilator treatment group was positively correlated with the neutrophil percentage in induced sputum (r=0.889,P<0.01).Conclusion:IL-8 and neutrophil may be associated with the incidence of CVA unresponsive to bronchodilator treatment. They can aggravate the inflammation and hypersensitivity of airway and cough symptom. The determination of IL-8 and neutrophil can be used as an accessory method in the diagnosis and j udgement of severity degree and curative effect of CVA in clinic.