1.Virion Proteomics of Large DNA Viruses
Ranran WANG ; Zhihong HU ; Hualin WANG ; Fei DENG
Virologica Sinica 2009;24(4):268-284
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.
2.Evaluation of the clinical application of simplified neonatal critical illness score
Fei WANG ; Fenghua HU ; Guowei SONG ; Xiaoxu REN
Chinese Journal of Emergency Medicine 2011;20(5):469-472
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.
3.Evaluation of Sensitivities and Specificities of SARS-CoV Detection by Real-time Quantitative Reverse Transcription-PCR Assays
Lili XU ; Zhihong HU ; Hualin WANG ; Xiao HAN ; Fei DENG
Virologica Sinica 2009;24(3):187-193
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.
4.Therapeutic Effect of Pelvic Floor Rehabilitation on Electrical Stimulation Assessed by Transperineal Sonography
Ting HU ; Fei XIA ; Hong XU ; Juan WANG ; Xinxian GU
Chinese Journal of Medical Imaging 2014;(11):849-852
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.
5.Genetic Modification of Baculovirus Expression Vectors
Shufen LI ; Hualin WANG ; Zhihong HU ; Fei DENG
Virologica Sinica 2012;27(2):71-82
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.
6.Developments of Subunit and VLP Vaccines Against Influenza A Virus
Maping DENG ; Zhihong HU ; Hualin WANG ; Fei DENG
Virologica Sinica 2012;27(3):145-153
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.
8.Diagnostic Value of 64-MSCT and 1.5T MRI in Nurseling With Congenital Vascular Rings
Gengwu LI ; Chang WANG ; Jun HU ; Xu LI ; Weimin FEI
Chinese Circulation Journal 2016;31(2):156-160
Objective: To investigate the diagnostic value of 64 multislice spiral CT (64-MSCT) and 1.5T MRI in nurseling with congenital vascular rings.
Methods:We retrospectively studied the diagnostic records of 64-MSCT and 1.5T MRI in 48 nurseling with congenital vascular rings. There were 42 cases received CTA, 12 received MRA and 6 received both CTA and MRA. We classiifed the types of vascular rings by Edwards sketch map of aortic arch embryonic development and trachea wrapping conditions. The diagnostic values of 64-MSCT and 1.5T MRI on vascular rings were compared.
Results: There were 25 patients with complete vascular rings including 3 of double aortic arch, 12 right aortic arch with left subclavian artery and patent ductus arteriosus (PDA), 9 combining with arterial ligament, 1 neck right aortic arch with left subclavian artery combining left PDA;and 23 patients with in-complete vascular rings including 2 pulmonary artery sling, 9 left aortic arch with right subclavian artery combining left PDA, 10 combining with arterial ligament, 1 right aortic arch with left subclavian artery combining right PDA, 1 with innominate artery compressed trachea. There were 32/48 patients combined with cardiac malformations, both 64-MSCT and 1.5T MRI may well reveal the composition and special relationship of vessel rings. The minimum intensity projection (MinIP) and air volume rendering (VR) of 64-MSCT may clearly display the morphology and development of trachea, image fusion technique could directly show the special relationships among vascular rings, trachea and esophagus. While 1.5T MRI was relatively poor to present tracheal image.
Conclusion: 64-MSCT could accurately and comprehensively display the special relationships among the vessels rings, trachea and esophagus;which was the best method for diagnosing the nurseling with vessel rings in clinical practice.
9.Clinical effects and safety of catheter-directed thrombolysis for acute lower extremity deep vein thrombosis in patients aged 70 years and over
Qianyi CHEN ; Hua WANG ; Xiaoman HU ; Zhewei FEI ; Jingmin OU
Chinese Journal of Geriatrics 2016;35(2):133-136
Objective To analyze the efficacy and safety of catheter-directed thrombolysis (CDT) for acute lower extremity deep venous thrombosis (DVT) in patients aged 70 years and over.Methods Clinical data of 109 cases who had acute lower extremity DVT and had been treated with CDT from March 2011 to September 2014 were retrospectively analyzed.Results Inferior vena cava filters (IVCF) were implanted in 109 patients.A thrombolytic catheter was inserted from the contralateral femoral vein (21 cases),ipsilateral popliteal vein (44 cases),posterior tibial vein (28 cases) or small saphenous vein (16 cases).The duration of thrombolysis was (5.47±2.13) d.The dosage of urokinase was (3.80 ± 0.56) million units.Forty-two cases underwent balloon dilatation alone,and 67 cases combined balloon dilatation with stent implantation.Gingival bleeding occurred in 7 patients and gross hematuria occurred in 4 patients during thrombolysis,which disappeared after medication adjustment.No symptomatic pulmonary embolism (PE) or other serious complications were found in any patients.There were significant differences in the venous patency score and lower limb circumferences at 10 cm above and below the knee one week after treatment (t=3.874,P=0.031).Ninety-seven cases were followed up for a mean period of (20.76.5) months,and the vein patencyrate was (77.9± 10.5) % and (73.1±9.4) % at 6 and 18 months after treatment,respectively.Recurrence of deep vein thrombosis was found in 6 cases,of which 2 cases had recurrent stent thrombosis,and treatment with CDT again was successful.No severe deep venous thrombosis syndrome (PTS) was found during the follow-up.Conclusions CDT is a minimally invasive procedure and can rapidly resolve limb venous drainage disorders.CDT is safe,has few complications and usually generates satisfactory outcomes.For very-old elderly patients who have no anticoagulation and thrombolytic contraindications,CDT is a useful option for the treatment of acute lower limb DVT.
10.Bone marrow mesenchymal stem cells inhibit inflammatory response of microglial cells
Kaihua ZHANG ; Fei WANG ; Huimin HU ; Hua HUANG ; Yihua AN
Acta Laboratorium Animalis Scientia Sinica 2014;(2):1-5
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.