1.The relationship between oxidative injury induced by low glucose and mitochondrial membrane potential in HUVEC-12 cells
Wen LU ; Yaoming XUE ; Bo ZHU ; Xin LIAN ; Ning LIU
Chinese Journal of Internal Medicine 2011;50(10):873-876
ObjectiveTo investigate the relationship between the oxidative injury induced by low glucose and mitochondrial membrane potential in HUVEC-12 cells. Methods Human umbilicalvein endothelial cells HUVEC-12 were cultured in low concentration glucose for 4 h.Cell viability of HUVEC-12 cell was assessed with MTT assay.Dihydroethidium (DHE) was used as a reactive oxygen species (ROS)capture, which was detected the mean fluorescence intensity of samples and Rhodamine 123 as a fluorescence detector was to measure the level of mitochondrial membrane potential (MMP) in cells.Results Comparing to HUVEC-12 cells viability in 5.5 mmol/L glucose group (96.80 ±3.20)%, cells exposed to 2.8 mmol/L glucose group (66.40 ± 1.60) % and 0 mmol/L glucose group (58.93 ± 1.67) % were decreased by 32% and 40% respectively (P < 0.01).ROS level of 5.5 mmoL/L glucose group, 2.8 mmol/L glucose group and 0 mmol/L glucose group were 0.59 ± 0.02, 0.74 ± 0.04 and 0.88 ± 0.05,respectivdy, increased by 25% in cells exposed to 2.8 mmol/L glucose and by 48% in cells without glucose exposure comparing to 5.5 mmol/L glucose group (P <0.01) ; MMP levels of 5.5 mmol/L glucose group,2.8 mmoL/L glucose group and 0 mmoL/L glucose group were 148.83 ± 3.51, 271.07 ± 19.54 and357.74 ±51.32 respectively, increased to 1.8 times in cells exposed to 2.8 mmol/L glucose and to 2.4times in cells without glucose exposure comparing to 5.5 mmoL/L glucose group (P < 0.01).Conclusion Low glucose leads to injury in HUVEC-12 cells, which is probably induced by the oxidative stress via the increasing MMP.
2.Effects of ryanodine receptor channels on the spontaneous contractions of detrusor in rats of detrusor instability
Haihong JIANG ; Gensheng LU ; Qianjun WEN ; Bo SONG
Journal of Third Military Medical University 2003;0(08):-
Objective To explore the myogenic basis of the increased excitability and contractile activity in detrusor instability (DI) and investigate the differences of spontaneous contractions by ryanodine receptor (RyR) channels regulation in sarcoplasmic reticulum (SR) between DI and normal bladder muscle and of the RyR channels protein expression. Methods DI model was confirmed by filling cystometry from rats that underwent partial bladder outlet obstruction (BOO) about 8 weeks ago. Muscle strips were dissected from fresh bladder under microscope and the isometric tension in DI and normal strips were detected. The contractions were recorded in these strips exposed to some agents. SR microsome protein was obtained from DI and normal bladder muscle preparations and was used for Western blot analysis to determinate RyR channels expression. Results Treated with RyR channels blocker ryanodine , the contractile frequence significantly increased in normal strips, but not in DI muscle. Western blot analysis showed that RyR channels expression in DI muscle was significantly less than that in normal preparations. Conclusion RyR channels act a negative role in spontaneous contractile activity and the presumed mechanism may involve in Ca 2+ release of RyR channels which causes activation of Ca 2+ -dependent K + channels to decrease contractility. But this crosstalk mechanism is weaken in DI muscle, which provides a chance for spontaneous contractile overactivity.
3.Bioinformatics-based Design of Peptide Vaccine Candidates Targeting Spike Protein of MERS-CoV and Immunity analysis in Mice.
Jiaming LAN ; Shuai LU ; Yao DENG ; Bo WEN ; Hong CHEN ; Wen WANG ; Wenjie TAN
Chinese Journal of Virology 2016;32(1):77-81
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
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Antibodies, Viral
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immunology
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Computational Biology
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Coronavirus Infections
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immunology
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prevention & control
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virology
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Female
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Humans
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Immunization
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Mice
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Mice, Inbred BALB C
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Middle East Respiratory Syndrome Coronavirus
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genetics
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immunology
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Peptides
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administration & dosage
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genetics
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immunology
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Spike Glycoprotein, Coronavirus
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administration & dosage
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genetics
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immunology
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Viral Vaccines
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administration & dosage
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genetics
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immunology
4.Screening of High Daptomycin-producing Strain by He-Ne Laser Irradiation and Streptomycin Resistance Screening Method
Wen-Yu LU ; Jian-Ping WEN ; Jing-Hua FAN ; Bo-Xiang CAO ; Bing SUN ;
Microbiology 1992;0(03):-
The spores suspension of Streptomyces roseosporus D-38 irritated with 20mW He-Ne laser for 20 min were incubated on G1 medium plates containing 1. 9?g/mL of streptomycin. Ten percent of mutants increased the potency of daptomycin by streptomycin-resistance method, including the mutant LC-54, which could produce daptomycin 81. 2 mg/L, which was 39% higher than that of the beginning strain by flask fermentation.
5.Small molecular agents against MERS-CoV infection.
Xiao-yun ZENG ; Lu LU ; Shi-bo JIANG ; Shu-wen LIU
Acta Pharmaceutica Sinica 2015;50(12):1520-1526
Middle East respiratory syndrome coronavirus (MERS-CoV) has caused outbreaks of SARS-like disease with 35% case-fatality rate, mainly in the Middle East. A more severe outbreak of MERS occurred recently in the Republic of Korea, where 186 people contracted the infections, causing great concern worldwide. So far, there has been no clinically available drug for the treatment of MERS-CoV infection. The potential drugs against MERS-CoV mainly consist of monoclonal antibodies, peptides and small molecular agents. Small molecular agents have an advantage of easier synthesis, lower cost in production and relatively higher stability. There is better chance for those candidates to gain a quick development. This article reviews the progress of developing small molecular MERS-CoV agents.
Antibodies, Monoclonal
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pharmacology
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Antiviral Agents
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pharmacology
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Coronavirus Infections
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drug therapy
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Drug Design
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Humans
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Middle East Respiratory Syndrome Coronavirus
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drug effects
6.Development of peptidic MERS-CoV entry inhibitors.
Shuai XIA ; Qian WANG ; Shu-wen LIU ; Lu LU ; Shi-bo JIANG
Acta Pharmaceutica Sinica 2015;50(12):1513-1519
In 2012, a new SARS-like coronavirus emerged in the Middle East, namely the Middle East respiratory syndrome coronavirus (MERS-CoV). It has caused outbreaks with high mortality. During infection of target cell, MERS-CoV S protein S1 subunit binds to the cellular receptor (DPP4), and its S2 subunit HR1 and HR2 regions intact with each other to form a stable six-helix bundle to mediate the fusion between virus and target cell membranes. Hence, blocking the process of six-helix bundle formation can effectively inhibit MERS-CoV entry into the target cells. This review focuses on the recent advance in the development of peptidic entry inhibitors targeting the MERS-CoV S2 subunit.
Antiviral Agents
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pharmacology
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Coronavirus Infections
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drug therapy
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Dipeptidyl Peptidase 4
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metabolism
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Drug Design
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Humans
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Middle East Respiratory Syndrome Coronavirus
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drug effects
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physiology
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Peptides
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pharmacology
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Spike Glycoprotein, Coronavirus
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metabolism
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Virus Internalization
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drug effects
7.CorreIation of retinaI vein occIusion with bIood Iipids and carotid artery changes
Wen-Chao, YANG ; Fang-Fang, REN ; Xiao-Bo, LU ; Qun, FU
International Eye Science 2015;(3):489-491
· AlM: To investigate the correlation of retinal vein occlusion ( RVO ) with blood lipids and carotid artery changes.
· METHODS: Forty cases ( 40 eyes ) with RVO who presented to Eye Hospital of Xinxiang Medical University between May 2013 and April 2014 were selected as the research objects. Proceeded blood lipids and color doppler ultrasonography examination, including total cholesterol ( TC ) , triglycerides ( TG ) , high -density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol ( LDL-C ) , common carotid artery intima-media thickness, carotid plaques, internal carotid artery blood flow mechanics parameters were detected.Thirty eyes ( 30 cases ) were enrolled as control underwent above examinations.
·RESULTS:TC, TG, LDL-C of RVO group was obviously higher than those of the control group ( P<0.05), while HDL-C level was obviously lower than that of the control group ( P<0.05 ). lncidence of carotid artery plaque formation in RVO group was obviously higher than that in the control group. lntima-media thickness ( lMT ) of common carotid artery was obviously increased in RVO group (P<0.05).Both peak systolic velocity ( PSV) and end diastolic velocity ( EDV ) of internal carotid artery reduced (P<0.05), and Resistance index (Rl) increased (P<0.05).The eyes of the sick side and the contralateral carotid artery measured value had no statistical difference ( P >0.05 ) . There were also no statistical difference between ipsilateral and contralateral carotid artery measured value of control group (P>0.05).There were no differences in age, sex between RVO group and control group (P>0.05).
· CONCLUSlON: Lipid metabolism disorder, carotid artery changes is closely related to the pathogenesis of RVO.
8.Intra-operative monitoring of neuro-electrophysiology in spinal tuberculosis surgery
Yi CHEN ; Zhixiong LIN ; Wen LI ; Qi LIU ; Jingming WU ; Bo BAI ; Weijie LU
Chinese Journal of Physical Medicine and Rehabilitation 2014;36(4):287-290
Objective To investigate the efficacy of combined monitoring of motor evoked potentials with transcranial electrical stimulation (TES-MEP),somatosensory evoked potentials (SEP) and spontaneous electromyo-graphy (s-EMG) in tuberculosis surgery involving the thoracic,lumbar and sacral vertebrae.Methods Twenty-seven patients with tuberculosis of the thoracic vertebrae (T2-L2) received intra-operative SEP and TES-MEP monito-ring.Combined SEP,TES-MEP and spontaneous EMG monitoring were employed in 11 patients with tuberculosis of the lumbar or/and sacral vertebrae (L3-S1).SEP and TES-MEP were used to precisely observe the status of the sen-sory and motor pathways; s-EMG responses were used to more accurately localize nerve root irritation.ResuIts (1) SEP monitoring was successful in all of the operations.TES-MEPs were successfully monitored in 35 of them (92.1%).Combined motor and sensory monitoring was successfully achieved in 35 cases (92.1%).Abnormal SEPs were observed in 3 cases (7.9%),while abnormal MEPs were observed in 11 cases (28.9%).Abnormality in both the SEP and TES-MEP occurred in 2 cases (5.3%).There were 9 cases (23.7%) where the SEPs were nor-mal and the TES-MEPs were abnormal.In only 1 case (2.6%) was the SEP normal and the MEP abnormal.The false negative rate was 0% with combined SEP and TES-MEP monitoring,while the false positive rate was 5.3%.There were 2 cases complicated by post-operative neurological deficits.(2) Spontaneous EMG monitoring can accu-rately determine the functioning of lumbar nerve roots during lumbar or lumbosacral tuberculosis surgery.Among 5 cases where EMG responses were observed,4 cases occurred during the spinal canal and nerve root decompression,1 case occurred in the orthopedic reset phase.Conclusions (1) During tuberculosis surgery involving thoracic,lumbar or sacral vertebrae,combined monitoring of SEPs and TES-MEPs can reflect the physiological and pathological condition of the spinal cord after ruling out interfering factors.This can improve monitoring and help assure the safety of lumbar surgery.(2) Intra-operative s-EMG monitoring can accurately reveal nerve root function in real time,help-ing to avert nerve root injury in lumbar and lumbosacral tuberculosis surgery.
9.Investigation of the carotid intima-media thickness in 221 individuals with metabolic syndrome
Wen-Sheng JIN ; Chang-Yu PAN ; Ju-Ming LU ; Guang ZHI ; Bo YANG ;
Chinese Journal of Endocrinology and Metabolism 1986;0(03):-
Metabolic abnormalities were identified and carotid intima-media-thickness(IMT)was measured in 221 individuals at risk for metabolic syndrome(MS).The results indicated that IMT was significantly thicker in MS individuals than that in non-MS individuals(P<0.01).And there was a tendency of progressive increase in IMT with increasing components of metabolic syndrome.