Objective To obtain the heavy chain variable region (VH) gene of monoclonal antibody 2F2 against Japanese encephalitis virus (JEV).Methods Total RNA was isolated with Trizol from hybridoma 2F2 cells,and cDNA of VH was amplified with reverse transcription polymerase chain reaction (RT-PCR) and sequenced.The putative VH gene was expressed in E.coli,and the expressed products was detected with enzyme linked immunosorbent assay (ELISA) to determine the activity to bind JEV.Results The VH gene was 354 bp in length which encodes 118 amino acids.Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) showed that the VH gene was successfully expressed and purified from inclusion bodies.ELISA result also demonstrated that VH gene expressed products bind purified JEV.Conclusions The VH gene of monoclonal antibody 2F2 against JEV had been cloned.

Objective To isolate and identify the putative Japanese encephalitis virus (JEV) re-ceptors from C6/36 and Vero cells. Methods Molecules binding with JEV were isolated from C6/36 and Vero cells by co-immunoprecipitation (Co-IP) approach, identified by mass spectrometry, and detected by Western blot. The location of putative JEV receptor on cells membrane and the binding with JEV were ob-served by laser scanning confocal microscopy (LCM). Results Several molecules binding with JEV were isolated from C6/36 and Vero cells by Co-IP, and only one molecule was identified as heat shock cognate 70 (HSC70) by mass spectrometry. Antibody against HSC'70 was able to detect a 74 ×103 protein isolated by Co-IP from C6/36 and Vero cells membrane in Western blot assays. It was observed by LSCM that when JEV attached on the surface of C6/36 cells, JEV and HSCT0 protein were co-localization. Conclusion 74 x 103 molecular identified as HSC70 protein from C6/36 cells may be JEV receptor.

Objective To screen and identify effective small interfering RNA (siRNAs) targeting the conserved 5’ untrans-lated region (UTR) of the enterovirus 71 (EV71) genome. Methods Double-stranded siRNAs were designed to target the 5’UTR of the EV71 genome. The cytotoxicity effect of siRNAs on rhabdomyosarcoma (RD) cells was evaluated. The cytopathic effect on EV71-infected RD cells was observed under phase-contrast microscopy and the effective siRNAs were screened out by cell viability assay and real-time TaqMan RT-PCR assay. Results All the siRNAs did not exhibit any cytotoxic effects on the viability and growth of RD cells. Transfection of si-1 and si-2 targeting the 5’ UTR in EV71 genome into RD cells signiifcantly delayed and alleviated the cytopathic effects of EV71 infection, increased cell viability and reduced the EV71 RNA transcripts. And the inhibitory effect of si-1/si-2 on EV71 replication was sequence-speciifc. Conclusions The 2 siRNAs (si-1 and si-2) targeting the conserved 5’UTR of the EV71 genome may have broad spectrum antiviral effects on many epidemic EV71 strains.

Objective:New Zealand rabbits were immunized with VLPs-MEpS,VLPs-E2S,and the levels of neutralizing antibodies in serum were determined.Methods: New Zealand rabbits were immunized with 10 μg VLPs-MEpS and VLPs-E2S,serum was collected at diffferent time with a two-weeks interval.The neutralizing antibodies were determined by ELISA.HCV(type 1b) had been prepared and mixed with serum from immunized rabbit before infected Huh7.5 cell.The protection of neutralizing antibodies in serum was assessed.Results: Neutralizing antibodies had been induced in rabbit after immunized with VLPs-MEpS and VLPs-E2S.VLPs-MEpS group had higher titer of antibodies than that of VLPs-E2S group(P<0.05),both group had higher titer of antibodies than that of control groups significantly(P<0.01).VLPs-MEpS group had higher neutralization than that of VLPs-E2S group(P<0.05),the highest neutralization rate was 61.49%.Both groups were higher than control group notably(P<0.01).Conclusion: Protective neutralizing antibodies have been induced in New Zealand rabbit after immunized with VLPs-MEpS and VLPs-E2S.It′s the basement for development of neutralizing antibodies vaccine.

Objective To establish WeChat official account-based platform for evidence dissemination,and to evaluate the effects of the platform.Methods The WeChat official account-based platform for evidence dissemination was established,and big data analysis and sampling survey were adopted to analyze information and its communication effects published from December,2014 to September,2016.Results Totally 22 369 followers used the platform and conducted 404 232 hits on its pages in total.The most frequent searches were Evidence and Knowledge of evidence-based nursing.The WeChat Communication Index was 433.07.The overall evaluation score was 4.34± 0.67.Conclusion WeChat official account-based platform for evidence dissemination can promote the accessibility of evidence and receives high evaluation score from followers.

Objective To evaluate the initial experience of intracoronary stenting in patients with acute myocardial infarction (AMI) in our hospital.Methods Ballon-expansion stents were deployed in 18 patients (male: 15, female 3, aged between 38-72 years old) with AMI after emergency PTCA. The major indications for intracoronary stenting in our present study include: 1) acute reocclusion or high risk of reocclusion due to intimal dissection; 2) severe residual stenosis (≥50% diameter stenosis) after repeat balloon dilation; and 3) obvious elastic recoil failed to response to repeat intracoronary nitroglycerin infusion. The dilating pressure for stent implantation was 12-18 atm with a dilating time of 10-30 sec. Patients were heparized during catheter maneuvers and were medicated with Ticlopidine (250 mg, twice daily) for 3 months and aspirin (250 mg, once daily) after stenting.Results 1) Coronary angiography (CAG) showed that all patients (n=18) had a single-vessel total occlusion (left anterior decending 9, right coronary artery 8, and left circuflex 1) before emergency PTCA and was successfully restored to TIMI 3 grade blood flow after intracoronary stenting (13 with Nir stent, 1 with Jonson and Jonson stent, and 4 with Cordis stent). 2) Minor residual intracoronary thrombosis was presented in 5 of 18 patients after PTCA, and it was totally disppeared after stenting; 3) One patient with inferior infarction developed Ⅲ degree atrioventricular blockade (AVB) and temporary pacemaker was introduced. This patient died of cardiac tamponade 6 h after stenting due to right ventricular perforation by electrode. No cardiac death, recurrent angina and reinfarction occurred during the 4-16 months follow-up peroid in the other 17 patients; 4) No angiographically restenois was found in all the 3 patients who had a secondary conronary angiography 4 weeks after stenting. 5) Left ventricular ejection fraction (LVEF) determined with Doppler echocardiography 4 weeks after stenting tended to be improved but failed to reach significant difference as compared to the basal LVEF (43.27%±8.43% vs 40.58%±7.23%, P>0.05, n=17) measured within 24 h after the onset of chest pain.Conclusions These results suggest that intracoronary stenting is a highly effective strategy in prevention or treatment of acute reocclusion after emergency PTCA in AMI. Minor residual intracoronary thrombosis after balloon dilation was not a contraindication for stent implantation.

OBJECTIVETo investigate the effect of carvacrol pretreatment on myocardial ischemia-reperfusion (I/R) injury and its underlying mechanisms.

METHODSWild-type male C57 BL/6 mice were randomized into 5 groups (n=13), namely the sham-operated group, vehicle (DMSO in saline)+ I/R group, carvacrol (20 mg/kg) + I/R group, carvacrol (40 mg/kg) + I/R group, and carvacrol (60 mg/kg) + I/R group. The mouse models of myocardial I/R injury were established by a 45-min occlusion of the left anterior descending coronary artery (LAD) followed by reperfusion for 2 h. Carvacrol or vehicle was administered intravenously 15 min before LAD occlusion. After reperfusion, the mice were examined for myocardial oxidative stress level and apoptosis rate.

RESULTSCompared with the vehicle group, the 3 carvacrol-pretreated groups showed significantly reduced myocardial infarct size, oxidative stress level and cardiac myocyte apoptosis rate (P<0.01).

CONCLUSIONCarvacrol can protect against myocardial I/R injury by inhibiting myocardial oxidative stress and apoptosis in mice.

Animals ; Antioxidants ; pharmacology ; Apoptosis ; drug effects ; Cardiotonic Agents ; pharmacology ; Male ; Mice ; Mice, Inbred C57BL ; Monoterpenes ; pharmacology ; Myocardial Infarction ; pathology ; Myocardial Reperfusion Injury ; metabolism ; pathology ; Myocytes, Cardiac ; cytology ; drug effects ; Oxidative Stress ; drug effects ; Random Allocation

In the early 1990's, emergency percutaneous transluminal coronary angioplasty (PTCA) has been proven to be the best strategy for acute myocardial infarction (AMI). It has not only resulted in a higher vascular recanalization, but has also further reduced mortality and morbidity, and has greatly improved life quality of patients with AMI.Objective To investigate the effect of emergency PTCA in AMI.Methods Emergency PTCA was performed in 52 AMI patients (male: 43, female: 9) from October 1993 to December 1997 in our hospital. The patients were aged between 31-89 years (±s, 53.7±8.2). Among them, 25 had anterior infarction, 27 inferior infarction. Angiographically infarct-related vessels were left anterior descending artery (LAD) in 27 cases, right coronary artery (RCA) in 22, and left circumflex artery (LCx) in 3. The mean time from the onset of chest pain was 6.12±5.13 h (2-12 h). Primary PTCA (balloon angioplasty without previous thrombolytic therapy) was performed in 34 cases, and rescue PTCA (balloon angioplasty after previous thrombolytic therapy that had failed to restore blood flow) in 15 cases, and immediate PTCA ( balloon angioplasty after previous successful thrombolysis but with severe residual stenosis and reoccurring angina pectoris) in 3 cases. Only the infarct-related vessels was dilated during emergency PTCA which is similarto conventional balloon angioplasty. All patients were given aspirin of 250 mg/d and were heparinized during the first 3-5 days after PTCA. Ticlopidine (250 mg, twice daily) was also given for 3 months if patients had experienced intracoronary stenting.Results Emergency PTCA was successfully performed in 48 of 52 patients (33/34 in primary PTCA, 12/15 in rescue PTCA, and 3/3 in immediate PTCA) which accounts for the initial success rate of 92%. Intracoronary stents were implanted in 18 of the 48 patients due to the presence of different vessel complications. PTCA was failed in 4 patients (3 occurred in rescue PTCA, 1 occurred in primary PTCA). Among the patients, one died of acute pulmonary edema during the rescue PTCA procedure, one had acute reocclusion after rescue PTCA and died of acute pump failure within 24 h. Guiding wire failed to put through lesions in the other 2 patients. Mass intracoronary thrombosis was present in 5 of 34 patients after primary PTCA despite the restoration of TIMI 3 grade blood flow in the distal segment. The residual thrombosis disappeared after urokinase (500 000-1 500 000 IU) was infused into the culprit coronary.Conclusions Clinical studies have suggested that emergency PTCA is more effective in early reperfusion than that of thrombolysis in AMI. Though emergency PTCA is highly effective, but the initial success rate is lower than that of conventional PTCA with a higher complication and mortality. Thus it should be only steadily introduced into AMI.

Objective To systematically retrieve,evaluate and integrate the best evidences on the early fluid resuscitation management in the patients with acute pancreatitis(AP)at home and abroad to provide ref-erence for clinical decision.Methods The related evidences on the early fluid resuscitation management in the AP patients were retrieved by computer from the databases of BMJ Best Practice,Up To Date,JBI,National Institute for Health and Care Excellence,Registered Nurses Association of Ontario,Guideline International Network,Scottish Intercollegiate Guidelines Network,International Association of Pancreatology,American Pancreatic Association,American College of Gastroenterology,Yimaitong,Cochrane Library,PubMed,Em-bass,CINAHL,The Web of Science,CNKI,Wanfang databases.The retrieval time limit was from the data-base establishment to March 20,2022.The literatures types included thematic evidence summarization,guide-lines,evidence summaries,systematic reviews and expert consensus.The researchers conducted the literature quality evaluation.The literatures meeting the standard conducted the evidence extraction.Results A total of 13 arti-cles were included,including 3 special subject evidence summary,4 guidelines,2 evidence summary,2 systematic evalu-ation and 2 expert consensus.A total of 16 pieces of best evidence were integrated,involving 4 aspects of organization management,evaluation and monitoring,fluid infusion strategy and health education.Conclusion It is recommended to use the target-oriented therapy for early fluid resuscitation management,and perform the fluid resuscitation immediate-ly after diagnosis,according to the patient's underlying disease,disease changes and monitoring indicators,implement precise early fluid resuscitation in order to reverse pancreatic microcirculation disorder,increase tissue perfusion and improve the patient's prognosis.

Objective To investigate the effect of carvacrol pretreatment on myocardial ischemia-reperfusion (I/R) injury and its underlying mechanisms. Methods Wild-type male C57 BL/6 mice were randomized into 5 groups (n=13), namely the sham-operated group, vehicle (DMSO in saline)+I/R group, carvacrol (20 mg/kg)+I/R group, carvacrol (40 mg/kg)+I/R group, and carvacrol (60 mg/kg)+I/R group. The mouse models of myocardial I/R injury were established by a 45-min occlusion of the left anterior descending coronary artery (LAD) followed by reperfusion for 2 h. Carvacrol or vehicle was administered intravenously 15 min before LAD occlusion. After reperfusion, the mice were examined for myocardial oxidative stress level and apoptosis rate. Results Compared with the vehicle group, the 3 carvacrol-pretreated groups showed significantly reduced myocardial infarct size, oxidative stress level and cardiac myocyte apoptosis rate (P<0.01). Conclusion Carvacrol can protect against myocardial I/R injury by inhibiting myocardial oxidative stress and apoptosis in mice.