Objective To investigate the prevalence and transmission mechanisms of plasmid-mediated blaoxa-23 resistance genes in Acinetobacter baumannii.Methods One hundred and one Acinetobacter baumannii were collected from Obstetrics and Gynecology Hospital of Fudan University and Renji Hospital Shanghai Jiaotong University School of Medicine.Antibiotic susceptibility of carbapenems were determined by standard agar dilution method.Molecular typing of Carbapenem-resistant Acinetobacter baumannii (CRAB) was performed by MLST.blaoxa-23、blaoxa-24、blaoxa-51、blaoxa-58、blaIMP-1、blaVIM-1/2 and blaAmp-C were analyzed by PCR.The analysis of blaoxa-23 transposons for carbapenems resistant A.baumannii isolates was also performed by PCR.Plasmid was analyzed by gel electrophoresis.Conjugation experiments were performed to determine the transferability of blaoxa-23.Results The antibiotic susceptibility tests showed the resistant rates to carbapenems were extremely high , and the ones of imipenem and meropenem were 64.4%and 69.3%.Fifty-six (53%) isolates were carbapenems-resistant A.baumannii.Main clone ST208 includes 28 isolates(50%) in CRAB.A total of the CRAB isolates harbored blaoxa-23 and blaoxa-51 (100%), 44 for blaIMP-1(78.6%) and 54 for blaAmp-C(96.4%), while blaoxa-24, blaoxa-58 and blaVIM-1/2 was undetected.Two previously identified transposons ( Tn2006 and Tn2008 ) was found in the isolates.Plasmid gel electrophoresis results showed that the isolates carried 2-4 plasmids and blaoxa-23 were transferable by plasmids.Conclusions There is high carbapenems resistance of A.baumannii infections.ST208 was the most prevalent molecular type.The mainly drug-resistant genes of A.baumannii are blaoxa-23.Based on the findings, blaoxa-23 is plasmid mediated, suggesting that it may transfer by plasmids carrying Tn 2008 transposon, thus induced isolates resistant to carbapenemase.

Objective To analyze the evolution and genetic characteristics of Echovirus 11 (Echo11) strains isolated from patients with hand food and mouth disease ( HFMD) in Henan province. Methods Enterovirus strains were isolated from stool samples of patients with HFMD from year 2010 to 2012.The sequences of VP1 region of all positive strains were amplified , sequenced and then analyzed by BLSAT to determine their genotypes .The entire VP1 coding regions of 10 Echo11 strains were amplified by RT-PCR, and the homology analysis was conducted among them and other Echo 11 strains published in Gen-Bank.A phylogenetic tree was constructed to evaluate the evolution of Henan isolates and the prevalence of different genotypes .Results From year 2010 to 2012 , 184 non-Enterovirus 71 ( EV71 ) strains and non-Coxsakievirus A16 (CA16) strains causing HFMD were isolated, 10 (5.43%) of with were Echo11 strains. There were 876 nucleotides ( nt) in the complete VP1 gene sequence , encoding 292 amino acids ( aa) .The 10 Echo11 strains shared 93.1%-100% homologies in nt sequences and 97.3%-100% in aa sequences. The homology analysis indicated that Henan strains and prototype strains were highly similar but different with the homology of 77.8%-78.8% in nt and 90.8%-91.8% in aa, respectively.Conclusion Echo11 was one of the pathogens causing HFMD in He-nan province and genotype A was the dominant genotype .

Objeetive To explore the correlative factors of early progressive motor deficits in cerebral infarction of moderately size in basal ganglia and the relationship between motor deficit progression and the stem lesion of middle cerebral artery.Methods One hundred and fifty patients with single acute infarction located in white matter of basal ganglia region were recruited in this study retrospectively.All patients were performed brain and neck MRI and magnetic resonance angiography (MRA) within 72 h after onset.They were divided into two groups based on the maximal diameter of infarction on diffuse weighing imaging(DWI):moderate size infarction group(75 patients,maximal diameter of infarction from 1.5 cm to 3.0 cm) and lacunar infarction group(75 patients,maximal diameter of infarction lower than 1.5 cm).The scores of dynamic National Institutes of Health Stroke Scale(NIHSS) in 7 d after the admission and multiple Logistic regression analysis were used and the risk factors of the early progressive motor deficits were analyzed.Results The rate of early progressive motor deficits in moderate size infarction group was significant higher than that in lacunar infarction group [32.0 ％ (24/75) vs.8.0 ％ (6f75)] (P ＜ 0.05).The multiple Logistic regression analysis showed that elevated systolic blood pressure on admission was the independent risk factor for early progressive motor deficits (P =0.016).The rate of stem lesion of middle cerebral artery in moderate size infarction group was significantly higher than that in lacunar infarction group [41.3 ％ (31/75) vs.9.3 ％ (7/75)] (P ＜ 0.01).Conclusions Patients with acute single infarction located in white matter of basal ganglia and with the diameter of 1.5-3.0 cm are more prone to early progressive motor deficits and elevated systolic blood pressure on admission is the most significantly independent factor.The pathogenic mechanism may be associated with the stem lesion of middle cerebral artery.

Objective To investigate the relationship between white-matter hyperintensities (WMHs) and hemorrhagic transformation and neurological outcome at 3 months after recombinant tissue plasminogen activator (rt-PA) treatment in patients with acute ischemic stroke.Methods The chnical data of 132 patients with acute ischemic stroke who had received intravenous rt-PA therapy were retrospectively reviewed.The severity of WMHs according to the modified Schelten scale was assessed.Hemorrhagic transformation included hemorrhagic infarct (HI) and cerebral parenchymal hemorrhage (PH).Hemorrhagic transformation after thrombolytic therapy and clinical neurological outcome based on modified Rankin scale (mRS) at 3 months was also analyzed.Favorable neurological outcome as mRS 0-1 score and unfavorable neurological outcome as mRS 2-6 scores was defined.Results Hemorrhagic transformation was found in 26 patients (19.7％,26/132) among 132 patients receiving intravenous rt-PA treatment.Seventeen patients were HI type and 9 patients were PH type.Multiple Logistic regression analysis showed that baseline American National Institute of Health Stroke Scale (NIHSS) score,diastolic pressure,WMHs score was not correlated with HI type hemorrhagic transformation (P ＞ 0.05) ; and WMHs score was not correlated with PH type hemorrhagic transformation (P ＞0.05),but baseline NIHSS score,diastolic pressure was the independent risk factor of PH type hemorrhagic transformation (P ＜ 0.05).Dicho-Logistic regression analysis showed that WMHs score was the independent risk factor of unfavorable neurological outcome (OR =1.136,95％ CI 1.037-1.245,P =0.008).Conclusion Severe WMHs are not associated with hemorrhagic transformation but independently associated with unfavorable neurological outcome after thrombolytic therapy in patients with acute ischemic stroke.

To explore the relationship between meteorological factors and incidences of fever, thrombocytopenia and leukopenia syndrome (FTLS), incidence data of FTLS in Xinyang City were collected and described. The relationship between FTLS and monthly meteorological factors such as average atmospheric pressure, average temperature, average relative humidity, average wind speed and precipitation was analyzed with simple correlation and multi-stepwise linear regression. The results showed that there was a significant negative correlation between incidence of FTLS and monthly average atmospheric pressure (P<0.01), and there were significant positive correlations between monthly average temperature, monthly average relative humidity, monthly precipitation and incidence of FTLS (P<0.05). The multi-stepwise regression showed that the regression equation was Y (monthly cases) =-12.70 + 0.28X2 (monthly average temperature), and the coefficient of determination was 0.76. The incidences of FTLS are correlated with temperature.

Objective:To analyze the inflammatory mechanism and potential intervention drugs related to angiotensin converting enzyme 2 (ACE2) inhibitory mutations in order to provide reference for the treatment of corona virus disease 2019 (COVID-19).Methods:The data of lung adenocarcinoma with ACE2 mutations were screened from the cancer genome atlas (TCGA) database. The data were analyzed by R program language edgeR package and cluster Profiler package, gene ontology (GO)functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis. Using String online analysis website for protein-protein interaction (PPI) network analysis, screening out the core genes, and finally using the Epigenomic Precision Medicine Prediction Platform (EpiMed) for multi-group association analysis of key genes, and drug candidates prediction.Results:A total of 1 005 differential genes were obtained, of which 91 were up-regulated and 914 down-regulated. A total of 71 GO were enriched, including 45 items related to biological processes, 16 items related to cell components, and 10 items related to molecular function. A total of 13 KEGG pathways were enriched, mainly in inflammatory pathways, various viral infectious diseases, transcriptional regulation, drug metabolism and protein digestion and absorption pathways. The differentially expressed genes were introduced into String online analysis website for PPI network analysis, a total of 252 proteins were obtained, and 10 core genes were H2A clustered histone 16(HIST1H2AL), H3 clustered histone 2 (HIST1H3B), H3 clustered histone 7 (HIST1H3F), H3 clustered histone 11 (HIST1H3I), H3 clustered histone 3 (HIST1H3C), H2B clustered histone 3 (HIST1H2BB), H2B clustered histone 6 (HIST1H2BI), H4 clustered histone 2 (HIST1H4B), H1-4 linker histone (HIST1H1E), H2A clustered histone 4 (HIST1H2AB). Interferon-α, resveratrol, celecoxib, heartleaf houttuynia herb, weeping forsythia capsule, dexamethasone, Chinese pulsatilla root, tumor necrosis factor-α inhibitors, liquorice root and famciclovir might be drugs for the treatment of ACE2 mutation-related inflammation.Conclusions:Inflammation associated with ACE2 inhibitory mutations is similar to the pathogenesis of COVID-19, which could lead to disease by promoting the activation of inflammatory pathways such as mitogen-activated protein kinase (MAPK), the Janus kinase signal transducer and activator of transcription (JAK/STAT), mammalian target of rapamycin (mTOR). Celecoxib, interferon and resveratrol may have the potential therapeutic effects on COVID-19.

OBJECTIVETo clone and express the recombinant capsid protein VP2 of enterovirus type 71 (EV71) and to identify the immune activity of expressed protein in order to build a basis for the investigation work of vaccine and diagnostic antigen.

METHODSVP2 gene of EV71 was amplified by PCR, and then was cut by restriction enzyme and inserted into expression vector pMAL-c2X. The positive recombinants were transferred into E.coli TB1, the genetically engineered bacteria including pMAL-c2X-VP2 plasmids were induced by isopropyl thiogalactoside ( IPTG) , and the expression products were analyzed by SDS-PAGE and western blotting method. EV71 IgM antibody detection method by ELISA was set up, and the sensitivity and specificity of this method was assessed; 60 neutralizing antibody positive serum samples from hand foot and mouth disease (HFMD) patients were determined, of which 52 samples were positive and 8 samples were negative; a total of 88 acute phase serum samples of HFMD patients diagnosed in clinical were also detected.

RESULTSVP2 gene of 762 bp was obtained by PCR, the gene segment inserted into the recombinant vector was identified using restriction enzyme digestion. The recombinant vector could express a specific about 71 500 fusion protein in E.coli by SDS-PAGE. The purified recombinant protein of EV71-VP2 can react with the serum of HFMD patients to produce a specific band by western blotting. The sensitivity and specificity of ELISA was 87% and 83%, respectively. Of the 88 acute phase serum samples from children with HFMD, 48 samples (55%) were positive by the ELISA assay.

CONCLUSIONSVP2 gene of EV71 has been cloned and a prokaryotic high expression system for VP2 gene was successfully constructed in the present study. The recombination EV71-VP2 has well antigenicity, which could be useful for developing diagnose reagent or vaccine of EV71.

Antibodies, Neutralizing ; blood ; Antibodies, Viral ; blood ; Capsid Proteins ; genetics ; immunology ; Enterovirus A, Human ; genetics ; immunology ; isolation & purification ; Enzyme-Linked Immunosorbent Assay ; Genetic Vectors ; Hand, Foot and Mouth Disease ; immunology ; Humans ; Immunoglobulin M ; blood ; Recombinant Proteins ; genetics ; immunology

The elbow is more susceptible to motion loss than other joints after trauma,and elbow stiffness leads to functional impairment in the upper limb and interferes with daily activities.Open arthrolysis is the most common and classical treatment for post-traumatic elbow stiffness.In this paper,we review the treatment protocols like preoperative clinical evaluation,arthrolysis strategies and postoperative rehabilitation program for post-traumatic elbow stiffness,discuss relevant issues and assess their prospects.

Objective:To investigate the omics mechanism of SARS-related immune injury and predict targeted therapeutic drugs through clinical bioinformatics analysis of the transcriptome data of SARS virus in order to provide reference for clinical treatment of COVID-19.Methods:The transcriptome data of SARA virus were collected from the Gene Expression Oibus (GEO) and used to screen differential genes. Enrichment analysis and protein interaction analysis were performed to investigate the mechanism of immune damage associated with SARS. A platform of epigenetics in precision medicine (EpiMed) was established to predict potential therapeutic drugs.Results:The mechanism of SARS-related immune injury was complex, involving affecting the function of immune cells through signaling pathways such as Toll-like receptors, increasing cytokines in plasma through Th17 signaling pathway and inducing autoimmune responses after autoantibodies were generated by molecules such as IL-6, NF-κB, and TNF. Drugs such as Chuanqiong and Etanercept might have therapeutic effects on SARS-related immune damage.Conclusions:SARS virus could cause abnormal expression of many immune-related molecules and signaling pathways. Drugs such as Chuanqiong and Etanercept might have therapeutic effects on SARS-related immune damage. This study might provide reference for clinical treatment of COVID-19.