Objective To investigate the molecular characteristic,the virulence factors and antimicrobial resistance of Methicillin-resistant Staphylococcus aureus (MRSA) isolated from pediatric patients.Methods Ninety-eight non-duplicate strains of and 49 non-duplicate strains of Methicillinsusceptible Staphylococcus aureus (MSSA) isolated from the three children's hospitals in Shanghai in 2008 were investigated.Panton-valentine leukocidin (PVL) gene was detected by polymerase chain reaction (PCR).The genotypes of staphylococcal cassette chromosome mec (SCCmec) of the MRSA isolates were confirmed by multiplex PCR.The sequence type (ST) of each strain was determined by multilocus sequence typing (MLST),and the algorithm eBURST was used to identify groups of clonal complex (CC).The minimal inhibitory concentrations (MIC) of fourteen antibiotics for all isolates were determined by agar dilution method.Results Among 98 isolates of MRSA,the positive rate of PVL genes was 6.1％ (6/98).In contrast,the positive rate of PVL genes was 4.1％ (2/48) of the MSSA strains.Among 98 isolates of MRSA,4.1％ (4/98),23.5％ (23/98),53.0％ (52/98) and 15.3％ (15/98) of the strains harboured SCCmec types Ⅱ,Ⅲ,Ⅳ and Ⅴ,respectively. The remaining four isolates (4.1 ％) presented a unique SCCmec pattern that could not be classified to any known types by the employed typing assays.Combining the ST and SCCmec type,the predominant clones were ST59-SCCmec Ⅳ (30 strains) and ST239-SCCmec Ⅲ (23 strains),followed by ST5-SCCmecⅣ and ST1-SCCmecⅣ (8 strains for each clone),ST239-SCCmec Ⅴ (6 strains),ST88-SCCmecⅤ (5 strains),ST5 SCCmecⅡ (4 strains),ST59-SCCmec Ⅴ (3 strains),ST8-SCCmecⅣ and ST88-SCCmecⅣ (2 strains for each clone),ST22-SCCmecⅣ,ST910-SCCmecⅣ and S45-SCCmec Ⅴ (1 strain for each clone),eBURST analysis distributed the MRSA isolates into several CC.ST8 and ST239 belonged to ST8 CC,ST1 belonged to ST15 CC,ST910 belonged to ST 30 CC,ST59,ST5,ST88,ST45,ST22,ST9 and ST7 were the origin of their own CC.The results of MIC showed that the 67 strains of MRSA harboring SCCmec type Ⅳ or SCCmec type Ⅴ were more susceptible to various non-β-lactam antibiotics than 27 strains of MRSA harboring SCCmec type Ⅱ or SCCmec type Ⅲ,and no vancomycin-resistant strain was found.Conclusions In three children's hospitals in Shanghai,the PVL gene-positive rate of MRSA isolates is relatively low,SCCmec type Ⅳ and SCCmec type Ⅴ could spread among hospitals to cause a small scale epidemic and have a variety of ST.

Objective To study the correlation between apolipoprotein E (APOE) genetic polymorphisms and sepsis in Chinese children.Methods The inpatients suffered with sepsis were enrolled as septic group and the healthy children from child health division were enrolled as control group.The study of APOE genotypes were carried out by polymerase chain reactions followed a high-resolution melting curve analysis.SPSS 16.0 statistical software was used for data analysis.Mann-Whitney U test was used to compare the age between the groups.Hardy-Weinberg equilibrium was tested using the Pearson x2-test.The x2-test was used to compare gender and the genotype distribution between the groups.The odd ratio (OR) was calculated together with its 95％ confidence interval (CI).Potential confounding effects of variables were corrected using a multivariate unconditional logistic regression model.All statistical tests were two-sided and P ＜ 0.05 indicates statistically significance.Results Among a total of 285 children collected from March 2011 to June 2012,there were 88 patients with sepsis and 197 healthy children.In the septic group,15 septic patients were complicated with central nervous system infection.Four apolipoprotein E genotypes were identified to be ε3/ε3,ε2/ε3,ε3/ε4,and ε2/ε4.The percentage of each genotype found in patients of the septic group and the control group was 64.4％ vs.73.1％ (ε3/ε3);16.8％ vs.10.7％ (ε2/ε3);18.8％ vs.14.7％ (ε3/ε4);0％ vs.1.5％ (ε2/ε4),respectively.The number of patients with the genotype ε3/ε3 among septic patients was significantly lower than that among the control individuals (P =0.047,1-β =0.334,OR =0.585,adjusted OR =0.559).The number of patients with the genotype ε3/ε3 among the septic patients with central nervous system infection was 33.3％,which was also significantly lower than that among the septic patients without CNS infection (67.1％).(P =0.014,1-β5 =0.685,OR =0.245,adjusted OR =0.275).Conclusions Apolipoprotein E genetic polymorphisms were associated with the occurrence of sepsis and central nervous system complications in children.The susceptibility of children with genotype ε3/ε3 to sepsis and central nerve system infection complications is significantly lower than that of children with other genotypes.

Objective To determine the drug-resistance rate of Enterococci isolated from patients of 5 padiatric hospitals located at different areas in China,and to investigate the distribution of resistance genes ermB,mefA,tetM and the integrase gene intTn of Tn1545 in Enterococci.Methods The antimicrobial susceptibility to 8 antibiotics of 2 216 Enteroeocei isolates was determined.PCR was used to detect the macrolide resistance genes ermB and mefA,tetracycline resistance genes tetM,and the integrase gene int-Tn of Tn1545.Results The resistance rates to erythromycin,ampicillin,gentamicin and teicoplanin were 86.5%,48.0%,60.5% and 0.7%,respectively.All isolated Enterococci straim were found sensitive to vancomycin.Of the detected 225 strains,70.7% of the 225 detected strains carried ermB gene while 75.1% of them carried tetracycline resistance gene tetM:only one strain had mefA.The presence of ermB gene in erythromycin MIC＞256 mg/L straim group(95.7%)strains was higher than those in erythromycin MIC＜256 mg/L group(2.5%).The int-Tn gene was detected in 40.9%(92/225)of the 225 test strains.The presence of ermB gene in int-Tn positive group strains was higher(84.8%)than those in int-Tn negative strains group(60.9%).So did the tetM in int-Tn positive group(83.7%)compared with those in int-Tn negative group(70.0%).Conclusions Enterococci sbowed a high resistance rate to the antibiotics we monitored,especially to erythromycin;but still very senstive to glycopeptide antibiotics. Resistance to macrolide in Enterococci collected from clinical in five Children's Hospital was generally mediated by methylation of 23S rRNA via ermB methylase. Enterococci resistance to tetracycline was predominantly due to ribosomal protection encoded by tetM. There was a strong relationship of the ermB and tetM genes with Tn1545-related elements.

Objective: To investigate the incidence and risk factors of catheter-related venous thrombosis (PICC-DVT) after peripherally inserted central catheter (PICC) in patients with hematologic malignancies, and to analyze the safety of anti-coagulation therapy with low-molecular-weight heparin. Methods: From August 2016 to June 2018, 43 patients with hematologic malignancies received PICC in Baoan District People's Hospital of Shenzhen City were enrolled. The patients were divided into low-molecular-weight heparin anticoagulation group (22 cases) and blank control group (21 cases) according to the random number table method. The blood routine, coagulation quadruple, D-dimer, protein C activity, protein S activity, and antithrombin Ⅲ activity before and after catheterization were compared between the two groups. Results: Of the 43 patients, 5 cases (11.62%) occurred PICC-DVT within 1 month after PICC, including 2 cases (9.09%) in the low-molecular-weight heparin anticoagulation group, and 3 cases (14.29%) in the blank control group, the difference between the two groups was not statistically significant (P = 0.664). No pulmonary embolism occurred in all patients with PICC-DVT. One case in the blank control group developed PICC-DVT and catheter-associated staphylococcus aureus infection, the patient was extubated after anti-infection and thrombolytic therapy, the other patients with PICC-DVT were not extubated, and the thrombus was dissolved after anticoagulant therapy. There were no significant differences in the white blood cell count, platelet count, prothrombin time, activated partial thromboplastin time, D-dimer, protein C activity, protein S activity, and antithrombin Ⅲ activity between the low-molecular-weight heparin anticoagulation group and blank control group (all P > 0.05). The anticoagulant index (protein C, protein S or antithrombin Ⅲ activity) was decreased in 5 patients with PICC-DVT, and in 38 non-thrombotic patients, the anticoagulant index was reduced in 16 patients (42.11%), the difference was statistically significant (P = 0.021). Conclusions The incidence of protein C, protein S or antithrombin Ⅲ activity reduction in hematological malignancies patients with PICC-DVT is higher than that in non-thrombotic patients. Low-molecular-weight heparin anticoagulant therapy can not reduce the occurrence of PICC-DVT within 1 month after PICC in patients with hematological malignancies, but the treatment is safe and has no relevant bleeding event.

Viral-encoded microRNAs (miRNAs) have vital roles in the regulation of virus replications and host immune responses. The results of previous studies have indicated that miRNA clusters are involved in the replication and virulence of the pseudorabies virus (PRV), which may potentially lead to immune escape or facilitation of PRV replication. This study's previous research revealed that prv-miR-LLT11a was differentially expressed during PRV infection. The present study's results have demonstrated that prv-miR-LLT11a could significantly inhibit PRV replication. It was further determined that SLA-1 was the target gene of prv-miR-LLT11a, and simultaneously, that overexpression of prv-miR-LLT11a could downregulate the mRNA and protein levels of SLA-1 in a dose-independent manner. Furthermore, the present study also observed that prv-miR-LLT11a can downregulate TAP1 expression. Our findings provide a better understanding of the molecular mechanism involved in the effects of prv-miR-LLT11a on SLA-1 and TAP1 as well as its involvement in immune system evasion of PRV.
Herpesvirus 1, Suid ; Immune System ; MicroRNAs ; Pseudorabies ; RNA, Messenger ; United Nations ; Virulence ; Virus Replication

Objective To explore the risk factors of incision complications after cerebral revasculariza-tion.Methods The patients who underwent vascular reconstruction for craniocerebral vascular diseases in the Affiliated Hospital of Xuzhou Medical University who underwent revascularization for cerebrovascular disea-ses from January 2018 to December 2022 were retrospectively included,and the patients were divided into the incision complications group and the no incision complications group.Among them,this study documented the delayed complications following incision healing after cerebral revascularization.The baseline clinical data of patients'demographics and incisional complications after cerebral revascularization were recorded,and possi-ble influencing factors were included in a binary logistic regression analysis to analyze the influencing factors that may contribute to the occurrence of incisional complications after cerebral revascularization.Results A total of 117 patients who underwent cerebral revascularization were included,including 25 cases(21.4％)in the group with incision complications,including 7 cases(28.0％)of fat liquefaction,4 cases(16.0％)of inci-sion dehiscence,7 cases(28.0％)of incision infection,and 7 cases(28.0％)of necrosis,while 92 cases(78.6％)in the group without incisional complications.Univariate analysis showed that there was no statisti-cally significant difference in gender,age,hypertension,hyperlipidemia,smoking,drinking,disease type,and vascular reconstruction type between the patients with incision complications after cerebral revascularization and those without incision complications(P＞0.05).The proportion of patients with diabetes mellitus and the frontal branches of superficial temporal artery not preserved in the incision complications group was higher than in the no incision complications group.BMI,scalp thickness,and operation time in the group with incision complications were higher than those in the no incision complications group,and the difference was statistical-ly significant(P＜0.05).Multifactorial analysis showed that scalp thickness(OR=1.484,95％CI:1.024-2.149,P=0.037),BMI(OR=1.323,95％CI:1.042-1.680,P=0.021),and the absence of the frontal branch of the superficial temporal artery(OR=3.582,95％CI:1.146-11.190,P=0.028),were the main risk factors for incisional complications(P＜0.05).Conclusion BMI,scalp thickness,and failure to preserve the frontal branch of the superficial temporal artery are independent risk factors for scalp incision complica-tions after cerebral revascularization.

OBJECTIVE: To detect variant of TRNT1 gene in a child featuring sideroblastic anemia with B-cell immunodeficiency, periodic fever and developmental delay (SIFD). METHODS: The proband and his parents were analyzed through trio-whole exome sequencing. Sanger sequencing and bioinformatic analysis were carried out to verify the candidate variant sites associated with the clinical phenotype. RESULTS: Genetic testing showed that the proband has carried compound heterozygous variants of the TRNT1 gene, namely c.88A>G(p.Met30Val) and c.363G>T(p.Glu121Asp). Sanger sequencing confirmed that the variants were respectively inherited from his father and mother. The variants were unreported previously. By bioinformatic analysis, both variants were predicted to affect the stability of binding of the TRNT1 protein with tRNA. Based on the American College of Medical Genetics and Genomics standards and guidelines, c.88A>G and c.363G>T variants of TRNT1 gene were predicted to be uncertain significance (PM2+PP3+PP4) and likely pathogenic (PM1+PM2+PP3+PP4), respectively. CONCLUSION The c.88A>G (p.Met30Val) and c.363G>T(p.Glu121Asp) compound heterozygous variants of the TRNT1 gene probably underlay the disease in this patient. Above finding has enriched the spectrum of TRNT1 gene variants.
Genetic Testing ; Humans ; Nucleotidyltransferases

Objective To investigate the resistance profile of clinical bacterial isolates to antibiotics in Shanghai during 2016.Methods Antimicobial susceptibility testing was carried out for the clinical isolates collected from 47 hospitals according to a unified protocol using Kirby-Bauer method or automated systems.The participating hospitals included 28 tertiary hospitals and 19 secondary hospitals across Shanghai.Results were analyzed according to CLSI 2016 breakpoints.Results A total of 122 548 clinical isolates were collected,including 35 522 (29.0％) strains of gram positive cocci and 87 026 (71.0％) strains of gram negative bacilli.Overall,28.9％ of the isolates were from secondary hospitals and 71.1％ from tertiary hospitals.Gram positive and gram negative isolates accounted for 25.8％ and 74.2％ in secondary hospitals,30.3％ and 69.7％ in tertiary hospitals,respectively.The overall prevalence of MRSA in Staphylococcus aureus was 48.7％ and 77.2％ of MRCNS in coagulase-negative Staphylococcus.The average prevalence of MRSA and MRCNS was 55.9％ and 73.3％ in secondary hospitals,45.9％ and 78.6％ in tertiary hospitals.No strains were found resistant to vancomycin in Staphylococcus.About 77.4％ of the 1 111 strains of non-meningitis S.pneumoniae isolated from children were penicillin-susceptible (PSSP),13.2％ were penicillin-intermediate (PISP) and 9.4％ were penicillinresistant (PRSP).The prevalence of PSSP,PISE and PRSP was 97.8％,2.2％,and 0 in secondary hospitals,76.5％,13.7％,and 9.8％ in tertiary hospitals.Of the 285 strains isolated from adults,94.0％,4.2％ and 1.8％ were PSSP,P1SP and PRSP,respectively.The prevalence of PSSP,PISP and PRSP among the isolates from adults was 93.7％,5.3％,and 1.0％ in secondary hospitals,94.2％,3.7％,and 2.1％ in tertiary hospitals.Overall,37 strains of vacomycin-resistant E.feacium (14 from secondary hospitals and 23 from tertiary hospitals) and 25 strains of vacomycin-resistant E.feacalis (all from tertiary hospitals) were identified.PCR and sequencing analysis indicated that most of these resistant strains were vanA type.The overall prevalence of ESBLs-producing srains was 52.2％ in E.coli,30.9％ in Klebsiella pneumoniae and 29.8％ in Proteus mirabilis.Specifically,the corresponding prevalence of such strains was 55.1％,33.6％ and 34.0％ in secondary hospitals,51.0％,29.7％ and 28.0％ in tertiary hospitals,respectively.Enterobacteriaceae strains were still highly susceptible to carbapenem antibiotics.Overall,8.9％ and 9.1％ of the Enterobacteriaceae strains were resistant to imipenem and meropenem,respectively.The figure was 6.6％ and 7.1％ in secondary hospitals,9.9％ and 10.0％ in tertiary hospitals.Extensively drug-resistant strains were identified in A.baumannii,K.pneumoniae,P.aeruginosa,and E.coli,specifically,223,63,10,and 4 strains in secondary hospitals;224,201,22,and 9 strains in tertiary hospitals.Conclusions Antibicotic resistance is still very serious in the common clinical strains,which poses a critical threat to healthcare facilities.This issue should be taken seriously and effective infection control measures must be put in place.

Objective To observe any curative effect of applying comprehensive pulmonary rehabilitation in moderate and severe cases of chronic obstructive pulmonary disease (COPD).Methods A total of 135 persons hospitalized with moderate or severe COPD were randomly divided into a rehabilitation group of 75 and acontrol group of 60.Both groups were given routine treatment,while the rehabilitation group was additionally provided with a comprehensive pulmonary rehabilitation regimen,including health education,exercise training,respiratory function training,respiratory muscle training,psychological support and nutritional intervention for six months.Before and after the treatment,both groups were evaluated using their walking distance within 6 minutes (6MWD),an anhelation index,a COPD assessment test (CAT),the Beck anxiety and depression scale,a nutritional assessment and indexes of pulmonary function and blood gases.Results After the intervention the average 6 MWD,anhelation index,CAT score,Beck anxiety and depression scores,forced expiratory volume,forced vital capacity and PaO2 of the rehabilitation group were all significantly better than before the treatment and better than those of the control group.Conclusion For moderate and severe COPD patients,comprehensive pulmonary rehabilitation effectively strengthens their moving ability,pulmonary function and arterial partial pressure of oxygen,while relieving anhelation,anxiety and depression.

Objective To investigate the antimicrobial resistance in the A cinetobacter baumannii strains in different parts of China during 2012 .Methods A total of 8 739 clinical isolates of Acinetobacter were collected from 13 general hospitals and two children’s hospitals ,of which most were A . baumannii (89 .6% , 7 827/8 739 ) . Antimicrobial susceptibility testing was carried out by means of Kirby-Bauer method according to the unified protocol . The susceptibility testing data were analyzed by WHONET 5 .6 software according to CLSI 2013 breakpoints .Results Majority (85 .4% ) of the Acinetobacter strains were isolated from inpatients .The remaining 14 .6% were from outpatients and emergency room patients .Of the 7 827 strains of A .baumannii , 10 .9% ,35 .2% ,35 .7% and 43 .4% were resistant to tigecycline ,minocycline ,cefoperazone-sulbactam and amikacin , respectively .The percentage of A .baumannii resistant to imipenem and meropenem was 63 .5% and 68 .2% ,respectively . The antimicrobial resistant pattern varied in different hospitals . The resistance of A . baumannii varied between different clinical departments .A number of pandrug resistant (PDR) (20 .0% ,1 567/7 827) and multidrug-resistant (MDR) (45 .0% , 3 521/7 827 ) A . baumannii were identified . Conclusions A . baumannii is the most popular pathogenic bacteria among Acinetobacter .The antibiotic resistance of A .baumannii is still increasing .Cefoperazone-sulbactam and minocycline has good in vitro antibacterial activity against A .baumannii .The antibiotic resistance of A .baumannii varies greatly with hospital and department .