AIM:To investigate the protective effect of hyperoside (Hyp) on Mycoplasma pneumoniae (MP) pneumonia (MPP) mice and the underlying regulatory mechanism.METHODS:BALB/c mice (n=40) were randomly divided into control group, model group, low-dose (12.5 mg/kg) Hyp group and high-dose (50 mg/kg) Hyp group.The MPP model was established using MP standard strains (MPFH) through nasal dripping.The treatment started on the second day, and the follow-up experiments were performed after 3 d.The lung histopathology were detected by HE staining.The levels of C-reactive protein (CRP), alanine aminotransferase (ALT), aspartate transaminase (AST), lactate dehydrogenase (LDH) and MB isoenzyme of creatine kinase (CK-MB) in the serum, and the levels of reactive oxygen species (ROS), superoxide dismutase (SOD) and glutathione (GSH) in the lung tissues were measured by automatic biochemical analyzer.ELISA and Western blot were used to detect interleukine-1 beta (IL-1β), IL-6, IL-18 and tumor necrosis factor-alpha (TNF-α) levels in the serum and lung tissues, respectively.The expression of NLRP3, ASC and caspase-1 at mRNA and protein levels was evaluated by RT-qPCR and Western blot.RESULTS:Compared with control group, the inflammation and alveolar interstitial widening were observed in the lung tissues of model mice, and the CRP, ALT, AST, LDH and CK-MB levels were significantly up-regulated (P<0.05).The ROS level was increased, while SOD and GSH levels were significantly decreased (P<0.05).The levels of IL-1β, IL-6, IL-18 and TNF-α were markedly up-regulated (P<0.05).The protein expression of NLRP3, ASC and caspase-1 was also obviously increased (P<0.05) in model group.Compared with model group, the symptoms above were significantly relieved in Hyp group, and the effect of high-dose Hyp was more apparent than that of the low-dose one.CONCLUSION:Hyperoside suppresses the injury in MPP mice, which may be related to the inhibition of oxidative stress and NLRP3 inflammasome activation.

Objective To study the education of medical mycology for undergraduates of medical laboratory specialty and provide a basis for teaching reformation.Method Setting of mycology related courses of medical mycology for undergraduates in 5 medical schools and 85 inspection and technical personnel's detection of fungi in 81 hospitals were investigated through consultation and questionnaire survey.Results More than 140 class hours for medical mycology were arranged in 5 schools,but as to medical mycology,22 class hours in 1 school and less than 10 class hours in 4 schools,the minimum class hours were 5.Although various numbers of Candida and filamentous fungi could be isolated in hospitals investigated,more than half laboratory workers could not identify penicillium,thermally dimorphic fungi,Zygomycetes and Dematiaceous fungi.Conclusion Education on medical mycology for medical laboratory specialty undergraduates is insufficient and the corresponding teaching lacks such content as medically important pathogenic fungi detection methods and identification characteristics.The hospital technical personnel's fungal identification ability cannot meet the situation of increasing fungal infection involved in clinical medicine,so it is necessary to carry out teaching reformation of medical mycology for undergraduates in laboratory medicine,including adding class hours,increasing course contents and so on.

ObjectiveTo establish the rapid molecular diagnosis of 16 common coagulase negative Staphylococcus(CNS).MethodsDNA sequencing of 16 CNS would be obtained with gap gene.After the alignment gap gene sequences which were available in the GenBank,the bacteria were identified with homological alignment and phylogenetic tree,and compared with the 16S rRNA gene.ResultsThe sequence similarity of the gap sequences ranged from 39% to 98% in 16 CNS.There were the highest similarity (98%) between S.hominis and S.hominis subsp,and the lowest(39% ) between S.saprophyticus and S.xylosus.The sequence similarity of the 16S rRNA sequences ranged from 96 to 98%,at least two species of bacteria similar rate of 99% and the most four species similar rate of 99%.Phylogenetic homology analysis showed that it was a high confidence(99% ) in the detection ofS.xylosus and S.lentus,S.chromogenes and S.intermedius,S.hominis and S.hominis subsp,but for 10 other species of bacteria,gap homology analysis has less unreliable confidence(49%,56% ) and 16S rRNA has more unreliable confidence(43%,43%,50%,56%,63%,65%,76% ).ConclusionAnalysis of gap sequence could identify 16 CNS timely and accurately,with higher confidence than 16S rRNA.

Objectives To investigate the immune effect of hepatitis B vaccine under the influence of congenital human cytomegalovirus (HCMV) infection. Methods The newborn rat model of congenital HCMV infection was developed by intra-peritoneally inoculating pregnant rat with HCMV suspension,while the offsprings of healthy rats were used as the control group. Offspring rats in all groups were inoculated with hepatitis B vaccine in the postnatal 1st, 3rd and 5th week and were taken blood from hearts separately in 3rd, 5th, 7th and 11th week. Antibody to Hepatitis B surface antigen (HBsAb) titer in all groups was de-tected by ELISA method. Results The serum HBsAb titer in both groups all showed a trend of increasing gradually with added vaccinating times and decreased differently with time extending after completed vaccinations. Differences among changes of HBsAb titer along with prolonged time in each group were all statistically significant (P<0.001). At all time points (3rd, 5th, 7th, 11th week), the titer of serum HBsAb in congenital HCMV infection group was lower than that in the control group respectively, and there were statistically significant differences (P<0.01). Conclusions Congenital HCMV infection could weaken the im-mune effect of hepatitis B vaccine.

Objective To detect the main virulence genes and biofilm formation of Enterococci strains isolated from blood samples .Methods Twenty-eight strains of Enterococcus faecalis ( E.faecalis) and 54 strains of Enterococcus faecium ( E.faecium) were collected from blood samples .Five main virulence genes (asa1, esp, hyl, cylA and gelE) were detected by multiplex PCR.Biofilm formation was investigated by using microtiter dish biofilm formation assay .Results All E.faecalis strains were positive for at least one kind of virulence genes , of which 14 strains were concurrently positive for asa1, esp, cylA and gelE.asa1, cylA and gelE were only detected in E.faecalis strains, while hyl gene only existed in E.faecium strains. Twenty-seven strains of E.faecium were esp positive, of which 12 strains were both hyl and esp positive. None of the 5 virulence genes were identified in 10 strains of E.faecium.85.7% of E.faecalis strains and 63.0%of E.faecium strains could form biofilm.Conclusion Compared with E.faecium strains, more types of virulence genes were detected in E.faecalis strains with higher positive rates .Moreover , E.faecalis strains were more likely to form biofilms than E.faecium strains.

Objective To detect the hypermucoviscosity phenotype , capsular serotype and virulence gene of Klebsiella pneumonia (K.pneumonia) from various kinds of clinical specimens and understand the characteristics of different K.pneumonia causing infections.Methods A retrospective study was conducted through collection of 178 K.pneumonia isolates from blood, sputum, bronchoalveolar lavage fluid , urine, normally sterilized fluid , puncture fluid from liver abscess and other abscesses between January 2010 and December 2012 in General Hospital of Chinese PLA.String test was carried out for detection of hypermucoviscosity phenotype.Capsular serotype and virulence gene ( rmpA) were checked by polymerase chain reaction.Analysis was made according to the hypermucoviscosity , capsular serotype , rmpA gene , as well as the sources of K.pneumoniae.Statistic data was analyzed by contingency table analysis and χ2 test.Results Eighty-three out of 178 ( 46.6%) strains of K.pneumonia were hypermucoviscous with positive string test, the positive rate of virulence gene rmpA was 92.8%(77/83).K1/K2/K57 capsular serotypes were the predominant serotypes in the group of puncture fluid from liver abscess and other abscesses (75.0%,27/36)than the group of blood (32.4%,12/37), urine(21.7%,5/23) and normally sterilized fluid(25.0%,5/20), and also more than in the group of sputum , bronchoalveolar lavage fluid (50.0%,22/44),χ2 =21.19,P<0.01.The positive rate of string test in the group of puncture fluid from liver abscess and other abscesses (77.8%, 28/36)was significantly higher than the group of blood (29.7%, 11/37), urine (30.4%, 7/23), or normally sterilized fluid (25.0%, 5/20),χ2 =27.90,P<0.01.The positive rate of rmpA gene in the isolates from puncture fluid of liver abscess and other abscesses was higher than other groups.Conclusions As the pathogens of various kinds of infections , mainly abscess and respiratory infection, hypermucoviscous strains of K.pneumonia were of great clinical significance.Capsular serotype K57, as well as K1 and K2, possessed hypermucoviscosity and hypervirulence in China.

A rapid and sensitive method for the determination of trace carbaryl in water by using diallyl phthalate-europium ( Eu3+) as fluorescent probes was developed. The interaction between Eu3+ and diallyl phthalate and carbaryl was studied by high resolution mass spectrum, and the fluorescence spectra change of complexes before/after binding with carbaryl was also investigated. The influence factors of fluorescence intensity including solution pH and interferent were studied. The results showed that two diallyl phthalate molecules were complexed with one Eu3+to form stable complexes. Carbaryl could also interact with the probe to form multiple complexes, which significantly increased the fluorescent efficiency of the probe. At pH 9. 0 of solution and by using 245/615 nm as excitation/emission wavelength, the fluorescence intensity showed good linear relationship with the carbaryl concentration ranged from 6. 25í10-8 mol/L to 2. 50í10-6 mol/L, and the linear correlation coefficient was 0. 9968. The detection limit of the method was 9. 6í10-9 mol/L. Water samples were extracted by acetonitrile, and then detected by europium ( Eu3+)-diallyl phthalate fluorescent probe. The recovery of the method was 91. 8%-94. 5%, while RSD was within 6. 1%. The method is suitable for the rapid determination of carbaryl in water samples.

A method for rapid determination of semicarbazide in water by hydrophilic interaction chromatography-quadrupole / electrostatic field orbitrap high resolution mass spectrometry was developed. The sample was extracted with acetonitrile after 0. 1 mol/ L NaOH was added in the sample and then excessive amounts of Na2 SO4 was added to stratify acetonitrile from the mix solution. The acetonitrile extraction solution was dehydrated with anhydrous sodium sulfate. The preparation was separated by an amide column using as hydrophilic interaction column, and gradient elution program was employed by using water and acetonitrile containing 0. 1% formic acid as mobile phase, then it was detected in positive and selected ion monitoring mode by a quadrupole / electrostatic field orbitrap high resolution mass spectrometry. Internal standard method was used for quantitative analysis. The linear correlation coefficient of semicarbazide was 0. 997 in the concentration range of 0. 2 -20 μg / L under the optimal conditions. The limit of detection was 0. 09 μg / L, while the limit of quantitation was 0. 30 μg / L. The recoveries were 82. 3% to 92. 0% , and the relatively standard deviations were less than 7. 6% at the spiked levels of 0. 5, 1. 0 and 5. 0 μg / L using river water and sea water as blank samples. The developed method is suitable for the analysis of trace semicarbazide in environment water samples.

Objective To investigate the influence of infection factors on kidney transplantation after organ donation and possible countermeasures.Methods Thirty-seven cases of kidney transplantation in Organ Transplantation Center of Qianfoshan Hospital Affiliated to Shandong University from January 2014 to December 2016 were retrospectively analyzed.The patients were divided into two groups according to perioperative infection prevention programs:42 patients with postoperative routine use of cephalosporins or penicillin for 2 weeks,and 95 patients with postoperative application of carbapenems + micafungin.Postoperative infection rate,occurrence time,pathogen infection;donor age,perioperative pathogens of donor and receptor (organ preservation solution,drainage fluid,urine,sputum samples),acute rejection,delayed graft function (DGF),diabetes mellitus,and the use of immunosuppressive agents were recorded.Results The infection rate in carbapenem + micafungin group was 12.6％,and the infection rate in cephalosporin or penicillin group was 19.4％ (P＜0.05).Pathogen positive detection rate of the drainage fluid,urine and sputum was lower in carbapenems + micafungin group than that in cephalosporins or penicillin (P＜0.05).Within 2 weeks after operation,the detection rate of bacteria and fungi in the carbapenems +micafungin infection prevention group was lower than that in the control group (P＜0.05).There was no significant difference in the detection of viruses (P＞0.05).There were no significant differences in the detection of pathogens among the two weeks to six months after surgery (P＞ 0.05).Donor infection,acute rejection,DGF,and diabetes mellitus were the risk factors for postoperative infection (P＜0.05).Conclusion The application of carbapenems and micafungin can reduce the incidence of infection for the early stage of DCD kidney transplantation.Donor infection,acute rejection,DGF and diabetes mellitus are all risk factors for the postoperative infection.

Objective:To investigate the resistance and transmission mechanism of carbapenem-resistant Enterobacterales (CRE), so as to provide the scientific evidence for the treatment and prevention of CRE infection.Methods:Seventy-six isolates of CRE isolated from Shaoxing Second Hospital between May 2016 and August 2018 were included. The isolates were re-identified by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS). The minimum inhibitory concentrations (MICs) of colistin, tigecycline, ceftazidime-avibactam, fosfomycin and other antibacterial drugs were determined using broth microdilution or agar dilution methods. PCR and sequencing analysis were performed to detect carbapenemase encoding genes ( blaKPC, blaNDM, blaIMP, blaVIM, blaOXA-48). Pulsed field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) were used to analyze homology of strains. S1-PFGE combined with Southern blot hybridization were used to locate the carbapenamase genes. Filter mating test were performed to determine the horizontal transfer ability of plasmids harboring carbapenamase genes. Results:Among the 76 isolates of CRE, 51 isolates were Klebsiella pneumoniae; 10 isolates were Escherichia coli; 15 isolates were other Enterobacterales. The 76 CREs were mainly isolated from urine, sputum and blood samples. The distribution rate of ICU was the highest (55.26%). The 76 CREs showed low resistance rates (0%, 1.33%, 18.42%) to colistin, tigecycline and ceftazidime-avibactam. The resistance rates to amikacin and fosfomycin were <45%, and the resistance rates to other drugs were >97%. The detection rate of KPC-2 carbapenemase was the highest (85.33%). The ST11 CRKP producing KPC-2 carbapenemase accounted for the highest proportion (62.75%), mainly distributed in the ICU (62.50%). Southern blot hybridization showed that blaKPC-2 was mainly located on a plasmid about 90 kb (39/63). Filter mating test showed that blaKPC, blaNDM and blaIMP could be transferred horizontally to recipient bacteria through plasmids. Conclusions:The 76 CRE isolates were only susceptible to a few antibacterial drugs, such as colistin, tigecycline and ceftazidime-avibactam. The production of KPC-2 carbapenemase was the main reason for the resistance of Enterobacterales to carbapenems. KPC-2 carbapenemase-producing ST11 Klebsiella pneumoniae was the main epidemic clone of carbapenem-resistant Klebsiella pneumoniae (CRKP). The 90 kb size plasmid was the main plasmid encoding blaKPC-2 gene. Carbapenemase genes can be transferred horizontally through plasmids. The hospital should strengthen prevention of nosocomial infections to control the clonal prevalence of CRE.