Objective To explore the clinical characteristics and diagnosis of Epstein-Barr(EB) virus encephalitis(EBE) in children. Methods The verification of EBE was based on detection of EBV DNA in cerebrospinal fluid (CSF) by fluorogenic quantitative PCR(FQ-PCR) and Nest-PCR. The clinical and CSF changes of 27 EBE cases and 26 controls were analyzed and compared. Results EB-DNA in CSF by FQ-PCR of 13 cases of EBE was (2.82?2.03)?10 3copies.There was no significant difference in clinical manifestations between EBE and HSE groups, except that WBC in CSF of EBE was lower than those of HSE.Conclusions EBE is not infrequent, about 10 % of encephalitis in children. EBE is always present independently, which is not a complication of infectious mononucleosis(IM).Detection of EBV-DNA in CSF is a sensitive and specific test for diagnosing of EBE. Early treatment may be beneficial to the prognosis of EBE.

Objective To analyze the genetic characteristics of breakthrough varicella-zoster virus infection during the varicella outbreaks in Minhang District of Shanghai in 2013-2014. Methods Samples of the varicella-zoster liquid were collected from patients with chickenpox in Minhang District in 2013-2014 and used for the extraction of genomic DNA. The open reading frame ( ORF) of 22 and 62 regions were se-quenced and further analyzed by using bioinformatics methods. Results A total of 24 samples were success-fully collected and sequenced, and all of them were wild strains. Among the 24 varicella-zoster virus strains, 23 strains were highly homologous to the parental strain ( P-Oka) and the vaccine strain ( V-Oka) , indica-ting that they belonged to J genotype. Only one strain, whose genotype was between M and E, was highly ho-mologous to the mosaic( M) CA123 strain ( M1 genotype) , indicating that it might belong to M1 genotype. Conclusion The epidemic strains of varicella-zoster virus in Minhang District were mainly J genotype. Lo-cal epidemic of M and other genotypes of varicella-zoster virus also existed. There were some gene variations in different strains of J genotype. These varicella-zoster virus strains of non-vaccine genotypes might be one of the reasons causing the breakthrough cases of chicken pox.

Objective To evaluate the epidemiological efficacy of 23-valent pneumococcal poly-saccharide vaccine on the clinical symptoms and the incidence of respiratory tract infections in the elderly over 60 years old in Minhang District, Shanghai. Methods A prospective cohort study was conduct, in which the sample size was 1 200 for both inoculated and uninoculated groups. This study followed the two groups of subjects every quarter for a year. Clinical symptoms and the incidence of respiratory tract infectious diseases as well as the acute onset of chronic respiratory diseases were compared between the subjects of the two groups. Results Compared with the uninoculated group, less people in the inoculated group developed the clinical symptoms of respiratory tract infection (RR=0. 894, 95% CI: 0. 804-0. 994). Immunization with the 23-valent pneumococcal polysaccharide vaccine protected 57. 7% of the senior citizens (≥75 years old) from respiratory tract infections(95% CI:0. 207-0. 775). Conclusion The 23-valent pneumococcal polysaccharide vaccine can effectively reduce the incidence of respiratory tract infections in the elderly aged 60 years old and over and protects the elderly aged 75 years old and over from respiratory tract infections. Further studies on the immunological efficacy of 23-valent pneumococcal polysaccharide vaccine should be carried out by measuring the antibody titers before and after vaccination.

Objective To compare the sub-cellular proteome of isoniazid ( INH)-resistant Mycobacterium tuberculosis (MTB) with that of sensitive strains for identifying of unique proteins of these strains and discussing their preliminary application in clinical diagnosis. MethodsProteins of cell wall and membrane of 5 INH-resistant strains and 5 INH-sensitive strains were extracted by density gradient centrifugation.The extracts were subsequently analyzed using weak cation exchange (WCX) liquid chromatography ( LC )followed reverse phase (RP) liquid chromatography to compare the sub-cellular protein patterns. A total of 1280 fractions were collected and identified by matrix assisted laser desorption ionization-time of flight-tandem mass spectrometry (MALDI-TOF MS/MS). The Database for Annotation, Visualization and Integrated Discovery (DAVID) was used for cell component and biological process analysis. Normalized Spectral Abundance Factors (NASF) was used for semi-quantity of protein expression. 5 proteins significantly up-regulated in INH-resistant strains and 2 proteins significantly up-regulated in INH-sensitive strains were selected for ELISA analysis with autologous sera respectively. Results A total of 347 proteins were identified. Cell component analysis showed that 58％ proteins were cells well or membrane proteins. Biological process analysis showed that 31％ proteins involved in carboxylic/monocarboxylic acid biosynthetic and metabolic process, 26％ and 15％ proteins involved in organic acid or fatty acid biosynthetic and metabolic process,while 28％ proteins involved in lipid biosynthetic , metabolic, transport and localization process. O-succinylbenzoate synthase, monooxygenase, hypothetical protein Rv2255c, nicotinate-nucleotide--dimethylbenzimidazole phosphoribosyltransferase and membrane phosphatidate cytidylyltransferase cdsA were up-regulated in INH-resistant strains and fractions contained these proteins could elicit specific antibody response with autologous sera. The A450 was higher than that with INH-sensitive sera. The differences between the INH-resistant sera and the INH-sensitive sera were significant ( t = 0.028, 0.044, 0.066, 0.064, 0.083, all P＜0.01 ). Chain A of Rv2002 Gene Product and Chain A of Crystal Structure Of Rv2632c were up-regulated in INH-sensitive strains and fractions contained these proteins could elicit specific antibody response with autologous sera. The A450 was higher than that with INH-resistant sera. The differences between the INH-sensitive sera and the INH-resistant sera were significant (t=0.053, 0.073, both P＜0.05). ConclusionThe combination of density gradient centrifugation and 2D-LC MS/MS technology is useful in enrichment and identification of differential expressed proteins between INH-resistant and INH-sensitive strains at sub-cellular level. It is useful in finding antigens associated with INH-resistant MTB infection, which may prove useful for further study in the mechanism of INH resistant, as well as interaction between MTB and host.

Objective To evaluate the efficacy and safety of colistimethate sodium (CMS) for the treatment of critical patients infected by pan-drug resistantAcinetobacter baumannii (PDR-AB) or pan-drug resistant Pseudomonas aeruginosa (PDR-PA).Methods 321 isolates of PDR-AB and 204 isolates of PDR-PA from critical patients admitted to 35 intensive care units (ICUs) of grade two or above were collected from the Anhui Antimicrobial Resistance Investigation Net (AHARIN) program from September 2012 to September 2015, while the minimal inhibitory concentrations (MIC) of colistin were determined by the E-test. A series of Monte Carlo simulations was performed for CMS regimens (1 MU q8h, 2 MU q8h, and 3 MU q8h, and MU meant a million of unit), and the probability of achieving a 24-hour area under the drug concentration time curve (AUC24)/MIC ratio > 60 and risk of nephrotoxicity for each dosing regimen was calculated. Each simulation was run over three CLCr ranges: < 60, ≥ 60-90, ≥ 90-120 mL/min. The probability of target attainment (PTA)for the AUC24/MIC ratio was calculated using the partial MIC value, while the cumulative fraction of response (CFR) was determined by integrating each PTA with the MIC distributions, the value greater than or equal to 90% or more than 80% was set as the optimal dosing regimen or suboptimal dosing regimen respectively. The probability of average 24-hour serum concentrations up to 4 mg/L for three dosage regimens was used to predict the risks of nephrotoxicity.Results All 321 isolates of PDR-AB and 204 isolates of PDR-PA were susceptible to colistin, the MIC50/90 against PDR-AB were 0.5mg/L and 1.0 mg/L, and those against PDR-PA were 0.5 mg/L and 1.5 mg/L, respectively. When recommended dose (1 MU q8h) was used for patients with CLCr of < 60 mL/min, high CFR value (89.78% for PDR-AB, 81.06% for PDR-PA) were obtained, but with a high risks of nephrotoxicity (> 32.51%). Moreover, low value of PTA (< 66.56%) was yielded for isolates with MIC of ≥ 1 mg/L. Recommended dose also yielded a low CFR value (56.97%-69.31% for PDR-AB, 44.76%-56.94% for PDR-PA) in patients with CLCr of ≥ 60-120 mL/min. When dose was increased to 2 MU q8h, CFR (77.45%-92.87%) and the risks of nephrotoxicity (< 0.15%) was optimal for patients with CLCr ≥ 60-120 mL/min, but low value of PTA (< 75.36%) was also yielded for isolates with MIC of ≥ 1 mg/L. The most aggressive dose of 3 MU q8h provided high CFR (> 89.24%) even in patients with CLCr ≥ 90-120 mL/min, and PTA was < 76.20% only for isolates with MIC of ≥ 1.5 mg/L, but this dosing scheme was associated with unacceptable risks of nephrotoxicity (> 33.68%).Conclusion Measurement of MIC, individualized CMS therapy and therapeutic drug-level monitoring should be considered to achieve the optimal drug exposure and ensure the safety of CMS.

Objective To analyze a population of cells on the right lower lateral of monocyte population in forward scatter/side scatter(FSC/SSC)(X-axis/Y-axis)scatterplot of peripheral blood leucocyte by flow cytometry(FCM)and its influencing factors.Methods The type of cells were identified based on cluster of differentiation antigen(CD)by FCM.The impact of temperature,hemolysin concentration,and incubation time was evaluated.Blood lipid tests were performed to observe the relation between them by statistical methods.Results (1) Phenotypo of this population of cells on the right lower lateral of monocytes in FSc/SSC scatterplot is CD+45 CD+13 CD+14 CD3- CD-19 ,which was the same as monocyte cells:(2)The monocytes in FSC/SSC scatterplot shifted to left side after using haemolysin;(3)The monocytes showed less resistance to antihemolysin in 37℃ than that in 220C:There were more monocytes shifting to left side with the increase of haemolysis time:(4)The swarming ratio of monocytes in patients (31.5%,40/126) Was higher than it in normal controls (5.1%,5/98)(x2=22.74,P<0.01);(5)The levels of total serum cholesterol(TC),triglyeride(TG),low density lipoprotein cholesterol(LDL-C), apoprotein B100(Apo B100) in patients with swarming monocytes were lower than that in the patients without swarming monocytes,(P<0.05).There was no statistical significance between the two groups with respect to levels of total bilirubin(TBIL),albumin(Alb),hish density lipoprotein cholesterol(HDL-C),apoprotein A I(Apo A I),lipoprotein(Lpa).Conclusions Peripheral blood monocytes can be divided in two groups in FSC/SSC scatterplot when analyzed with FCM.The presence of this population of cell Was related to resistance to hemolysin.It can be influenced by haemolysis time and incubation temperature.Therefore,the effect of swarming monocytes and abnormal cell membrane should be taken into consideration when the markers and function of monocytes are detected by FCM.

Objective To identify the success rate of peripheral indwelling needle puncture in pediatric patients, and analyze its influencing factors. Methods A survey was conducted in a sample of 902 pediatric patients.Personal information table of pediatric patients who receive infusionwas adopted to collect data. Results The success rate of first puncture was 85.37%(770/902), the success rate of two punctures was 95.34%(860/902). The success rate was affected by several factors, such as department, ages of the patients, condition of the veins, years of working as a nurse(OR=1.128, 2.308, 2.351, P <0.05). Conclusions Success rate of peripheral indwelling needle puncture in children still can be improved. When formulate management program, the influencing factors should be considered for management standard.

Objective To measure the titers of varicella live attenuated vaccines ( VarV) during the process of transportation and storage in different seasons and communities of Minhang Distract, Shang-hai, to evaluate the operation of cold chain system and the thermal stability of vaccines and to provide refer-ences for the management, introduction, research and development of vaccine in future.Methods Four communities with high outbreak rates of varicella during 2012 to 2013 and four communities with low out-break rates were randomly selected from 13 communities in Minhang District, Shanghai.The titers of VarVs were detected by using the quantitative plaque assay before and after 30 days of storage in November 2013 and February, May, August 2014.Results The overall rate of qualified VarVs was 90.63% with a geo-metric mean titer (GMT) of 3.67 (LgPFU/0.5 ml).96.88%of the VarVs produced by company C met the quality standard with a GMT of 3.89 ( LgPFU/0.5 ml) followed by those produced by company B with a rate of 91.67%and a GMT of 3.75 ( LgPFU/0.5 ml) .The rate of qualified VarVs produced by company A was the lowest, which was 80.00%, with GMT of 3.29 (LgPFU/0.5 ml).There were significant differences in the rates of qualified VarV among the three companies (χ2=8.288, P=0.016).The rate of qualified vac-cines in communities close to the Shanghai Minhang Center for Disease Control and Prevention ( Minhang CDC) was 91.67%, while 100%of the vaccine samples collected form the communities at a middling dis-tance from or far from the Minhang CDC met the quality standard.No significant difference in the rate of qualified vaccine was found among the three types of communities (χ2=3.441, P=0.179).The quarterly rates of qualified vaccines produced by B and C companies were 100%except for the third quarter.The rates of qualified vaccines produced by A, B and C companies in the third quarter were respectively 70.00%, 66.67%and 87.50%.No statistical differences in the quarterly rates of qualified vaccines were found among the three companies (χ2=1.25, P=0.7412;χ2=6.545, P=0.088; χ2=6.194, P=0.103).No statistical differences in the rates of qualified vaccines before and after 30 days of storage were found among the three companies (χ2=0.625, P=0.347;χ2=0.000, P=1.000;χ2=2.065, P=0.492).Conclusion A well-managed and-operated cold chain system was implemented in Minhang District in the storage and transport of vaccines as well as other related links.The thermal stability of vaccines produced by company C was better than those of the other two companies.A surveillance for the titers of vaccines produced by com-pany A should be strengthened.

Salvia miltiorrhiza is a highly valued traditional chinese medicine for the treatment of atherosclerosis-related disorders in china, such as cardiovascular and cerebrovascular diseases in China. The wilt disease is serious in the culture of S. miltiorrhiza. Wilt disease cause biomass of plant shoots and roots is lessened, active components are decreased. To solve these problems, we research the pathogen causing wilt disease of S. miltiorrhiza. The suspected pathogen is identified by morphology and etiological test. The identification was further confirmed by alignment the sequences of internal transcribed spacer (ITS) amplified by PCR. Our result show the wilt disease of S. miltiorrhiza mostly occurred in July and August, which is hot and wetter. The wilt disease rate of S. miltiorrhiza continuous cropping for one year in S. miltiorrhiz stubble is 10%, but the wilt disease rate of S. miltiorrhiza continuous cropping for three years in S. miltiorrhiz stubble is 60%-70%. The root rot of S. miltiorrhiz caused by the wilt disease, so the wilt disease was mistaken for the rot root in production. Morphological characteristics show the pathogen is Fusarium oxysporum. The sequence of ITS wes determined and found by BLAST shared 99% identity to that of F. oxysporum f. sp. cucumerinum. So it comes to the conclusion that the causing agent of wilt disease on S. miltiorrhiza belongs to F. oxysporum.
DNA, Intergenic ; genetics ; Fusarium ; genetics ; isolation & purification ; physiology ; Plant Diseases ; microbiology ; Polymerase Chain Reaction ; Salvia miltiorrhiza ; microbiology ; Seasons

OBJECTIVETo measure the threshold of feeling temperature, comfortable temperature and tolerant temperature to ginger-partition moxibustion and suspended moxibustion at different acupoints of healthy persons, and to instruct the clinical practice of moxibustion better.

METHODSForty-three healthy students were divided into a ginger-partition moxibustion group (22 cases) and a suspended moxibustion group (21 cases) randomly. Shenshu (BL 23), Zhongwan (CV 12), Zusanli (ST 36) and Waiguan (TE 5) were used in both groups. The feeling temperature, comfortable temperature and tolerant temperature in each group were measured by thermometer. SPSS 18.0 was used to analyze the data.

RESULTSThe feeling temperature, comfortable temperature and tolerant temperature rose in proper order and the difference was significant (all P < 0.05). The tolerant temperature of suspended moxibustion group ((40.69 +/- 1.85) degrees C) was lower than that of ginger-partition moxibustion group ((42.44 +/- 3.90) degrees C) (P < 0.05). The feeling temperature of ginger-partition moxibustion at Shengshu (BL 23) was 4-5 degrees C lower than that at other acupoints.

CONCLUSIONThe heat sensations of different acupoints to ginger partition moxibustion and suspended moxibustion are different at different stages. The threshold of heat sensation of ginger-partition moxibustion is higher than that of suspended moxibustion. There is a best temperature zone for moxibustion which is a little higher than normal temperature between comfortable temperature and tolerant temperature. To improve effect, doctors should try to prolong the effective time of this zone in clinic. As the limiting temperature of patients, tolerant temperature can provide reference for the use of moxibustion safely in clinic.