Effects of severe fever with thrombocytopenia syndrome virus infection on the expression of TLR2 and inflammatory cytokines in mouse peritoneal macrophages / 医学研究生学报

Objective The secretion level of inflammatory factors is closely related to the severity of severe fever with thrombocytopenia syndrome (SFTS). This paper mainly discussed the effect of SFTSV infection on the expression of Toll-like receptor 2（TLR2） and inflammatory factors in macrophages in mice. Methods The expression of TLR2 andTNF-α, IFN-β, IL-6, IL-10 and other inflammatory factors were observed at 0, 6, 12, 24, 36, 48, 72 hours after SFTSV infected mice macrophages by qPCR, Western blot and ELISA. Results QPCR results showed that the mRNA levels of TNF-α at 0, 6, 12, 24, 36, 48, 72 h were 0.85±0.14, 15.23±2.45, 28.67±1.59, 94.52±7.05, 55.86±1.82, 23.55±6.15, 9.76±2.03, and the mRNA levels of IFN-β were 0.93±0.21, 1.55±0.33, 14.51±1.98, 55.16±3.64, 26.57±1.49, 9.22±0.51, 5.18±1.06. QPCR results showed that the mRNA levels of TNF-α and IFN-β produced by abdominal macrophages in the infected mice showed a trend of first increasing (the highest at the 24 hour) and then decreasing, and the difference of TNF-α and IFN-β mRNA levels at other time points was statistically significant compared with that at the 0 hour (P<0.05). However, IL-6 and IL-10 mRNA levels continued to increase, and the difference at other time points was statistically significant (P<0.05). ELISA results showed that the expression of four inflammatory factors showed a trend of gradual increase: TNF-α 0 and 72 h were (38.31±4.25, 140.41±23.45) pg/mL; IFN-β 0th and 72th were (17.56±0.66, 1084.93±111.42) pg/mL; IL-6 protein 0 and 72 h were (113.30±0.07, 2302.32±134.09) pg/mL; IL-10 protein 0 and 72 h were (515.00±21.21, 2590.40±226.19) pg/mL, respectively, with significant increases at the 24, 36, 48 and 72 hours compared with that at the 0 hour (P<0.05). The expression of TLR2 mRNA generated by mouse peritoneal macrophages showed a trend of first increasing and then decreasing, and increased to the highest level at 24 h, and the difference between each time point and 0 h was statistically significant (P<0.01). The expression of TLR2 gradually increased after infection with time extension. Conclusion SFTSV infection can up-regulate the expression of TLR2 in macrophages, thereby leading to the increased secretion of the cytokines.

Prokaryotic expression and preparation of the polyclonal antibodies of E protein and domain Ⅲ in Zika virus / 中国人兽共患病学报

To clone,express and purify the E Protein and EDⅢ of Zika virus in E.coli and prepare two kinds of polyclonal antibodies,the virus was amplified by Vero E6 cell culture.Total RNA was extracted by RT-PCR and reverse transcribed into cDNA.The prokaryotic expression vectors pET32a/E and pET28a/EDⅢ were constructed by cDNA sequence of E and EDⅢ gene.Then,recombinant plasmids were transformed into E.coli BL21 and induced by IPTG,and purified by Ni+ column affinity chromatography.BALB/C mice were immunized with purified recombinant proteins.Antiserum was collected and titer was determined by indirect ELISA.Western blot was used to detect the specificity.Results showed that the recombinant proteins were successfully expressed and purified.The titer of the polyclonal antibodies both reached 1:409 600.Western Blot analysis showed that the polyclonal antibodies could specifically recognize the recombinant proteins.Thus,the specific polyclonal antibody were successfully prepared,laying a foundation for further study on the pathogenesis,detection methods and immune strategies of Zika virus.

An investigation on adult dietary protein intake in Zhejiang Province / 预防医学

Objective TolearntheproteinintakestatusofadultsinZhejiangProvinceandtoinvestigatetherelationship between the protein intake and influencing factors,and in order to provide a scientific basis for improving adult protein intake.Methods Datawereselectedfromthe2010—2012ChineseNationalNutritionandHealthSurveyinZhejiang province.Data were gained through medical examination and the method of 3 day 24-hour dietary recall and food weighted record.Descriptiveanalysiswasconducted.Results Therewere1160men(48.09%)and1252women(51.91%)in 2 412 cases in this analysis.The average protein intake per day was 71.87 g for per reference man,and the intake quartile was 66.06(51.17-85.93)g/d and 38.14% of adults were not achieved the Recommended Nutrient Intake (RNI).The multi factor logistic regression analysis showed that age,region,income and occupation were the main factors affecting protein intake.Age was a risk factor,while income was a protective factor.Big cities were more easily to have insufficient protein intake than median and small cities.Students,agriculture,housework and other groups of adults were more easily to have insufficient protein intake.And 30.70% of dietary protein was from cereal,and 39.70% was from animal food. Sources of dietary protein were statistical significant among different areas and age.Protein intake from cereal among young people (29.60%),old people (29.95%),and people living in big cities (19.81%)was low.Protein intake from cereal among people living in medium and small cities (10.40%)was high.Young people (43.12%)and people living in big cities(52.87%)hadhigheranimalsourceprotein.Conclusion TheproteinintakeofadultsinZhejiangProvincewasnot achieved the RNI.It is important to conduct health educations according to the protein intake problems of different groups to improve their protein intake status.

Viral and Bacterial Etiology of Acute Diarrhea among Children under 5 Years of Age in Wuhan, China / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Acute diarrhea remains the serious problem in developing countries, especially among children under 5 years of age. Currently, only two or three common diarrhea pathogens were screened at most hospitals in China. The aim of this study was to provide a wide variety of diarrhea pathogens and their antimicrobial resistance patterns in children under 5 years of age.</p><p><b>METHODS</b>Totally 381 stool samples collected from Tongji Hospital between July 1, 2014 and June 30, 2015 were tested by culture and/or polymerase chain reaction for eight kinds of bacteria and five kinds of viruses. An antimicrobial sensitivity test was performed using dilution method recommended by the Clinical and Laboratory Standards Institute.</p><p><b>RESULTS</b>Viral infections were mainly identified in infants (0-11 months), whereas bacterial infections were more prevalent in the age of 24-59 months. About 69.8% of samples were positive for at least one pathogen, 51.7% of samples were virus positive, followed by bacteria positive cases (19.4%), and 12.6% of cases displayed co-infections with two viruses or a virus and a bacterium. Rotavirus was the most prevalent pathogen, followed closely by norovirus, while Salmonella was the most commonly isolated bacteria, followed by diarrheagenic Escherichia coli (DEC) and Campylobacter. More than 40% of Salmonella spp. and DEC isolates were resistant to first-line antibiotics (ampicillin, trimethoprim-sulfamethoxazole, and tetracycline). Around 10% of Salmonella spp. isolates were resistant to ceftriaxone and ciprofloxacin simultaneously. Campylobacter spp. displayed high resistance to ciprofloxacin but kept low resistance to azithromycin and doxycycline.</p><p><b>CONCLUSIONS</b>The etiology of acute diarrhea varies in children of different age groups. The high frequency of infection with viruses suggests the urgent demand for new viral vaccine development. Proper use of antibiotics in the treatment of acute diarrhea is crucial due to the high level of antibiotic resistance.</p>

Dietary Patterns Associated Metabolic Syndrome in Chinese Adults / 生物医学与环境科学（英文）

Dietary pattern has been revealed to be associated with metabolic syndrome. However, the association was not well documented in Chinese due to the complexity of Chinese foods. We mainly assessed the dietary patterns and examined their effects on metabolic syndrome among Chinese adults. Four dietary patterns including 'Refined Grains & Vegetables' Pattern, 'Dairy & Eggs' Pattern, 'Organ Meat & Poultry' Pattern, and 'Coarse Grains & Beans' Pattern were extracted. 'Dairy & Eggs' Pattern was associated with a decreased odds of metabolic syndrome in women, and 'Coarse Grains & Beans' Pattern was associated with a decreased odds of hypertension in men. These results provided a scientific basis for future research and dietary guideline perfection.

Epidemiological study of Streptococcus pneumoniae in the nasopharynx of healthy children under 5 years of age in Wuhan / 中华儿科杂志

<p><b>OBJECTIVE</b>To investigate the population biology of Streptococcus (S.) pneumoniae carried by healthy children under 5 years of age in Wuhan.</p><p><b>METHODS</b>S. pneumoniae was isolated from nasopharyngeal swabs of healthy children under the age of 5 years (under 5) in Wuhan. The susceptibility to 12 antimicrobial agents was tested by agar dilution method. The erythromycin resistant genes were detected by using the technique of polymerase chain reaction. The quellung reaction was used for serotyping.</p><p><b>RESULTS</b>The carrier rate of S. pneumoniae was 22.31% (135/605) in under 5 children in Wuhan. Among the 133 alive strains, the proportion of penicillin non-susceptible S. pneumoniae (PNSSP) was 45.9% (61/133). The susceptibility of S. pneumoniae to the first (cefalexin), the second (cefaclor) and the third (cefixime, cefpodoxime and cetriaxone) generations of cephalosporins was 6.0%, 45.1%, 54.9%, 56.4%, and 88.7%, respectively. None of the strains were found resistant to fluoroquinolones except for one strain which was resistant to ciprofloxacin at low level. The susceptibility to macrolides was low, 14.3% - 15.8%. Among totally 114 strains resistant to erythromycin, the genes ermB alone, both ermB and mefA, and mefA alone were found in 76 strains (66.7%), 46 strains (40.4%) and 2 strains (1.8%), respectively. Seventeen serotypes were involved and the prevalent serotypes were 19, 23, 6, 15 and 14. Seven strains remained nontypable for serotype. PNSSP strains were found to be distributed in serotypes 19, 23, 6 and nontypable group.</p><p><b>CONCLUSIONS</b>The antibiotic resistance of S. pneumoniae was serious in Wuhan area. The ribosomal modification (ermB gene mediated) was the main mechanism of S. pneumoniae resistant to erythromycin. The major prevalent serotypes were 19, 23 and 6.</p>

Analysis of the distribution characteristics and drug sensitivity of infection pathogens in hematology department ward / 中国实验血液学杂志

The objective of this study was to investigate clinical distribution characteristics and drug sensitivity of infection pathogens in hematology department wards of our hospital during recent two years. The isolation and cultivation of pathogens from samples were performed by routine methods. Drug sensitivity tests of bacteria were performed by Kirby-Bauer method. Drug sensitivity tests of fungi were performed by ATBFUNGUS Drug sensitivity strips. The results showed that 102 strains of pathogens were isolated from all detected samples. The composition ratio of Gram-positive bacteria, Gram-negative bacteria and fungi was 42.2%, 34.3%, 3.5%, respectively. 58.8% of pathogens were isolated from samples of malignant hematopathy patients. 27.5% were isolated from samples of the patients with fever of unknown origin (FUO). 51.0% of pathogens were isolated from samples of the patients who suffered from agranulocytosis or leucocytopenia. Isolated fungi were mostly sensitive to anti-fungal drugs. G+ bacteria were most sensitive to vancomycin. G- bacteria were most sensitive to imipenem. Most bacteria were resistant to multiple antibiotics. It is concluded that the infection in hematology department wards was related with many conditions, such as weakened resistance of patients, leucocytopenia or agranulocytosis, tumor loading, etc. The prompt microbiological examination and drug sensitivity tests are important to rationally select antibiotics, reduce infection incidence and mortality rate, and decrease the occurrence of drug resistant strains.

Studies on drug resistance genes among 49 strains of extended-spectrum beta-lactamase-producing bacteria / 中华检验医学杂志

Objective To investigate the drug resistance genes of extended-spectrum beta- lactamase-producing bacteria in 49 strains.Methods Extended-spectrum ?-lactamase -producing strains were detected by the disc diffusion test.The techniques of polymerase chain reaction,sequence analysis, pulsed-field gel electrophoresis were used to analyze the genotype and homology of extended-spectrum ?- lactamase-producing strains.Results The incidence of ESBL-producing strains from E.coli,K pneumoniae,K oxytoca,was 20% in 2000,and 40% in 2003.Among the 49 ESBLs producers the most common genotype was CTX-M-14( n=33).The others were CTX-M-3,CTX-M-9,CTX-M-12,CTX-M-15, CTX-M-24 and SHVSa.Both two CTX-M subtypes,CTX-M-3 and CTX-M-14,were detected in one strain. However,4 ESBL-producing strains confirmed by phenotype remained untyped.The results showed that the ESBLs producers were not closely related,except for two strains of E.coli and two strains of K.pneumoniae which were homgenic respectively.Concolusions The incidence of ESBL-producing strains increases with years.The most common genotype of ESBLs is CTX-M.There is no evidence for epidemiologic spread of ESBL-producers by pulsed-field gel electrophoresis.