Expression of long noncoding RNA in papillary thyroid carcinoma with Hashimoto's thyroiditis / 中华内分泌外科杂志

Objective To analyze long noncoding RNAs (lncRNAs) expression profiles in papillary thyroid carcinoma (PTC) with Hashimoto's thyroiditis (PTC-HT,group A) and PTC only (PTC,group B).Methods 55 cases of thyroid species were collected.High-throughput microarray lncRNh was used to detect the expression difference of lncRNAs between group A and group B.Real-time quantitative PCR (QRT-PCR) was used to verify.Results 1031 lncRNAs and 1338 mRNAs were detected abnormally expressed in tissue samples of group A compared to B.GO and Pathway analysis of mRNAs suggested some biological processes changed obviously,such as immune system and immune reaction.QRT-PCR showed that the expression of uc002stn.1,ENST00000452578 and uc002sti.1 in group A and group B was significantly different.Conclusion IncRNAs expression was significantly different in PTC with or without HT,which may play important roles in the pathogenesis of PTC with HT.

Advances of long non-coding RNA in thyroid tumor / 中华内分泌外科杂志

Long non-coding RNA(lncRNA) is non-protein coding transcripts longer than 200 nucleotides,which plays an important role in the development of the metabolic process.Thyroid cancer is the most common cancer of the endocrine system,and as reported,lncRNA is related to the occurrence and development of thyroid tumors.Therefore,this paper reports the latest domestic and foreign research progress about lncRNA in thyroid tumor,in order to provide new ideas for molecular diagnosis and treatment of thyroid cancer.

Determination on the Content of Copper and Zinc in Tea by Flame Atomic Absorption Spectrophotometry / 沈阳医学院学报

Objective:To determine the content of copper and zinc in tea by flame atomic absorption spectrophotometry and to investigate the influence on the content after leached for different time. Methods:The tea samples were leached for different time by boiling water and then the content of copper and zinc was determinated by flame atomic absorption spectrophotometry. Results:When the leaching time was less than 10 min, the content of copper and zinc increased significantly with the prolongation of the leaching time, while the increase was not obvious when the leaching time was more than 10 min. Copper and zinc showed good linearity in the range of 0.02-0.15 mg/L and 0.05-0.30 mg/L,separately. The recovery was in the range of 98.5%-101.1%for copper and 98.7%-101.2%for zinc with the RSD of 0.8%in copper and zinc (n=9) . Conclusion:The method is suitable to determine the content of copper and zinc in tea. Leaching time can influence the content of copper and zinc in tea significantly.

Comparison of six methods for fungal genomic DNA extraction / 中华临床感染病杂志

Objective To identify a rapid and efficient fungal genomic DNA extraction method for PCR amplification.Methods Genomic DNA was extracted from Penicillum marneffei,Rhizopus microsporus,Cryptococcus neoformans and Candida albicans by heating pyrolysis,microwave,repeated freezing and thawing,lysozyme digestion,overnight snail enzymatic and Qiagen kit methods.DNA electrophoretogram was observed by gel imaging system.The concentration and purity of extracted DNA were determined with an ultramicro nucleic acid protein tester and the yields were calculated.PCR amplification and sequencing were also performed.ANOVA and SNK-q test were used for data analysis.Results There were statistical differences in concentrations and yields of the fungal DNA extracted from Penicillum marneffei (hyphal phase and yeast phase),Rhizopus microsporus,Coptococcus neoformans and Candida albicans by six methods (F=750.83,220.95,669.35,132.01,510.20 and 1658.35,287.10,963.64,1147.77,4521.22,all P ＜0.01).Of six methods,microwave method gained the highest DNA concentration and yield,followed by heating pyrolysis method,while Qiagen kit method obtained the lowest concentration and yield.All DNA extracted by 6 kinds of methods were positive in PCR amplification.Conclusion All of the six methods can be used for fungal DNA extraction which is sufficient for PCR amplification,but microwave and heating pyrolysis methods are more easy and simple to perform.

Epidemiological characteristics of mycoplasma pneumoniae, Epstein-Barr virus and cytomegalovirus in children with acute upper respiratory tract infections in Hangzhou / 中华临床感染病杂志

Objective To investigate the epidemiological characteristics of mycoplasma pneumoniae (MP),Epstein-Barr virus (EB virus) and cytomegalovirus (CMV) infection in children with acute upper respiratory tract infections in Hangzhou.Methods Throat swabs and sputum samples were collected from 5942 children with acute upper respiratory tract infections in Hangzhou First People's Hospital during January 2011 and December 2012.MP,EB virus and CMV were detected using quantitative PCR.The distribution and seasonal changes of the above pathogens in children of different ages were analyzed using Chi-square tests.Results MP,EB virus and CMV were positive in 29.91％ (1777/5942),22.92％ (1362/5942) and 8.55％ (508/5942) children,respectively.Mixed infections were found in 556 (9.36％) children.The positive rates of MP varied among different age groups (x2 =113,P =0.000),and the highest one was detected in children ＞ 6-year old (448/1012,44.36％).EB virus infection was rare in age group 0-1 year,and the positive rate was of statistical difference from those in other age groups (x2 =167,181 and 187,P =0.000).The highest positive rate of CMV (23.78％) was found in children aged 0-1 year old.The positive rates of MP varied in different months of the year (x2 =208 and 211,P =0.000),and the highest positive rate was found in July and August.Conclusion The predominant pathogen of acute upper respiratory tract infection in children is MP in Hangzhou,and MP plus EB virus infection is common,particularly in older children;while CMV infection more likely occures in 0-1 year old babies,and usually in summer.

Establishment and application of DPO-based quadruple real-time PCR for detection of mycobacteria / 中华检验医学杂志

Objective To establish a rapid,accurate and specific method to detect the common mycobacteria based on multiplex real-time PCR.Methods The dual priming oligonucleotide ( DPO)primers and TaqMan probes labeled with FAM,ROX,HEX or JOE fluoresceins at 5' end and eclipse at 3' end respectively were designed to detect the 16S rRNA of mycobacteria.Both specificity and sensitivity were estimated on multiplex real-time PCR detecting genome DNA from 4 mycobacterial model species.Sixty eight early morning sputum specimens collected from hospitalized patients in the Red Cross Hospital of Hangzhou were detected by multiplex real-time PCR,bacterial culture and smear microscopy simultaneously.The positive rates were analyzed by chi-square.Results Mycobacteria including Mycobacterium tuberculosis and three common non-tuberculosis mycobacteria spp.were identified by multiplex real-time PCR accurately and specifically,with the limited load at 101 cfu/ml.In 68 sputum specimens,31 were positive (positive rate 45.6％ ) by this method,which was significant higher than that by smear microscopy ( positive rate 14.7％,x2 =15.4,P ＜0.05 ).The positive cases were identified as 28 Mycobacterium tuberculosis,1 Mycobacterium avium and 2 Mycobacterium intracellulare in agreement with the culture results.One case,which is detected by culture,but not by PCR,was identified as Mycobacterium chelonae by sequencing.Conclusion The multiplex real-time PCR characterizing as sensitive,specific and time-saving for Mycobacterium tuberculosis and common non-tuberculosis mycobacteria could be chosen as the rapid laboratory test of mycobacterial infection.

Simultaneous detection of four types of human herpes virus by AllGlo quadruple fluorescence quantitatve PCR / 中华微生物学和免疫学杂志

ObjectiveTo optimize the condition of multiple fluorescence quantitative PCR and establish a new assay of four human herpes virus (HHV) detected by AllGlo quadruple fluorescence quantitative PCR.MethodsFour HHV including HSV-1,HSV-2,EBV and CMV were identified by sequence analysing the qualitative PCR production.Furthermore,they were quantitatively detected by AllGlo and TaqMan multiple fluorescence quantitative PCR respectively.ResultsBoth the positive rate and specificity of AllGlo and TaqMan in detecting single HHV achieved 100％.And AllGlo single fluorescence quantitative PCR prevailed over TaqMan's by Ct of 1-3.Four HHV can be simultaneously detected by AllGlo quadruple fluorescence quantitative PCR,comparing to the only two by TaqMan.ConclusionAllGlo fluorescence quantitative PCR assay allows a higher throughput,sensitivity and specificity than TaqMan in detection and thus provides a board prospect.

HBV subgenotype C2 infection, A1762T/G1764A mutations may contribute to hepatocellular carcinoma with cirrhosis in southeast China

To glean insights into the relationship among hepatitis B virus [HBV] genotype/subgenotypes, A1762T/G1764A mutations and advanced liver disease such as liver cirrhosis [LC] and hepatocellular carcinoma [HCC] in Southeast China. Methods: A case-control study was performed, consisting of chronic hepatitis B [CHB] patients [n=160], LC patients [n=150], and HCC patients [n=156]. Fluorescence quantitative polymerase chain reaction [FQ-PCR] was used to detect A1762T/G1764A mutations. HBV genotypes/subgenotypes were determined by multiplex PCR. All patients' clinical data was systematically collected from the hospital records. Results: Our study revealed HBV genotypes C [63.95%] and B [33.69%] were predominant in chronically infected patients, subgenotype B2, C2 and C1 were the major subgenotypes. Both subgenotype C2 infection and A1762T/G1764A mutations were associated with LC and HCC with cirrhosis, subgenotype C2 [OR=2.033, 95%CI=1.246-3.323, P=0.003 for LC vs CHB; OR=3.247, 95%CI=1.742-6.096, P=0.001 for HCC with cirrhosis vs CHB; respectively], and A1762T/G1764A mutations [OR=1.914, 95%CI=1.188-3.085, P=0.005 for LC vs CHB; OR=2.996, 95%CI=1.683-5.353, P=0.002 for HCC with cirrhosis vs CHB; respectively], but no differences in the frequencies of both variants between LC and HCC with cirrhosis groups were found. Conclusions: HBV subgenotype C2 infection and A1762T/G1764A mutations are both risk factors of LC and HCC with cirrhosis development in the patients with CHB in Southeast China, but all no helpful for predicting HCC development in LC patients

Fluorescence PCR for detecting ureA gene of Helicobacter pylori in feces / 中华临床感染病杂志

Objective To evaluate the application of fluorescence PCR in detecting ureA gene of Helicobacter pylori(HP)in feces.Methods Fluorescence PCR was used to detect ureA gene of HP in feces from 50 patients,including 23 confirmed by gastric biopsy urease test and histological staining.Bacterium culture and serum antibody detection were also performed, and chi-square test was used to compare the sensitivity,specificity,positive predictive value and negative predictive value among three methods.Results The sensitivity,specificity,positive predictive value and negative predictive value of fluorescence PCR were 1.00,0.96,96.O%and 100.0%,while those for HP culture were 0.78,1.00,100.0%and 84.0%,and thee for serum antibody detection Was 0.96,0.74,76.O%and 95.0%.There were significant differences in sensitivity and negative predictive value between PCR and bacterium culture (X2=5.60 and 4.44,P<0.05),and significant differences in specificity and positive predictive value between PCR and serum antibody detection(X2=5.28 and 4.08,P<0.05).Conclusion ureA gene detection in feces by fluorescence PCR is of value for the diagnosis of HP infection.

Establishment of mice pneumonia model with imipenem-resistant Acinetobacter baumannii / 中华检验医学杂志

Objective To construct the mice pneumonia model with imipenem-resistant Acinetobacter baumannii and provide experimental model in anti-pan-resistant Acinetobacter baumannii therapy study. Methods A total number of 120 4-week-old BALB/C male mice were randomly selected and divided into three groups including micro-intratracheal injection, ultrasonic atomizing and nasal dripping. The mice were treated with methotrexate to induce hypo-immunity in every group. These BALB/C mice of normal immunity and hypo-immunity were infected through Imipenem-resistant Acinetobacter baumannii by microintratracheal injection, ultrasonic atomizing and nasal dripping, respectively. The morbidity, mortality,bacterial clearance rate and histopathology in lung were determined. Results The morbidities of BALB/C mice with hypo-immunity infected by micro-intratracheal injection and ultrasonic atomizing achieved 100%(30/30), while the mortalities were 100% (10/10) and 33.3% (3/10), respectively. Mice in two groups above displayed the influx of neutrophils, lymphocytes and macrophages in the peri-bronchial and alveolar interstitial space 12-24 h after pulmonary infection. In addtion, the mice in micro-intratracheal injection group displayed coUapse of partial alveolar walls, formation of abscesses and bacterial colonies in alveoli. While the lung pathology in mice of ultrasonic atomizing group was characterized by cell degeneration in some regions in the lungs, slight relaxation, congestion in alveolar wall vessels and normal of bronchial and alveolar tissue 24 h after inoculation. Degeneration in peri-tracheal and peri-bronchial areas was observed 24-48 h after inoculation, along with highly expanded pulmonary blood vessels and edems. The inflammation was reduced at 48 hours. There was no obvious pulmonary infection in BALB/C mice with hypo-immunity by nasal dripping with mortality of 0% (0/10) and no significant histopathologic change in lungs. Conclusions BALB/C mice with hypo-immunity pneumonia model with Imipenem-resistant Acinetobacter baumannii can be conducted by micro-intratracheal injection or ultrasonic atomizing, but the latter has the advantages of high-productivity, easy-operation, low-cost, time-saving and usefulness. Mice with normal immunity are not susceptible to imipenem-resistant Acinetobacter baumannii.

Antimicrobial resistanee and resistant genes of Pseudomonas aeruginosa / 中华临床感染病杂志

Objective To investigate the trend of antimicrobial resistance and the prevalence of resistant genes in Pseudomonas aeruginosa strains isolated from Hangzhou First People's Hospital.Methods Antimicrobial susceptibilities of 1489 Pseudomonas aeruginosa strains isolated from 2003 to 2007 were statistically analyzed using WHONET.MICs of 11 antimicrobisis to 36 multi-drug-resistant Pseudomonas aerugionosa strains were determined by agar dilution method.Genes of β-lactamases(BLA)and aminoglycoside-modifying enzymes(AMEs)were detected by PCR and the PCR products were sequenced.Results The resistant rates to aztreonam,imipenem,ceftazidime,cefepime,piperacillin,piperacillin/tazobactam.cefoperazone/sulbactam,ciprofloxacin,levofloxacin,gentamicin and amikacin were increased from 13.4%,10.6%,8.7%,7.9%,12.7%,12.7%,6.7%, 15.8%,20.5%,24.7% and 10.9%in 2003 to 35.3%,40.9%,18.4%, 32.4%,32.9%,32.0%,21.9%,37.8%,38.6%, 39.4% and 34.8% in 2007.respectively.Hish MICs of 11 antimicrobiMs for multi-drug resistant Pseudomonas aerouginosa were determined with MIC90≥128 μg/mL.In 36 multi-drug resistant Pseudomonas aeruginosa strains,21(58.3%)strains carried β-lactamase genes and 32 strains(88.9%)carried aminoglycosidemodifying enzyme genes,while the deletion rate of oprD2 was 80.6%(29/36).Conclusions The resistant rates to common antibiotics of Pseudomonas aeruginosa have increased.resulting in multi-drug resistance.Genes of β-lactamases and aminoglycoside-medifying enzymes are prevalent in multi-drug resistant Pseudomonas aeruginosa strains,with the common deletion of oprD2.

Antimicrobial Agents Resistance and Encoding Genes of Acinetobacter baumannii Isolated from Four Hospitals in Hangzhou City / 中华医院感染学杂志

OBJECTIVE To investigate the resistance and the distribution of the main ?-lactamases encoding gene in Acinetobacter baumannii isolated from four hospitals in Hangzhou city to provide the basic data for the optional treatment of A.baumannii infection.METHODS The identification of A.baumannii was performed using VITEK-AMS60.The minimum inhibitory concentrations(MIC) was examined by agar dilution and E-test.The homology of the resistant isolates was finished by pulsed field gel electrophoresis(PFGE).PCR and sequencing were used to analyze the ?-lactamases encoding gene of the 36 strains of imipenem-resistant A.baumannii.RESULTS All of the imipenem-resistant isolates produced carbapenemase OXA-23,and 5 isolates produced PER-1,2 isolates produced TEM-1 except OXA-23.No metallo-?-lactamases were detected.No plasmid was extracted.Clone transmission of the imipenem-resistant strains existed in the 4 hospitals.Most strains were isolated from intensive care unit(ICU). CONCLUSIONS The clone transmission of imipenem-resistant A.baumannii strains is occurred in 4 hospitals.All strains produce carbapenemase OXA-23.Five strains also produce PER-1 type extended-spectrum ?-lactamases.Two strains also produce TEM-1 type extended-spectrum ?-lactamases.

Drug Resistance of Alcaligenes xylosoxidans / 中华医院感染学杂志

OBJECTIVE To investigate the distribution and the antimicrobial resistance of Alcaligenes xylosoxidans and to provide the guidance of rational use of antibiotics.METHODS A.xylosoxidans was isolated and identified through VITEK-AMS all-automatic microbiology analysis system and its G~-bacilli identification cards(GNI) and drug sensitivity cards(GNS-KI/121) were also analyzed.Drug-resistance was tested by Kirby-Bauer disk(sensitivity) method((Bio-Merieux) and Oxoid products).RESULTS The drug-resistance rates to the 1st to 4th((except) CAZ) generation cephalosporins were more than 77%.The drug-resistance rate to the aminoglycosides was more than 76%,and to(ampicillin,) amoxicillin/CA,ampicillin/sulbactam,cefoxitin and cefuroxime were all 100%;but to(aztreonam) was 92.86%;66.67% and 44.44% strains were resistant to ciprofloxacin and(levofloxacin).(CONCLUSIONS) The detection rates of the multidrug resistant A.xylosoxidans have the tendency to increase yearly.The antibiotics should be used reasonably according to the results of drug sensitivity test in clinic.

Molecular Mechanisms of Macrolide Resistance of Mycoplasma pneumoniae / 中华医院感染学杂志

OBJECTIVE To investigate macrolide resistance and main molecular mechanisms in Mycoplasma pneumoniae. METHODS Thirty two throat swabs from children infected with M. pneumoniae were cultured by modified Hayflick medium. Antibiotic susceptibility test was used to screen the macrolide-resistant M. pneumoniae. The 23S rRNA gene sequences of the strains were determined with polymerase chain reaction and sequencing. RESULTS Nineteen strains were isolated from 32 throat swabs successfully.Fifteen strains were resistant to macrolide antibiotics according to the results of antibiotic susceptibility test. Once the strain was resistant to one of macrolide antibiotics,it would be resistant to the others. Sequencing results of the sensitive strains and the standard strain FH were completely same. Fifteen resistant strains presented A2063G point mutation in 23 SrRNA region Ⅴ, in which 2 examples showed the coexistence of the sensitive strain and the resistant strain. CONCLUSIONS Macrolide-resistant M. pneumoniae is common and serious at present. The antibiotic resistant isolate carries point mutations of the 23S rRNA region Ⅴ.

Surveillance of Pathogens Distribution and Antibiotic Resistance in Lower Respiratory Tract Infection in ICU / 中华医院感染学杂志

OBJECTIVE To investigate the distribution of pathogens and their antibiotic resistance in lower respiratory tract infection(LRTI) from intensive care unit(ICU) in our hospital,and provide basis for rational selection of clinical drugs.METHODS Pathogens were detected from qualified sputum specimens in LRTI from ICU and identified by VITEK-AMS60 automatic microbial analyzing system.Drug susceptibility was determined by KB test.RESULTS From 320 sputum specimens 367 pathogens were detected between from Jan 2007 to Mar 2008,including 261 strains(71.1%) of Gram-negative bacilli,70 strains(19.1%) of fungi,and 36 strains(9.8%) of Gram-positive cocci.21.8% Of the isolated pathogens were Acinetobacter baumannii,with 16.7% of drug-resistant rate to cefoperazone/sulbactam and over 71% to other 13 antibiotic agents.The rate of extended spectrum ?-lactamases(ESBLs) producing Escherichia coli isolates and Klebsiella pneumoniae ones were 65.2% and 72.0%,respectively,comparing to 84.6% for meticillin-resistant Staphylococcus aureus(MRSA).CONCLUSIONS Gram-negative bacilli are the major pathogens of LRTI in ICU,in which A.baumannii shows with a high rate of drug-resistance,followed by fungi,which should attract the clinician′s more attention.

Study on genic expression of ampC in Serratia marcescens / 中华检验医学杂志

Objectives To investigate the properties and distributions of ampC gene among different drug-resistant strains of Serratia marcescens,and the relationship of control gene ampR with AmpC enzymes′ expressions.Methods According to the results of inducting experiment with 1/2 MIC of beta-lactam antibiotics (CTX),three-dimensional testing and isoelectric focusing electrophoresis testing,143 strains of S.marcescens were classified into three groups:including induction group, continuous low-production group and hyperproduction group. In each group, the sequences of ampC and ampR genes were amplified using the method of PCR. The products of PCR were analyzed. The plasmid-mediated beta-lactamases were detected using the method of conjugation experiment.Results Among 143 strains of S.marcescens, the continuous low -production strains, induction strains and hyperproduction strains were 14,103,and 18, respectively.125 and 99 strains were ampC and ampR gene positive, respectively.The detection rate of ampR in hyperproduction group was lower than other groups.5 sites of ampC genes and 4 sites in the Open Reading Frame (ORF) of ampR gene were easily mutated in 5 induction strains and 2 hyperproduction strains.Conclusions The production of inducing drug-resistance of some S.marcescens might be related to mutation of ampC gene encoding AmpC beta-lactamases and the ORF mutation in ampR. The continuous hyperproduction drug-resistance had something to do with deletion mutation in ampR in segmental hyperproduction strains.The plasmid-mediated AmpC enzymes hadn′t been found in S.marcescens.

Research on Solar-UV Individual Exposure of Pupils in Shenyang / 环境与健康杂志

Objective To explore individual exposure to solar-UV radiation of pupils in Shenyang in four seasons. Methods Personal exposure to solar-UV radiation of pupils in Shenyang in four seasons was measured with ultraviolet radiation individual monitors and at the same time UV intensity in environment was detected also. Results Results of the present paper showed that higher UV exposure dosage of the pupils had been determined in spring and autumn, lower dosage had been determined in winter and summer respectively. The accumulative UV exposure time of pupils showed lower levels in winter compared with those in spring, summer and autumn. Higher UV exposure dosage and exposure time had been determined in class day than those in break day. The annual mean of daily UV exposure dosage and exposure time presented higer levels at noon. The mean of daily exposure of pupils was less than 8 percent of environmental UV intensity. Conclusion The distribution of UV exposure of pupils in daytime is corresponded with time spent outdoors. Individual differences are mostly caused by personal behavior. Seasons may affect personal UV radiation exposure. The UV exposure received by individuals only take a little part of total environmental UV radiation.