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OBJECTIVE@#Antimicrobial resistance (AMR) has become a global concern and is especially severe in China. To effectively and reliably provide AMR data, we developed a new high-throughput real-time PCR assay based on microfluidic dynamic technology, and screened multiple AMR genes in broiler fecal samples.@*METHODS@#A high-throughput real-time PCR system with an new designed integrated fluidic circuit assay were performed AMR gene detection. A total of 273 broiler fecal samples collected from two geographically separated farms were screened AMR genes.@*RESULTS@#The new assay with limits of detection ranging from 40.9 to 8,000 copies/reaction. The sensitivity rate, specificity rate, positive predictive value, negative predictive value and correct indices were 99.30%, 98.08%, 95.31%, 99.79%, and 0.9755, respectively. Utilizing this assay, we demonstrate that AMR genes are widely spread, with positive detection rates ranging from 0 to 97.07% in 273 broiler fecal samples. blaCTX-M, blaTEM, mcr-1, fexA, cfr, optrA, and intI1 showed over 80% prevalence. The dissemination of AMR genes was distinct between the two farms.@*CONCLUSION@#We successfully established a new high-throughput real-time PCR assay applicable to AMR gene surveillance from fecal samples. The widespread existence of AMR genes detected in broiler farms highlights the current and severe problem of AMR.
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Objective To study the incidence of stroke among elderly people in China's longevity area and its association with diseases such as hypertension,diabetes and heart disease.The differences in the following common hematological indicators in subjects with stroke and non-hypertension,diabetes,heart disease and stroke were studied:superoxide dismutase (SOD),malondialdehyde (MDA),hypersensitive c-reactive protein (hsCRP),albumin (propagated) glucose (GLU),cholesterol (CHO),triglyceride (TG),high-density lipoprotein cholesterol (HDLC),glycosylated serum protein (GSP) urea nitrogen (BUN),creatinine (CREA) and uric acid (UA).Methods Residents who participated in the project of biomedical research of aging population conducted in 2014 were selected from 8 longevity Areas in China.2 315 people aged 40 and over attended the study,including 22 aged 40 and over,238 aged 60 and over,490 aged 70 and over,629 aged 80 and over,518 aged 90 and over,418 aged 100 and over.Using the self-designed questionnaire to collect information about the characteristics of social demographics,the clinical doctors used the unified inspection tool to examine the subjects.The fasting blood samples were collected by vacuum tube at early morning.The contents of plasma SOD,MDA,hsCRP,ALB,GLU,CHO,TG,HDLC,GSP,BUN,CREA and UA were detected and compared among these elderly who were classified into different genders and different age groups and different healthy groups.Results The prevalence of high blood pressure,diabetes,heart disease and stroke increased with age,reaching a peak and then slowly decreasing.The age of peak was 90 ～ 99,60 ～ 69,70～ 79 and 80～ 89.The prevalence of hypertension was 71.62 ％ and 60.54 ％ respectively for stroke subjects and non-cerebral apoplexy subjects,and the difference was statistically significant.The prevalence of diabetes was 18.92％ and 11.35％ respectively,and the difference was statistically significant.The prevalence of heart disease was 20.98％ and 5.26％,respectively,and the difference was statistically significant.The rates of non-hypertension,non diabetic and non-heart disease were 4.73％ and 33.41％ respectively,and the difference was statistically significant.In the groups of Stroke subjects and Non-high blood pressure,nomdiabetic,non-heart disease subjects the following indicators were Compared,values of SOD were 55.76±8.27 and 57.16±8.00 U/ml respectively,the difference between groups were not statistically significant (t=0.341,P=0.053),values of MDA were 5.81 ± 3.82 and 5.67± 3.16 μmol/L respectively,the difference between groups were not statistically significant (t=0.329,P =0.661),values of hsCRP were 4.15 ± 12.33 and 2.94 ± 6.25 mg/L,respectively,the difference between groups were not statistically significant (t=0.026,P=0.080),values of ALB were 41.60±4.51 and 42.08±3.94 g/L respectively,the difference between groups were not statistically significant (t=0.032,P=0.194),values of ALB were 41.60 ± 4.51 and 42.08± 3.94 g/L respectively,the difference between groups were not statistically significant (t =0.032,P=0.194),values of ALB were 41.60 ± 4.51 and 42.08 ± 3.94 g/L respectively,the difference between groups were not statistically significant (t =0.032,P=0.194),values of GLU were 5.89 ± 2.67 and 4.90 ± 0.90 mmol/L respectively,the difference between groups showed statistically significant (t=0.000,P=0.000)),values of CHO were 4.81 ± 1.00 and 4.71±1.02 mmol/L respectively,the difference between groups were not statistically significant (t =0.670,P=0.318),values of TG were1.33±0.69 and 1.14±0.57 mmol/L respectively,the difference between groups showed statistically significant (t=0.012,P=0.000),values of HDLC were 1.29±0.35 and 1.41±0.40 mmol/L respectively,the difference between groups showed statistically significant (t=0.004,P=0.001),values of GSP were 259.10±60.90 and 246.75±24.52 μmol/L respectively,the difference between groups showed statistically significant (t =0.000,P =0.000),values of BUN were 6.84±± 3.53 and 6.62 ± 2.20 mmol/L respectively,the difference between groups were not statistically significant (t=0.110,P=0.338),values of CREA were 84.92 ± 33.00 and 80.14 ± 24.64 μmol/L respectively,the difference between groups showed statistically significant (t=0.013,P=0.044),values of UA were 296.73±91.34 and 288.12±80.47 μmol/L respectively,the difference between groups were not statistically significant (t=0.123,P=0.247).Conclusion Diabetes,hypertension,and heart disease are risk factors for stroke.Abnormal blood glucose and lipid metabolism:the increase of GLU,TG and the decrease of HDLC are important common biochemical index of stroke.Patients with cerebral apoplexy have certain renal impairment.
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Objective To investigate children with nephrotic syndrome (NS) receiving long-term glucocorticoid treatment while monitoring insulin resistance(IR) index,and analyze the influencing factors,and to observe the efficacy of metformin for preventing glucocorticoid-induced obesity and diabetes mellitus type 2.Methods A total of 32 cases including 20 boys and 12 girls with mean age of 7.6 years old(2 to 14 years old),were diagnosed as NS.All children undergoing long-term treatment of prednisone ＞ 1 mg/(kg · d) for at least 3 months.The height,body weight and body mass index(BMI) were monitored.The concentrations of serum insulin and C-peptide were determined by radioimmunoassay,the concentrations of serum HbA1 c were determined by high performance liquid chromatography,and the levels of fasting glucose,cholesterol and triglyceride were measured with an automatic measuring analyzer (American ABBO TT CCX-Ⅱ).The homeostasis model assessment-estimated insulin resistance(HOMA-IR) was calculated;When HOMA-IR ＞ 3.5,the patients were given metformin treatment for 12 weeks while supervising their height,weight,fasting blood glucose levels monthly.Results Out of 32 cases of NS with long-term use of prednisone,23 cases had HOMAIR ＞ 3.5 (72％).Thus,IR group included 23 patients,and non-IR (NIR) group had 9 patients.There were significant differences between 2 groups in BMI,age,prednisone dosage and the levels of triglyceride (TG) (all P ＜ 0.05).After 12 weeks of metformin treatment,the HOMA-IR (5.24 ± 1.82 vs 2.54 ± 1.09,P ＜ 0.01),HbA1 c levels [(6.36 ±0.82)％ vs(5.39 ±0.51)％,P ＜0.05],BMI[(27.42 ±6.12) kg/m2 vs(22.72 ±5.48) kg/m2,P ＜0.01],and TG levels[(2.03 ± 1.10) mmol/L vs(1.45 ±0.48) mmol/L,P＜0.05] were significantly reduced.Conclusions Older children with long-term use of glucocorticoid may easily lead to IR.Therefore,monitoring the HOMA-IR and early administration of mefformin is helpful for prevention and treatment of glucocorticoid-induced obesity and diabetes mellitus type 2.
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<p><b>OBJECTIVE</b>To establish and compare the pulsed-field gel electrophoresis (PFGE), multiple-locus variable number tandem repeat analysis (MLVA) and automated ribotyping for subtyping of Citrobacter strains.</p><p><b>METHODS</b>PFGE protocol was optimized in terms of plug preparation procedure, restriction enzymes and configuration of electrophoretic parameters. MLVA method was evaluated by finding variable number tandem repeats in two genomes of Citrobacter strains. The ribotyping was performed by using the automated RiboPrinter system.</p><p><b>RESULTS</b>We optimized the plug preparation procedure, focused on the cell suspension concentration (turbidity of 2.5 to 3.5), SDS addition (no SDS needed) and lysis time (1 h), and selected the appropriate restriction enzyme (XbaI) and the electrophoretic parameters (1.0 s-20.0 s for 19 h) of PFGE. There was nearly no discriminatory power of MLVA between Citrobacter strains. For 51 Citrobacter strains, automated ribotyping gave a D-value of 0.9945, while PFGE gave a D-value of 0.9969. Both PFGE and automated ribotyping clustered strains from the same sources (with the same species from the same place at the same time identified as the same source) and divided strains from different sources (from different years, places and hosts) into different subtypes.</p><p><b>CONCLUSION</b>PFGE protocol established in this paper and automated ribotyping are suitable for application in Citrobacter subtyping.</p>
Subject(s)
Automation , Citrobacter , Classification , Genetics , Electrophoresis, Gel, Pulsed-Field , Methods , Minisatellite Repeats , Genetics , Multilocus Sequence Typing , Methods , Phylogeny , Ribotyping , MethodsABSTRACT
Objective To develop a PFGE protocol for Streptococcus suis.Methods We developed and optimized a PFGE protocol for S.suis,in terms of plug preparation,choice of restriction endonucleases and optimized electrophoresis parameters.By analyzing the genome sequences of S.suis P1/7 with Mapdraw of DNAStar.we found three restriction enzymes,Swa Ⅰ,Sma Ⅰ and Apa Ⅰ,were more suitable than others.Results Analysis of 100 isolates of S.suis including 34 of 35 serotypes identified,59,53 and 43 patterns were obtained from Swa Ⅰ,Sma Ⅰ and Apa Ⅰ restriction,respectively.The enzyme Swa Ⅰ had the greatest power for discrimination ability.Conclusion By optimization of the protocol at various conditions,a rapid,reproducible,economic and practical PFGE method for S.suis was developed.