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【Objective】 To explore the distribution of polymorphisms of miR-208 genes rs8022522 and rs12894524 locus in Guangxi healthy population and compare the differences in the polymorphism distribution in different population. 【Methods】 SNPscan technology was used to detect genotypes of rs8022522 and rs12894524 from 297 healthy people in Guangxi, and the results were compared with other populations from Human genome Haplotype Map(HapMap) data. 【Results】 Three genotypes, namely, AA (2.7%), AG (24.2%) and GG (73.1%), in rs8022522 were found, with the allele frequencies of A and G being 14.8% and 85.2%. The genotypes of rs12894524 locus were TT (1.3%), TG (13.5%) and GG (85.2%), and the frequency of T and G allele was 8.1% and 91.9%, respectively. rs8022522 and rs12894524 locus genotypes and allele frequencies were significantly different from HapMap-CEU, HapMap- YRI and HapMap-TSI (P<0.05). Compared with HapMap-JPT and HapMap-CHB, there was no significant difference in genotype or allele frequency between the two sites (P>0.05). As for the blood lipid level among the three genotypes in rs8022522, the level of high density lipoprotein cholesterol (HDL-C) with GG genotype was significantly different from that in AG group (P<0.05). 【Conclusion】 The polymorphisms of rs8022522 and rs12894524 of miR-208 gene in Guangxi population are different from those in other regions to varying degrees. The polymorphism of rs8022522 locus is related to the level of HDL-C.
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【Objective】 To investigate the situation of hepatitis E virus(HEV) infection among voluntary blood donors in Wuhan area and provide evidences for enhancing blood screening strategies. 【Methods】 HEV nucleic acid detection(NAT) was performed on blood samples from eligible blood donors in Wuhan from November to December 2020. The testing results were analyzed, and the blood donors with repeated reactive results were followed up to clarify the status of infection. 【Results】 Routine screening was performed on 17 409 blood samples from November to December 2020. A total of 17 322 blood samples of eligible blood donors were tested for HEV NAT, and one case of HEV RNA reactivity was detected. The results from the follow-ups showed that the blood donor should be in the window period of HEV seroconversion. The current HEV infection rate of voluntary blood donors in Wuhan arewas 0.058‰(1/17 322), which was lower than other domestic areas. 【Conclusion】 The current HEV infection rate of voluntary blood donors was at a relatively low prevalence level in Wuhan area. Selective blood screening strategies can be taken to further reduce potential risk of blood transfusion infection with hepatitis E virus.
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Objective:To observe the application value of dizocine sedation in brachial plexus nerve block.Methods:From January 2016 to December 2018, 80 patients with brachial plexus block anesthesia in the Second People's Hospital of Tongxiang were selected and divided into two groups according to the random number table method, with 40 cases in each group.The control group was not given sedation intervention, while the observation group was given dizocine sedation.The heart rate, mean arterial pressure, blood oxygen saturation and Ramsay scores at different time points were compared between the two groups.The anesthetic effect of the two groups was analyzed.Results:The onset time of sensory block in the observation group[(8.86±0.92)min] was significantly shorter than that in the control group[(11.43±1.08)min]( t=11.457, P<0.05). The duration of sensory block[(729.95±54.43)min] and the duration of analgesia[(879.96±165.52)min] in the observation group were significantly longer than those in the control group[(600.73±49.86)min, (750.27±261.19)min]( t=11.072, 2.653, all P<0.05). There were no statistically significant differences in the mean arterial pressure, heart rate and blood oxygen saturation at each time point between the two groups before operation (all P>0.05). The mean arterial pressure and heart rate of the observation group were significantly lower than those of the control group at 15 min, 30 min after operation and at the end of operation (all P<0.05). The Ramsay score between the two groups had no statistically significant difference before operation ( P>0.05). At 15 min, 30 min after operation and at the end of surgery, the Ramsay scores of the observation group were (4.08±0.54)points, (4.15±0.37)points, (2.96±0.19)points, respectively, which were significantly higher than those of the control group [(1.79±0.27)points, (1.77±0.16)points, (1.93±0.14)points], the differences were statistically significant ( t=23.989, 37.341, 27.602, all P<0.05). Conclusion:After dizocine sedation is used in brachial plexus block patients, the vital signs fluctuate slightly and the sedation effect is obvious.
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Objective To understand the prevalence of Salmonella and the characteristics of drug resistance genes in General Hospital of People's Liberation Army, and to provide evidence for the prevention and treatment of Salmonella infection. Methods A retrospective study was conducted to collect 78 clinical isolates of Salmonella from 2015 to 2017. The age of the patients was 49 ± 21 years old. The infected patients were mainly young and middle-aged. The clinical samples mainly came from feces and venous blood, accounting for 44.87%(35/78) and 33.33%(26/78), respectively. After serotype identification, drug sensitivity test and whole genome sequencing, multilocus sequence typing and drug resistance genotyping were performed. Cluster of Cefotaxime or Ciprofloxacin resistant Salmonella was analyzed. Results Salmonella group D (53.85%) and Salmonella group C (21.79%) were dominant Salmonella serotype. ST11 was mainly ST type. Drug sensitivity test showed that the multidrug resistance rate of Salmonella was 64.11% (50/78). The sensitivity to all antimicrobial agents' rate was 25.64 (20/78). The resistance rate of Salmonella to nalidixic acid was 65.38%(51/78). The most common drug resistance gene of Salmonella was extended-spectrum β-lactam drug resistance gene, accounting for 78.21% (61/78). Conclusions The ST-type and carrying resistance genes of Salmonella in this hospital were diverse. Most pathogens were multi-drug resistant to antimicrobial agents. Molecular typing and drug resistance gene analysis of Salmonella and construction of resistant strains to determine the inheritance of Salmonella relationships have a certain clinical significance.
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Objective@#To discuss the application of matrix assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF MS) in the identification of Aspergillus and evaluate its performance.@*Methods@#the clinical isolates of Aspergillus collected from May 2017 to March 2018 in PLA General Hospital were identified by VITEK MS V3.0 and the results were analyzed. The ITS sequencing resultswere used as the gold standard.@*Results@#It identified 9 Aspergillus species (including 12 Aspergillus species in total) through the V3.0 database, accounting for 86.24% of the total clinical isolates. The identification rate by VITEK MS was 91.49% with 16.51% was not identified. The coincidence rate of genus was 93.62%, of which only two Aspergillus versicolor were identified to the level of the genus. According to the confidence level analysis, 88.30% of the strains obtained more than 99% of the identification rate. 13.83% of the strains did not have the identification results for the first time, with the error rate of 3.19%. After secondary extractions, the percentage of unidentified strain was reduced to 6.38%, and the identification error rate was reduced to 2.13%. Combined with traditional identification and VITEK MS identification, the correct rate of strains identification was 98.94% on genus level, and was 93.62% on species level. The influence of other fungi on Aspergillus identification was 0%.@*Conclusion@#As a powerful supplement to the traditional identification method, MALDI-TOF MS showed a lot of convenience when applied in the identification of Aspergillus, which improves the identification accuracy and the identification ability for fungi in laboratory.(Chin J Lab Med, 2018, 41: 577-582)
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Objective To investigate the spectrum and antimicrobial resistance of major pathogens causing nosocomial infections in China, 2016. Methods Non-duplicated nosocomial cases as well as pathogens causing bloodstream infections ( BSI) , hospital-acquired pneumonia ( HAP) and intra-abdominal infections ( IAI ) from 12 teaching hospitals across China were collected. The minimum inhibitory concentrations (MICs) of important clinical common strains were determined by agar dilution method or broth microdilution method. The CLSI M100-S27 criteria was used for interpretation. Data were analyzed by using WHONET-5. 6 software. Results A total of 2060 cases were collected, including 894 cases from BSI, 630 cases from HAP and 536 cases from IAI. The MICs of 1896 important clinical common strains were determined. Escherichia coli and Klebsiella pneumoniae were the most prevalent pathogens causing BSI and IAI, while Acinetobacter baumanii and Pseudomonas aeruginosa were dominated in HAP. All Staphylococcus aureus were susceptible to tigecycline, linezolid, daptomycin and glycopeptides. Methicillin-resistant S. aureus accounted for 44. 4% ( 75/169 ) of all the S. aureus. The rate of methicillin-resistant coagulase-negative staphylococci was 80. 9% ( 72/89 ) . No Enterococcus strains were found resistant to tigecycline, linezolid or daptomycin. Vacomycin resistant enterococcus was found in Enterococcus faecium, accounting for 1. 8% ( 2/111 ) of all E. faecium strains. Tigecycline, meropenem, amikacin, imipenem, and polymyxin B exhibited high potency against Enterobacteriaceae and the susceptibility rates were 96. 6%(865/895), 94. 3% (859/911), 94. 2% (858/911), 94. 1% (857/911), and 91. 6% (820/895), respectively. The prevalence of extended-spectrum β-lactamase was 58. 4% ( 263/450 ) in E. coli and 28. 6% ( 84/294 ) in K. pneumonia. The rate of carbapenem resistant K. pneumonia and E. coli was 15. 3% ( 45/294 ) and 1. 8% ( 8/450 ) , respectively. The percentage of polymyxin B resistant K. pneumonia and E. coli was 4. 1% ( 12/294 ) and 4. 4% ( 20/450 ) , respectively. The rate of tigecycline resistant K. pneumonia and E. coli was 2. 4% ( 7/294 ) and 0. 2% ( 1/450 ) , respectively. A. baumanii showed low susceptibility to the antimicrobial agents except tigecycline ( 91. 4%, 235/257 ) and polymyxin B (100%, 257/257). The rate of carbapenem resistant A. baumanii was 80. 5% (207/257). The rate of carbapenem resistant P. aeruginosa was 31. 7% ( 59/186 ) . Polymyxin B and amikacin demonstrated high antibacterial activity against P. aeruginosa with susceptility rate of 100% ( 186/186 ) and 90. 9% ( 169/186), respectively. Conclusions Nosocomial pathogens showed high susceptibilities against tigecycline and polymyxin B. Antimicrobial resistance in A. baumannii is a serious problem. The prevalence of carbapenem-resistant Enterobacteriaceae and polymyxin B resistant Enterobacteriaceae has increased, which should be monitored continuously in China.
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To dynamically investigate the distribution and antimicrobial resistance profiles of bacteremia pathogens isolated from different regions in China in 2011, 2013 and 2016. Non-repetitive isolates from nosocomial bloodstream infections were retrospectively collected and detected for antimicrobial susceptibility tests (AST) by agar dilution or microbroth dilution methods. Whonet 5.6 was used to analyze the AST data. Among 2 248 isolates, 1 657 (73.7%) were Gram-negative bacilli and 591 (26.3%) were Gram-positive cocci. The top five bacteremia pathogens were as follows, Escherichia coli (32.6%, 733/2 248), Klebsiella pneumoniae (14.5%, 327/2 248), Staphylococcus aureus (10.0%, 225/2 248), Acinetobacter baumannii (8.7%, 196/2 248) and Pseudomonas aeruginosa (6.2%, 140/2 248). Colistin (96.5%, 1 525/1 581, excluding innate resistant organisms), tigecycline (95.6%, 1 375/1 438, excluding innate resistant organisms), ceftazidine/clavulanate acid (89.2%, 1 112 /1 246), amikacin (86.4%, 1 382/1 599) and meropenem (85.7%, 1 376/1 605) showed relatively high susceptibility against Gram-negative bacilli. While tigecycline, teicoplanin and daptomycin (the susceptibility rates were 100.0%), vancomycin and linezolid (the susceptibility rates were 99.7%) demonstrated high susceptibility against Gram-positive cocci. The prevalence of extended-spectrum β-lactamases (ESBLs)-producing Enterobacteriaceae were 50.6% (206/407), 49.8% (136/273) and 38.9% (167/429) in 2011, 2013 and 2016 respectively; carbapenem-non-susceptible Enterobacteriaceae were 2.2% (9/408), 4.0% (16/402) and 3.9% (17/439) in 2011, 2013 and 2016 respectively; The prevalence of multidrug-resistant A. baumannii (MDRA) was 76.4% (55/72) in 2011, 82.7% (43/52) in 2013 and 87.5% (63/72) in 2016, respectively. The prevalence of multidrug-resistant P. aeruginosa (MDRP) was 9.8% (5/51) in 2011, 20.0% (7/35) in 2013 and 13.0% (7/54) in 2016, respectively. The prevalence of methicillin-resistant S. aureus (MRSA) was 51.9% (41/79) in 2011, 29.7% (19/64) in 2013 and 31.7% (26/82) in 2016, respectively. The prevalence of high level gentamicin resistance (HLGR) of Enterococcus faecium and Enterococcus faecalis were 43.2% (48/111) and 40.9% (27/66), respectively. The predominant organism of carbapenem-non-susceptible Enterobacteriaceae was K. pneumoniae with its proportion of 57.1% (24/42). Among 30 tigecycline-non-susceptible Enterobacteriaceae, K. pneumoniae was the most popular organism with 76.7% (23/30). Among 39 colistin-resistant Enterobacteriaceae, E. coli, Enterobacter cloacae and K. pneumoniae were constituted with the percent of 43.6 (17/39), 35.9 (14/39) and 15.4 (6/39), respectively. The Gram-negative bacilli (E. coli and K. pneumoniae were the major organisms) were the major pathogens of nosocomial bacteremia, to which tigecycline, colistin and carbapenems kept with highly in vitro susceptibility. Whereas, among the Gram-positive cocci, S. aureus was the top 1 isolated organism, followed by E. faecium, to which tigecycline, daptomycin, linezolid, vancomycin and teicoplanin kept with highly in vitro susceptibility. Isolation of colistin-resistant Enterobacteriaceae, tigecycline-non-susceptible Enterobacteriaceae, linezolid- or vancomycin-non-susceptible Gram-positive cocci suggests more attention should be paid to these resistant organisms and dynamic surveillance was essential.
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Objective To shorten the turn around time of positive blood culture results by optimizing the blood culture positive specimen processing flow.Methods In January 26,2015,the microbiology department started the blood culture positive specimen processing flow optimization project,and applied the Lean Six Sigma method in the microbiological process management.The TAT data of 124 positive blood cultures containing Enterobacteriaceae were collected before and after the start of the project in about two months.We analyzed the turnaround time median,mean and standard deviation and reference Z value,process performance index,millions of error opportunities.We decompose the turnaround time into six time periods to find the key points of the process improvement and the influencing factors,and then put forward the reform measures to optimize the blood culture inspection process.MiniTab17.0 statistical software was used to process capability analysis and double sample t test.Results After the implementation of the project,the average turnaround time of the blood culture was shortened from 77.10 h to 64.03 h,improved by 13.06 h(16.94%).Process performance greatly improved in Ppk value increased from 0.49 to 0.88,the benchmark Z value increased from 1.48 to 2.63.After the improvement,except the positive alarm time of blood culture,the mean of the other decomposition time was significantly shorter than before.Conclusions The application of Six Sigma in process management can greatly improve the work efficiency and process performance.This project can save a lot of manpower,material and financial resources,reduce the waiting,shorten turnaround time,that achieve the desired results.
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Objective To assess the efficacy and safety of morinidazole combined with appendectomy in treating purulent or gangrenous appendicitis.Methods Double-blind randomized controlled multicenter clinical trial was designed and conducted.Totally 437 patients were included,219 in the control group and 218 in the experimental group.Cases of purulent or gangrenous appendicitis were enrolled and assigned to each of the two groups.The control group received ornidazole injection for 5 to 7 days while the experimental group received morinidazole injection.Both groups underwent appendectomy.Clinical response,micrombiological outcomes,overall response were evaluated.Adverse events and side effects were recorded.Results No significant difference was observed between the two groups regarding the clinical healing rate at 5-10 days after medicine withdrawal,anaerobia clearance and overall healing rates.Adverse events occurred in 140 patients (32.1%).Incidence of adverse events in the control group and the experimental group was 34.7% and 29.4%,respectively (P > 0.05).The overall incidence of side effects was 15.1% (66 cases).Side effects were less seen in the experimental group compared with that in the control group (11.5% vs.18.7%,P < 0.05).The most frequent side effects were aminotransferase rising,thrombocytosis,nausea,vomiting and electrocardiographic abnormality.Conclusions The effect of morinidazole plus operation was comparable with ornidazole in treating purulent or gangrenous appendicitis.The safety of morinidazole is better than ornidazole.
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Aims Toevaluatethepharmacodynamic efficacy of different types of antiangiogenic agents as HM-3 on a non-small cell lung cancer xenografts tumor model .To explore the interaction between the antian-giogenic agents and the tumor microenvironment,and to offer suggestions for clinical therapy.Methods Thenon-smallcelllungcarcinomaxenograftmodelwas established in Balb/c nude mice.The model mice were treated with Docetaxel(10 mg·kg-1 )as the positive control.The mice were parallelly treated with,HM-3 at the doses of 3 mg · kg-1 and 48 mg · kg-1 and, Avastin(5 mg·kg-1 ).The parameters include tumor volume,tumor weight and immunohistochemical analy-sis.Result Animalexperimentsshowedthatdocetaxel had good anti-tumor activity.Tumor growth inhibition by tumor weight of G2 docetaxel(10 mg·kg-1 )group was 60. 80%.Tumor growth inhibition by tumor weight of G3 HM-3(3 mg·kg-1 )group,G4 HM-3(48 mg· kg-1 )group ,G4 Avastin(5 mg·kg-1 )group,were 43. 60%,-34. 80%,44. 40%,respectively.Con-clusion Theantigiogeniceffectisaffectedbytumor growth stage,tumor microenvironment and their work-ing mechanisms.Angiogenesis inhibitors HM-3 has a certain effect of inhibiting tumor growth,but to little a-vail.HM-3 shows on inhibitory effect in a dose-de-pendent manner at the doses of 0~6 mg·kg-1 .HM-3 at a high dose of 48 mg · kg-1 has no inhibitory but promoting effects on human non-small cell lung carci-noma A549 xenografts in nude mice .Special dose-effect relationship indicates that dosage should be paid attention to in the clinical use of blood vessel inhibi-tors.
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Objective To compare of the difference about ectopic activation between autogenous bone graft and allograft from large segment tibia of rabbits.Methods Eighty healthy adult Chinese rabbits (6 months of age),weighing (2.5 ±-0.5)kg,were randomly divided into experimental group (allogeneic bone group) and the control group (autograft group),40 rabbits in each group.Another 10 rabbits were allogeneic bone donor.In experimental group,when 1.5 cm long rabbit tibial allograft were finishied,they were implanted into spatium intermusculare between the musculus rectus femoris and medial vastus muscle of the rabbit around the saphenous artery and were fastened to the femur by 1.0 mm Kirschner-wire.In control group,autologous tibias were done,the same as experimental group including length and position and method.Four weeks and 8 weeks and 12 weeks postoperative,respectively,the postmortem specimens were examined gross and immunohistochemistry and the expression of BMP-2 and collagen type Ⅰ of transplanted bone tissue were detected.Results BMP-2 mainly exist in cytoplasm of osteoblasts and chondrocytes undifferentiated mesenchymal cells.Collagen type Ⅰ primarily exist in the bone matrix around the pit of bone.The expression level of BMP-2 of experimental group in postoperative 4,8,12 and 16 weeks were 85.25 ± 4.47,109.44 ± 14.69,141.85 ± 9.45,116.25 ± 14.18,respectively,and the expression level of BMP-2 of control group were 103.78 ±-6.59,124.95 ± 14.94,145.46 ± 8.10,112.48 ± 13.27,respectively.The expression level of collagen type Ⅰ of experimental group in postoperative 4,8,12 and 16 weeks were 78.74 ± 7.99,95.95 ± 6.99,139.91 ± 4.32,137.76 ± 3.48,respectively,and the expression level of BMP-2 of control group were 88.87 ± 11.26,102.45 ± 2.82,140.76 ± 4.62,139.05 ± 4.55.Compared with control group,there was a significant difference in the expression level of the BMP-2 and collagen type Ⅰ of experimental group in postoperative 4,8 weeks (P < 0.05),but,there was no significant difference in the expression level of the BMP-2 and collagen type Ⅰ of experimental group in postoperative 12,16 weeks (P > 0.05).The amount increased gradually during 4 weeks,8 weeks,12weeks,peaked at 12 weeks,BMP-2 displayed a downward trend at 16 weeks,and collagen type Ⅰ basiclly maintain the level of 12 weeks.Conclusion Allograft segment could complete activation while they are implanted into spatium intermusculare containing famous blood supply within 3 months,there is no significant difference between autologous bone and allograft,it shows the feasibility of ectopic activation about allograft segment.
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Klebsiella Pneumoniae is an important pathogen for various infections clinically,in which hypervirulent Klebsiella pneumoniae (hvKP) mainly causes pyogenic liver abscess.By now,there are no standardized method to identify hvKP strains.hvKP strains are usually with hypermucoviscous phenotype,and the prevailing serotypes and clonal types are K1,K2 and CC23,CC65,respectively.Genes rmpA,magA,alls and kfu contribute to the pathogenicity of hvKP.This paper reviews the identification,clonal types and serotypes,hypermucoviscous phenotype and some other virulent genes of hvKP.
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Objective To observe the damage degree and expression pattern of Caveolin-3 mRNA by ischemia-reperfusion injury in rabbits of skeletal muscle cell at different phases.Methods In this study,from April 2013 to December 2013,30 lower limbs of 15 Chinese White Rabbits were used and divided into two groups:all the left lower limbs were experimental group,which were made as an experimental model of ischemia-reperfusion injury by occluding left common iliac artery using noninvasive vascular.All the right lower limbs without surgical treatment were the control group.Gastrocnemius samples were obtained at 4h and 8h after reperfusion and handled by HE staining and observed by optical microscopy.By Real-time PCR,Caveolin-3/GAPDH mRNA were detected.Results HE stain showed:in control group,there was no edema,degeneration and inflammatory cell infiltration; in experi-meatal group,muscle cell degeneration had occured at ischemic 5 h.The edema was aggravated,a large number vacuole were formed and inflammatory cell were infiltrated at 4 h reperfusion.Reperfusion injury at 8h significantly reduced compared to 4 h.The Caveolin-3/GAPDH mRNA expression levels by SPSS 19.0 showed:Control group:1.026 ± 0.065,1.004 ±0.037,1.022 ±0.051,experimental group:1.159 ±0.073,1.445 ±0.053,1.208 ±0.058 at ischemic 5 h,4 h and 8 h reperfusion,respectively.On-line analysis of variance cases of ischemic 5 h and 4 h reperfusion and 8 h reperfusion,the experimental group than the control group were increased,with statistical significance (P < 0.05).The experimental group of ischemic 5 h and 8 h reperfusion was no significant difference (P > 0.05).It showed Caveolin-3 mRNA expression levels in ischemia-reperfusion 8 h group returned to normal.There was significant statistical difference between the ischemic 5 h and 4 h reperfusion (P < 0.05).There was significant statistical difference between the 4 h reperfusion and 8 h reperfusion (P < 0.05).Conclusion The expression of Caveolin-3 in experimental group showed a trend of first increased and then decreased.The expression levels of Caveolin-3 mRNA in skeletal muscle cells after ischemia-reperfusion injury is consistent with the development and progression of muscle cell damage.The results indicate that Caveolin-3 may play a control role in the injury and recovery of skeletal muscle cell.
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<p><b>OBJECTIVE</b>To analyze horizontal transmission patterns of Streptococcus mutans among caries-active preschool children for early interventions of dental caries.</p><p><b>METHODS</b>Plaque samples obtained from 20 caries-active preschool children between 4 and 5 years of age were cultured under anaerobic conditions for isolating S. mutans, which were identified by morphological and biochemical analyses and PCR using primers homologous to the surface protein glucosyltransferase B (gtfB). The genotypes of the isolated S. mutans strains were determined by arbitrarily primed PCR (AP-PCR).</p><p><b>RESULTS</b>Of the 200 S. mutans isolates obtained, 19 were excluded by biochemical analysis, and the remaining 181 isolates were identified as S. mutans by PCR with primers of gtfB, showing 37 different genotypes as identified by AP-PCR. Six children were found to carry S. mutans of a single genotype, 11 carried 2 genotypes, 2 had 3 genotypes, and 1 had 4 genotypes; 2 children from different classes were found to carry S. mutans of the same single genotype.</p><p><b>CONCLUSION</b>We identified 37 genotypes of S. mutans in these caries-active preschool children, among whom horizontal transmissions of the strains were not found.</p>
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Pré-Escolar , Humanos , Cárie Dentária , Microbiologia , Placa Dentária , Genótipo , Glucosiltransferases , Reação em Cadeia da Polimerase , Infecções Estreptocócicas , Streptococcus mutans , ClassificaçãoRESUMO
Objective To investigate the quinolone resistance determinants in ciprofloxacin-resistant Acinetobacter bau-mannii (ABA)clinical isolates.Methods One hundred and fourteen ciprofloxacin-resistant ABA strains were collected from six Chinese hospitals .The quinolone resistance determining region ( QRDR) of 4 target genes ( gyrA, gyrB, parC and parE) was amplified , sequenced and compared with the reference genome of ATCC 17978 to identify possible resistance-related mutations.Nine plasmid-mediated quinolone resistance (PMQR) genes (qnrA, qnrB, qnrC, qnrD, qnrS, qepA, aac(6′)-Ⅰb-cr, oqxA and oqxB) were also amplified, and the amplicons were then sequenced to determine their character-istics.Results Almost all isolates (113/114, 99.1%) harbored a substitution in codon 83 of gyrA gene, leading to a Ser83Leu mutation.Meanwhile,58.8%(67/114) of the isolates possessed dual mutations of GyrA-Ser83Leu and GyrA-Ser80Leu, which were known determinants for ciprofloxacin resistance .There were also multiple non-synonymous substitu-tions in gyrB, leading to Arg393Ser, Arg393Cys, Thr401Ala, Pro406Ser, Val430Phe, Cys440Ser and Gly480Arg muta-tions with prevalence rates of 95.6%, 0.9%, 96.5%, 96.5%, 100%, 96.5%and 96.5%,respectively.For parE, all the seven mutations were synonymous and found in more than 96%of the tested isolates.For PMQR genes, although 83.3%(95/114) of the isolates were positive for aac(6′)-Ⅰb, nocrmutations were identified.None of the other eight PMDR genes were found in our strain collection .Conclusion Although multiple mutations are identified in gyrB and parE, these mutations might be the characteristic SNP markers for specific clones , unlikely linked to quinolone resistance .No PMQR is found in the tested isolates.Mutations in chromosomal QRDR (GyrA-Ser83Leu and ParC-Ser80Leu) are the main determi-nants of ciprofloxacin resistance in our ABA collection .
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With the rapid development of clinical laboratory technology and clinical improvement,the discordance between department of microbiology and clinical departments got more and more.Based on the analysis of interface issues existed between them,we discussed and explored how to do a wonderful job to communicate with each other.(Chin J Lab Med,2013,36:375-376)
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<p><b>OBJECTIVE</b>To screen of high cariogenicity Streptococcus mutans (S. mutans) strains isolated from clinical specimens preliminary.</p><p><b>METHODS</b>Acidogenicity, aciduricity, extracellular polysaccharide production and adhesion of 41 strains of S. mutans isolated from clinical specimens were investigated to screen high cariogenicity S. mutans strains.</p><p><b>RESULTS</b>There were different cariogenicity among 41 strains of S. mutans, in which 3 strains of S. mutans had all high ability to produce extracellular polysaccharide, adhere to the saliva-coated hydroxyapatite, produce acid and tolerate acid, indicated there were 3 strains with high cariogenicity S. mutans strains isolated from clinical specimens. Another 3 strains of S. mutans with all low ability to produce extracellular polysaccharide, adhere to the saliva-coated hydroxyapatite, produce acid and tolerate acid indicated they were low cariogenicity S. mutans strains isolated from clinical specimens.</p><p><b>CONCLUSION</b>We may have obtained high cariogenicity S. mutans strains isolated from clinical specimens.</p>
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Humanos , Cárie Dentária , Durapatita , Saliva , Streptococcus mutansRESUMO
<p><b>OBJECTIVE</b>To identify Streptococcus mutans (S. mutans) strains from clinical samples.</p><p><b>METHODS</b>Plaque samples from caries-active and caries-free sites on enamel surfaces were obtained and cultivated for S. mutans isolation. Morphology, biochemistry, automatic microorganism analysis system and polymerase chain reaction using primers homologous to surface protein antigen I/II (spaP), glucosyltransferase B (gtfB) and dextranase (dexA) were used to identify S. mutans. Genotype of isolated S. mutans was determined by arbitrarily primed polymerase chain reaction.</p><p><b>RESULTS</b>Forty-six strains of S. mutans were obtained from the 32 subjects and were identified as S. mutans by biochemistry, automatic microorganism analysis system and polymerase chain reaction. Five identical genotypes were found by arbitrarily primed polymerase chain reaction.</p><p><b>CONCLUSION</b>Forty-one strains of S. mutans with different genotype were obtained from clinical samples.</p>
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Humanos , Cárie Dentária , Placa Dentária , Genótipo , Glucosiltransferases , Reação em Cadeia da Polimerase , Streptococcus mutansRESUMO
<p><b>OBJECTIVE</b>To select and identify ssDNA aptamers specific to Streptococcus mutans strains with different cariogenicity isolated from clinical specimens.</p><p><b>METHODS</b>Subtractive SELEX technology targeting the whole intact cells was used to screen for ssDNA aptamers specific to the clinical isolates Streptococcus mutans strains with different cariogenicity. Radioactive isotope, flow cytometry, gene cloning and sequencing, MEME online software and RNA structure analysis software were employed to analyze the first and secondary structures of the aptamers and identify the screened aptamers.</p><p><b>RESULTS</b>Detection by radioactive isotope showed sufficient pool enrichment after 9 rounds of subtractive SELEX. Flow cytometry showed that the selected aptamers H1, H16, H4, L1, L10 and H19 were capable of binding specifically with highly cariogenic Streptococcus mutans strains but not with strains with a low cariogenicity. The aptamer H19 had the strongest binding capacity to highly cariogenic Streptococcus mutans strains, with a dissociation constant of 69.45∓38.53 nmol/L.</p><p><b>CONCLUSION</b>We have obtained the ssDNA aptamers specific to the clinical isolates of highly cariogenic Streptococcus mutans strains.</p>
Assuntos
Humanos , Aptâmeros de Nucleotídeos , Genética , Clonagem Molecular , Primers do DNA , Cárie Dentária , Microbiologia , Biblioteca Gênica , Conformação de Ácido Nucleico , Técnica de Seleção de Aptâmeros , Especificidade da Espécie , Streptococcus mutans , Classificação , GenéticaRESUMO
Objective Exploring different strengthening immune strategy on long-term immune memory effects for tuberculosis vaccine lays the theoretical foundation.Methods The fusion protein AMM,adjuvant DDA and BCG-PSN mixed built AMM subunit vaccine.Choose only 50 C57BL/6 mice (SPF),according to the table of random number method was divided into 5 groups and each group 10 only,the experiment 1 group injected mice to phosphate buffer solution (PBS) as control; Experiment 2 mice only injected BCG vaccine (BCG) from early; Experiment 3 mice BCG initial free,in 10 weeks with AMM subunit vaccine strengthen immunity; The four mice from early after BCG,respectively on 8 weeks,10 weeks with AMM subunit vaccine strengthen immune mice one time.Experimental 5 groups of mice after BCG initial free,respectively on 6 weeks,8 weeks,10 weeks with AMM subunit vaccine strengthen immune mice a.28 weeks to remove spleen lymphocytes in mice,add 2.5μg/ml Ag85B stimulus 72 h after collecting spleen cell culture supernatant,with ELISA test Ag85B specificity splenic lymphocyte caused IFN ppar-gamma level.Results Spleen cells by antigen Ag85B stimulation,BCG initial free,AMM subunit vaccine strengthen immune once,twice,three times group and PBS compared all had difference (P <0.01 orP <0.05) ;BCG initial free-AMM subunit vaccine strengthen immune once,twice,three times group and BCG compared all had difference (P <0.01 or P <0.05) ; BCG initial free-AMM subunit vaccine strengthen immune once and twice or three times group compared with difference (P <0.01 orP <0.05) ; Initial free-AMM subunit vaccine strengthen immune two group and three times than group difference was statistically significant (P >0.05).Conclusion Mice strong immune strengthened the number is not the more the better,the appropriate number of strengthen the immune to induce the most lasting immunity memory,subunit vaccine strengthen immunization strategy may affect the immune memory is one of the important factors.