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Objective:To analyze the molecular epidemiological characteristics of Campylobacter fetus subsp. testudinum ( Cft). Methods:Fifteen strains of Cft collected in our laboratory from 2010 to 2022 were subjected to whole-genome sequencing. Their epidemiological characteristics were analyzed based on the global genome data of Cft on GenBank database. MLST-GrapeTree software was used to obtain the genetic structure of Cft strains. A phylogenetic tree was constructed using core-genome single nucleotide polymorphism (cgSNP) analysis, and the sequence clusters were identified using rhierBAPS. Virulence genes and drug resistance genes of Cft strains were annotated using CARD, ResFinder and VFDB database. Their susceptibility to antibiotics was tested using E-test method and the results were analyzed using the CLSI-M45 sensitivity standard for Campylobacter jejuni/ Campylobacter coli. Results:Based on average nucleotide identity (ANI) analysis, the genome data of 41 Cft strains including 24 isolated from human, 13 from animals and four of unknown sources were collected from GenBank database. Among the 24 human-derived strains, 20 were linked to Asian descent and only one was linked to Caucasian descent (spouse of Asian descent), showing statistically significant differences in human ethnicity. All of the 13 animal-derived strains were originated from reptilian sources, including six from turtles, four from snakes and three from lizards. MLST revealed that ST46 was the predominant ST in China, while ST15 was the major sequence type in the United States. Grapetree analysis also demonstrated that the genetic diversity in China was greater than that in the United States. The phylogenetic tree constructed based on cgSNP and BAPS identified six distinct sequence clusters. The Chinese isolates were scattered in diverse sequence clusters and closely related to animal-derived strains, while the American isolates mainly belonged to ST15. The genes encoding virulence factors such as flagella, glycosylation systems and adhesins were carried by all of the 41 Cft strains (100.00%). The invasion-related virulence genes, such as the genes encoding the IV type secretion system ( virB4, virB9, virD4) and the resistance-related tetO efflux pump gene were specifically identified in the emerging ST74 clones. In vitro drug susceptibility testing of 15 Chinese isolates revealed 46.67% of the Cft strains were resistant to ciprofloxacin and 100.00% were sensitive to erythromycin. Conclusions:The global sequence clusters of Cft isolates showed a great genetic diversity. Most of the people with Cft infection had basic immune diseases and might have eaten or had contact with reptiles. Notably, the Chinese domestic infection of ST46 and the emerging ST74 should arouse our more attention.
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Objective@#Establishing the mass spectrum library of a new Campylobacter- " C.fetus subsp.testudinum" for rapid species identification in clinical microbiology laboratory.@*Methods@#Illumina second generation sequencing platform 2000/miSeq was used to carry out high flux genome sequencing for the strains which were collected to establish mass spectrum library.The analysis oforthologous average nucleotide identity (OrthoANI) between collected strains and reference strains was performed at JAVA 8 operation environment. Then, the mass spectrums ofcollected strains andreference strains were acquired using MALDI-TOF MS. And the mass spectrum library of C. fetus subsp.testudinum. were established and verified.@*Results@#The OrthoANI analysis showed that the OrthoANI value of the collected strains and the reference strain C. fetus subsp.testudinum03-427 was 99.30%-99.96%, while the OrthoANI values of collected strains and C. fetus subsp.venerealisNCTC10354 orC.fetus subsp.fetus82-40 were 91.05%-92.26%. With reference to OrthoANI ≥ 95% as the basis for the determination of the same strain, the strains which collected to establish mass spectrum library was finally identified as " C. fetus subsp.testudinum" . The identification accuracy rate of the mass spectrum library was 100% (consistent with gene sequencing), and the confidence interval was 82.3%-99.9%, identification of the same strain is 100% reproducible.@*Conclusions@#The new" gold standard" based on high throughput sequencing and total genome analysis has provided the ideal reference value for the establishment of mass spectrum library.And the accurate and objective reference spectrum of the" C.fetus subsp.testudinum" provides a new platform for the rapid diagnosis of fetal Campylobacter infection. (Chin J Lab Med, 2018, 41: 583-588)
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Objective Reference standard of the RPOB(rifampin resistance)gene recommended by CLSI-MM18A(Interpretive Criteria for Identification of Bacteria and Fungi by DNA Target Sequencing) was used to evaluate the ability of MALDI-TOFMS techniques for the identification and classification of non-tuberculous Mycobacterium.Methods Fifty five clinicalstrains were collected from 2012 to 2016 with different sources.The RPOB gene was sequenced, and results were applied to phylogenetics analysis. MALDI-TOF MS technology was implemented to identify the strains, and cluster analysis was conducted based on protein fingerprint.The consistency of two methods for NTM identification and typing was evaluated.Results The RPOB gene method showed a good ability of identification(similarity>99.0%) and subtyping(to subspeciesof the complex level).The French BioMérieux MALDI-TOF MS identified 89.1% of 55 strains to genus level and 78.2% to species level.The phylogeneticsanalysis of protein fingerprint by SARAMS Premium software also showed good typing ability.Conclusions MALDI-TOF MS technology can identify and classify non-tuberculous Mycobacterium effectively,which is rapid and easy.It is complementary to RPOB gene method in laboratory application.
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Objective:To observe the effect of warm needling plus oral administration of rosuvastatin calcium tablets on blood lipids in cerebral infarction patients. Methods:A total of 125 eligible cases were randomly allocated into group A (n=42), group B (n=40) and group C (n=43). Cases in group A received warm needling plus oral administration of rosuvastatin calcium tablets, cases in group B received warm needling, whereas cases in group C received oral administration of rosuvastatin calcium tablets. Results:After treatment, the total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL-C) all dropped obviously in the three groups, with significant differences (allP0.05). After treatment, the changes of TC, TG and LDL-C in group A were significantly different from those in group B and group C (allP0.05). There were no between-group differences in HDL-C among the three groups (allP>0.05); the modified BI scores in groupA and groupB were significantly higher than that in group C (bothP0.05).After treatment, the total effective rate was significantly higher in group A than that in group B and group C (bothP0.05). Conclusion:Warm needling and oral administration of rosuvastatin calcium tablets both can adjust blood lipids effectively in cerebral infarction patients with a similar therapeutic efficacy, while the effect gets better based upon combining both methods; acupuncture-moxibustion plays an important role in the recovery of nerve functions in cerebral infarction patients.
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Objective To observe the therapeutic efficacy of low-frequency electrical stimulation (LFES) plus acupuncture in treating deglutition disorders after cerebral stroke. Method Seventy-six patients were randomized into two groups. The treatment group (38 cases) was intervened by LFES plus acupuncture;the control group (38 cases) was by LFES. Result The water drinking test scores were significantly changed after the treatment in both groups (P<0.05); after the intervention, there were significant differences in comparing the water drinking test score, improvement score, and therapeutic efficacy grading between the two groups (P<0.05). Conclusion LFES and acupuncture are both effective in treating deglutition disorders after cerebral stroke, and the combination of the two methods can enhance the clinical efficacy.
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Objective To evaluate the analysis capability of urine protein qualitative test between AX-4030 and Cobas U411 u-rine dry chemistry analyzer,and study on evaluating the performance of qualitative test.Methods According to Clinical and Laboratory Standards Institute(CLSI)EP12-A2 document,analyzed the bias and imprecision of urineprotein qualitative test between the Aution MAX AX-4030 and Roche CobasU411 system.Their C50 ,C5 ~C95 intervals and imprecision curves were compared.The protein of 310 specimens were simultaneously determined by both Cobas U411 and AX-4030,in order to eval-uate their concordance.Results C50 for AX-4030 system was less than that for Cobas U411;C5 ~C95 interval of AX-4030 system was narrower than CobasU411.The imprecision curve of AX-4030 system was steeper than Cobas U411.The com-parison of the two analysis systems showed that the concordance was 96.8%,the positive concordance was 82.7%,and the negative concordance was 99.6%.The 95% credibility interval (CI)was 94.2%~98.16% and the Kappa value was 0.88. Conclusion For the sensitivity and imprecision of urine protein test in the C50 critical value,the AX-4030 system was better than Cobas U411.The concordance of them in determining clinical specimens was pole-strength.The evaluation recommen-ded by the EP12-A2 document is practical and effective.
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Objective To evaluate the of feasibility of Roche e601 for detecting hepatitis B virus surface antigen(HBsAg).Meth-ods To evaluate the feasibility of Roche e601 for detecting hepatitis B virus surface antigen(HBsAg).Results The coefficient of variation(CV)of inter-run and between-run from low and high value specimens was lower than the requirements of manufacturers;the negative and positive coincidence rates in detecting 60 external quality assessment controls were 100%(40/40)and 100%(20 /20),respectively;the detection threshold values of the system was 0.01 IU/mL;the results detected by this detecting system in 40 samples,in which the concentration of HBsAg was 0.8