Phylogeny and virulence gene profile of Francisella salimarina / 中华微生物学和免疫学杂志

Objective:To study the molecular phylogeny and virulence gene profile of Francisella salimarina. Methods:Phylogenetic analysis of Francisella salimarina was performed based on the global genome data of related Francisella species on GenBank database. The consistency in phylogenetic analysis based on single marker genes (such as 16S rRNA gene, rpoB gene and mdh gene) and the core genome as compared. Virulence genes and antibiotic resistance genes were annotated using the virulence factor database (VFDB) and the Comprehensive Antibiotic Resistance Database (CARD), respectively. The virulence of Francisella salimarina was analyzed with a Galleria mellonella (greater wax moth) infection model using Francisella philomiragia ATCC 25015 T as reference strain. Results:The phylogenetic analysis revealed that Francisella salimarina was closely related to Francisella philomiragia. The phylogenetic tree based on mdh gene was highly similar to that based on the core genome. Francisella salimarina could be differentiated from other related species by 16S rRNA gene or mdh gene, with the latter being more accurate. Eight Francisella salimarina strains carried multiple virulence genes, mainly involved in secretion, adhesion, immune regulation, motility and stress survival. Moreover, beta-lactam resistance gene blaFPH was identified in all eight strains. Francisella salimarina showed high lethality in the Galleria mellonella infection model, which was similar to Francisella philomiragia ATCC 25015 T. Conclusions:Francisella salimarina was a rare pathogen with similar pathogenicity to Francisella philomiragia. The mdh gene could be used as a molecular target for rapid identification of Francisella salimarina.

Identification and phylogenetic analysis of two clinical isolates of Chryseobacterium / 中华微生物学和免疫学杂志

Objective:To analyze the biological characteristics, phylogenic features and clinical significance of SQ219 and SQ220 isolated from clinical sputum and midstream urine specimens.Methods:The culture and biochemical characteristics of the two strains were observed. VITEK2 System, drug sensitivity testing and MALDI-TOF mass spectrometry were used for bacterial identification. Phylogenetic analysis based on 16S rRNA and core genome was performed. The average nucleotide identity (ANI) based on whole genome sequences was calculated.Results:SQ219 and SQ220 were Gram-stain-negative, aerobic, catalase- and oxidase-positive, and non-motile bacteria. Their optimum growth was observed in NaCl-free medium at 30℃ and pH7. Flexirubin-type pigments were produced by SQ220 on Colombia blood agar, but not by SQ219. Both SQ219 and SQ220 were resistant to aztreonam, amikacin, tobramycin and colistin, which was consistent with the drug resistance phenotype of genus Chryseobacterium. The genome sequences of SQ219 and SQ220 were 5.08 Mb and 4.80 Mb in length, and the G+ C contents were 36.72% and 36.36%, respectively. Both strains carried β-lactam resistance gene ( blaCGA). 16S rRNA phylogenetic analysis showed that SQ219 and SQ220 were closely related to Chryseobacterium gambrini DSM18014 T with the similarities of 98.93% and 98.36%, respectively. Core genome phylogenetic analysis revealed that SQ219 and SQ220 were highly homologous to Chryseobacterium gambrini DSM18014 T. However, the ANI values between the two strains and Chryseobacterium gambrini DSM18014 T were 92.49% and 93.27%, respectively, below the threshold for prokaryotic species identification. Conclusions:Based on the phenotypic and phylogenetic data, SQ219 and SQ220 represent a novel species of the genus Chryseobacterium. This study would help promote the understanding of the evolution of Chrysobacterium and provide reference for the identification of new species of Chrysobacterium.

Structural control and characterization of hierarchically structured fibrous scaffolds / 中国修复重建外科杂志

Objective: To prepare hierarchically structured fibrous scaffolds with different morphologies, and to explore the additional dimensionality for tuning the physicochemical properties of the scaffolds and the effect of their hemocompatibility and cytocompatibility. Methods: Electrospinning poly (e-caprolactone) (PCL)/polyvinylpyrrolidone (PVP) bicomponent fibers (PCL∶PVP mass ratios were 8∶2 and 5∶5 respectively), and the surface porous fibrous scaffolds were prepared by extracting PVP components. The scaffolds were labeled PCL-P8 and PCL-P5 respectively according to the mass ratio of polymer. In addition, shish-kebab (SK) structured scaffolds with different kebab sizes were created by solution incubation method, which use electrospun PCL fibers as shish while PCL chains in solution crystallizes on the fiber surface. The PCL fibrous scaffolds with smooth surface was established as control group. The hierarchically structured fibrous scaffolds were characterized by field emission scanning electron microspore, water contact angle tests, and differential scanning calorimeter (DSC) experiments. The venous blood of New Zealand white rabbits was taken and hemolysis and coagulation tests were used to characterize the blood compatibility of the scaffolds. The proliferation of the pig iliac artery endothelial cell (PIEC) on the scaffolds was detected by cell counting kit 8 (CCK-8) method, and the biocompatibility of the scaffolds was evaluated. Results: Field emission scanning electron microscopy showed that porous morphology appeared on the surface of PCL/PVP bicomponent fibers after extracting PVP. In addition, SK structure with periodic arrangement was successfully prepared by solution induction, and the longer the crystallization time, the larger the lamellar size and periodic distance. The contact angle and DSC measurements showed that when compared with smooth PCL fiber scaffolds, the crystallinity of PCL surface porous fibrous scaffolds and PCL-SK fibrous scaffolds increased, while the hydrophobicity of PCL-SK fibrous scaffolds increased, but the hydrophobicity of PCL porous scaffolds did not change significantly. The hemolysis test showed that the hemolysis rate of PCL surface porous fibrous scaffolds and PCL-SK fibrous scaffolds was higher than that of PCL fibrous scaffolds. According to American Society of Materials and Tests (ASTM) F756-08 standard, all scaffolds were non-hemolytic materials and were suitable for blood contact materials. Coagulation test showed that the coagulation index of PCL surface porous fibrous scaffolds and PCL-SK fibrous scaffolds was higher than that of PCL fibrous scaffolds at 5 and 10 minutes of culture. CCK-8 assay showed that both hierarchically structured fibrous scaffolds were more conducive to PIEC proliferation than PCL fibrous scaffold. Conclusion: Based on electrospinning technology, solution-induced and blend phase separation methods can be used to construct multi-scale fiber scaffolds with different morphologies, which can not only regulate the surface physicochemical properties of the scaffolds, but also have good blood compatibility and biocompatibility. The hierarchically structured fibrous scaffolds have high application potential in the field of tissue engineering.

Application of OrthoANI combined with MALDI-TOF MS in identification and typing of Campylobacter fetus / 中华检验医学杂志

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)

Identification and classification of non-tuberculosis Mycobacterium by matrix-assisted laser desorption ionization-time of flight mass spectrometry / 中华检验医学杂志

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.

Empathy of obstetric nurses and the related factors / 现代临床护理

Objective To investigate the empathy of obstetric nurses and the related factors.Methods Chinese Version of the Jefferson Scale of Empathy was used to investigate the empathy of 80 obstetric nurses in our hospital and the related factors.Results The score on empathy of obstetric nurses was 65~137,averaged 108.87±10.62 and ranked at the medium level.The empathy was mainly influenced by whether the nurses had studied the empathy-related courses and the satisfaction with their occupation and working environment.Conclusion The empathy of obstetric nurses is at the medium level.Their empathy is mainly influenced by whether the nurses have studied the empathy-related courses and the satisfaction of their occupation and working environment.In addition to promote the training,the nursing manager should improve the satisfaction of the occupation and working environment.

A comparative study on Nevin and AJCC staging system for gallbladder carcinoma / 中华普通外科杂志

Objective To evaluate the clinical value of Nevin and AJCC staging system for gallbladder carcinoma. Methods In this study 90 patients diagnosed as gallbladder carcinoma underwent operation in Renji Hospital from February 2000 to October 2006. Patients were staged according to Nevin and AJCC staging system. The difference of survival rate, tumor resection rate, ratio of tumor-free resection margin and surgical procedures were analyzed. Results The survival rate, tumor resection rate and ratio of tumor-free resection margin decreased progressively with increasing Nevin and AJCC stage (P < 0. 05). There was no significant difference between the survival rate for Nevin Ⅲ or Ⅳ patients undergoing radical resection and simple cholecystectomy (P > 0. 05). In Nevin Ⅴ patients, the survival rate for radical and extensive radical resection patients was higher than for palliative patients (P < 0. 05). In AJCC Ⅲ patients, the survival rate for radical patients was significant higher than for palliative patients (P < 0. 05). Nosignificant difference was found between radical and palliative patients in survival time in AJCC Ⅳ (P > 0. 05). 52 patients in AJCC Ⅲ and Ⅳ were staged to Nevin Ⅴ according to Nevin staging system. The survival rate and resectable rate for the patients in AJCC Ⅲ were higher than in AJCC Ⅳ (P = 0. 0001, 0. 001 respectively). The rate of radical operation in AJCC Ⅲ was higher (P = 0. 001), and the rate of palliative operation in AJCC Ⅳ was higher (P = 0. 001). Conclusion Both Nevin and AJCC staging system are useful in the judgement of survival, reeectability, ratio of tumor-free resection margin and the optimal operation. AJCC staging system is more applicable for gallbladder carcinoma patients at advanced stage in terms of predicting prognosis.

Transmission disequilibrium between congenital dislocation of the hip and homeobox-containing genes* / 中国组织工程研究

BACKGROUND: Research shows that genetic factors are an important component of the congenital dislocation of the hip(CDH) . However, no susceptibility genes have been identified by now. The homebox-containing (HOX) genes that regulate the embryogenesis and vertebrate limb development may play a role in the pathogenesis of CDH.OBJECTIVE: To investigate whether a correlation exists between CDH and the Hox genes.DESIGN: Controlled study associated with family.SETTING: Department of aevelopmental pediatrics, genetic laboratory, department of pediatric orthopaedics in an affiliated hospital of a university.PARTICIPANTS: All the 101 CDH patients and their parents (altogether 303 members) were the in-patients from the Department of Pediatric Orthopaedics of the Second Clinical College of China Medical University; from December 1999 to January 2001. All the patients presented typical clinical manifestations and were diagnosed by X rays and operations for confirmation.METHODS: Four microsatellite markers D7S1808, D17S1820, D12S1686 and Hox4EP were selected in the chromosome regions of7p14 - 15, 17q21, 12q13and 2q31 where Hox A, Hox B, Hox C and Hox D genes which regulate the embryonic limb development reside respectively. Genotypes of 303 members in 101CDH families were analyzed by the techniques of polymerase chain reaction (PCR) and denaturing polyacrylamide gel electrophoresis. Then transmission disequilibrium test(TDT) was performed to analyze the data of genotypes.MAIN OUTCOME MEASURES: The genotypes of four microsatellite markers D7S1808, D17S1820, D12S1686 and Hox4EP in every CDH family including one child and parents; transmission disequilibrium test between transmission alleles and non-transmission alleles.RESULTS: Transmission disequilibrium was found between CDH and allele 7 of D7S1808(X2 = 6. 045, P = 0. 014) among a total of 10 alleles detected, between CDH and allele 4 of D17S1820(X2 =6. 025, P =0. 014) among a total of 12 alleles detected, between CDH and allele 4 of Hox4Ep (X2 = 6. 461, P =0.011) among a total of 16 alleles detected. But no transmission disequilibrium was found between CDH and D12S1686(X2 = 6. 171,P =0. 965) with 16 alleles detected.CONCLUSION: CDH may be related to Hox A, Hox B, Hox D genes, and Hox A, Hox B, Hox D genes may be susceptibility genes in CDH.

Expression of midkine in benign, premalignant and malignant vulvar tumors / 中国医学科学杂志(英文版)

<p><b>OBJECTIVE</b>To clarify the role of midkine (MK) in vulvar carcinogenesis though examination of its expression in vulvar lesions including vulvar condyloma acuminata (VCA), vulvar intraepithelial neoplasia (VIN) and vulvar squamous cell carcinomas (VSCC), and to analyze the relationship between MK expression and human papilloma virus (HPV) infection.</p><p><b>METHODS</b>Thirty VSCC, 15 VIN and 10 VCA patients were studied by streptavidin-biotin-immunoperoxidase method. MK expression was compared with clinicopathologic features of vulvar tumors.</p><p><b>RESULTS</b>MK was expressed in 26 of 30 VSCC (87%), 3 of 5 VIN III and all VCA samples, whereas no MK expression was detected in the VIN I-II samples or in normal epithelium. The difference of MK expression between VIN III and VSCC was statistically significant (P < 0.05). MK was more intensely expressed in differentiated-type (well differentiated and moderately differentiated) VSCC than in undifferentiated-type (poorly differentiated) VSCC. There was no statistically significant correlation between MK expression and clinical stage, lymph node metastasis and HPV infection in VSCC. MK expression were observed in all HPV-positive specimens including 2 VSCC, 1 VIN III and all VCA.</p><p><b>CONCLUSIONS</b>MK gene expression may be a late event in vulvar squamous cell malignant transformation, and may be associated with vulvar tumor cell differentiation. HPV-positive vulvar tumors expressed MK protein.</p>