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Objective@#To investigate the antimicrobial resistance and molecular epidemiology of foodborne Yersinia (Y.) enterocolitica in Pudong New District of Shanghai.@*Methods@#Four kinds of raw food samples were collected in retail circulation sites in Pudong from 2012 to 2016. Cold enrichment method was used to isolate Y. enterocolitica and further detection of biotype, serotype, virulent genes, antimicrobial susceptibility of the isolates and pulsed field gel electrophoresis (PFGE) were conducted.@*Results@#A total of 3 900 raw food samples were collected during this period, including poultry product (n=590), livestock product (n=1 074), aquatic product (n=1 488), vegetable (n=748), in which 111 (2.8%) were contaminated by Y. enterocolitica. The detection rates of Y. enterocolitica in poultry product samples (5.3%, 31/590) and livestock product samples (4.5%, 48/1 074) were higher than those in aquatic product samples (1.6%, 24/1 488) and vegetable samples (1.1%, 8/748). The predominant biotype was 1A (95.5%) and predominant serotype was O∶8 (42.3%). All the strains lacked ail, ystA, yadA and virF genes, which encoded pathogenic Y. enterocolitica. Seventy six (68.5%) strains harbored ystB gene, in which 35 (31.5%) belonged to 1A/O∶8/ystB pattern. Most strains were resistant to ampicillin (74.8%) and amoxicillin/clavulanic acid (70.3%), and non-sensitive rate to Cefoxitin was over 50.0%. No third generation cephalosporin or fluoroquinolone resistant strains were detected, but 38.7% (43/111) strains were multidrug resistant (MDR). Serotype O∶8 and O∶5 strains had 44 and 18 PFGE patterns, respectively.@*Conclusions@#The main foodborne exposure sources of Y. enterocolitica in raw food were poultry and livestock products in Pudong New District. 1A/O∶8/ystB was the predominant pattern with potential pathogenicity despite lacks of typical pathogenic virulent genes. The antimicrobial resistant rates of Y. enterocolitica were at a low level, but MDR strains still existed. Molecular types of the isolates showed highly genetic diversity.
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Objective@#To understand the in vitro growth characteristics of Cryptococcus gattii VGⅠ and VGⅡ isolated in China and the diversity in their virulence to Galleria mellonella.@*Methods@#Based on the results of multilocus sequence typing for eight strains of Cryptococcus gattii isolated in China, the strains were cultured in vitro to draw growth curves, observe the melanin production and measure the capsule thickness. The median lethal time (LT50) and median lethal dose (LC50) at 48 h of Cryptococcus gattii were calculated using Galleria mellonella infection test. Fourteen strains of Cryptococcus neoformans were studied for comparison.@*Results@#The eight Cryptococcus gattii strains were six VGⅠ and two VGⅡ. The growth curves of Cryptococcus gattii VGⅠ and VGⅡ were similar to that of Cryptococcus neoformans when culture at 30℃. The total number for each of them could reach 108 CFU/ml at 96 h under 30℃. However, the total number at any time point at 37℃ was less than that at 30℃. There was no significant difference in the amount of melanin produced by Cryptococcus neoformans under 30℃ and 37℃, but both VGⅠand VGⅡ types of Cryptococcus gattii could produce more amount of melanin under 37℃ than under 30℃. The ratio of capsule/cell wall diameter of Cryptococcus gattii VGⅠwas greater at 37℃ than that at 30℃ with statistical significance (P<0.001). Cryptococcus neoformans showed the longest LT50, followed by VGⅠand VGⅡ types of Cryptococcus gattii. The LT50 of Cryptococcus gattii VGⅡ at the concentration of 1×106 CFU/ml was 72 h, and its LC50 at 48 h was 1×108 CFU/ml.@*Conclusions@#Like Cryptococcus neoformans, Cryptococcus gattii VGⅠ and VGⅡ grew faster under 30℃ than under 37℃, but more melanin was produced and thicker capsule was formed under 37℃ than under 30℃. Among Cryptococcus neoformans and VGⅠ and VGⅡ types of Cryptococcus gattii, Cryptococcus gattii VGⅡ showed the shortest LT50 and the strongest virulence to Galleria mellonella.
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Objective To understand the in vitro growth characteristics of Cryptococcus gattii VGⅠ and VGⅡ isolated in China and the diversity in their virulence to Galleria mellonella. Methods Based on the results of multilocus sequence typing for eight strains of Cryptococcus gattii isolated in China, the strains were cultured in vitro to draw growth curves, observe the melanin production and measure the capsule thickness. The median lethal time (LT50 ) and median lethal dose (LC50 ) at 48 h of Cryptococcus gattii were calculated using Galleria mellonella infection test. Fourteen strains of Cryptococcus neoformans were studied for comparison. Results The eight Cryptococcus gattii strains were six VGⅠ and two VGⅡ. The growth curves of Cryptococcus gattii VGⅠ and VGⅡ were similar to that of Cryptococcus neoformans when culture at 30℃ . The total number for each of them could reach 108 CFU/ ml at 96 h under 30℃ . However, the total number at any time point at 37℃ was less than that at 30℃ . There was no significant difference in the amount of melanin produced by Cryptococcus neoformans under 30℃ and 37℃ , but both VGⅠand VGⅡtypes of Cryptococcus gattii could produce more amount of melanin under 37℃ than under 30℃ . The ratio of capsule/ cell wall diameter of Cryptococcus gattii VGⅠwas greater at 37℃ than that at 30℃ with statistical significance (P<0. 001). Cryptococcus neoformans showed the longest LT50 , followed by VGⅠand VGⅡtypes of Cryptococcus gattii. The LT50 of Cryptococcus gattii VGⅡ at the concentration of 1×106 CFU/ ml was 72 h, and its LC50 at 48 h was 1×108 CFU/ ml. Conclusions Like Cryptococcus neoformans, Cryptococcus gattii VGⅠ and VGⅡ grew faster under 30℃ than under 37℃ , but more melanin was produced and thicker capsule was formed under 37℃ than under 30℃ . Among Cryptococcus neoformans and VGⅠ and VGⅡtypes of Cryptococcus gattii, Cryptococcus gattii VGⅡ showed the shortest LT50 and the strongest virulence to Galleria mellonella.
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OBJECTIVE:To investigate the effects of neoadjunctive chemotherapy(NAC)of docetaxel and epirubicin com-bined with cyclophosphamide on clinical efficacy and tumor markers of breast cancer patients with different molecular types. METH-ODS:A total of 88 female patients with locally advanced breast cancer collected from our hospital during Jan. 2014-Jan. 2016 were divided into Luminal A type(23 cases),Luminal B type(21 cases),basal-like type(11 cases),HER2-over expressing type(18 cases)and normal breast-like type(15 cases)according to molecular type. All patients were given Docetaxel injection+Epirubicin hydrochloride injection+Cyclophosphamide for injection for consecutive 6 cycles(21 d as a cycle). Total response rates and patho-logical complete remission(pCR)rates were compared among breast cancer patients with different molecular types. The expression of serum tumor markers [CEA,CA125,CA153] were compared before and after treatment,and the occurrence of ADR was record-ed. RESULTS:Total response rate of 88 patients was 63.64%,among which that of basal-like breast cancer patients was 72.73%, significantly higher than other molecular types,with statistical significance(P0.05). The pCR rate of 88 patients was 27.27%,and that of basal-like breast cancer patients was the highest(45.45%). There was statistical significance in pCR rates of pairwise molecular type compari-son(P0.05). Before treatment,there was no statistical significance in the expression of CEA,CA125 and CA153 in breast can-cer patients with different molecular types(P>0.05). After treatment,the expression of CEA,CA125 and CA153 in different mo-lecular types were decreased significantly,with statistical significance(P0.05). There was no statistical significance in the incidence of ADR among different molecular types(P>0.05). CONCLUSIONS:NAC plan of docetaxel and epirubicin combined with cyclophospha-mide can reduce the expression of tumor markers and shows certain therapeutic efficacy for breast cancer patients with different mo-lecular types. Total response rate and pCR rate of basal-like type are better than those of other molecular types,so NAC plan is the preferred treatment for basal-like type breast cancer.
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[ Objective] To investigate the status quo of Campylobacter spp.infection in Shanghai and study its molecular characteristics and virulence and toxin genes. [ Methods ] Stool samples collected from diarrheal patients were cultured for bacterial pathogens using membrane filter method.The strains were identified by biochemical tests and PCR.PCR was applied to detect six virulence and toxin genes including flaA,cdtA,cdtB,cdtC,virB11,cadF.Pulsed-field gel electrophoresis ( PFGE) was carried out for subtyping. [Results] A total of 43 Campylobacter spp.(1.9%) were collected from 2 235 stool samples in Shanghai in 2014 including 41 Campylobacter jejuni isolates(95.3%) and 2 Campylobacter coli isolates(4.7%) .The data showed 100.0%(43/43) of the isolates were positive for flaA and cadF, and 93.0%(40/43) of the isolates positive for cdtA and cdtB.And 88.4%(38/43) of the isolates were posi-tive for cdtC.Only 7.0%(3/43) of the isolates were positive for virB11.Using PFGE, 43 Campylobacter jejuni and Campylobacter coli strains were subtyped into 6 clusters. [ Conclusion] The genes of flaA and cadF are ubiquitous on Campylobacter spp.isolates.The distribution of cdt gene cluster in Campylobacter spp.is high, while that of virB11 is low.The PFGE types of Campylobacter spp.isolated in Shanghai are of diversified and complicated features, which causes mainly sporatic diarrhea.
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PURPOSE: Human epidermal growth factor receptor 2 (HER2)-positive luminal B type comprises estrogen receptor (ER)-positive and HER2-positive cancers, and HER2-negative luminal B type comprises ER-positive cancers showing a Ki-67 labeling index > or =14% or progesterone receptor (PR) expression of <20% according to the St. Gallen consensus 2013. The current study aimed to classify intrinsic subtypes according to the St. Gallen consensus 2013 and determine the differences in clinicopathological parameters and survival outcomes among the molecular types, especially among the luminal types. METHODS: Assessment of molecular types was performed for 267 invasive ductal carcinomas. The differences in clinicopathological parameters, disease-free survival (DFS), and overall survival (OS) among the molecular types were analyzed. RESULTS: The luminal B type was the most prevalent, at 44.9%, followed by the luminal A, triple-negative (including basal-like type), and HER2 type, at 21.7%, 18.7%, and 14.6%, respectively. There were statistically significant differences in size (p=0.003), nodal status (p=0.046), histologic grade (p<0.001), p53 (p<0.001) and cyclooxygenase 2 (COX-2) positivity (p=0.002), recurrence (p=0.001) and death rates (p=0.036), DFS (p=0.002), and OS (p=0.039) among the molecular types. Significant differences in size (p=0.009), nodal metastasis (p=0.019), histologic grade (p<0.001), p53 positivity (p=0.001), and PR expression (p<0.001) were noted between the luminal A and B types. Among the luminal B type cancers, the distributions of ER and PR scores showed significant differences (p=0.003, p=0.003). p53 positivity in the luminal B type cancers was related to shortened DFS (p=0.034). In luminal type cancers, COX-2 positivity was related to longer DFS (p=0.026). CONCLUSION: Different management guidelines should be considered for the luminal A and luminal B breast cancer types. Positive p53 expression in luminal B type cancers and negative COX-2 expression in luminal type cancers seem to be related to poor clinical outcome.
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Humans , Breast Neoplasms , Breast , Carcinoma, Ductal , Consensus , Cyclooxygenase 2 , Disease-Free Survival , Estrogens , Ki-67 Antigen , Mortality , Neoplasm Metastasis , Phenobarbital , ErbB Receptors , Receptors, Progesterone , RecurrenceABSTRACT
A criptococose é causada por duas espécies distintas de leveduras capsuladas da divisão dos basidiomicetos, Cryptococcus neoformans e Cryptococccus gattii, as quais causam infecções invasivas emergentes de significativa letalidade, seja em imunodeprimidos ou imunocompetentes. No Brasil, a criptococose por C. gattii tem caráter endêmico, ocorrendo principalmente em indivíduos HIV negativos. O caráter epidêmico deste agente tem sido evidenciado por surtos em animais, sendo o mais significativo a epidemia ocorrida em Vancouver, Canadá, atingindo humanos e animais desde 1999...
Cryptococcosis is caused by two distinct species of capsulated basidiomycetes yeasts, Cryptococcus neoformans and Cryptococccus gattii, which are responsible for invasive infections with significant lethality, either in immunocompetent or immunocompromised individuals. In Brazil, cryptococcosis by C. gattii is endemic, occurring mainly in HIV negative individuals. The epidemic nature of this agent has been reported by outbreaks in animals and human in Vancouver, Canada, which is the most significant outbreak and started in 1999...
Subject(s)
Animals , AIDS-Related Opportunistic Infections , Cryptococcus gattii , Cryptococcosis/classification , Cryptococcosis/epidemiology , Genotype , Multilocus Sequence Typing , Polymerase Chain Reaction , Polymorphism, Restriction Fragment LengthABSTRACT
Cryptococcal infection is primarily caused by two species, Cryptococcus neoformans and C. gattii. Between the two species, C. neoformans var. grubii is the major causative agent of cryptococcosis in Asia. We investigated the molecular characteristics of 46 isolates of C. neoformans from patients with cryptococcosis between 2008 and 2012 in Seoul, Korea. All the isolates were determined to be C. neoformans var. grubii (serotype A), mating type MATalpha, and molecular type VNI by PCR-restriction fragment length polymorphism of the URA5 gene. Multilocus sequencing type (MLST) analysis using the International Society of Human and Animal Mycoses (ISHAM) consensus MLST scheme identified two sequence types (ST). Out of the 46 strains, 44 (95.7%) were identified as ST5, and remaining 2 were identified as ST31. Our study revealed that the clinical strains of C. neoformans in Korea are genetically homogeneous with the VNI/ST5 genotypes, and new appearance of VNI/ST31 genotype may serve as an important indicator of global genetic analysis.
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Animals , Humans , Asia , Consensus , Cryptococcosis , Cryptococcus neoformans , Cryptococcus , Genotype , Korea , Molecular Epidemiology , Mycoses , SeoulABSTRACT
Aim: Considering the geographic expansion of Cryptococcus gattii, the aim of this study was to investigate hollows of living trees as a reservoir of C. gattii in Rio de Janeiro, Brazil. Place and Duration of the Study: In an urban quarter of Rio de Janeiro city, 80 samples of decaying wood were collected. In addition, 85 decaying wood samples were collected in the wild rainforest. The samples were analyzed at the Mycology Laboratory, Evandro Chagas Clinical Research Institute, Oswaldo Cruz Foundation, from 2008-2010. Methodology: Samples were collected by scraping the inner decaying wood of the hollows of the trunks of each tree. Pathogenic Cryptococcus species were identified by: brown colonies on niger seed agar (NSA) medium, thermotolerance at 35ºC, cycloheximide sensitivity, carbon and nitrogen assimilation tests performed by 32-Vitek System (Vitek ICB, bioMeriux, Durham, EUA). Canavanine-glycine-bromothymol blue medium (CGB) was used to determine the species of the isolates and the genotypes were determined by restriction fragment length polymorphism of URA5 gene. Results: After plating the samples on NSA, 584 colonies were obtained from the urban quarter. C. gattii VGI was identified in 98% of colonies, followed by C. neoformans VNI 2%. The positivity of the urban area was 7.8%. The concentrations of the fungi in hollows of ficus trees ranged from 50 to 56,250 colony-forming units per gram of sample (CFU/g). Conclusions: For the first time in Rio de Janeiro C. gattii VGI was isolated in a hollow of living tree.
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Cyptococcosis is generally caused by Cryptococcus neoformans, the opportunistic agent which has two species such as C. neoformans and C. gattii. Both C. neoformans and C. gattii species contain a number of genetically diverse subgroups that can be differentiated by various molecular typing methods. We conducted a molecular epidemiological analysis of 30 clinical isolates of the C. neoformans from cryptococcosis patients who had been hospitalized between 2008 and 2010 in medical centers located in Seoul and Busan in Korea. To determine the genetic diversity, 30 strains of C. neoformans were typed using PCR fingerprinting with the microsatellite specific primer of the phage M13 and the restriction fragment length polymorphism (RFLP) of orotidine monophosphosphate pyrophosphorylase (URA5) gene. All isolates were identified as serotype A, mating type MATa and molecular type VNI. The random amplified polymorphic DNA (RAPD) profiles obtained by using two primers revealed a single pattern. Our study shows that 30 strains of clinical C. neoformans are genetically homogeneous, with all of the isolates were molecular type VN1, serotype A, mating type MATa.
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Humans , Bacteriophage M13 , Cryptococcosis , Cryptococcus , Cryptococcus neoformans , Dermatoglyphics , DNA , Genetic Variation , Korea , Microsatellite Repeats , Molecular Typing , Polymerase Chain Reaction , Polymorphism, Restriction Fragment Length , UridineABSTRACT
Objective To describe the phenotype and molecular characteristics of Vibrio (V.) cholerae strains isolated in Shanghai,from 1962 to 2011.Methods K-B test was used to investigate the antibiotic resistance of V.cholerae strains.PCR was applied to detect seven virulence-related genes including cholera toxin (ctxA),zonula oecludens toxin (zot),accessory cholera enterotoxin (ace),hemolysin (hlyA),toxin-coregulated pilus (tcpA) outer membrane protein (ompU) and the regulatory protein genes (toxR).Genetic relation was assessed by pulsed-field gel electrophoresis (PFGE) and the patterns were clustered by BioNumerics software.Results V.cholerae strains isolated from 1962 to 1996 were sensitive to most of the antibiotics.However,the strains isolated from 2005 to 2011 were resistant to many antibiotics.V.cholerae O 139 group showed higher prevalence of resistance to several antibiotics compared with O l group,and the resistance rate of the O139 toxigenic isolates was higher than that of the non-toxigenic isolates.Most of the O1 strains isolated from 2005 to 2011 were non-toxigenic while O139 strains isolated from 2005 to 2011 were almost toxigenic.There were no strains ofctxA+ detected from the rivers from 2005 to 2011.Main gene type of the O1 strains detected from the aquatic products was hlyA+toxR+ompU+,while that of the O139 strains was hlyA+toxR+ompU+ ctxA + ace +zot + tcpA +.Using PFGE,222 V.cholerae strains were subtyped into 121 molecular types.O139 strains were divided to three clusters and O1 strains to five clusters.Conclusion The characteristics of V.cholerae strains isolated in Shanghai from 1962 to 2011 showed great changes,suggesting that more attention should be paid to the multiplication on antibiotic resistance of V.cholerae strains.
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Cryptococcus gattii causes life-threatening yeast infection in the pulmonary and central nervous systems of humans and animals, and traditionally has been considered to restrict into the tropical and subtropical areas. Despite rare incidence of cryptococcosis caused by C. gattii in Korea, three strains of C. gattii isolated from cryptococcosis patients between 1993 and 2010 were identified. To determine the genetic diversity, 3 strains of C. gattii were typed using PCR fingerprinting with primer M13 and the restriction fragment length polymorphism (RFLP) of orotidine monophosphosphate pyrophosphorylase (URA5) gene. All isolates were identified as serotype B and MATalpha mating type. The molecular types of each strain, on the other hand, turned out to be distinct belonging to VGI, VGII or III types, respectively. Although the travel histories of the patients were not available, clinical C. gattii strains isolated in Korea may represent the diverse molecular types existing worldwide.