RESUMEN
Patent ductus arteriosus (PDA) is a common congenital heart disease. CITED2 plays an important role in the development of the heart, and genetic variants in its coding region are significantly associated with cardiac malformations. However, the role of variants in the promoter region of CITED2 in the development of PDA remains unclear. We extracted the peripheral blood of 646 subjects (including 353 PDA patients and 293 unrelated healthy controls) for sequencing. We identified 13 promoter variants of the CITED2 gene (including 2 novel heterozygous variants). Of the 13 variants, 10 were found only in PDA patients. In mouse cardiomyocytes (HL-1) and rat cardiac myocytes (RCM), the transcriptional activity of the CITED2 gene promoter was significantly changed by the variants (p < 0.05). The results of the experiments of electrophoretic mobility indicated that these variants may affect the transcription of the CITED2 gene by influencing the binding ability of transcription factors. These results, combined with the JASPAR database analysis, showed that the destruction/production of transcription factor binding sites due to the variants in the promoter region of the CITED2 gene may directly or indirectly affect the binding ability of transcription factors. Our results suggest for the first time that variants at the CITED2 promoter region may cause low expression of CITED2 protein related to the formation of PDA.
Asunto(s)
Conducto Arterioso Permeable , Cardiopatías Congénitas , Humanos , Animales , Ratones , Ratas , Conducto Arterioso Permeable/genética , Conducto Arterioso Permeable/metabolismo , Cardiopatías Congénitas/genética , Factores de Transcripción/genética , Miocitos Cardíacos/metabolismo , Regiones Promotoras Genéticas , Proteínas Represoras/genética , Transactivadores/genética , Transactivadores/metabolismoRESUMEN
CONTEXT: Hyperoside (Hyp), one of the active flavones from Rhododendron (Ericaceae), has beneficial effects against cerebrovascular disease. However, the effect of Hyp on vasodilatation has not been elucidated. OBJECTIVE: To explore the effect of Hyp on vasodilatation in the cerebral basilar artery (CBA) of Sprague-Dawley (SD) rats suffering with ischaemic-reperfusion (IR) injury. MATERIALS AND METHODS: Sprague-Dawley rats were randomly divided into sham, model, Hyp, Hyp + channel blocker and channel blocker groups. Hyp (50 mg/kg, IC50 = 18.3 µg/mL) and channel blocker were administered via tail vein injection 30 min before ischaemic, followed by 20 min of ischaemic and 2 h of reperfusion. The vasodilation, hyperpolarization, ELISA assay, haematoxylin-eosin (HE), Nissl staining and channel-associated proteins and qPCR were analysed. Rat CBA smooth muscle cells were isolated to detect the Ca2+ concentration and endothelial cells were isolated to detect apoptosis rate. RESULTS: Hyp treatment significantly ameliorated the brain damage induced by IR and evoked endothelium-dependent vasodilation rate (47.93 ± 3.09% vs. 2.99 ± 1.53%) and hyperpolarization (-8.15 ± 1.87 mV vs. -0.55 ± 0.42 mV) by increasing the expression of IP3R, PKC, transient receptor potential vanilloid channel 4 (TRPV4), IKCa and SKCa in the CBA. Moreover, Hyp administration significantly reduced the concentration of Ca2+ (49.08 ± 7.74% vs. 83.52 ± 6.93%) and apoptosis rate (11.27 ± 1.89% vs. 23.44 ± 2.19%) in CBA. Furthermore, these beneficial effects of Hyp were blocked by channel blocker. DISCUSSION AND CONCLUSIONS: Although Hyp showed protective effect in ischaemic stroke, more clinical trial certification is needed due to the difference between animals and humans.
Asunto(s)
Antineoplásicos , Isquemia Encefálica , Daño por Reperfusión , Accidente Cerebrovascular , Humanos , Ratas , Animales , Ratas Sprague-Dawley , Canales Catiónicos TRPV/metabolismo , Células Endoteliales , Isquemia Encefálica/tratamiento farmacológico , Accidente Cerebrovascular/metabolismo , Vasodilatación , Antineoplásicos/farmacología , Daño por Reperfusión/tratamiento farmacológico , Daño por Reperfusión/prevención & control , Daño por Reperfusión/metabolismoRESUMEN
The aim of this study is to explore the mechanism of ligustilide, the main active constituent of essential oils of traditional Chinese medicine Angelicae Sinensis Radix, on alleviating oxygen-glucose deprivation/reperfusion(OGD/R) injury in PC12 cells from the perspective of ferroptosis. OGD/R was induced in vitro, and 12 h after ligustilide addition during reperfusion, cell viability was detected by cell counting kit-8(CCK-8) assay. DCFH-DA staining was used to detect the level of intracellular reactive oxygen species(ROS). Western blot was employed to detect the expression of ferroptosis-related proteins, glutathione peroxidase 4(GPX4), transferrin receptor 1(TFR1), and solute carrier family 7 member 11(SLC7A11), and ferritinophagy-related proteins, nuclear receptor coactivator 4(NCOA4), ferritin heavy chain 1(FTH1), and microtubule-associated protein 1 light chain 3(LC3). The fluorescence intensity of LC3 protein was analyzed by immunofluorescence staining. The content of glutathione(GSH), malondialdehyde(MDA), and Fe was detected by chemiluminescent immunoassay. The effect of ligustilide on ferroptosis was observed by overexpression of NCOA4 gene. The results showed that ligustilide increased the viability of PC12 cells damaged by OGD/R, inhibited the release of ROS, reduced the content of Fe and MDA and the expression of TFR1, NCOA4, and LC3, and improved the content of GSH and the expression of GPX4, SLC7A11, and FTH1 compared with OGD/R group. After overexpression of the key protein NCOA4 in ferritinophagy, the inhibitory effect of ligustilide on ferroptosis was partially reversed, indicating that ligustilide may alleviate OGD/R injury of PC12 cells by blocking ferritinophagy and then inhibiting ferroptosis. The mechanism by which ligustilide reduced OGD/R injury in PC12 cells is that it suppressed the ferroptosis involved in ferritinophagy.
Asunto(s)
Ferroptosis , Animales , Ratas , Células PC12 , Ferroptosis/genética , Especies Reactivas de Oxígeno , Factores de Transcripción , GlutatiónRESUMEN
BACKGROUND: Atrial septal defect (ASD) is a common type of congenital heart disease. A gene promoter plays pivotal role in the disease development. This study was designed to investigate the pathological role of variants of the ISL1 gene promoter region in ASD patients. METHODS: Total DNA extracted from 625 subjects, including 332 ASD patients and 293 healthy controls, was sequenced to identify variants in the promoter region of ISL1 gene. Further functional analyses of the variants were performed with dual luciferase reporter assay and electrophoretic mobility shift assay (EMSA). All possible binding sites of transcription factor affected by the identified variants were predicted using the JASPAR database. RESULTS: Four variants in the ISL1 gene promoter were found only in patients with ASD by sequencing. Three of the four variants [g.4923 G > C (rs541081886), g.5079 A > G (rs1371835943) and g.5309 G > A (rs116222082)] significantly decreased the transcriptional activities compared with the wild-type ISL1 gene promoter (p < 0.05). The EMSA revealed that these variants [g.4923 G > C (rs541081886), g.5079 A > G (rs1371835943) and g.5309 G > A (rs116222082)] in the ISL1 gene promoter affected the number and affinity of binding sites of transcription factors. Further analysis with the online JASPAR database demonstrated that a cluster of putative binding sites for transcription factors may be altered by these variants. CONCLUSIONS: These sequence variants identified from the promoter region of ISL1 gene in ASD patients are probably involved in the development of ASD by affecting the transcriptional activity and altering ISL1 levels. Therefore, these findings may provide new insights into the molecular etiology and potential therapeutic strategy of ASD.
Asunto(s)
Defectos del Tabique Interatrial , Humanos , Defectos del Tabique Interatrial/genética , Regiones Promotoras Genéticas , Factores de Transcripción/genéticaRESUMEN
This paper explored the protective effect of total flavonoids of Rhododendron simsii(TFR) on focal cerebral ischemia-reperfusion injury(CIRI) in rats and its relationship with the store-operated calcium entry(SOCE) pathway regulated by stromal intera-ction molecule(STIM) and calcium release-activated calcium modulator(Orai).Rats were randomly assigned into the sham group, model(middle cerebral artery occlusion, MCAO) group, TFR(60 mg·kg~(-1)) group, TFR(60 mg·kg~(-1))+SOCE pathway inhibitor 2-aminoethoxydiphenyl borate(2-APB, 2.5 mg·kg~(-1)) group, and 2-APB(2.5 mg·kg~(-1)) group.The rats in the sham group and MCAO group were administrated with normal saline, and those in the TFR group and TFR+2-APB group were administrated with TFR(60 mg·kg~(-1)) by gavage for 14 days until sampling.The rats in the 2-APB group and TFR+2-APB group were intraperitoneally injected with 2-APB(2.5 mg·kg~(-1)) after operation.The levels of interleukin-1(IL-1), interleukin-6(IL-6), and tumor necrosis factor-alpha(TNF-α) in serum were measured by ELISA.The cerebral infarction and the pathological status of ischemic brain tissue were detected via TTC staining and HE staining, respectively.The protein and mRNA levels of STIM1, STIM2, Orai1, cysteinyl aspartate specific proteinase 3(caspase-3), and protein kinase B(PKB) in brain tissue were respectively determined by Western blot and RT-qPCR.The growth of brain neurons in each group was observed via immunofluorescence method.The results showed that compared with the MCAO group, TFR lowered the levels of IL-1, IL-6 and TNF-α in serum and the score of neurological function, ameliorated the pathological injury of brain tissue, and decreased the infarct size.Moreover, TFR up-regulated the mRNA and protein levels of STIM1, STIM2, Orai1, and PKB, down-regulated those of caspase-3 in brain tissue, and increased the double-labeled positive cells under fluorescence microscope.However, the above effects were significantly weakened by the addition of 2-APB, a SOCE inhibitor.The results suggested that TFR may play a protective role against focal cerebral ischemia-reperfusion injury by up-regulating the expression of SOCE-related signal molecules, promoting neurogenesis around the ischemic area, improving the survival state of neurons, and redu-cing the activity of inflammatory mediators.
Asunto(s)
Isquemia Encefálica , Daño por Reperfusión , Rhododendron , Animales , Isquemia Encefálica/tratamiento farmacológico , Isquemia Encefálica/genética , Isquemia Encefálica/metabolismo , Calcio/metabolismo , Caspasa 3 , Flavonoides , Interleucina-1 , Interleucina-6 , ARN Mensajero/genética , Ratas , Daño por Reperfusión/tratamiento farmacológico , Daño por Reperfusión/genética , Factor de Necrosis Tumoral alfa/genéticaRESUMEN
BACKGROUND: Klebsiella pneumoniae is a frequent nosocomial pathogen causing difficult-to-treat infections worldwide. The prevalence of Klebsiella pneumoniae carbapenemase (KPC)-producing Klebsiella pneumoniae (KPC-KP) is increasing in China. The aim of this study was to investigate the molecular epidemiology of KPC-KP in a nosocomial outbreak. METHODS: Fifty-four KPC-KP isolates were consecutively collected between November 2013 and August 2014 during a KPC-KP outbreak in a tertiary care hospital in Beijing, China. Antimicrobial susceptibility was determined by agar dilution. Carbapenemase, extended-spectrum ß-lactamase, 16S rRNA methylase, AmpC ß-lactamase, and plasmid-mediated quinolone resistance determinants were detected by PCR amplification. The genetic relatedness of isolates was analyzed by pulsed-field gel electrophoresis and multi-locus sequence typing. RESULTS: All isolates belonged to ST11 except one isolate which was identified as a new sequence type (ST2040). PFGE profile of genomic DNA revealed seven clusters, of which cluster A and C dominated the KPC-KP outbreak and cluster A was replaced by cluster C during the outbreak. PFGE of genomic DNA, S1-PFGE of plasmids, replicon typing, and drug resistant characteristics showed that clonal spread occurred during the outbreak. When compared with isolates within cluster A, all isolates in cluster C harbored rmtB and showed higher level of resistance to cefepime, amikacin, tobramycin, and tigecycline. CONCLUSION: We reported a nosocomial outbreak of KPC-KP with clonal replacement and a new sequence type (ST2040) of KP. High degree of awareness and surveillance of KPC-KP should be given to avoid potential outbreaks, especially in ICU wards.
Asunto(s)
Proteínas Bacterianas/metabolismo , Farmacorresistencia Bacteriana/genética , Infecciones por Klebsiella/microbiología , Klebsiella pneumoniae/patogenicidad , beta-Lactamasas/metabolismo , Proteínas Bacterianas/genética , China/epidemiología , Infección Hospitalaria/epidemiología , Brotes de Enfermedades , Farmacorresistencia Bacteriana/efectos de los fármacos , Electroforesis en Gel de Campo Pulsado , Hospitales , Humanos , Infecciones por Klebsiella/epidemiología , Infecciones por Klebsiella/mortalidad , Klebsiella pneumoniae/efectos de los fármacos , Klebsiella pneumoniae/aislamiento & purificación , Klebsiella pneumoniae/metabolismo , Metiltransferasas , Pruebas de Sensibilidad Microbiana , Epidemiología Molecular , Tipificación de Secuencias Multilocus , Plásmidos , ARN Ribosómico 16S , beta-Lactamasas/genéticaRESUMEN
E. moniliforme infections in humans have not been reported previously. We firstly described blood-stream infections caused by E. moniliforme in an elder woman with fractures of her left thigh. This study highlights the strategies to detect this anaerobic pathogen and the importance of investigating its molecular epidemiology in humans.
Asunto(s)
Bacteriemia/microbiología , Eubacterium/aislamiento & purificación , Fracturas del Fémur/complicaciones , Anciano , Femenino , Fracturas del Fémur/microbiología , HumanosRESUMEN
The clinical manifestations of Moraxella Atlantae infection were rarely described. Here we reported an elderly pneumonia patient with Moraxella Atlantae infection and the detailed clinical manifestations were firstly described. A bacterial automatic identification system in combination with phenotypic methods can be routinely used to identify this pathogen. If possible, 16S rDNA gene sequencing is also an alternative and effective method.
Asunto(s)
Bacteriemia/microbiología , Enfermedades de la Médula Ósea/microbiología , Moraxella/aislamiento & purificación , Infecciones por Moraxellaceae/microbiología , Neumonía Bacteriana/microbiología , Antibacterianos/uso terapéutico , Bacteriemia/diagnóstico , Bacteriemia/tratamiento farmacológico , Enfermedades de la Médula Ósea/diagnóstico , Enfermedades de la Médula Ósea/tratamiento farmacológico , Humanos , Masculino , Persona de Mediana Edad , Moraxella/clasificación , Moraxella/efectos de los fármacos , Moraxella/genética , Infecciones por Moraxellaceae/diagnóstico , Infecciones por Moraxellaceae/tratamiento farmacológico , Neumonía Bacteriana/diagnóstico , Neumonía Bacteriana/tratamiento farmacológico , Ribotipificación , Resultado del TratamientoRESUMEN
OBJECTIVES: Oral colonization of Candida could lead to later development of oropharyngeal candidiasis or candidemia among the immunocompromised patients. This study aims to describe the occurrence and risk factors of oral Candida colonization in patients with malignancies. MATERIALS AND METHODS: From October 2012 to March 2013, 78 patients with pulmonary cancer (group I), 101 patients with gastrointestinal tract tumor (group II), 79 patients with hematopoietic system malignant tumor (group III), and 101 healthy controls were consecutively recruited in a hospital in Beijing, China. The oral rinse samples were taken and Candida species were identified; the enzymes activities were tested. RESULTS: In total, 110 and 27 Candida strains were isolated from 91 patients and 26 controls, respectively. The oral colonization rate with Candida albicans in group III (12.7 %) was significant lower than that in group I (30.8 %), group II (33.7 %), and control group (25.7 %). The oral colonization rates with non-albicans Candida species in group I, group II, and group III were 15.4, 10.9, and 12.7 %, respectively, while only one non-albicans Candida strain was identified in control group. The non-albicans Candida species exhibited a lower virulence than C. albicans. Age was an independent risk factor for Candida colonization in patients with pulmonary cancer and digestive tract malignant tumor, "Teeth brush <1 time/day" was an independent risk factor for Candida colonization in patients with hematopoietic system tumor. CONCLUSIONS: The differences of risk factors for oral Candida colonization in patients with different cancers require different strategies for the prevention and control of Candida infection. CLINICAL RELEVANCE: Old aged patients with pulmonary cancer and digestive tract malignant tumor are high-risk population for Candida colonization. Increasing frequency of teeth brush might be helpful for preventing Candida colonization.
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Candidiasis Bucal/epidemiología , Candidiasis Bucal/microbiología , Neoplasias Gastrointestinales/complicaciones , Neoplasias Gastrointestinales/inmunología , Neoplasias Hematológicas/complicaciones , Neoplasias Hematológicas/inmunología , Neoplasias Pulmonares/complicaciones , Neoplasias Pulmonares/inmunología , Infecciones Oportunistas/epidemiología , Adulto , Antifúngicos/farmacología , Estudios de Casos y Controles , China/epidemiología , Femenino , Genotipo , Humanos , Huésped Inmunocomprometido , Masculino , Pruebas de Sensibilidad Microbiana , Persona de Mediana Edad , Estudios Prospectivos , Factores de Riesgo , Cepillado Dental , VirulenciaRESUMEN
Schizophyllum commune infections have been rarely reported. Here we reported a rare case of sinusitis in an acute myelocytic leukemia patient, who was co-infected by Escherichia coli, Stenotrophomonas maltophilia, and basidiomycetous fungi (Schizophyllum commune) in sinuses. Considering the in vitro and in vivo anti-fungal activity of voriconazole, it might be a good option to treat Schizophyllum commune infections when antifungal susceptibility testing is not available. When severe side effects occur, amphotericin B or itraconazole might be subsequent choice.
Asunto(s)
Escherichia coli/patogenicidad , Leucemia Mieloide Aguda/complicaciones , Micosis/complicaciones , Schizophyllum/patogenicidad , Sinusitis/microbiología , Adulto , Femenino , Humanos , Micosis/microbiología , Sinusitis/complicaciones , Sinusitis/diagnóstico por imagen , Tomografía Computarizada por Rayos XRESUMEN
Antibiotic-resistant opportunistic pathogens have become a serious concern in recent decades, as they are increasingly responsible for hospital-acquired infections. Here, we describe quinolone-resistant Delftia sp. strain 670, isolated from the sputum of a patient who died from severe pulmonary infection. The draft genome sequence of this strain was obtained by whole-genome shotgun sequencing, and was subjected to comparative genome analysis. Genome analysis revealed that one critical mutation (Ser83Ile in gyrA) might play a decisive role in quinolone resistance. The genome of Delftia sp. strain 670 contains both type II and type VI secretion systems, which were predicted to contribute to the virulence of the strain. Phylogenetic analysis, assimilation tests, and comparative genome analysis indicated that strain 670 differed from the four known Delftia species, suggesting this strain could represent a novel species. Although the study could not determine the strain 670 as the pathogen led to mortality, our findings also presented the pathogenic potential of Delftia species, and the increasing severity of antibiotic resistance among emerging opportunistic pathogens. The whole genome sequencing and comparative analysis improved our understanding of genome evolution in the genus Delftia, and provides the foundation for further study on drug resistance and virulence of Delftia strains.
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Antibacterianos/farmacología , Delftia/efectos de los fármacos , Delftia/genética , Farmacorresistencia Bacteriana , Genoma Bacteriano , Neumonía Bacteriana/microbiología , Quinolonas/farmacología , China , ADN Bacteriano/química , ADN Bacteriano/genética , Delftia/clasificación , Delftia/aislamiento & purificación , Resultado Fatal , Infecciones por Bacterias Gramnegativas/microbiología , Humanos , Datos de Secuencia Molecular , Filogenia , Análisis de Secuencia de ADN , Homología de Secuencia , Esputo/microbiologíaRESUMEN
BACKGROUND: Medical professionalism has been developing in the Peoples' Republic of China as one way to better address perennial and new challenges in healthcare in an ever-changing society. Among many recent developments in this area is promotion by the national Chinese Medical Doctor Association of the principles and values contained in the international document, "Medical Professionalism in the New Millennium: A Physician Charter." OBJECTIVE: To discover Chinese physicians' attitudes toward and understanding of medical professionalism. METHODOLOGY: The authors distributed a self-reporting questionnaire that included 34 statements and four case scenarios concerning the general principles of medical professionalism: the primacy of patients' welfare, respect for patients' autonomy, promotion of social justice, and professional self-regulation. The questionnaire included controversial issues such as the role of the family in decision making and reporting medical errors. A total of 2,966 practicing physicians, randomly selected from the Chinese Medical Association database, were surveyed, and 1,198 valid questionnaires were returned. Our sample covered 23 provinces and 51 cities throughout the Peoples' Republic of China. RESULTS: More than 80 percent of the physicians who responded agreed that the physician-patient relationship should be a relationship of trust founded on professional altruism, and that informed consent is necessary. More than 95 percent agreed that physicians should promote professional self-regulation as well as social justice. More than half agreed with the principle of the primacy of patients' welfare (62.8 percent), and that physicians have a responsibility to report medical errors and incompetent colleagues (51.0 percent). In certain cases, a great majority of Chinese physicians favored familism and paternalism. LIMITATIONS: The study does not include data on how Chinese physicians practice medical professionalism, or the perspectives of physicians working in smaller cities and in rural areas. CONCLUSIONS: Based on responses to the survey, Chinese physicians strongly support the majority of the fundamental principles and responsibilities of medical professionalism, including dedication, altruism, social justice, self-regulation, and informed consent. However, their support for the primacy of patients' welfare as a general principle, and the physician's responsibility to report medical errors and incompetent colleagues, is relatively low. To help advance medical professionalism in the People's Republic of China, professional development programs and medical ethics education should not only emphasize the general principles involved, but also formulate guidelines on how these principles can be carried out in practice.
Asunto(s)
Pueblo Asiatico/estadística & datos numéricos , Actitud del Personal de Salud , Comprensión , Relaciones Médico-Paciente/ética , Médicos/ética , Médicos/estadística & datos numéricos , Adulto , Altruismo , Discusiones Bioéticas , China/epidemiología , Características Culturales , Ética Médica , Femenino , Encuestas de Atención de la Salud , Humanos , Consentimiento Informado , Masculino , Persona de Mediana Edad , Paternalismo , Autonomía Personal , Médicos/psicología , Justicia Social , Estados UnidosRESUMEN
Stenotrophomonas maltophilia bacteriophage IME13 is a virulent phage with a large burst size, exceeding 3,000, much larger than that of any other stenotrophomonas phage reported before. It showed effective lysis of Stenotrophomonas maltophilia. Additionally, the phage IME13 developed at least three obviously different sizes of plaques when a single plaque was picked out and inoculated on a double-layer Luria broth agar plate with its host. Here we announce its complete genome and describe major findings from its annotation.
Asunto(s)
Bacteriófagos/genética , Genoma Viral , Stenotrophomonas maltophilia/virología , Secuencia de Bases , Mapeo Cromosómico , ADN Viral/genética , Datos de Secuencia Molecular , Análisis de Secuencia de ADN , Ensayo de Placa ViralRESUMEN
Tetralogy of Fallot (TOF) is the most common cyanotic congenital heart disease in newborns. ISL1 is a master transcription factor in second heart field development, whereas the roles of ISL1 gene promoter variants in TOF patients have not been genetically investigated. Total DNA extraction from 601 human subjects, including 308 TOF patients and 293 healthy controls, and Sanger sequencing were performed. Four variants (including one novel heterozygous variant) within the ISL1 gene promoter were only found in TOF patients. Functional analysis of DNA sequence variants was performed by using the dual-luciferase reporter assay and demonstrated that three of the four variants significantly decreased the transcriptional activity of ISL1 gene promoter in HL-1 cells (p < 0.05). Further, the online JASPAR database and electrophoretic mobility shift assay showed that the three variants affected the binding of transcription factors and altered ISL1 expression levels. In conclusion, the current study for the first time demonstrated that the variants identified from the ISL1 gene promoter region are likely involved in the development of TOF by affecting the transcriptional activity and altering the ISL1 expression level. Therefore, these findings may provide new insights into the molecular etiology and potential therapeutic strategy of TOF.
Asunto(s)
Cardiopatías Congénitas , Tetralogía de Fallot , Recién Nacido , Humanos , Tetralogía de Fallot/genética , Factores de Transcripción/genética , Cardiopatías Congénitas/genética , Regiones Promotoras Genéticas , HeterocigotoRESUMEN
Tetralogy of Fallot (TOF) is a common congenital heart malformation. Genetic variants in the CITED2 coding region are known to be significantly associated with cardiac malformation, but the role of variants in the CITED2 promoter region in the development of TOF remains unclear. In this study, we investigated CITED2 promoter variants in the DNA of 605 subjects, including 312 TOF patients and 293 unrelated healthy controls, by Sanger sequencing. We identified nine CITED2 gene promoter variants (including one novel heterozygous variant). Six were found only in patients with TOF and none in the control group. The transcriptional activity of the CITED2 gene promoter in mouse cardiomyocyte (HL-1) cells was significantly altered by the six variants (p < 0.05). The results of the electrophoretic mobility change assay and JASPAR database analysis showed that these variants generated or destroyed a series of possible transcription factor binding sites, resulting in changes in the CITED2 protein expression. We conclude that CITED2 promoter variants in TOF patients affect transcriptional activity and may be involved in the occurrence and progression of TOF. These findings may provide new insights into molecular pathogenesis and potential therapeutic insights in patients with TOF.
Asunto(s)
Cardiopatías Congénitas , Tetralogía de Fallot , Ratones , Animales , Tetralogía de Fallot/genética , Cardiopatías Congénitas/genética , Regiones Promotoras Genéticas , Proteínas Represoras/genética , Transactivadores/genéticaRESUMEN
Atrial septal defect (ASD) is one of the most common forms of congenital heart disease (CHD). Genetic variants in the coding region of the CITED2 gene are known to be significantly correlated with CHD, but the role of variants in the promoter region of CITED2 is unknown. We investigated variants in the promoter of the CITED2 gene in 625 subjects (332 ASD and 293 healthy controls) through Sanger sequencing. Four variants in the CITED2 gene promoter were found only in eight ASD patients with zero occurrence in the control subjects (one case of g.4078A>C(rs1165649373), one case of g.4240C>A(rs1235857801), four cases of g.4935C>T(rs111470468), two cases of g.5027C>T(rs112831934)). Cellular functional analysis showed that these four variants significantly changed the transcriptional activity of the CITED2 gene promoter in HEK-293 and HL-1 cells. Electrophoretic mobility change assay results and JASPAR database analysis demonstrated that these variants created or destroyed a series of possible transcription factor binding sites, resulting in changes in the expression of CITED2 protein. We conclude that the variants of CITED2 promoter in ASD patients affect the transcriptional activity and are likely involved in the occurrence and development of ASD. These findings provide new perspectives on the pathogenesis and potential therapeutic insights of ASD.
RESUMEN
OBJECTIVE: To study Fusarium solani infection as a complication in patients after allogeneic hematopoietic stem cell transplantation and to discuss the diagnosis and appropriate therapy. METHODS: Symptoms, physical examination, laboratory tests, computed tomographic (CT) scans, treatments and outcomes of Fusarium solani infection in a patient with acute myeloid leukemia after allogeneic hematopoietic stem cell transplantation were retrospectively analyzed, and related literatures reviewed. RESULTS: The patient developed pulmonary infiltration and systemic multiple subcutaneous masses after allogeneic hematopoietic stem cell transplantation. Tissue biopsy smear showed a large number of hyphae and spores, and fungal culture grew Fusarium solani. The subcutaneous masses were incised and drained, while amphotericin B and voriconazole were administered, with granulocyte colony-stimulating factor (G-CSF) and granulocyte-macrophage colony-stimulating factor (GM-CSF) for hematopoietic recovery. The patient was discharge after full recovery. CONCLUSION: Fusarium solani infection is a rare but fatal complication after allogeneic hematopoietic stem cell transplantation. Once the skin lesions or subcutaneous masses developed, tissue smear and culture should be done as soon as possible. Early diagnosis and effective treatment to recovery of the patient after allogeneic hematopoietic stem cell transplant. Moreover, the recovery of adequate neutrophil levels is the most important factor in the resolution of fusarial infection.
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Fusariosis/diagnóstico , Fusarium/patogenicidad , Trasplante de Células Madre Hematopoyéticas , Enfermedades de la Piel/microbiología , Fusariosis/tratamiento farmacológico , Humanos , Masculino , Persona de Mediana Edad , Enfermedades de la Piel/diagnóstico , Enfermedades de la Piel/tratamiento farmacológico , Trasplante HomólogoRESUMEN
Acinetobacter baumannii is an important nosocomial pathogen in hospital-acquired infections, and carbapenem resistance has been increasingly observed worldwide. Oxacillinase production by blaOXA-23 is a predominant and prevalent carbapenem resistance mechanism of A. baumannii, especially in China. Rapid and specific detection of blaOXA-23 may offer valuable insight for administration of directed antimicrobial therapy. In this study, we aimed to develop a loop-mediated isothermal amplification (LAMP)-based method for identifying carbapenem-resistant A. baumannii (CRAB) harboring the blaOXA-23 gene. High-specificity primers for screening blaOXA-23 were designed and synthesized, and the LAMP reactions were performed. Clinical A. baumannii strains isolated from the Former 307th Hospital of People's Liberation Army were used to determine the sensitivity and specificity of this method compared with those of phenotypic antimicrobial susceptibility testing and the traditional PCR method. Multilocus sequence typing (MLST) was performed to investigate the epidemiology of the A. baumannii bacterial population. Compared with antimicrobial susceptibility testing, the sensitivity and specificity of LAMP in detecting blaOXA-23 were 88.4% and 97.7%, respectively. However, the LAMP method is much simpler and less time-consuming (within 60 minutes) than conventional PCR and phenotypic susceptibility testing. The 113 isolates could be clustered into 30 sequence types, and most strains (83/113) belonged to clonal complex (CC) 92, which is also the dominant CC in China. The LAMP-based method detected blaOXA-23 in a simpler manner and could provide rapid results for identifying CRAB. Consequently, blaOXA-23 may serve as a surrogate marker for the presence of CRAB in patients with serious infections in clinical practice.
Asunto(s)
Acinetobacter baumannii/genética , Antibacterianos/farmacología , Proteínas Bacterianas/genética , Farmacorresistencia Bacteriana Múltiple/genética , Técnicas de Diagnóstico Molecular/métodos , Técnicas de Amplificación de Ácido Nucleico/métodos , beta-Lactamasas/genética , Acinetobacter baumannii/efectos de los fármacos , Farmacorresistencia Bacteriana Múltiple/efectos de los fármacos , Pruebas de Sensibilidad Microbiana , Tipificación de Secuencias MultilocusRESUMEN
We have previously described an automated system (ECIS/taxis) for measuring chemotactic movement of Dictyostelium amoebae in a folic acid gradient [Hadjout, N., Laevsky, G., Knecht, D.A. and Lynes, M.A., 2001. Automated real-time measurement of chemotactic cell motility. Biotechniques 31, 1130-1138.]. In the ECIS/taxis system, cells migrate in an under-agarose environment, and their position is monitored by determining the impedance change caused by cells crawling onto the surface of an electrode. In this report, we show that chemotaxis of primary and immortalized leukocytes in response to complement (C5a) could be measured using the ECIS/taxis system. Several modifications to the design of the target electrode were tested, and a linear electrode perpendicular to the direction of movement was found to increase the sensitivity and reliability of the assay. Using the optimized ECIS/taxis assay, the dose response of neutrophils and WBC 265-9C cells was established and compared to the Boyden chamber assay. The ECIS/taxis assay system can be used to compare the movement of different cell types, to assess the effect of complex chemotactic gradients, or to determine the effects of pharmaceuticals on chemotactic motility.
Asunto(s)
Células de la Médula Ósea/inmunología , Quimiotaxis de Leucocito , Complemento C5a/inmunología , Sistemas de Computación , Neutrófilos/inmunología , Animales , Células de la Médula Ósea/efectos de los fármacos , Línea Celular , Movimiento Celular , Células Cultivadas , Complemento C5a/farmacología , Relación Dosis-Respuesta a Droga , Electrodos , Humanos , Ratones , Ratones Endogámicos C57BL , Neutrófilos/efectos de los fármacos , Factores de TiempoRESUMEN
OBJECTIVES: Heteroresistance is a phenomenon in which there are various responses to antibiotics from bacterial cells within the same population. Here, we isolated and characterised an imipenem heteroresistant Acinetobacter baumannii strain (HRAB-85). METHODS: The genome of strain HRAB-85 was completely sequenced and analysed to understand its antibiotic resistance mechanisms. Population analysis and multilocus sequence typing were performed. RESULTS: Subpopulations grew in the presence of imipenem at concentrations of up to 64µg/mL, and the strain was found to belong to ST208. The total length of strain HRAB-85 was 4,098,585bp with a GC content of 39.98%. The genome harboured at least four insertion sequences: the common ISAba1, ISAba22, ISAba24, and newly reported ISAba26. Additionally, 19 antibiotic-resistance genes against eight classes of antimicrobial agents were found, and 11 genomic islands (GIs) were identified. Among them, GI3, GI10, and GI11 contained many ISs and antibiotic-resistance determinants. CONCLUSIONS: The existence of imipenem heteroresistant phenotypes in A. baumannii was substantiated in this hospital, and imipenem pressure, which could induce imipenem-heteroresistant subpopulations, may select for highly resistant strains. The complete genome sequencing and bioinformatics analysis of HRAB-85 could improve our understanding of the epidemiology and resistance mechanisms of carbapenem-heteroresistant A. baumannii.