RESUMO
IMPORTANCE: HPV DNA screening is an effective approach for the prevention of cervical cancer. The novel real-time recombinase polymerase amplification-based HPV detection systems we developed constitute an improvement over the HPV detection methods currently used in clinical practice and should help to extend cervical cancer screening in the future, particularly in point-of-care test settings.
Assuntos
Infecções por Papillomavirus , Neoplasias do Colo do Útero , Feminino , Humanos , Papillomavirus Humano 16/genética , Papillomavirus Humano 18/genética , Neoplasias do Colo do Útero/diagnóstico , Recombinases , Infecções por Papillomavirus/diagnóstico , Detecção Precoce de Câncer/métodos , DNA Viral/genéticaRESUMO
Monkeypox is a zoonotic disease caused by monkeypox virus (MPXV), in which outbreaks mainly occurred in West and Central Africa, with only sporadic spillovers to countries outside Africa due to international travel or close contact with wildlife. During May 2022, multiple countries in Europe, North and South America, Australia, Asia, and Africa reported near-simultaneous outbreaks of MPXV, the first time that patient clusters were reported over such a large geographical area. Cases have no known epidemiological links to MPXV-endemic countries in West or Central Africa. Real-time PCR is currently the gold standard for MPXV diagnostics, but it requires trained laboratory personnel and specialized equipment, and results can only be obtained after several hours. A rapid and simple-to-operate point-of-care diagnostic test for MPXV is crucial for limiting its spread and controlling outbreaks. Here, three recombinase-based isothermal amplification assays (RPA/RAA) for the rapid detection of MPXV isolates were developed. These three assays target the MPXV G2R gene, and the limit of detection for these systems is approximately 100 copies of DNA per reaction. The assays were found to be specific and non-cross reactive against other pox viruses, such as vaccinia virus, and the results can be visualized within 20-30 min. The assays were validated with DNA extracted from 19 clinical samples from suspected or confirmed MPXV patients from Central Africa, and found to be consistent with findings from traditional qPCR. These results provide a solid platform for the early diagnosis of potential MPXV cases, and will help with the control and prevention of current and future outbreaks.
Assuntos
Monkeypox virus , Mpox , Humanos , Monkeypox virus/genética , Recombinases , Mpox/diagnóstico , Mpox/epidemiologia , Reação em Cadeia da Polimerase em Tempo Real , Europa (Continente) , Técnicas de Amplificação de Ácido Nucleico/métodos , Sensibilidade e EspecificidadeRESUMO
Bladder cancer is a common tumor type of the urinary system, which has high levels of morbidity and mortality. The firstline treatment is cisplatinbased combination chemotherapy, but a significant proportion of patients relapse due to the development of drug resistance. Therapyinduced senescence can act as a 'backup' response to chemotherapy in cancer types that are resistant to apoptosisbased anticancer therapies. The circadian clock serves an important role in drug resistance and cellular senescence. The aim of the present study was to investigate the regulatory effect of the circadian clock on paclitaxel (PTX)induced senescence in cisplatinresistant bladder cancer cells. Cisplatinresistant bladder cancer cells were established via longterm cisplatin incubation. PTX induced apparent senescence in bladder cancer cells as demonstrated via SAßGal staining, but this was not observed in the cisplatinresistant cells. The cisplatinresistant cells entered into a quiescent state with prolonged circadian rhythm under acute PTX stress. It was identified that the circadian protein cryptochrome1 (CRY1) accumulated in these quiescent cisplatinresistant cells, and that CRY1 knockdown restored PTXinduced senescence. Mechanistically, CRY1 promoted p53 degradation via increasing the binding of p53 with its ubiquitin E3 ligase MDM2 protooncogene. These data suggested that the accumulated CRY1 in cisplatinresistant cells could prevent PTXinduced senescence by promoting p53 degradation.