RESUMO
BACKGROUND: Colorectal cancer (CRC) ranks as the third most common malignancies in the world, and periodic examination of the patient is advantageous in reducing the mortality of CRC. The first blood-based Septin9 gene methylation assay which recognized by the US FDA for CRC examination was Epi proColon. However, this assay was not broadly applied in the current clinical guideline because of its relatively lower sensitivity in the detection of early-stage CRC. METHODS: This study aimed at developing a new multiplex Septin9 methylation assay (ColonUSK) which simultaneously evaluates two CpG-rich subregions in the promoter of the Septin9 gene and an internal control in a single reaction. ColonUSK proved increased sensitivity, with a detection limit as low as 12pg of the positive DNA compared with the Septin9 assay targeting one CpG-rich subregion. 1366 subjects were prospectively recruited from four comprehensive hospitals in China in an opportunistic screening study for assessing its value in CRC detection. Blind testing was developed to evaluate ColonUSK in comparison with clinical examination using clinical gold standard such as colonoscopy. RESULTS: The assay demonstrates clinical sensitivity for diagnosing colorectal cancer (CRC) and advanced adenoma at rates of 77.34% and 25.26%, respectively. Furthermore, ColonUSK exhibits a high degree of specificity for non-CRC cases (95.95%) clinically. Significantly, the detection rate of cases in high-grade intraepithelial neoplasia increased to 54.29%. The value for the assay in the Kappa test was 0.76, showing a high degree of consistency between ColonUSK and clinical gold standard. CONCLUSIONS: ColonUSK indicated moderate diagnostic value and could become a non-invasive detection way for CRC. The implementation of the ColonUSK assay has the capacity to markedly enhance CRC screening practices.
Assuntos
Neoplasias Colorretais , Metilação de DNA , Detecção Precoce de Câncer , Septinas , Humanos , Septinas/genética , Neoplasias Colorretais/diagnóstico , Neoplasias Colorretais/genética , Masculino , Feminino , Pessoa de Meia-Idade , Detecção Precoce de Câncer/métodos , Idoso , Regiões Promotoras Genéticas , Sensibilidade e Especificidade , Biomarcadores Tumorais/genética , Ilhas de CpG , Estadiamento de Neoplasias , Adulto , Estudos Prospectivos , Gradação de TumoresRESUMO
Base editors, including dual base editors, are innovative techniques for efficient base conversions in genomic DNA. However, the low efficiency of A-to-G base conversion at positions proximal to the protospacer adjacent motif (PAM) and the A/C simultaneous conversion of the dual base editor hinder their broad applications. In this study, through fusion of ABE8e with Rad51 DNA-binding domain, we generate a hyperactive ABE (hyABE) which offers improved A-to-G editing efficiency at the region (A10-A15) proximal to the PAM, with 1.2- to 7-fold improvement compared to ABE8e. Similarly, we develop optimized dual base editors (eA&C-BEmax and hyA&C-BEmax) with markedly improved simultaneous A/C conversion efficiency (1.2-fold and 1.5-fold improvement, respectively) compared to A&C-BEmax in human cells. Moreover, these optimized base editors catalyze efficiently nucleotide conversions in zebrafish embryos to mirror human syndrome or in human cells to potentially treat genetic diseases, indicating their great potential in broad applications for disease modeling and gene therapy.
Assuntos
Adenina , Peixe-Zebra , Humanos , Animais , Nucleotídeos , Catálise , Terapia GenéticaRESUMO
Depression is a serious public health problem and an important factor leading to disease-related disability. Influenced by many factors, such as psychological, hormonal, and genetic factors, the incidence rate of depression in females is approximately two times that in males. However, in preclinical neuroscience research, the selection of the animals' sex for use in depression models has been controversial. At present, in most preclinical studies, the animals generally chosen in depression models have been male rodents rather than female rodents. It remains doubtful whether the data obtained from male animals can be generalized to female animals. The performance of female animals in preclinical studies of depression has been inconclusive. Based on a review of a large number of original studies in the PubMed database, it was found that although male rodents are more commonly used in the study of depression, the use of female animals also shows good modeling of depression and has its advantages. The influence of the animals' sex in the chronic unpredictable mild stress (CUMS) model needs further research.
Assuntos
Antidepressivos , Depressão , Animais , Antidepressivos/farmacologia , Comportamento Animal , Depressão/etiologia , Modelos Animais de Doenças , Feminino , Hipocampo , Masculino , Estresse PsicológicoRESUMO
Despite tremendous success of molecular targeted therapy together with immunotherapy, only a small subset of patients can benefit from them. Chemotherapy remains the mainstay treatment for most of tumors including non-small cell lung cancer (NSCLC); however, non-selective adverse effects on healthy tissues and secondary resistance are the main obstacles. Meanwhile, the quiescent or dormant cancer stem-like cells (CSLCs) are resistant to antimitotic chemoradiotherapy. Complete remission can only be realized when both proliferative cancer cells and quiescent cancer stem cells are targeted. In the present research, we constructed a cooperatively combating conjugate (DTX-P7) composed of docetaxel (DTX) and a heptapeptide (P7), which specifically binds to cell surface Hsp90, and assessed the anti-tumor effects of DTX-P7 on non-small cell lung cancer. DTX-P7 preferentially suppressed tumor growth compared with DTX in vivo with a favorable distribution to tumor tissues and long circulation half-life. Furthermore, we revealed a distinctive mechanism whereby DTX-P7 induced unfolded protein response and eventually promoted apoptosis. More importantly, we found that DTX-P7 promoted cell cycle reentry of slow-proliferating CSLCs and subsequently killed them, exhibiting a "proliferate to kill" pattern. Collecitvely, by force of active targeting delivery of DTX via membrane-bound Hsp90, DTX-P7 induces unfolded protein response and subsequent apoptosis by degrading Hsp90, meanwhile awakens and kills the dormant cancer stem cells. Thus, DTX-P7 deserves further development as a promising anticancer therapeutic for treatment of various membrane-harboring Hsp90 cancer types.
Assuntos
Antineoplásicos , Carcinoma Pulmonar de Células não Pequenas , Neoplasias Pulmonares , Antineoplásicos/farmacologia , Antineoplásicos/uso terapêutico , Carcinoma Pulmonar de Células não Pequenas/tratamento farmacológico , Carcinoma Pulmonar de Células não Pequenas/patologia , Linhagem Celular Tumoral , Docetaxel/química , Docetaxel/farmacologia , Portadores de Fármacos/química , Humanos , Neoplasias Pulmonares/tratamento farmacológico , Neoplasias Pulmonares/patologia , Peptídeos/química , Peptídeos/farmacologia , Peptídeos/uso terapêuticoRESUMO
Myeloid-derived suppressor cells (MDSCs) play an important role in tumor immune escape. Recent studies have shown that MDSCs contribute to tumor progression under psychological stress, but the underlying mechanism of MDSCs mobilization and recruitment remains largely unknown. In the present study, a chronic restraint stress paradigm was applied to the H22 hepatocellular carcinoma (HCC) bearing mice to mimic the psychological stress. We observed that chronic restraint stress significantly promoted HCC growth, as well as the mobilization of MDSCs to spleen and tumor sites from bone marrow. Meanwhile, chronic restraint stress enhanced the expression of C-X-C motif chemokine receptor 2 (CXCR2) and pErk1/2 in bone marrow MDSCs, together with elevated chemokine (C-X-C motif) ligand 5 (CXCL5) expression in tumor tissues. In vitro, the treatments of MDSCs with epinephrine (EPI) and norepinephrine (NE) but not corticosterone (CORT)-treated H22 conditioned medium obviously inhibited T-cell proliferation, as well as enhanced CXCR2 expression and extracellular signal-regulated kinase (Erk) phosphorylation. In vivo, ß-adrenergic blockade with propranolol almost completely reversed the accelerated tumor growth induced by chronic restraint stress and inactivated CXCL5-CXCR2-Erk signaling pathway. Our findings support the crucial role of ß-adrenergic signaling cascade in the mobilization and recruitment of MDSCs under chronic restraint stress.
Assuntos
Antagonistas Adrenérgicos beta/administração & dosagem , Carcinoma Hepatocelular/tratamento farmacológico , Neoplasias Hepáticas/tratamento farmacológico , Células Supressoras Mieloides/metabolismo , Propranolol/administração & dosagem , Estresse Psicológico/complicações , Antagonistas Adrenérgicos beta/farmacologia , Animais , Carcinoma Hepatocelular/metabolismo , Carcinoma Hepatocelular/patologia , Linhagem Celular Tumoral , Quimiocina CXCL5 , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Neoplasias Hepáticas/metabolismo , Neoplasias Hepáticas/patologia , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Masculino , Camundongos , Células Supressoras Mieloides/efeitos dos fármacos , Transplante de Neoplasias , Propranolol/farmacologia , Receptores de Interleucina-8B , Baço/imunologia , Estresse Psicológico/etiologia , Estresse Psicológico/metabolismoRESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: ANNAO tablets derive from Chinese classical prescriptions of Angong Niuhuang Pills with modified compositions, which have been singly or combined used for stoke associated neurological disorders. However the underlying mechanism is not yet well-defined, the present study investigated its anti-ischemic effects in rat middle cerebral artery occlusion (MCAO) model and focused on mitochondrial quality control. MATERIALS AND METHODS: Rats were subjected to 2â¯h of brain ischemia followed by 1 day or up to 7 days of reperfusion. Vehicle, ANNAO tablets or Edaravone were given at 1h after the start of reperfusion for 1 day or successive 7 days in MCAO rats. For the behavior assessment, Longa test and modified Neurological Severity Scores (m NSS) test were performed. Following the behavioral assessment, we assessed the protein expressions related to mitochondrial function. Moreover, we also assessed the neuroprotective effects of ANNAO tablets by immunohistochemical analysis. RESULTS: Compared with sham rats, ANNAO tablets improved the behavioral performance and decreased the infarction volume in the MCAO rats. Western blotting results showed that ANNAO tablets altered the expression level of multiple proteins related to mitochondrial function, elevated the ratio of Bcl-2/Bax and inhibited the apoptosis. Additionally, ANNAO tablets increased the number of NeuN positive neurons. CONCLUSIONS: The obtained data demonstrated that ANNAO tablets exhibited an obvious anti-cerebral ischemia-reperfusion effect, which could be attributed to the improvement of mitochondrial quality control.
Assuntos
Infarto da Artéria Cerebral Média/tratamento farmacológico , Mitocôndrias/efeitos dos fármacos , Fármacos Neuroprotetores/farmacologia , Fármacos Neuroprotetores/uso terapêutico , Traumatismo por Reperfusão/tratamento farmacológico , Animais , Apoptose/efeitos dos fármacos , Masculino , Mitocôndrias/fisiologia , Dinâmica Mitocondrial/efeitos dos fármacos , Mitofagia/efeitos dos fármacos , Ratos Sprague-Dawley , ComprimidosRESUMO
BACKGROUND: Serum CA-125 has been used as a biomarker of gynecological tumors. In this study, we investigated the CA-125 levels in cervical and vaginal secretions from Chinese patients with endometrial polyps, hyperplasia and carcinoma in comparison with those in endometrium and serum. MATERIAL/METHODS: An electro-chemiluminescent immunoassay was utilized to determine the levels of CA-125 in 51 healthy Chinese women and 97 patients with polyps, hyperplasia or endometrial cancer. An immunohistochemistry method was used to detect endometrial CA-125 expression in 242 subjects. RESULTS: Our study demonstrated that serum CA-125 levels were much lower than those in cervical and vaginal secretions in healthy and diseased women. The levels of CA-125 in serum, and cervical and vaginal secretions were significantly increased in complex hyperplasia and endometrial cancer. The increase of CA-125 content in serum, cervical and vaginal secretions was lesser significant in grade 3 cancer than that in grade 1 and 2 cancer. Generally, serum CA-125 levels correlated with those in cervical and vaginal secretions and CA-125 content in cervical secretion correlated with that in vaginal secretion. There was only a weak CA-125 expression in normal endometrium and simple endometrial hyperplasia. There was a significant difference in CA-125 expression among patients with pathological grade 1, 2 and 3 of endometrial carcinoma. CONCLUSIONS: Endo.metrial CA-125 expression together with its levels in the serum and cervical and vaginal secretions can be used as a potential biomarker in the diagnosis of precancerous diseases and endometrial carcinoma.