RESUMO
Here, we describe pathological events potentially involved in the disease pathogenesis of Alexander disease (AxD). This is a primary genetic disorder of astrocyte caused by dominant gain-of-function mutations in the gene coding for an intermediate filament protein glial fibrillary acidic protein (GFAP). Pathologically, this disease is characterized by the upregulation of GFAP and its accumulation as Rosenthal fibers. Although the genetic basis linking GFAP mutations with Alexander disease has been firmly established, the initiating events that promote GFAP accumulation and the role of Rosenthal fibers (RFs) in the disease process remain unknown. Here, we investigate the hypothesis that disease-associated mutations promote GFAP aggregation through aberrant posttranslational modifications. We found high molecular weight GFAP species in the RFs of AxD brains, indicating abnormal GFAP crosslinking as a prominent pathological feature of this disease. In vitro and cell-based studies demonstrate that cystine-generating mutations promote GFAP crosslinking by cysteine-dependent oxidation, resulting in defective GFAP assembly and decreased filament solubility. Moreover, we found GFAP was ubiquitinated in RFs of AxD patients and rodent models, supporting this modification as a critical factor linked to GFAP aggregation. Finally, we found that arginine could increase the solubility of aggregation-prone mutant GFAP by decreasing its ubiquitination and aggregation. Our study suggests a series of pathogenic events leading to AxD, involving interplay between GFAP aggregation and abnormal modifications by GFAP ubiquitination and oxidation. More important, our findings provide a basis for investigating new strategies to treat AxD by targeting abnormal GFAP modifications.
Assuntos
Doença de Alexander , Proteína Glial Fibrilar Ácida , Ubiquitinação , Doença de Alexander/metabolismo , Doença de Alexander/genética , Doença de Alexander/patologia , Proteína Glial Fibrilar Ácida/metabolismo , Proteína Glial Fibrilar Ácida/genética , Humanos , Animais , Mutação , Camundongos , Astrócitos/metabolismo , Astrócitos/patologia , Encéfalo/metabolismo , Encéfalo/patologia , Processamento de Proteína Pós-Traducional , Ratos , Masculino , Feminino , Agregação Patológica de Proteínas/metabolismo , Agregação Patológica de Proteínas/genética , Agregação Patológica de Proteínas/patologiaRESUMO
OBJECTIVES: To investigate the risk factors for bronchopulmonary dysplasia (BPD) in twin preterm infants with a gestational age of <34 weeks, and to provide a basis for early identification of BPD in twin preterm infants in clinical practice. METHODS: A retrospective analysis was performed for the twin preterm infants with a gestational age of <34 weeks who were admitted to 22 hospitals nationwide from January 2018 to December 2020. According to their conditions, they were divided into group A (both twins had BPD), group B (only one twin had BPD), and group C (neither twin had BPD). The risk factors for BPD in twin preterm infants were analyzed. Further analysis was conducted on group B to investigate the postnatal risk factors for BPD within twins. RESULTS: A total of 904 pairs of twins with a gestational age of <34 weeks were included in this study. The multivariate logistic regression analysis showed that compared with group C, birth weight discordance of >25% between the twins was an independent risk factor for BPD in one of the twins (OR=3.370, 95%CI: 1.500-7.568, P<0.05), and high gestational age at birth was a protective factor against BPD (P<0.05). The conditional logistic regression analysis of group B showed that small-for-gestational-age (SGA) birth was an independent risk factor for BPD in individual twins (OR=5.017, 95%CI: 1.040-24.190, P<0.05). CONCLUSIONS: The development of BPD in twin preterm infants is associated with gestational age, birth weight discordance between the twins, and SGA birth.
Assuntos
Displasia Broncopulmonar , Recém-Nascido Prematuro , Gêmeos , Humanos , Displasia Broncopulmonar/etiologia , Displasia Broncopulmonar/epidemiologia , Fatores de Risco , Recém-Nascido , Feminino , Estudos Retrospectivos , Masculino , Idade Gestacional , Peso ao Nascer , Modelos LogísticosRESUMO
Viruses employ multiple strategies to inhibit host mRNA nuclear export. Distinct to the generally nonselective inhibition mechanisms, ORF10 from gammaherpesviruses inhibits mRNA export in a transcript-selective manner by interacting with Rae1 (RNA export 1) and Nup98 (nucleoporin 98). We now report the structure of ORF10 from MHV-68 (murine gammaherpesvirus 68) bound to the Rae1-Nup98 heterodimer, thereby revealing detailed intermolecular interactions. Structural and functional assays highlight that two highly conserved residues of ORF10, L60 and M413, play critical roles in both complex assembly and mRNA export inhibition. Interestingly, although ORF10 occupies the RNA-binding groove of Rae1-Nup98, the ORF10-Rae1-Nup98 ternary complex still maintains a comparable RNA-binding ability due to the ORF10-RNA direct interaction. Moreover, mutations on the RNA-binding surface of ORF10 disrupt its function of mRNA export inhibition. Our work demonstrates the molecular mechanism of ORF10-mediated selective inhibition and provides insights into the functions of Rae1-Nup98 in regulating host mRNA export.
Assuntos
Transporte de RNA/fisiologia , RNA Mensageiro/metabolismo , Transativadores/metabolismo , Animais , Células HEK293 , Interações Hospedeiro-Patógeno , Humanos , Camundongos , Proteínas Associadas à Matriz Nuclear/química , Proteínas Associadas à Matriz Nuclear/metabolismo , Complexo de Proteínas Formadoras de Poros Nucleares/química , Complexo de Proteínas Formadoras de Poros Nucleares/metabolismo , Proteínas de Transporte Nucleocitoplasmático/química , Proteínas de Transporte Nucleocitoplasmático/metabolismo , RNA Mensageiro/química , Células Sf9 , Transativadores/químicaRESUMO
Alexander disease (AxD) caused by mutations in the coding region of GFAP is a neurodegenerative disease characterized by astrocyte dysfunction, GFAP aggregation, and Rosenthal fiber accumulation. Although how GFAP mutations cause disease is not fully understood, Rosenthal fibers could be induced by forced overexpression of human GFAP and this could be lethal in mice implicate that an increase in GFAP levels is central to AxD pathogenesis. Our recent studies demonstrated that intronic GFAP mutations cause disease by altering GFAP splicing, suggesting that an increase in GFAP isoform expression could lead to protein aggregation and astrocyte dysfunction that typify AxD. Here we test this hypothesis by establishing primary astrocyte cultures from transgenic mice overexpressing human GFAP. We found that GFAP-δ and GFAP-κ were disproportionately increased in transgenic astrocytes and both were enriched in Rosenthal fibers of human AxD brains. In vitro assembly studies showed that while the major isoform GFAP-α self-assembled into typical 10-nm filaments, minor isoforms including GFAP-δ, -κ, and -λ were assembly-compromised and aggregation prone. Lentiviral transduction showed that expression of these minor GFAP isoforms decreased filament solubility and increased GFAP stability, leading to the formation of Rosenthal fibers-like aggregates that also disrupted the endogenous intermediate filament networks. The aggregate-bearing astrocytes lost their normal morphology and glutamate buffering capacity, which had a toxic effect on neighboring neurons. In conclusion, our findings provide evidence that links elevated GFAP isoform expression with GFAP aggregation and impaired glutamate transport, and suggest a potential non-cell-autonomous mechanism underlying neurodegeneration through astrocyte dysfunction.
Assuntos
Doença de Alexander/patologia , Astrócitos/patologia , Proteína Glial Fibrilar Ácida/química , Proteína Glial Fibrilar Ácida/metabolismo , Ácido Glutâmico/metabolismo , Mutação , Agregados Proteicos , Doença de Alexander/metabolismo , Animais , Astrócitos/metabolismo , Humanos , Camundongos , Camundongos Transgênicos , Conformação Proteica , Isoformas de ProteínasRESUMO
PURPOSE: Studies have reported a strong link between asthma and panic disorder. We conducted a 17-year community-based large cohort study to examine the relationship between asthma, early smoking initiation, and panic disorder during adolescence and early adulthood. METHODS: A total of 162,766 participants aged 11-16 years were categorized into asthma and nonasthma groups at baseline and compared within the observation period. Covariates during late childhood or adolescence included parental education, cigarette smoking by family members of participants, and participant's gender, age, alcohol consumption, smoking, and exercise habits. Data for urbanicity, prednisone use, allergic comorbidity, and Charlson comorbidity index were acquired from the National Health Insurance Research Database. The Cox proportional-hazards model was used to evaluate the association between asthma and panic disorder. RESULTS: Our findings revealed that asthma increased the risk of panic disorder after adjustment for key confounders in the Cox proportional hazard regression model (adjusted HR: 1.70, 95% CI 1.28-2.26). Hospitalizations or visits to the emergency department for asthma exhibited a dose-response effect on the panic disorder (adjusted HR: 2.07, 95% CI 1.30-3.29). Patients with asthma with onset before 20 years of age who smoked during late childhood or adolescence had the greatest risk for panic disorder (adjusted HR: 4.95, 95% CI 1.23-19.90). CONCLUSIONS: Patients newly diagnosed with asthma had a 1.7-times higher risk of developing panic disorder. Smoking during late childhood or adolescence increased the risk for developing the panic disorder in patients with asthma.
Assuntos
Asma , Transtorno de Pânico , Adolescente , Adulto , Asma/epidemiologia , Criança , Estudos de Coortes , Humanos , Transtorno de Pânico/diagnóstico , Transtorno de Pânico/epidemiologia , Fatores de Risco , Fumar/efeitos adversos , Fumar/epidemiologia , Adulto JovemRESUMO
Bisphenol A (BPA) is becoming a potential environmental toxicity factor. However, BPA's effect and function mechanism on maize roots remain unknown. Here, we investigated characters of root growth of maize seedlings exposed to BPA for 8 d and without BPA for 3 d, and a series of indicators on reactive oxygen homeostasis and nitrogen assimilation were measured. High-dose BPA(15 and 50 mg·L-1) suppressed the root growth and caused increased contents of O2Ë-, H2O2 and MDA in maize seedling roots. The disturbed ROS homeostasis resulted from the change of antioxidant enzymes, including the increase of APX, GPX, and CAT, and decrease of SOD and POD, and a decrease of antioxidant substance GSH. Meanwhile, High-dose BPA caused a decrease in the soluble protein content, nitrate reductase (NR), glutamate dehydrogenase (GDH), and glutamine oxoglutarate aminotransferase (GOGAT) under the BPA processing phase and recovery period. The low-dose BPAï¼1.5 and 5 mg·L-1ï¼significantly promoted root growth of maize seedlings and maintained the ROS homeostasis through antioxidant enzyme APX and GPX eliminating redundant ROS. Our results showed that BPA could cause a dual effect on the root growth of maize seedlings, that is, promotion of low-dose and inhibition of high-dose, through ROS homeostasis and nitrogen assimilation in Zea mays.
Assuntos
Plântula , Zea mays , Antioxidantes , Nitrogênio , Peróxido de Hidrogênio , Espécies Reativas de OxigênioRESUMO
OBJECTIVES: To investigate the clinical treatment outcomes and the changes of the outcomes over time in extremely preterm twins in Guangdong Province, China. METHODS: A retrospective analysis was performed for 269 pairs of extremely preterm twins with a gestational age of <28 weeks who were admitted to the department of neonatology in 26 grade A tertiary hospitals in Guangdong Province from January 2008 to December 2017. According to the admission time, they were divided into two groups: 2008-2012 and 2013-2017. Besides, each pair of twins was divided into the heavier infant and the lighter infant subgroups according to birth weight. The perinatal data of mothers and hospitalization data of neonates were collected. The survival rate of twins and the incidence rate of complications were compared between the 2008-2012 and 2013-2017 groups. RESULTS: Compared with the 2008-2012 group, the 2013-2017 group (both the heavier infant and lighter infant subgroups) had lower incidence rates of severe asphyxia and smaller head circumference at birth (P<0.05). The mortality rates of both of the twins, the heavier infant of the twins, and the lighter infant of the twins were lower in the 2013-2017 group compared with the 2008-2012 group (P<0.05). Compared with the 2008-2012 group, the 2013-2017 group (both the heavier infant and lighter infant subgroups) had lower incidence rates of pulmonary hemorrhage, patent ductus arteriosus (PDA), periventricular-intraventricular hemorrhage (P-IVH), and neonatal respiratory distress syndrome (NRDS) and a higher incidence rate of bronchopulmonary dysplasia (P<0.05). CONCLUSIONS: There is a significant increase in the survival rate over time in extremely preterm twins with a gestational age of <28 weeks in the 26 grade A tertiary hospitals in Guangdong Province. The incidences of severe asphyxia, pulmonary hemorrhage, PDA, P-IVH, and NRDS decrease in both the heavier and lighter infants of the twins, but the incidence of bronchopulmonary dysplasia increases. With the improvement of diagnosis and treatment, the multidisciplinary collaboration between different fields of fetal medicine including prenatal diagnosis, obstetrics, and neonatology is needed in the future to jointly develop management strategies for twin pregnancy.
Assuntos
Displasia Broncopulmonar , Síndrome do Desconforto Respiratório do Recém-Nascido , Displasia Broncopulmonar/epidemiologia , Feminino , Idade Gestacional , Humanos , Lactente , Lactente Extremamente Prematuro , Recém-Nascido , Gravidez , Síndrome do Desconforto Respiratório do Recém-Nascido/epidemiologia , Estudos Retrospectivos , Resultado do TratamentoRESUMO
Pregenomic RNA (pgRNA) is a direct transcription product of hepatitis B virus (HBV) covalently closed circular DNA (cccDNA), and it plays important roles in viral genome amplification and replication. This study was designed to investigate whether serum pgRNA is a strong alternative marker for reflecting HBV cccDNA levels and to analyze the correlation between serum pgRNA, serum HBV DNA, and hepatitis B surface antigen (HBsAg). A total of 400 HBV-infected patients who received nucleos(t)ide analog (NA) therapy with different clinical outcomes were involved in this research. Case groups included asymptomatic hepatitis B virus carrier (ASC), chronic hepatitis B (CHB), liver cirrhosis (LC), and hepatocellular carcinoma (HCC) patients, with 100 patients in each group. The results showed that the levels of HBV pgRNA had significant differences between these 4 groups. Serum pgRNA levels correlated well with serum HBV DNA and HBsAg levels (HBV pgRNA levels versus HBV DNA levels, r = 0.58, P < 0.001; HBV pgRNA levels versus HBsAg levels, r = 0.47, P < 0.001). In addition, we focused on the 108 HBV-infected patients with HBV DNA levels of <500 IU/ml; it was surprising to find that in 17.57% (13/74) of cases, HBV pgRNA could be detected even when the HBV DNA level was below 20 IU/ml. In conclusion, HBV pgRNA levels in serum can be a surrogate marker for intrahepatic HBV cccDNA compared with serum HBV DNA and HBsAg. The detection of serum HBV pgRNA levels may provide a reference for clinical monitoring of cccDNA levels and the selection of appropriate timing for discontinuing antiviral therapy, especially when HBV DNA levels are below the detection limit.
Assuntos
DNA Circular/sangue , Hepatite B/sangue , Hepatite B/diagnóstico , RNA Viral/sangue , Adulto , Idoso , Antivirais/uso terapêutico , Biomarcadores/sangue , Carcinoma Hepatocelular/tratamento farmacológico , Carcinoma Hepatocelular/virologia , Portador Sadio/diagnóstico , Portador Sadio/virologia , Feminino , Hepatite B/tratamento farmacológico , Antígenos de Superfície da Hepatite B/sangue , Antígenos de Superfície da Hepatite B/genética , Antígenos E da Hepatite B/sangue , Antígenos E da Hepatite B/genética , Vírus da Hepatite B/genética , Humanos , Fígado/virologia , Neoplasias Hepáticas/tratamento farmacológico , Neoplasias Hepáticas/virologia , Masculino , Pessoa de Meia-Idade , Carga Viral , Replicação Viral , Adulto JovemRESUMO
BACKGROUND: Alexander disease (AxD) is an autosomal-dominant leukodystrophy caused by heterozygous mutations in the glial fibrillary acidic protein (GFAP) gene. OBJECTIVES: The objective of this report is to characterize the clinical phenotype and identify the genetic mutation associated with adult-onset AxD. METHODS: A man presented with progressive unsteadiness since age 16. Magnetic resonance imaging findings revealed characteristic features of AxD. The GFAP gene was screened, and a candidate variant was functionally tested to evaluate causality. RESULTS: A homozygous c.197G > A (p.Arg66Gln) mutation was found in the proband, and his asymptomatic parents were heterozygous for the same mutation. This mutation affected GFAP solubility and promoted filament aggregation. The presence of the wild-type protein rescued mutational effects, consistent with the recessive nature of this mutation. CONCLUSIONS: This study is the first report of AxD caused by a homozygous mutation in GFAP. The clinical implication is while examining patients with characteristic features on suspicion of AxD, GFAP screening is recommended even without a supportive family history. © 2020 International Parkinson and Movement Disorder Society.
Assuntos
Doença de Alexander , Adolescente , Adulto , Doença de Alexander/diagnóstico por imagem , Doença de Alexander/genética , Proteína Glial Fibrilar Ácida/genética , Homozigoto , Humanos , Masculino , Mutação/genética , FenótipoRESUMO
Mutations in hepatitis B virus (HBV) reverse transcriptase (RT) are associated with nucleos(t)ide analogue (NA) resistance during long-term antiviral treatment. However, the characterization of mutations in HBV RT in untreated patients has not yet been well illustrated. The objective of this study was to investigate the characterization and clinical significance of natural variability in HBV RT in treatment-naive patients. HBV RT sequences were analyzed in 427 patients by Sanger sequencing and in 66 patients by next-generation sequencing. Primary or secondary NA resistance (NAr) mutations were not found, except A181T in RT (rtA181T) by Sanger sequencing, but they were detected by next-generation sequencing. Mutations were found in 56 RT amino acid (aa) sites by Sanger sequencing, 36 of which had mutations that could lead to changes in B or T cell epitopes in the RT or S protein. The distribution of mutations was diverse in different sections within the RT region. Multiple mutations showed significant association with HBV DNA, HBsAg, HBeAg, age, and severity of liver fibrosis. Mutations at rt251, rt266, rt274, rt280, rt283, rt284, and rt286 were found most in the advanced liver disease (ALD) group by next-generation sequencing. The present study demonstrates that next-generation sequencing (NGS) was more suitable than Sanger sequencing to monitor NAr mutations at a low rate in the treatment-naive patients, and that mutations in the RT region might be involved in the progression to ALD.
Assuntos
Variação Genética , Vírus da Hepatite B/genética , Hepatite B Crônica/virologia , Mutação , DNA Polimerase Dirigida por RNA/genética , Adulto , Idoso , Antivirais/uso terapêutico , Povo Asiático , China , DNA Viral/genética , Farmacorresistência Viral/genética , Feminino , Genótipo , Vírus da Hepatite B/enzimologia , Hepatite B Crônica/tratamento farmacológico , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Masculino , Pessoa de Meia-Idade , Análise de Sequência de DNA , Adulto JovemRESUMO
Timosaponin BII (TBII), a major steroidal saponin isolated from Anemarrhena asphodeloides Bge., displays a variety of promising pharmacological activities, such as neuroprotection, enhancement of learning and memory, vascular protection and inhibition of platelet aggregation; therefore, it has been developed as a pharmaceutical for prevention or treatment of dementia. Given the safety concerns surrounding timosaponins and the absence of studies on the safety of TBII, the potential toxicity of TBII was evaluated in toxicity and toxicokinetic studies in rats. In the acute oral toxicity study, loose stools were observed in rats receiving 4000 mg/kg, and the symptoms recovered within 1 day. In the 28-day repeated-dose oral toxicity and toxicokinetic study, rats receiving 540 mg/kg showed loose stools and a slight deceleration of body weight growth in both sexes, and the females also showed a slight decrease in food consumption. Moreover, urinalysis indicated reversible treatment-related toxicity in rats receiving 540 mg/kg. The toxicokinetic study demonstrated a dose-dependent increase in systematic exposure to TBII after 28 successive days of oral treatment with TBII. The accumulation coefficients of TBII were 4.35, 1.70 and 1.81, respectively, in rats that received 60, 180 and 540 mg/kg. The no-observed-adverse-effect level (NOAEL) is proposed to be 180 mg/kg.
Assuntos
Anemarrhena/química , Demência/tratamento farmacológico , Fármacos Neuroprotetores/farmacologia , Nível de Efeito Adverso não Observado , Saponinas/farmacologia , Esteroides/farmacologia , Administração Oral , Animais , Peso Corporal/efeitos dos fármacos , Relação Dose-Resposta a Droga , Feminino , Humanos , Masculino , Fármacos Neuroprotetores/uso terapêutico , Ratos , Ratos Sprague-Dawley , Saponinas/uso terapêutico , Esteroides/uso terapêutico , Testes de Toxicidade Aguda , Testes de Toxicidade Subcrônica , ToxicocinéticaRESUMO
BACKGROUND: Recent studies have highlighted the potential diagnostic values of microRNAs (miRNA) in various cancers involving oral cancer. This meta-analysis sought to summarize the global diagnostic accuracy of miRNAs for patients with oral cancer (OC). METHODS: A systematic review of multiple databases was performed to obtain original studies fulfilling search criteria and the quality of studies was assessed by the QUADAS tool. The bivariate meta-analysis model was employed to plot the summary receiver operator characteristic (SROC) curve. Influence analysis, meta-regression, and publication bias assay were all conducted using Stata 12.0 software. The trim-fill adjustment method was used to further assess the possible effect of publication bias. RESULTS: A total of 8 studies were included. The SROC analysis showed that miRNA profiling allowed for the discrimination between patients with high-risk oral lesions (OC or pre-cancer) and healthy donors, with a sensitivity of 0.84 (95% CI: 0.78-0.88) and specificity of 0.83 (95% CI: 0.78-0.87), corresponding to an area under curve (AUC) of 0.90. Our subgroup analyses suggested that miRNA signature harbored higher accuracy in diagnosing oral squamous cell carcinoma (OSCC) than pre-cancer lesions (AUC, sensitivity, and specificity of 0.90, 0.83, or 0.82, respectively). Moreover, stratified analyses revealed that parallel miRNA profiling, plasma- and Caucasian-based analyses all conferred promising accuracies for OC detection. The funnel plot assay manifested evidence of a publication bias. After the adjustment by the trim and fill method, the pooled adjusted efforts were slightly attenuated. CONCLUSIONS: MiRNA profiles hallmark a potential diagnostic value for detection of OC and potentially malignant disorders. Further studies should be performed to rigorously evaluate the diagnostic accuracy of miRNA profiling for OC.
Assuntos
MicroRNAs/análise , Neoplasias Bucais/diagnóstico , Biomarcadores Tumorais , Carcinoma de Células Escamosas/diagnóstico , Humanos , Neoplasias Bucais/genética , Viés de Publicação , Curva ROCRESUMO
The standard of 5-Hydroxymethylfurfural (5-HMF) existed in dextrose injection as an inevitable by-product during high-temperature setrilization has been included in pharmacopoeias considering its hazardous effects on human health. We found that the concentrations of 5-HMF in some traditional Chinese medicine injections (TCMIs) far exceeded its limit in dextrose injection. Besides, we detected 5, 5'-Oxydimethylenebis (2-furfural) (OMBF) in those TCMIs containing high concentrations of 5-HMF. We investigated the in vivo immunomodulatory effects of 5-HMF and OMBF at three dose levels using the reporter antigen popliteal lymph node assay (RA-PLNA), which allows the straightforward examination and mechanistic study of immunotoxicity of low molecular weight compounds. We found that 5-HMF increased the production of IgG2a and IFN-γ when co-injected with TNP-OVA, indicating its capability of providing a co-stimulatory signal to evoke a typical type-1 immune response. Compared with the 5-HMF, OMBF elevated the production of IgG1, IgG2, IL-4 and IFN-γ in response to both reporter antigens, suggesting that OMBF can act as a neo-antigen or neo-epitope to elicit a mixed type-1 and type-2 immune response. It indicates that both 5-HMF and OMBF have immunosensitizing potential with different mechanisms, and exposure to 5-HMF and OMBF may represent a safety concern for humans.
Assuntos
Furaldeído/análogos & derivados , Furaldeído/farmacologia , Fatores Imunológicos/farmacologia , Linfonodos/efeitos dos fármacos , Animais , Células Cultivadas , Relação Dose-Resposta a Droga , Furaldeído/química , Furaldeído/imunologia , Fatores Imunológicos/química , Fatores Imunológicos/imunologia , Ensaio Local de Linfonodo , Linfonodos/imunologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Estrutura Molecular , Relação Estrutura-AtividadeRESUMO
Sinoporphyrin sodium (DVDMS) is a novel hematoporphyrin-like photosensitizer developed for photodynamic therapy (PDT), an effective therapeutic modality for tumor treatment; however, the safety of photosensitizer-based PDT is always of great concern. The purpose of the current study was to investigate the potential repeated-dose toxicity and describe the toxicokinetic process of DVDMS-based PDT in Beagle dogs. The dogs were randomly allocated to six groups, and then were administrated a DVDMS preparation intravenously at dose levels of 0, 1, 3, 9, 1 and 9 mg per kg body weight, respectively; then, the latter two groups were illuminated 24 h later with a 630 nm laser for 10 min, once every seven days for 5 weeks. During the study period, clinical signs, mortality, body weight, food consumption, body temperature, ophthalmoscopy, hematology, serum biochemistry, urinalysis, electrocardiograms, toxicokinetics, organ weights, gross anatomy and histopathology were examined. After the administration, no deaths were observed; however, the dogs that received PDT showed skin swelling and ulceration, indicating that DVDMS-PDT induced a phototoxic effect. DVDMS led to an increase in blood coagulation in dogs in the 9 mg kg(-1) group and in the two PDT groups on Day 35, whereas it induced a decrease in dogs in the 3 mg kg(-1) group and in the two PDT groups on Day 49. The toxicokinetic study showed that the systematic exposure of DVDMS in dogs occurred in a dose-dependent manner, and DVDMS did not accumulate in blood plasma. The DVDMS-based PDT group showed no obvious treatment-related pathological changes; however, slight or mild brown-and-yellow pigmentation of DVDMS (or its metabolite) was observed to deposit in the liver, spleen, local lymph nodes and marrow of dogs in the mid- and high-dose groups, as well as the high-dose PDT group. In females, the absolute and relative spleen weights increased in dogs in the 9 mg kg(-1) DVDMS groups with and without PDT during the treatment and recovery period, respectively. The target organs are presumed to be the liver and immune organs (spleen, bone marrow and lymph nodes), while all of the responses were slight. Based on the results above, the no-observed-adverse-effect level (NOAEL) was considered to be 1 mg kg(-1), and DVDMS-PDT appeared to be a safe and promising anti-tumor therapy in the clinic.
Assuntos
Fármacos Fotossensibilizantes/toxicidade , Porfirinas/toxicidade , Animais , Coagulação Sanguínea/efeitos dos fármacos , Coagulação Sanguínea/efeitos da radiação , Temperatura Corporal/efeitos dos fármacos , Temperatura Corporal/efeitos da radiação , Peso Corporal/efeitos dos fármacos , Peso Corporal/efeitos da radiação , Cães , Relação Dose-Resposta a Droga , Ingestão de Alimentos/efeitos dos fármacos , Ingestão de Alimentos/efeitos da radiação , Feminino , Coração/efeitos dos fármacos , Coração/fisiologia , Coração/efeitos da radiação , Lasers , Masculino , Nível de Efeito Adverso não Observado , Fotoquimioterapia , Fármacos Fotossensibilizantes/administração & dosagem , Fármacos Fotossensibilizantes/química , Fármacos Fotossensibilizantes/farmacocinética , Porfirinas/administração & dosagem , Porfirinas/química , Porfirinas/farmacocinética , Distribuição Aleatória , Caracteres Sexuais , Dermatopatias/induzido quimicamente , Dermatopatias/etiologia , ToxicocinéticaRESUMO
OBJECTIVE: To discuss the role of Litchi chinensis seed saponins on hyperplasia of mammary glands(HMG) and the influence of estrogen signaling pathways in rats. METHODS: In addition to eight non-pregnant female SD rats in normal control group, the other 32 pathologic models of HMG rat model were randomly divided into other four groups: model control group, low and high dose groups, which were given experimental drug of Litchi chinensis seed saponins (LSE) 0. 1 g/kg and 0. 2 g/kg, and the positive control group with experimental drug of tamoxifen 4 mg/kg. Then all model rats were orally administrated for four weeks. The diameter and height of nipple were measured, and the content changes of estradiol(E2) and progestrogen(P) in serum were detected with ELISA method. The HMG were detected by the morphology examination. The expression of estrogen receptor(ER) and progesterone receptor(PR) in the mammary glands were investigated by immunohistochemical staining. RESULTS: According to LSE in the low and high dose groups, it was discovered that the nipple diameter and height of HMG rats, the expression of ER and PR in HMG and the content of serum E2 were reduced, the content of serum P were improved, and the hyperplasia of mammary glands was inhibited. CONCLUSION: LSE can obviously inhibit the rat hyperplasia of mammary gland, and its possible mechanism is related to adjusting the transduction pathway of estrogen signal to lower estrogen levels.
Assuntos
Medicamentos de Ervas Chinesas/farmacologia , Estrogênios/metabolismo , Litchi/química , Glândulas Mamárias Animais/efeitos dos fármacos , Saponinas/farmacologia , Transdução de Sinais/efeitos dos fármacos , Animais , Estradiol/sangue , Feminino , Hiperplasia , Glândulas Mamárias Animais/patologia , Progesterona/sangue , Ratos , Ratos Sprague-Dawley , Sementes/químicaRESUMO
AIMS: The aim of this study was to evaluate the prevalence of PBK/TOPK (PDZ-binding kinase/T-LAK cell-originated protein kinase) expression, and explore the prognostic significance of PBK/TOPK expression alone and in combination with Ki67 and p53 expression in non-small-cell lung cancer (NSCLC). METHODS AND RESULTS: We detected PBK/TOPK expression in 30 samples of normal lung tissue, 32 lymph node metastases and 279 primary non-small-cell lung cancers by immunohistochemistry, and analysed the correlation of PBK/TOPK expression with Ki67 and p53 expression in primary tumour tissues. The results showed that PBK/TOPK expression was higher in lymph node metastases (75%) than in primary tumours (44.8%) and normal lung tissues (0%). PBK/TOPK expression was associated with histological type, lymph node metastasis, and TNM stage, and was positively correlated with Ki67 and p53 expression in NSCLC. Univariate and multivariate survival analyses showed that PBK/TOPK expression was significantly associated with an unfavourable prognosis in NSCLC. The prognosis of patients with tumours positive for both PBK/TOPK expression and Ki67 or p53 expression was also significantly unfavourable. CONCLUSIONS: PBK/TOPK expression is positively correlated with Ki67 and p53 expression, and can be used as an independent prognostic factor in NSCLC.
Assuntos
Carcinoma Pulmonar de Células não Pequenas/metabolismo , Neoplasias Pulmonares/metabolismo , Pulmão/metabolismo , Quinases de Proteína Quinase Ativadas por Mitógeno/metabolismo , Adulto , Idoso , Biomarcadores Tumorais/metabolismo , Carcinoma Pulmonar de Células não Pequenas/patologia , Feminino , Humanos , Antígeno Ki-67/metabolismo , Pulmão/patologia , Neoplasias Pulmonares/patologia , Masculino , Pessoa de Meia-Idade , Estadiamento de Neoplasias , Prognóstico , Proteína Supressora de Tumor p53/metabolismoRESUMO
HAb18G, a novel cancer biomarker, has been shown to be involved in the progression of malignancy by regulating expression of vascular endothelial growth factor (VEGF) and matrixmetalloproteinases (MMPs). The goal of this study was to evaluate the role of HAb18G in the biology of NSCLC and to determine its potential as a therapeutic target. HAb18G protein expression was detected by immunohistochemistry in 150 NSCLC tissues. The results showed that HAb18G protein expression was associated with tumor diameter, lymph node status, tumor stage, and poor prognosis (P < 0.05). Multivariate analysis showed that HAb18G overexpression was an independent prognostic factor (HR, 3.713; 95 % CI, 1.114-12.373; P = 0.033). Transient infection of A549 lung cancer cells with small interfering RNA (SiRNA) against HAb18G efficiently inhibited the expression of HAb18G in A549 lung cancer cells at both mRNA and protein levels. Downregulation of HAb18G not only reduced MMP-2, MMP-9, and VEGF at mRNA and protein levels in A549 cells, but also inhibited fibroblasts to secrete MMP-2 and MMP-9 at mRNA level. Additionally, downregulation of HAb18G mRNA resulted in decreased migration, proliferation, and increased apoptosis of A549 in vitro. Our findings suggest that HAb18G overexpression plays an important role in progression of NSCLC and HAb18G may be a potential target of NSCLC therapy.
Assuntos
Apoptose , Basigina/metabolismo , Carcinoma Pulmonar de Células não Pequenas/patologia , Movimento Celular , Regulação para Baixo , Neoplasias Pulmonares/patologia , RNA Interferente Pequeno/metabolismo , Adulto , Idoso , Idoso de 80 Anos ou mais , Apoptose/genética , Basigina/genética , Carcinoma Pulmonar de Células não Pequenas/enzimologia , Carcinoma Pulmonar de Células não Pequenas/genética , Linhagem Celular Tumoral , Movimento Celular/genética , Proliferação de Células , Técnicas de Cocultura , Feminino , Fibroblastos/enzimologia , Fibroblastos/patologia , Regulação Neoplásica da Expressão Gênica , Humanos , Estimativa de Kaplan-Meier , Neoplasias Pulmonares/enzimologia , Neoplasias Pulmonares/genética , Masculino , Metaloproteinase 2 da Matriz/genética , Metaloproteinase 2 da Matriz/metabolismo , Metaloproteinase 9 da Matriz/genética , Metaloproteinase 9 da Matriz/metabolismo , Pessoa de Meia-Idade , Modelos de Riscos Proporcionais , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Fator A de Crescimento do Endotélio Vascular/genética , Fator A de Crescimento do Endotélio Vascular/metabolismo , CicatrizaçãoRESUMO
Chronic hepatitis B (CHB) patients treated with interferon shows encouraging results. However, its clinical efficacy is limited by significant individual differences in treatment responses. We identified an interferon-inducible effector, TRIM22, as the likely causal target of such differential responses. We found that TRIM22 was highly expressed in interferon-responsive patients and negatively correlated with HBV DNA and HBeAg serum levels. Stable cells overexpressing TRIM22 carried significantly less HBsAg, HBeAg, and HBV DNA, and cells with knocked-down TRIM22 by shRNA displayed higher levels of these markers than controls. Integrated bioinformatics analysis and subsequent experiments revealed that TRIM22 overexpression significantly increased the supernatant levels of IL-1ß and IL-8, two important cytokines of NOD2/NF-κB pathway involved in interferon-induced antiviral activities. We identified three candidate microRNAs binding to 3'UTR of TRIM22 at various locations through typical imperfect paring using the TargetScan program. MiR-548c-3p appeared to be highly expressed, while the TRIM22 level was low in the suboptimal response group of CHB patients. The Luciferase reporter assay revealed an interaction between miR-548c-3p and the 3'UTR of TRIM22, leading to a controlled suppression of TRIM22 endogenous expression. This resulted in interferon's substantially weakened therapeutic efficacy, as indicated by the elevation of the serum levels of HBsAg, HBeAg and HBV DNA in miR-548c-3p-transfected HepAD38 cells. Our study demonstrated that a particular miR-548c-3p is the key negative regulator of TRIM22 in CHB patients with a weak response to interferon treatment, providing a novel marker and target in interferon-α therapy evaluation.
Assuntos
Hepatite B Crônica , MicroRNAs , Humanos , Antígenos E da Hepatite B , Antígenos de Superfície da Hepatite B , Hepatite B Crônica/tratamento farmacológico , DNA Viral , Regiões 3' não Traduzidas , Interferon-alfa/uso terapêutico , Antivirais/farmacologia , Antivirais/uso terapêutico , Resultado do Tratamento , MicroRNAs/genética , Polietilenoglicóis/uso terapêutico , Proteínas Recombinantes/uso terapêutico , Proteínas com Motivo Tripartido/genética , Proteínas Repressoras , Antígenos de Histocompatibilidade Menor/genéticaRESUMO
Background: Alzheimer's disease (AD) is a multifactorial disorder characterized by cognitive decline. Current available therapeutics for AD have limited clinical benefit. Therefore, preventive therapies for interrupting the development of AD are critically needed. Molecules targeting multifunction to interact with various pathlogical components have been considered to improve the therapeutic efficiency of AD. In particular, herbal medicines with multiplicity of actions produce cognitive benefits on AD. Bugu-M is a multi-herbal extract composed of Ganoderma lucidum (Antler form), Nelumbo nucifera Gaertn., Ziziphus jujuba Mill., and Dimocarpus longan, with the ability of its various components to confer resilience to cognitive deficits. Objective: To evaluate the potential of Bugu-M on amyloid-ß (Aß) toxicity and its in vitro mechanisms and on in vivo cognitive function. Methods: We illustrated the effect of Bugu-M on Aß25-35-evoked toxicity as well as its possible mechanisms to diminish the pathogenesis of AD in rat cortical neurons. For cognitive function studies, 2-month-old female 3×Tg-AD mice were administered 400âmg/kg Bugu-M for 30 days. Behavioral tests were performed to assess the efficacy of Bugu-M on cognitive impairment. Results: In primary cortical neuronal cultures, Bugu-M mitigated Aß-evoked toxicity by reducing cytoskeletal aberrations and axonal disruption, restoring presynaptic and postsynaptic protein expression, suppressing mitochondrial damage and apoptotic signaling, and reserving neurogenic and neurotrophic factors. Importantly, 30-day administration of Bugu-M effectively prevented development of cognitive impairment in 3-month-old female 3×Tg-AD mice. Conclusion: Bugu-M might be beneficial in delaying the progression of AD, and thus warrants consideration for its preventive potential for AD.
RESUMO
IMPORTANCE: Pegylated interferon alfa (PegIFNα) has limited efficacy in the treatment of chronic hepatitis B (CHB). Although many biomarkers related to hepatitis B virus (HBV) have been proposed to stratify patients, the response rate to PegIFNα is still unsatisfactory. Herein, our data suggest that the single-nucleotide polymorphism (SNP) rs10838543 in TRIM22 potentiates a positive clinical response to PegIFNα treatment in patients with hepatitis B e antigen-positive CHB by increasing the levels of IFNL1, CCL3, and CCL5. These observations can help guide treatment decisions for patients with CHB to improve the response rate to PegIFNα.