RESUMO
BACKGROUND: Identifying valid biomarkers for patient selection impressively promotes the success of anti-PD-1 therapy. However, the unmet need for biomarkers in gastrointestinal (GI) cancers remains significant. We aimed to explore the predictive value of the circulating T-cell receptor (TCR) repertoire for clinical outcomes in GI cancers who received anti-PD-1 therapy. METHODS: 137 pre- and 79 post-treated peripheral blood samples were included. The TCR repertoire was evaluated by sequencing of complementarity-determining region 3 (CDR3) in the TRB gene. The Shannon index was used to measure the diversity of the TCR repertoire, and Morisita's overlap index was used to determine TCR repertoire similarities between pre- and post-treated samples. RESULTS: Among all enrolled patients, 76 received anti-PD-1 monotherapy and 61 received anti-PD-1 combination therapy. In the anti-PD-1 monotherapy cohort, patients with higher baseline TCR diversity exhibited a significantly higher disease control rate (77.8% vs. 47.2%; hazard ratio [HR] 3.92; 95% confidence interval [CI] 1.14-13.48; P = 0.030) and a longer progression-free survival (PFS) (median: 6.47 months vs. 2.77 months; HR 2.10; 95% CI 1.16-3.79; P = 0.014) and overall survival (OS) (median: NA vs. 8.97 months; HR 3.53; 95% CI 1.49-8.38; P = 0.004) than those with lower diversity. Moreover, patients with a higher TCR repertoire similarity still showed a superior PFS (4.43 months vs. 1.84 months; HR 13.98; 95% CI 4.37-44.68; P < 0.001) and OS (13.40 months vs. 6.12 months; HR 2.93; 95% CI 1.22-7.03; P = 0.016) even in the cohort with lower baseline diversity. However, neither biomarker showed predictive value in the anti-PD-1 combination therapy cohort. Interestingly, the combination of TCR diversity and PD-L1 expression can facilitate patient stratification in a pooled cohort. CONCLUSION: The circulating TCR repertoire can serve as a predictor of clinical outcomes in anti-PD-1 therapy in GI cancers.
Assuntos
Antineoplásicos Imunológicos/uso terapêutico , Neoplasias Gastrointestinais/terapia , Inibidores de Checkpoint Imunológico/uso terapêutico , Imunoterapia Adotiva/métodos , Receptores de Antígenos de Linfócitos T/sangue , Adulto , Idoso , Antígeno B7-H1/metabolismo , Biomarcadores Tumorais/sangue , Regiões Determinantes de Complementaridade/genética , Feminino , Neoplasias Gastrointestinais/sangue , Neoplasias Gastrointestinais/mortalidade , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Masculino , Pessoa de Meia-Idade , Intervalo Livre de Progressão , Receptores de Antígenos de Linfócitos T alfa-beta/sangue , Fatores de Tempo , Resultado do Tratamento , Adulto JovemRESUMO
In this study, neem leaves were successively extracted with petroleum ether, 95% ethanol and water and the insecticidal activities of these extracts against Oxya chinensis larvae were measured. The results showed that 95% ethanol extract gave the highest extraction yield and insecticidal activity, and it was further extracted with five different solvents. The petroleum ether extract from the 95% ethanol extract possessed the highest insecticidal activity with median lethal concentration values ranging from 14.93 to 55.66mg/mL. The gas chromatography-mass spectrometer analysis showed that the petroleum ether extract mainly composed of alkanes, olefin, esters and amide. The pathological examination revealed that the prominent lesions, including reduced regenerative cells in midgut and swelled and degenerated cylindrical cells, were observed in the 5th instar Oxya chinensis after treatment. The ultrastructural features showed that the cylindrical cells, microvilli and mitochondria were seriously damaged. These results suggested that the petroleum ether extract from neem leaves had potent insecticidal activity and could be a candidate insecticide.(AU)
Nesse estudo, folhas "neem" foram extraídas com sucesso com éter de petróleo, 95% de etanol e água, e as atividades inseticidas desses extratos foram medidas contra larvas de Oxya chinesis. Os resultados mostram que extrato com 95% de etanol deram o maior resultado de extração e atividade inseticida e foi então extraído utilizando mais cinco diferentes solventes. O éter de petróleo do extrato de 95% etanol apresentou maior atividade inseticida com concentração letal média variando de 14.93 a 55.66mg/mL. A análise por cromatografia de massa mostrou que o extrato de éter de petróleo está composto principalmente de alcanos, alcenos, ésteres e amidas. A avaliação patológica revelou que as lesões proeminentes, inclusive células regenerativas reduzidas no intestino e células cilíndricas edemaciadas e degeneradas foram observadas no quinto estágio de desenvolvimento da Oxya chinesis após tratamento. As características ultraestruturais mostraram que as células cilíndricas, microvilos e mitocôndrias apresentavam lesões graves. Esses resultados sugerem que o extrato de éter de petróleo de folhas de "neem" tem atividade inseticida potente e pode ser um candidato a inseticida.(AU)
Assuntos
Azadirachta/fisiologia , Inseticidas/análiseRESUMO
In this study, neem leaves were successively extracted with petroleum ether, 95% ethanol and water and the insecticidal activities of these extracts against Oxya chinensis larvae were measured. The results showed that 95% ethanol extract gave the highest extraction yield and insecticidal activity, and it was further extracted with five different solvents. The petroleum ether extract from the 95% ethanol extract possessed the highest insecticidal activity with median lethal concentration values ranging from 14.93 to 55.66mg/mL. The gas chromatography-mass spectrometer analysis showed that the petroleum ether extract mainly composed of alkanes, olefin, esters and amide. The pathological examination revealed that the prominent lesions, including reduced regenerative cells in midgut and swelled and degenerated cylindrical cells, were observed in the 5th instar Oxya chinensis after treatment. The ultrastructural features showed that the cylindrical cells, microvilli and mitochondria were seriously damaged. These results suggested that the petroleum ether extract from neem leaves had potent insecticidal activity and could be a candidate insecticide.(AU)
Nesse estudo, folhas "neem" foram extraídas com sucesso com éter de petróleo, 95% de etanol e água, e as atividades inseticidas desses extratos foram medidas contra larvas de Oxya chinesis. Os resultados mostram que extrato com 95% de etanol deram o maior resultado de extração e atividade inseticida e foi então extraído utilizando mais cinco diferentes solventes. O éter de petróleo do extrato de 95% etanol apresentou maior atividade inseticida com concentração letal média variando de 14.93 a 55.66mg/mL. A análise por cromatografia de massa mostrou que o extrato de éter de petróleo está composto principalmente de alcanos, alcenos, ésteres e amidas. A avaliação patológica revelou que as lesões proeminentes, inclusive células regenerativas reduzidas no intestino e células cilíndricas edemaciadas e degeneradas foram observadas no quinto estágio de desenvolvimento da Oxya chinesis após tratamento. As características ultraestruturais mostraram que as células cilíndricas, microvilos e mitocôndrias apresentavam lesões graves. Esses resultados sugerem que o extrato de éter de petróleo de folhas de "neem" tem atividade inseticida potente e pode ser um candidato a inseticida.(AU)
Assuntos
Azadirachta/fisiologia , Inseticidas/análiseRESUMO
To study the development rules of Chinese native geese, two breeds, Shitou and Sichuan White geese were analyzed from 0 to 12 weeks of age. The growth curves were fitted with commonly used four kinds of nonlinear models (Logistic, Gompertz, Von Bertalanffy and Richards). The results showed that the growth curves were appropriately fitted with all four models but the Logistic and Richards both had the best fitting with growth curve (R2>0.99). Analyzing the fitting parameters of the Logistic and Richards, we found that male Shitou had the highest adult body weight while Sichuan White female had the lowest weight. In Shitou breed, Shape parameter Predicted with Richards model was corresponded with Gompertz curve, while in Sichuan breed it was in between Gompertz and Bertalanffy. Growth parameters predicted with Logistic model was much more closed to observed value as compared others. So overall logistic was the best model to analyze the growth curve in Chinese native goose and Shitou goose had excellent growth performance when compared to Sichuan White.(AU)
Assuntos
Animais , Recém-Nascido , Gansos/crescimento & desenvolvimento , Gansos/fisiologia , Crescimento/fisiologia , Peso FetalRESUMO
To study the development rules of Chinese native geese, two breeds, Shitou and Sichuan White geese were analyzed from 0 to 12 weeks of age. The growth curves were fitted with commonly used four kinds of nonlinear models (Logistic, Gompertz, Von Bertalanffy and Richards). The results showed that the growth curves were appropriately fitted with all four models but the Logistic and Richards both had the best fitting with growth curve (R2>0.99). Analyzing the fitting parameters of the Logistic and Richards, we found that male Shitou had the highest adult body weight while Sichuan White female had the lowest weight. In Shitou breed, Shape parameter Predicted with Richards model was corresponded with Gompertz curve, while in Sichuan breed it was in between Gompertz and Bertalanffy. Growth parameters predicted with Logistic model was much more closed to observed value as compared others. So overall logistic was the best model to analyze the growth curve in Chinese native goose and Shitou goose had excellent growth performance when compared to Sichuan White.
Assuntos
Animais , Recém-Nascido , Crescimento/fisiologia , Gansos/crescimento & desenvolvimento , Gansos/fisiologia , Peso FetalRESUMO
Genomic imprinting is an important epigenetic mechanism that has vital effects on fetal growth and development. We observed the differences in four tissues (heart, spleen, liver, and kidney) from dead transgenic cloned goats using hematoxylin and eosin (H&E) staining. Eight imprinted genes in the tissues of dead transgenic cloned and normal goats were analyzed using reverse transcription polymerase chain reaction. H&E staining results from the abortion group indicated the lack of obvious morphological changes in heart and spleen tissues, while inflammatory cell infiltration and glomerular nephritis characteristics were observed in liver and kidney tissues, respectively. Compared to the control group, CDKN1C, H19, IGF2R, and SNRPN were significantly (P < 0.05) overexpressed in the heart tissue of the abortion group, while XIST was significantly reduced. In the liver tissues, CDKN1C and DLK1 expression decreased, while GNAS, H19, IGF2R, PEG3, and XIST expression increased significantly. In the spleen tissues, DLK1 expression increased, while GNAS, H19, IGF2R, PEG3, SNRPN, and XIST expression decreased. In the kidney tissues, CDKN1C, DLK1, GNAS, IGF2R, and PEG3 expression increased, while H19 and XIST expression decreased. The overall expression of imprinted genes was abnormal in different tissues of transgenic cloned goats, and the degree of abnormal genomic imprinting was more severe in the abortion group compared to the death and control groups. These results suggest that abnormal expression of imprinted genes may cause developmental defects in transgenic cloned goats. Moreover, abnormal epigenetic modifications may affect the reprogramming of transgenic donor cells.
Assuntos
Clonagem de Organismos/mortalidade , Epigênese Genética , Genes Letais , Impressão Genômica , Cabras/genética , Lactoferrina/genética , Animais , Animais Geneticamente Modificados , Inibidor de Quinase Dependente de Ciclina p57/genética , Inibidor de Quinase Dependente de Ciclina p57/metabolismo , Feminino , Perfilação da Expressão Gênica , Cabras/metabolismo , Humanos , Peptídeos e Proteínas de Sinalização Intercelular/genética , Peptídeos e Proteínas de Sinalização Intercelular/metabolismo , Rim/metabolismo , Fatores de Transcrição Kruppel-Like/genética , Fatores de Transcrição Kruppel-Like/metabolismo , Lactoferrina/metabolismo , Fígado/metabolismo , Masculino , Miocárdio/metabolismo , Proteínas do Tecido Nervoso/genética , Proteínas do Tecido Nervoso/metabolismo , Gravidez , Transdução de Sinais , Baço/metabolismo , TransgenesRESUMO
PURPOSE: This study seeks to evaluate the natural history, outcome, and possible prognostic factors in patients with brain metastases derived from gastrointestinal cancers. METHODS: The clinical features, prognostic factors, and the effects of different treatment modalities on survival were retrospectively investigated in 103 patients with brain metastases derived from gastrointestinal cancers. RESULTS: The median time from diagnosis of primary tumor to brain metastasis was 22.00 months. The interval between diagnosis of primary tumor relapse and brain metastasis was 8.00 months. The median follow-up time was 7.80 months. The median survival time after diagnosis of brain metastases was 4.10 months for all patients and 1.17 months for patients who received only steroids (36.9 %), 3.97 months for patients who only received whole-brain radiation therapy (WBRT 31.1 %), 11.07 months for patients who received gamma-knife surgery alone or/and WBRT (20.4 %), and 13.70 months for patients who underwent surgery and radiotherapy (12 patients, 11.6 %) (P < 0.001). Multivariate analysis revealed that recursive partitioning analysis (RPA) class, extracranial metastasis, and chemotherapy were independent prognostic factors. Brain metastasis derived from gastrointestinal tract cancer is rare, and overall patient survival is poor. CONCLUSION: RPA class, chemotherapy after brain metastases, and treatment regimens were independent prognostic factors for the survival of patients with brain metastases derived from gastrointestinal cancers.
Assuntos
Neoplasias Encefálicas/diagnóstico , Neoplasias Encefálicas/secundário , Neoplasias Gastrointestinais/diagnóstico , Neoplasias Gastrointestinais/patologia , Adulto , Idoso , Neoplasias Encefálicas/mortalidade , Neoplasias Encefálicas/terapia , Irradiação Craniana , Feminino , Neoplasias Gastrointestinais/mortalidade , Neoplasias Gastrointestinais/terapia , Humanos , Masculino , Pessoa de Meia-Idade , Prognóstico , Radiocirurgia , Estudos Retrospectivos , Análise de Sobrevida , Resultado do TratamentoRESUMO
The US2 protein has been reported to contribute to duck enteritis virus (DEV) infection; however, its kinetics and localization during infection, and whether it is a component of virion, have not been previously reported. To elucidate the function of DEV US2, US2 was amplified by polymerase chain reaction (PCR) and inserted into pET-32a(+); this was expressed, the recombinant US2 protein was purified, and a polyclonal antibody generated. In addition, the kinetics and localization of the US2 gene and protein were determined by quantitative real-time fluorescent PCR, ganciclovir (GCV), and cycloheximide (CHX) treatment, western-blot, and indirect immunofluorescence assay. The packaging of US2 into DEV virions was revealed by a protease protection assay. US2 was found to be transcribed 24 h post-infection (pi) and peaked at 72 h pi; the US2 protein was detected 48 h pi, except in the presence of GCV or CHX. US2 was packed into virions and also localized to the plasma membrane and cytoplasm in infected cells. The results showed that the DEV US2 is a late gene, and that its encoding protein could be a tegument component localized mainly in the cytoplasm. This study provides useful data for the further analysis of DEV US2, including an explanation for the genetic conservation among alphaherpesviruses.
Assuntos
Mardivirus/genética , Proteínas do Envelope Viral/genética , Animais , Linhagem Celular , Patos , Fibroblastos , Expressão Gênica , Mardivirus/efeitos dos fármacos , Transporte Proteico , Proteínas Recombinantes , Proteínas do Envelope Viral/isolamento & purificação , Proteínas do Envelope Viral/metabolismo , VírionRESUMO
Dairy goat is a good model for production of transgenic proteins in milk using somatic cell nuclear transfer (SCNT). However, animals produced from SCNT are often associated with lung deficiencies. We recently produced six transgenic cloned dairy goats harboring the human lactoferrin gene, including three live transgenic clones and three deceased transgenic clones that died from respiratory failure during the perinatal period. Imprinted genes are important regulators of lung growth, and may be subjected to faulty reprogramming. In the present study, first, microsatellite analysis, PCR, and DNA sequence identification were conducted to confirm that these three dead kids were genetically identical to the transgenic donor cells. Second, the CpG island methylation profile of the imprinted insulin-like growth factor receptor (IGF2R) gene was assessed in the lungs of the three dead transgenic kids and the normally produced kids using bisulfite sequencing PCR. In addition, the relative mRNA level of IGF2R was also determined by real-time PCR. Results showed that the IGF2R gene in the lungs of the dead cloned kids showed abnormal hypermethylation and higher mRNA expression levels than the control, indicating that aberrant DNA methylation reprogramming is one of the important factors in the death of transgenic cloned animals.
Assuntos
Cabras/genética , Lactoferrina/genética , Pulmão/metabolismo , Receptor IGF Tipo 2/genética , Animais , Animais Geneticamente Modificados , Sequência de Bases , Clonagem de Organismos , Metilação de DNA , Transferência Embrionária , Feminino , Expressão Gênica , Impressão Genômica , Humanos , Repetições de Microssatélites , Dados de Sequência Molecular , Receptor IGF Tipo 2/metabolismo , Análise de Sequência de DNARESUMO
Numerous studies have evaluated the association between Arg389Gly polymorphism in the ß1 adrenergic receptor gene and heart failure risk. However, the specific association is still controversial. We performed a meta-analysis of all case-control studies that evaluated the association between Arg389Gly polymorphism and heart failure in humans. Studies were identified in the PubMed, Embase, and China National Knowledge Infrastructure databases. Two reviewers independently assessed the studies. Six case-control studies with a total of 1736 participants were included in the meta-analysis, including 882 cases with heart failure and 854 controls, and our results showed no association between the Arg389Gly polymorphism and heart failure [ArgArg vs GlyGly: odds ratio (OR) = 0.84, 95% confidence interval (CI) 0.59-1.20; ArgArg vs ArgGly: OR = 0.95, 95%CI 0.78-1.16; dominant model: OR = 1.08, 95%CI 0.89-1.31; recessive model: OR = 0.96, 95%CI 0.69-1.35]. No publication bias was found in the present study (all P values > 0.05). In conclusion, the ß1 adrenergic receptor gene Arg389Gly polymorphism might not be associated with heart failure risk. Further large and well-designed studies are needed to confirm this conclusion.
Assuntos
Predisposição Genética para Doença , Insuficiência Cardíaca/genética , Hipertensão/genética , Receptores Adrenérgicos beta 1/genética , China , Feminino , Estudos de Associação Genética , Insuficiência Cardíaca/patologia , Humanos , Hipertensão/patologia , Polimorfismo de Nucleotídeo Único , Fatores de RiscoRESUMO
Several published articles have evaluated the association between the prostate stem cell antigen (PSCA) rs2294008 (C>T) polymorphism and bladder cancer risk, but the results remain inconclusive. In order to derive a more precise estimation of the association, we performed a meta-analysis of four case-control studies that included 9617 cases and 16,323 controls. Odds ratios (ORs) and 95% confidence intervals (CIs) were used to assess the strength of the association. Our meta-analysis showed that, overall, the rs2294008 (C>T) polymorphism was associated with bladder cancer susceptibility (OR = 1.29, 95%CI = 1.20-1.40 for TT vs CC; OR = 1.24, 95%CI = 1.16-1.31 for CT vs CC; OR = 1.25, 95%CI = 1.18-1.33 for TT/CT vs CC; OR = 1.13, 95%CI = 1.06-1.20 for TT vs CT/CC). In the stratified analyses, the risk remained significant for studies of European populations, Asian populations, population-based studies, and hospital-based studies. In conclusion, the results suggest that the PSCA rs2294008 (C>T) polymorphism is a risk factor for bladder cancer development.
Assuntos
Antígenos de Neoplasias/genética , Predisposição Genética para Doença , Proteínas de Neoplasias/genética , Polimorfismo de Nucleotídeo Único , Neoplasias da Bexiga Urinária/genética , Alelos , Estudos de Casos e Controles , Proteínas Ligadas por GPI/genética , Genótipo , Humanos , Masculino , Razão de Chances , Viés de Publicação , RiscoRESUMO
Mesenchymal stem cells derived from bone marrow (BMSCs) are a population of self-renewing multipotent cells that are capable of differentiating into various cellular lineages, and are widely employed in tissue engineering and cell therapy. Recently, clinical research involving BMSCs has become increasingly popular. In order to conduct appropriate research, it is first necessary to amplify large amounts of functional BMSCs in vitro. However, after several passages of expanding in vitro, the proliferation and differentiation potential of BMSCs gradually decline. To determine whether overexpression of Oct4 or Sox2 might prevent this decline, we transfected Oct4 or Sox2, which are essential for the pluripotency and self-renewal of embryonic stem cells, into BMSCs of Xiaomeishan porcine by a lentivirus. The results showed that overexpression of Sox2 or Oct4 BMSCs in culture media containing a basic fibroblast growth factor resulted in higher proliferation and differentiation compared to controls, suggesting that genetic modification of stemness-related genes is an efficient way to maintain the proliferation and differentiation potential of BMSCs.
Assuntos
Adipogenia , Proliferação de Células , Células-Tronco Mesenquimais/fisiologia , Fator 3 de Transcrição de Octâmero/metabolismo , Fatores de Transcrição SOXB1/metabolismo , Fosfatase Alcalina/metabolismo , Animais , Células Cultivadas , Fatores de Crescimento de Fibroblastos/fisiologia , Expressão Gênica , Células HEK293 , Humanos , Fator 3 de Transcrição de Octâmero/genética , Osteogênese , Fatores de Transcrição SOXB1/genética , Sus scrofaRESUMO
The Shanyi inbred A and E strains of the Chinese hamster are widely used in biomedical research, but detailed genetic characterization has been lacking. We developed microsatellite markers that could be used for genetic diversity analysis and linkage map construction. We isolated and characterized 16 novel microsatellite loci from a microsatellite-enriched genomic DNA library. These loci were genotyped in 48 animals from the two strains, and the polymorphic information content was determined. In the Shanyi A and E populations, 14 and 15 loci were found to be polymorphic, respectively, with polymorphic information content ranging from 0.1393 to 0.8082 and from 0.1109 to 0.7397, respectively. A total of 115 alleles were found for the 16 microsatellite loci in the two populations; the mean observed heterozygosity (H(O)) was 0.5191 and 0.4333 for the A and E populations, respectively, indicating marked genetic variation within the two populations. Correspondingly, the F(ST) values ranged from 0.002 to 0.9253, with an overall mean of 0.1935, indicating significant genetic difference between the two strains. The population differentiation levels were substantiated by Nei's genetic distance and full Bayesian analyses computed with STRUCTURE. Despite the genetic diversity and differentiation within and between the two inbred populations, the 48 individuals were correctly allocated into their original populations with high statistical confidence based on these 16 microsatellite loci. These novel microsatellite loci should be useful genetic markers for these two Chinese hamster inbred strains.