RESUMEN
Brucellosis has become a global zoonotic disease, seriously endangering the health of people all over the world. Vaccination is an effective strategy for protection against Brucella infection in livestock in developed countries. However, current vaccines are pathogenic to humans and pregnant animals, which limits their use. Therefore, it is very important to improve the safety and immune protection of Brucella vaccine. In this study, different bioinformatics approaches were carried out to predict the physicochemical properties, T/B epitope, and tertiary structure of Omp2b and Omp31. Then, these two proteins were sequentially linked, and the Cytotoxic T lymphocyte associated antigen-4 (CTLA-4) variable region was fused to the N-terminal of the epitope sequence. In addition, molecular docking was performed to show that the structure of the fusion protein vaccine had strong affinity with B7 (B7-1, B7-2). This study showed that the designed vaccine containing CTLA-4 had high potency against Brucella, which could provide a reference for the future development of efficient brucellosis vaccines.
Asunto(s)
Vacunas Bacterianas , Brucelosis , Antígeno CTLA-4 , Brucelosis/prevención & control , Brucella , Vacunas Bacterianas/inmunología , Antígeno CTLA-4/inmunología , Humanos , Animales , Epítopos/inmunología , Simulación del Acoplamiento Molecular , Biología Computacional , Proteínas Bacterianas/inmunología , Secuencia de Aminoácidos , Estructura Terciaria de Proteína , Proteínas Recombinantes de Fusión/inmunologíaRESUMEN
Brucellosis has become a global zoonotic disease, seriously endangering the health of people all over the world. Vaccination is an effective strategy for protection against Brucella infection in livestock in developed countries. However, current vaccines are pathogenic to humans and pregnant animals, which limits their use. Therefore, it is very important to improve the safety and immune protection of Brucella vaccine. In this study, different bioinformatics approaches were carried out to predict the physicochemical properties, T/B epitope, and tertiary structure of Omp2b and Omp31. Then, these two proteins were sequentially linked, and the Cytotoxic T lymphocyte associated antigen-4 (CTLA-4) variable region was fused to the N-terminal of the epitope sequence. In addition, molecular docking was performed to show that the structure of the fusion protein vaccine had strong affinity with B7 (B7-1, B7-2). This study showed that the designed vaccine containing CTLA-4 had high potency against Brucella, which could provide a reference for the future development of efficient brucellosis vaccines.
RESUMEN
The objective of the present study was to investigate the different plasma metabolites between anestrus and estrus postpartum dairy cows and to provide a theoretical basis for prevention of anestrus in dairy farm cows. In the experiment, one hundred and sixty-seven Holstein dairy cows were selected with similar age and parity. According to the concentration of ß-hydroxybutyric acid, non-esterified fatty acids and glucose in plasma during 14 to 21 days in milk, all dairy cows were determined as having a status of energy balance. According to the results of clinical symptom, rectal and B ultrasound examination at 60 to 90 days postpartum, these cows were divided into twenty estrus and twenty-four anestrus group, other dairy cows were removed. 1H nuclear magnetic resonance technology was utilized to detect the plasma metabolites changes and screen different plasma metabolites between anestrus and estrus cows. Ten different metabolites including alanine, glutamic acid, asparagine, creatine, choline, phosphocholine, glycerophosphocholine, low-density lipoprotein, and very-low-density lipoprotein were significantly decreased in anestrous cows compared with estrous cows. Metabolic pathway analyses indicated that differential metabolites were primarily involved in amino acid and glycerophospholipid metabolism. These metabolites and their enrichment pathways indicate that reduced steroid hormone synthesis precursors result in lower levels of estradiol and progesterone and cause anestrus in negative energy balance. These data provide a better understanding of the changes that may affect estrus of postpartum dairy cows at NEB status and lay the ground for further research.(AU)
O objetivo do presente estudo foi investigar os diferentes metabolitos do plasma entre o cio e o cio pós-parto de vacas leiteiras e fornecer uma base teórica para a prevenção do cio de vacas em fazendas de leite. No experimento, foram selecionadas 127 vacas leiteiras Holstein com idade e paridade similares. De acordo com a concentração de ß- ácido hidroxibutírico, ácidos graxos não esterificados e glicose no plasma entre 14 e 21 dias no leite, todas as vacas leiteiras foram determinadas em estado de equilíbrio energético. De acordo com os resultados dos sintomas clínicos, do exame de ultra-som retal e B aos 60 a 90 dias pós-parto, estas vacas foram divididas em vinte cios e vinte e quatro grupos de cio, outras vacas leiteiras foram removidas. A tecnologia de ressonância magnética nuclear 1H foi utilizada para detectar as alterações dos metabólitos plasmáticos e para triar diferentes metabólitos plasmáticos entre as vacas do cio e do cio. Dez diferentes metabólitos incluindo alanina, ácido glutâmico, asparagina, creatina, colina, fosfocholina, glicerofosfocolina, lipoproteína de baixa densidade e lipoproteína de muito baixa densidade foram significativamente diminuídos nas vacas antróficas em comparação com as vacas estro. As análises da via metabólica indicaram que os metabólitos diferenciais estavam principalmente envolvidos no metabolismo de aminoácidos e glicerofosfolipídios. Estes metabólitos e suas vias de enriquecimento indicam que a redução dos precursores da síntese de hormônios esteróides resulta em níveis mais baixos de estradiol e progesterona e causa anestros no balanço energético negativo. Estes dados fornecem uma melhor compreensão das mudanças que podem afetar o cio das vacas leiteiras pós-parto no estado de NEB e preparam o terreno para mais pesquisas.(AU)
Asunto(s)
Animales , Femenino , Bovinos , Progesterona/análisis , Anestro/sangre , Estro/sangre , Periodo Posparto/sangre , Estradiol/análisis , Glicerofosfolípidos , Ácidos Grasos no Esterificados , Aminoácidos , Glucosa , Pruebas Hematológicas/veterinariaRESUMEN
The objective of this study was to investigate the influence of plasma pathological changes before timed artificial insemination (TAI) on pregnancy of cows. The contents of estrogen (E2), progesterone (P4), glucose (Glu), selenium (Se), brain-derived neurotrophic factor (BDNF), and histamine (HIS) in plasma of 48 Holstein cows were measured before TAI. According to the estrus detection, the cows were divided into estrus (E) and anestrus (A) groups. After pregnancy testing at 28 d after TAI, two groups of E and A were divided into positive pregnancy of E group (EP+), negative pregnancy of E group (EP-), positive pregnancy of A group (AP+), and negative pregnancy of A group (AP-). The contents of E2, P4, Glu, Se, BDNF and hIS significantly differed among the four groups (P<0.01). The ROC analysis was used to determine the risk of negative pregnancy test (-) after TAI was increased when plasma E2 was less than 46.45 pmol/L in cows before TAI. The changes in E2, P4,hIS, Glu, and BDNF in the blood of natural estrus and natural anestrus cows affected the pregnancy after TAI. the level of E2 in plasma may be used to assess the risk of negative pregnancy after TAI.(AU)
O objetivo do presente estudo foi investigar a influência de mudanças patológicas de plasma antes de inseminação artificial (TAI) na gestação de vacas. O conteúdo de estrogênio (E2), progesterona (P4), glucose (Glu), selênio (Se), fator neurotrófico derivado do cérebro (BDNF), e histamina (HIS) no plasma de 48 vacas Holstein foi medido antes de TAI. De acordo com a detecção de estro, as vacas foram divididas em dois grupos: estro (E) e anestro (A). Após teste de gestação 28 d após TAI, dois grupos de E e A foram formados em gestação positiva do grupo E (EP+), gestação negativa do grupo E (EP-), gestação positiva do grupo A (AP+), e gestação negativa do grupo A (AP-). Os valores de E2, P4, Glu, Se, BDNF e hIS foram significativamente diferentes entre os quatro grupos (P<0,01). A análise ROC foi utilizada para determinar o risco de teste de gestação negativo (-) após aumento de TAI quando plasma E2 estava abaixo de 46,45 pmol/L em vacas antes de TAI. Alterações em E2, P4,hIS, Glu e BDNF no sangue de estro natural e anestro natural em vacas afetou a gestação após TAI. O nível de E2 no plasma pode ser usado para avaliar o risco de gestação negativa após TAI.(AU)
Asunto(s)
Animales , Femenino , Embarazo , Bovinos , Plasma , Anestro/sangre , Estro/sangre , Inseminación Artificial/veterinaria , Curva ROCRESUMEN
BACKGROUND: Although the 5-year survival rates in pediatric acute myeloid leukemia (AML) have improved over the last decades, there is a high relapse rate for Pediatric AML patients. METHODS: In the present study, we mainly combine PCA with the LASSO technique to identify prognostic markers for Pediatric AML patients coming from the NCI TARGET database. RESULTS: Three key genes (EEF1A1, RPLP2, RPL19) associated with poor prognosis of pediatric AML has been screened by both PCA and LASSO Cox regression analysis. Simultaneously, we developed a risk score model to predict the prognosis of pediatric AML, according to risk scores, the patients were divided into high- and low-risk groups based on the median risk score. Kaplan-Meier survival analysis indicated that Pediatric AML patients with the high-risk group have a poorer survival rate than those with a low-risk group (p < 0.000). The receiver operating characteristic (ROC) analysis showed that the risk model has a good performance (AUC:0.669). Moreover, the clinicopathologic correlation showed that the expression levels of three genes were related to the central nervous system (CNS) disease and chloroma. GSEA identified that those pathways including oxidative phosphorylation, apoptosis and TGFB signaling pathway were differentially enriched. CONCLUSION: Taken together, those studies suggested that a gene panel that consists of three genes (EEF1A1, RPLP2, RPL19) may act as a potential prognostic marker.
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Biomarcadores de Tumor/genética , Leucemia Mieloide Aguda/genética , Factor 1 de Elongación Peptídica/genética , Fosfoproteínas/genética , Proteínas Ribosómicas/genética , Apoptosis , Área Bajo la Curva , Neoplasias del Sistema Nervioso Central/genética , Niño , Bases de Datos Factuales , Proteínas de Homeodominio , Humanos , Estimación de Kaplan-Meier , Leucemia Mieloide Aguda/mortalidad , Fosforilación Oxidativa , Pronóstico , Modelos de Riesgos Proporcionales , Curva ROC , Proteínas Represoras , Factores de Riesgo , Sarcoma Mieloide/genética , Tasa de SupervivenciaRESUMEN
INTRODUCTION: Urothelial carcinoma (UC) is an aggressive malignancy and has a poor prognosis in the metastatic state. Treatment of UC remains a challenge, and as a first-line regimen for advanced UC, standard platinum-based chemotherapy is unfit for many patients due to numerous comorbidities and poor performance status. Recently, five immune checkpoint inhibitors have been approved for the treatment of patients with advanced UC who were ineligible for platinum-based regimens or suffered tumor progression in post-platinum setting. However, not long ago, the U.S. Food and Drug Administration restricted the use of two common immune checkpoint blockades, atezolizumab and pembrolizumab, due to uncertain survival benefit as mono-therapy. In this scenario, we reviewed rapidly surfacing clinical trials to assess the efficacy and safety of immunotherapy targeting the PD-1 pathway for advanced UC. METHODS: A comprehensive search was conducted in PubMed, EMBASE and Cochrane Library for all clinical trials where the efficacy and safety were reported. Our primary outcome was efficacy evaluated by objective response rate (ORR), 1-year overall survival (OS) rate and 1-year progression-free survival (PFS) rate, and second outcome was safety assessed by any grade and grade 3-4 treatment-related adverse events (TRAEs). We chose percentages with 95% confidence intervals (CI) as the evaluation indexes and used a random-effects model to account for heterogeneity. RESULTS: We included 14 clinical trials with 2674 total patients in this meta-analysis. After removing unqualified studies on the basis of sensitivity analyses, 13 studies were pooled to evaluate the overall ORR, 8 studies for the 1-year OS rate and 6 studies for the 1-year PFS rate. The pooled data of ORR, 1-year OS rate, and 1-year PFS rate were 0.20 (95% CI 0.18-0.22, I2 = 38.4%, P = 0.078), 0.50 (95% CI 0.46-0.53, I2 = 30.3%, P = 0.186), and 0.17 (95% CI 0.14-0.20, I2 = 0.0%, P = 0.668), respectively. Similarly, 13 trials were utilized to compute the pooled rate of any-grade TRAEs. The pooled estimation of any-grade was 0.65 (95% CI 0.63-0.67, I2 = 1.7%, P = 0.429). The pooled rate of grade 3-4 TRAEs subgroups with Atezolizumab, Pembrolizumab, Durvalumab, Nivolumab and Avelumab were 0.11 (95% CI 0.06-0.15, I2 = 83.5%, P = 0.000), 0.15 (95% CI 0.13-0.18, I2 = 0.0%, P = 0.971), 0.06 (95% CI 0.03-0.09, I2 = 0.0%, P = 0.566), 0.19 (95% CI 0.15-0.23, I2 = 0.0%, P = 0.480) and 0.08 (95% CI 0.05-0.11, I2 = 0.0%, P = 0.702), respectively. CONCLUSION: This study showed that the immunotherapy targeting the PD-1 pathway had durable efficacy and acceptable safety in patients with advanced UC. The comprehensive role of immune checkpoint inhibitors in comparison to other treatments needs further confirmation basing on RCTs.
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Inhibidores de Puntos de Control Inmunológico/uso terapéutico , Neoplasias Urológicas/tratamiento farmacológico , Urotelio/patología , Ensayos Clínicos como Asunto , Humanos , Inhibidores de Puntos de Control Inmunológico/efectos adversos , Sesgo de Publicación , Neoplasias Urológicas/mortalidadRESUMEN
BACKGROUND: T cell therapy for cancer involves genetic introduction of a target-binding feature into autologous T cells, ex vivo expansion and single large bolus administration back to the patient. These reprogrammed T cells can be highly effective in killing cells, but tumor heterogeneity results in regrowth of cells that do not sufficiently express the single antigen being targeted. We describe a cell-based therapy that simultaneously targets multiple tumor-specific antigens. METHODS: High-affinity polyclonal rabbit antibodies were generated against nine different surface-related tumor-specific mutations on B16F10 cells. Unsorted splenic effector cells from syngeneic mice were incubated with a cocktail of the nine anti-B16F10 antibodies. These 'armed' effector cells were used to treat mice previously inoculated with B16F10 melanoma cells. RESULTS: The cocktail of nine antibodies resulted in dense homogeneous binding to histological sections of B16F10 cells. Five treatments with the armed effector cells and PD1 inhibition inhibited tumor growth and improved survival. Shortening the interval of the five treatments from every three days to every day increased survival. Arming effector cells with the four antibodies showing best binding to B16F10 cells even further increased survival. CONCLUSIONS: This study demonstrates that ex vivo arming a mixed population of immune effector cells with antibodies targeting multiple tumor-specific mutated proteins in conjunction with PD1 inhibition delayed tumor growth and prolonged survival in mice inoculated with an aggressive melanoma. A remarkably low total antibody dose of less than 5 µg was sufficient to accomplish tumor inhibition. Scaling up to clinical level may be feasible.
Asunto(s)
Anticuerpos Antineoplásicos/uso terapéutico , Antígenos de Neoplasias/inmunología , Inmunoterapia Adoptiva/métodos , Leucocitos/inmunología , Melanoma Experimental/terapia , Neoplasias Cutáneas/terapia , Carga Tumoral , Animales , Antígenos de Neoplasias/genética , Melanoma Experimental/inmunología , Melanoma Experimental/patología , Ratones , Proteínas Mutantes/genética , Proteínas Mutantes/inmunología , Mutación , Receptor de Muerte Celular Programada 1/antagonistas & inhibidores , Neoplasias Cutáneas/inmunología , Neoplasias Cutáneas/patología , Bazo/citología , Tasa de SupervivenciaRESUMEN
BACKGROUND: Angiogenesis is an indispensable step in the growth and invasiveness of breast cancers involving a series of exquisite molecular steps. Pro-angiogenic factors, including vascular endothelial growth factor (VEGF), have been recognized as pivotal therapeutic targets in the treatment of breast cancer. More recently, a highly conserved transcription factor Twist has been reported to be involved in tumor angiogenesis and metastasis. METHODS: The expression of VEGF-C and Twist was immunohistochemically determined in tissue samples of primary tumors from 408 patients undergoing curative surgical resection for breast cancer. The correlations of VEGF-C and Twist expressions with clinicopathologic parameters as well as survival outcomes were evaluated. RESULTS: Of the 408 patients evaluated, approximately 70% had high expression of VEGF-C which was significantly associated with advanced tumor stages (P = 0.019). Similarly, VEGF-C expression was associated with the proliferation index Ki67, N3 lymph node metastasis, and D2-40-positive lymphatic vessel invasion (LVI) in a univariate analysis. Furthermore, patients with high expressions of VEGF-C and Twist (V + T+) had significantly increased lymph node metastasis, higher clinical stage, and worse disease-free survival, DFS (P = 0.001) and overall survival, OS (P = 0.011). CONCLUSIONS: Our results suggested that co-expression of VEGF-C and Twist was associated with larger tumor size, higher numbers of lymph node involvement, D2-40-positive LVI, higher risk of distant metastasis, and worse DFS or OS in breast cancer patients.
Asunto(s)
Biomarcadores de Tumor/metabolismo , Neoplasias de la Mama/patología , Carcinoma Ductal de Mama/patología , Proteínas Nucleares/metabolismo , Proteína 1 Relacionada con Twist/metabolismo , Factor C de Crecimiento Endotelial Vascular/metabolismo , Neoplasias de la Mama/metabolismo , Neoplasias de la Mama/cirugía , Carcinoma Ductal de Mama/metabolismo , Carcinoma Ductal de Mama/cirugía , Femenino , Estudios de Seguimiento , Regulación Neoplásica de la Expresión Génica , Humanos , Persona de Mediana Edad , Pronóstico , Tasa de SupervivenciaRESUMEN
BACKGROUND: The subgroup of young patients with non-small-cell lung cancer (NSCLC) is poorly understood. We retrospectively studied the clinical characteristics, gene mutations, and outcomes of patients with NSCLC (aged ≤ 40 years). RESULTS: Of the 7494 patients with lung cancer diagnosed from February 2001 to October 2016, 252 aged ≤ 40 years showed NSCLC. We divided their cases into non-squamous cell carcinoma and squamous cell carcinoma groups according to their histology results. Of the 252 young NSCLC patients, 173 (69%) patients had stage IIIB or IV, and 196 (78%) had never smoked. The four most common metastases were intrapulmonary lesions, pleura, bone, and brain. Among patients with adenocarcinoma, 29 (40%, n = 73) harbored epidermal growth factor receptor (EGFR) mutations, 25 (34%, n = 74) harbored anaplastic lymphoma kinase (ALK) translations, and 1 (14%, n = 7) harbored ROS proto-oncogene 1 receptor tyrosine kinase (ROS1) translations. The median progression-free survival (PFS) and overall survival (OS) were 3.3 and 27.6 months for patients receiving chemotherapy (n = 65), and 12.1 and 33.6 months for patients receiving EGFR tyrosine kinase inhibitors (TKIs) (n = 13), respectively. Patients receiving crizotinib had a median PFS time of 21.9 months (n = 8). CONCLUSIONS: Young patients are associated with an increased likelihood of gene mutations and can receive a better prognosis when patients harboring gene mutations are treated with EGFR-TKIs or ALK inhibitors.
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Carcinoma de Pulmón de Células no Pequeñas/genética , Neoplasias Pulmonares/genética , Mutación , Adulto , Quinasa de Linfoma Anaplásico , Carcinoma de Pulmón de Células no Pequeñas/tratamiento farmacológico , Carcinoma de Pulmón de Células no Pequeñas/mortalidad , Carcinoma de Pulmón de Células no Pequeñas/secundario , Receptores ErbB/genética , Femenino , Humanos , Neoplasias Pulmonares/tratamiento farmacológico , Neoplasias Pulmonares/mortalidad , Neoplasias Pulmonares/patología , Masculino , Pronóstico , Proto-Oncogenes Mas , Proteínas Tirosina Quinasas Receptoras/genética , Estudios RetrospectivosRESUMEN
BACKGROUND: Radioiodine therapy (RAI) after total or near-total thyroidectomy is a recommended treatment for patients with pulmonary metastasis from differentiated thyroid cancer (DTC). However, the total effective rate of iodine-131 therapy remains controversial. This study aimed to determine the efficacy of RAI for treating patients with pulmonary metastasis from DTC, and to identify independent predictors of its efficacy. METHODS: We conducted a retrospective study to evaluate 20 patients with pulmonary metastasis from DTC who underwent RAI at our center at first and performed a meta-analysis to evaluate relevant literature regarding the overall efficacy of RAI and subgroup-specific efficacies subsequently. RESULTS: The efficacy rate at our center was 40%, and no significant differences were observed according to sex, age, pathological type, metastasis state, or interval between the initial RAI and final surgery. The meta-analysis revealed that the pooled overall efficacy rate was 58%, and significant differences were observed when we compared pulmonary metastasis versus pulmonary and other distant metastasis, age of < 40 years versus age of ≥ 40 years, papillary thyroid cancer versus follicular thyroid cancer and male patients versus female patients. CONCLUSIONS: These results suggest that RAI is an effective treatment for patients with pulmonary metastasis from DTC after surgical treatment. The efficacy of RAI was significantly predicted by the presence of papillary thyroid cancer, age of < 40 years, the absence of non-lung distant metastasis and female patients.
Asunto(s)
Adenocarcinoma Folicular/radioterapia , Neoplasias Óseas/radioterapia , Carcinoma Papilar/radioterapia , Radioisótopos de Yodo/uso terapéutico , Neoplasias Pulmonares/radioterapia , Neoplasias de la Tiroides/radioterapia , Adenocarcinoma Folicular/patología , Adulto , Anciano , Neoplasias Óseas/secundario , Carcinoma Papilar/patología , Femenino , Estudios de Seguimiento , Humanos , Neoplasias Pulmonares/secundario , Metástasis Linfática , Masculino , Metaanálisis como Asunto , Persona de Mediana Edad , Pronóstico , Estudios Retrospectivos , Tasa de Supervivencia , Neoplasias de la Tiroides/patología , Adulto JovenRESUMEN
Adipose-derived stem cells (ADSCs) show nearly unlimited potential in medical and animal science. Currently, understanding of the biological mechanisms regulating ADSC growth in vitro remains very limited. Histone acetylation, an epigenetic modification, plays a key role in maintaining stem cell properties. To further study its effect on ADSC growth characteristics in vitro, we treated goat ADSCs with the histone deacetylase inhibitors trichostatin A (TSA) and vorinostat (SAHA). This inhibited SIRT1 expression and increased histone H3K9 acetylation, leading to decreased cell viability, cell cycle arrest, and apoptosis. Quantitative real-time polymerase chain reaction revealed that H3K9 hyperacetylation stimulated transcription of NANOG, OCT4, SOX2, and TERT, but inhibited that of PCNA, P53, and BAX. Western blotting indicated that TSA and SAHA increased protein expression of NANOG, reduced that of SOX2, TERT, PCNA, P53, and BAX, and did not change that of OCT4. These findings provide new experimental evidence contributing to our understanding of the mechanisms underlying ADSC growth characteristics in vitro.
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Tejido Adiposo/citología , Histonas/metabolismo , Células Madre/citología , Células Madre/metabolismo , Acetilación , Animales , Apoptosis/genética , Western Blotting , Ciclo Celular/genética , Proliferación Celular , Supervivencia Celular/genética , Citometría de Flujo , Regulación de la Expresión Génica , Cabras , Histona Desacetilasas/genética , Histona Desacetilasas/metabolismo , Reacción en Cadena de la Polimerasa , ARN Mensajero/genética , ARN Mensajero/metabolismoRESUMEN
The matrix Gla (gamma-carboxyglutamic acid-rich) protein (MGP), a vitamin K-dependent and Gla-containing protein, is a calcification inhibitor that mainly functions in tissue calcification and mineralization. In this study, we obtained the complete cDNA sequence of MGP from the spinyhead croaker (Collichthys lucidus), which we named Cl-MGP. Cl-MGP was 923 bp long with a 384-bp open reading fragment that encoded 127 amino acids. The predicted MGP protein sequence contained a 19-residue hydrophobic signal peptide, suggesting that it possesses secretory characteristics. The Gla domain and the invariant unit ErraEtCedyspC, which has been identified in all known vitamin K-dependent vertebrate proteins, were highly conserved in Cl-MGP, suggesting that it uses the same mechanism to function as the known proteins. An alignment analysis revealed that Cl-MGP had the highest identity with Larimichthys crocea (93%), which had lost five amino acid residues in the C-terminal. A quantitative real-time polymerase chain reaction revealed that Cl-MGP expression was highest in the gill, followed by the cholecyst and spleen, with almost no expression in the blood, muscle, or testes. The high Cl-MGP expression in the gill is similar to that observed in other fish species, but the relatively high expression found in the cholecyst and spleen is not seen in all species. Future studies should investigate the tissue distributions of both mRNA and proteins in different species, in order to understand the function and evolution of MGP in different species.
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Proteínas de Unión al Calcio/genética , Proteínas de la Matriz Extracelular/genética , Regulación de la Expresión Génica , Perciformes/genética , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Proteínas de Unión al Calcio/química , Proteínas de Unión al Calcio/metabolismo , Clonación Molecular , ADN Complementario/genética , Proteínas de la Matriz Extracelular/química , Proteínas de la Matriz Extracelular/metabolismo , Perfilación de la Expresión Génica , Especificidad de Órganos/genética , Filogenia , Alineación de Secuencia , Proteína Gla de la MatrizRESUMEN
Loss of function of mutated solute carrier family 12 member 3 (SLC12A3) gene is the most frequent etiology for Gitelman syndrome (GS), which is mainly manifested by hypokalemia, hypomagnesemia and hypocalciuria. We report the genetic characteristics of one suspicious Chinese GS pedigree by gene sequencing. Complete sequencing analysis of the SLC12A3 gene revealed that both the proband and his elder sister had a novel homozygous SLC12A3 mutation: c.2099T>C and p.Leu700Pro. Moreover, the SLC12A3 genes of his mother and daughter encoded the same mutated heterozygote. It was noted that in this pedigree, only the proband complained about recurrent episodes of bilateral lower limb weakness over 8 years, while his elder sister, mother and daughter did not present symptoms. The inconsistent clinical features of this pedigree implied that besides diverse phenotypes possibly originated from the same genotype, gender difference may also dominate the variant GS phenotypes. Further genetic and proteomic research are needed to investigate the precise mechanisms of GS, including the study of specific ethnicities.
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Síndrome de Gitelman/genética , Homocigoto , Mutación/genética , Miembro 3 de la Familia de Transportadores de Soluto 12/genética , Pueblo Asiatico , Femenino , Síndrome de Gitelman/diagnóstico , Humanos , Masculino , Linaje , Fenotipo , Adulto JovenRESUMEN
Hepatocellular carcinoma is one of the most common malignant tumors worldwide; it is estimated that there were 782,000 new cases in 2012. MicroRNAs (miRNAs) play an important role in carcinogenesis by regulating oncogenes and tumor suppressors. We investigated the role of miR-146a, miR-196a2, and miR-499 polymorphisms in the risk of hepatocellular carcinoma in a Chinese population. Hepatocellular carcinoma patients (175) and healthy controls (302) were recruited between April 2013 and March 2015. Genotype analysis of miR-146a, miR-196a2, and miR-499 polymorphisms was carried out by polymerase chain reaction-restriction fragment length polymorphism. There was a significant difference between the genotype distribution of miR-196a2 in hepatocellular carcinoma patients and controls (X2 = 17.23, P < 0.001). CG and GG miR-146a genotypes significantly elevated the risk of hepatocellular carcinoma compared with the CC genotype, with adjusted ORs (95%CI) of 3.05 (1.07-8.70) and 4.96 (1.64-14.97), respectively. In the recessive model, the CG + GG genotype had a 3.75-fold risk of hepatocellular carcinoma compared with the CC genotype, with an adjusted OR (95%CI) of 3.75 (1.39-10.11). However, no significant association was observed between miR-196a2 and miR-499 variants and risk of hepatocellular carcinoma in the co-dominant, dominant, and recessive models. The miR-146a polymorphism is a G to C substitution that causes a mismatch in the stem-loop of miRNA, which influences how the expression and transcriptional regulation of miRNA affects its target genes. Our study revealed that the GG and CG genotypes of miR-146a increased the risk of hepatocellular carcinoma in the Chinese population.
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Carcinoma Hepatocelular/genética , Neoplasias Hepáticas/genética , MicroARNs/genética , Polimorfismo de Nucleótido Simple , Anciano , Estudios de Casos y Controles , Femenino , Frecuencia de los Genes , Genotipo , Humanos , Masculino , Persona de Mediana EdadRESUMEN
Diabetic retinopathy (DR), an important complication of diabetes mellitus (DM), is not well understood. T helper cell balance (Th1/Th2) is involved in various autoimmune diseases; however, its role in DR is not understood. This study explores changes in Th1 and Th2 cytokine expression during DR. Blood samples were collected from 25 healthy volunteers (normal control group), 35 patients with type 2 DM (T2DM group) without DR, and 30 cases of T2DM patients with DR (DR group). Real-time PCR was used to measure mRNA expression of IL-2 and TNF-α, secreted from Th1 cells, and of IL-4 and IL-10, secreted from Th2 cells. We used ELISA to detect cytokine expression in serum to analyze the correlation between Th1 and Th2 cytokines. IL-2 and TNF-αmRNA and protein expression levels in the T2DM and DR groups were significantly higher than in the normal control group (P < 0.05). Compared with the T2DM group, the DR group had higher IL-2 and TNF-αlevels (P < 0.05). IL-4 and IL-10 levels were lower in the DR group compared with the normal and T2DM groups (P < 0.05), while T2DM showed no difference compared with the normal control (P > 0.05). IL-2 and TNF-αwere negatively correlated with IL-4 and IL-10 in the DR group, respectively. We found that Th1 cytokine secretion was higher and Th2 cytokines secretion was lower during DR, leading to a Th1/ Th2 imbalance, suggesting that Th1/Th2 imbalance is a side effect for DR occurrence and development.
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Citocinas/sangre , Diabetes Mellitus Tipo 2/sangre , Retinopatía Diabética/sangre , Células TH1/metabolismo , Células Th2/metabolismo , Adulto , Anciano , Biomarcadores/sangre , Estudios de Casos y Controles , Diabetes Mellitus Tipo 2/complicaciones , Retinopatía Diabética/etiología , Femenino , Humanos , Masculino , Persona de Mediana EdadRESUMEN
Populus cathayana occupies a large area within the northern, central, and southwestern regions of China, and is considered to be an important reforestation species in western China. In order to investigate the population genetic structure of this species, 10 polymorphic microsatellite loci were identified based on expressed sequence tags from de novo sequencing on the Illumina HiSeq 2000 platform. All microsatellite primers were tested on 48 P. cathayana individuals from four locations on the Qinghai-Tibet Plateau. The observed heterozygosity ranged from 0.000 to 1.000, and the null-allele frequency ranged from 0.000 to 0.904. These microsatellite markers may be a useful tool in genetic studies on P. cathayana and closely related species.
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Etiquetas de Secuencia Expresada , Repeticiones de Microsatélite , Polimorfismo Genético , Populus/genética , Frecuencia de los Genes , Marcadores Genéticos , HeterocigotoRESUMEN
We investigated the effect of microRNA-27b (miR-27b), a gene expression regulatory factor, on the expression of monocyte chemoattractant protein-1 (MCP1) stimulated by interleukin 17 (IL-17). After IL-17 had been added to H9C2 cardiomyocytes, an miR-27b mimic was transfected into the H9C2 cells. The mRNA expression levels of miR-27b and MCP1 in the H9C2 cells were detected by SYBR green I fluorescence quantitative reverse transcription polymerase chain reaction. Enzyme-linked immunosorbent assay was used to measure the expression levels of MCP1 protein in the H9C2 cells. The expression of MCP1 mRNA in the H9C2 cells began to increase 2 h after IL-17 stimulation, reached a peak at 4 h, and then decreased. The MCP1 protein level increased gradually in the 24 h following IL-17 stimulation. After transfection with the miR-27b mimic, the expression of miR-27b in the H9C2 cells significantly increased than that in the miRNA negative control group (P < 0.01). The MCP1 mRNA and protein levels in the miR-27b mimic + IL-17 group were significantly reduced than that in the miRNA negative control + IL-17 group (P < 0.01). miR-27b inhibited IL-17-induced MCP1 expression in the H9C2 cells, which demonstrates that treatment with microRNA could alleviate myocardial injury in viral myocarditis.
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Quimiocina CCL2/biosíntesis , Interleucina-17/antagonistas & inhibidores , MicroARNs/administración & dosificación , Animales , Células Cultivadas , Quimiocina CCL2/genética , Regulación de la Expresión Génica , Interleucina-17/farmacología , MicroARNs/biosíntesis , MicroARNs/genética , Miocardio/citología , Miocardio/metabolismo , Miocitos Cardíacos/efectos de los fármacos , Miocitos Cardíacos/metabolismo , ARN Mensajero/biosíntesis , ARN Mensajero/genética , ARN Mensajero/metabolismo , Ratas , Transducción de Señal , Transfección/métodosRESUMEN
Cystatins are natural tight-binding reversible inhibitors of cysteine proteases. In this study, a cDNA library was constructed from Collichthys lucidus using the SMART technique. A complete cDNA sequence with high identity to the conserved sequence of the cystatin C gene was cloned from the library using EST analysis and rapid amplification of cDNA ends (RACE), then subjected to further investigation. The full-length cDNA of cystatin C from C. lucidus (Clcys) was 699 bp long, including a 5'-terminal untranslated region (5'-UTR) of 52 bp, a 3'-UTR of 290 bp, and an open-reading frame of 357 bp. The gene encoded a polypeptide of 118 amino acids, constituting a predicted molecular weight of 12.875 kDa and a theoretical isoelectric point of 8.81. The amino acid sequence of Clcys possessed typical features of type II cystatins and had the highest identity with cystatin C of Pseudosciaena crocea (89%); therefore, it clustered with the cystatin C group in the UPGMA phylogenetic tree. Quantitative real-time reverse transcription analysis revealed that the highest expression was found in the kidney, followed by the liver, heart, and testis, with the lowest expression in muscle. Interestingly, Clcys had relatively low identity with cystatin C genes from other fish and mammals, and its expression pattern did not possess features of a housekeeping gene. Based on these findings, we suspect that the classification of cystatins in fish is somewhat confusing, and the identification of more cystatin gene sequences is needed before a definite conclusion can be drawn.
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Cistatina C/genética , Cistatina C/metabolismo , ADN Complementario/genética , Animales , Clonación Molecular , Perfilación de la Expresión Génica , Sistemas de Lectura Abierta/genética , PerciformesRESUMEN
PURPOSE: The aim of this study was (1) to evaluate and predict the value of ProGRP and NSE in therapy and survival; (2) as well as to investigate the correlation between the ProGRP mRNA expression in peripheral blood and serum ProGRP protein. METHODS: The study included 122 patients with SCLC without prior therapy. The serum levels of ProGRP and NSE were detected by enzyme-linked immunosorbent assay and eletro-chemiluminescence immunoassay, respectively. The expression of ProGRP mRNA was detected by real-time reverse transcriptase-polymerase chain reaction. RESULTS: Distribution of serum levels of ProGRP, NSE and ProGRP mRNA differed significantly according to tumor size, disease stage and distant metastasis (all P < 0.05), and no association was found between them and gender or age (both P > 0.05). After two courses of chemotherapy, patients of remission and stable groups showed a marked decrease in ProGRP and NSE concentrations (P < 0.05). The ProGRP concentration of patients in progression group was significantly higher than pretreatment level (P < 0.05), while NSE concentration was not. A linear nonparametric (Spearman) correlation test revealed that there was a significant correlation between ProGRP mRNA expression in peripheral blood and serum ProGRP protein level (P < 0.05). Univariate analysis found a statistically significant association of survival with disease stage, distant metastasis, ProGRP and NSE (P < 0.05). Gender, age and tumor size were not prognostic factors (P > 0.05). Multiple Cox regression model analysis found that only disease stage and NSE were significant predictors (P < 0.05). CONCLUSIONS: This study has found that there is a potential role for ProGRP and NSE in both therapy monitoring and predicting survival in SCLC patients.
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Biomarcadores de Tumor/sangre , Quimioradioterapia/mortalidad , Neoplasias Pulmonares/mortalidad , Fragmentos de Péptidos/sangre , Fosfopiruvato Hidratasa/sangre , Carcinoma Pulmonar de Células Pequeñas/mortalidad , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Biomarcadores de Tumor/genética , Técnicas Electroquímicas , Ensayo de Inmunoadsorción Enzimática , Femenino , Estudios de Seguimiento , Humanos , Inmunoensayo , Mediciones Luminiscentes , Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/patología , Metástasis Linfática , Masculino , Persona de Mediana Edad , Estadificación de Neoplasias , Fragmentos de Péptidos/genética , Fosfopiruvato Hidratasa/genética , Reacción en Cadena de la Polimerasa , Pronóstico , Proteínas Recombinantes/sangre , Proteínas Recombinantes/genética , Carcinoma Pulmonar de Células Pequeñas/metabolismo , Carcinoma Pulmonar de Células Pequeñas/secundario , Tasa de SupervivenciaRESUMEN
We aimed to evaluate the effectiveness and safety of bismuth-containing quadruple therapy plus postural change after dosing for Helicobacter pylori eradication in gastrectomized patients. We compared 76 gastric stump patients with H. pylori infection (GS group) with 50 non-gastrectomized H. pylori-positive patients who met the treatment indication (controls). The GS group was divided into GS group 1 and GS group 2. All groups were administered bismuth potassium citrate (220 mg), esomeprazole (20 mg), amoxicillin (1.0 g), and furazolidone (100 mg) twice daily for 14 days. GS group 1 maintained a left lateral horizontal position for 30 min after dosing. H. pylori was detected using rapid urease testing and histologic examination of gastric mucosa before and 3 months after therapy. Mucosal histologic manifestations were evaluated using visual analog scales of the updated Sydney System. GS group 1 had a higher prevalence of eradication than the GS group 2 (intention-to-treat [ITT]: P=0.025; per-protocol [PP]: P=0.030), and the control group had a similar prevalence. GS group 2 had a lower prevalence of eradication than controls (ITT: P=0.006; PP: P=0.626). Scores for chronic inflammation and activity declined significantly (P<0.001) 3 months after treatment, whereas those for atrophy and intestinal metaplasia showed no significant change. Prevalence of adverse reactions was similar among groups during therapy (P=0.939). A bismuth-containing quadruple therapy regimen plus postural change after dosing appears to be a relatively safe, effective, economical, and practical method for H. pylori eradication in gastrectomized patients.