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1.
Biol Reprod ; 2024 Jun 20.
Artigo em Inglês | MEDLINE | ID: mdl-38900909

RESUMO

Cytoplasmic dynein participates in transport functions and is essential in spermatogenesis. KM23 belongs to the dynein light chain family. The TGFß signaling pathway is indispensable in spermatogenesis, and Smad2 is an important member of this pathway. We cloned PTKM23 and PTSMAD2 from Portunus trituberculatus and measured their expression during spermatogenesis. PTKM23 may be related to cell division, acrosome formation and nuclear remodeling, and PTSMAD2 may participate in regulating the expression of genes related to spermatogenesis. We assessed the localization of PTKM23 with PTDHC and α-Tubulin, and the results suggested that PTKM23 functions in intracellular transport during spermatogenesis. We knocked down PTKM23 in vivo, and the expression of p53, B-CATAENIN and CYCLIN B decreased significantly, further suggesting a role of PTKM23 in transport and cell division. The localization of PTDIC with α-Tubulin and that of PTSMAD2 with PTDHC changed after PTKM23 knockdown. We transfected PTKM23 and PTSMAD2 into HEK-293 T cells and verified their colocalization. These results indicate that PTKM23 is involved in the assembly of cytoplasmic dynein and microtubules during spermatogenesis and that PTKM23 mediates the participation of cytoplasmic dynein in the transport of PTSMAD2 during spermatogenesis. This study provides a theoretical molecular biological basis for the breeding of P. trituberculatus.

2.
Fish Shellfish Immunol ; 145: 109355, 2024 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-38168634

RESUMO

The scavenger receptor class B family proteins (SRB) are multiligand membrane receptor proteins. Herein, a novel SRB homolog (Pt-SRB2) was identified in Portunus trituberculatus. The open reading frame of Pt-SRB2 was predicted to encode 520 amino acid residues comprising a typical CD36 domain. Phylogenetic analysis showed that Pt-SRB2 distinctly clustered with the SRB homologs of most crustaceans and Drosophila but was separate from all vertebrate CD36/SRB. Semi-quantitative and Real-time quantitative PCR revealed that the abundance of Pt-SRB2 transcripts was the highest in hepatopancreas than in other tested tissues. Overexpressed Pt-SRB2 was distributed primarily in the cell membrane and cytoplasm of HEK293T or Drosophila Schneider 2 cells. In crab hemocytes, Pt-SRB2 was distributed primarily in the cell membrane by immunofluorescence staining. In addition, the immunofluorescence staining showed that green fluorescence signals were mainly located in the inner lumen membrane of the hepatopancreatic tubules. Moreover, solid-phase enzyme-linked immunosorbent assay revealed that rPt-SRB2-L exhibited relative high affinity with lipopolysaccharides, and relative moderate binding affinity with lipoteichoic acid or peptidoglycan. Of note, rPt-SRB2-L showed high binding affinity with eicosapentaenoic acid among a series of long-chain polyunsaturated fatty acids. Taken together, this study provided valuable data for understanding the functions of the crab CD36/SRB.


Assuntos
Braquiúros , Antígenos CD36 , Humanos , Animais , Antígenos CD36/genética , Braquiúros/genética , Sequência de Aminoácidos , Sequência de Bases , Filogenia , Células HEK293 , Drosophila/metabolismo
3.
Lipids Health Dis ; 23(1): 83, 2024 Mar 20.
Artigo em Inglês | MEDLINE | ID: mdl-38509578

RESUMO

OBJECTIVE: To enhance the detection, management and monitoring of Chinese children afflicted with sitosterolemia by examining the physical characteristics and genetic makeup of pediatric patients. METHODS: In this group, 26 children were diagnosed with sitosterolemia, 24 of whom underwent genetic analysis. Patient family medical history, physical symptoms, tests for liver function, lipid levels, standard blood tests, phytosterol levels, cardiac/carotid artery ultrasounds, fundus examinations, and treatment were collected. RESULTS: The majority (19, 73.1%) of the 26 patients exhibited xanthomas as the most prevalent manifestation. The second most common symptoms were joint pain (7, 26.9%) and stunted growth (4, 15.4%). Among the 24 (92.3%) patients whose genetics were analyzed, 16 (66.7%) harbored ABCG5 variants (type 2 sitosterolemia), and nearly one-third (8, 33.3%) harbored ABCG8 variants (type 1 sitosterolemia). Additionally, the most common pathogenic ABCG5 variant was c.1166G > A (p.Arg389His), which was found in 10 patients (66.7%). Further analysis did not indicate any significant differences in pathological traits among those carrying ABCG5 and ABCG8 variations (P > 0.05). Interestingly, there was a greater abundance of nonsense variations in ABCG5 than in ABCG8 (P = 0.09), and a greater frequency of splicing variations in ABCG8 than ABCG5 (P = 0.01). Following a change in diet or a combination of ezetimibe, the levels of cholesterol and low-density lipoprotein were markedly decreased compared to the levels reported before treatment. CONCLUSION: Sitosterolemia should be considered for individuals presenting with xanthomas and increased cholesterol levels. Phytosterol testing and genetic analysis are important for early detection. Managing one's diet and taking ezetimibe can well control blood lipids.


Assuntos
Hipercolesterolemia , Enteropatias , Erros Inatos do Metabolismo Lipídico , Fitosteróis , Fitosteróis/efeitos adversos , Xantomatose , Humanos , Criança , Lipoproteínas/genética , Membro 5 da Subfamília G de Transportadores de Cassetes de Ligação de ATP/genética , Fitosteróis/genética , Colesterol , Ezetimiba/uso terapêutico
4.
Zhongguo Dang Dai Er Ke Za Zhi ; 26(2): 118-123, 2024 Feb 15.
Artigo em Zh | MEDLINE | ID: mdl-38436307

RESUMO

Short stature in puberty significantly affects the physical and mental health of adolescents. The continuous acceleration of skeletal maturation, caused by sex hormones during puberty, limits the time available for growth and poses a considerable challenge for the treatment of short stature. To date, there is still no standardized treatment protocol for this disorder. However, puberty is the last period to improve the final adult height. Currently, commonly used pharmacological treatments in clinical settings include recombinant human growth hormone, gonadotropin-releasing hormone analogs, and third-generation aromatase inhibitors. In recent years, personalized treatment aiming to improve the final adult height has become a key focus in clinical practice. This article provides a comprehensive summary of research on pharmacological therapies for height improvement in pubertal children with short stature, offering valuable insights for healthcare professionals.


Assuntos
Nanismo , Hormônio do Crescimento Humano , Adolescente , Adulto , Criança , Humanos , Hormônio do Crescimento Humano/uso terapêutico , Pessoal de Saúde
5.
Fish Shellfish Immunol ; 140: 108940, 2023 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-37442309

RESUMO

The family of TRIM proteins with E3 ubiquitin ligase activity served important roles in the regulation of innate immune processes, in particular antiviral and proinflammatory cytokine responses. In this study, a novel TRIM37 homolog was identified from Penaeus monodon (named PmTRIM37). The PmTRIM37 protein contained three conserved domains (one RING finger domain, a B-box, and one Coiled-coil region) at its N-terminal and one Meprin and MATH domain at its C-terminal. The MATH domain was the characteristic of TRIM37 family. PmTRIM37 has relatively high expression in immune-related tissues such as hepatopancreas, gills, lymphoid organs and hemocytes. The expression levels of PmTRIM37 in hepatopancreas and lymphoid organs were significantly up-regulated after white spot syndrome virus (WSSV) infection. Knock down of PmTRIM37 promoted WSSV replication and VP28 expression, suggesting that PmTRIM37 played a negative role in WSSV infection. Further studies revealed that PmTRIM37 positively regulated the NF-κB pathway and Antimicrobial peptides (AMP) expression during WSSV infection. These findings indicated that PmTRIM37 might restrict WSSV replication by positively regulating NF-κB pathway during WSSV infection in P. monodon.


Assuntos
Penaeidae , Vírus da Síndrome da Mancha Branca 1 , Animais , NF-kappa B/metabolismo , Ubiquitina-Proteína Ligases/genética , Ubiquitina-Proteína Ligases/metabolismo , Proteínas com Motivo Tripartido/genética , Proteínas com Motivo Tripartido/metabolismo , Vírus da Síndrome da Mancha Branca 1/fisiologia
6.
Biomed Eng Online ; 22(1): 23, 2023 Mar 10.
Artigo em Inglês | MEDLINE | ID: mdl-36894984

RESUMO

BACKGROUND: Exercise electrocardiography (ECG) is a noninvasive test aiming at producing ischemic changes. However, resting ECG cannot be adopted in diagnosing myocardial ischemia till ST-segment depressions. Therefore, this study aimed to detect myocardial energy defects in resting ECG using the Hilbert-Huang transformation (HHT) in patients with angina pectoris. METHODS: Electrocardiographic recordings of positive exercise ECG by performing coronary imaging test (n = 26) and negative exercise ECG (n = 47) were collected. Based on the coronary stenoses severity, patients were divided into three categories: normal, < 50%, and ≥ 50%. During the resting phase of the exercise ECG, all 10-s ECG signals are decomposed by HHT. The RT intensity index, composed of the power spectral density of the P, QRS, and T components, is used to estimate the myocardial energy defect. RESULTS: After analyzing the resting ECG using HHT, the RT intensity index was significantly higher in patients with positive exercise ECG (27.96%) than in those with negative exercise ECG (22.30%) (p < 0.001). In patients with positive exercise ECG, the RT intensity index was gradually increasing with the severity of coronary stenoses: 25.25% (normal, n = 4), 27.14% (stenoses < 50%, n = 14), and 30.75% (stenoses ≥ 50%, n = 8). The RT intensity index of different coronary stenoses was significantly higher in patients with negative exercise ECG, except for the normal coronary imaging test. CONCLUSIONS: Patients with coronary stenoses had a higher RT index at the resting stage of exercise ECG. Resting ECG analyzed using HHT could be a method for the early detection of myocardial ischemia.


Assuntos
Doença da Artéria Coronariana , Estenose Coronária , Isquemia Miocárdica , Humanos , Constrição Patológica , Isquemia Miocárdica/diagnóstico , Eletrocardiografia , Estenose Coronária/diagnóstico , Teste de Esforço
7.
Int J Mol Sci ; 25(1)2023 Dec 21.
Artigo em Inglês | MEDLINE | ID: mdl-38203284

RESUMO

Cytoplasmic Dynein is a multiple-subunit macromolecular motor protein involved in the transport process of cells. The Dynein intermediate chain (DIC) is one of the subunits of Dynein-1. In our previous studies, we showed that Pt-DIC may play an important role in the nuclear deformation of spermiogenesis in Portunus trituberculatus. Lamin B is essential for maintaining nuclear structure and functions. Surprisingly, Pt-Lamin B was expressed not only in the perinucleus but also in the pro-acrosome during spermiogenesis in P. trituberculatus. Studies have also shown that Dynein-1 can mediate the transport of Lamin B in mammals. Thus, to study the relationship of Pt-DIC and Pt-Lamin B in the spermatogenesis of P. trituberculatus, we knocked down the Pt-DIC gene in P. trituberculatus by RNAi. The results showed that the distribution of Pt-DIC and Pt-Lamin B in spermiogenesis was abnormal, and the colocalization was weakened. Moreover, we verified the interaction of Pt-DIC and Pt-Lamin B via coimmunoprecipitation. Therefore, our results suggested that both Pt-DIC and Pt-Lamin B were involved in the spermatogenesis of P. trituberculatus, and one of the functions of Dynein-1 is to mediate the transport of Lamin B in the spermiogenesis of P. trituberculatus.


Assuntos
Lamina Tipo B , Espermatogênese , Masculino , Animais , Espermatogênese/genética , Acrossomo , Dineínas do Citoplasma , Dineínas/genética , Mamíferos
8.
Clin Endocrinol (Oxf) ; 90(5): 702-710, 2019 05.
Artigo em Inglês | MEDLINE | ID: mdl-30739355

RESUMO

OBJECTIVE: We examined the associations between maternal age at menarche and anthropometry and metabolism in adolescent offspring. METHODS: Anthropometric, metabolic and blood pressure data were obtained from 304 girls and 190 boys aged 11-16 years attending school in Hangzhou (China). Age at menarche for both mothers and daughters was self-reported. Fasting blood samples were obtained and all participants underwent clinical examinations. Obesity was defined as BMI ≥95th percentile for age and sex. RESULTS: Older maternal age at menarche was associated with older age of their daughters at menarche (r = 0.21; P < 0.001). Mother's age at menarche was not associated with anthropometry or metabolism of daughters. However, younger maternal age at menarche was associated with increased hip and waist circumferences, and BMI SDS of their sons. Boys whose mothers were ≤13 years at menarche had an adjusted relative risk of obesity 3-fold greater than sons of mothers with a later menarcheal onset (2.96; 95% CI 1.49, 5.87). Among daughters, every 1-year increase in their age at menarche was associated with a 0.34 SDS reduction in BMI. Increasing age at menarche was also associated with reduced waist and hip circumferences (-1.5 and -1.8 cm/y, respectively) and waist-to-height ratio (-0.008 per year). Girls in the youngest menarcheal age tertile (8.8-11.6 years) had diastolic blood pressure 2.2 mm Hg higher than other girls (P = 0.029). CONCLUSIONS: Younger maternal age at menarche is associated with increased obesity risk in their sons, but not daughters. However, girls who experience menarche earlier have a less favourable anthropometric profile.


Assuntos
Pressão Sanguínea , Índice de Massa Corporal , Menarca , Mães/estatística & dados numéricos , Obesidade Infantil/epidemiologia , Caracteres Sexuais , Circunferência da Cintura , Razão Cintura-Estatura , Adolescente , Fatores Etários , Pressão Sanguínea/fisiologia , Criança , China/epidemiologia , Feminino , Humanos , Masculino , Menarca/fisiologia , Risco , Circunferência da Cintura/fisiologia
9.
Int J Syst Evol Microbiol ; 69(11): 3512-3518, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31454307

RESUMO

Micrococcus aloeverae,Micrococcus endophyticus, Micrococcus luteus and Micrococcus yunnanensis are phenotypically and genotypically closely related, and together comprise the M. luteus group. In this study, the taxonomic relationships among Micrococcus aloeverae, M. luteus and M. yunnanensis were re-evaluated by using polyphasic approaches. The similarity values of the concatenated housekeeping gene (gyrB, recA and rpoB) sequences shared by the type strains of M. aloeverae, M. luteus and M. yunnanensis ranged from 98.3 to 99.4 %. The average nucleotide identity, average amino acid identity and digital DNA‒DNA hybridization values among these three taxa were greater (97.1‒98.1 %, 96.8‒98.1 % and 75.0‒83.5 %, respectively) than the thresholds for bacterial species delineation, indicating that they belong to the same species, whereas those for M. endophyticus were clearly lower than the thresholds. In addition, phenotypic and chemotaxonomic characterization results also support the synonymy of these three taxa. Therefore, we propose that M. aloeverae and M. yunnanensis should be reclassified as later heterotypic synonyms of M. luteus.


Assuntos
Micrococcus luteus/classificação , Micrococcus/classificação , Filogenia , Técnicas de Tipagem Bacteriana , Composição de Bases , DNA Bacteriano/genética , Ácidos Graxos/química , Genes Bacterianos , Hibridização de Ácido Nucleico , RNA Ribossômico 16S/genética , Análise de Sequência de DNA
10.
Int J Syst Evol Microbiol ; 69(5): 1484-1489, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30888315

RESUMO

A Gram-stain-positive, rod-shaped, non-motile, catalase-negative and facultative anaerobic strain, L88T, was isolated from suan-tsai, a traditional Taiwanese fermented mustard green. Comparative analyses of 16S rRNA, pheS and rpoA gene sequences demonstrated that strain L88Twas a member of the genus Lactobacillus. On the basis of 16S rRNA gene sequence similarity, the type strains of Lactobacillus acidifarinae (98.2 % similarity), Lactobacillus namurensis (98.1 %), Lactobacillus zymae (98.1 %) and Lactobacillus spicheri (96.8 %) were the closest neighbours to this novel strain. The average nucleotide identity, digital DNA‒DNA hybridization and average amino acid identity values between L88T and its closest relatives were lower than 80, 30 and 90 %, respectively. Phenotypic and genotypic test results demonstrated that strain L88T represents a novel species of the genus Lactobacillus, for which the name Lactobacillus suantsaii sp. nov., is proposed. The type strain is L88T (=BCRC 12945T=NBRC 113535T).


Assuntos
Alimentos Fermentados/microbiologia , Microbiologia de Alimentos , Lactobacillus/classificação , Mostardeira/microbiologia , Filogenia , Técnicas de Tipagem Bacteriana , Composição de Bases , DNA Bacteriano/genética , Lactobacillus/isolamento & purificação , Hibridização de Ácido Nucleico , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Taiwan
11.
Fish Shellfish Immunol ; 95: 227-235, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31654766

RESUMO

Myeloid differentiation factor 88 (MyD88) is a universal and essential adaptor protein required for the Toll-like receptors (TLRs) pathway activation in invertebrates as well as in vertebrates. Herein, we characterized a MyD88 (Pt-MyD88) cDNA sequence in the swimming crab (Portunus trituberculatus). The Pt-MyD88 ORF is predicted to encode 469 peptides with an N-terminal death domain and a typical C-terminal TIR domain. Real-Time quantitative PCR analysis showed that the Pt-MyD88 transcriptions were constitutively expressed in hemocytes, gill, intestine, heart and muscle in normal crab. The expressions of Pt-MyD88 would be down-regulated by V. alginolyticus or LPS challenge, and be up-regulated by WSSV infection in hemocytes. Intracellular localization showed Pt-MyD88 was distributed mainly in the cytoplasm when it was over-expressed in human cell HEK293T or in Drosophila Schneider 2 (S2). Functionally, over-expression of Pt-MyD88 could either activate the NF-κB in HEK293T cells or activate the promoters of Drosophila antimicrobial peptide genes (AMPs) in S2 cell. In primary cultured hemocytes of swimming crab, after Pt-MyD88 was knocked-down by specific long double strand RNA, the expression of anti-lipopolysaccharide factor1 (ALF1), hyastatin3, crustin1 and crustin3 have been significantly inhibited, while the expression of other AMPs is normal compared to non-specific dsRNA treated cells.


Assuntos
Braquiúros/genética , Braquiúros/imunologia , Fator 88 de Diferenciação Mieloide/genética , Fator 88 de Diferenciação Mieloide/imunologia , Transdução de Sinais , Sequência de Aminoácidos , Animais , Proteínas de Artrópodes/química , Proteínas de Artrópodes/genética , Proteínas de Artrópodes/imunologia , Sequência de Bases , Linhagem Celular , Regulação para Baixo/imunologia , Drosophila , Feminino , Células HEK293 , Hemócitos/imunologia , Humanos , Lipopolissacarídeos/fisiologia , Masculino , Modelos Animais , Fator 88 de Diferenciação Mieloide/química , Filogenia , Regulação para Cima/imunologia , Vibrio alginolyticus/fisiologia , Vírus da Síndrome da Mancha Branca 1/fisiologia
12.
Environ Toxicol ; 34(8): 891-901, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31157515

RESUMO

Nepenthes plants are regarded as a kind of Traditional Chinese Medicine for several diseases but its anticancer activity remain unclear. The subject of this study is to evaluate the antiproliferation effects on oral cancer cells by Nepenthes plants using ethyl acetate extract of Nepenthes adrianii x clipeata (EANA). Cell viability was detected using MTS assay. Its detailed mechanisms including cell cycle, apoptosis, oxidative stress, and DNA damage were explored by flow cytometry or western blotting. For 24 hours EANA treatment, five kinds of oral cancer cells (CAL 27, Ca9-22, OECM-1, HSC-3, and SCC9) show IC50 values of cell viability ranging from 8 to 17 µg/mL but the viability of normal oral cells (HGF-1) remains over 80%. Subsequently, CAL 27 and Ca9-22 cells with high sensitivity to EANA were chosen to investigate the detailed mechanism. EANA displays the time course and concentration effects for inducing apoptosis based on flow cytometry (subG1 and annexin V analyses) and western blotting [cleaved poly (ADP-ribose) polymerase (c-PARP)]. Oxidative stress and DNA damage were induced by EANA treatments in oral cancer cells through reactive oxygen species (ROS), mitochondrial membrane potential disruption, mitochondrial superoxide, and γH2AX. All these changes of EANA treatments in oral cancer cells were reverted by the ROS scavenger N-acetylcysteine pretreatment. Therefore, EANA induces preferential killing, apoptosis, and DNA damage against oral cancer cells through oxidative stress.


Assuntos
Antineoplásicos Fitogênicos/farmacologia , Neoplasias Bucais/tratamento farmacológico , Estresse Oxidativo , Traqueófitas , Acetatos , Antineoplásicos Fitogênicos/uso terapêutico , Apoptose , Ciclo Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Dano ao DNA , Humanos , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Mitocôndrias/metabolismo , Neoplasias Bucais/metabolismo , Fitoterapia , Extratos Vegetais/farmacologia , Poli(ADP-Ribose) Polimerases/metabolismo , Espécies Reativas de Oxigênio/metabolismo
13.
BMC Genomics ; 19(Suppl 10): 904, 2018 Dec 31.
Artigo em Inglês | MEDLINE | ID: mdl-30598081

RESUMO

BACKGROUND: Human gut microbiome has an essential role in human health and disease. Although the major dominant microbiota within individuals have been reported, the change of gut microbiome caused by external factors, such as antibiotic use and bowel cleansing, remains unclear. We conducted this study to investigate the change of gut microbiome in overweight male adults after bowel preparation, where none of the participants had been diagnosed with any systemic diseases. METHODS: A total of 20 overweight, male Taiwanese adults were recruited, and all participants were omnivorous. The participants provided fecal samples and blood samples at three time points: prior to bowel preparation, 7 days after colonoscopy, and 28 days after colonoscopy. The microbiota composition in fecal samples was analyzed using 16S ribosome RNA gene amplicon sequencing. RESULTS: Our results demonstrated that the relative abundance of the most dominant bacteria hardly changed from prior to bowel preparation to 28 days after colonoscopy. Using the ratio of Prevotella to the sum of Prevotella and Bacteroides in the fecal samples at baseline, the participants were separated into two groups. The fecal samples of the Type 1 group was Bacteroides-dominant, and that of the Type 2 group was Prevotella-dominant with a noticeable presence Bacteroides. Bulleidia appears more in the Type 1 fecal samples, while Akkermensia appears more in the Type 2 fecal samples. Of each type, the gut microbial diversity differed slightly among the three collection times. Additionally, the Type 2 fecal microbiota was temporarily susceptible to bowel cleansing. Predictive functional analysis of microbial community reveals that their activities for the mineral absorption metabolism and arachidonic acid metabolism differed significantly between the two types. Depending on their fecal type, the variance of triglycerides and C-reactive protein also differed between the two types of participants. CONCLUSIONS: Depending upon the fecal type, the microbial diversity and the predictive functional modules of microbial community differed significantly after bowel preparation. In addition, blood biochemical markers presented somewhat associated with fecal type. Therefore, our results might provide some insights as to how knowledge of the microbial community could be used to promote health through personalized clinical treatment.


Assuntos
Fezes/microbiologia , Microbioma Gastrointestinal , Sobrepeso/microbiologia , Adulto , Biodiversidade , Humanos , Masculino , Pessoa de Meia-Idade , Adulto Jovem
14.
Fish Shellfish Immunol ; 81: 338-342, 2018 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-30017932

RESUMO

The housekeeping gene encoding ß-actin appears to be the most widely-used internal reference for gene expression studies in experimental animals or their cell lines. However, the effectiveness of ß-actin to normalize mRNA levels expression in many crustacean species is still object of debate. To date, it is still unclear if ß-actin is suitable to be utilized as the internal reference in qualitative real-time gene expression study in crab species. To address this concern, we evaluated 5 candidate reference genes encoding ß-actin, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), cyclophilin A, elongation factor 1-α (EF1-α), and 18 S ribosomal RNA (18 S rRNA) in the swimming crabs (Portunus trituberculatus) models. Our data showed that the ß-actin gene expression varied significantly across individual swimming crab individuals in gills or hemocytes and the expression of 18 S rRNA, EF1-α, cyclophilin or GAPDH gene were relatively stable compared to that of ß-actin. Moreover, the expression stability of the reference genes among different tissues in normal crabs or after WSSV challenge was also tested by geNorm and NormFinder software. Among tissues, 18 S rRNA was most stably expressed in different tissues, followed by cyclophilin A and EF1-α, compared to ß-actin and GAPDH. Upon to viral simulation, GAPDH was found to be the most stable internal control gene in gills and cyclophilin A was ranked as the most stable gene in hemocytes.


Assuntos
Proteínas de Artrópodes/genética , Braquiúros/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa/normas , Actinas/genética , Animais , Braquiúros/virologia , Ciclofilina A/genética , Expressão Gênica , Variação Genética , Gliceraldeído-3-Fosfato Desidrogenases/genética , Masculino , Fator 1 de Elongação de Peptídeos/genética , RNA Mensageiro/genética , RNA Ribossômico 18S/genética , Padrões de Referência , Vírus da Síndrome da Mancha Branca 1
15.
Asian-Australas J Anim Sci ; 31(1): 106-115, 2018 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-28728397

RESUMO

OBJECTIVE: The goal of this study was to investigate the effects of dietary standard ileal digestible (SID) valine:lysine ratios on performance, intestinal morphology, amino acids of liver and muscle, plasma indices and mRNA expression of branched-chain amino acid (BCAA) metabolism enzymes. METHODS: A total of 144 crossbred pigs (Duroc×Landrace×Large White) weaned at 28±4 days of age (8.79±0.02 kg body weight) were randomly allotted to 1 of 4 diets formulated to provide SID valine:lysine ratios of 50%, 60%, 70%, or 80%. Each diet was fed to 6 pens of pigs with 6 pigs per pen (3 gilts and 3 barrows) for 28 days. RESULTS: Average daily gain increased quadratically (p<0.05), the villous height of the duodenum, jejunum and ileum increased linearly (p<0.05) as the SID valine:lysine ratio increased. The concentrations of plasma α-keto isovaleric and valine increased linearly (p<0.05), plasma aspartate, asparagine and cysteine decreased (p<0.05) as the SID valine:lysine ratio increased. An increase in SID lysine:valine levels increased mRNA expression levels of mitochondrial BCAA transaminase and branched-chain α-keto acid dehydrogenase in the longissimus dorsi muscle (p<0.05). CONCLUSION: Using a quadratic model, a SID valine:lysine ratio of 68% was shown to maximize the growth of weaned pigs which is slightly higher than the level recommended by the National Research Council [6].

16.
Zhongguo Dang Dai Er Ke Za Zhi ; 20(11): 925-929, 2018 Nov.
Artigo em Zh | MEDLINE | ID: mdl-30477624

RESUMO

Kallmann syndrome (KS) is a rare pediatric disease with major manifestations of olfactory dysfunction and hypogonadotropic hypogonadism. Five children (4 boys and 1 girl) with KS reported in this article were aged between 6 months and 19 years at the time when they attended the hospital. All the children had the clinical manifestation of hypogonadotropic hypogonadism; in addition, three children had olfactory dysfunction (two were found to have olfactory bulb dysplasia on magnetic resonance imaging), one had cleft lip and palate, and one had micropenis and cryptorchidism with right renal agenesis during infancy. All the five children had normal karyotype and their parents had normal clinical phenotypes. The uncle of one child had underdeveloped secondary sexual characteristics and olfactory disorder since childhood. High-throughput sequencing found two known heterozygous missense mutations in the FGFR1 gene, i.e., c.1097C>T(p.P366L) and c.809G>C(p.G270A), in two children. One child had a novel frameshift mutation, c.1877_1887/p.S627Tfs*6, in the KAL1 gene; this deletion mutation caused a frameshift in base sequence and produced truncated proteins, which led to a significant change in protein structure, and thus it was highly pathogenic. It is concluded that KS has great clinical and genetic heterogeneity and can be accompanied by incomplete dominant inheritance and that gene detection helps with the diagnosis of this disease.


Assuntos
Hipogonadismo , Síndrome de Kallmann , Adolescente , Criança , Pré-Escolar , Análise Mutacional de DNA , Proteínas da Matriz Extracelular , Feminino , Heterozigoto , Humanos , Lactente , Masculino , Mutação , Proteínas do Tecido Nervoso , Receptor Tipo 1 de Fator de Crescimento de Fibroblastos , Adulto Jovem
17.
Carcinogenesis ; 35(7): 1500-9, 2014 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-24445145

RESUMO

The mouse double minute 4 (MDM4) oncoprotein may inhibit tumorigenesis by regulating the apoptotic mediator p53. Ubiquitin-specific protease 2a (USP2a) is a deubiquitinating enzyme that protects MDM4 against degradation, so USP2-MDM4 interaction may be a key determinant of the malignant potential of human cancers. MDM4 and USP2a, as well as the MDM4-USP2a complex, were more highly expressed in glioblastoma multiforme tissue samples from patients with good prognosis compared with patients with poor prognosis. Analysis of the prognostic parameters indicated that MDM4 expression was positively correlated with an increased likelihood for survival. Compared with the poor prognosis patients, mitochondria from good prognosis glioma patients contained higher levels of both MDM4 and the proapoptotic protein p53Ser46(P). In U87MG glioma cell line, the overexpression of MDM4 enhanced ultraviolet (UV)-induced cytochrome c release and apoptosis. In contrast, MDM4 knockdown decreased mitochondrial p53Ser46(P) levels and rescued cells from UV-induced apoptosis. The expression of MDM4 and USP2a were positively correlated with each other. MDM4-USP2a complexes were found only in the cytoplasmic fraction, whereas the mitochondrial fraction contained MDM4-p53Ser46(P) and MDM4-Bcl-2 complexes. Overexpression of USP2a increased p53 and p53Ser46(P) levels in the mitochondria, whereas simultaneous MDM4 knockdown completely reversed this effect. UV-induced apoptosis was reduced by USP2a knockdown but restored by the simultaneous overexpression of MDM4. This apoptotic response was reduced by knockdown of p53 but not p21. Our results suggest that USP2a binds to and stabilizes MDM4; thus in turn, it enhances the mitochondrial localization of p53 and promotes apoptosis in glioma cells.


Assuntos
Apoptose , Endopeptidases/metabolismo , Glioblastoma/patologia , Proteínas Nucleares/metabolismo , Proteínas Proto-Oncogênicas/metabolismo , Transdução de Sinais , Proteína Supressora de Tumor p53/metabolismo , Animais , Western Blotting , Proteínas de Ciclo Celular , Núcleo Celular/metabolismo , Proliferação de Células , Inibidor de Quinase Dependente de Ciclina p21/genética , Inibidor de Quinase Dependente de Ciclina p21/metabolismo , Citoplasma/metabolismo , Endopeptidases/genética , Imunofluorescência , Glioblastoma/metabolismo , Glioblastoma/mortalidade , Humanos , Técnicas Imunoenzimáticas , Imunoprecipitação , Camundongos , Mitocôndrias/metabolismo , Mitocôndrias/patologia , Proteínas Nucleares/genética , Proteínas Proto-Oncogênicas/genética , RNA Mensageiro/genética , Reação em Cadeia da Polimerase em Tempo Real , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Células Tumorais Cultivadas , Proteína Supressora de Tumor p53/genética , Ubiquitina Tiolesterase
18.
Dev Genes Evol ; 224(3): 147-57, 2014 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-24869661

RESUMO

The gene doublesex (dsx) has shown deep conservation in the sex determination in many organisms. Environmental stimuli initiate a switch in the reproductive strategy of Daphnia pulex from asexual to sexual reproduction; however, occasionally, changes in environmental conditions will not lead to this transition. So study genetic responses to environmental stimuli and the molecular basis for the switch of reproductive stages are urgently needed. Therefore, we isolated and sequenced a D. pulex doublesex1 gene (Dpdsx1) and analyzed its expression and location by quantitative polymerase chain reaction (qPCR) and whole-mount in situ hybridization in D. pulex during different stages of reproduction. The predicted amino acid sequence has 335 amino acids that contained one DM domain and one dimerization domain, which is characteristic of insect orthologs of Dsx. Real-time PCR showed that Dpdsx1 expression decreased significantly (P < 0.05) in different reproductive stages in the following order: male, parthenogenetic female, ephippial female, resting egg, and juvenile female. Whole-mount in situ hybridization revealed that Dpdsx1 is expressed in the first antennae, first thoracic limb and compound eye in males, whereas expression levels in the corresponding sites of parthenogenetic and ephippial females were relatively weak. Dpdsx1 could not be detected in the gonads of males or ephippial and parthenogenetic females. Taken together, these different reproductive stages' and sex specific expression patterns are regulated temporally and spatially. We speculate that Dpdsx1 may involve in switching different stages of reproduction and in sexual differentiation in D. pulex.


Assuntos
Proteínas de Artrópodes/genética , Proteínas de Artrópodes/metabolismo , Proteínas de Ligação a DNA/genética , Proteínas de Ligação a DNA/metabolismo , Daphnia/genética , Sequência de Aminoácidos , Animais , Proteínas de Artrópodes/química , Clonagem Molecular , Proteínas de Ligação a DNA/química , Daphnia/fisiologia , Dados de Sequência Molecular , Técnicas de Amplificação de Ácido Nucleico , Filogenia , Reação em Cadeia da Polimerase em Tempo Real , Reprodução , Alinhamento de Sequência
19.
Cancer Cell Int ; 14: 34, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-24739220

RESUMO

Prostate cancer is a gland tumor in the male reproductive system. It is a multifaceted and genomically complex disease. Transmembrane protease, serine 2 and v-ets erythroblastosis virus E26 homolog (TMPRSS2-ERG) gene fusions are the common molecular signature of prostate cancer. Although tremendous advances have been made in unraveling various facets of TMPRSS2-ERG-positive prostate cancer, many research findings must be sequentially collected and re-interpreted. It is important to understand the activation or repression of target genes and proteins in response to various stimuli and the assembly in signal transduction in TMPRSS2-ERG fusion-positive prostate cancer cells. Accordingly, we divide this multi-component review ofprostate cancer cells into several segments: 1) The role of TMPRSS2-ERG fusion in genomic instability and methylated regulation in prostate cancer and normal cells; 2) Signal transduction cascades in TMPRSS2-ERG fusion-positive prostate cancer; 3) Overexpressed genes in TMPRSS2-ERG fusion-positive prostate cancer cells; 4) miRNA mediated regulation of the androgen receptor (AR) and its associated protein network; 5) Quantitative control of ERG in prostate cancer cells; 6) TMPRSS2-ERG encoded protein targeting; In conclusion, we provide a detailed understanding of TMPRSS2-ERG fusion related information in prostate cancer development to provide a rationale for exploring TMPRSS2-ERG fusion-mediated molecular network machinery.

20.
Prev Med ; 67: 35-40, 2014 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-24967955

RESUMO

OBJECTIVE: This study aimed to evaluate the diagnostic value of Waist-to-Height Ratio in early detection of obesity and metabolic syndrome in Chinese children and adolescents. METHOD: A cross-sectional study was conducted in six cities in China in 2010 with 16,914 children and adolescents aged 7-17 years. Participants were randomly divided into the training and testing sets. Diagnostic values were estimated using sensitivity, specificity and areas under receiver operating characteristic curves. RESULTS: The coefficients of variation of Waist-to-Height Ratio among age groups were lower than that of body mass index and waist circumstance. The area under receiver operating characteristic curve of Waist-to-Height Ratio was 0.968 in boys and 0.949 in girls for general obesity evaluation, and 0.983 in boys and 0.984 in girls for central obesity. The optimal cut-offs of Waist-to-Height Ratio were 0.47 in boys and 0.45 in girls in the training set and validated in the testing set. For metabolic syndrome evaluation, the sensitivity and specificity were 0.858 and 0.825 in boys, 0.864 and 0.812 in girls under the suggested cut-offs. CONCLUSION: Waist-to-Height Ratio was a simple, effective and practical tool for mass screening childhood obesity and metabolic syndrome in China. It will have potential values in public health practice.


Assuntos
Programas de Rastreamento/métodos , Síndrome Metabólica/diagnóstico , Obesidade Infantil/diagnóstico , Razão Cintura-Estatura , Adolescente , Criança , China , Estudos Transversais , Feminino , Humanos , Masculino , Obesidade Abdominal/diagnóstico , Curva ROC , Sensibilidade e Especificidade
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