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1.
J Immunol ; 210(3): 229-244, 2023 02 01.
Artigo em Inglês | MEDLINE | ID: mdl-36548476

RESUMO

The braking mechanisms to protect the host from tissue damage and inflammatory disease caused by an overexuberant immune response are common in many T cell subsets. However, the negative regulation of T cell responses and detailed mechanisms are not well understood in early vertebrates. In the current study, using a Nile tilapia (Oreochromis niloticus) model, we investigated the suppression of T cell immunity by IL-10. Tilapia encodes an evolutionarily conserved IL-10, whose expression in lymphocytes is markedly induced during the primary adaptive immune response against Aeromonas hydrophila infection. Activated T cells of tilapia produce IL-10, which in turn inhibits proinflammatory cytokine expression and suppresses PHA-induced T cell activation. Moreover, administration of IL-10 impairs the proliferation of tilapia T cells, reduces their potential to differentiate into Th subsets, and cripples the cytotoxic function, rendering the animals more vulnerable to pathogen attack. After binding to its receptor IL-10Ra, IL-10 activates the JAK1/STAT3 axis by phosphorylation and enhances the expression of the suppressor of cytokine signaling 3 (SOCS3), which in turn attenuates the activation of the NF-κB and MAPK/ERK signaling pathways, thus suppressing the T cell response of tilapia. Our findings elucidate a negative regulatory mechanism of T cell immunity in a fish species and support the notion that the braking mechanism of T cells executed through IL-10 existed prior to the divergence of the tetrapod lineage from teleosts. Therefore, this study, to our knowledge, provides a novel perspective on the evolution of the adaptive immune system.


Assuntos
Ciclídeos , Doenças dos Peixes , Tilápia , Animais , NF-kappa B/metabolismo , Tilápia/metabolismo , Interleucina-10/metabolismo , Proteínas Supressoras da Sinalização de Citocina/metabolismo , Proteínas de Peixes/metabolismo
2.
Chemistry ; 30(1): e202302147, 2024 Jan 02.
Artigo em Inglês | MEDLINE | ID: mdl-37584162

RESUMO

In recent years, the thermochromic hydrogel was acted as suitable sandwiching material to adjust light transmission. However, to accurately control the thermochromic temperature in a wide range still was a significant challenge. Here, a simple method was explored to prepare hydrogels with gradient opaque-transparent transition thermochromic temperature from 5 °C to 53 °C, which was regulated by the aggregation state of sodium dodecyl sulfate micelles by adding potassium tartrate hemihydrate and cations. Using Li+ , Na+ , and K+ as cations, the accuracy was controlled at 1 °C. Moreover, the transmittance of the hydrogel was not changed when the thermochromic temperature was adjusted. As a result, an intelligent window was fabricated by utilizing thermochromic hydrogel as a sandwiching layer into the outer glass layers, which could effectively and stably regulate the visible and infrared light. The temperature monitors/detectors were also designed, which showed excellent temperature monitoring/detecting ability. Therefore, this low-cost, high-efficient, large-scale prepared thermochromic hydrogel provided more potential for intelligent temperature devices.

3.
Langmuir ; 40(19): 9985-9992, 2024 May 14.
Artigo em Inglês | MEDLINE | ID: mdl-38699919

RESUMO

The limited specific surface area (SSA), long preparation period, and high cost are significant challenges for carbon xerogels (CXs). To overcome these limitations, we propose an approach to prepare tannin-resorcinol-formaldehyde-based CXs through template-catalyzed in situ polymerization. ZnCl2 acts as a catalyst and significantly accelerates the polymerization reaction through the coordination of Zn2+ to the carbonyl group in formaldehyde, while atmospheric drying instead of special drying and without solvent exchange reduces the preparation period to 24 h. In addition, ZnCl2 acts as an activator for the formation of many pores. Plant-derived tannins not only reduce the preparation cost but also regulate the pore structure. The resulted CXs with hierarchical porous structures show an optimal SSA of 1308 m2/g, high adsorption capabilities (for cationic, nitrosoaniline dyes, metal, and nonmetallic ions, especially for methylene blue with 454.93 mg/g), low shrinkage down to 10%, and reusability with 92.9% retention after 5 cycles. This work provides a promising and cost-effective method for the large-scale preparation of porous carbon materials with large SSA, offering potential applications in adsorption, energy storage, and catalysis.

4.
Cell Mol Life Sci ; 80(8): 219, 2023 Jul 20.
Artigo em Inglês | MEDLINE | ID: mdl-37470873

RESUMO

Recent advances highlight a key role of transient fasting in optimizing immunity of human and mouse. However, it remains unknown whether this strategy is independently acquired by mammals during evolution or instead represents gradually evolved functions common to vertebrates. Using a tilapia model, we report that T cells are the main executors of the response of the immune system to fasting and that dietary restriction bidirectionally modulates T cell immunity. Long-term fasting impaired T cell immunity by inducing intense autophagy, apoptosis, and aberrant inflammation. However, transient dietary restriction triggered moderate autophagy to optimize T cell response by maintaining homeostasis, alleviating inflammation and tissue damage, as well as enhancing T cell activation, proliferation and function. Furthermore, AMPK is the central hub linking fasting and autophagy-controlled T cell immunity in tilapia. Our findings demonstrate that dietary restriction to optimize immunity is an ancient strategy conserved in vertebrate evolution, providing novel perspectives for understanding the adaptive evolution of T cell response.


Assuntos
Linfócitos T , Tilápia , Animais , Humanos , Camundongos , Vertebrados/genética , Ativação Linfocitária , Autofagia/genética , Inflamação , Imunidade Adaptativa , Mamíferos
5.
Arch Gynecol Obstet ; 310(3): 1687-1695, 2024 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-38713294

RESUMO

PURPOSE: To compare differences in euploidy rates for blastocysts in preimplantation genetic testing for aneuploidy (PGT-A) cycles after gonadotropin-releasing hormone agonist (GnRH-a) long and short protocols, GnRH-antagonist (GnRH-ant) protocol, progestin-primed ovarian stimulation and mild stimulation protocols, and other ovary stimulation protocols. METHODS: This was a retrospective cohort study from the Assisted Reproductive Medicine Department of Shanghai First Maternity and Infant Hospital. A total of 1657 PGT-A cycles with intracytoplasmic sperm injection after different controlled ovary hyperstimulation protocols were analyzed, and a total of 3154 embryos were biopsied. Differences in euploidy rate per embryo biopsied, embryo euploidy rate per oocyte retrieved and cycle cancellation rate were compared. RESULTS: For the PGT-A cycles, the euploidy rate per embryo biopsied was lower in the GnRH-ant protocol than in the GnRH-a long protocol (53.26 vs. 58.68%, respectively). Multiple linear regression showed that the GnRH-ant protocol was associated with a lower euploidy rate per embryo biopsied (ß = -0.079, p = 0.011). The euploidy rate per embryo biopsied was not affected by total gonadotropin dosage, duration of stimulation and number of oocytes retrieved. The embryo euploidy rate per oocyte retrieved was similar in all protocols and was negatively correlated with the total number of oocytes retrieved (ß = -0.003, p = 0.003). CONCLUSION: Compared with the GnRH-a long protocol, the GnRH-ant protocol was associated with a lower euploidy rate per embryo biopsied. The total gonadotropin dosage, duration of stimulation and number of oocytes retrieved did not appear to significantly influence euploidy rates.


Assuntos
Blastocisto , Hormônio Liberador de Gonadotropina , Indução da Ovulação , Diagnóstico Pré-Implantação , Humanos , Feminino , Estudos Retrospectivos , Indução da Ovulação/métodos , Adulto , Hormônio Liberador de Gonadotropina/agonistas , Hormônio Liberador de Gonadotropina/antagonistas & inibidores , Injeções de Esperma Intracitoplásmicas , Gravidez , Aneuploidia
6.
Reprod Biol Endocrinol ; 20(1): 141, 2022 Sep 22.
Artigo em Inglês | MEDLINE | ID: mdl-36138458

RESUMO

BACKGROUND: Frozen-thawed embryo transfer (FET) is thought to be associated with obstetric and neonatal complications after in vitro fertilization/intracytoplasmic single sperm injection (IVF/ICSI) treatment. The study aimed to determine whether the endometrial preparation protocol is an influencing factor for these complications. METHODS: We conducted a retrospective cohort study of 3,458 women who had singleton deliveries after IVF/ICSI-FET treatment at the Centre for Reproductive Medicine of Shanghai First Maternity and Infant Hospital between July 2016 and April 2021. The women were divided into three groups according to the endometrial preparation protocols: 2,029 women with programmed cycles, 959 with natural cycles, and 470 with minimal ovarian stimulation cycles. The primary outcomes were the incidence rates of obstetric and neonatal complications, namely, hypertensive disorders of pregnancy (HDP), gestational diabetes mellitus (GDM), intrahepatic cholestasis of pregnancy (ICP), placenta previa, preterm rupture of membranes (PROM), preterm delivery, postpartum haemorrhage, large for gestational age (LGA), small for gestational age (SGA), and macrosomia. RESULTS: After adjustments for confounding variables by multivariate logistic regression analysis, the results showed that programmed cycles had an increased risk of HDP (aOR = 1.743; 95% CI, 1.110-2.735; P = 0.016) and LGA (aOR = 1.269; 95% CI, 1.011-1.592; P = 0.040) compared with natural cycles. Moreover, programmed cycles also increased the risk of LGA (aOR = 1.459; 95% CI, 1.083-1.965; P = 0.013) but reduced the risk of SGA (aOR = 0.529; 95% CI, 0.348-0.805; P = 0.003) compared with minimal ovarian stimulation cycles. There were no significant differences between natural cycles and minimal ovarian stimulation cycles. CONCLUSIONS: During IVF/ICSI-FET treatment, the risk of HDP and LGA was increased in women with programmed cycles. Therefore, for patients with thin endometrium, irregular menstruation or no spontaneous ovulation, minimal ovarian stimulation cycles may be a relatively safer option than programmed cycles.


Assuntos
Transferência Embrionária , Sêmen , China/epidemiologia , Criopreservação , Transferência Embrionária/efeitos adversos , Endométrio , Feminino , Fertilização in vitro/efeitos adversos , Humanos , Recém-Nascido , Masculino , Gravidez , Resultado da Gravidez/epidemiologia , Estudos Retrospectivos
7.
FASEB J ; 35(4): e21457, 2021 04.
Artigo em Inglês | MEDLINE | ID: mdl-33689192

RESUMO

As fish constitute the first evolutionary group with primordial T cells, they are of importance for understanding the origin and evolution of adaptive immunity. Yet, the knowledge about how ancestral T cells function remains limited. Therefore, the teleost model Nile tilapia (Oreochromis niloticus) was used in this study to investigate the regulatory mechanisms of T-cell immunity in fish. We identified an evolutionarily conserved canonical NF-κB signaling pathway in Nile tilapia, which participates in primary adaptive immune response during Streptococcus agalactiae infection. Blockade of NF-κB activity severely impairs T-cell activation and expansion, rendering the animals more vulnerable to pathogen attack. Meanwhile, NF-κB signaling is indispensable for fish T cells to produce IL-17A during the antibacterial immune response. Moreover, IL-17A binds its receptor IL-17RA, initiates the ACT1-TRAF6-TAK1 axis, and triggers NF-κB-dependent T-cell activation, thus forming a positive feedback loop of T-cell immunity in Nile tilapia. Furthermore, IL-17A seems to promote innate immunity by regulating pro-inflammatory cytokines via TRAF6-NF-κB axis, indicating the presence of an NF-κB-dependent IL-17A signaling pathway for coordinating adaptive and innate immunity in fish. Our results suggest that fish NF-κB couples TCR and IL-17 signals to modulate ancestral T-cell immunity against bacterial infection, and the regulation of T-cell immunity by NF-κB and IL-17 is a strategy that existed prior to the divergence of the tetrapod lineage from teleost fish. This study, therefore, provides a new perspective on the evolution of adaptive immunity.


Assuntos
Infecções Bacterianas/imunologia , Interleucina-17/metabolismo , NF-kappa B/metabolismo , Linfócitos T/imunologia , Animais , Ciclídeos/imunologia , Ciclídeos/metabolismo , Doenças dos Peixes/imunologia , Peixes , Imunidade Celular/imunologia , Ativação Linfocitária/imunologia , Receptores de Antígenos de Linfócitos T/metabolismo , Transdução de Sinais/imunologia
8.
J Pineal Res ; 72(1): e12778, 2022 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-34726796

RESUMO

Increasing evidence suggests that in vitro fertilization (IVF) may be associated with an increased risk of developing obesity and metabolic diseases later in life in the offspring. Notably, the addition of melatonin to culture medium may improve embryo development and prevent cardiovascular dysfunction in IVF adult mice. This study aimed to determine if melatonin supplementation in the culture medium can reverse impaired glucose metabolism in IVF mice offspring and the underlying mechanisms. Blastocysts used for transfer were generated by natural mating (control group) or IVF with or without melatonin (10-6  M) supplementation (mIVF and IVF group, respectively) in clinical-grade culture media. Here, we first report that IVF decreased hepatic expression of Fbxl7, which was associated with impaired glucose metabolism in mice offspring. Melatonin addition reversed the phenotype by up-regulating the expression of hepatic Fbxl7. In vitro experiments showed that Fbxl7 enhanced the insulin signaling pathway by degrading RhoA through ubiquitination and was up-regulated by transcription factor Foxa2. Specific knockout of Fbxl7 in the liver of adult mice, through tail intravenous injection of recombinant adeno-associated virus, impaired glucose tolerance, while overexpression of hepatic Fbxl7 significantly improved glucose tolerance in adult IVF mice. Thus, the data suggest that Fbxl7 plays an important role in maintaining glucose metabolism of mice, and melatonin supplementation in the culture medium may rescue the long-term risk of metabolic diseases in IVF offspring.


Assuntos
Melatonina , Animais , Blastocisto , Meios de Cultura , Suplementos Nutricionais , Fertilização in vitro , Glucose , Melatonina/farmacologia , Camundongos
9.
Fish Shellfish Immunol ; 127: 419-426, 2022 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-35779809

RESUMO

Interleukin-2 inducible T cell kinase (ITK) plays a predominant role in the T-cell receptor (TCR) signaling cascade to ensure valid T-cell activation and function. Nevertheless, whether it regulates T-cell response of early vertebrates remains unknown. Herein, we investigated the involvement of ITK in the lymphocyte-mediated adaptive immune response, and its regulation to T-cell activation in the Nile tilapia Oreochromis niloticus. Both sequence and structure of O. niloticus ITK (OnITK) were remarkably conserved with its homologues from other vertebrates, implying its potential conserved function. OnITK mRNA was extensively expressed in lymphoid-related tissues, and with the relative highest level in peripheral blood. Once Nile tilapia was infected by Edwardsiella piscicida, OnITK in splenic lymphocytes was significantly up-regulated on 7-day post infection at both transcription and translation levels, suggesting that OnITK might involve in the primary adaptive immune response of teleost. Furthermore, upon splenic lymphocytes were stimulated by T-cell specific mitogen PHA, OnITK mRNA and protein levels were dramatically elevated. More importantly, treatment of splenic lymphocytes with specific inhibitor significantly crippled OnITK expression, which in turn impaired the inducible expression of T-cell activation markers IFN-γ, IL-2 and CD122, indicating the critical roles of ITK in regulating T-cell activation of Nile tilapia. Taken together, our results suggest that ITK takes part in the lymphocyte-mediated adaptive immunity of tilapia, and is indispensable for T-cell activation of teleost. Our findings thus provide novel evidences for understanding the mechanism regulating T-cell immunity of early vertebrates, as well as the evolution of adaptive immune system.


Assuntos
Ciclídeos , Animais , Proteínas de Peixes/química , Interleucina-2/genética , Ativação Linfocitária/genética , Proteínas Tirosina Quinases , RNA Mensageiro/metabolismo , Receptores de Antígenos de Linfócitos T/genética , Receptores de Antígenos de Linfócitos T/metabolismo , Linfócitos T
10.
Fish Shellfish Immunol ; 128: 216-227, 2022 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-35934242

RESUMO

As a pleiotropic cytokine mainly secreted by CD4+ T cells, interleukin (IL)-22 plays an important role in immune regulation and infection elimination. Despite IL-22 homologues have been identified in non-mammal, whether and how IL-22 participates in the adaptive immune response of early vertebrates have not been fully addressed. In this study, we identified an evolutionarily conserved IL-22 from Nile tilapia Oreochromis niloticus (defined as OnIL-22), proved by its properties regarding sequence, gene structure, functional domain, tertiary structure and phylogeny. IL-22 was broadly expressed in lymphoid-related tissues of tilapia, and with relatively higher levels in skin, gill, intestine and liver. The expression of OnIL-22 in spleen lymphocytes was markedly induced at the adaptive immune stage after Streptococcus agalactiae infection. Moreover, once lymphocytes were activated by PMA plus ionomycin or T-cell specific mitogen PHA in vitro, OnIL-22 expression was obviously up-regulated at both mRNA and protein levels. These results thus suggest that activated T cells produce IL-22 to take part in the adaptive immune response of tilapia. Furthermore, treatment of lymphocytes with recombinant OnIL-22 increased the expression of genes related to proliferation and survival, and further promoted the proliferation and reduced the apoptosis of lymphocytes during bacterial infection or T-cell activation. These cellular effects of IL-22 seem to be associated with JAK1/STAT3 axis downstream of IL-22, because IL-22 application not only elevated the mRNA expression of JAK1 and STAT3, but also enhanced their phosphorylation in lymphocytes. Altogether, we suggest that activated T cells produce IL-22 to promote lymphocyte proliferation and survival probability via JAK1/STAT3 signaling pathway, thus participating in adaptive immune response of Nile tilapia. Our study therefore provides helpful perspective for understanding the function and mechanism of adaptive immune system in teleost.


Assuntos
Ciclídeos , Doenças dos Peixes/imunologia , Proteínas de Peixes/metabolismo , Interleucinas/metabolismo , Infecções Estreptocócicas , Animais , Proliferação de Células , Citocinas/genética , Regulação da Expressão Gênica , Ionomicina , Mitógenos , RNA Mensageiro/metabolismo , Infecções Estreptocócicas/veterinária , Streptococcus agalactiae/fisiologia , Linfócitos T , Interleucina 22
11.
Arch Gynecol Obstet ; 306(5): 1607-1615, 2022 11.
Artigo em Inglês | MEDLINE | ID: mdl-35904610

RESUMO

AIMS: The various diagnostic criteria for polycystic ovary syndrome (PCOS) raised problem for PCOS research worldwide. PCOS has been demonstrated to be significantly associated with immune response. We aimed to identify several immune-related biomarkers and construct a nomogram model for diagnosis in PCOS. METHODS: The mRNA expression data were downloaded from Gene Expression Omnibus (GEO) database. Significant immune-related genes were identified to be the biomarkers for the diagnosis of PCOS using random forest model (RF), support vector machine model (SVM) and generalized linear model (GLM). The key biomarkers were selected from the optimal model and were utilized to construct a diagnostic nomogram. Receiver operating characteristic (ROC) curves was used to evaluate diagnostic ability of nomogram. Moreover, the relative proportion of 22 immune cell types was calculated by CIBERSORT algorithm. RESULTS: Four immune-related biomarkers (cAMP, S100A9, TLR8 and IL6R) were demonstrated to be highly expressed in PCOS. The nomogram constructed on the ground of the four key biomarkers showed perfect performance in diagnosis of PCOS, whose AUC were greater than 0.7. Higher infiltrating abundance of neutrophils, resting NK cells and activated dendritic cells were observed in PCOS and were tightly associated with the four key biomarkers. CONCLUSIONS: This study identified several immune-related diagnostic biomarkers for PCOS patients. The diagnostic nomogram constructed based the biomarkers provide a theory foundation for clinical application. Multiple immune cells were associated with the expression of these four biomarkers and might played a vital role in the procession of PCOS.


Assuntos
Síndrome do Ovário Policístico , Biomarcadores , Feminino , Humanos , Síndrome do Ovário Policístico/diagnóstico , Síndrome do Ovário Policístico/genética , RNA Mensageiro , Curva ROC , Receptor 8 Toll-Like
12.
Fish Shellfish Immunol ; 113: 51-60, 2021 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-33798718

RESUMO

Killer cell lectin-like receptor G subfamily 1 (KLRG1) is a receptor generally expressed on effector CD8+ T cells or NK cells at terminal differentiation stage, and it will be highly induced for lymphocyte cytotoxicity upon pathogen infection or lymphocyte activation. However, little is known about the character or function of KLRG1 in lower vertebrates. In present study, we reappraised a molecule that previously defined as KLRG1 in the genomic sequence of Nile tilapia Oreochromis niloticus, and identified it as an atypical KLRG1-like molecule (defined as On-KLRG1-L), and illustrated its potential function serving as a marker representing effector T lymphocytes of fish species. On-KLRG1-L consists of two C-type lectin-like domains (CTLDs) without transmembrane region, and the tertiary structure of the CTLD is highly alike to that in mouse KLRG1. As a CTLD-containing protein, the recombinant On-KLRG1-L could bind PGN and several microbes in vitro. On-KLRG1-L was widely expressed in immune-associated tissues, with the highest expression level in the gill. Once Nile tilapia is infected by Aeromonas hydrophila, mRNA level of On-KLRG1-L in spleen lymphocytes were significantly up-regulated on 5 days after infection. Meanwhile, On-KLRG1-L protein was also induced on 5 or 8 days after A. hydrophila infection. Furthermore, we found both mRNA and protein levels of On-KLRG1-L were dramatically enhanced within several hours after spleen lymphocytes were activated by T cell-specific mitogen PHA in vitro. More importantly, the ratio of On-KLRG1-L+ T cells was also augmented after PHA stimulation. The observations suggested that the KLRG1-like molecule from Nile tilapia participated in lymphocyte activation and anti-bacterial adaptive immune response, and could serve as an activation marker of T lymphocytes. Our study thus provided new evidences to understand lymphocyte-mediated adaptive immunity of teleost.


Assuntos
Imunidade Adaptativa/genética , Ciclídeos/imunologia , Doenças dos Peixes/imunologia , Lectinas Tipo C/imunologia , Ativação Linfocitária/imunologia , Receptores Imunológicos/imunologia , Aeromonas hydrophila/fisiologia , Sequência de Aminoácidos , Animais , Biomarcadores/metabolismo , Doenças dos Peixes/microbiologia , Proteínas de Peixes/genética , Proteínas de Peixes/imunologia , Infecções por Bactérias Gram-Negativas/imunologia , Infecções por Bactérias Gram-Negativas/microbiologia , Infecções por Bactérias Gram-Negativas/veterinária , Lectinas Tipo C/genética , Estrutura Terciária de Proteína , Receptores Imunológicos/genética , Alinhamento de Sequência/veterinária
13.
Fish Shellfish Immunol ; 96: 13-25, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31760167

RESUMO

Tumor necrosis factor (TNF) receptor-associated factors (TRAFs) play crucial roles as signaling mediators for the TNF receptor (TNFR) superfamily and the interleukin-1 receptor/Toll-like receptor (IL-1R/TLR) superfamily. TRAFs collectively play important roles in multiple biological processes and organismal immunity. However, systematic identification of the TRAF gene family in teleost fish has not yet been reported, and there is little available information about its roles in innate immunity in Chinese tongue sole (Cynoglossus semilaevis), an aquaculture fish of high economic value. In the present study, we identified and characterized seven TRAF genes, namely, CsTRAF2a, CsTRAF2b, CsTRAF3, CsTRAF4, CsTRAF5, CsTRAF6 and CsTRAF7, in Chinese tongue sole, and the complete ORFs of the CsTRAFs were cloned. Sequence analysis revealed various genomic structures of the CsTRAFs and showed that they contain typical conserved domains compared with mammalian TRAFs. Phylogenetic analysis indicated the evolutionary relationships of TRAF family members in teleost fish and revealed an absence of TRAF1 in most species and TRAF5 in some species of teleosts. Analysis of the gene structures and motifs showed the diversity and distribution of exon-intron structures and conserved motifs in Chinese tongue sole and several other teleost species. Real-time quantitative PCR was used to investigate the expression patterns of CsTRAF genes in tissues of healthy fish and in the gills, livers and spleens of fish after bacterial infection with Vibrio harveyi. The results indicate that only CsTRAF2a is relatively highly expressed in the brain and that the other CsTRAFs are highly expressed in immune-related tissues and may participate in the immune response after infection with pathogenic bacteria. Functional analysis of CsTRAF3, CsTRAF4 and CsTRAF6 revealed that only CsTRAF6 could strongly activate the NF-кB pathway after overexpression of CsTRAF3, CsTRAF4 and CsTRAF6 in HEK-293T cells. This systematic analysis provided valuable information about the diverse roles of TRAFs in the innate immune response to pathogenic bacterial infection in teleost fish and will contribute to the functional characterization of CsTRAF genes in further research.


Assuntos
Doenças dos Peixes/imunologia , Linguados/genética , Linguados/imunologia , Regulação da Expressão Gênica/imunologia , Expressão Gênica/imunologia , Imunidade Inata/genética , Peptídeos e Proteínas Associados a Receptores de Fatores de Necrose Tumoral/genética , Animais , Proteínas de Peixes/genética , Proteínas de Peixes/metabolismo , Perfilação da Expressão Gênica/veterinária , Genoma , Família Multigênica/imunologia , Peptídeos e Proteínas Associados a Receptores de Fatores de Necrose Tumoral/metabolismo , Vibrio/fisiologia , Vibrioses/imunologia , Vibrioses/veterinária
14.
Fish Shellfish Immunol ; 106: 1120-1130, 2020 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-32971270

RESUMO

Ribosomal protein S6 kinase beta-1 (S6K1) is a serine/threonine kinase downstream of the mechanistic target of rapamycin (mTOR) pathway, and plays crucial roles in immune regulation. Although remarkable progress has been achieved with a mouse model, how S6K1 regulates adaptive immunity is largely unknown in early vertebrates. In this study, we identified an S6K1 from Nile tilapia Oreochromis niloticus (OnS6K1), and further investigated its potential regulatory role on the adaptive immunity of this fish species. Both sequence and structure of OnS6K1 were highly conserved with its homologs from other vertebrates and invertebrates. OnS6K1 was widely expressed in immune tissues, and with a relative higher expression level in the liver, spleen and head kidney. At the adaptive immune stage of Nile tilapia that infected with Aeromonas hydrophila, mRNA expression of OnS6K1 and its downstream effector S6 was significantly up-regulated in spleen lymphocytes. Meanwhile, their phosphorylation level was also enhanced during this process, suggesting that S6K1/S6 axis participated in the primary response of anti-bacterial adaptive immunity in Nile tilapia. Furthermore, after spleen lymphocytes were activated by the T cell-specific mitogen PHA or lymphocytes agonist PMA in vitro, mRNA and phosphorylation levels of S6K1 were elevated, and phosphorylation of S6 was also enhanced. Once S6K1 activity was blocked by a specific inhibitor, both mRNA and phosphorylation levels of S6 were severely impaired. More importantly, blockade of S6K1/S6 axis reduced the expression of T cell activation marker IFN-γ and CD122 in PHA-activated spleen lymphocytes, indicating the essential role of S6K1/S6 axis in regulating T cell activation of Nile tilapia. Together, our study suggests that S6K1 and its effector S6 regulate lymphocyte activation of Nile tilapia, and in turn promote lymphocyte-mediated adaptive immunity. This study enriched the mechanism of adaptive immune response in teleost and provided useful clues to understand the evolution of adaptive immune system.


Assuntos
Aeromonas hydrophila , Ciclídeos/imunologia , Doenças dos Peixes/imunologia , Proteínas de Peixes/imunologia , Infecções por Bactérias Gram-Negativas/imunologia , Proteínas Quinases S6 Ribossômicas 70-kDa/imunologia , Imunidade Adaptativa , Animais , Ciclídeos/genética , Proteínas de Peixes/genética , Infecções por Bactérias Gram-Negativas/veterinária , Ativação Linfocitária , Proteínas Quinases S6 Ribossômicas 70-kDa/genética , Linfócitos T/imunologia
15.
J Assist Reprod Genet ; 37(11): 2713-2722, 2020 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-32949002

RESUMO

PURPOSE: Tubulin beta eight class VIII (TUBB8) is essential for oogenesis, fertilization, and pre-implantation embryo development in human. Although TUBB8 mutations were recently discovered in meiosis-arrested oocytes of infertile females, there is no effective therapy for this gene mutation caused infertility. Our study aims to further reveal the infertility-causing gene mutations in the patient's family and to explore whether the infertility could be rescued by optimizing the conditions of embryo culture and finally achieve the purpose of making the patient pregnant. METHODS: Whole-exome sequence analysis and Sanger sequencing were performed on patients' family members to screen and identify candidate mutant genes. Construction of plasmids, in vitro transcription, microinjection of disease-causing gene cRNA, and immunofluorescence staining were used to recapitulate the infertility phenotype observed in patients and to understand the pathogenic principles. Simultaneously, overexpression of mutant and wild-type cRNA of the candidate gene in mouse oocytes at either germinal vesicle (GV) or metaphase II (MII) stage was performed in the rescue experiment. RESULTS: We first identified a novel heritable TUBB8 mutation (c.1041C>A: p.N347K) in the coding region which specifically affects the first mitosis and causes the developmental arrest of early embryos in a three-generation family. We further demonstrated that TUBB8 mutation could lead to abnormal spindle assemble. And moreover, additional expression of wild-type TUBB8 cRNA in the mouse oocytes in which the mutant TUBB8 were expressed can successfully rescue the developmental defects of resulting embryo and produce full-term offspring. CONCLUSIONS: Our study not only defines a novel mutation of TUBB8 causing the early cleavage arrest of embryos, but also provides an important basis for treating such female infertility in the future.


Assuntos
Infertilidade Feminina/genética , Oogênese/genética , Tubulina (Proteína)/genética , Animais , Divisão Celular/genética , Embrião de Mamíferos , Feminino , Humanos , Infertilidade Feminina/patologia , Masculino , Camundongos , Mitose/genética , Mutação/genética
16.
Fish Physiol Biochem ; 46(3): 881-890, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31909442

RESUMO

The Chinese tongue sole (Cynoglossus semilaevis) is a flatfish with distinctive asymmetry in its body coloration. The melanism (hyperpigmentation) in both the blind side and ocular side of C. semilaevis gives it an extremely low commercial value. However, the fundamental molecular mechanism of this melanism remains unclear. Melanocortin 1 receptor (MC1R), a GTP-binding protein-coupled receptor, is considered to play a vital role in the physiology of the vertebrate pigment system. In order to confirm the contribution of MC1R to the body coloration of C. semilaevis, the expression levels of Mc1r mRNA were measured in seven tissue types at different developmental stages of normal and melanistic C. semilaevis. The expression levels of Mc1r mRNA in the heart, brain, liver, kidney, ocular-side skin, and blind-side skin of melanistic C. semilaevis were significantly higher than that of normal C. semilaevis in all developmental stages. Moreover, the knocking down of Mc1r in the C. semilaevis liver cell line (HTLC) increased the expression of the downstream genes microphthalmia transcription factor (Mitf) and tyrosinase-related protein 1 (Tyrp1) in the pigmentation pathway. Thus, the present data suggest that MC1R might play important roles in Tyrp1- and Mitf-mediated pigment synthesis in C. semilaevis.


Assuntos
Proteínas de Peixes/genética , Linguados/genética , Receptor Tipo 1 de Melanocortina/genética , Animais , Encéfalo/metabolismo , Linhagem Celular , Rim/metabolismo , Fígado/metabolismo , Melaninas , Glicoproteínas de Membrana/genética , Fator de Transcrição Associado à Microftalmia/genética , Músculos/metabolismo , Miocárdio/metabolismo , Oxirredutases/genética , Pigmentação , Pele/metabolismo
17.
BMC Womens Health ; 19(1): 61, 2019 05 06.
Artigo em Inglês | MEDLINE | ID: mdl-31060530

RESUMO

BACKGROUND: This study aimed to evaluate the management of asymptomatic intrauterine lesions detected by ultrasonography. METHODS: Patients who underwent diagnostic hysteroscopy for asymptomatic lesions, including pre- and post-menopausal endometrial polyps, post-menopausal endometrial thickening (ET ≥5 mm) and reduplicative endometrial heterogeneity detected by transvaginal ultrasonography (TVUS), were recruited for this study. RESULTS: In the 792 recruited patients, the symptom-free focal masses within the uterine cavity detected by TVUS included 558 patients with pre- or post-menopausal endometrial polyps and 234 patients with postmenopausal endometrial thickening. No pre-menopausal patient presented with carcinoma. The polyp diameter (PD) was not identified as an independent risk factor for malignancy in this study. A significant difference (P = 0.036, < 0.05) in both benign and malignant endometrial lesions was observed between two groups of post-menopausal women stratified using an endometrial thickness cut-off of ≥11 mm. The TVUS was highly sensitive (94%) for pre-menopausal polyps. This technique had a specificity and positive predictive value of 84.4 and 92.7%, respectively, for postmenopausal polyps. The TVUS was clearly valuable for ruling out polyps, as indicated by a negative likelihood ratio (LR-) of 0.087. Among postmenopausal women with endometrial thickening, the area under the receiver operating characteristic curve was 0.828 (P < 0.001). An ET cut-off value of 12.5 mm yielded a sensitivity of 72.7% and specificity of 86%. CONCLUSION: We recommend follow-up alone for women with asymptomatic uterine polyps, particularly those who are pre-menopausal. Additionally, gynaecologists should consider risk factors such as age, obesity, polycystic ovarian syndrome, and diabetes. Prospective long-term follow-up studies should be conducted after hysteroscopic polypectomy to evaluate the recurrence rate of endometrial lesions.


Assuntos
Endométrio/cirurgia , Pós-Menopausa , Doenças Uterinas/cirurgia , Neoplasias Uterinas/cirurgia , Idoso , Endométrio/diagnóstico por imagem , Endométrio/patologia , Feminino , Humanos , Histeroscopia/métodos , Pessoa de Meia-Idade , Recidiva Local de Neoplasia/patologia , Pólipos/patologia , Estudos Retrospectivos , Ultrassonografia/métodos , Doenças Uterinas/diagnóstico por imagem , Doenças Uterinas/patologia , Neoplasias Uterinas/diagnóstico por imagem , Neoplasias Uterinas/patologia
18.
Chin Med Sci J ; 34(3): 221-225, 2019 Sep 30.
Artigo em Inglês | MEDLINE | ID: mdl-31601305

RESUMO

Here we report a case of heterotopic cornual pregnancy after in vitro fertilization who was diagnosed at 6 weeks after frozen embryos transfer. The heterotopic pregnancy was successfully terminated by transvaginal ultrasound-guided selective fetal reduction. At 38+1 weeks, she underwent a cesarean section and delivered a healthy 3300 g male infant with Apgar score of 10-10' evaluated at 1 min and 5 min.


Assuntos
Cesárea , Gravidez Cornual , Adulto , Feminino , Fertilização in vitro , Humanos , Gravidez , Gravidez Cornual/diagnóstico por imagem , Gravidez Cornual/terapia , Ultrassonografia
19.
Chemistry ; 24(56): 15119-15125, 2018 Oct 09.
Artigo em Inglês | MEDLINE | ID: mdl-30085376

RESUMO

Nucleobase pairs of adenine and uracil (A-U) from ribonucleic acid are of particular interest for various promising material properties. Herein, a novel polyacrylamide hydrogel with adhesive properties that are assisted by adenine and uracil has been designed and investigated. The incorporation of adenine and uracil enables the formation of a polyacrylamide hydrogel with remarkable adhesive behaviour with various materials including polytetrafluoroethylene (PTFE), plastics, rubber, glasses, metal, ceramics and wood. Moreover, the adhesive hydrogel can easily and directly adhere to humid biological tissues without any extra process, including heart, liver, spleen, lung, kidney, bone and muscle of mouse. More impressively, even after repeated peeling tests (10×), the AU-mediated polyacrylamide hydrogel still exhibits excellent durable adhesion for various materials. From mechanical contact tests, the adhesion energy of the A-U adhesive hydrogel is 47.9 J m-2 , which is nearly nine times that of polyacrylamide hydrogel (5.3 J m-2 ). The 90° peeling strength for aluminium, titanium, silica rubbers, glasses, PTFE and hogskin is 518, 645, 445, 396, 349, and 119 N m-1 , respectively. The multipurpose and durable adhesive behaviour of hydrogels assisted by adenine and uracil indicated the promise of nucleobase pairs from ribonucleic acid for the future development of adhesive materials.

20.
Fish Shellfish Immunol ; 77: 392-401, 2018 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-29635065

RESUMO

In mammals, microtubule-dependent trafficking could participate the immune response, where the motor proteins are suggested to play an important role in this process, while the related study in fish was rare. In this study, dctn5, a subunit of dyactin complex for docking motor protein, was obtained by previous immune QTL screening. The full-length cDNAs of two dctn5 transcript variants were cloned and identified (named dctn5_tv1 and dctn5_tv2, respectively). Tissue distribution showed that dctn5_tv1 was widely distributed and high transcription was observed in immune tissue (skin), while dctn5_tv2 was predominantly detected in gonad and very low in other tissues. Time-course expression analysis revealed that dctn5_tv1 could be up-regulated in gill, intestine, skin, spleen, and kidney after Vibrio harveyi challenge. Moreover, recombinant Dctn5_tv1 exhibited high antimicrobial activity against Escherichia coli and Streptococcus agalactiae due to binding to bacteria cells. Taken together, these data suggest Dctn5_tv1 is involved in immune response of bacterial invasion in Chinese tongue sole.


Assuntos
Complexo Dinactina/genética , Complexo Dinactina/imunologia , Doenças dos Peixes/imunologia , Linguados/genética , Linguados/imunologia , Regulação da Expressão Gênica/imunologia , Imunidade Inata/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Complexo Dinactina/química , Escherichia coli/fisiologia , Infecções por Escherichia coli/imunologia , Proteínas de Peixes/química , Proteínas de Peixes/genética , Proteínas de Peixes/imunologia , Perfilação da Expressão Gênica/veterinária , Filogenia , Alinhamento de Sequência/veterinária , Infecções Estreptocócicas/imunologia , Streptococcus agalactiae/fisiologia , Vibrio/fisiologia , Vibrioses/imunologia
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