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BACKGROUND/AIM: Pulp mineralisation is a survival process that may occur in the pulp of immature teeth following trauma. However, the mechanism of this process remains unclear. The aim of this study was to evaluate the histological manifestations of pulp mineralisation after intrusion in immature molars of rats. MATERIALS AND METHODS: Three-week-old male Sprague-Dawley rats were subjected to intrusive luxation of the right maxillary second molar by an impact force from a striking instrument through a metal force transfer rod. The left maxillary second molar of each rat was used as a control. The control and injured maxillae were collected at 3, 7, 10, 14, and 30 days after trauma (n = 15 per time group) and evaluated using haematoxylin and eosin staining and immunohistochemistry. Independent two-tailed Student's t-test was used for statistical comparison of the immunoreactive area. RESULTS: Pulp atrophy and mineralisation were observed in 30%-40% of the animals, and no pulp necrosis occurred. Ten days after trauma, pulp mineralisation, with osteoid tissue rather than reparative dentin, formed around the newly vascularised areas in the coronal pulp. CD90-immunoreactive cells were observed in the sub-odontoblastic multicellular layer in control molars, whereas the number of these cells was decreased in the traumatised teeth. CD105 localised in cells around the pulp osteoid tissue of the traumatised teeth, whereas in control teeth, it was only expressed in the vascular endothelial cells of capillaries in the odontoblastic or sub-odontoblastic layers. In specimens with pulp atrophy at 3-10 days after trauma, hypoxia inducible factor expression and CD11b-immunoreactive inflammatory cells increased. CONCLUSIONS: Following intrusive luxation of immature teeth without crown fractures in rats, no pulp necrosis occurred. Instead, pulp atrophy and osteogenesis around neovascularisation with activated CD105-immunoreactive cells were observed in the coronal pulp microenvironment characterised by hypoxia and inflammation.
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Polpa Dentária , Células Endoteliais , Masculino , Ratos , Animais , Ratos Sprague-Dawley , Necrose da Polpa Dentária , Dente MolarRESUMO
The shrub encroachment caused by Caragana species (mainly C. microphylla, C. korshinskii, C. tibetica, C. stenophylla, and C. pygmaea) in the north temperate zone has significant impacts on ecosystems. Understanding the distribution of Caragana species' responses to climate change is increasingly relevant to the dynamic of shrub encroachment. In this study, we gathered 1124 geographical distribution records for 5 Caragana species. Through principal component analysis and Pearson correlation analysis, 11 environmental variables were identified. We employed the maximum entropy (MaxEnt) model and utilized the current and future climate dataset from 2041 to 2060 based on two extreme climate scenarios (RCP2.6 and RCP8.5) and atmospheric circulation models (BCC_CSM1.1 and IPSLCM5A-LR) to assess the potential distribution patterns and dynamic change with global warming. The results showed the following: (1) Currently, the five Caragana species are mainly distributed in the central and western parts of the Inner Mongolia Autonomous Region, Mongolia, and the southern parts of Russia. (2) In the future, the habitable zone of C. microphylla and C. korshinskii will expand gradually, while the distribution probability of C. stenophylla, C. tibetica, and C. pygmaea will shrink significantly in 60-80% of the area, and the habitable area will fluctuate sharply. (3) The range of the five species of Caragana expansion area is projected to be 1229.43×106 km2-1412.32×106 km2, with the suitable habitats expected to extend northward in the future, primarily concentrated in central Mongolia and around Lake Baikal in Russia. This research provides guidance for protecting grassland resources and ensuring sustainable development under shrub encroachment.
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Caragana , Ecossistema , Monitoramento Ambiental , Simulação por Computador , ChinaRESUMO
Chrysanthemums (Chrysanthemum morifolium Ramat.) are ornamental flowers, which are famous worldwide. The mode of inheritance has great implications for the genetic analysis of polyploid species. However, genetic analysis of chrysanthemum has been hampered because of its controversial inheritance mode (disomic or hexasomic). To classify the inheritance mode of chrysanthemums, an analysis of three approaches was carried out in an F1 progeny of 192 offspring using 223 expressed sequence tag-simple sequence repeat (EST-SSR) markers. The analysis included segregation analysis, the ratio of simplex marker alleles linked in coupling to repulsion, as well as the transmission and segregation patterns of EST-SSR marker alleles. After segregation analysis, 204 marker alleles fit hexasomic inheritance and 150 marker alleles fit disomic inheritance, showing that marker alleles were inherited predominantly in a hexasomic manner. Furthermore, the results of the analysis of allele configuration and segregation behavior of five EST-SSR markers also suggested random pairing of chromosomes. Additionally, the ratio of simplex marker alleles linked in coupling to repulsion was 1:0, further supporting hexasomic inheritance. Therefore, it could be inferred that chrysanthemum is a complete or near-complete hexasome.
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Chrysanthemum , Alelos , Chrysanthemum/genética , Etiquetas de Sequências Expressas , Humanos , Repetições de Microssatélites , PoliploidiaRESUMO
The circadian clock can entrain to forced light-dark cycles by adjusting the phases and periods of flower opening and closing in ephemeral flowers. The responses of circadian rhythms to the same light conditions differ from species. However, the differences in internal genetic mechanisms underlying the different responses between species remain unclear. Iris domestica and I. dichotoma have ephemeral flowers and significantly divergent flower opening and closing times. The effects of different photoperiods (continuous darkness, 4L20D, 8L16D, 12L12D, 16L8D, 20L4D and continuous white light) on flower opening and closing, and expression patterns of seven genes (CRYPTOCHROME 1, PHYTOCHROME B, LATE ELONGATED HYPOCOTYL, PSEUDO RESPONSE REGULATOR 95, PHYTOCHROME INTERACTING FACTOR 4-like, SMUX AUXIN UP RNA 64-like and senescence-associated gene 39-like) involved in the circadian regulation of flower opening and closing were compared between I. domestica and I. dichotoma. Flower opening and closing in the two species exhibited circadian rhythms under continuous darkness (DD), but showed arrhythmia in continuous white light (LL). In the two species, keeping robust rhythms, strong synchronicity, rapid progressions of flower opening and closing and reaching full opening stage required a dark period longer than 4 h. In light-dark cycles with dark periods longer than 4 h, flower opening and closing times of the two species delayed with the delay of dawn, and the degree to which flower opening time varies with the time of dawn was greater in I. dichotoma than in I. domestica. The arrhythmia of flower opening and closing under 20L4D and LL would result from the arrhythmic output signals rather than arrhythmia of oscillators and photoreceptors. The different responses of the two species to the change of photoperiods would be caused by the transcriptional differences of genes in the output pathway of circadian clock system rather than in the input pathway or oscillators.
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Relógios Circadianos , Gênero Iris , Relógios Circadianos/genética , Ritmo Circadiano/genética , Escuridão , Flores/genética , Gênero Iris/genética , Luz , FotoperíodoRESUMO
SHORT VEGETATIVE PHASE (SVP) genes are members of the well-known MADS-box gene family that play a key role in regulating vital developmental processes in plants. Hemerocallis are perennial herbs that exhibit continuous flowering development and have been extensively used in landscaping. However, there are few reports on the regulatory mechanism of flowering in Hemerocallis. To better understand the molecular basis of floral formation of Hemerocallis, we identified and characterized the SVP-like gene HkSVP from the Hemerocallis cultivar 'Kanai Sensei'. Quantitative RT-PCR (qRT-PCR) indicated that HkSVP transcript was mainly expressed in the vegetative growth stage and had the highest expression in leaves, low expression in petals, pedicels and fruits, and no expression in pistils. The HkSVP encoded protein was localized in the nucleus of Arabidopsis protoplasts and the nucleus of onion epidermal cells. Yeast two hybrid assay revealed that HKSVP interacted with Hemerocallis AP1 and TFL1. Moreover, overexpression of HkSVP in Arabidopsis resulted in delayed flowering and abnormal phenotypes, including enriched trichomes, increased basal inflorescence branches and inhibition of inflorescence formation. These observations suggest that the HkSVP gene may play an important role in maintaining vegetative growth by participating in the construction of inflorescence structure and the development of flower organs.
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Flores/crescimento & desenvolvimento , Hemerocallis/crescimento & desenvolvimento , Proteínas de Domínio MADS/metabolismo , Proteínas de Plantas/metabolismo , Sequência de Aminoácidos , Flores/genética , Flores/metabolismo , Hemerocallis/genética , Hemerocallis/metabolismo , Inflorescência/genética , Inflorescência/crescimento & desenvolvimento , Inflorescência/metabolismo , Proteínas de Domínio MADS/genética , Filogenia , Folhas de Planta/genética , Folhas de Planta/crescimento & desenvolvimento , Folhas de Planta/metabolismo , Proteínas de Plantas/genética , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismoRESUMO
BACKGROUND: The reblooming bearded iris (Iris germanica) can bloom twice a year, in spring and autumn. The extended ornamental period makes it more popular and brings additional commercial values. However, little is known about the reblooming mechanisms, making the breeding programs time-consuming and labor-wasting. Therefore, a comparative transcriptome profiling was conducted on once-bloomers and rebloomers from the same F1 generation on six development stages, and the candidate genes associated with reblooming were identified. RESULTS: A total of 100,391 unigenes were generated, the mean length being 785 bp. In the three comparisons (the floral initiation stage of spring flowering in once-bloomers (OB-T1) vs the floral initiation stage of spring flowering in rebloomers (RB-T1); RB-T1 vs the floral initiation stage of autumn flowering in rebloomers (RB-T5); OB-T1 vs RB-T5), a total of 690, 3515 and 2941 differentially expressed genes (DEGs) were annotated against the public databases, respectively. The Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis focused on the photoperiod response, the temperature insensitivity and the growth speed, to remove the redundant DEGs and figure out the candidate key genes. As a result, the following four genes, PHYTOCHROME A (PHYA), GIGANTEA (GI), SHORT VEGETATIVE PERIOD (SVP) and AUXIN RESPONSE FACTOR (ARF), were considered to be involved in the second floral initiation of the rebloomers. CONCLUSION: This research provides valuable information for the discovery of the reblooming-related genes. The insights into the molecular mechanisms of reblooming may accelerate the breeding of bearded iris and other perennials.
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Flores , Regulação da Expressão Gênica de Plantas , Flores/genética , Perfilação da Expressão Gênica , Genes de Plantas , Iris , Anotação de Sequência Molecular , TranscriptomaRESUMO
KEY MESSAGE: The CmTFL1c gene of Chrysanthemum morifolium inhibits flowering, regulates inflorescence architecture and floral development. The timing of flowering is an important ornamental trait of chrysanthemum. The gene TERMINAL FLOWER1 (TFL1) has been shown to be involved in the regulation of meristem fate and flowering time regulation. Here, a TFL1 gene named as CmTFL1c, was cloned from Chrysanthemum morifolium and further characterized. The open reading frame of CmTFL1c comprises 522 bp, which encodes a polypeptide of 173 amino acids. Phylogenetic analysis revealed that CmTFL1c belongs to the CEN/TFL1 clade. CmTFL1c protein localizes to the nucleus as well as to plasma membrane, which suggests that CmTFL1c may be a transcription factor. The CmTFL1c gene was most highly expressed in vegetative stems, and weakly expressed in leaves and flower buds; both shoot apices and stems had sensitivity to photoperiod. Overexpression of CmTFL1c in wild Arabidopsis and tfl1-13 mutant led to late flowering and altered architecture, including increased secondary branching, and abnormal inflorescences and flowers. The CmTFL1c gene negatively regulated flowering by inhibiting the up-regulation of the AtFT, AtLFY and AtAP1. The biological function of CmTFL1c was further characterized in C. morifolium via Agrobacterium-mediated transformation, which showed that CmTFL1c not only delayed flowering and promoted axillary bud formation, but also played an important role in inflorescence formation of chrysanthemum. These results showed that the CmTFL1c affects flowering time and regulates inflorescence architecture.
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Proteínas de Arabidopsis/genética , Chrysanthemum/genética , Flores/genética , Regulação da Expressão Gênica de Plantas , Genes de Plantas/genética , Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Chrysanthemum/metabolismo , Flores/fisiologia , Perfilação da Expressão Gênica , Regulação da Expressão Gênica no Desenvolvimento , Inflorescência , Proteínas de Domínio MADS/genética , Meristema , Fases de Leitura Aberta , Fenótipo , Filogenia , Plantas Geneticamente Modificadas , Análise de Sequência de Proteína , Fatores de Transcrição/genética , Regulação para CimaRESUMO
Flowering is a very important developmental stage in the plant life cycle. LIKE HETEROCHROMATIN PROTEIN 1 (LHP1) has been shown to participate in epigenetic silencing of flowering genes. Here, for the first time, we isolated and characterized six CmLHP1 homolog genes from the important day-neutral ornamental Chrysanthemum morifolium cultivar 'Jin budiao'. These homolog genes were most likely generated by whole-genome duplication. Bioinformatic analysis showed that chrysanthemum LHP1 homologs present low similarity to other plant LHP1-like genes. However, three nuclear localization signals and two domains were highly conserved among them. The secondary structures of the CmLHP1 homologs mainly include α-helices and random coils, indicating that the proteins are mixed proteins. Phylogenetic tree analysis indicated that the six CmLHP1 genes constituted a small clade and had the closest relationship with LsLHP1 (Lactuca sativa LHP1). Quantitative RT-PCR analysis showed that the CmLHP1 homologs were expressed in different tissues during the developmental period of chrysanthemum, but they were highly expressed in the buds, especially during the key S1 stage of the inflorescence. Furthermore, the expression patterns of CmLHP1 homologs showed divergence under different photoperiods. Both CmLHP1b and CmLHP1e exhibited photoperiod sensitivity in leaves. Intriguingly, CmLHP1c was insensitive to photoperiod in both the shoot apexes and the leaves. Subcellular localization revealed that the six CmLHP1 proteins were located in the nucleus. These results reveal that CmLHP1 homolog genes could be strong candidates as important regulators of flowering time in chrysanthemum.
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Proteínas Cromossômicas não Histona/genética , Chrysanthemum/genética , Clonagem Molecular/métodos , Regulação da Expressão Gênica de Plantas , Homologia de Sequência de Aminoácidos , Motivos de Aminoácidos , Sequência de Aminoácidos , Proteínas Cromossômicas não Histona/química , Proteínas Cromossômicas não Histona/isolamento & purificação , Proteínas Cromossômicas não Histona/metabolismo , Filogenia , Proteínas de Plantas/química , Proteínas de Plantas/genética , Proteínas de Plantas/isolamento & purificação , Proteínas de Plantas/metabolismo , Estrutura Secundária de Proteína , Transporte Proteico , Frações Subcelulares/metabolismoRESUMO
We cloned and characterized the full-length coding sequence of a small heat shock (sHSP) gene, PfHSP17.2, from Primula forrestii leaves following heat stress treatment. Homology and phylogenetic analysis suggested that PfHSP17.2 is a cytosolic class II sHSP, which was further supported by the cytosolic localization of transient expression of PfHSP17.2 fused with green fluorescent protein reporter. Expression analysis showed that PfHSP17.2 was highly inducible by heat stress in almost all the vegetative and generative tissues and was expressed under salt, cold, and oxidative stress conditions as well. Moreover, the expression of PfHSP17.2 in P. forrestii was detected in certain developmental growth stages. Transgenic Arabidopsis thaliana constitutively expressing PfHSP17.2 displayed increased thermotolerance and higher resistance to salt and cold compared with wild type plants. It is suggested that PfHSP17.2 plays a key role in heat and other abiotic stresses.
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BACKGROUND: Chrysanthemum morifolium is one of the most important global cut flower and pot plants, and has been cultivated worldwide. However, limited genomic resources are available and the molecular mechanisms involved in the two morphologically distinct floret developmental cycles in chrysanthemum remain unclear. RESULTS: The transcriptomes of chrysanthemum ray florets, disc florets and leaves were sequenced using Illumina paired-end sequencing technology. In total, 16.9 G reads were assembled into 93,138 unigenes with an average length of 738 bp, of which 44,364 unigenes showed similarity to known proteins in the Swissprot or NCBI non-redundant protein databases. Additionally, 26,320, 22,304 and 13,949 unigenes were assigned to 54 gene ontology (GO) categories, 25 EuKaryotic Orthologous Groups (KOG) categories, and 280 Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways, respectively. A total of 1863 differentially expressed genes (DEGs) (1210 up-regulated and 653 down-regulated) were identified between ray florets and disc florets, including genes encoding transcription factors and protein kinases. GO and KEGG pathway enrichment analyses were performed on the DEGs to identify differences in the biological processes and pathways between ray florets and disc florets. The important regulatory genes controlling flower development and flower organ determination, as well as important functional genes in the anthocyanin biosynthetic pathway, were identified, of which two leucoanthocyanidin dioxygenase-encoding genes showed specific expression in ray florets. Lastly, reverse transcription quantitative PCR was conducted to validate the DEGs identified in our study. CONCLUSIONS: Comparative transcriptome analysis revealed significant differences in patterns of gene expression and signaling pathways between ray florets and disc florets in Chrysanthemum morifolium. This study provided the first step to understanding the molecular mechanism of the differential development of ray florets and disc florets in chrysanthemum, and also provided valuable genomic resources for candidate genes applicable for the breeding of novel varieties in chrysanthemum.
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Chrysanthemum/crescimento & desenvolvimento , Perfilação da Expressão Gênica/métodos , Proteínas de Plantas/genética , Análise de Sequência de RNA/métodos , Chrysanthemum/genética , Flores/genética , Flores/crescimento & desenvolvimento , Regulação da Expressão Gênica de Plantas , Ontologia Genética , Redes Reguladoras de Genes , Anotação de Sequência Molecular , Melhoramento Vegetal , Folhas de Planta/genética , Folhas de Planta/crescimento & desenvolvimentoRESUMO
Cardiocrinum giganteum is an endemic species of east Asia which is famous for its showy inflorescence and medicinal bulbs. Its inflorescence is a determinate raceme and the flowers bloom synchronously. Morphological observation and time-course transcriptomic analysis were combined to study the process of inflorescence and flower development of C. giganteum. The results show that the autonomic pathway, GA pathway, and the vernalization pathway are involved in the flower formation pathway of C. giganteum. A varied ABCDE flowering model was deduced from the main development process. Moreover, it was found that the flowers in different parts of the raceme in C. giganteum gradually synchronized during development, which is highly important for both evolution and ecology. The results obtained in this work improve our understanding of the process and mechanism of inflorescence and flower development and could be useful for the flowering period regulation and breeding of C. giganteum.
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Quasi-solid-state batteries (QSSBs) are gaining widespread attention as a promising solution to improve battery safety performance. However, the safety improvement and the underlying mechanisms of QSSBs remain elusive. Herein, a novel strategy combining high-safety ethylene carbonate-free liquid electrolyte and in situ polymerization technique is proposed to prepare practical QSSBs. The Ah-level QSSBs with LiNi0.83Co0.11Mn0.06O2 cathode and graphite-silicon anode demonstrate significantly improved safety features without sacrificing electrochemical performance. As evidenced by accelerating rate calorimetry tests, the QSSBs exhibit increased self-heating temperature and onset temperature (T2), and decreased temperature rise rate during thermal runaway (TR). The T2 has a maximum increase of 48.4 °C compared to the conventional liquid batteries. Moreover, the QSSBs do not undergo TR until 180 °C (even 200 °C) during the hot-box tests, presenting significant improvement compared to the liquid batteries that run into TR at 130 °C. Systematic investigations show that the in situ formed polymer skeleton effectively mitigates the exothermic reactions between lithium salts and lithiated anode, retards the oxygen release from cathode, and inhibits crosstalk reactions between cathode and anode at elevated temperatures. The findings offer an innovative solution for practical high-safety QSSBs and open up a new sight for building safer high-energy-density batteries.
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Polymethyl methacrylate (PMMA) based biomaterials have been widely utilized in clinics. However, currently, PMMA catalyzed by benzoyl peroxide (BPO) exhibits disquieting disadvantages including an exothermic polymerization reaction and a lack of bioactivity. Here, we first designed three industrial-scale synthesis methods for high-purity butoxydibutylborane (BODBB), achieving purity levels greater than 95% (maximum: 97.6%) and ensuring excellent fire safety. By utilizing BODBB as a catalyst, the highest polymerization temperature of PMMA bone cement (PMMA-BODBB) reached only 36.05 °C, ensuring that no thermal damage occurred after implantation. Compared to PMMA catalyzed by BPO and partially oxidized tributylborane (TBBO, catalyst of Super Bond C&B), PMMA-BODBB exhibited superior cell adhesion, proliferation, and osteogenesis, attributed to the reduced release of free radicals and toxic monomer, and moderate bioactive boron release. After injection into a 5 mm defect in the rat cranial bone, PMMA-BODBB demonstrated the highest level of osteointegration. This work not only presents an industrial-scale synthesis of high-purity BODBB, but also offers an innovative PMMA biomaterial system with intrinsic biocompatibility and osseointegration, paving the way for the next generation of PMMA-based biomaterials with broader applications.
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BACKGROUND: Ovarian clear cell carcinoma (OCCC), well known for its chemoresistance to platinum-based chemotherapy, exhibited a good response in clinical trials of anti-PD-1/PD-L1 inhibitors. By assessing PD-L1 expression, we sought to determine the potential therapeutic benefit of PD-1/PD-L1 inhibitors in OCCC. METHODS AND RESULTS: The retrospective study included 152 individuals with OCCC between 2019 and 2022 at Peking Union Medical College Hospital. Paired tumors of primary versus recurrent lesions (17 pairs from 15 patients) or primary versus metastatic lesions (11 pairs from 9 patients) were also included. The 22C3 pharmDx assay and whole sections were used for PD-L1 immunohistochemical staining. Pathologists with experience in premarket clinical trials evaluated PD-L1 expression based on various diagnostic criteria (TPS 1%, CPS 1, or CPS 10). The number and percentage of positive PD-L1 cases were 34 (22.4%, TPS ≥ 1%) and 59 (38.8%, CPS ≥ 1), respectively. Thirty-three (21.7%) of the cases had high PD-L1 expression (CPS ≥ 10). Half of the platinum-resistant patients (11/22) were PD-L1 positive (CPS ≥ 1). In addition, positive PD-L1 expression (CPS ≥ 1) was related to clinicopathological characteristics that represented a worse prognosis, such as advanced stages, lymph node metastasis, and distant metastasis (p = 0.032, p < 0.001 and p = 0.003, separately). PD-L1 was expressed equally or more in the recurrent lesion compared with its matched primary lesion. CONCLUSIONS: In conclusion, anti-PD-1/PD-L1 inhibitors are a promising therapeutic choice for OCCC. For evaluation of PD-L1 expression, CPS is more recommended than TPS. Evaluation of recurrent lesion was still suitable and predictive when the primary tumor tissue was not available. Distant metastatic lesions can serve as alternative samples for PD-L1 evaluation, while usage of lymphatic metastatic lesions is not recommended.
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Adenocarcinoma de Células Claras , Antígeno B7-H1 , Biomarcadores Tumorais , Neoplasias Ovarianas , Humanos , Feminino , Antígeno B7-H1/análise , Antígeno B7-H1/metabolismo , Estudos Retrospectivos , Pessoa de Meia-Idade , Neoplasias Ovarianas/patologia , Neoplasias Ovarianas/metabolismo , Neoplasias Ovarianas/tratamento farmacológico , Adulto , Idoso , Biomarcadores Tumorais/análise , Biomarcadores Tumorais/metabolismo , Adenocarcinoma de Células Claras/metabolismo , Adenocarcinoma de Células Claras/patologia , Adenocarcinoma de Células Claras/tratamento farmacológico , Imuno-Histoquímica , Resistencia a Medicamentos Antineoplásicos , Recidiva Local de Neoplasia/metabolismo , Recidiva Local de Neoplasia/patologia , Idoso de 80 Anos ou maisRESUMO
Daylily (Hemerocallis fulva) is one of the most widely used perennial flowers, but its ornamental and economic value is greatly limited due to its ephemeral flowering period. In general, the flower senescence is regulated by the developmental signals and considered as an irreversible process of programmed cell death (PCD). However, the molecular mechanism of flower PCD in daylily still remains unclear. In this study, two NAC transcription factors, namely HfNAP1 and HfNAC090, are first identified and found to be upregulated significantly in both the age-induced and the ABA-induced flower PCD processes in daylily. Then, the functions of HfNAP1 and HfNAC090 in regulating the flower PCD are investigated through transgenic phenotypes analysis. The results demonstrate that the ectopic and transient overexpression of these two genes can effectively regulate the flower PCD in tobacco and daylily. While the overexpression of HfNAP1 accelerates the flower PCD process, the overexpression of HfNAC090 significantly delays that. Furthermore, the yeast two-hybrid assay is performed to discover potential interactions related to these two genes, and the results demonstrate that HfNAP1 and HfNAC090 can interact with each other, or interact with other flower aging-related genes. Additionally, the yeast one-hybrid assay suggests that HfNAP1 and HfNAC090 can bind directly to the promoters of downstream senescence-associated genes HfSAG39 and HfSAG15. Taken overall, this study provides sufficient evidences to confirm that HfNAP1 and HfNAC090 play dominant roles in regulating the flower PCD in daylily, supporting the development of new strategies to prolong the longevity of daylily flowers.
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Reblooming bearded iris (Iris spp.) could bloom in both spring and autumn, which has extended the ornamental periods. Our previous transcriptome analysis has indicated the possible regulatory role of SHORT VEGETATIVE PHASE (SVP) in reblooming of bearded iris. Moreover, it has been revealed that the mutations of TERMINAL FLOWER 1 (TFL1) led to the continuous-flowering phenotypes in rose (Rosa spp.) and strawberry (Fragaria spp.). In order to verify the functions of these two genes on reblooming in bearded iris, IgSVP and IgTFL1 were isolated and functionally characterized. All the overexpression Arabidopsis lines of IgSVP and IgTFL1 generated the late-flowering phenotypes, indicating their functions as flowering repressors. The ectopic expression of IgSVP and IgTFL1 also generated phenotypic changes on flowers, inflorescences and branch structures. Moreover, the protein-protein interaction was found between a homologue of IgSVP and the floral meristem identity gene APETALA 1. The expression profiling showed that IgSVP was expressed significantly lower in the rebloomers in the second floral initiation stage (T5) than those of the first one (T1) in both the once-bloomers and the rebloomers, suggesting the possible regulation of IgSVP on reblooming. However, the expression level of IgTFL1 in the rebloomers was significantly higher in T5 than that in T1. The functional characterization of the two important flowering repressors IgSVP and IgTFL1 could lay solid foundation for future molecular breeding of iris, for example, knocking out the key repressors by CRISPR/Cas9 system to extend the ornamental periods of bearded iris.
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Proteínas de Arabidopsis , Arabidopsis , Proteínas de Arabidopsis/metabolismo , Flores/metabolismo , Arabidopsis/metabolismo , Inflorescência/genética , Fatores de Transcrição/metabolismo , Regulação da Expressão Gênica de PlantasRESUMO
BACKGROUND: Pulmonary invasive mucinous adenocarcinoma (IMA) is a unique type of lung adenocarcinoma with a high recurrence rate and limited treatment strategies. The tight-junction-associated protein claudin18.2 is a new therapeutic target for several solid tumors. This study aimed to detect the expression of claudin18.2 in IMA and its clinicopathological association with the disease. METHODS: The expression of claudin18.2 was immunohistochemically evaluated in an IMA cohort of 84 patients, who underwent partial pneumonectomy between January 2017 and December 2021. Positive staining for claudin18.2 was defined as ≥ 10% of tumor cells showing ≥ 1 + membrane staining or any ≥ 2 + membrane staining. RESULTS: Claudin18.2 was detected in 76.2% (64/84) of IMA patients, significantly higher than that in non-mucinous adenocarcinoma (NMA). 46.4% (39/84) of the IMA patients met the enrollment criteria of the clinical trials of monoclonal antibodies (≥ 75% of tumor cells demonstrating ≥ 2 + staining intensity). Positive staining for claudin18.2 was significantly associated with smaller tumor size (p = 0.010), less pleural invasion (p = 0.019), and earlier pN stage (p < 0.001). Expression of claudin18.2 was not related to prognosis in multivariate analysis. CONCLUSIONS: To summarize, in this study we found that claudin18.2 was remarkably highly expressed in IMA and the overexpression was associated with low invasive capacity. Thus, this protein appears to be a promising therapeutic target and deserves further investigation in IMA patients.
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AIM: As the second most prevalent subtype of epithelial ovarian cancers, ovarian clear cell carcinoma (OCCC) is known for its chemoresistance to conventional platinum-based therapy. In this work, we examined the tryptophan (Trp) metabolism enzymes' differential expression in patients with OCCC to assess the potential for personalised treatment. METHODS: A total of 127 OCCC tissues were used to construct tissue microarrays, and immunohistochemistry (IHC) staining of the Trp enzymes IDO1, IDO2, TDO2 and IL4I1 was performed. The correlations between Trp enzyme expression and clinical characteristics were analysed. RESULTS: Positive IDO1, IDO2, TDO2 and IL4I1 staining was identified in 26.8%, 94.5%, 75.6% and 82.7% of OCCC respectively. IDO1-positive samples were more common in the chemoresistant group than in the platinum-sensitive group (46.7% vs. 19.8%). Moreover, positive expression of IDO1, TDO2 and IL4I1 was related to advanced stage, metastasis, bilateral tumours, endometriosis and tumour rupture (p < 0.05) respectively. Univariate analysis revealed a significant association between bilateral tumours, lymph node metastasis, advanced stage, distant metastasis and aberrant cytology with a poor prognosis for OCCC, while the absence of residual tumour was correlated with a favourable outcome (p < 0.05). However, only bilateral tumours and lymph node metastases were related to a poor prognosis after multivariate analysis. CONCLUSION: This is the first study to investigate the expression of the Trp enzymes IDO1, IDO2, TDO2 and IL4I1 in OCCC tissues. IDO2, TDO2 and IL4I1 were detected in the majority of OCCC. Clinical traits were correlated with IDO1, IDO2, TDO2 and IL4I1 expression. IDO1 may be used as a therapeutic target given the large percentage of chemoresistant cases with IDO1 expression. These results will aid the development of personalised therapies for OCCC.
Assuntos
Neoplasias Ovarianas , Triptofano , Feminino , Humanos , Triptofano/metabolismo , Triptofano Oxigenase , Carcinoma Epitelial do Ovário , Neoplasias Ovarianas/patologia , L-Aminoácido OxidaseRESUMO
BACKGROUND: Metabolites of tryptophan (Trp) metabolism in the tumor microenvironment play crucial immunosuppressive roles in various cancers. However, the role of Trp metabolism in diffuse large B-cell lymphoma (DLBCL) or natural killer/T-cell lymphoma (NK/TCL) remains unelucidated. METHODS: We investigated the potential role of Trp metabolism in a cohort of 43 patients with DLBCL and 23 with NK/TCL. We constructed tissue microarrays and performed in situ staining of Trp-catabolizing enzymes and PD-L1 using immunohistochemistry (IHC). RESULTS: We observed 14.0% positive staining of IDO1 in DCBCL and 60.9% in NK/TCL; 55.8% of IDO2 in DCBCL and 95.7% in NK/TCL; 79.1% of TDO2 in DCBCL and 43.5% in NK/TCL; 29.7% of IL4I1 in DCBCL and 39.1% in NK/TCL. However, IDO1, IDO2, TDO2, and IL4I1 positivity did not significantly differ between PD-L1+ and PD-L1- biopsy tissue samples of NK/TCL; nonetheless, a positive correlation of IDO1 (r = 0.87, p < 0.001), IDO2 (r = 0.70, p < 0.001), TDO2 (r = 0.63, p < 0.001), and IL4I1 (r = 0.53, p < 0.05) with PD-L1 expression was observed in the TCGA-DLBCL dataset. Finally, immunohistochemical (IHC) analysis revealed the lack of superior prognostic effect with higher expression of Trp enzymes in DLBCL and NK/TCL. Furthermore, IDO1, IDO2, TDO2, and IL4I1 expression, as well as survival rates, did not significantly differ across all groups in the TCGA-DLBCL cohort. CONCLUSION: Collectively, our findings provide novel insights into the enzymes involved in Trp metabolism in DLBCL and NK/TCL and their association with PD-L1 expression, which offers potential strategies to combine Trp-metabolism enzyme inhibitors with anti-PD-L1 or other immunotherapeutic strategies in clinical DLBCL or NK/TCL treatment.
Assuntos
Linfoma Difuso de Grandes Células B , Linfoma de Células T , Humanos , Triptofano/uso terapêutico , Prognóstico , Linfoma Difuso de Grandes Células B/tratamento farmacológico , Imuno-Histoquímica , Antígeno B7-H1/metabolismo , Microambiente Tumoral , L-Aminoácido Oxidase/uso terapêuticoRESUMO
The two most prevalent subtypes of epithelial ovarian carcinoma (EOC) are ovarian clear cell carcinoma (OCCC) and high-grade serous ovarian carcinoma (HGSC). Patients with OCCC have a poor prognosis than those with HGSC due to chemoresistance, implying the need for novel treatment target. In this study, we applied single-cell RNA sequencing (scRNA-seq) together with bulk RNA-seq data from the GEO (Gene Expression Omnibus) database (the GSE189553 dataset) to characterize and compare tumor heterogeneity and cell-level evolution between OCCC and HGSC samples. To begin, we found that the smaller proportion of an epithelial OCCC cell subset in the G2/M phase might explain OCCC chemoresistance. Second, we identified a possible pathogenic OCCC epithelial cell subcluster that overexpresses LEFTY1. Third, novel biomarkers separating OCCC from HGSC were discovered and subsequently validated on a wide scale using immunohistochemistry. Amine oxidase copper containing 1 (AOC1) was preferentially expressed in OCCC over HGSC, while S100 calcium-binding protein A2 (S100A2) was detected less frequently in OCCC than in HGSC. In addition, we discovered that metabolic pathways were enriched in the epithelial compartment of the OCCC samples. In vitro experiments verified that inhibition of oxidative phosphorylation or glycolysis pathways exerted direct antitumor effects on both OCCC and HGSC cells, while targeting glutamine metabolism or ferroptosis greatly attenuated chemosensitivity only in OCCC cells. Finally, to determine whether there were any variations in immune cell subsets between OCCC and HGSC, data from scRNA-seq and mass cytometry were pooled for analysis. In summary, our work provides the first holistic insights into the cellular and molecular distinctions between OCCC and HGSC and is a valuable source for discovering new targets to leverage in clinical treatments to improve the poor prognosis of patients with OCCC.