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Rice blast, caused by Pyricularia oryzae, is one of the most destructive rice diseases worldwide. Using resistant rice varieties is the most cost-effective way to control rice blast. Consequently, it is critical to monitor the distribution frequency of avirulence (Avr) genes in rice planting fields to facilitate the breeding of resistant rice varieties. In this study, we established a rapid recombinase polymerase amplification-lateral flow dipstick (RPA-LFD) detection system for the identification of AvrPik, Avr-Piz-t, and Avr-Pi9. The optimized reaction temperature and duration were 37°C and 20 min, indicating that the reaction system could be initiated by body temperature without relying on any precision instruments. Specificity analysis showed that the primer and probe combinations targeting the three Avr genes exhibited a remarkable specificity at genus-level detection. Under the optimized condition, the lower detected thresholds of AvrPik, Avr-Piz-t, and Avr-Pi9 were 10 fg/µl, 100 fg/µl, and 10 pg/µl, respectively. Notably, the detection sensitivity of the three Avr genes was much higher than that of PCR. In addition, we also successfully detected the presence of AvrPik, Avr-Piz-t, and Avr-Pi9 in the leaf and panicle blast lesions with the RPA-LFD detection system. In particular, the genomic DNA was extracted using the simpler PEG-NaOH rapid extraction method. In summary, we developed an RPA detection system for AvrPik, Avr-Pi9, and Avr-Piz-t, combined with the PEG-NaOH rapid DNA extraction method. The innovative approach achieved rapid, real-time, and accurate detection of the three Avr genes in the field, which is helpful to understand the distribution frequency of the three Avr genes in the field and provide theoretical reference for the scientific layout of resistant rice varieties.
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Ascomicetos , Oryza , Enfermedades de las Plantas , Enfermedades de las Plantas/microbiología , Oryza/microbiología , Ascomicetos/genética , Ascomicetos/patogenicidad , Técnicas de Amplificación de Ácido Nucleico/métodos , Virulencia/genética , Genes Fúngicos/genéticaRESUMEN
Single-cell sequencing (scRNA-seq) has revolutionized our ability to explore heterogeneity and genetic variations at the single-cell level, opening up new avenues for understanding disease mechanisms and cell-cell interactions. Single-nucleus RNA-sequencing (snRNA-seq) is emerging as a promising solution to scRNA-seq due to its reduced ionized transcription bias and compatibility with richer samples. This approach will provide an exciting opportunity for in-depth exploration of billions of formalin-fixed paraffin-embedded (FFPE) tissues. Recent advancements in single-cell/nucleus gene expression workflows tailored for FFPE tissues have demonstrated their feasibility and provided crucial guidance for future studies utilizing FFPE specimens. In this review, we provide a broad overview of the nuclear preparation strategies, the latest technologies of snRNA-seq applicable to FFPE samples. Finally, the limitations and potential technical developments of snRNA-seq in FFPE samples are summarized. The development of snRNA-seq technologies for FFPE samples will lay a foundation for transcriptomic studies of valuable samples in clinical medicine and human sample banks.
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High prolactin (PRL) concentration has been shown to induce the apoptosis of ovine ovarian granulosa cells (GCs), but the underlying mechanisms are unclear. This study aimed to investigate the mechanism of apoptosis induced by high PRL concentration in GCs. Trial 1: The optimal concentration of glutathion was determined according to the detected cell proliferation. The results showed that the optimal glutathione concentration was 5 µmol/mL. Trial 2: 500 ng/mL PRL was chosen as the high PRL concentration. The GCs were treated with 0 ng/mL PRL (C group), 500 ng/mL PRL (P group) or 500 ng/mL PRL, and 5 µmol/mL glutathione (P-GSH group). The results indicated that the mitochondrial respiratory chain complex (MRCC) I-V, ATP production, total antioxidant capacity (T-AOC), superoxide dismutase (SOD), and thioredoxin peroxidase (TPx) in the C group were higher than those in the P group (p < 0.05), while they were lower than those in the P-GSH group (p < 0.05). Compared to the C group, the P group exhibited elevated levels of reactive oxygen species (ROS) and apoptosis (p < 0.05) and increased expression of ATG7 and ATG5 (p < 0.05). However, MRCC I-V, ATP, SOD, A-TOC, TPx, ROS, and apoptosis were decreased after the addition of glutathione (p < 0.05). The knockdown of either L-PRLR or S-PRLR in P group GCs resulted in a significant reduction (p < 0.05) in MRCC I-V, ATP, T-AOC, SOD and TPx, while the overexpression of either receptor showed an opposite trend (p < 0.05). Our findings suggest that high PRL concentrations induce apoptotic cell death in ovine ovarian GCs by downregulating L-PRLR and S-PRLR, activating oxidative stress and autophagic pathways.
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Prolactina , Receptores de Prolactina , Femenino , Animales , Ovinos , Prolactina/farmacología , Prolactina/metabolismo , Receptores de Prolactina/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Estrés Oxidativo , Apoptosis , Antioxidantes/metabolismo , Células de la Granulosa/metabolismo , Glutatión/metabolismo , Superóxido Dismutasa/metabolismo , Adenosina Trifosfato/metabolismoRESUMEN
INTRODUCTION: In recent years, China's economy has grown rapidly, and the health condition of Chinese residents has significantly improved. However, this rapid economic and social development has also brought a series of environmental problems, such as serious haze pollution, of which the main contents are PM2.5 particles. The objective of this study is to quantitatively estimate the PM2.5 -related health costs in China. METHODS: Based on city-level data from 140 major Chinese cities as well as the Beijing-Tianjin-Hebei, Yangtze River Delta, and Pearl River Delta city clusters in 2010, the value of a statistical life method based on willingness to pay was employed. Moreover, global and local Moran's I values were calculated to examine the spatial distribution of the health cost of haze pollution in China. RESULTS: In areas with heavy haze pollution or a high level of economic development, residents' health costs will also be higher. In addition, there is a spatial aggregation phenomenon in the spatial distribution of health costs in China, which is mainly in the form of "high-high" aggregation, with high-value cities converging with other high-value cities. CONCLUSIONS: The health cost of haze pollution in China is very considerable, and there are regional differences.
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Contaminantes Atmosféricos , Contaminación del Aire , Contaminación del Aire/efectos adversos , China , Ciudades , Costos de la Atención en Salud , Material Particulado/análisisRESUMEN
Long non-coding RNAs (lncRNAs) have been shown to play important roles in livestock fecundity, and many lncRNAs that affect follicular development and reproductive diseases have been identified in the ovary. However, only a few of them have been functionally annotated and mechanistically validated. In this study, we identified a new lncRNA (lncGSAR) and investigated its effects on the proliferation and steroidogenesis of ovine granulosa cells (GCs). High concentrations of glucose (add 33.6 mM glucose) caused high expression of lncGSAR in GCs by regulating its stability, and lncGSAR overexpression promoted GCs proliferation, estrogen secretion, and inhibited progesterone secretion, whereas interference with lncGASR had the opposite effect. Next, we found that the RNA molecules of lncGSAR act on MiR-125b as competitive endogenous RNA (ceRNA), and SREBP-cleavage-activating protein (SCAP) was verified as a target of MiR-125b. LncGASR overexpression increased the expression of SCAP, SREBP, and steroid hormone-related proteins, which can be attenuated by MiR-125b. Our results demonstrated that lncGSAR can act as a ceRNA to activate SCAP/SREBP signaling by sponging MiR-125b to regulate steroid hormone secretion in GCs. These findings provide new insights into the mechanisms of nutrient-regulated follicle development in ewes.
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MicroARNs , ARN Largo no Codificante , Ovinos/genética , Animales , Femenino , ARN Largo no Codificante/genética , ARN Largo no Codificante/metabolismo , Proteína 1 de Unión a los Elementos Reguladores de Esteroles/metabolismo , MicroARNs/genética , MicroARNs/metabolismo , Progesterona/metabolismo , Células de la Granulosa/metabolismo , Estrógenos/metabolismo , Glucosa/metabolismo , Proliferación Celular/genéticaRESUMEN
Short-term dietary supplementation of ewes during the luteal phase can increase fertility, most probably by stimulating glucose uptake by the follicles. However, the molecular mechanism of glucose regulation of follicular development has not yet been clarified, especially the further study of long non-coding RNA (lncRNA) in determining fertility during follicular development. We generated granulosa cell (GC) models of different doses of glucose (0, 2.1, 4.2, 8.4, 16.8 and 33.6 mM), and observed that the highest cell viability was recorded in the 8.4 mM group and the highest apoptosis rates were recorded in the 33.6 mM group. Therefore, a control group (n = 3, 0 mM glucose), a low glucose group (n = 3, add 8.4 mM glucose), and a high glucose group (n = 3, add 33.6 mM glucose) of GCs were created for next whole genomic RNA sequencing. In total, 18,172 novel lncRNAs and 510 annotated lncRNAs were identified in the GCs samples. Gene Ontology indicated that differentially expressed lncRNAs associated with cell apoptosis were highly enriched. Kyoto Encyclopedia of Genes and Genomes enrichment analysis of lncRNA target genes found that the apoptosis pathway and the p53 signaling pathway were both enriched. Furthermore, we focused on the function of a lncGDAR and verified that lncGDAR could influence cell apoptosis in GC development through affecting the mRNA and protein expression of apoptosis-related markers. These results provide the basis for further study of the lncRNA regulation mechanism in nutrition on female fertility.
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ARN Largo no Codificante , Animales , Apoptosis/genética , Femenino , Perfilación de la Expresión Génica , Glucosa/metabolismo , Células de la Granulosa/metabolismo , ARN Largo no Codificante/genética , ARN Largo no Codificante/metabolismo , ARN Mensajero/genética , Ovinos/genéticaRESUMEN
Parkinson's disease (PD) is a neurodegenerative disease with an impairment of movement execution that is related to age and genetic and environmental factors. 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) is a neurotoxin widely used to induce PD models, but the effect of MPTP on the cells and genes of PD has not been fully elucidated. By single-nucleus RNA sequencing, we uncovered the PD-specific cells and revealed the changes in their cellular states, including astrocytosis and endothelial cells' absence, as well as a cluster of medium spiny neuron cells unique to PD. Furthermore, trajectory analysis of astrocyte and endothelial cell populations predicted candidate target gene sets that might be associated with PD. Notably, the detailed regulatory roles of astrocyte-specific transcription factors Dbx2 and Sox13 in PD were revealed in our work. Finally, we characterized the cell-cell communications of PD-specific cells and found that the overall communication strength was enhanced in PD compared with a matched control, especially the signaling pathways of NRXN and NEGR. Our work provides an overview of the changes in cellular states of the MPTP-induced mouse brain.
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Intoxicación por MPTP , Enfermedades Neurodegenerativas , Enfermedad de Parkinson , 1-Metil-4-fenil-1,2,3,6-Tetrahidropiridina/efectos adversos , Animales , Modelos Animales de Enfermedad , Células Endoteliales/metabolismo , Intoxicación por MPTP/genética , Intoxicación por MPTP/metabolismo , Ratones , Ratones Endogámicos C57BL , Neurotoxinas/efectos adversos , Enfermedad de Parkinson/genética , Enfermedad de Parkinson/metabolismo , Análisis de Secuencia de ARN , Factores de Transcripción/genéticaRESUMEN
Environmental emergencies are associated with great harm, sudden onset and high publicity and cause serious damage to a broad range of ecological environmental, human health and social properties in a short period of time. Environmental decentralization can help local governments strengthen their independence and make better use of the advantages of environmental information to curb the occurrence of environmental emergencies, but regional corruption significantly weakens its effectiveness. Therefore, based on panel data of China's 30 provincial administrative regions between 2005 and 2016, this paper applies the panel threshold model to explore the relationship between environmental emergencies, environmental decentralization, and regional corruption. The results indicate that, first, environmental decentralization, environmental administrative decentralization, environmental monitoring decentralization, and environmental supervision decentralization all have a negative influence on environmental emergencies; second, as the degree of corruption in a region increases, the effect of environmental decentralization on restraining environmental emergencies decreases; and finally, there is heterogeneity in the relationship between environmental emergencies and environmental decentralization in various regions. Environmental decentralization in the eastern and western regions negatively affects environmental emergencies, while there is a positive relationship between the two in the central region.
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Urgencias Médicas , Gobierno Local , China , Humanos , PolíticaRESUMEN
In the absence of pathogen attack, lesion mimic mutants (LMMs) in plants undergo spontaneous cell death and develop necrosis or apoptosis-like lesions on the leaves or sheath, resembling symptoms of hypersensitive response. In-depth research has been conducted on LMMs, especially regarding the molecular mechanisms underlying programmed cell death and disease resistance. In this study, the spotted leaf 36 (spl36) mutant was identified as a typical LMM, showing lesions on both the leaf blade and leaf sheath. The formation of lesions was found to be caused by cell death accompanied by accumulation of hydrogen peroxide and degradation of chloroplasts. Compared with wild-type, the main agronomic traits such as plant height, effective panicle number, panicle length, grain per panicle, seed setting rate, and 1000-grain weight of spl36 were significantly reduced. The defence and pathogenesis-related genes PR1a, PR1b, PR10, and NPR1, were transcriptionally activated in mutant spl36 without pathogen attack. Genetic analysis showed that the mutant phenotype was controlled by the gene SPL36, which was mapped to an interval of 260 kb at the end of the long arm on chromosome 11. Pathogen inoculation analysis showed that spl36 has enhanced resistance to sheath blight, rice blast, and bacterial blight.
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Zooxanthellae and coral can form an intracellular symbiotic system. Yet, little is known about the molecular mechanism underlying this symbiosis. In this study, we characterized the symbiosis based on analyses of gene expression at the single-cell level. Among 9110 single coral cells, we identified 4871 symbiotic cells based on the detection of both coral and zooxanthellae gene transcripts within a single cell. Using the bioinformatics tool Seurat, symbiotic cells were further clustered into five groups, 52 genes exhibited differential expression between groups. We proposed an index called the "symbiosis index", to indicate the degree of gene expression of both species in a single symbiotic cell. Interestingly, the index differed distinctly among the five groups. The symbiosis index was highly correlated with the expression of the coral gene gfas1.m1.6761 (ANKRD40), which encodes ankyrin repeat domain-containing protein 40 and is involved in DNA replication (r = 0.76). Two metabolism-related genes, DAGLA and betaGlu, were highly expressed in cells with a high symbiosis index. Four zooxanthellae genes, PRPF19, ATRN, aAA-ATPases and AK812-SmicGene44833, exhibited substantial changes in expression levels when zooxanthellae lived within coral. A trajectory analysis suggested that cells with a higher symbiosis index may be derived from those with a lower index during coral colony development. Taken together, our results provide evidence for zooxanthellae residing within coral, forming a symbiotic system. The symbiosis index is an effective indicator of different cell groups, with lineage relationships among groups. Additionally, we identified specific genes that exhibit expression changes in the symbiotic system.
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Antozoos/genética , Dinoflagelados/genética , Simbiosis/genética , Animales , Antozoos/metabolismo , Análisis por Conglomerados , Dinoflagelados/metabolismo , RNA-Seq , Análisis de la Célula IndividualRESUMEN
AMP-activated protein kinase (AMPK) is a metabolic energy sensor that plays a critical role in cancer cell survival and growth. While the physical microenvironment is believed to influence tumor growth and progression, its role in AMPK regulation remains largely unknown. In the present study, we evaluated AMPK response to mechanical forces and its interaction with other mechano-responsive signaling proteins, FAK and Src. Using genetically encoded biosensors that can detect AMPK activities at different subcellular locations (cytosol, plasma membrane, nucleus, mitochondria, and Golgi apparatus), we observed that AMPK responds to shear stress in a subcellular location-dependent manner in breast cancer cells (MDA-MB-231). While normal epithelial cells (MCF-10A) also similarly responded to shear stress, they are less sensitive to shear stress compared to MDA-MB-231 cells. Inhibition of FAK and Src significantly decreased the basal activity level of AMPK at all five subcellular locations in MDA-MB-231 cells and selectively blocked shear stress-induced AMPK activation. Moreover, testing with cytoskeletal drugs revealed that myosin II might be the critical mediator of shear stress-induced AMPK activation in MDA-MB-231 cells. These findings suggest that breast cancer cells and normal epithelial cells may have different mechanosensitivity in AMPK signaling and that FAK and Src as well as the myosin II-dependent signaling pathway are involved in subcellular AMPK mechanotransduction in breast cancer cells.
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Proteínas Quinasas Activadas por AMP/metabolismo , Quinasa 1 de Adhesión Focal/metabolismo , Hidrodinámica , Espacio Intracelular/metabolismo , Mecanotransducción Celular , Familia-src Quinasas/metabolismo , Línea Celular Tumoral , Citoesqueleto/metabolismo , Activación Enzimática , Humanos , Resistencia al Corte , Estrés MecánicoRESUMEN
Health problems caused by environmental pollution may affect the process of urbanization in China. Therefore, this study, against the backdrop of promoting new-type urbanization, evaluates the level of China's urbanization comprehensively using the fully arranged polygon graphical index method. It uses a dynamic threshold panel model to study the potential non-linear relationship between environmental pollution (wastewater, sulfur dioxide, and solid wastes) and urbanization under different health costs of residents. Our findings show that environmental pollution has inhibited the improvement of comprehensive urbanization, population urbanization, economic urbanization, and living conditions urbanization, but promoted living environment urbanization, in China. It is worth noting that with the rise in residents' health costs, the inhibiting effect of environmental pollution on comprehensive urbanization, population urbanization, economic urbanization, and living conditions urbanization in China has gradually increased, but on living environment urbanization, it has decreased.
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Contaminación Ambiental , Costos de la Atención en Salud , Urbanización , China , Humanos , Dióxido de AzufreRESUMEN
PURPOSE: To review the clinical presentation, treatment, and prognosis of patients with malignant mixed tumor (carcinoma ex pleomorphic adenoma) of the lacrimal gland. METHODS: Clinical records and radiographic images were reviewed for patients with malignant mixed tumor of the lacrimal gland treated at the center during 2008-2019. RESULTS: The study included 9 patients (6 men, 3 women) aged 17-66 years (median age, 56 years). Six had primary malignant mixed tumor with no history of orbital lesions, and 3 had previously been diagnosed with pleomorphic adenoma. Tumor, Node, Metastasis classification per the eighth edition of the American Joint Committee on Cancer (AJCC) Cancer Staging Manual were T1aN0M0 in 2 patients, T2aN0M0 in 3 patients, T4bN0M0 in 2 patients, and T4cN0M0 in 2 patients. Two patients underwent orbital exenteration, 6 patients underwent eye-sparing surgery, and 1 patient had an unresectable tumor because of cavernous sinus extension. All patients received radiotherapy (intensity-modulated radiotherapy in 3 and proton therapy in 6). All patients received chemotherapy, 8 concurrently with radiotherapy and 1 after radiotherapy. The median follow-up time was 70 months. At last contact, 6 patients were alive without evidence of disease; 2 had died of disease, 1 of distant metastasis, and the other of cavernous sinus invasion. CONCLUSIONS: The findings suggest that de novo malignant mixed tumor of the lacrimal gland is more common than disease that results from transformation after incomplete resection of lacrimal gland pleomorphic adenoma. Most cases can be treated with eye-sparing surgery and radiation unless skull base extension is present.
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Adenoma Pleomórfico , Carcinoma , Neoplasias del Ojo , Enfermedades del Aparato Lagrimal , Aparato Lagrimal , Tumor Mixto Maligno , Adolescente , Adulto , Anciano , Neoplasias del Ojo/diagnóstico , Neoplasias del Ojo/terapia , Femenino , Humanos , Aparato Lagrimal/diagnóstico por imagen , Enfermedades del Aparato Lagrimal/diagnóstico , Enfermedades del Aparato Lagrimal/terapia , Masculino , Persona de Mediana Edad , Adulto JovenRESUMEN
The biophysical microenvironment of the tumor site has significant impact on breast cancer progression and metastasis. The importance of altered mechanotransduction in cancerous tissue has been documented, yet its role in the regulation of cellular metabolism and the potential link between cellular energy and cell migration remain poorly understood. In this study, we investigated the role of mechanotransduction in AMP-activated protein kinase (AMPK) activation in breast cancer cells in response to interstitial fluid flow (IFF). Additionally, we explored the involvement of AMPK in breast cancer cell migration. IFF was applied to the 3D cell-matrix construct. The subcellular signaling activity of Src, FAK, and AMPK was visualized in real-time using fluorescent resonance energy transfer (FRET). We observed that breast cancer cells (MDA-MB-231) are more sensitive to IFF than normal epithelial cells (MCF-10A). AMPK was activated at the mitochondria of MDA-MB-231â¯cells by IFF, but not in other subcellular compartments (i.e., cytosol, plasma membrane, and nucleus). The inhibition of FAK or Src abolished flow-induced AMPK activation in the mitochondria of MDA-MB-231â¯cells. We also observed that global AMPK activation reduced MDA-MB-231â¯cell migration. Interestingly, specific AMPK inhibition in the mitochondria reduced cell migration and blocked flow-induced cell migration. Our results suggest the linkage of FAK/Src and mitochondria-specific AMPK in mechanotransduction and the differential role of AMPK in breast cancer cell migration depending on its subcellular compartment-specific activation.
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Proteínas Quinasas Activadas por AMP/genética , Células Epiteliales/enzimología , Quinasa 1 de Adhesión Focal/genética , Regulación Neoplásica de la Expresión Génica , Mecanotransducción Celular/genética , Mitocondrias/enzimología , Familia-src Quinasas/genética , Proteínas Quinasas Activadas por AMP/antagonistas & inhibidores , Proteínas Quinasas Activadas por AMP/metabolismo , Compuestos de Bifenilo , Línea Celular , Línea Celular Tumoral , Movimiento Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Células Epiteliales/efectos de los fármacos , Células Epiteliales/patología , Transferencia Resonante de Energía de Fluorescencia , Quinasa 1 de Adhesión Focal/antagonistas & inhibidores , Quinasa 1 de Adhesión Focal/metabolismo , Humanos , Glándulas Mamarias Humanas , Mitocondrias/efectos de los fármacos , Mitocondrias/patología , Especificidad de Órganos , Pirimidinas/farmacología , Pironas/farmacología , Quinolonas/farmacología , Reología , Estrés Mecánico , Sulfonas/farmacología , Tiofenos/farmacología , Microambiente Tumoral/genética , Familia-src Quinasas/antagonistas & inhibidores , Familia-src Quinasas/metabolismoRESUMEN
PURPOSE: To evaluate the frequency and nature of changes in T category when eyelid carcinomas are staged using the criteria in the 8th edition instead of the 7th edition of the American Joint Committee on Cancer staging manual. METHODS: Following Institutional Review Board approval, a retrospective review was conducted for all consecutive patients with the diagnosis of eyelid carcinoma treated by the senior author from January 2012 through December 2016. After a review of the clinical and pathologic data, each patient's disease was staged using both the 7th-edition and 8th-edition American Joint Committee on Cancer criteria for eyelid carcinomas. Changes in T categories between the 2 staging systems were examined. RESULTS: The review initially identified 167 patients with the diagnosis of eyelid carcinoma. Four patients were excluded because of incomplete or unclear data. The remaining 163 patients included 78 men and 85 women aged 21 to 97 years (median, 68 years). Eighty-two patients had basal cell carcinoma; 35, squamous cell carcinoma; 32, sebaceous carcinoma; 6, mucinous eccrine carcinoma; 3, Merkel cell carcinoma; 3, adenocarcinomas; and 2, adnexal carcinoma. The most common T category according to the 7th-edition criteria was T2a; the most common T category according to the 8th-edition criteria was T1b. Of the 163 patients, 64 (39%) had a lower T category with the 8th-edition than with the 7th-edition criteria, 59 (36%) had a higher T category, and 40 (25%) had the same T category. CONCLUSIONS: Application of the 8th-edition American Joint Committee on Cancer criteria for eyelid carcinoma changed the T category in 75% of patients. In general, the new 8th-edition American Joint Committee on Cancer tumor, node, metastasis (TNM) designations allow for a more objective and consistent designation of the T category.
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Neoplasias de los Párpados/patología , Guías como Asunto , Oncología Médica , Estadificación de Neoplasias/métodos , Sociedades Médicas , Adulto , Anciano , Anciano de 80 o más Años , Femenino , Humanos , Masculino , Persona de Mediana Edad , Reproducibilidad de los Resultados , Estudios Retrospectivos , Estados Unidos , Adulto JovenRESUMEN
PURPOSE: A risk assessment score for metastasis based on age, tumor size, and mitotic figures has been suggested for nonorbital solitary fibrous tumor (SFT)/hemangiopericytoma. The authors herein examine the clinicopathological features of recurrent and metastatic orbital SFT and evaluate the existing risk assessment score for orbital SFT. METHODS: The American Society of Ophthalmic Plastic and Reconstructive Surgery Oncology Database was queried for patients with recurrent or malignant orbital hemangiopericytoma/SFT. The medical records were reviewed for clinical and pathologic findings, treatments, and outcomes. RESULTS: Eight patients from 3 institutions were identified with recurrent orbital hemangiopericytoma/SFT. Median age at diagnosis was 59 years, and 4 patients were women. The mean size of tumor was 2.1 ± 1.1 cm. All patients were initially treated with surgery and experienced local recurrence after a median of 4 (range 0.5-10) years. Five patients were treated with orbital radiation. Two patients also developed distant metastases and eventually died of their disease. Median Ki-67 was 5% (range 1-65%) and 5 mitotic figures/10 high-power fields (range 2-30). The previously described risk stratification model for nonorbital SFT did not correlate with the propensity to develop metastases in this cohort; however, both patients with distant metastasis had > 4 mitotic figures /10 high-power fields. CONCLUSIONS: In this cohort of recurrent orbital hemangiopericytoma/SFT, median time to recurrence was 4 years underscoring the importance of careful continued follow-up. The current risk stratification models have limited use for orbital lesions, mostly due to the fact that orbital SFTs are smaller than even the smallest size criteria in this risk assessment model.
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Hemangiopericitoma/patología , Recurrencia Local de Neoplasia/patología , Neoplasias Orbitales/patología , Tumores Fibrosos Solitarios/patología , Adulto , Anciano , Diagnóstico Diferencial , Femenino , Estudios de Seguimiento , Hemangiopericitoma/cirugía , Humanos , Incidencia , Imagen por Resonancia Magnética , Masculino , Persona de Mediana Edad , Recurrencia Local de Neoplasia/epidemiología , Procedimientos Quirúrgicos Oftalmológicos , Neoplasias Orbitales/cirugía , Estudios Retrospectivos , Tumores Fibrosos Solitarios/cirugía , Factores de Tiempo , Estados Unidos/epidemiologíaRESUMEN
OBJECTIVE: To examine the microvascular pathological characteristics and changes in related injury factors in a rat model of acute blood stasis. METHODS: A total of 75 Sprague-Dawley rats were divided randomly and equally into a control group and four experimental groups assessed at different times after the induction of stasis (0, 1, 3 or 6 h after stasis) (n = 15). The acute blood stasis model was established through rat tail-vein injection of high-molecular-weight dextran. After Electrocardiograph (ECG) detection at predetermined times (0, 1, 3 and 6 h after induction of stasis), the rats were sacrificed and blood and cardiac samples were harvested for analysis. Hematoxylin-eosin (HE) staining and transmission electron microscopy were used for histopathological detection; an enzyme linked immunosorbent assay (ELISA) was used to detect thromboxane B2 (TXB2) and 6-keto-prostaglandin F1α (6-Keto-PGF1α) concentrations; a real-time polymerase chain reaction (PCR) reaction system was used to detect intercellular adhesion molecule 1 (ICAM-1) and vascular cell adhesion molecule1 (VCAM-1) mRNA expression; western blotting was used to detect vascular endothelial cadherin (VE-cadherin) protein expression. RESULTS: The ST segment in the ECG showed gradual elevation after induction of stasis and continued elevation at a high level at 3 and 6 h. The HE staining showed changes in myocardial cell necrosis and tissue dissociation after the induction of stasis, along with inflammatory infiltration. Results of transmission electron microscopy showed immediate changes in blood stasis and lumen occlusion in the microvasculature, along with endothelial cell swelling. After the induction of stasis, TXB2 concentrations gradually increased while 6-Keto-PGF(1α) concentrations were immediately significantly reduced. The TXB(2)/6-Keto-PGF(1α) ratio was maintained at a high level. ICAM-1 mRNA expression showed an unstable elevation while VCAM-1 mRNA expression was significantly reduced after the induction of stasis. Compared with the control group, VE-cadherin protein expression increased at 0 and 3 h after the induction of stasis, while no change occurred at 1 and 6 h. CONCLUSION: The pathological manifestations of acute blood stasis are microvascular blood retention, lumen stenosis and even occlusion. The condition is also called "blood coagulation and weep" in Traditional Chinese Medicine. The blood stasis model resulted in the injury and necrosis of endothelial cells and cardiomyocytes, along with the presence of an imbalance of vasomotor factor levels, platelet activation, and increases in the expression of adhesion molecules and endothelial barrier dysfunction, which corresponds to "blood failed to nourish" in Traditional Chinese Medicine.
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Infarto del Miocardio/patología , 6-Cetoprostaglandina F1 alfa/sangre , Animales , Moléculas de Adhesión Celular/sangre , Modelos Animales de Enfermedad , Electrocardiografía , Corazón/fisiopatología , Humanos , Molécula 1 de Adhesión Intercelular/metabolismo , Masculino , Microvasos/metabolismo , Microvasos/patología , Microvasos/fisiopatología , Infarto del Miocardio/genética , Infarto del Miocardio/metabolismo , Infarto del Miocardio/fisiopatología , Miocardio/metabolismo , Miocardio/patología , Ratas , Ratas Sprague-Dawley , Tromboxano B2/sangre , Molécula 1 de Adhesión Celular Vascular/genética , Molécula 1 de Adhesión Celular Vascular/metabolismoRESUMEN
The combination of single-cell RNA sequencing and microdissection techniques that preserves positional information has become a major tool for spatial transcriptome analyses. However, high costs and time requirements, especially for experiments at the single cell scale, make it challenging for this approach to meet the demand for increased throughput. Therefore, we proposed combinational DNA barcode (CDB)-seq as a medium-throughput, multiplexed approach combining Smart-3SEQ and CDB magnetic microbeads for transcriptome analyses of microdissected tissue samples. We conducted a comprehensive comparison of conditions for CDB microbead preparation and related factors and then applied CDB-seq to RNA extracts, fresh frozen (FF) and formalin-fixed paraffin-embedded (FFPE) mouse brain tissue samples. CDB-seq transcriptomic profiles of tens of microdissected samples could be obtained in a simple, cost-effective way, providing a promising method for future spatial transcriptomics.
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Antiinfecciosos , Transcriptoma , Ratones , Animales , Transcriptoma/genética , Microesferas , Código de Barras del ADN Taxonómico , Fijación del Tejido/métodos , Perfilación de la Expresión Génica/métodos , ADN , FormaldehídoRESUMEN
Profiling gene expression while preserving cell locations aids in the comprehensive understanding of cell fates in multicellular organisms. However, simple and flexible isolation of microregions of interest (mROIs) for spatial transcriptomics is still challenging. We present a laser-induced forward transfer (LIFT)-based method combined with a full-length mRNA-sequencing protocol (LIFT-seq) for profiling region-specific tissues. LIFT-seq demonstrated that mROIs from two adjacent sections could reliably and sensitively detect and display gene expression. In addition, LIFT-seq can identify region-specific mROIs in the mouse cortex and hippocampus. Finally, LIFT-seq identified marker genes in different layers of the cortex with very similar expression patterns. These genes were then validated using in situ hybridization (ISH) results. Therefore, LIFT-seq will be a valuable and efficient technique for profiling the spatial transcriptome in various tissues.
Asunto(s)
Perfilación de la Expresión Génica , Transcriptoma , Animales , Perfilación de la Expresión Génica/métodos , Ratones , Rayos Láser , Hipocampo/metabolismo , ARN Mensajero/genética , ARN Mensajero/metabolismo , Hibridación in Situ/métodos , Corteza Cerebral/metabolismo , Análisis de Secuencia de ARN/métodosRESUMEN
Lesion-mimic mutants (LMM) spontaneously produce necrotic spots, a process not affected by environmental stress or pathogen infection. In this study, we identified a LMM, lesion mimic mutant 8 (lmm8) in rice (Oryza sativa). The lmm8 mutant produces brown and off-white lesions on its leaves during the second- and third-leaf stages. The lesion mimic phenotype of the lmm8 mutant was enhanced by light. At the mature stage, lmm8 mutant are shorter and exhibit inferior agronomic traits than the wild type. Contents of photosynthetic pigments and chloroplast fluorescence were significantly reduced in lmm8 leaves, along with increased production of reactive oxygen species and programmed cell death compared to the wild type. The mutated gene was identified as LMM8 (LOC_Os01g18320) by map-based cloning. A point mutation occurred in LMM8, causing a Leu to Arg mutation of the 146th amino acid of LMM8. It is an allele of SPRL1, encoding a protoporphyrinogen IX oxidase (PPOX) located in chloroplasts and involved in the biosynthesis of tetrapyrrole in chloroplasts. The lmm8 mutant showed enhanced resistance and broad-spectrum resistance. Together, our results demonstrate the importance of rice LMM8 protein in defense responses and plant growth in rice, and provides theoretical support for resistance breeding to improve rice yield.