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Tumor-associated hydrocephalus (TAH) is a common and lethal complication of brain metastases. Although other factors beyond mechanical obstructions have been suggested, the exact mechanisms are unknown. Using single-nucleus RNA sequencing and spatial transcriptomics, we find that a distinct population of mast cells locate in the choroid plexus and dramatically increase during TAH. Genetic fate tracing and intracranial mast-cell-specific tryptase knockout showed that choroid plexus mast cells (CPMCs) disrupt cilia of choroid plexus epithelia via the tryptase-PAR2-FoxJ1 pathway and consequently increase cerebrospinal fluid production. Mast cells are also found in the human choroid plexus. Levels of tryptase in cerebrospinal fluid are closely associated with clinical severity of TAH. BMS-262084, an inhibitor of tryptase, can cross the blood-brain barrier, inhibit TAH in vivo, and alleviate mast-cell-induced damage of epithelial cilia in a human pluripotent stem-cell-derived choroid plexus organoid model. Collectively, we uncover the function of CPMCs and provide an attractive therapy for TAH.
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Neoplasias Encefálicas , Plexo Corióideo , Hidrocefalia , Mastócitos , Humanos , Neoplasias Encefálicas/secundário , Plexo Corióideo/metabolismo , Plexo Corióideo/patologia , Hidrocefalia/metabolismo , Hidrocefalia/patologia , Mastócitos/metabolismo , Mastócitos/patologia , Triptases/líquido cefalorraquidiano , Metástase Neoplásica/patologiaRESUMO
Understanding molecular mechanisms that dictate B cell diversity is important for targeting B cells as anti-cancer treatment. Through the single-cell dissection of B cell heterogeneity in longitudinal samples of patients with breast cancer before and after neoadjuvant chemotherapy, we revealed that an ICOSL+ B cell subset emerges after chemotherapy. Using three immunocompetent mouse models, we recapitulated the subset switch of human tumor-infiltrating B cells during chemotherapy. By employing B-cell-specific deletion mice, we showed that ICOSL in B cells boosts anti-tumor immunity by enhancing the effector to regulatory T cell ratio. The signature of ICOSL+ B cells is imprinted by complement-CR2 signaling, which is triggered by immunogenic cell death. Moreover, we identified that CD55, a complement inhibitory protein, determines the opposite roles of B cells in chemotherapy. Collectively, we demonstrated a critical role of the B cell subset switch in chemotherapy response, which has implications in designing novel anti-cancer therapies. VIDEO ABSTRACT.
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Linfócitos B/imunologia , Neoplasias da Mama/imunologia , Ligante Coestimulador de Linfócitos T Induzíveis/metabolismo , Animais , Antineoplásicos/metabolismo , Linfócitos B/metabolismo , Antígenos CD55/imunologia , Antígenos CD55/metabolismo , Linhagem Celular Tumoral , Proteínas do Sistema Complemento/metabolismo , Modelos Animais de Doenças , Feminino , Humanos , Ligante Coestimulador de Linfócitos T Induzíveis/imunologia , Linfócitos do Interstício Tumoral/imunologia , Camundongos , Camundongos Endogâmicos C57BL , Receptores de Complemento 3d/imunologia , Receptores de Complemento 3d/metabolismo , Transdução de Sinais/imunologia , Linfócitos T Reguladores/imunologiaRESUMO
Reduced infiltration of anti-tumor lymphocytes remains a major cause of tumor immune evasion and is correlated with poor cancer survival. Here, we found that upregulation of regulator of G protein signaling (RGS)1 in helper TH1 cells and cytotoxic T lymphocytes (CTLs) reduced their trafficking to and survival in tumors and was associated with shorter survival of patients with breast and lung cancer. RGS1 was upregulated by type II interferon (IFN)-signal transducer and activator of transcription (STAT)1 signaling and impaired trafficking of circulating T cells to tumors by inhibiting calcium influx and suppressing activation of the kinases ERK and AKT. RGS1 knockdown in adoptively transferred tumor-specific CTLs significantly increased their infiltration and survival in breast and lung tumor grafts and effectively inhibited tumor growth in vivo, which was further improved when combined with programmed death ligand (PD-L)1 checkpoint inhibition. Our findings reveal RGS1 is important for tumor immune evasion and suggest that targeting RGS1 may provide a new strategy for tumor immunotherapy.
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Neoplasias da Mama/metabolismo , Carcinoma Ductal de Mama/metabolismo , Quimiotaxia de Leucócito , Linfócitos do Interstício Tumoral/metabolismo , Proteínas RGS/metabolismo , Subpopulações de Linfócitos T/metabolismo , Animais , Apoptose , Neoplasias da Mama/imunologia , Neoplasias da Mama/patologia , Neoplasias da Mama/terapia , Carcinoma Ductal de Mama/imunologia , Carcinoma Ductal de Mama/patologia , Carcinoma Ductal de Mama/terapia , Linhagem Celular Tumoral , Quimiocinas/metabolismo , Técnicas de Cocultura , Citotoxicidade Imunológica , Feminino , Humanos , Imunoterapia Adotiva , Linfócitos do Interstício Tumoral/imunologia , Linfócitos do Interstício Tumoral/transplante , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Microscopia de Fluorescência , Microscopia de Vídeo , Proteínas RGS/genética , Transdução de Sinais , Subpopulações de Linfócitos T/imunologia , Subpopulações de Linfócitos T/transplante , Linfócitos T Citotóxicos/imunologia , Linfócitos T Citotóxicos/metabolismo , Células Th1/imunologia , Células Th1/metabolismo , Fatores de Tempo , Imagem com Lapso de Tempo , Células Tumorais Cultivadas , Evasão TumoralRESUMO
Activation-induced cell death (AICD) of T lymphocytes can be exploited by cancers to escape immunological destruction. We demonstrated that tumor-specific cytotoxic T lymphocytes (CTLs) and type 1 helper T (TH1) cells, rather than type 2 helper T cells and regulatory T cells, were sensitive to AICD in breast and lung cancer microenvironments. NKILA, an NF-κB-interacting long noncoding RNA (lncRNA), regulates T cell sensitivity to AICD by inhibiting NF-κB activity. Mechanistically, calcium influx in stimulated T cells via T cell-receptor signaling activates calmodulin, thereby removing deacetylase from the NKILA promoter and enhancing STAT1-mediated transcription. Administering CTLs with NKILA knockdown effectively inhibited growth of breast cancer patient-derived xenografts in mice by increasing CTL infiltration. Clinically, NKILA overexpression in tumor-specific CTLs and TH1 cells correlated with their apoptosis and shorter patient survival. Our findings underscore the importance of lncRNAs in determining tumor-mediated T cell AICD and suggest that engineering lncRNAs in adoptively transferred T cells might provide a novel antitumor immunotherapy.
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Carcinoma/imunologia , RNA Longo não Codificante/imunologia , Linfócitos T Citotóxicos/imunologia , Células Th1/imunologia , Evasão Tumoral/genética , Animais , Apoptose/imunologia , Neoplasias da Mama/genética , Neoplasias da Mama/imunologia , Neoplasias da Mama/patologia , Carcinoma/genética , Carcinoma/patologia , Feminino , Xenoenxertos , Humanos , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/imunologia , Neoplasias Pulmonares/patologia , Masculino , Camundongos Endogâmicos NOD , Camundongos SCID , RNA Longo não Codificante/genéticaRESUMO
BACKGROUND: The efficacy of the BNT162b2 vaccine in pediatrics was assessed by randomized trials before the Omicron variant's emergence. The long-term durability of vaccine protection in this population during the Omicron period remains limited. OBJECTIVE: To assess the effectiveness of BNT162b2 in preventing infection and severe diseases with various strains of the SARS-CoV-2 virus in previously uninfected children and adolescents. DESIGN: Comparative effectiveness research accounting for underreported vaccination in 3 study cohorts: adolescents (12 to 20 years) during the Delta phase and children (5 to 11 years) and adolescents (12 to 20 years) during the Omicron phase. SETTING: A national collaboration of pediatric health systems (PEDSnet). PARTICIPANTS: 77 392 adolescents (45 007 vaccinated) during the Delta phase and 111 539 children (50 398 vaccinated) and 56 080 adolescents (21 180 vaccinated) during the Omicron phase. INTERVENTION: First dose of the BNT162b2 vaccine versus no receipt of COVID-19 vaccine. MEASUREMENTS: Outcomes of interest include documented infection, COVID-19 illness severity, admission to an intensive care unit (ICU), and cardiac complications. The effectiveness was reported as (1-relative risk)*100, with confounders balanced via propensity score stratification. RESULTS: During the Delta period, the estimated effectiveness of the BNT162b2 vaccine was 98.4% (95% CI, 98.1% to 98.7%) against documented infection among adolescents, with no statistically significant waning after receipt of the first dose. An analysis of cardiac complications did not suggest a statistically significant difference between vaccinated and unvaccinated groups. During the Omicron period, the effectiveness against documented infection among children was estimated to be 74.3% (CI, 72.2% to 76.2%). Higher levels of effectiveness were seen against moderate or severe COVID-19 (75.5% [CI, 69.0% to 81.0%]) and ICU admission with COVID-19 (84.9% [CI, 64.8% to 93.5%]). Among adolescents, the effectiveness against documented Omicron infection was 85.5% (CI, 83.8% to 87.1%), with 84.8% (CI, 77.3% to 89.9%) against moderate or severe COVID-19, and 91.5% (CI, 69.5% to 97.6%) against ICU admission with COVID-19. The effectiveness of the BNT162b2 vaccine against the Omicron variant declined 4 months after the first dose and then stabilized. The analysis showed a lower risk for cardiac complications in the vaccinated group during the Omicron variant period. LIMITATION: Observational study design and potentially undocumented infection. CONCLUSION: This study suggests that BNT162b2 was effective for various COVID-19-related outcomes in children and adolescents during the Delta and Omicron periods, and there is some evidence of waning effectiveness over time. PRIMARY FUNDING SOURCE: National Institutes of Health.
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Vacina BNT162 , COVID-19 , Estados Unidos , Humanos , Adolescente , Criança , Vacinas contra COVID-19 , COVID-19/prevenção & controle , Pesquisa Comparativa da Efetividade , HospitalizaçãoRESUMO
BACKGROUND: Eriophyoid mites (Eriophyoidea) are among the largest groups in the Acariformes; they are strictly phytophagous. The higher-level phylogeny of eriophyoid mites, however, remains unresolved due to the limited number of available morphological characters-some of them are homoplastic. Nevertheless, the eriophyoid mites sequenced to date showed highly variable mitochondrial (mt) gene orders, which could potentially be useful for resolving the higher-level phylogenetic relationships. RESULTS: Here, we sequenced and compared the complete mt genomes of 153 eriophyoid mite species, which showed 54 patterns of rearranged mt gene orders relative to that of the hypothetical ancestor of arthropods. The shared derived mt gene clusters support the monophyly of eriophyoid mites (Eriophyoidea) as a whole and the monophylies of six clades within Eriophyoidea. These monophyletic groups and their relationships were largely supported in the phylogenetic trees inferred from mt genome sequences as well. Our molecular dating results showed that Eriophyoidea originated in the Triassic and diversified in the Cretaceous, coinciding with the diversification of angiosperms. CONCLUSIONS: This study reveals multiple molecular synapomorphies (i.e. shared derived mt gene clusters) at different levels (i.e. family, subfamily or tribe level) from the complete mt genomes of 153 eriophyoid mite species. We demonstrated the use of derived mt gene clusters in unveiling the higher-level phylogeny of eriophyoid mites, and underlines the origin of these mites and their co-diversification with angiosperms.
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Genoma Mitocondrial , Magnoliopsida , Ácaros , Animais , Filogenia , Ácaros/genética , Genes Mitocondriais , Família Multigênica , Magnoliopsida/genéticaRESUMO
OBJECTIVES: It has become increasingly common for multiple computable phenotypes from electronic health records (EHR) to be developed for a given phenotype. However, EHR-based association studies often focus on a single phenotype. In this paper, we develop a method aiming to simultaneously make use of multiple EHR-derived phenotypes for reduction of bias due to phenotyping error and improved efficiency of phenotype/exposure associations. MATERIALS AND METHODS: The proposed method combines multiple algorithm-derived phenotypes with a small set of validated outcomes to reduce bias and improve estimation accuracy and efficiency. The performance of our method was evaluated through simulation studies and real-world application to an analysis of colon cancer recurrence using EHR data from Kaiser Permanente Washington. RESULTS: In settings where there was no single surrogate performing uniformly better than all others in terms of both sensitivity and specificity, our method achieved substantial bias reduction compared to using a single algorithm-derived phenotype. Our method also led to higher estimation efficiency by up to 30% compared to an estimator that used only one algorithm-derived phenotype. DISCUSSION: Simulation studies and application to real-world data demonstrated the effectiveness of our method in integrating multiple phenotypes, thereby enhancing bias reduction, statistical accuracy and efficiency. CONCLUSIONS: Our method combines information across multiple surrogates using a statistically efficient seemingly unrelated regression framework. Our method provides a robust alternative to single-surrogate-based bias correction, especially in contexts lacking information on which surrogate is superior.
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OBJECTIVE: To characterize the interplay between multiple medical conditions across sites and account for the heterogeneity in patient population characteristics across sites within a distributed research network, we develop a one-shot algorithm that can efficiently utilize summary-level data from various institutions. By applying our proposed algorithm to a large pediatric cohort across four national Children's hospitals, we replicated a recently published prospective cohort, the RISK study, and quantified the impact of the risk factors associated with the penetrating or stricturing behaviors of pediatric Crohn's disease (PCD). METHODS: In this study, we introduce the ODACoRH algorithm, a one-shot distributed algorithm designed for the competing risks model with heterogeneity. Our approach considers the variability in baseline hazard functions of multiple endpoints of interest across different sites. To accomplish this, we build a surrogate likelihood function by combining patient-level data from the local site with aggregated data from other external sites. We validated our method through extensive simulation studies and replication of the RISK study to investigate the impact of risk factors on the PCD for adolescents and children from four children's hospitals within the PEDSnet, A National Pediatric Learning Health System. To evaluate our ODACoRH algorithm, we compared results from the ODACoRH algorithms with those from meta-analysis as well as those derived from the pooled data. RESULTS: The ODACoRH algorithm had the smallest relative bias to the gold standard method (-0.2%), outperforming the meta-analysis method (-11.4%). In the PCD association study, the estimated subdistribution hazard ratios obtained through the ODACoRH algorithms are identical on par with the results derived from pooled data, which demonstrates the high reliability of our federated learning algorithms. From a clinical standpoint, the identified risk factors for PCD align well with the RISK study published in the Lancet in 2017 and other published studies, supporting the validity of our findings. CONCLUSION: With the ODACoRH algorithm, we demonstrate the capability of effectively integrating data from multiple sites in a decentralized data setting while accounting for between-site heterogeneity. Importantly, our study reveals several crucial clinical risk factors for PCD that merit further investigations.
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Algoritmos , Humanos , Criança , Adolescente , Reprodutibilidade dos Testes , Simulação por Computador , Modelos de Riscos Proporcionais , Funções VerossimilhançaRESUMO
While polymerization-induced self-assembly (PISA) has become a preferred synthetic route toward amphiphilic block copolymer self-assemblies, predicting their phase behavior from experimental design is extremely challenging, requiring time and work-intensive creation of empirical phase diagrams whenever self-assemblies of novel monomer pairs are sought for specific applications. To alleviate this burden, we develop here the first framework for a data-driven methodology for the probabilistic modeling of PISA morphologies based on a selection and suitable adaption of statistical machine learning methods. As the complexity of PISA precludes generating large volumes of training data with in silico simulations, we focus on interpretable low variance methods that can be interrogated for conformity with chemical intuition and that promise to work well with only 592 training data points which we curated from the PISA literature. We found that among the evaluated linear models, generalized additive models, and rule and tree ensembles, all but the linear models show a decent interpolation performance with around 0.2 estimated error rate and 1 bit expected cross entropy loss (surprisal) when predicting the mixture of morphologies formed from monomer pairs already encountered in the training data. When considering extrapolation to new monomer combinations, the model performance is weaker but the best model (random forest) still achieves highly nontrivial prediction performance (0.27 error rate, 1.6 bit surprisal), which renders it a good candidate to support the creation of empirical phase diagrams for new monomers and conditions. Indeed, we find in three case studies that, when used to actively learn phase diagrams, the model is able to select a smart set of experiments that lead to satisfactory phase diagrams after observing only relatively few data points (5-16) for the targeted conditions. The data set as well as all model training and evaluation codes are publicly available through the GitHub repository of the last author.
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Aprendizado de Máquina , Polimerização , Polímeros/química , Modelos LinearesRESUMO
BACKGROUND: Perfluoroheptanoic acid (PFHpA), a persistent organic pollutant widespread in the environment, is suspected as an environmental endocrine disruptor for its disturbance effect on hormone homeostasis and reproductive development. Whereas the effect of intrauterine PFHpA exposure during gestation on spermatogenesis of male offspring mice is still unknown. OBJECTIVE: This study aimed to explore the effect of prenatal PFHpA exposure on the reproductive development of male offspring mice and the role of N6-methyladenosine (m6A) during the process. METHODS: Fifty-six C57BL/6 pregnant mice were randomly divided into 4 groups. During the gestation period, the pregnant mice were exposed to 0, 0.0015, 0.015, and 0.15 mg/kg bw/d PFHpA from gestational day 1 (GD1) to GD16 by oral gavage. The male offspring mice were sacrificed by spinal dislocation at 7 weeks old. The body weight, testicular weight, and brain weight were weighed, and the intra-testicular testosterone was detected. The sperm qualities were analyzed with computer-aided sperm analysis (CASA). The testicular tissues were taken to analyze the pathological changes and examine the global m6A RNA methylation levels. Quantitative real-time PCR (qRT-PCR) was adopted to figure out the mRNA expression levels of m6A-related enzymes in testicular tissues of different PFHpA treated groups. Methylated RNA immunoprecipitation sequencing (MeRIP-seq) was applied to further explore the m6A RNA methylation at a whole-genome scale. RESULTS: Compared with the control group, no significant differences were observed in body weight, testicular weight, testicular coefficient, and the visceral-brain ratio of testicular tissue in the PFHpA treated groups. And no significant change was observed in intra-testicular testosterone among the four groups. CASA results showed a decrease of sperm count, sperm concentration, and total cell count, as well as an increase of sperm progressive cells' head area after prenatal PFHpA exposure (P < 0.05). Hematoxylin and eosin staining of pathological sections showed seminiferous tubules morphological change, disorder arrangement of seminiferous epithelium, and reduction of spermatogenic cells in the PFHpA treated groups. PFHpA significantly decreased global levels of m6A RNA methylation in testicular tissue (P < 0.05). Besides, qRT-PCR results showed significant alteration of the mRNA expression levels of seven m6A-related enzymes (Mettl3, Mettl5, Mettl14, Pcif1, Wtap, Hnrnpa2b1, and Hnrnpc) in the PFHpA treated groups (P < 0.05). MeRIP-seq results showed a correlation between prenatal PFHpA exposure and activation and binding of cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP). Cnga3 and Mpzl3 showed differential expression in the enrichment subcategories or pathways. CONCLUSIONS: Exposure to PFHpA during the gestation period would adversely affect the development of seminiferous tubules and testicular m6A RNA methylation in offspring mice, which subsequently interferes with spermatogenesis and leads to reproductive toxicity.
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Efeitos Tardios da Exposição Pré-Natal , Gravidez , Humanos , Feminino , Masculino , Animais , Camundongos , Efeitos Tardios da Exposição Pré-Natal/metabolismo , Camundongos Endogâmicos C57BL , Sêmen , Espermatogênese , Testículo , Testosterona/metabolismo , RNA Mensageiro/metabolismo , Peso Corporal , RNA/metabolismo , Proteínas de MembranaRESUMO
Microbial infections and microbial resistance lead to a high demand for new antimicrobial agents. Quaternized polysaccharides are cationic antimicrobial candidates; however, the limitation of homogeneous synthesis solvents that affect the molecular structure and biological activities, as well as their drug resistance remains unclear. Therefore, the authors homogeneously synthesize a series of quaternized chitin (QC) and quaternized chitosan (QCS) derivatives via a green and effective KOH/urea system and investigate their structure-activity relationship and biological activity in vivo and in vitro. Their study reveals that a proper match of degree of quaternization (DQ) and degree of deacetylation (DD') of QC or QCS is key to balance antimicrobial property and cytotoxicity. They identify QCS-2 as the optimized antimicrobial agent with a DQ of 0.46 and DD' of 82%, which exhibits effective broad-spectrum antimicrobial properties, good hemocompatibility, excellent cytocompatibility, and effective inhibition of bacterial biofilm formation and eradication of mature bacterial biofilms. Moreover, QCS-2 exhibits a low propensity for development of drug resistance and significant anti-infective effects on MRSA in vivo comparable to that of vancomycin, avoiding excessive inflammation and promoting the formation of new blood vessels, hair follicles, and collagen deposition to thus expedite wound healing.
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Anti-Infecciosos , Infecções Bacterianas , Quitosana , Antibacterianos/química , Antibacterianos/farmacologia , Anti-Infecciosos/farmacologia , Biofilmes , Quitosana/química , Farmacorresistência Bacteriana , Humanos , Micelas , Testes de Sensibilidade MicrobianaRESUMO
In this work, we present a coherent distributed radio frequency (RF) array, discover and quantitatively describe the strong positive correlation between reconstructed signals for the first time. Eight replicable parallel receivers are connected to the phase-locked common trunk link via eight optical couplers spaced 1 km apart. The forward and backward signals at each receiver, extracted from two ports of optical couplers, are recovered to RF signals separately and then mixed to achieve upward frequency conversion. The link delay jitter is counteracted by wavelength-tuning of the optical carrier. With the long-term stability of point-to-multipoint fiber-optic RF dissemination effectively improved, the coherent distributed array is generated, and further the relative frequency stability between signals at different receivers is studied. The proposed correlation coefficient at 103 s is â¼0.8 and shows a slight downward trend with the increase of averaging time based on our experimental results.
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BACKGROUND AND AIMS: Effective traction is an important prerequisite for successful endoscopic submucosal dissection (ESD). The combined orthodontic rubber band (ORB) and clip method was effective in colorectal cancer ESD. To date, the method was not reported in gastric ESD. This study aimed to investigate its efficacy and safety for gastric neoplasms ESD. METHODS: We retrospectively analyzed data of 118 patients with gastric neoplasms treated by ESD from November 2020 to April 2022, 43 by ORB-ESD and 75 by the conventional ESD. The primary outcome measure was the ESD procedure time. Clinical data on efficacy and safety were also collected and analyzed. Propensity score matching (PSM) matched the patients in both groups. RESULTS: PSM successfully matched 31 pairs of patients. The ORB-ESD operation time was shorter (median [interquartile range], 35 [30-48] vs. 49 [40-70] min, P < 0.001) and dissection speed was higher (median [interquartile range], 22.6 [14.4-29.3] vs. 13.5 [9.6-17.9] mm2/min, P < 0.001) than in the conventional ESD. The groups were similar in muscular injury rate, frequency and time of use of thermal hemostatic forceps, postoperative adverse events, en bloc resection, and R0 resection rate (P > 0.05). CONCLUSIONS: Compared to the conventional ESD, ORB-ESD significantly reduced the procedure time and increased the dissection speed, proving beneficial to gastric ESD.
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Ressecção Endoscópica de Mucosa , Neoplasias Gástricas , Humanos , Ressecção Endoscópica de Mucosa/efeitos adversos , Ressecção Endoscópica de Mucosa/métodos , Neoplasias Gástricas/cirurgia , Neoplasias Gástricas/etiologia , Estudos Retrospectivos , Instrumentos Cirúrgicos , Dissecação/efeitos adversos , Dissecação/métodos , Resultado do TratamentoRESUMO
Alfalfa (Medicago sativa) is the most important perennial forage legume worldwide, with high yields and nutrient quality. Diseases are significant threats to alfalfa and cause substantial yield losses (Fang et al. 2021). In March 2022, a disease with symptoms similar to fusarium wilt was observed in an alfalfa (cultivar Aurora) field in Baima teaching and research base (119°18'07â³E, 31°61'47â³N) in Nanjing, China. Reddish-brown discoloration of the roots, stele, basal stems and the withering symptoms on the aerial portions are the specific symptoms of fusarium wilt. The disease incidence varied from 3 to 6% in around 0.3 hectares of alfalfa fields surveyed. Fresh tissues from symptomatic alfalfa were cut into pieces (3-5 mm) and surface sterilized with 75% ethanol for 30 seconds, followed by 1% sodium hypochlorite (NaClO) for 120 seconds, and three rinses with sterile distilled water. Tissue pieces were placed on selective potato dextrose agar (PDA) containing 50 mg/L rifampicin and ampicillin. Plates were sealed and incubated at 25°C for 48 to 72 hours. Over 70 tissue fragments plated, 20 isolates from different fragments showed similar colonies. These isolates were purified by single spore isolation and grown on PDA and mung bean medium for morphology identification, molecular identification and pathogenicity test. The colony showed white to pink, abundant, densely aerial mycelium, while its backside was light violet. Macroconidia were hyaline, falcate with septa, and ranged in size from 25.5 to 61.5 × 3.8 to 6.2 µm (n=50). Microconidia were non-septate, hyaline, oval, straight to slightly curved, and were 5.6 to 10.7 × 2.4 to 3.2 µm (n=50). Spherical chlamydospores were 10.6 ± 1.3 µm in size. The rDNA internal transcribed spacers (ITS), RNA polymerase II subunit 2 (RPB2) and translation elongation factor 1-α (TEF) genes were amplified and sequenced (White et al. 1990; Carbone et al. 1999; Wang et al. 2019). Blast analysis showed 99.41% to 100% identity to F. commune sequences in the GenBank database (KU341324.1, MN892350.1, MH341219.1). The pathogenicity test was conducted using the dipping method in roots. Fresh F. commune hyphae were cut into 3 x 3mm agar plugs from a 7 cm PDA plate and inoculated in 200 mL mung bean medium on a shaker at 160 rpm, 25°C for 5 days. Spores were filtered through a cheese cloth, adjusted to 1 × 106 spores/mL with distilled water and used immediately. Roots in two-week-old alfalfa seedlings were soaked in spore suspension for 30 mins before being transplanted to sterile vermiculate with four repeats, while those in sterile water were used as non-inoculated control. The plants were placed in the green house at 25°C with 16 h of light and 8 h of darkness. The symptoms were observed two weeks following the inoculation. 30% of the seedlings withered, and reddish-brown discoloration symptoms were visible on the roots and stem of the unwithered plant. F. commune was isolated from these lesions again, and no disease symptoms were observed in the control plant. F. commune has been reported to cause root and stem rot in many plants worldwide, including tobacco (Zhong et al. 2021), maize (Mezzalama et al. 2021; Xi et al. 2019), soybean (Detranaltes et al. 2022) and sugarcane (Wang et al. 2018). Our results show that F. commune was the causative agent of alfalfa root rot. To our knowledge, this is the first report of F. commune causing root rot in alfalfa. The finding provides insights for disease diagnosis of alfalfa root rot disease and management in the region.
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Although CD4+ CD45RA- Foxp3l ° cytokine-secreting T cells (Fr.III cells) have been reported to be increased in systemic lupus erythematosus (SLE), their function and effects on response of B cells are still unclear. Here, we dissect how BACH2 regulates Fr.III cells function and promotes B-cell response in active SLE patients. We measured cytokines and BACH2 expression, and found that Fr.III cells from SLE patients produce much more inflammatory cytokines and were more able to promote B- cell proliferation, IgG, IgA, and TNF-α production than controls in a co-culture system. Fr.III cells expressed high levels of ICOS and CD154, but a low level of Tfr and BACH2, BACH2 expression was negatively correlated with SLE Disease Activity Index. Overexpressed of BACH2 in Fr.III cells, decreased cytokines expression and reduced B-cell response. Furthermore, we identified a reduction of H3K27ac level binding at the BACH2 locus in the SLE Fr.III cells and SLE serum stimulation decreased H3K27ac binding at the BACH2 locus, which could be restored using trichostatin A (TSA). In conclusion, BACH2 was associated with SLE disease activity, regulated the function of Fr.III cells, and promoted B-cells response. Targeting BACH2 may be a new immune intervention therapy of SLE.
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Linfócitos B/imunologia , Fatores de Transcrição de Zíper de Leucina Básica/imunologia , Lúpus Eritematoso Sistêmico/imunologia , Subpopulações de Linfócitos T/imunologia , Adulto , Feminino , Humanos , Ativação Linfocitária/imunologia , Masculino , Pessoa de Meia-IdadeRESUMO
BACKGROUND: This systematic review and meta-analysis aimed to investigate local recurrence (LR) rates among the three grades (benign, borderline, and malignant) of phyllodes tumors (PTs). The study also assessed various risk factors for LR. METHODS: Electronic articles published between 1 January 1995 and 31 May 2018, were searched and critically appraised. The authors independently reviewed the abstracts and extracted data for LR rates and LR risk factors. RESULTS: The review incorporated 54 studies with 9234 individual cases. The pooled LR rates were 8% for benign, 13% for borderline, and 18% for malignant PTs. The risk of LR was significantly increased by borderline versus benign PTs (odds ratio [OR] 2.00; 95% confidence interval [CI] 1.68-2.38) and malignant versus borderline PTs (OR 1.28; 95% CI 1.05-1.55). The significant risk factors for LR were mitoses, tumor border (infiltrating vs. pushing), stromal cellularity (moderate/severe vs. mild), stromal atypia (severe vs. mild/absent), stromal overgrowth (severe vs. mild/absent), and tumor necrosis (positive vs. negative). Age and tumor size were not associated with LR risk. The subgroup analysis showed that breast-conserving surgery versus mastectomy and positive versus negative surgical margins were significantly associated with an increased LR risk only in malignant PTs. CONCLUSIONS: The risk of LR was significantly increased from benign to borderline to malignant PTs. Mitoses, tumor border, stromal cellularity, stromal atypia, stromal overgrowth, tumor necrosis, type of surgery, and surgical margin status may be risk factors for LR. Different management strategies could be considered for different PT grades.
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Neoplasias da Mama/patologia , Recidiva Local de Neoplasia/patologia , Tumor Filoide/patologia , Neoplasias da Mama/complicações , Feminino , Humanos , Recidiva Local de Neoplasia/etiologia , Tumor Filoide/complicações , Fatores de RiscoRESUMO
CONTEXT: Osteoporosis (OP) is a progressive systemic bone disease. Dual-energy X-ray absorptiometry (DXA) is routinely employed and is considered the gold standard method for the diagnosis of OP. OBJECTIVE: We aimed to investigate the potential use of combined information from multiple bone turnover markers (BTMs) as a clinical diagnostic tool for OP. MATERIALS AND METHODS: A total of 9053 Chinese postmenopausal women (2464 primary OP patients and 6589 healthy controls) were recruited. Serum levels of six common BTMs, including BAP, BSP, CTX, OPG, OST and sRANKL were assayed. Models based on support vector machine (SVM) were constructed to explore the efficiency of different combinations of multiple BTMs for OP diagnosis. RESULTS: Increasing the number of BTMs used in generating the models increased the predictive power of the SVM models for determining the disease status of study subjects. The highest kappa coefficient for the model with one BTM (BAP) compared to DXA was 0.7783. The full model incorporating all six BTMs resulted in a high kappa coefficient of 0.9786. CONCLUSION: Our findings showed that although single BTMs were not sufficient for OP diagnosis, appropriate combinations of multiple BTMs incorporated into the SVM models showed almost perfect agreement with the DXA.
Assuntos
Biomarcadores/sangue , Remodelação Óssea/genética , Osteoporose/sangue , Absorciometria de Fóton , Idoso , Densidade Óssea/genética , China/epidemiologia , Enzimas de Restrição do DNA/sangue , Feminino , Fatores de Troca do Nucleotídeo Guanina/sangue , Humanos , Pessoa de Meia-Idade , Osteoporose/diagnóstico por imagem , Osteoporose/patologia , Ligante RANK/sangue , Fatores de Troca de Nucleotídeo Guanina Rho/sangue , Máquina de Vetores de SuporteRESUMO
Objective: To investigate the benefits and drawbacks of breast reconstruction with endoscopic-assisted harvesting of the latissimus dorsi muscle flap for breast cancer and treatment experience of postoperative operation-related complications. Methods: A retrospective analysis was performed on clinical data of 26 female patients with breast cancer who met the selection criteria between September 2021 and March 2023 aging 48.7 years (range, 26-69 years). All tumors were unilateral, with 17 on the left side and 9 on the right side. The tumor size ranged from 1.0 to 7.0 cm, with an average of 2.7 cm. The pathological staging included T 1 in 11 cases, T 2 in 14 cases, and T 3 in 1 case; N 0 in 10 cases, N 1 in 11 cases, N 2 in 2 cases, and N 3 in 3 cases; no distant metastasis (M 0) occurred when first diagnosed. Among them, 10 cases underwent breast conserving surgery, and 16 cases underwent nipple-sparing mastectomy. All patients underwent breast reconstruction with endoscopic-assisted harvesting of the latissimus dorsi muscle flap. The operation time, incision length, and postoperative drainage volume in 3 days were recorded. Breast-Q "Satisfaction with back" scale was conducted to evaluate patients' satisfaction with back at 6 months after operation. Results: The operation time was 280-480 minutes (mean, 376.7 minutes), the incision length was 10-15 cm (mean, 12.2 cm), the postoperative drainage volume in 3 days was 500-1 600 mL (mean, 930.2 mL). There were 4 cases of postoperative seroma, 1 case of incision rupture, 1 case of paresthesia of the thoracic wall, and 1 case of edema of the ipsilateral upper limb. All patients were followed up 12-30 months (mean, 20.1 months). No latissimus dorsi muscle flap necrosis, latissimus dorsi muscle atrophy, or shoulder joint dysfunction occurred during follow-up; 2 patients had recurrence of lymph nodes in the ipsilateral axilla after operation, but no distant metastasis occurred. Breast-Q score at 6 months after operation was 64-100 (mean, 79.5). The average score was 78.6 (range, 64-100) in patients underwent nipple-sparing mastectomy and 81.0 (range, 78-100) in patients underwent breast conserving surgery. Conclusion: Breast reconstruction with endoscopic-assisted harvesting of the latissimus dorsi muscle flap for breast cancer is proven to be a surgical approach with safety and cosmetic effects with mild postoperative operation-related complications and considerable patient satisfaction.
Assuntos
Neoplasias da Mama , Endoscopia , Mamoplastia , Complicações Pós-Operatórias , Músculos Superficiais do Dorso , Retalhos Cirúrgicos , Humanos , Feminino , Pessoa de Meia-Idade , Neoplasias da Mama/cirurgia , Neoplasias da Mama/patologia , Mamoplastia/métodos , Adulto , Estudos Retrospectivos , Músculos Superficiais do Dorso/transplante , Endoscopia/métodos , Idoso , Mastectomia/métodos , Mastectomia Segmentar/métodosRESUMO
Collective cell migration is an important process in cancer metastasis. Unlike single-cell migration, collective cell migration requires E-cadherin expression in the cell cohort. However, the mechanisms underlying cellular contact and focal adhesions remain unclear. In this study, Src was hypothesized to coordinate focal adhesion and Rab11-mediated E-cadherin distribution during collective cell migration. This study primarily used confocal microscopy to visualize the 3D structure of cell-cell contacts with associated molecules. These results demonstrate that the clinical Src inhibitor dasatinib was less toxic to HT-29 colon cancer cells; instead, the cells aggregated. 3D immunofluorescence imaging showed that Rab11 was localized with E-cadherin at the adherens junctions of the apical cell-cell contacts. In the transwell assay, Rab11 colocalized with a broad range of E-cadherin proteins in collectively migrated cells, and dasatinib treatment significantly suppressed collective cell migration. Transmission electron microscopy demonstrated that dasatinib treatment increased cell membrane protrusion contacts and generated spaces between cells, which may allow epidermal growth factor receptor activity at the cell-cell contacts. This study suggests that dasatinib treatment does not inhibit cell survival but targets Src at different cellular compartments in the coordination of focal adhesions and cell-cell contacts in collective cell migration through E-cadherin dynamics in colon cancer cells.
RESUMO
Hypoxanthine phosphoribosyl transferase 1 (HPRT1), once considered a housekeeping gene, has been identified as playing an important role in several tumors. Its role in pan-cancer, however, has not been systematically studied. This study evaluates the relationship between HPRT1 and clinical parameters, survival prognosis, and tumor immunity based on multi omics data from the Cancer Genome Atlas (TCGA) and the Gene Expression Omnibus (GEO) databases. Drug sensitivity analysis screened 16 effective drugs against HPRT1, exploring the interactions with chemicals and genes. The significance of HPRT1 in tumor immunotherapy has also been investigated. Immunohistochemistry confirmed significant differences in the expression of HPRT1 between five tumor types (colon adenocarcinoma [COAD], head-neck squamous cell carcinoma [HNSC], lung adenocarcinoma [LUAD], thyroid carcinoma [THCA], and uterine corpus endometrial carcinoma [UCEC]) and adjacent normal tissues (P < 0.05). HPRT1 competitive endogenous RNA network was constructed in HNSC. Through cytological experiments, it was verified that HPRT1 plays a carcinogenic role in HNSC and is associated with tumor cell proliferation, migration, invasion, and apoptosis. In addition, there was a significant positive correlation between HPRT1 and programmed cell death-1 (PD-1) expression in HNSC (P < 0.05). These findings suggest that HPRT1 may be a potential biomarker for predicting and treating cancer.