RESUMO
Lymph node status is a key factor in determining stage, treatment, and prognosis in cancers. Small lymph nodes in the fat-rich gastrointestinal and breast cancer specimens are easily missed in conventional sampling methods. This study examined the effectiveness of degreasing pretreatment with dimethyl sulfoxide (DMSO) in lymph node detection and its impact on the analysis of clinical treatment-related proteins and molecules. Thirty-three cases of gastrointestinal cancer specimens from radical gastrectomy and 63 cases of breast cancer specimens from modified radical mastectomy were included. After routine sampling of lymph nodes, the specimens were immersed in DMSO for 30 minutes for defatting. We assessed changes in the number of detected lymph nodes and pN staging in 33 gastrointestinal cancer specimens and 37 breast cancer specimens. Additionally, we analyzed histological characteristics, Masson's trichrome special staining, and immunohistochemistry (gastrointestinal cancer: MMR, HER2, PD-L1; breast cancer: ER, PR, AR, HER2, Ki-67, PD-L1). Molecular status was evaluated for colorectal cancer (KRAS, NRAS, BRAF, MSI) and breast cancer (HER2) in gastrointestinal cancer specimens and the remaining 26 breast cancer specimens. Compared to conventional sampling, DMSO pretreatment increased the detection rate of small lymph nodes (gastrointestinal cancer: p<0.001; breast cancer: p<0.001) and improved pN staging in one case each of gastric cancer, colon cancer, and rectal cancer (3/33, 9.1%). No significant difference in the morphology, special staining, protein, and molecular status of cancer tissue after DMSO treatment were found. Based on these results and our institutional experience, we recommend incorporating DMSO degreasing pretreatment into clinical pathological sampling practices.
RESUMO
Consumption of dietary fiber and anthocyanin has been linked to a lower incidence of colorectal cancer (CRC). This study scrutinizes the potential antitumorigenic attributes of a black rice diet (BRD), abundantly rich in dietary fiber and anthocyanin. Our results demonstrate notable antitumorigenic effects in mice on BRD, indicated by a reduction in both the size and number of intestinal tumors and a consequent extension in life span, compared to control diet-fed counterparts. Furthermore, fecal transplants from BRD-fed mice to germ-free mice led to a decrease in colonic cell proliferation, coupled with maintained integrity of the intestinal barrier. The BRD was associated with significant shifts in gut microbiota composition, specifically an augmentation in probiotic strains Bacteroides uniformis and Lactobacillus. Noteworthy changes in gut metabolites were also documented, including the upregulation of indole-3-lactic acid and indole. These metabolites have been identified to stimulate the intestinal aryl hydrocarbon receptor pathway, inhibiting CRC cell proliferation and colorectal tumorigenesis. In summary, these findings propose that a BRD may modulate the progression of intestinal tumors by fostering protective gut microbiota and metabolite profiles. The study accentuates the potential health advantages of whole-grain foods, emphasizing the potential utility of black rice in promoting health.
RESUMO
Technology for spatial multi-omics aids the discovery of new insights into cellular functions and disease mechanisms. Here we report the development and applicability of multi-omics in situ pairwise sequencing (MiP-seq), a method for the simultaneous detection of DNAs, RNAs, proteins and biomolecules at subcellular resolution. Compared with other in situ sequencing methods, MiP-seq enhances decoding capacity and reduces sequencing and imaging costs while maintaining the efficacy of detection of gene mutations, allele-specific expression and RNA modifications. MiP-seq can be integrated with in vivo calcium imaging and Raman imaging, which enabled us to generate a spatial multi-omics atlas of mouse brain tissues and to correlate gene expression with neuronal activity and cellular biochemical fingerprints. We also report a sequential dilution strategy for resolving optically crowded signals during in situ sequencing. High-throughput in situ pairwise sequencing may facilitate the multidimensional analysis of molecular and functional maps of tissues.
Assuntos
Sequenciamento de Nucleotídeos em Larga Escala , Animais , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Camundongos , Encéfalo/metabolismo , Cálcio/metabolismo , Genômica/métodos , MultiômicaRESUMO
Endolymphatic sac tumor (ELST) is a rare disease that originates from the endolymphatic sac system of the inner ear. Being a low-grade malignant tumor, ELST has a mild morphology and is characterized by a slow but aggressive growth. Most clinicians and pathologists are unfamiliar with this disease. ELST can be misdiagnosed as metastatic renal cancer because of the similarity in morphology and expression of nephrogenic markers such as PAX8. The presented case of a 27-year-old man revealed that observing the characteristic location and confirming the absence of renal neoplasm to rule out the possibility of metastasis are critical for obtaining an accurate final diagnosis.
Assuntos
Adenoma , Neoplasias Ósseas , Carcinoma de Células Renais , Neoplasias da Orelha , Saco Endolinfático , Neoplasias Renais , Masculino , Humanos , Adulto , Carcinoma de Células Renais/diagnóstico , Neoplasias Renais/diagnóstico , Neoplasias Renais/patologia , Saco Endolinfático/química , Saco Endolinfático/patologia , Imuno-Histoquímica , Neoplasias da Orelha/diagnóstico , Neoplasias da Orelha/química , Neoplasias da Orelha/patologia , Neoplasias Ósseas/patologia , Adenoma/patologia , Erros de DiagnósticoRESUMO
Single-cell and spatial transcriptome sequencing, two recently optimized transcriptome sequencing methods, are increasingly used to study cancer and related diseases. Cell annotation, particularly for malignant cell annotation, is essential and crucial for in-depth analyses in these studies. However, current algorithms lack accuracy and generalization, making it difficult to consistently and rapidly infer malignant cells from pan-cancer data. To address this issue, we present Cancer-Finder, a domain generalization-based deep-learning algorithm that can rapidly identify malignant cells in single-cell data with an average accuracy of 95.16%. More importantly, by replacing the single-cell training data with spatial transcriptomic datasets, Cancer-Finder can accurately identify malignant spots on spatial slides. Applying Cancer-Finder to 5 clear cell renal cell carcinoma spatial transcriptomic samples, Cancer-Finder demonstrates a good ability to identify malignant spots and identifies a gene signature consisting of 10 genes that are significantly co-localized and enriched at the tumor-normal interface and have a strong correlation with the prognosis of clear cell renal cell carcinoma patients. In conclusion, Cancer-Finder is an efficient and extensible tool for malignant cell annotation.
Assuntos
Carcinoma de Células Renais , Neoplasias Renais , Humanos , Carcinoma de Células Renais/genética , Perfilação da Expressão Gênica , Transcriptoma/genética , Algoritmos , Neoplasias Renais/genética , Análise de Célula ÚnicaRESUMO
Syncytial giant cells (SGCs) are neoplastic giant cells of epithelial origin. The nuclear morphology of SGCs is uniform and similar to those of adjacent mononuclear tumor cells. Clear cell renal cell carcinoma (ccRCC) with SGCs is a rare microscopic morphology. In this study, the clinical and pathological data of 16 ccRCC cases with SGCs were retrospectively reviewed. The incidence of SGCs in pathological stages pT3 and above (12.1%, 8/66) was significantly higher than that in pT1 and pT2 (2.6%, 8/306) (P = 0.002). The incidence of SGCs in the WHO/ISUP nuclear grade 3 or 4 ccRCC (12.4%, 14/113) was significantly higher than that in grade 1 or 2 (0.8%, 2/259) (P < 0.001). Two forms of SGCs were observed, some exhibited nuclear pyknosis and degeneration. Of the 16 cases, eight cases were accompanied by necrosis and seven cases had lymphovascular invasion. Both SGCs and mononuclear tumor cells were positive for ccRCC markers (PAX8, CAIX, CD10 and Vimentin). None of the SGC nuclei were positive for Ki-67. Follow-up information was available on 14 patients, with a median follow-up time of 27.5 months. Ten patients were alive without disease, three were alive with metastatic disease, and one patient died 10 months after surgery. These findings indicated that SGCs are not rare, especially in ccRCC with high nuclear grade and pathological stage, and often co-exist with other adverse prognostic features. SGCs may be senescent tumor cells, the presence of SGCs should not be considered as Fuhrman and WHO/ISUP nuclear grading 4.
Assuntos
Carcinoma de Células Renais , Neoplasias Renais , Humanos , Carcinoma de Células Renais/patologia , Neoplasias Renais/patologia , Estudos Retrospectivos , Prognóstico , Células Gigantes/patologiaRESUMO
ISGylation is a well-established antiviral mechanism, but its specific function in immune and tissue homeostasis regulation remains elusive. Here, we reveal that the RNA-binding protein RBM47 undergoes phosphorylation-dependent ISGylation at lysine 329 to regulate immune activation and maintain lung homeostasis. K329R knockin (KI) mice with defective RBM47-ISGylation display heightened susceptibility to LPS-induced acute lung injury and lung tumorigenesis, accompanied with multifaceted immunosuppression characterized by elevated pro-inflammatory factors, reduced IFNs/related chemokines, increased myeloid-derived suppressor cells, and impaired tertiary lymphoid structures. Mechanistically, RBM47-ISGylation regulation of the expression of TSC22D3 mRNA, a glucocorticoid-inducible transcription factor, partially accounts for the effects of RBM47-ISGylation deficiency due to its broad immunosuppressive activity. We further demonstrate the direct inhibitory effect of RBM47-ISGylation on TSC22D3 expression in human cells using a nanobody-targeted E3 ligase to induce site-specific ISGylation. Furthermore, epinephrine-induced S309 phosphorylation primes RBM47-ISGylation, with epinephrine treatment exacerbating dysregulated cytokine expression and ALI induction in K329R KI mice. Our findings provide mechanistic insights into the dynamic regulation of RBM47-ISGylation in supporting immune activation and maintaining lung homeostasis.
RESUMO
Men demonstrate higher incidence and mortality rates of colorectal cancer (CRC) than women. This study aims to explain the potential causes of such sexual dimorphism in CRC from the perspective of sex-biased gut microbiota and metabolites. The results show that sexual dimorphism in colorectal tumorigenesis is observed in both ApcMin/ + mice and azoxymethane (AOM)/dextran sulfate sodium (DSS)-treated mice with male mice have significantly larger and more tumors, accompanied by more impaired gut barrier function. Moreover, pseudo-germ mice receiving fecal samples from male mice or patients show more severe intestinal barrier damage and higher level of inflammation. A significant change in gut microbiota composition is found with increased pathogenic bacteria Akkermansia muciniphila and deplets probiotic Parabacteroides goldsteinii in both male mice and pseudo-germ mice receiving fecal sample from male mice. Sex-biased gut metabolites in pseudo-germ mice receiving fecal sample from CRC patients or CRC mice contribute to sex dimorphism in CRC tumorigenesis through glycerophospholipids metabolism pathway. Sexual dimorphism in tumorigenesis of CRC mouse models. In conclusion, the sex-biased gut microbiome and metabolites contribute to sexual dimorphism in CRC. Modulating sex-biased gut microbiota and metabolites could be a potential sex-targeting therapeutic strategy of CRC.
Assuntos
Neoplasias Colorretais , Microbioma Gastrointestinal , Masculino , Feminino , Animais , Camundongos , Neoplasias Colorretais/patologia , Sulfato de Dextrana , Carcinogênese , Transformação Celular NeoplásicaRESUMO
OBJECTIVE: We report our diagnosis of a rare case of primary angiomatoid fibrous histiocytoma in the mandible of a 42-year-old male using next-generation sequencing to detect disease-specific EWSR1-ATF1 fusion. STUDY DESIGN: After the initial cone beam computerized tomography scan and reconstruction, we performed immunohistochemical staining and fluorescence in situ hybridization analysis on tissue samples to detect EWSR1 gene rearrangement. For the final diagnosis, we performed next-generation sequencing to detect disease-specific EWSR1-ATF1 fusion. RESULTS: FISH analysis showed approximately 55% of tumor cells with mostly isolated red signals, as well as several split red-green signals, indicating the presence of EWSR1 gene rearrangement. Next-generation sequencing analysis identified an EWSR1 exon9-ATF1 exon4 fusion, a diagnostic biomarker of angiomatoid fibrous histiocytoma (AFH). Based on the findings, we diagnosed primary AFH derived from the mandible. CONCLUSIONS: Next-generation sequencing is a powerful methodology for detecting disease-specific EWSR1-ATF1 fusion and diagnosing primary angiomatoid fibrous histiocytoma.
Assuntos
Histiocitoma Fibroso Benigno , Histiocitoma Fibroso Maligno , Masculino , Humanos , Hibridização in Situ Fluorescente , Proteína EWS de Ligação a RNA/genética , Histiocitoma Fibroso Maligno/diagnóstico por imagem , Histiocitoma Fibroso Maligno/genética , Tomografia Computadorizada de Feixe Cônico , Proteínas de Fusão Oncogênica/genética , Proteínas de Fusão Oncogênica/metabolismoRESUMO
Surgery-induced renal ischemia and reperfusion (I/R) injury and nephrotoxic drugs like cisplatin can cause acute kidney injury (AKI), for which there is no effective therapy. Lipid accumulation is evident following AKI in renal tubules although the mechanisms and pathological effects are unclear. Here, we report that Ehmt2-encoded histone methyltransferase G9a is upregulated in patients and mouse kidneys after AKI. Renal tubular specific knockout of G9a (Ehmt2Ksp ) or pharmacological inhibition of G9a alleviates lipid accumulation associated with AKI. Mechanistically, G9a suppresses transcription of the lipolytic enzyme Ces1; moreover, G9a and farnesoid X receptor (FXR) competitively bind to the same promoter regions of Ces1. Ces1 is consistently observed to be downregulated in the kidney of AKI patients. Pharmacological inhibition of Ces1 increases lipid accumulation, exacerbates renal I/R-injury and eliminates the beneficial effects on AKI observed in Ehmt2Ksp mice. Furthermore, lipid-lowering atorvastatin and an FXR agonist alleviate AKI by activating Ces1 and reducing renal lipid accumulation. Together, our results reveal a G9a/FXR-Ces1 axis that affects the AKI outcome via regulating renal lipid accumulation.
Assuntos
Injúria Renal Aguda , Túbulos Renais , Camundongos , Animais , Túbulos Renais/metabolismo , Túbulos Renais/patologia , Injúria Renal Aguda/genética , Injúria Renal Aguda/induzido quimicamente , Lipídeos , Rim/patologia , Camundongos Endogâmicos C57BLRESUMO
Background: Pure apocrine carcinoma (AC) of the breast can be divided into human epidermal growth factor receptor-2 (HER2)-positive and triple-negative apocrine carcinoma (TNAC). Some studies showed that triple negative breast cancer with low tumor-infiltrating lymphocytes (TILs) and high programmed death-ligand 1 (PD-L1) status may be a therapeutic target for immune checkpoint inhibitors. However, the clinicopathological features of different HER2 expression, TILs status and PD-L1 expression in AC are not clear. Therefore, we investigate the status of TILs and PD-L1, as well as the clinicopathological features of pure apocrine carcinoma of the breast. Methods: We retrospectively analyzed the clinicopathological data, and prognosis of 41 cases of pure apocrine carcinoma of the breast that underwent surgical resection from January 2014 to November 2020. TILs were evaluated. Immunohistochemistry (IHC) staining was applied to detect PD-L1 protein expression in 14 of these samples from January 2019 to November 2020. The expression and correlation of HER2, TILs, PD-L1 and clinicopathological features and prognoses were analyzed. Results: A total of 80.5% (33/41) of patients had TILs <50%, and 19.5% (8/41) had TILs ≥50%. The expression of TILs and the Ki-67 proliferation index were significantly higher in the HER2-positive group (41.5%, 17/41) compared to the HER2-negative group (58.5%, 24/41) (P<0.05). Approximately 52.9% (9/17) of HER2-positive patients treated with Trastuzumab targeted therapy, overall survival was higher in HER2-positive patients than in HER2-negative patients (P=0.211). The PD-L1 positivity rate was 50% (7/14) in the 14 pure apocrine carcinoma of the breast samples, and 66.7% (4/6) and 37.5% (3/8) in the HER2-negative and HER2-positive groups, respectively, with no significant difference (P=0.592). Among these 14 cases, two samples had TILs ≥50%, both of which were positive for PD-L1 and Ki67 >20%; and 12 cases had TILs <50%, of which 41.7% (5/12) were PD-L1-positive and 58.3% (7/12) were PD-L1-negative. All 14 cases with PD-L1-negative had TILs <50%. There was no significant difference in overall survival between TILs and Ki67 co-expression (P=0.452). Conclusions: Pure AC HER2-positive patients have higher levels of TILs and Ki67, HER2 negative and TILs ≥50% patients may have higher PD-L1 expression, which may be helpful for screening patients with different immune statuses to guide effective clinical treatment combinations.
RESUMO
Global obesity epidemics impacts human health and causes obesity-related illnesses, including the obesity-related kidney and liver diseases. UTX, a histone H3K27 demethylase, plays important roles in development and differentiation. Here we show that kidney-specific knockout Utx inhibits high-fat diet induced lipid accumulation in the kidney and liver via upregulating circulating serine levels. Mechanistically, UTX recruits E3 ligase RNF114 to ubiquitinate phosphoglycerate dehydrogenase, the rate limiting enzyme for de novo serine synthesis, at Lys310 and Lys330, which leads to its degradation, and thus suppresses renal and circulating serine levels. Consistently, phosphoglycerate dehydrogenase and serine levels are markedly downregulated in human subjects with diabetic kidney disease or obesity-related renal dysfunction. Notably, oral administration of serine ameliorates high-fat diet induced fatty liver and renal dysfunction, suggesting a potential approach against obesity related metabolic disorders. Together, our results reveal a metabolic homeostasis regulation mediated by a renal UTX-PHGDH-serine axis.
Assuntos
Nefropatias Diabéticas , Doenças Metabólicas , Histona Desmetilases , Humanos , Rim , Fígado , Obesidade/complicações , Fosfoglicerato Desidrogenase/genética , SerinaRESUMO
Rationale: Ischemia-reperfusion (IR) induced acute kidney injury (AKI) causes serious clinical problems associated with high morbidity and mortality. Mecp2 is a methyl-CpG binding protein, its mutation or deletion causes a neurodevelopment disease called Rett syndrome. Notably, some Rett syndrome patients present urological dysfunctions. It remains unclear whether and how Mecp2 affects AKI. Methods: Renal tubular cell specific Mecp2 deletion mice challenged with IR injury were used to investigate the effects of Mecp2 on renal tubular damage, function, cell death, fibrosis and inflammation. Cultured renal epithelial cell lines were transfected with wildtype or different domain-deletion mutants of Mecp2 to study the effects of Mecp2 on Il-6/STAT3 signaling. Results: Our results indicated rapidly upregulated Mecp2 upon acute in vivo and in vitro renal injury. Notably, increased tubular MeCP2 staining was also found in the renal sections of AKI patients. Furthermore, ablation of Mecp2 aggravated renal injury, and promoted renal cell death, inflammation, and fibrosis. Mechanistically, through its transcriptional repression domain, Mecp2 bound to the promoter of proinflammatory cytokine Il-6 to negatively regulate its expression, thus inhibiting STAT3 activation. Conclusions: A novel protective role of Mecp2 against AKI via repressing the Il-6/STAT3 axis was suggested.
Assuntos
Injúria Renal Aguda , Traumatismo por Reperfusão , Síndrome de Rett , Injúria Renal Aguda/genética , Injúria Renal Aguda/metabolismo , Animais , Apoptose , Fibrose , Humanos , Inflamação/metabolismo , Interleucina-6/metabolismo , Rim/patologia , Proteína 2 de Ligação a Metil-CpG/genética , Proteína 2 de Ligação a Metil-CpG/metabolismo , Proteína 2 de Ligação a Metil-CpG/farmacologia , Camundongos , Camundongos Endogâmicos C57BL , Traumatismo por Reperfusão/metabolismo , Síndrome de Rett/metabolismo , Fator de Transcrição STAT3/metabolismoRESUMO
Background: Serous borderline ovarian tumour (SBOT) is the most common type of BOT. Fertility sparing surgery (FSS) is an option for patients with SBOT, though it may increase the risk of recurrence. The clinical and molecular features of its recurrence are important and need to be investigated in detail. Methods: An internal cohort of 319 patients with SBOT was collected from Aug 1, 2009 to July 31, 2019 from the Obstetrics and Gynecology Hospital of Fudan University in China. An external cohort of 100 patients with SBOT was collected from Aug 1, 2009 to Nov 30, 2019 from the Shandong Provincial Hospital in China. The risk factors for the recurrence were identified by multivariate cox analysis. Several computational methods were tested to establish a prediction tool for recurrence. Whole genome sequencing, RNA-seq, metabolomics and lipidomics were used to understand the molecular characteristics of the recurrence of SBOT. Findings: Five factors were significantly correlated with SBOT recurrence in a Han population: micropapillary pattern, advanced stage, FSS, microinvasion, and lymph node invasion. A random forest-based online recurrence prediction tool was established and validated using an internal cohort and an independent external cohort for patients with SBOT. The multi-omics analysis on the original SBOT samples revealed that recurrence is related to metabolic regulation of immunological suppression. Interpretation: Our study identified several important clinical and molecular features of recurrent SBOT. The prediction tool we established could help physicians to estimate the prognosis of patients with SBOT. These findings will contribute to the development of personalised and targeted therapies to improve prognosis. Funding: JL was funded by MOST 2020YFA0803600, 2018YFA0801300, NSFC 32071138, and SKLGE-2118 to Jin Li; JY was funded by the Initial Project for Young and Middle-aged Medical Talents of Wuhan City, Hubei Province ([2014] 41); HH was funded by MOST 2019YFA0801900 and 2020YF1402600 to He Huang; JS was funded by NSFC 22,104,080; CG was funded by Natural Science Foundation of Shanghai 20ZR1408800 and NSFC82171633; BL was funded by Natural Science Foundation of Shanghai 19ZR1406800.
RESUMO
BACKGROUND: Liver metastases (LM) are the most common tumors encountered in the liver and continue to be a significant cause of morbidity and mortality. Identification of the primary tumor of any LM is crucial for the implementation of effective and tailored treatment approaches, which still represents a difficult problem in clinical practice. METHODS: The resection or biopsy specimens and associated clinicopathologic data were archived from seven independent centers between January 2017 and December 2020. The primary tumor sites of liver tumors were verified through evaluation of available medical records, pathological and imaging information. The performance of a 90-gene expression assay for the determination of the site of tumor origin was assessed. RESULT: A total of 130 LM covering 15 tumor types and 16 primary liver tumor specimens that met all quality control criteria were analyzed by the 90-gene expression assay. Among 130 LM cases, tumors were most frequently located in the colorectum, ovary and breast. Overall, the analysis of the 90-gene signature showed 93.1% and 100% agreement rates with the reference diagnosis in LM and primary liver tumor, respectively. For the common primary tumor types, the concordance rate was 100%, 95.7%, 100%, 93.8%, 87.5% for classifying the LM from the ovary, colorectum, breast, neuroendocrine, and pancreas, respectively. CONCLUSION: The overall accuracy of 93.8% demonstrates encouraging performance of the 90-gene expression assay in identifying the primary sites of liver tumors. Future incorporation of the 90-gene expression assay in clinical diagnosis will aid oncologists in applying precise treatments, leading to improved care and outcomes for LM patients.
RESUMO
Immune checkpoint blockade (ICB) has revolutionized cancer therapy. However, the response of patients to ICB is difficult to predict. Here, we examined 81 patients with lung cancer under ICB treatment and found that patients with MET amplification were resistant to ICB and had a poor progression-free survival. Tumors with MET amplifications had significantly decreased STING levels and antitumor T-cell infiltration. Furthermore, we performed deep single-cell RNA sequencing on more than 20,000 single immune cells and identified an immunosuppressive signature with increased subsets of XIST- and CD96-positive exhausted natural killer (NK) cells and decreased CD8+ T-cell and NK-cell populations in patients with MET amplification. Mechanistically, we found that oncogenic MET signaling induces phosphorylation of UPF1 and downregulates tumor cell STING expression via modulation of the 3'-UTR length of STING by UPF1. Decreased efficiency of ICB by MET amplification can be overcome by inhibiting MET. SIGNIFICANCE: We suggest that the combination of MET inhibitor together with ICB will overcome ICB resistance induced by MET amplification. Our report reveals much-needed information that will benefit the treatment of patients with primary MET amplification or EGFR-tyrosine kinase inhibitor resistant-related MET amplification.This article is highlighted in the In This Issue feature, p. 2659.
Assuntos
Neoplasias Pulmonares , Proteínas Proto-Oncogênicas c-met , Linfócitos T CD8-Positivos , Amplificação de Genes , Humanos , Imunoterapia , Células Matadoras Naturais , Neoplasias Pulmonares/terapia , Proteínas de Membrana/antagonistas & inibidores , Proteínas de Membrana/genética , Proteínas de Membrana/metabolismo , Oncogenes , Proteínas Proto-Oncogênicas c-met/genética , Proteínas Proto-Oncogênicas c-met/metabolismoRESUMO
Pathologists generally pan, focus, zoom and scan tissue biopsies either under microscopes or on digital images for diagnosis. With the rapid development of whole-slide digital scanners for histopathology, computer-assisted digital pathology image analysis has attracted increasing clinical attention. Thus, the working style of pathologists is also beginning to change. Computer-assisted image analysis systems have been developed to help pathologists perform basic examinations. This paper presents a novel lightweight detection framework for automatic tumor detection in whole-slide histopathology images. We develop the Double Magnification Combination (DMC) classifier, which is a modified DenseNet-40 to make patch-level predictions with only 0.3 million parameters. To improve the detection performance of multiple instances, we propose an improved adaptive sampling method with superpixel segmentation and introduce a new heuristic factor, local sampling density, as the convergence condition of iterations. In postprocessing, we use a CNN model with 4 convolutional layers to regulate the patch-level predictions based on the predictions of adjacent sampling points and use linear interpolation to generate a tumor probability heatmap. The entire framework was trained and validated using the dataset from the Camelyon16 Grand Challenge and Hubei Cancer Hospital. In our experiments, the average AUC was 0.95 in the test set for pixel-level detection.
Assuntos
Processamento de Imagem Assistida por Computador , Metástase Neoplásica/diagnóstico , Patologia , Humanos , Metástase Neoplásica/patologiaRESUMO
OBJECTIVE: A positive result of SARS-CoV-2 nucleic acid detection provides critical laboratory evidence for clinical confirmed diagnosis, pandemic status evaluation, a pandemic prevention plan, treatment of infected people with symptoms, and protection of uninfected people. This study aims to provide a practical reference for SARS-CoV-2 nucleic acid-related research and detection. METHODS: Our laboratory has established policies combining personnel management and quality control practices for SARS-CoV-2 nucleic acid detection during the pandemic. RESULTS: In this article, we describe cross-department personnel management and key points of personal protection and quality control in the testing process. We also report on the differences in detection and the compatibility between different brand kits. CONCLUSION: It is critical to maintain a standard and accurate laboratory operation for nucleic acid testing.
Assuntos
Teste de Ácido Nucleico para COVID-19/normas , COVID-19/diagnóstico , COVID-19/epidemiologia , Pandemias , RNA Viral/genética , SARS-CoV-2/genética , COVID-19/virologia , Humanos , Variações Dependentes do Observador , Equipamento de Proteção Individual/provisão & distribuição , Gestão de Recursos Humanos/métodos , Garantia da Qualidade dos Cuidados de Saúde/métodos , Controle de Qualidade , Kit de Reagentes para DiagnósticoRESUMO
Coronavirus disease 2019 (COVID-19) is an infectious disease caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). To counter COVID-19 spreading, an infrastructure to provide rapid and thorough molecular diagnostics and serology testing is the cornerstone of outbreak and pandemic management. We hereby review the clinical insights with regard to using molecular tests and immunoassays in the context of COVID-19 management life cycle: the preventive phase, the preparedness phase, the response phase and the recovery phase. The spatial and temporal distribution of viral RNA, antigens and antibodies during human infection is summarized to provide a biological foundation for accurate detection of the disease. We shared the lessons learned and the obstacles encountered during real world high-volume screening programs. Clinical needs are discussed to identify existing technology gaps in these tests. Leverage technologies, such as engineered polymerases, isothermal amplification, and direct amplification from complex matrices may improve the productivity of current infrastructure, while emerging technologies like CRISPR diagnostics, visual end point detection, and PCR free methods for nucleic acid sensing may lead to at-home tests. The lessons learned, and innovations spurred from the COVID-19 pandemic could upgrade our global public health infrastructure to better combat potential outbreaks in the future.
Assuntos
COVID-19/diagnóstico , COVID-19/imunologia , Imunoensaio/métodos , Patologia Molecular/métodos , Animais , Humanos , Estágios do Ciclo de Vida , Pandemias/prevenção & controle , SARS-CoV-2/imunologia , Testes Sorológicos/métodosRESUMO
OBJECTIVES: The study was aimed to analyze the clinicopathological and molecular pathological features of diffuse large B-cell lymphoma (DLBCL) in the oropharyngeal and maxillofacial region. SUBJECTS AND METHODS: A retrospective review was performed with 36 patients who were diagnosed with primary DLBCL of the oropharyngeal and maxillofacial region from 2009 to 2017 in the Department of Pathology at the Hospital of Stomatology, Wuhan University. Immunohistochemistry and fluorescence in situ hybridization were performed. RESULTS: Gene rearrangements of BCL2, BCL6, and MYC were observed in 5.6%, 33.3%, and 22.2%, respectively, including two double-hit and one triple-hit DLBCL (8.3%). There was a significant correlation between MYC protein expression and gene translocation (rs = 0.679, p < .001). However, 25% of cases with MYC rearrangement showed low MYC protein expression. In univariate analysis, MYC protein expression, BCL2 rearrangement, MYC rearrangement, and double/triple-hit DLBCL were associated with shorter overall survival, whereas only MYC protein expression was an independent prognostic value in multivariate model. CONCLUSIONS: MYC protein expression was an essential prognostic marker of DLBCL in the oropharyngeal and maxillofacial region. Notably, immunohistochemical staining of MYC, BCL2, and BCL6 could not predict their gene rearrangements, although MYC protein expression was correlated with gene translocation.