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Analysis of exosomes provides important information for rapid and non-invasive screening of tumors. However, sensitive and convenient detection of exosomes remains technically challenging to date. Herein, a colorimetric aptasensor based on the light-stimulated oxidase-mimicking activity of FITC was constructed for detecting ovarian cancer (OC) exosomes. The aptasensor contained an EpCAM aptamer to capture OC exosomes. Cholesterol and fluorescein (FITC) were used to modify either end of the DNA (DNA anchor). The DNA anchor could combine with exosomes through a hydrophobic reaction between cholesterol and the lipid membrane. FITC oxidized 3,3',5,5'-tetramethylbenzidine (TMB) under a 365 nm LED light source in a temporally controllable manner under mild conditions, causing the solution to change from colorless to blue, and the corresponding UV-vis absorbance increased. Based on this principle, the exosomes were qualitatively analyzed by observing the color change with the naked eye. In parallel, the exosome concentration was also detected using UV-vis spectrophotometry. The linear range was from 2 × 105 to 100 × 105 particles per mL with a limit of detection of 1.77 × 105 particles per mL. The developed aptasensor also exhibited favorable selectivity and could discriminate the exosomes from OC cells and normal cells. Besides, the receiver operating characteristic (ROC) curve demonstrates that it is possible to distinguish between patients with OC and healthy donors (HDs) using exosomes as the biomarker. Our technology may expand the applications of DNA-based detection method-enabled OC diagnostic tools.
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Aptámeros de Nucleótidos , Técnicas Biosensibles , Colorimetría , Exosomas , Exosomas/química , Exosomas/metabolismo , Humanos , Colorimetría/métodos , Aptámeros de Nucleótidos/química , Técnicas Biosensibles/métodos , Femenino , Neoplasias Ováricas , Oxidorreductasas/química , Oxidorreductasas/metabolismo , Luz , Límite de Detección , Fluoresceína/química , Bencidinas/química , Línea Celular TumoralRESUMEN
Across cervical squamous and glandular lesions, a spectrum of human papillomavirus (HPV) genotypes has been identified. This review aims to provide a comprehensive summary detailing the distribution and profile of HPV genotypes detected in cervical lesions, leveraging insights from histological and cytological findings. High-risk HPV (HR-HPV) genotypes exhibit varying degrees of oncogenic potential, with HPV16 and HPV18 identified as the most prevalent and oncogenic types. The distribution of HR-HPV genotypes varies among different degrees of the cervical lesions and varies between squamous and glandular neoplasia. HPV16 is predominantly associated with severe lesions (precancers and carcinomas), while HPV18 demonstrates a significantly higher prevalence in endocervical as compared with squamous neoplasia. The distribution of HR-HPV in severe squamous lesions is complex, involving many HR-HPV genotypes in addition to HPV16, while the distribution of HR-HPV genotypes in endocervical glandular lesions is mainly limited in HPV18 and HPV16. Large datasets from China have identified the three most common HR-HPV genotypes in this population as stratified by diagnostic category: HPV52, HPV16, HPV58 in histologically negative cases and cervical intraepithelial neoplasia 1 (CIN1); HPV16, HPV52, HPV58 in CIN2/3; HPV16, HPV58, HPV52 or HPV18 in squamous cell carcinoma (SCC); HPV16, HPV18 and HPV52 in endocervical adenocarcinoma in situ (AIS), invasive adenocarcinoma, as well as mixed squamous and glandular lesions. HPV33 is the fourth most common HPV type in CIN2/3 and SCC, while HPV45 occurs more commonly in AIS and adenocarcinoma, compared with squamous lesions. The prevalence and distribution of multiple HR-HPV coinfections vary across different cervical diseases. The clinical significance and pathogenesis of these multiple HR-HPV infections remain uncertain, although recent two large studies demonstrate that multiple HR-HPV infections are not associated with cumulatively higher risk of high-grade cervical squamous lesion development, suggesting competitive and/or cooperative interactions among HPV genotypes. Extensive HPV genotyping aids in risk assessment and optimising clinical approaches for women with mild abnormalities in Pap cytology. Women with atypical squamous cells of undetermined significance (ASC-US) and low-grade squamous intraepithelial lesion (LSIL) Pap test results and with the infection of some HR-HPV genotypes carry a very low risk of high-grade cervical lesions. HPV genotyping can allow for risk stratification and triage optimisation for these HR-HPV-positive women. Women with atypical glandular cell (AGC) Pap test results showed a specific HPV genotyping pattern and extended HPV genotyping may be helpful for the clinical management of AGCs. Continual advancements in clinical guidelines integrating extended genotyping would increase diagnostic accuracy and refine strategies in clinical management.
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TRPS1 is aberrantly expressed in a variety of tumors, including breast, prostate, and gastric cancers, and is strongly associated with tumorigenesis or prognosis. However, the role of TRPS1 in high grade serous ovarian carcinoma (HGSC) is unknown. We investigated the relationship between TRPS1 expression and clinicopathology in HGSC patients. The tumor-related regulatory mechanisms of TRPS1 was explored through in vivo and vitro experiments. The results showed that TRPS1 was highly expressed in HGSC compared to normal tissues. It was also linked to the cell proliferation index Ki67 and poor prognosis. In vivo experiments showed that knockdown of TRPS1 could inhibit tumor growth. In vitro experiments, knockdown of TRPS1 inhibited the proliferation of ovarian cancer cells. TRPS1 exerted its regulatory role as a transcription factor, binding to the PSAT1 promoter and promoting the expression of PSAT1 gene. Meanwhile, PSAT1 was positively correlated with CCND1 expression. These results suggest that TRPS1 affects HGSC proliferation and cell cycle by regulating PSAT1 and thus CCND1 expression.
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Cistadenocarcinoma Seroso , Neoplasias Ováricas , Masculino , Femenino , Humanos , Cistadenocarcinoma Seroso/patología , Neoplasias Ováricas/patología , Factores de Transcripción/genética , Pronóstico , Proliferación Celular , Proteínas Represoras/genéticaRESUMEN
BACKGROUND: Steroid-resistant nephrotic syndrome (SRNS) is characterized by unrelieved proteinuria after an initial 4-8 weeks of glucocorticoid therapy. Genes in podocytes play an important role in causing SRNS. OBJECTIVE: This study aimed to report a pathogenic mutation in SRNS patients and investigate its effects on podocytes, as well as the pathogenic mechanism. METHODS: We screened out a novel mutation by using whole-exon sequencing in the SRNS cohort and verified it via Sanger sequencing. Conservative analysis and bioinformatic analysis were used to predict the pathogenicity of the mutation. In vitro, stable podocyte cell lines were constructed to detect the effect of the mutation on the function of the podocyte. Moreover, an in vivo mouse model of podocyte ANLN gene knockout (ANLNpodKO) was used to confirm clinical manifestations. Transcriptome analysis was performed to identify differential gene expression and related signaling pathways. RESULTS: ANLN E841K was screened from three unrelated families. ANLN E841K occurred in the functional domain and was predicted to be harmful. The pathological type of A-II-1 renal biopsy was minimal change disease, and the expression of ANLN was decreased. Cells in the mutation group showed disordered cytoskeleton, faster cell migration, decreased adhesion, increased endocytosis, slower proliferation, increased apoptosis, and weakened interaction with CD2 association protein. ANLNpodKO mice exhibited more obvious proteinuria, more severe mesangial proliferation, glomerular atrophy, foot process fusion, and increased tissue apoptosis levels than ANLNflox/flox mice after tail vein injection of adriamycin. Upregulated differentially expressed genes in cells of the mutation group were mainly enriched in the PI3K-AKT pathway. CONCLUSION: The novel mutation known as ANLN E841K affected the function of the ANLN protein by activating the PI3K/AKT/mTOR/apoptosis pathway, thus resulting in structural and functional changes in podocytes. Our study indicated that ANLN played a vital role in maintaining the normal function of podocytes. Video Abstract.
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Proteínas de Microfilamentos , Síndrome Nefrótico , Podocitos , Animales , Humanos , Ratones , Mutación/genética , Síndrome Nefrótico/genética , Síndrome Nefrótico/tratamiento farmacológico , Síndrome Nefrótico/patología , Fosfatidilinositol 3-Quinasas/metabolismo , Podocitos/patología , Proteinuria , Proteínas Proto-Oncogénicas c-akt/metabolismo , Proteínas de Microfilamentos/genéticaRESUMEN
BACKGROUND: Myeloid-derived suppressor cells (MDSCs) are capable of effectively repressing immune responses against tumors and orchestrating the tumor microenvironment, which can promote tumor angiogenesis and metastasis. The pathway networks used to modulate tumor-expanded MDSC accumulation and function remain unclear. This study identified microRNA-211 (miR-211), whose expression was significantly decreased by factors derived from tumors. METHODS: miR-211 was assumed to be critical in modulating the accumulation and activity of MDSCs isolated from ovarian cancer (OC)-bearing mice by targeting C/EBP homologous protein (CHOP). RESULTS: The upregulation of miR-211 repressed MDSC proliferation, inhibited MDSC immunosuppressive functions, and increased the number of co-incubated CD4+ and CD8+ cells. Furthermore, overexpression of miR-211 led to decreased activities of the NF-κB, PI3K/Akt, and STAT3 pathways and the subsequent downregulation of matrix metalloproteinases to promote tumor cell invasion and metastasis. CHOP overexpression counteracted the effects of miR-211 elevation on these phenotypic changes. Upregulation of miR-211 also dramatically impaired the activity of MDSCs and suppressed OC tumor growth in vivo. CONCLUSION: These results indicated that the miR-211-CHOP axis in MDSCs plays an essential role in the metastasis and proliferation of tumor-expanded MDSCs and might represent a promising cancer treatment target.
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MicroARNs , Células Supresoras de Origen Mieloide , Neoplasias , Animales , Ratones , MicroARNs/genética , MicroARNs/metabolismo , Células Mieloides , Fosfatidilinositol 3-Quinasas/metabolismo , Neoplasias/patología , Proliferación Celular , Microambiente Tumoral/genéticaRESUMEN
Objective: To investigate the clinicopathological characteristics, molecular genetic characteristics and prognosis of extraventricular neurocytoma located in the sellar/suprasellar region. Methods: Seven archived tumor samples derived from 4 patients with neurocytoma in the sellar/suprasellar region were collected from the First Affiliated Hospital of Fujian Medical University and the Affiliated Hospital of Qingdao University and retrospectively analyzed for clinical manifestations, imaging features, and histopathological features. Neuronal and pituitary biomarkers and molecular features were detected in these tumor tissues by immunohistochemistry and FISH or Sanger sequencing. The related literature was reviewed. Results: Three patients were female, while 1 was male, with an average age of 35.5 years (range: 27 to 45 years). The initial manifestations were mainly headache and blurred vision in both eyes. The first MRI examination showed marginally enhancing masses in the intrasellar or intra- to suprasellar region. The diagnosis of pituitary adenomas was based on imaging features. The levels of pituitary hormones were normal. Histologically, the tumor cells were arranged in a sheet-like, monotonous architecture and were uniform in size and shape with round to oval, exquisite and hyperchromatic nuclei, which densely packed close to one another and were separated only by a delicate neuropil background. There was no evident mitosis, necrosis or microvascular proliferation. The three cases of recurrent tumors were highly cellular and showed increased mitotic activity. Immunohistochemically, the tumor cells were positive for syn, CR, CgA, and vasopressin and were focally positive for NeuN, TTF-1, NF, CK8, vimentin, and S100 proteins. Other markers, including IDH1, BRAF VE1, Olig-2, and EMA, were negative. Pituitary transcription factors and anterior pituitary hormones were negative. Molecular genetic testing showed that the tumor cells lacked IDH gene mutations, LOH of 1p/19q, MYCN amplification, and EGFR alteration. With a median follow-up of 74.5 months (range 23 to 137 months), 3 patients relapsed at 11, 50, and 118 months after the initial surgery. Conclusion: The morphological features and immunophenotypes of neurocytoma in the sellar/suprasellar region are similar to those of classic central neurocytoma. The prognosis is relatively good. Gross-subtotal resection and atypical subtype may be related to tumor recurrence.
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Adenoma , Neoplasias Encefálicas , Neurocitoma , Neoplasias Hipofisarias , Neoplasias de los Tejidos Blandos , Adenoma/patología , Neoplasias Encefálicas/diagnóstico por imagen , Neoplasias Encefálicas/genética , Femenino , Humanos , Masculino , Recurrencia Local de Neoplasia , Neurocitoma/diagnóstico , Neurocitoma/genética , Neurocitoma/metabolismo , Neoplasias Hipofisarias/diagnóstico , Neoplasias Hipofisarias/genética , Neoplasias Hipofisarias/metabolismo , Estudios RetrospectivosRESUMEN
Long noncoding RNAs (lncRNAs) have been found to play an important role in the occurrence and development of endometrial carcinoma (EC). Here, using RNA sequencing analysis, we systemically screened and identified the lncRNA eukaryotic translation initiation factor 1A, X-linked (EIF1AX)-AS1, which is aberrantly downregulated in clinical EC tissues and closely correlated with tumor type. EIF1AX-AS1 markedly inhibited EC cell proliferation and promoted apoptosis in vitro and in vivo. Mechanistically, EIF1AX-AS1 interacts with EIF1AX mRNA and poly C binding protein 1 (PCBP1), which promote EIF1AX mRNA degradation. Intriguingly, by interacting with internal ribosome entry site-related protein Y-box binding protein 1 (YBX-1), EIF1AX promotes c-Myc translation through the internal ribosome entry site pathway. c-Myc promotes EIF1AX transcription and thus forms a feed-forward loop to regulate EC cell proliferation. Taken together, these data reveal new insights into the biology driving EC proliferation and highlights the potential of lncRNAs as biomarkers for prognosis and future therapeutic targets for cancer.
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Neoplasias Endometriales , ARN Largo no Codificante , Apoptosis/genética , Línea Celular Tumoral , Proliferación Celular/genética , Neoplasias Endometriales/metabolismo , Femenino , Regulación Neoplásica de la Expresión Génica , Humanos , Sitios Internos de Entrada al Ribosoma , ARN Largo no Codificante/genética , ARN Largo no Codificante/metabolismo , ARN Mensajero/genética , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
Dysregulation of eukaryotic translation initiation factor 1A, X-linked (EIF1AX), has been implicated in the pathogenesis of some cancers. However, the role of EIF1AX in endometrial carcinoma (EC) remains unknown. We investigated the EIF1AX expression in EC patients and assessed its tumorigenesis-associated function and nucleocytoplasmic transport mechanism in vitro and in vivo. The results indicated that the cytoplasmic EIF1AX expression showed a gradual increase when going from endometrium normal tissue, simple endometrial hyperplasia, complex endometrial hyperplasia, and endometrial atypical hyperplasia to EC, while vice versa for the nuclear EIF1AX expression. In addition, the cytoplasmic EIF1AX expression was positively correlated with histologic type, high International Federation of Gynecology and Obstetrics (FIGO) grade, advanced FIGO stage, deeper infiltration, high Ki67 index, and shorter recurrence-free survival in EC patients. In vitro, short hairpin RNA-mediated EIF1AX depletion or SV40NLS-mediated EIF1AX import into the nucleus in multiple human EC cells potently suppressed cell migration and invasion, epithelial-mesenchymal transition, and lung metastasis. Moreover, exportin 1 induced the transport of EIF1AX from the nucleus to the cytoplasm that could be inhibited by leptomycin B treatment or the mutation in the EIF1AX location sequence. These results demonstrate that cytoplasmic EIF1AX may play a key role in the incidence and promotion of EC, and thus, targeting EIF1AX or its nucleocytoplasmic transport process may offer an effective new therapeutic approach to EC.
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Hiperplasia Endometrial , Neoplasias Endometriales , Factor 1 Eucariótico de Iniciación , Receptores Citoplasmáticos y Nucleares , Femenino , Humanos , Línea Celular Tumoral , Proliferación Celular , Citoplasma/metabolismo , Hiperplasia Endometrial/metabolismo , Hiperplasia Endometrial/patología , Neoplasias Endometriales/genética , Endometrio/metabolismo , Receptores Citoplasmáticos y Nucleares/metabolismo , Factor 1 Eucariótico de Iniciación/metabolismo , Proteína Exportina 1RESUMEN
Protein phosphatase 2A (PP2A) is one of the major protein serine/threonine phosphatases (PPPs) with regulatory effects on several cellular processes, but its role and function in Adriamycin (ADR)-treated podocytes injury needs to be further explored. Mice podocytes were treated with ADR and PP2A inhibitor (okadaic acid, OA). After transfection, cell apoptosis was detected by flow cytometry. Expressions of podocytes injury-, apoptosis- and epithelial-to-mesenchymal transition (EMT)- and JNK-interacting protein 4/p38-Mitogen-Activated Protein Kinase (JIP4/p38-MAPK) pathway-related factors were measured using quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot as needed. Interaction between PP2A and JIP4/MAPK pathway was confirmed using co-immunoprecipitation (Co-Ip) assay. In podocytes, ADR inhibited PP2A, Nephrin and Wilms' tumor (WT) 1 expressions yet upregulated apoptosis and Desmin expression, and suppressing PP2A expressionenhanced the effects. PP2A overexpression reversed the effects of ADR on PP2A and podocyte injury-related factors expressions and apoptosis of podocytes. JIP4 was the candidate gene interacting with both PP2A and p38-MAPK pathway, and PP2A overexpression alleviated the effects of ADR on p38-MAPK pathway-related factors expressions. Additionally, in ADR-treated podocytes, PP2A suppression enhanced the effects of ADR, yet silencing of JIP4 reversed the effects of PP2A suppression on regulating p38-MAPK pathway-, apoptosis- and EMT-related factors expressions and apoptosis, with upregulations of B-cell lymphoma-2 (Bcl-2) and E-cadherin and down-regulations of Bcl-2 associated protein X (Bax), cleaved (C)-casapse-3, N-cadherin, Vimentin and Snail. PP2A protects ADR-treated podocytes against injury and EMT by suppressing JIP4/p38-MAPK pathway, showing their interaction in podocytes.
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We studied a rabbit model of rheumatoid arthritis (RA) to examine the time course of changes in synovial neovascularization based on quantitative power Doppler ultrasound and contrast-enhanced ultrasound (CEUS). Twenty-five male New Zealand rabbits were in the ovalbumin-induced arthritis (OIA) group, and 5 were in the control group. Both rear knee joints of all rabbits were examined using conventional US and CEUS over 16 weeks. The knee synoviums of OIA rabbits were sampled by US-guided biopsy, and expression of CD31 and vascular endothelial growth factor (VEGF) was determined by immunohistochemistry. The correlation of joint damage based on multimodal US with microvessel density (CD31 positivity) and VEGF expression at different times was analyzed. OIA rabbits had increased synovial expression of CD31 and VEGF from weeks 6 to 12 (p < 0.01). During the early stage of CEUS enhancement, "dot enhancement" was more common at weeks 6 and 8, and "stripe enhancement" was more common at weeks 12 and 16 (p < 0.05). There were significant positive correlations of synovial CD31 and VEGF expression with power Doppler image grade, CEUS grade and peak intensity (p < 0.05 for all). Thus, OIA rabbits mimicked early-stage RA at 6 to 8 weeks, middle-stage RA at 8 to 12 weeks and late-stage RA at 12 to 16 weeks. Power Doppler image grade, CEUS grade and peak intensity, especially when combined with CD31 expression data, accurately characterized the extent of synovial vascularization in a rabbit model of RA. Increased vascularity based on CEUS may have value for the early diagnosis of RA.
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Artritis Reumatoide/diagnóstico por imagen , Medios de Contraste , Aumento de la Imagen , Articulación de la Rodilla/diagnóstico por imagen , Neovascularización Patológica/diagnóstico por imagen , Membrana Sinovial/irrigación sanguínea , Membrana Sinovial/diagnóstico por imagen , Ultrasonografía Doppler/métodos , Animales , Modelos Animales de Enfermedad , Masculino , Conejos , Factores de TiempoRESUMEN
OBJECTIVE: To explore the clinical and pathologic features of ovarian juvenile granulosa cell tumors (JGCTs). METHODS: Clinical data, histopathologic observations, immunohistochemical results, FOXL2 mutation status, and follow-up information of 7 JGCT cases were studied. RESULTS: The patients most commonly presented with abdominal distension and pain (5 cases), followed by precocious puberty (1 case) and a pelvic mass (1 case). Six patients had stage I disease, and 1 had stage IV disease. The microscopic examinations typically showed lobular growth punctuated by variably sized and shaped follicles. Rare features included a reticular-cystic appearance mimicking a yolk sac tumor (2 cases), a lobular appearance similar to a sclerosing stromal tumor (1 case), strands and cords (1 case), pseudopapillary appearance (2 cases), spindle cell appearance (1 case), microcystic appearance (1 case), hobnail cells (1 case), and rhabdomyoid cells (1 case). No FOXL2 mutation was encountered. After a median follow-up of 53 months, only 1 patient with a strongly diffuse TP53-positive tumor died of the disease, and 2 successfully had babies. CONCLUSIONS: JGCT is a rare neoplasm with a wide morphologic spectrum and is easily confused with other tumors. Familiarity with the characteristics, rare atypical appearances, and immunohistochemical results may aid in obtaining a correct diagnosis.
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Tumor de Células de la Granulosa/patología , Neoplasias Ováricas/patología , Ovario/patología , Adulto , Niño , Femenino , Factores de Transcripción Forkhead/genética , Tumor de Células de la Granulosa/genética , Humanos , Recién Nacido , Mutación , Neoplasias Ováricas/genéticaRESUMEN
BACKGROUND: Female Dent disease 1 patients with low-molecular-weight proteinuria (LMWP) due to CLCN5 gene mutation were rarely reported, and these cases that the people were also with Turner syndrome (TS) were even hardly documented before. CASE PRESENTATION: Here we report a 3-year and 11-month old Chinese girl with short stature who had a karyotype of 46,X,i(X)(q10) and a de novo pathogenic variant in the CLCN5 gene on the short arm of X chromosome. Laboratory examinations showed that the patient had LMWP, hypercalciuria, hypophosphatemia, delayed bone age, and genital dysplasia. CONCLUSION: The combination of i(X)(q10) and CLCN5 mutation causes the deletion of the wild-type CLCN5 allele that results in Dent-1 and TS. To the best of our knowledge, this is the first case that a female CLCN5 mutation hemizygote is diagnosed with Dent-1 and Turner syndrome due to isochromosome X. Also, our case has indicated that the prevalence of the situation may be largely underestimated because of the mild signs of females with Dent-1.
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Canales de Cloruro/genética , Enfermedades Genéticas Ligadas al Cromosoma X/genética , Nefrolitiasis/genética , Síndrome de Turner/genética , Desarrollo Óseo , Huesos/diagnóstico por imagen , Preescolar , Femenino , Enfermedades Genéticas Ligadas al Cromosoma X/complicaciones , Enfermedades Genéticas Ligadas al Cromosoma X/fisiopatología , Hemicigoto , Humanos , Hipercalciuria/fisiopatología , Hipofosfatemia/fisiopatología , Isocromosomas , Riñón/diagnóstico por imagen , Mutación , Nefrolitiasis/complicaciones , Nefrolitiasis/fisiopatología , Ovario/anomalías , Ovario/diagnóstico por imagen , Proteinuria/fisiopatología , Síndrome de Turner/complicaciones , Síndrome de Turner/fisiopatología , Útero/anomalías , Útero/diagnóstico por imagenAsunto(s)
Artritis Reumatoide/diagnóstico por imagen , Medios de Contraste/química , Membrana Sinovial/irrigación sanguínea , Membrana Sinovial/diagnóstico por imagen , Ultrasonografía , Animales , Artritis Reumatoide/patología , Biopsia , Articulación de la Rodilla/diagnóstico por imagen , Articulación de la Rodilla/patología , Masculino , Microvasos/diagnóstico por imagen , Microvasos/patología , Conejos , Membrana Sinovial/patología , Sinovitis/diagnóstico por imagen , Sinovitis/patologíaRESUMEN
BACKGROUND: Protein phosphorylation & dephosphorylation are ubiquitous cellular processes that allow for the nuanced and reversible regulation of protein activity. Protein phosphatase 2A (PP2A) is a multifunction phosphatase that is well expressed in all cell types of kidney during early renal development, though its functions in kidney remains to be elucidated. METHODS: PP2A conditional knock-out mice was generated with PP2A fl/fl mice that were crossed with Podocin-Cre mice. The phenotype of Pod-PP2A-KO mice (homozygous for the floxed PP2A allele with Podocin-Cre) and littermate PP2A fl/fl controls (homozygous for the PP2A allele but lacking Podocin-Cre) were further studied. Primary podocytes isolated from the Pod-PP2A-KO mice were cultured and they were then employed with sing label-free nano-LC - MS/MS technology on a Q-exactive followed by SIEVE processing to identify possible target molecular entities for the dephosphorylation effect of PP2A, in which Western blot and immunofluorescent staining were used to analyze further. RESULTS: Pod-PP2A-KO mice were developed with weight loss, growth retardation, proteinuria, glomerulopathy and foot process effacement, together with reduced expression of some slit diaphragm molecules and cytoskeleton rearrangement of podocytes. Y box protein 1 (YB-1) was identified to be the target molecule for dephosphorylation effect of PP2A. Furthermore, YB-1 phosphorylation was up-regulated in the Pod-PP2A-KO mice in contrast to the wild type controls, while total and un-phosphorylated YB-1 both was moderately down-regulated in podocytes from the Pod-PP2A-KO mice. CONCLUSION: Our study revealed the important role of PP2A in regulating the development of foot processes and fully differentiated podocytes whereas fine-tuning of YB-1 via a post-translational modification by PP2A regulating its activity might be crucial for the functional integrity of podocytes and glomerular filtration barrier.
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Glomérulos Renales/citología , Glomérulos Renales/fisiología , Podocitos/citología , Proteína Fosfatasa 2/metabolismo , Animales , Peso Corporal , Citoesqueleto/metabolismo , Femenino , Regulación Enzimológica de la Expresión Génica , Técnicas de Inactivación de Genes , Humanos , Glomérulos Renales/patología , Glomérulos Renales/fisiopatología , Masculino , Ratones , Fosforilación , Podocitos/patología , Proteína Fosfatasa 2/deficiencia , Proteína Fosfatasa 2/genética , Proteinuria/enzimología , Proteinuria/genética , Proteína 1 de Unión a la Caja Y/metabolismoRESUMEN
OBJECTIVE: To evaluate the expression and prognostic significance of ALDH1A1 in gastric neuroendocrine carcinoma. MATERIALS AND METHODS: Immunohistochemical stains of ALDH1A1 were evaluated in 67 cases of gastric neuroendocrine carcinoma. The findings were correlated with clinicopathologic variables and overall survival. RESULTS: Immunohistochemistry revealed positive cytoplasmic immunoreactivity in 35 of 67 (52.2%) tumors and strongly positive immunoreactivity in 14 of 67 (20.9%). Strongly positive ALDH1A1 expression, but not positive staining, was significantly associated with lymph node status, lymphovascular invasion, and ki-67 index (P=0.039, 0.045, and 0.045, respectively). Kaplan-Meier survival curves and log-rank tests showed significantly poorer prognoses in cases of high ALDH1A1 expression compared to cases of low ALDH1A1 expression or the negative control group (MST, 17 vs. 52 months; P=0.026). Multivariate analysis showed that high ALDH1A1 expression, lymph node metastasis, and lymphovascular invasion had significant associations with decreased overall survival (P=0.029, 0.008, and 0.005, respectively). CONCLUSIONS: High ALDH1A1 expression may be a prognostic indicator of survival in patients with gastric neuroendocrine carcinoma.
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Aldehído Deshidrogenasa/metabolismo , Carcinoma Neuroendocrino/metabolismo , Regulación Neoplásica de la Expresión Génica/genética , Neoplasias Gástricas/metabolismo , Adulto , Anciano , Familia de Aldehído Deshidrogenasa 1 , Biomarcadores de Tumor/biosíntesis , Carcinoma Neuroendocrino/mortalidad , Carcinoma Neuroendocrino/patología , Línea Celular Tumoral , Femenino , Humanos , Masculino , Persona de Mediana Edad , Pronóstico , Retinal-Deshidrogenasa , Neoplasias Gástricas/mortalidad , Neoplasias Gástricas/patologíaRESUMEN
Distal renal tubular acidosis (dRTA) is characterized by metabolic acidosis due to uric acid dysfunction. The aim of this study was to demonstrate the genetic diagnosis of Chinese children with dRTA by whole-exome sequencing. From Jan. 2010 to Sept. 2015, 16 children with dRTA were recruited to investigate the possibility of genetic diagnosis and to examine any genotype-phenotype relationships in these patients. Sanger sequencing was used to confirm mutations identified by whole-exome sequencing. Clinical and biological features in the patients included hyperchloremic metabolic acidosis, impaired growth, hypokalemia, nephrocalcinosis, nephrolithiasis, hypercalciuria, hypocitraturia, and rickets or osteomalacia. Seventeen mutations in the solute carrier family 4 member 1 (SLC4A1), ATPase H+ transporting V0 subunit a4 (ATP6V0A4), ATPase H+ transporting V1 subunit B1 (ATP6V1B1), WNK lysine deficient protein kinase 1 (WNK1) and the claudin 16 (CLDN16) were identified in 15 patients, and 14 of these mutations are novel. Only 1 patient was negative for any mutations. Our results demonstrate the existence of SLC4A1, ATP6V1B1, ATP6V0A4, WNK1 and CLDN16 mutations in Chinese children with dRTA and indicate that compound heterozygosity at 2 or more different but related genes can be responsible for its pathogenesis. This study also indicates that whole-exome sequencing is a labor and cost-effective means of analyzing dRTA-associated genes.
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AIM OF THE STUDY: To determine the significance of expression of synaptophysin, chromogranin A, and Ki-67 and their association with clinicopathological parameters, and to find out the possible prognostic factors in gastric neuroendocrine carcinoma (G-NEC). MATERIAL AND METHODS: We investigated the immunohistochemical features and prognosis of 62 G-NECs, and evaluated the association among expressions of synaptophysin, chromogranin A, and Ki-67, clinicopathological variables, and outcome. RESULTS: Chromogranin A expression was found more commonly in small-cell NECs (9/9, 100%) than in large-cell NECs (27/53, 51%) (p = 0.008). No statistical significance was found in Ki-67 (p = 0.494) or synaptophysin (p > 0.1) expression between NEC cell types. Correlation analyses revealed that Ki-67 expression was significantly associated with mid-third disease of stomach (p = 0.005) and vascular involvement (p = 0.006), and had a trend of significant correlation with tumour relapse (p = 0.078). High expression of chromogranin A was significantly associated with histology of small-cell NECs (p = 0.008) and lesser tumour greatest dimension (p = 0.038). The prognostic significance was determined by means of Kaplan-Meier survival estimates and log-rank tests, and as a result, early TNM staging and postoperative chemotherapy were found to be correlated with longer overall survival (p < 0.05). Univariate analysis revealed associations between poor prognosis in NECs and several factors, including high TNM staging (p = 0.048), vascular involvement (p = 0.023), relapse (p = 0.004), and microscopic/macroscopic residual tumour (R1/2, p < 0.001). In a multivariate analysis, relapse was identified as the sole independent prognostic factor. CONCLUSIONS: No significant correlation between survival and expression of synaptophysin, chromogranin A, or Ki-67 has been determined in G-NECs. Our study indicated that early diagnosis, no-residual-tumour resection, and postoperative chemotherapy were possible prognostic factors.
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OBJECTIVE: To study the clinicopathologic features of delayed radiation-induced brain injury after radiotherapy for brain tumor. METHODS: The clinical, histopathologic and immunohistochemical features of 9 cases with delayed radiation-induced injury were evaluated. RESULTS: The disease occurred from 6 months to 12 years after radiotherapy and often presented with headache and muscle weakness. Magnetic resonance imaging showed peripheral enhancing lesions with slight mass effect and surrounding edema. Microscopically, the major changes included coagulative necrosis, fibrinoid necrosis of vessels, vascular hyalinization with luminal stenosis and peripheral reactive gliosis. Immunostaining for hypoxia-inducible factors 1α was positive in reactive astrocytes. CONCLUSIONS: Delayed radiation-induced brain injury is a relatively common complication of radiation therapy. The lesion was frequently misdiagnosed as brain tumor. Correct diagnosis relies on clinical, radiologic and pathologic correlation.
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Neoplasias Encefálicas/radioterapia , Glioma/radioterapia , Traumatismos por Radiación/patología , Radioterapia/efectos adversos , Anciano , Femenino , Humanos , Subunidad alfa del Factor 1 Inducible por Hipoxia/metabolismo , Imagen por Resonancia Magnética , Masculino , Persona de Mediana Edad , Traumatismos por Radiación/diagnóstico , Traumatismos por Radiación/diagnóstico por imagen , Traumatismos por Radiación/metabolismo , Traumatismos por Radiación/cirugía , Tomografía Computarizada por Rayos XRESUMEN
Ependymoma with cartilaginous metaplasia with or without bone formation is exceedingly rare. Only eight cases have been reported in the literature. We report a case of ependymoma with cartilaginous and osseous metaplasia in a 5-year-old boy. Microscopically, the tumor was composed of neoplastic ependymal tissue and mature cartilage and bone. Immunohistochemically, glial fibrillary acidic protein and epithelial membrane antigen were positive for ependymoma cells but negative for cartilage and bone. Recurrence occurred after 15-month follow-up. The patient deteriorated rapidly and died after 1 month. Reviewing 8 reported cases and our latest case, we found that 3 cases of ependymoma with cartilaginous metaplasia were treated with radiotherapy. Six cases had recurrence from 6 months to 8 years and 2 cases died on the day of operation. These findings suggest that ependymoma with cartilaginous metaplasia might have more aggressive clinical behavior.
Asunto(s)
Neoplasias Encefálicas/patología , Cartílago/patología , Ependimoma/patología , Neoplasias Encefálicas/cirugía , Preescolar , Ependimoma/cirugía , Resultado Fatal , Cuarto Ventrículo/patología , Humanos , Masculino , MetaplasiaRESUMEN
OBJECTIVE: To investigate the specific genetic alterations in nasal NK/T cell lymphoma (N-NK/T-L) and the significance thereof. METHODS: Restriction landmark genomic scanning (RLGS) was performed on the specimen of a case of N-NK/T-L and the peripheral blood leukocytes of the same case. The RLGS image was analyzed with the Virtual genome scan (VGS) software so as to discover abnormality of T-bet gene. The discovered abnormal gene underwent cloning and preparation of probes. Southern blotting and dot blotting were performed on 3 cases of fresh N-NK/T-L tumor tissues and the genome of the case of peripheral blood lymphocytes. The expression of the T-bet (T-box expressed in T cell) gene in the tissue sections of 20 cases of N-NK/T-L, 17 cases of B cell lymphoma (BCL), 3 cases of normal spleen, and 5 cases of chronic nasopharyngitis was detected by in situ hybridization. RESULTS: RLGS combined with VGS revealed a genomic alteration in the T-bet gene and consequent Southern and dot blotting confirmed this genomic alteration in the N-NK/T-L cells. In situ hybridization showed that the T-bet mRNA expression rate of the N-NK/T-L cells was 90.0% (18/20), significantly higher than that of the BCL (11.8%, 2/17, P < 0.01). T-bet mRNA expression was not detected in the normal spleen and chronic nasopharyngitis tissues. CONCLUSION: The amplification and over-expression of the T-bet gene may play a role in the development of N-NK/T-L. The potential value of over-expression of T-bet gene in diagnosis of N-NK/T-L is worth further investigating.