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BACKGROUND: Polycystic ovary syndrome (PCOS) infertility has attracted great attention from researchers due to its high incidence. Numerous studies have shown that Chinese medicine is effective in treating this disease, but there is a wide variety of Chinese medicine therapies available, and there is a lack of comparative evaluation of the efficacy of various Chinese medicine combination therapies in the clinic, which requires further in-depth exploration. This study aims to evaluate the efficacy of a combined traditional Chinese medicine (TCM) therapy for the treatment of infertility with PCOS using network meta-analysis (NMA). METHODS: In PubMed, web of Science, Cochrane Library, Embase, China Knowledge Network, Wanfang Data, VIP Database, China Biomedical Literature Database (SinoMed) databases, searchs were conducted for information about the randomized controlled trials (RCTs) of combined TCM therapy for the treatment of infertility with PCOS. Quality evaluation was performed using the Cochrane 5.3 risk of bias assessment tool, and NMA using Stata 16.0. RESULTS: This study comprised 28 RCTs using 8 combined TCM therapies in total. The results of the NMA showed that moxibustion + herbal, fire acupuncture + herbal, acupuncture + herbal, electroacupuncture + herbal, and acupoint application + herbal improved the clinical pregnancy rate better than acupuncture, herbal, and western medicines monotherapy (P < .05). Additionally, ear point pressure + herbal enema + herbal, acupuncture and moxibustion + herbal, fire acupuncture + herbal, and acupuncture + herbal improved the ovulation rate better than acupuncture, herbal, and western medicines monotherapy (P < .05). Moxibustion + herbal, fire acupuncture + herbal, and acupuncture + herbal are the 3 most effective therapies for improving the clinical pregnancy rate. Fire acupuncture + herbal, acupuncture + herbal, and ear point pressure + herbal enema + herbal are the 3 most effective therapies for improving the ovulation rate. CONCLUSION: The combined TCM therapy demonstrated better efficacy for the treatment of infertility with PCOS compared to acupuncture, herbal, and western medicines monotherapy. However, the optimal treatment therapy varied depending on the outcome indicators. Further large sample, high-quality, and standardized RCTs are needed to verify these findings.

Asunto(s)

Infertilidad Femenina , Medicina Tradicional China , Síndrome del Ovario Poliquístico , Ensayos Clínicos Controlados Aleatorios como Asunto , Humanos , Síndrome del Ovario Poliquístico/complicaciones , Síndrome del Ovario Poliquístico/terapia , Síndrome del Ovario Poliquístico/tratamiento farmacológico , Femenino , Infertilidad Femenina/terapia , Infertilidad Femenina/etiología , Infertilidad Femenina/tratamiento farmacológico , Medicina Tradicional China/métodos , Terapia Combinada , Metaanálisis en Red , Embarazo , Terapia por Acupuntura/métodos , Medicamentos Herbarios Chinos/uso terapéutico , Índice de Embarazo

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OBJECTIVE: Aberrant epigenetic remodeling represents a molecular hallmark in lung adenocarcinoma (LUAD). We aim to investigate the biological roles of SETDB2 and its underlying associations with oxidative stress, providing therapeutic targets for individualized treatment of LUAD. METHODS: Differential analysis was conducted via Limma package, and Kaplan-Meier analysis was performed with survival package. CCK-8, cell proliferation assay, transwell assay, and in vivo assays were conducted to assess the function of SETDB2. Western blot assay, RT-qPCR, and immunohistochemistry (IHC) were conducted to assess the expression levels of SETDB2/NRF2. Chromatin immunoprecipitation (ChIP) assay and ChIP-qPCR were conducted to assess the epigenetic roles of SETDB2. RESULTS: We found that SETDB2 expression is decreased in tumor samples versus normal tissues in TCGA-LUAD cohort, LUAD-EAS cohort, GSE72094 dataset, and independent Soochow-LUAD dataset. Patients with low SETDB2 levels had a worse prognosis relative to those with high SETDB2. SETDB2 inhibition could significantly promote cell growth, migration ability, and stemness maintenance. Gene set enrichment analysis (GSEA) suggested that SETDB2 correlated with oxidative stress crosstalk and regulated NRF2 mRNA levels. ChIP assay suggested that SETDB2 mainly recruited the H3K9me3 enrichment at the NRF2 promoter region to suppress the mRNA levels of NRF2. Downregulated SETDB2 could activate NRF2 transcription and expression, thereby promoting its downstream targets, like NQO1, FTH1, and ME1. Functional experiments demonstrated that low SETDB2 allowed NRF2 to drive malignant processes of LUAD. SETDB2 overexpression attenuated the ability of NRF2 signaling to neutralize cellular reactive oxygen species (ROS) levels, leading to enhanced cell apoptosis. Overexpressed SETDB2 could inhibit tumor progression in vivo and further render LUAD cells sensitive to chemotherapy. CONCLUSIONS: In conclusion, these findings uncovered the suppressive role of SETDB2 in LUAD. SETDB2 negatively regulates NRF2 signaling to modulate tumor progression, which creates a therapeutic vulnerability in LUAD.

Asunto(s)

Adenocarcinoma del Pulmón , Neoplasias Pulmonares , Humanos , Adenocarcinoma del Pulmón/tratamiento farmacológico , Adenocarcinoma del Pulmón/genética , Adenocarcinoma del Pulmón/metabolismo , Proliferación Celular/genética , Regulación Neoplásica de la Expresión Génica , N-Metiltransferasa de Histona-Lisina/genética , N-Metiltransferasa de Histona-Lisina/metabolismo , Neoplasias Pulmonares/tratamiento farmacológico , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/metabolismo , Factor 2 Relacionado con NF-E2/genética , Factor 2 Relacionado con NF-E2/metabolismo , ARN Mensajero

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Lung cancer is the cancer with the highest mortality in the world. So further exploration of the pathogenesis of lung cancer is of great significance. In this study, the specific role and related mechanism of CRIF1 in non-small cell lung cancer (NSCLC) were explored in this research. TheRT-PCR, western blot and IHC assays were used to examine the expression level of CRIF1 in NSCLC tissue, tissue adjacent to carcinoma, NSCLC cell lines and human normal lung epithelial cells. Next, colony formation assay, Alamar blue Kit and EdU assays were employed to examine the proliferation of transfected A549 and NCI-H2009 cells. Measurement of mitochondrial permeability transition pore opening, ATP production and cellular oxygen consumption were used to evaluate the mitochondrial apoptosis of transfected NSCLC cells. Enzymatic activity assays for PYCR1, western blot and flow cytometry assays were used to explore the relationship between PYCR1 and CRIF1. The subcutaneous xenograft tumor mice model was established to explore the role of CRIF1 in vivo. Collectively, results revealed that CRIF1 was upregulated in NSCLC cells and tissues (p < 0.001). CRIF1 promoted proliferation of NSCLC cells (p < 0.001). CRIF1 inhibited mitochondrial apoptosis in NSCLC cells (p < 0.05). Moreover, CRIF1 promoted PYCR1 deacetylation and increased its activity through SIRT3 (p < 0.05). Deacetylation of PYCR1 reversed the antitumor effect of CRIF1 knockdown (p < 0.05). Finally, knockdown of CRIF1 inhibited the tumor growth of NSCLC in vivo (p < 0.05).This research found that CRIF1 promoted the progression of non-small-cell lung cancer by SIRT3- mediated deacetylation of PYCR1.

Asunto(s)

Carcinoma de Pulmón de Células no Pequeñas , Neoplasias Pulmonares , MicroARNs , Sirtuina 3 , Animales , Apoptosis , Carcinoma de Pulmón de Células no Pequeñas/genética , Carcinoma de Pulmón de Células no Pequeñas/patología , Proteínas de Ciclo Celular/metabolismo , Línea Celular Tumoral , Movimiento Celular , Proliferación Celular , Regulación Neoplásica de la Expresión Génica , Humanos , Neoplasias Pulmonares/patología , Ratones , Pirrolina Carboxilato Reductasas , Sirtuina 3/genética , Sirtuina 3/metabolismo , delta-1-Pirrolina-5-Carboxilato Reductasa

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The phenomenon of no-reflow seriously limits the therapeutic value of coronary recanalization and leads to poor prognosis. Recent studies have demonstrated the potential role of pigment epithelium-derived factor (PEDF) in stabilizing endothelial cell junction, reducing vascular permeability and maintaining a quiescent vasculature. In this study, intramyocardial gene delivery was performed 5 days before the acute myocardial infarction/recanalization experiment in male rats. Positron emission tomography perfusion imaging with 13N-NH3 indicated PEDF to promote microvascular reperfusion significantly 4 h postcoronary occlusion. PEDF was observed to maintain the stability of endothelial adherens junctions (AJs), thus preventing the occurrence of no-reflow. PEDF reduced the hypoxia-induced vascular endothelial (VE)-cadherin endocytosis through PEDF/LR/Src/VE-cadherin S665 axis in vitro, which was remarkably observed to maintain endothelial AJs. Generally, PEDF might function as a relevant target for therapeutic vasculoprotection by way of regulating the phosphorylation level of VE-cadherin according to our data, thus being crucial for preventing no-reflow.

Asunto(s)

Infarto del Miocardio , Serpinas , Animales , Proteínas del Ojo/genética , Masculino , Infarto del Miocardio/terapia , Factores de Crecimiento Nervioso/genética , Ratas , Serpinas/genética , Serpinas/farmacología

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INTRODUCTION: The literature regarding the application of uniportal video-assisted thoracoscopic segmental resection of the lung in patients aged over 65 years with non-small cell lung cancer (NSCLC) is sparse. This paper reports 175 cases of uniportal video-assisted thoracoscopic segmental resection of the lung performed at one center, of which 63 patients were over 65 years old. AIM: To investigate the safety and feasibility of uniportal video-assisted thoracoscopic segmental resection of the lung in elderly patients aged over 65 years with NSCLC. MATERIAL AND METHODS: A retrospective analysis of 175 NSCLC patients who underwent uniport video-assisted thoracoscopic segmental resection of the lung in the center from August 2018 to August 2020 was conducted, and based on the age of 65 years, patients were divided into elderly and non-elderly groups. The general data and perioperative indicators of the two groups were compared. RESULTS: The procedures were completed in all patients without death or conversion to open surgery. In the general data of the two groups of patients, the prevalence of emphysema in the elderly group was significantly higher than that in the non-elderly group (p = 0.001). However, there was no statistically significant difference between the two groups in surgery time, intraoperative blood loss, thoracic drainage tube retention time, postoperative hospital stay, incision satisfaction, or postoperative complications (p > 0.05). CONCLUSIONS: Uniportal video-assisted thoracoscopic segmental resection of the lung is feasible and safe in elderly patients with NSCLC aged over 65 years.

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OBJECTIVE: To investigate the clinical significance of cyclin-dependent kinase 14 (CDK14) expression in patients with non-small cell lung cancer (NSCLC). METHODS: The present prospective observational study included 193 patients diagnosed with NSCLC between January 2010 and December 2014. NSCLC tumor tissues and paired paracancerous normal tissues were obtained from all patients. CDK14, thyroid transcription factor 1 (TTF-1), cytokeratin 5/6 (CK5/6), and Ki67 expression was measured via immunohistochemistry (IHC). RESULTS: CDK14 staining was strong (>3) in 129 patients (66.49%) and weak (≤3) in 64 patients (33.16%). The mean IHC scores were markedly higher in tumor tissues than in paracancerous tissues. Pearson's analysis demonstrated that the IHC scores of CDK14 expression were positively correlated with TTF-1, CK5/6, and Ki67 scores. Kaplan-Meier analysis illustrated that 5-year overall survival was markedly longer in patients with weak CDK14 staining. TNM stage, pleural invasion, lymph node metastasis, CDK14 expression, and Ki67 expression were risk factors for 5-year overall survival in patients with NSCLC. CONCLUSION: CDK14 overexpression portended poor outcomes in patients with NSCLC, and CDK14 expression was correlated with TTF-1, CK5/5, and Ki67 expression.

Asunto(s)

Carcinoma de Pulmón de Células no Pequeñas , Neoplasias Pulmonares , Carcinoma de Pulmón de Células no Pequeñas/diagnóstico , Carcinoma de Pulmón de Células no Pequeñas/genética , Quinasas Ciclina-Dependientes , Humanos , Inmunohistoquímica , Neoplasias Pulmonares/diagnóstico , Neoplasias Pulmonares/genética , Metástasis Linfática , Pronóstico

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MicroRNAs (miRNAs/miRs) in extracellular vesicles (EVs) are potential diagnostic markers. The purpose of the present study was to investigate potential EV miRNA biomarkers for lung adenocarcinoma (LUAD). Potential miRNAs were identified by searching public databases and verified by examining clinical samples. The diagnostic value of EV-associated miR-10b, plasma miR-10b and tumor markers (TMs), including α-fetoprotein (AFP), neuron-specific enolase, carcinoembryonic antigen (CEA), cytokeratin 19 fragment 21-1 (CYFRA211), pro-gastrin-releasing-peptide, carbohydrate antigen (CA)125, CA153, CA199 and CA724, was evaluated via receiver operating characteristic curve analysis. By searching the Gene Expression Omnibus and The Cancer Genome Atlas databases, miR-10b was identified as a potential biomarker. The analysis of clinical samples suggested that EV-associated miR-10b from plasma was significantly differentially expressed between LUAD and control samples. EV-associated miR-10b could function as a diagnostic marker for LUAD, with an AUC of 0.998, which was higher than the AUCs for TMs such as AFP, CEA, CYFRA211, CA125, CA153, CA199, CA724, pro-gastrin-releasing-peptide and neuron-specific enolase. In conclusion, EV-associated miR-10b may be a potential diagnostic biomarker for LUAD that is superior to plasma miR-10b and TMs.

RESUMEN

To explore the molecular mechanism in the esophageal squamous carcinoma (ESCC) environment, we selected datasets of ESCC patients from The Cancer Genome Atlas (TCGA) (n = 78) and explored the infiltration condition of 24 immune cells in each sample. We assorted the microenvironment of ESCC into two Infiltration groups (I and II) and built a random forest classifier model. We showed traits of gene and clinicopathology in the tumor microenvironment (TME) phenotypes systematically. Infiltration I had low infiltration of immune cells and immunomodulators but relatively higher mutation load, while Infiltration II was enriched with cytotoxic T cells and immunosuppressive cells. The upregulation of several immune cytokines like IFN-γ, TNF-ß, and PD-L1 was seen in Infiltration II. The infiltration group was an independent predictor of prognosis showed by Multivariable Cox analysis (Infiltration II vs. I, hazard ratio = 2.73, 95% confidence interval = 1.08-6.91, p = 0.03). All the results can be verified in datasets from the Gene Expression Omnibus database (GEO) and our institution (n = 98). Our results demonstrate a synthesis of the infiltration pattern of the immune in ESCC. We reveal the mechanism of TME, which may contribute to the progress of immunotherapy for patients with ESCC.

Asunto(s)

Neoplasias Esofágicas/genética , Neoplasias Esofágicas/inmunología , Carcinoma de Células Escamosas de Esófago/genética , Carcinoma de Células Escamosas de Esófago/inmunología , Anciano , Citocinas/genética , Citocinas/inmunología , Neoplasias Esofágicas/mortalidad , Carcinoma de Células Escamosas de Esófago/mortalidad , Femenino , Regulación Neoplásica de la Expresión Génica , Genoma , Humanos , Masculino , Pronóstico , Linfocitos T/inmunología , Microambiente Tumoral/genética , Microambiente Tumoral/inmunología

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Malignant tumor represents a major reason for death in the world and its incidence is growing rapidly. Developing the tools for early diagnosis is possibly a promising way to offer diverse therapeutic options and promote the survival chance. Secreted phosphoprotein 1 (SPP1), also called Osteopontin (OPN), has been demonstrated overexpressed in many cancers. However, the specific role of SPP1 in prognosis, gene mutations, and changes in gene and miRNA expression in human cancers is unclear. In this report, we found SPP1 expression was higher in most of the human cancers. Based on Kaplan-Meier plotter and the PrognoScan database, we found high SPP1 expression was significantly correlated with poor survival in various cancers. Using a large dataset of colon adenocarcinoma (COAD), head and neck cancer (HNSC), lung adenocarcinoma (LUAD), and lung squamous cell carcinoma (LUSC) patients from the Gene Expression Omnibus (GEO) and The Cancer Genome Atlas (TCGA) databases, this study identified 22 common genes and 2 common miRNAs. GO, and KEGG paths analyses suggested that SPP1 correlated genes were mainly involved in positive regulation of immune cell activation and infiltration. SPP1-associated genes and miRNAs regulatory networks suggested that their interactions may play a role in the progression of four selected cancers. SPP1 showed significant positive correlation with the immunocyte and immune marker sets infiltrating degrees. All of these data provide strong evidence that SPP1 may promote tumor progress through interacting with carcinogenic genes and facilitating immune cells' infiltration in COAD, HNSC, LUAD, and LUSC.

RESUMEN

INTRODUCTION: Thoracoscopic sympathectomy (TS) has been proven to be a safe and effective treatment for primary palmar hyperhidrosis (PH). However, the complications include compensatory hyperhidrosis (CH), and over-dry hands may occur in some patients after TS. AIM: To compare the therapeutic effect of T3 and T4 TS on primary PH and primary PH with axillary and plantar sweating. MATERIAL AND METHODS: We retrospectively analyzed 100 patients with PH who had undergone T3 (group A, n = 49) or T4 (group B, n = 51) TS in our department, with at least 1 year of postoperative follow-up. RESULTS: At discharge, no major complications or deaths occurred in either group. The condition of sweaty hands was fully improved in 44 of 49 patients in group A and all patients in group B, with a significant difference (p = 0.031). After 12 months of follow-up, 18 (36.7%) patients in group A and 4 (7.8%) patients in group B developed CH, 16 (48.5%) patients in group A and 24 (77.4%) patients in group B had improved axillary sweating, with a significant difference (p < 0.05). The satisfaction rate of group B was significantly higher than that of group A (p < 0.01). CONCLUSIONS: Both T3 and T4 TS were safe and effective treatments for PH patients, but the incidence of CH in T4 TS was lower than that in T3 TS. T3 TS may be more suitable for patients with severe PH, while T4 TS had a better therapeutic effect on PH patients with axillary sweating.

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The application of cancer chronotherapy is to treat cancers based on at specific times during circadian rhythms. Previous studies have characterized the impact of circadian clock on tumorigenesis and specific immune cells. Here, by using multi-omics computation techniques, we systematically characterized the distinct roles of core circadian clock genes in thoracic cancers including lung adenocarcinoma, lung squamous cell carcinoma, and esophageal carcinoma. Strikingly, a wide range of core clock genes are epigenetically altered in lung adenocarcinomas and lung squamous cell carcinomas but not esophageal carcinomas. Further cancer hallmark analysis reveals that several core clock genes highly correlate with apoptosis and cell cycle such as RORA and PER2. Interestingly, our results reveal that CD4 and CD8 T cells are correlated with core clock molecules especially in lung adenocarcinomas and lung squamous cell carcinomas, indicating that chrono-immunotherapy may serve as a candidate option for future cancer management.

Asunto(s)

Carcinoma de Pulmón de Células no Pequeñas/genética , Relojes Circadianos/genética , Neoplasias Esofágicas/genética , Neoplasias Pulmonares/genética , Microambiente Tumoral/fisiología , Carcinoma de Pulmón de Células no Pequeñas/inmunología , Carcinoma de Pulmón de Células no Pequeñas/patología , Metilación de ADN , Neoplasias Esofágicas/inmunología , Neoplasias Esofágicas/patología , Humanos , Neoplasias Pulmonares/inmunología , Neoplasias Pulmonares/patología , Transcriptoma/genética

RESUMEN

Circulating tumor cells (CTCs) are an independent prognostic marker in non-small cell lung cancer (NSCLC). CTC numbers are closely related to early diagnosis, clinical stage, therapy surveillance, and prognosis of NSCLC. We used a more efficient nano-enrichment method to detect CTCs in NSCLC patients and explored the clinical value of CTCs. The results showed that CTC numbers in stage IV cases were significantly higher than those in stage I, II or III cases. The number of CTCs in poorly-differentiated cases was significantly higher than that in well-differentiated cases. During six chemotherapy cycles, the average CTC number decreased from 5.8/7.5 ml in cycle #1 to 2.4/7.5 ml in cycle #4 and remained at almost the same level from 4 to 6 cycles. CTC numbers in patients with EGFR mutations was significantly higher than those in patients with no mutations. The average progression free survival (PFS) in the favorable group (CTC ≤ 5/7.5 ml) was 11.3 months, which was longer than that in the unfavorable group (CTC > 5/7.5 ml, 7.2 months). In conclusion, the assessment of NSCLC cannot be performed using a single CTC analysis. The clinical value is more significant in the continuous analysis of CTC data, as well as the cross-validation of other indexes and imaging results.

Asunto(s)

Carcinoma de Pulmón de Células no Pequeñas/patología , Neoplasias Pulmonares/patología , Células Neoplásicas Circulantes/patología , Adulto , Anciano , Anciano de 80 o más Años , Carcinoma de Pulmón de Células no Pequeñas/tratamiento farmacológico , Carcinoma de Pulmón de Células no Pequeñas/genética , Recuento de Células , Diferenciación Celular , Separación Celular/métodos , Receptores ErbB/genética , Femenino , Humanos , Neoplasias Pulmonares/tratamiento farmacológico , Neoplasias Pulmonares/genética , Masculino , Persona de Mediana Edad , Mutación , Nanotecnología/métodos , Estadificación de Neoplasias , Células Neoplásicas Circulantes/efectos de los fármacos , Pronóstico , Supervivencia sin Progresión

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BACKGROUND: This study analyzed the clinical features and prognosis of basaloid squamous cell carcinoma of the lung (BSC), and constructed a nomogram to predict the prognoses of patients. METHODS: The information of pure BSC patients was obtained in the Surveillance, Epidemiology, and End Results database between 2004 and 2015. Then, it was evaluated, and compared with the data of lung squamous cell carcinoma (SCC), lung large cell carcinoma (LCC) and lung adenocarcinoma (LAC) patients. Subsequently, we used univariate and multivariate analyses to investigate the independent factors related to the prognoses of patients with BSC and constructed a nomogram to verify the prognoses. RESULTS: A total of 425 patients diagnosed with BSC were enrolled. Compared with patients with SCC, LCC and LAC, the mean survival time of BSC patients was better than all of them. Compared with SCC, there were significant differences between the characteristics of grade (P < 0.001), total stage (P < 0.001), T stage (P < 0.001), N stage (P < 0.001), M stage (P < 0.001), surgery (P < 0.001), radiotherapy (P < 0.001), and chemotherapy (P < 0.001), while BSC also had significantly different clinical characteristics from LCC and LAC. Univariate and multivariate survival analyses showed that age (P < 0.001), T stage (P < 0.001), N stage (P = 0.009), M stage (P < 0.001), and surgery (P < 0.001) were independent prognostic factors of BSC. The survival of patients undergoing lobectomy was significantly better than sublobar resection, with an OR of 0.389 (0.263-0.578). We constructed a nomogram with a C-index of 0.750 (95% confidence interval) based on the results of multivariate analysis. The calibration curves based on nomogram scores indicated that the nomogram could accurately predict the prognosis of patients. CONCLUSIONS: BSC had unique clinical and prognostic features. T stage, N stage, M stage, age, and surgery were independently associated with overall survival (OS). Lobectomy was a relative ideal choice for patients with BSC. The nomogram effectively predicted the OS at 1-, 3-, and 5-years.

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BACKGROUND: Esophageal large cell neuroendocrine carcinoma (ELCNC) seems a rarely gastrointestinal malignancy. By far, its clinicopathological characteristics and prognosis have not been deeply studied. METHODS: The data of patients having ELCNC was extracted from the Surveillance, Epidemiology, and End Results (SEER) database, then assessed and compared with information from patients with esophageal small cell neuroendocrine carcinoma (ESCNC) or esophageal squamous cell carcinoma (ESCC). We used univariate and multivariate analyses to accurately detect independent prognostic factors. RESULTS: The data of 36 patients for ELCNC were obtained between 2004 and 2015. Compared with patients with ESCNC and ESCC, the mean survival time of ECLNC patients was worse than those with ESCC, while similar to ESCNC. Thus, ELCNC had significantly different clinicopathological characteristics compared to ESCNC and ESCC. Univariate and multivariate analyses revealed that age (P=0.001) and M stage (P=0.004) were independent prognostic factors. CONCLUSIONS: ELCNC is a rare subtype of esophageal neuroendocrine carcinoma. The clinicopathological features differ from those of other esophageal carcinomas. Prognosis may be closely related to age and M stage.

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Angiogenesis assays are important tools for studying both the mechanisms of cardiac angiogenesis and the potential development of therapeutic strategies to ischemic heart diseases. Currently, various assays have been used to quantitate cardiac tubule formation, yet no consensus has been reached regarding a suitable assay for evaluating the efficacy of angiogenic stimulants or inhibitors. Most in vivo angiogenesis assays are complex and difficult to interpret, whereas traditional in vitro angiogenesis models measure only one aspect of this process. To bridge the gap between in vivo and in vitro angiogenesis assays, here, we have developed a novel modified cardiac explants matrigel assay. We observed the morphology of vascular sprouts formed in three forms of cardiac angiogenesis assays then used quantitative image analyses to further compare the morphological features of vascular sprouts formed in two cardiac explants angiogenesis assays. Vascular sprouts formed in the fibronectin group were less and short, whereas those formed in the matrigel group were significantly longer, consisting of more area and branch points. Moreover, we found the benefits of this matrigel model by observing the ability of cardiac explants to form vascular sprouts under normoxia or hypoxia condition in the presence of angiogenic stimulant and inhibitor, VEGF and PEDF. In summary, the above analyses revealed that the morphology of vascular sprouts formed in this model appears more representative of myocardial capillary formation in vivo, and this accessible, reliable angiogenic assay is a more physiologically relevant assay which allows further assessment of pharmacologic compounds on cardiac angiogenesis.

Asunto(s)

Vasos Coronarios/fisiología , Neovascularización Fisiológica , Inductores de la Angiogénesis/farmacología , Inhibidores de la Angiogénesis/farmacología , Animales , Animales Recién Nacidos , Hipoxia de la Célula , Microambiente Celular , Colágeno/metabolismo , Vasos Coronarios/efectos de los fármacos , Vasos Coronarios/metabolismo , Combinación de Medicamentos , Fibronectinas/metabolismo , Células Endoteliales de la Vena Umbilical Humana/efectos de los fármacos , Células Endoteliales de la Vena Umbilical Humana/fisiología , Humanos , Laminina/metabolismo , Neovascularización Fisiológica/efectos de los fármacos , Proteoglicanos/metabolismo , Ratas Sprague-Dawley , Factores de Tiempo , Técnicas de Cultivo de Tejidos

RESUMEN

Endothelial mesenchymal transition (EndMT) plays a critical role in the pathogenesis and progression of interstitial and perivascular fibrosis after acute myocardial infarction (AMI). Pigment epithelium-derived factor (PEDF) is shown to be a new therapeutic target owing to its protective role in cardiovascular disease. In this study, we tested the hypothesis that PEDF is an endogenous inhibitor of EndMT and represented a novel mechanism for its protective effects against overactive cardiac fibrosis after AMI. Masson's trichrome (MTC) staining and picrosirius red staining revealed decreased interstitial and perivascular fibrosis in rats overexpressing PEDF. The protective effect of PEDF against EndMT was confirmed by co-labeling of cells with the myofibroblast and endothelial cell markers. In the endothelial cells of microvessels in the ischemic myocardium, the inhibitory effect of PEDF against nuclear translocation of ß-catenin was observed through confocal microscopic imaging. The correlation between antifibrotic effect of PEDF and inactivation of ß-catenin was confirmed by co-transfecting cells with lentivirus carrying PEDF or PEDF RNAi and plasmids harboring ß-catenin siRNA(r) or constitutive activation of mutant ß-catenin. Taken together, these results establish a novel finding that PEDF could inhibit EndMT related cardiac fibrosis after AMI by a mechanism dependent on disruption of ß-catenin activation and translocation.

Asunto(s)

Cardiomiopatías/prevención & control , Endotelio Vascular/citología , Transición Epitelial-Mesenquimal , Proteínas del Ojo/metabolismo , Fibrosis/prevención & control , Infarto del Miocardio/complicaciones , Miocardio/metabolismo , Factores de Crecimiento Nervioso/metabolismo , Serpinas/metabolismo , Enfermedad Aguda , Animales , Apoptosis , Cardiomiopatías/etiología , Movimiento Celular , Células Cultivadas , Endotelio Vascular/metabolismo , Proteínas del Ojo/administración & dosificación , Proteínas del Ojo/genética , Fibrosis/etiología , Masculino , Factores de Crecimiento Nervioso/administración & dosificación , Factores de Crecimiento Nervioso/genética , Ratas , Ratas Sprague-Dawley , Serpinas/administración & dosificación , Serpinas/genética , Transducción de Señal

RESUMEN

Pigment epithelial-derived factor (PEDF) is a glycoprotein with broad biological activities including inhibiting oxygen-glucose deprivation(OGD)-induced cardiomyocytes apoptosis through its anti-oxidative properties. PEDF derived peptide-44mer shows similar cytoprotective effect to PEDF. However, the molecular mechanisms mediating cardiomyocytes apoptosis have not been fully established. Here we found that PEDF and 44mer decreased the content of ROS. This content was abolished by either PEDF-R small interfering RNA (siRNA) or PPARγ antagonist. The level of Lysophosphatidic acid (LPA) and phospholipase A2 (PLA2) was observed as drawn from the ELISA assays. PEDF and 44mer sequentially induced PPARγ expression was observed both in qPCR and Western blot assays. The level of LPA and PLA2 and PPARγ expression increased by PEDF and 44mer was significantly attenuated by PEDF-R siRNA. However, PEDF and 44mer inhibited the H9c2 cells and cultured neonatal rat myocardial cells apoptosis rate. On the other hand, TUNEL assay and cleavage of procaspase-3 showed that PEDF-R siRNA or PPARγ antagonist increased the apoptosis again. We conclude that under OGD condition, PEDF and 44mer reduce H9c2 cells apoptosis and inhibit OGD-induced oxidative stress via its receptor PEDF-R and the PPARγ signaling pathway.

Asunto(s)

Proteínas del Ojo/metabolismo , Miocitos Cardíacos/metabolismo , Factores de Crecimiento Nervioso/metabolismo , Estrés Oxidativo/fisiología , PPAR gamma/metabolismo , Péptidos/metabolismo , Receptores de Neuropéptido/metabolismo , Serpinas/metabolismo , Línea Celular , Glucosa/metabolismo , Humanos , Peso Molecular , Miocitos Cardíacos/citología , Oxígeno/metabolismo , Péptidos/química , Regulación hacia Arriba/fisiología

RESUMEN

Pigment epithelial-derived factor (PEDF) is a potent anti-angiogenic factor whose effects are partially mediated through the induction of endothelial cell apoptosis. However, the underlying mechanism for PEDF and the functional PEDF peptides 34-mer and 44-mer to inhibit angiogenesis in the heart has not been fully established. In the present study, by constructing adult Sprague-Dawley rat models of acute myocardial infarction (AMI) and in vitro myocardial angiogenesis, we showed that PEDF and 34-mer markedly inhibits angiogenesis by selectively inducing tip cells apoptosis rather than quiescent cells. Peptide 44-mer on the other hand exhibits no such effects. Next, we identified Fas death pathway as essential downstream regulators of PEDF and 34-mer activities in inhibiting angiogenesis. By using peroxisome proliferator-activated receptor γ (PPAR-γ) siRNA and PPAR-γ inhibitor, GW9662, we found the effects of PEDF and 34-mer were extensively blocked. These data suggest that PEDF and 34-mer inhibit angiogenesis via inducing tip cells apoptosis at least by means of up-regulating PPAR-γ to increase surface FasL in the ischemic heart, which might be a novel mechanism to understanding cardiac angiogenesis after AMI.

Asunto(s)

Proteínas del Ojo/farmacología , Proteína Ligando Fas/genética , Infarto del Miocardio/genética , Neovascularización Fisiológica/efectos de los fármacos , Factores de Crecimiento Nervioso/farmacología , PPAR gamma/genética , Péptidos/farmacología , Serpinas/farmacología , Secuencia de Aminoácidos , Anilidas/farmacología , Animales , Apoptosis/efectos de los fármacos , Células Endoteliales/citología , Células Endoteliales/efectos de los fármacos , Células Endoteliales/metabolismo , Proteínas del Ojo/genética , Proteínas del Ojo/metabolismo , Proteína Ligando Fas/agonistas , Proteína Ligando Fas/metabolismo , Regulación de la Expresión Génica , Corazón/efectos de los fármacos , Corazón/fisiopatología , Datos de Secuencia Molecular , Infarto del Miocardio/metabolismo , Infarto del Miocardio/patología , Factores de Crecimiento Nervioso/genética , Factores de Crecimiento Nervioso/metabolismo , PPAR gamma/agonistas , PPAR gamma/antagonistas & inhibidores , PPAR gamma/metabolismo , Péptidos/síntesis química , Cultivo Primario de Células , ARN Interferente Pequeño/genética , ARN Interferente Pequeño/metabolismo , Ratas , Ratas Sprague-Dawley , Serpinas/genética , Serpinas/metabolismo , Transducción de Señal

RESUMEN

BACKGROUND: Pigment epithelium-derived factor (PEDF) is a 50-kDa secreted glycoprotein that is highly expressed in cardiomyocytes. A variety of peptides derived from PEDF exerts diverse physiological activities including anti-angiogenesis, antivasopermeability, and neurotrophic activities. Recent studies demonstrated that segmental functional peptides of PEDF, 44mer peptide (Val78-Thr121), show similar neurotrophic and cytoprotective effect to that of the holoprotein. We found that PEDF can reduce infarct size and protect cardiac function after acute myocardial infarction (AMI). However, the effects of PEDF on cardiac triglyceride (TG) accumulation after AMI remain unknown. The present study was performed to demonstrate the influence of PEDF and its functional peptides 44mer on TG degradation in AMI. METHODS: The left ascending coronary artery (LAD) was ligated to induce AMI. PEDF-small interfering RNA (siRNA)-lentivirus (PEDF-RNAi-LV) or PEDF-LV was delivered to the ischemic myocardium in order to knock down or overexpress PEDF, respectively. Oil Red O staining and a TG assay kit were used to analyze the TG content in cardiomyocytes and infarcted areas. RESULTS: The TG content significantly decreased in the PEDF-overexpressing heart compared to the sham group (P < 0.05). Both rPEDF and 44mer administration stimulate the TG degradation in cultured cardiomyocytes (P < 0.05). Adipose triglyceride lipase (ATGL)-specific inhibitor, atglistatin, attenuated the PEDF or 44mer-induced TG lipolysis activation of cardiomyocytes at 10 µmol/L. The effects of PEDF and 44mer on myocardial TG degradation were also abolished when ATGL was downregulated. CONCLUSIONS: We conclude that PEDF and 44mer promote TG degradation in cardiomyocytes after AMI via ATGL. The substitution of PEDF and 44mer may be a novel therapeutic strategy for cardiac TG accumulation after AMI.

Asunto(s)

Proteínas del Ojo/metabolismo , Lipasa/metabolismo , Miocardio/metabolismo , Factores de Crecimiento Nervioso/metabolismo , Péptidos/metabolismo , Serpinas/metabolismo , Triglicéridos/metabolismo , Aciltransferasas/genética , Aciltransferasas/metabolismo , Secuencia de Aminoácidos , Animales , Hipoxia de la Célula , Gotas Lipídicas/metabolismo , Lipólisis , Masculino , Datos de Secuencia Molecular , Infarto del Miocardio/complicaciones , Infarto del Miocardio/patología , Isquemia Miocárdica/complicaciones , Isquemia Miocárdica/patología , Miocardio/ultraestructura , Miocitos Cardíacos/metabolismo , Miocitos Cardíacos/patología , Péptidos/química , ARN Mensajero/genética , ARN Mensajero/metabolismo , Ratas Sprague-Dawley