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OBJECTIVE: Radiation for locally advanced esophageal squamous cell carcinoma often is accompanied by radiation esophagitis, which interferes with oral intake. We aimed to develop a nomogram model to identify initially inoperable patients with relative and absolute weight loss who need prophylactic nutritional supplementation. METHODS: A total of 365 initially inoperable patients with locally advanced esophageal squamous cell carcinoma receiving radiotherapy between January 2018 and December 2022 were included in the study, which was divided into discovery and validation cohorts. Receiver operating characteristic and Kaplan-Meier curve analyses were performed to compare the areas under the curve and survival benefits. RESULTS: A total of 42.2% (154 of 365) of the patients had been diagnosed with cancer cachexia. The malnourished group had a higher interruption rate of radiotherapy and number of complication diseases (P < 0.05). Meanwhile, patients with malnutrition had lower lymphocytes and prognostic nutritional index (P < 0.05). The combined index showed a higher area under the curve value (0.67; P < 0.001) than number of complication diseases (area under the curve = 0.52) and prognostic nutritional index (area under the curve = 0.49) for relative weight loss (≥ 5%). Similarly, the combined index had a higher area under the curve value (0.79; P < 0.001) than number of complication diseases (area under the curve = 0.56), treatment regimens (area under the curve = 0.56), subcutaneous fat thickness (area under the curve = 0.60), pretreatment body weight (area under the curve = 0.61), neutrophils (area under the curve = 0.56), and prognostic nutritional index (area under the curve = 0.50) for absolute weight loss (≥ 5 kg). Absolute and relative weight loss remained independent prognostic factors, with short overall survival rates compared with the normal group (P < 0.05). Patients with high nomogram scores supported by nutritional intervention had less weight loss, better nutrition scores, and increased plasma CD8+ T cells, and interferon gamma. CONCLUSIONS: We developed a nomogram model that was intended to estimate relative and absolute weight loss in initially inoperable patients with locally advanced esophageal squamous cell carcinoma during radiotherapy, which might help facilitate an objective decision on prophylactic nutritional supplementation.
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Carcinoma de Células Escamosas , Neoplasias Esofágicas , Carcinoma de Células Escamosas de Esófago , Humanos , Carcinoma de Células Escamosas de Esófago/radioterapia , Nomogramas , Neoplasias Esofágicas/complicaciones , Neoplasias Esofágicas/radioterapia , Carcinoma de Células Escamosas/radioterapia , Carcinoma de Células Escamosas/tratamiento farmacológico , Pérdida de PesoRESUMEN
BACKGROUND: Intraductal papillary neoplasm of the bile duct (IPNB) is a rare distinct subtype of precursor lesions of biliary carcinoma. IPNB is considered to originate from luminal biliary epithelial cells, typically displays mucin-hypersecretion or a papillary growth pattern, and results in cystic dilatation[1]. IPNB develops anywhere in the intrahepatic and extrahepatic biliary tracts, and can occur in various pathological stages from low-grade dysplasia to invasive carcinoma. IPNBs have similar phenotypic changes in the occurrence and development of all subtypes, and the prognosis is significantly better than that of traditional (non-papillary) cholangiocarcinoma. AIM: To evaluate the clinicopathological features of IPNB to provide evidence-based guidance for treatment. METHODS: Invasive IPNB, invasive intraductal papillary mucinous neoplasm of the pancreas (IPMN), and traditional cholangiocarcinoma data for affected individuals from 1975 to 2016 were obtained from the Surveillance, Epidemiology, and End Results (SEER) database. Annual percentage changes (APCs) in the incidence and incidence-based (IB) mortality were calculated. We identified the independent predictors of overall survival (OS) and cancer-specific survival (CSS) in individuals with invasive IPNB. RESULTS: The incidence and IB mortality of invasive IPNB showed sustained decreases, with an APC of -4.5% (95%CI: -5.1% to -3.8%) and -3.3% (95%CI: -4.1% to -2.6%) (P < 0.001), respectively. Similar decreases in incidence and IB mortality were seen for invasive IPMN but not for traditional cholangiocarcinoma. Both OS and CSS for invasive IPNB were better than for invasive IPMN and traditional cholangiocarcinoma. A total of 1635 individuals with invasive IPNB were included in our prognosis analysis. The most common tumor sites were the pancreaticobiliary ampulla (47.9%) and perihilar tract (36.7%), but the mucin-related subtype of invasive IPNB was the main type, intrahepatically (approximately 90%). In the univariate and multivariate Cox regression analysis, age, tumor site, grade and stage, subtype, surgery, and chemotherapy were associated with OS and CSS (P < 0.05). CONCLUSION: Incidence and IB mortality of invasive IPNB trended steadily downward. The heterogeneity of IPNB comprises site and the tumor's mucin-producing status.
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BACKGROUND: N1-methyladenosine (m1A) is a reversible post-transcriptional modification in mRNA, which has been proved to play critical roles in various biological processes through interaction with different m1A regulators. There are several m1A regulators existing in the human genome, including YTHDF1-3 and YTHDC1. METHODS: Several techniques have been developed to identify the substrates of m1A regulators, but their binding specificity and biological functions are not yet fully understood due to the limitations of wet-lab approaches. Here, we submitted the framework m1ARegpred (m1A regulators substrate prediction), which is based on machine learning and the combination of sequence-derived and genome-derived features. RESULTS: Our framework achieved area under the receiver operating characteristic (AUROC) scores of 0.92 in the full transcript model and 0.857 in the mature mRNA model, showing an improvement compared to the existing sequence-derived methods. In addition, motif search and gene ontology enrichment analysis were performed to explore the biological functions of each m1A regulator. CONCLUSIONS: Our work may facilitate the discovery of m1A regulators substrates of interest, and thereby provide new opportunities to understand their roles in human bodies.
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Adenosina , Genómica , Adenosina/genética , Adenosina/metabolismo , Humanos , ARN Mensajero/genéticaRESUMEN
Tripterygium wilfordii Hook F. is a well-known but poisonous traditional Chinese medicine used for treating a wide variety of inflammatory and autoimmune disorders. Celastrol, a quinone methyl triterpenoid compound and a representative component of T. wilfordii Hook F., shows a variety of pharmacological activities, such as anti-inflammatory and antitumor activities. Here, we investigated the antineuropathic pain (NP) effect of celastrol and its potential mechanisms. Rats with chronic constrictive injury (CCI)-induced NP were used to evaluate the analgesic effect of celastrol. Gabapentin was used as a reference compound (positive control). The results showed that gabapentin (100 mg/kg, i.p.) and multiple doses of celastrol (0.5, 1 and 2 mg/kg, i.p.) increased the threshold of mechanical and thermal pain in the rats with NP. Western blot results showed that celastrol significantly inhibited the activation of microglia and astrocytes in the spinal cord of rats with NP. Additionally, the levels of the proinflammatory cytokines tumor necrosis factor α (TNF-α), interleukin 1ß and interleukin 6, detected by ELISA in the spinal cord of the rats with NP, were significantly inhibited by celastrol. Furthermore, celastrol treatment dramatically inhibited the expression of the TLR4/NF-κB signaling pathway in the spinal cord. Taken together, our findings suggested that celastrol could attenuate mechanical and thermal pain in CCI-induced NP, and this protection might be attributed to inhibiting the TLR4/NF-κB signaling pathway and exerting anti-inflammatory effects in the spinal cord.
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FN-kappa B , Neuralgia , Animales , FN-kappa B/metabolismo , Neuralgia/tratamiento farmacológico , Triterpenos Pentacíclicos , Ratas , Ratas Sprague-Dawley , Transducción de Señal , Médula Espinal/metabolismo , Receptor Toll-Like 4/genéticaRESUMEN
Evidence has documented the tumor-promoting properties of long non-coding RNA (lncRNA) FOXD2 adjacent opposite strand RNA 1 (FOXD2-AS1) in many cancers. However, little is known about its role in gallbladder cancer. Here, we aimed to characterize the functional relevance of lncRNA FOXD2-AS1 in gallbladder cancer and the possible mechanisms associated with methylation of MutL homolog-1 (MLH1). Initially, microarray-based gene expression profiling of gallbladder cancer was employed to identify differentially expressed lncRNAs. Next, the expression of lncRNA FOXD2-AS1 was examined, and the cell line presenting with the highest lncRNA FOXD2-AS1 expression was selected for subsequent experimentation. Then, the interaction between lncRNA FOXD2-AS1 and MLH1 was identified. The effect of lncRNA FOXD2-AS1 on proliferation, migration, invasion, and apoptosis as well as tumorigenicity of transfected GBC-SD cells was examined with gain- and loss-of-function experiments. We found that lncRNA FOXD2-AS1 was highly expressed, while MLH1 was poorly expressed in gallbladder cancer cells. Besides, lncRNA FOXD2-AS1 could promote MLH1 methylation by recruiting DNMT1 to the MLH1 promoter, and consequently inhibit MLH1 transcription. Silencing of lncRNA FOXD2-AS1 or overexpression of MLH1 inhibited gallbladder cancer cell proliferation, invasion, and migration, while facilitating cell apoptosis in vitro as well as retarding tumor growth in vivo. Thus, silencing of lncRNA FOXD2-AS1 suppressed the progression of gallbladder cancer via upregulation of MLH1 by inhibiting MLH1 promoter methylation. These findings present lncRNA FOXD2-AS1 knockdown as a potential candidate for the treatment of gallbladder cancer.
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Neoplasias de la Vesícula Biliar , Homólogo 1 de la Proteína MutL , ARN Largo no Codificante , Línea Celular Tumoral , Proliferación Celular/genética , Neoplasias de la Vesícula Biliar/genética , Neoplasias de la Vesícula Biliar/terapia , Regulación Neoplásica de la Expresión Génica , Silenciador del Gen , Humanos , Metilación , Homólogo 1 de la Proteína MutL/genética , ARN Largo no Codificante/genéticaRESUMEN
Twenty-four patients with cold, dampness, obstructive ankylosing spondylitis were treated with sulfasalazine and sulfasalazine in combination with moxibustion for 3 weeks. The results showed that the combined treatment of traditional Chinese and western medicine was significantly higher than those of western medicine treatment, meanwhile, the scoreofsymptoms quantification, C-reactive protein and erythrocyte sedimentation rate of the integrated Chinese and western medicine treatment were significantly lower than those of western medicine treatment, and the level of physical signs was significantly higher than that of western medicine treatment, and there were no significant differences in adverse reactions. Moxibustion combined with sulfasalazine in the treatment of cold and damp obstructive ankylosing spondylitis can effectively improve the characteristics of the body, relieve pain symptoms and improve the prognosis.
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Antiinflamatorios no Esteroideos/uso terapéutico , Moxibustión , Espondilitis Anquilosante/terapia , Sulfasalazina/uso terapéutico , Adulto , Antiinflamatorios no Esteroideos/efectos adversos , Biomarcadores/sangre , Sedimentación Sanguínea , Proteína C-Reactiva/metabolismo , Estudios de Casos y Controles , Terapia Combinada , Femenino , Humanos , Mediadores de Inflamación/sangre , Masculino , Moxibustión/efectos adversos , Distribución Aleatoria , Espondilitis Anquilosante/sangre , Espondilitis Anquilosante/diagnóstico , Sulfasalazina/efectos adversos , Factores de Tiempo , Resultado del TratamientoRESUMEN
Hepatocellular carcinoma (HCC) is one of the most lethal cancers globally. Hepatitis B virus (HBV) infection might cause chronic hepatitis and cirrhosis, leading to HCC. To screen prognostic genes and therapeutic targets for HCC by bioinformatics analysis and determine the mechanisms underlying HBV-related HCC, three high-throughput RNA-seq based raw datasets, namely GSE25599, GSE77509, and GSE94660, were obtained from the Gene Expression Omnibus database, and one RNA-seq raw dataset was acquired from The Cancer Genome Atlas (TCGA). Overall, 103 genes were up-regulated and 127 were down-regulated. A protein-protein interaction (PPI) network was established using Cytoscape software, and 12 pivotal genes were selected as hub genes. The 230 differentially expressed genes and 12 hub genes were subjected to functional and pathway enrichment analyses, and the results suggested that cell cycle, nuclear division, mitotic nuclear division, oocyte meiosis, retinol metabolism, and p53 signaling-related pathways play important roles in HBV-related HCC progression. Further, among the 12 hub genes, kinesin family member 11 (KIF11), TPX2 microtubule nucleation factor (TPX2), kinesin family member 20A (KIF20A), and cyclin B2 (CCNB2) were identified as independent prognostic genes by survival analysis and univariate and multivariate Cox regression analysis. These four genes showed higher expression levels in HCC than in normal tissue samples, as identified upon analyses with Oncomine. In addition, in comparison with normal tissues, the expression levels of KIF11, TPX2, KIF20A, and CCNB2 were higher in HBV-related HCC than in HCV-related HCC tissues. In conclusion, our results suggest that KIF11, TPX2, KIF20A, and CCNB2 might be involved in the carcinogenesis and development of HBV-related HCC. They can thus be used as independent prognostic genes and novel biomarkers for the diagnosis of HBV-related HCC and development of pertinent therapeutic strategies.
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BACKGROUND: To evaluate the perioperative and long-term results of intrahepatic bile duct exploration lithotomy (IHBDIL) combined with hepatectomy for patients with complicated bilateral primary hepatolithiasis. METHODS: A study was conducted involving 56 patients with complicated bilateral primary hepatolithiasis who underwent IHBDIL combined with hepatectomy at our hospital from January 2006 to December 2014. The perioperative and long-term outcomes that were retrospectively analysed included the stone clearance rate, operative morbidity and mortality, and stone recurrence rate. Patients with a preoperative diagnosis of cholangiocarcinoma were excluded. RESULTS: In all 56 patients, hepatic duct stones were located in the bilateral IHBD. The surgical method was IHBDIL combined with hepatectomy. Postoperative complications occurred in 15 patients (26.8%), 14 patients responded to conservative management, and there was 1 case of postoperative mortality because of hepatic failure. The overall initial success rate of stone clearance was 85.7%, and the final clearance rate was 92.9% following postoperative choledochoscopic lithotripsy. The stone recurrence rate was 13.5%, and the occurrence of postoperative cholangitis was 10.9% during the follow-up period. CONCLUSION: IHBDIL combined with hepatectomy is a safe, effective, and promising treatment for patients with complicated bilateral primary hepatolithiasis. The perioperative and long-term outcomes are satisfactory for complicated bilateral primary hepatolithiasis.
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Conductos Biliares Intrahepáticos/cirugía , Hepatectomía/métodos , Litiasis/cirugía , Hepatopatías/cirugía , Adulto , Anciano , Procedimientos Quirúrgicos del Sistema Biliar , Femenino , Humanos , Laparoscopía/métodos , Hepatopatías/diagnóstico , Masculino , Persona de Mediana Edad , Estudios Retrospectivos , Resultado del TratamientoRESUMEN
To investigate the clinical significance of hepatic parenchyma incision by lithotomy near the second hepatic portal area for the treatment of complex hepatolithiasis.A retrospective study was conducted with 35 patients who had complicated hepatolithiasis in our hospital from January 2008 to December 2013, who underwent hepatic parenchyma incision by lithotomy near the second hepatic portal area. The perioperative and long-term outcomes included the stone clearance rate, operative morbidity and mortality, and the stone recurrence rate. Patients with a preoperative diagnosis of cholangiocarcinoma were excluded from the study.All patients with hepatic duct stones were mainly located at S2, S4, and S8 regions. Surgical methods included were hepatic parenchyma incision by lithotomy near the second hepatic portal area, or by combined partial hepatectomy. The mean follow-up period was 51 months. One patient died during hospitalization. The surgical morbidity was 17.6%, stone clearance rate was 88.2%, and final clearance rate was 94.1% followed by postoperative choledochoscopic lithotripsy. The stone recurrence rate was 15.6% and the occurrence of postoperative cholangitis was 11.8% during the follow-up period.Hepatic parenchyma incision by lithotomy near the second hepatic portal area is safe with satisfactory short and long-term outcome results for complicated hepatolithiasis.
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Procedimientos Quirúrgicos del Sistema Digestivo/métodos , Litiasis/cirugía , Hepatopatías/cirugía , Adulto , Anciano , Femenino , Humanos , Masculino , Persona de Mediana Edad , Mejoramiento de la Calidad , Estudios Retrospectivos , Resultado del TratamientoRESUMEN
OBJECTIVE: To improve Luo-Ye pump-based stress-forming system and optimize the stimulating effect on smooth muscle cells during cultivation of tissue-engineered blood vessels (TEBV). METHODS: A new Luo-Ye pump-based TEBV 3D culture system was developed by adding an air pump to the output of the bioreactor. A pressure guide wire was used to measure the stress at different points of the silicone tube inside the TEBV bio-reactor, and fitting curves of the stress changes over time was created using Origin 8.0 software. The TEBVs were constructed by seeding vascular smooth muscle cells (VSMCs) isolated from human umbilical artery on polyglycolic acid (PGA) and cultured under dynamic conditions with 40 mmHg resistance (improved group), dynamic conditions without resistance (control group) or static condition (static group) for 4 weeks. The harvested TEBVs were then examined with HE staining, masson staining, α-SMA immunohistochemical staining, and scanning and transmission electron microscopy with semi-quantitative analysis of collagen content and α-SMA expression. RESULTS: The measured stress values and the fitting curves showed that the stress stimuli from the Luo-Ye pump were enhanced by adding an air pump to the output of the bioreactor. Histological analysis revealed improved VSMC density, collagen content and α-SMA expression in the TEBVs constructed with the improved method as compared with those in the control and static groups. CONCLUSION: Adding an air pump to the Luo-Ye pump significantly enhances the stress stimulation in the TEBV 3-D culture system to promote the secretion function of VSMCs.
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Reactores Biológicos , Prótesis Vascular , Miocitos del Músculo Liso/citología , Ingeniería de Tejidos/métodos , Células Cultivadas , Colágeno/metabolismo , Humanos , Ácido PoliglicólicoRESUMEN
The circulating tumor cells (CTCs) are derived from primary or metastatic tumor lesions and can be detected in the peripheral blood. With certain specific features, CTCs can,to certain extent, reflect the progression and invasiveness of tumors. Detection of CTCs may provide a powerful and noninvasive approach for diagnosing neoplastic disease, identifying drug sensitivity, and enabling real-time treatment monitoring and prognosis prediction. Improvements in cell isolation and molecular identification will enable a broad range of clinical applications.
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Células Neoplásicas Circulantes , Separación Celular , Progresión de la Enfermedad , Humanos , PronósticoRESUMEN
Long non-coding RNAs(LncRNA)may play a key role in tumorigenesis by regulating gene expression and intervening transcription. Recent studies have demonstrated that a series of patterns including protein modification,chromosomal reconstruction,regulation of target gene expression,transcription intervention,epigenetic modification,and natural antisense transcript are involved in this process. This article reviews recent research advances in this aspect with an attempt to better understand the role of LncRNA in tumorigenesis.
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Transformación Celular Neoplásica , Epigénesis Genética , Regulación Neoplásica de la Expresión Génica , Humanos , ARN Largo no CodificanteRESUMEN
OBJECTIVE: To investigate the mechanism of Advanced glycation end products (AGEs) promoting the calcification of smooth muscle cells. METHODS: The successfully cultured smooth muscle cells were divided into three groups: normal culture group (group A), calcified culture group (group B), calcification + AGEs group (group C); the concentration of intracellular calcium ion was detected in each group; the promotion of AGEs on the calcification of HSMCs was confirmed by VON KOSSA staining; and the expressions of ß-catenin, RAGE, ß-catenin, OPG and E-cadherin protein were detected by immunofluorescence and western blot. RESULTS: The morphology of the cells in each group showed that the amount of calcified plaques in calcification + AGES group were significantly higher than the calcification group. VON KOSSA staining showed that with increasing concentrations of AGE-BSA, the amount of its calcification gradually increased. Calcium concentration in Calcification + 20 mg/L AGEs group was significantly higher, followed by 40 mg/L AGEs group. The expression of ß-catenin increased with the increasing concentrations of AGEs. CONCLUSION: AGEs can promote the calcification of human femoral artery smooth muscle cells, with a concentration gradient effect. With increasing concentrations of AGEs, the expression of RAGE increased, indicating that AGEs-induced HSMCs proliferation was correlated with RAGE expression.
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Cadherinas/metabolismo , Calcinosis , Calcio/metabolismo , Músculo Liso Vascular/patología , beta Catenina/metabolismo , Células Cultivadas , Productos Finales de Glicación Avanzada/metabolismo , Humanos , Músculo Liso Vascular/metabolismo , Miocitos del Músculo Liso/metabolismo , Miocitos del Músculo Liso/patología , Distribución Aleatoria , Albúmina Sérica Bovina/metabolismo , Transducción de SeñalRESUMEN
AIM: To analyze the expression profiles of long non-coding RNAs (lncRNAs) in hepatocellular carcinoma. METHODS: Hepatocellular carcinoma (HCC) tissues and matched adjacent non-tumor (NT) liver tissues were collected from 29 patients with HCC, immediately after liver resection, between March 2011 and July 2013. The diagnosis of HCC was made based on histological examination. Differentially expressed lncRNAs between HCC and NT tissues were revealed through microarray-based lncRNAs expression profiling. Further, quantification of selected lncRNAs was performed using quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR). RESULTS: Six hundred and fifty-nine lncRNAs were differentially expressed between HCC and NT tissues, of which five [TCONS_00018278, AK093543, D16366, ENST00000501583, NR_002819 (MALAT1)] were selected for validation. Four of them were significantly downregulated in HCC tissues compared with NT tissues (P = 0.012, 0.045, 0.000 and 0.000, respectively), and the expression level of MALAT1 showed no significant difference (P = 0.114). CONCLUSION: This study identified a set of lncRNAs differentially expressed in HCC tissues and provided useful information for exploring potential therapeutic targets and diagnostic biomarkers of this cancer.
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Carcinoma Hepatocelular/genética , Perfilación de la Expresión Génica , Regulación Neoplásica de la Expresión Génica , Neoplasias Hepáticas/genética , Análisis de Secuencia por Matrices de Oligonucleótidos/métodos , ARN Largo no Codificante/genética , Adulto , Anciano , Femenino , Humanos , Masculino , Persona de Mediana Edad , Estudios Prospectivos , Reacción en Cadena en Tiempo Real de la Polimerasa , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
Regulations of intracellular protein kinase C (PKC) on carbachol (CCh)-induced intracellular calcium ([Ca(2+)]i) responses were investigated in different stages of melanoma cells. We found that CCh (1 mM) significantly increased [Ca(2+)]i with 6-, 4-, 4-, and 25-folds intensities in WM793B, 451Lu, SK-MEL-5, and A2058 melanoma cells, respectively. Pretreatment of phorbol 12, 13-dibutyrate (PDBu, 2 microM), an activator of intracellular PKC, significantly suppressed CCh-induced peak reactions in WM793B, SK-MEL-5, and A2058 cells. RT-PCR data showed that mRNA levels of PKCalpha were 12-, 4-, 6-, and 0.9-folds higher in above four melanoma cells. Short interfering RNA (siRNA) targeting to PKCalpha in WM793B cells enhanced CCh-induced peak calcium reactions. Present data indicated that CCh-induced [Ca(2+)]i responses were dynamically changed in different stages of melanoma progression. Moreover, intracellular PKCalpha activated by exogenous agonist and expressed through endogenous gene transcription negatively regulated CCh-induced calcium responses. The functional analysis on the relationship between CCh-induced calcium response and endogenous PKCalpha expression might be helpful to predict the development of melanoma.
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Señalización del Calcio/efectos de los fármacos , Carbacol/farmacología , Espacio Intracelular/efectos de los fármacos , Espacio Intracelular/metabolismo , Melanoma/enzimología , Proteína Quinasa C-alfa/metabolismo , Adolescente , Línea Celular Tumoral , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Técnicas de Silenciamiento del Gen , Humanos , Masculino , Melanocitos/efectos de los fármacos , Melanocitos/enzimología , Melanoma/genética , Melanoma/patología , Forbol 12,13-Dibutirato/farmacología , Proteína Quinasa C-alfa/genética , ARN Mensajero/genética , ARN Mensajero/metabolismo , ARN Interferente Pequeño/metabolismo , Transfección , Cicatrización de Heridas/efectos de los fármacosRESUMEN
AIM: To construct the fusion gene of transmembrane programmed death ligand (PD-L1) and red fluorescent protein (DsRed2), and to study the expression and effect of the fused PD-L1/pDsRed2-N1. METHODS: The cloning technology was employed to construct the PD-L1/pDsRed2-N1 fusion gene. The fusion gene was transfected into NIT cells by lipofectamine. The expression of the fusion gene was determined by flow cytometry (FCM) and inverted phase contrast microscope. The effects on allo-spleen cells proliferation were detected by single mixed lymphocyte reaction. FCM was used to detect the proliferation of spleen cells. RESULTS: The expression of the fusion gene was observed in transiently transfected NIT cells. The fusion protein was expressed on cell membrane. These data suggested that the red fluorescent protein tag did not interfere with the natural assembly and the biological activity of PD-L1 molecule. CONCLUSION: The PD-L1-RFP fusion gene is constructed successfully and the fusion protein has biological activity.
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Antígenos de Superficie/metabolismo , Proteínas Reguladoras de la Apoptosis/metabolismo , Proteínas Luminiscentes/metabolismo , Proteínas Recombinantes de Fusión/metabolismo , Animales , Antígenos de Superficie/genética , Proteínas Reguladoras de la Apoptosis/genética , Línea Celular , Membrana Celular/metabolismo , Femenino , Citometría de Flujo , Células Secretoras de Insulina/metabolismo , Proteínas Luminiscentes/genética , Ratones , Ratones Endogámicos BALB C , Microscopía de Contraste de Fase , Embarazo , Receptor de Muerte Celular Programada 1 , Proteínas Recombinantes de Fusión/genética , Proteína Fluorescente RojaRESUMEN
We herein report 2 cases of multiple lobular capillary hemangiomas after scalding. The patients exhibited papules and nodules on the scalded areas after healing. Histopathological examination of the lesions showed capillary proliferation in the upper dermis with edematous stroma containing inflammatory infiltrates predominantly composed of neutrophils. Biopsy tissue and secretion specimens from lesions of case 1 were cultured for bacteria, and both grew Enterobacter cloacae. Ultrastructural examination revealed features typical of a lobular capillary hemangioma and viral inclusion bodies in the epidermis of case 1. Multiple lobular capillary hemangiomas after scalding are rarely reported. Trauma may play an important role in the development of this rare condition. Accumulation of similar cases and its precise observation is needed to confirm the associations and to establish an etiological link between the disease and the pathogens.
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Quemaduras/complicaciones , Granuloma Piogénico/patología , Granuloma Piogénico/fisiopatología , Adulto , Humanos , MasculinoAsunto(s)
Factor de Unión 1 al Potenciador Linfoide/biosíntesis , Melanoma/metabolismo , Neoplasias Cutáneas/metabolismo , Femenino , Humanos , Factor de Unión 1 al Potenciador Linfoide/genética , Masculino , Persona de Mediana Edad , ARN Mensajero/biosíntesis , ARN Mensajero/genética , Transducción de Señal , Proteínas Wnt/genética , Proteínas Wnt/metabolismoRESUMEN
To protect the hematopoietic stem cells (HSCs) from apoptosis induced by chemotherapy and promote HSC proliferation, bi-functional gene delivery systems are increasingly investigated in gene therapy. In the present study, we constructed a bicistronic vector, pWISG, expressing the anti-apoptotic protein human WEE1 (WEE1Hu) and the fusion protein of the proliferation-stimulating stem cell factor (SCF) and enhanced green fluorescent protein (EGFP) separately with internal ribosome entry site (IRES). We first examined the expression and location of WEE1Hu in Chinese hamster ovary (CHO) cells and showed that WEE1Hu was located in the nucleus, which was confirmed by immunohistochemistry and Western blot. We determined the expression and receptor-binding ability of the SCF-EGFP fusion protein on CD34+ cells, which were proved by reverse transcription polymerase chain reaction (RT-PCR) and flow cytometry, respectively. Furthermore, inhibition of cisplatin-induced apoptosis was observed in CD34+ cells transfected with pWISG, which implies that protection for CD34+ cells was achieved via WEE1Hu and SCF-EGFP. Our study suggests that the introduction of two functional genes via bicistronic vector is more powerful and efficient than single gene therapy.