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We aimed to investigate the antitumor effect and mechanism of fructose 1,6-bisphosphate (F1,6BP) in a hepatocellular carcinoma cell line. HepG2 cells were treated with different concentrations of F1,6BP alone or in combination with antioxidant N-acetyl-L-cysteine (NAC) or catalase (CAT), and cell proliferation assays were performed. Nuclear morphology was observed by fluorescence microscopy after Hoechst staining, and apoptosis was measured with flow cytometry. Changes in reactive oxygen species (ROS) levels in HepG2 cells were detected by 2',7'-dichlorodihydrofluorescein diacetate (DCFH-DA) staining. A colorimetric assay was adopted to determine the percentage of oxidized glutathione in these cells. CAT and glutathione peroxidase (GSH-Px) mRNA expression levels in HepG2 cells were measured by real-time fluorescence quantitative PCR. HepG2 cell proliferation was significantly inhibited by F1,6BP, accompanied by an increase in intracellular ROS levels and oxidized glutathione. Upregulated apoptosis and characteristic nuclear morphological changes were observed, and the expression of CAT and GSH-Px mRNA was increased after F1,6BP treatment. The antitumor effect of F1,6BP was significantly decreased after pretreatment with NAC and CAT in HepG2 cells. In conclusion, F1,6BP can induce the apoptosis of HepG2 cells. The mechanism involved may be associated with the generation of ROS, especially the production of H2O2.
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Apoptosis/efectos de los fármacos , Carcinoma Hepatocelular/tratamiento farmacológico , Fructosadifosfatos/farmacología , Neoplasias Hepáticas/tratamiento farmacológico , Acetilcisteína/farmacología , Carcinoma Hepatocelular/genética , Carcinoma Hepatocelular/patología , Catalasa/metabolismo , Proliferación Celular , Regulación Neoplásica de la Expresión Génica , Células Hep G2 , Humanos , Peróxido de Hidrógeno/metabolismo , Especies Reactivas de Oxígeno/metabolismoRESUMEN
Prostate-specific antigen (PSA) serves as a critical biomarker for the early detection and continuous monitoring of prostate cancer. However, commercial PSA detection methods primarily rely on antigen-antibody interactions, leading to issues such as high costs, stringent storage requirements, and potential cross-reactivity due to PSA variant sequence homology. This study is dedicated to the precise design and synthesis of molecular entities tailored for binding with PSA. By employing a million-level virtual screening to obtain potential PSA compounds and effectively guiding the synthesis using machine learning methods, the resulting lead compounds exhibit significantly improved binding affinity compared to those developed before by researchers using high-throughput screening for PSA, substantially reducing screening and development costs. Unlike antibody detection, the design of these small molecules offers promising avenues for advancing prostate cancer diagnostics. Furthermore, this study establishes a systematic framework for the rapid development of customized ligands that precisely target specific protein entities.
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OBJECTIVE: To study the application of umbilical venous catheter (UVC) combined with peripherally inserted central catheter (PICC) in very low birth weight infants (VLBWIs). METHODS: A retrospective analysis was performed on the VLBWIs in the neonatal intensive care unit who received UVC combined with PICC (catheter group, n=63) or did not receive the catheter treatment (non-catheter group, n=38) to compare the differences in nosocomial infection, weight gain, and length of hospital stay between the two groups. RESULTS: The rate of nosocomial infection was 17% in the catheter group and 24% in the non-catheter group (P>0.05). Compared with the non-catheter group, the catheter group had a significantly higher weight gain (11.7±2.0 g/kgâ¢d vs 10.6±2.3 g/kgâ¢d; P<0.05) and a significantly shorter length of hospital stay (40±11 days vs 45±14 days; P<0.05). There was no significant difference in the incidence of complications between the two groups. CONCLUSIONS: Compared with those not receiving catheter treatment, the VLBWIs receiving UVC combined with PICC have a markedly higher weight gain and a markedly shorter length of hospital stay and show a declining trend in the rate of nosocomial infection.
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Cateterismo Periférico , Catéteres Venosos Centrales , Recién Nacido de muy Bajo Peso , Cateterismo Periférico/efectos adversos , Infección Hospitalaria/epidemiología , Humanos , Recién Nacido , Estudios Retrospectivos , Venas UmbilicalesRESUMEN
OBJECTIVE: To investigate the high-risk factors for parenteral nutrition-associated cholestasis (PNAC), which is the most common complication of parenteral nutrition for infants, in very low birth weight infants (VLBWIs). METHODS: Retrospective analysis was performed on the clinical and laboratory data of 204 VLBWIs who received parenteral nutrition for over 2 weeks in the neonatal intensive care unit from August 2006 to December 2011. The infants'liver function was evaluated periodically before and after Parenteral nutrition. Univariate analysis and multivariate analysis were performed in the observation (PNAC) and control (without PNAC) groups. RESULTS: PNAC occurred in 46 (22.5%) of the 204 VLBWIs. Univariate analysis showed that continuous positive airway pressure (CPAP) ventilation, respiration failure, respiratory distress syndrome (RDS), bronchopulmonary dysplasia (BPD) and retinopathy of prematurity (ROP) were significantly increased in the observation group compared with the control group. The observation group had lower birth weights, longer duration of ventilation, later breast feeding beginning, longer duration of fasting, longer duration of parenteral nutrition, and higher cumulated amino acid and lipid emulsion intake. Logistic regression analysis revealed that duration of fasting was a high-risk factor for PNAC (OR=1.115, 95%CI: 1.031-1.207). CONCLUSIONS: Many risk factors are associated with PNAC. Early enteral nutrition and short duration of parenteral nutrition are helpful in preventing the incidence of PNAC in VLBWIs.
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Colestasis/etiología , Recién Nacido de muy Bajo Peso , Nutrición Parenteral/efectos adversos , Colestasis/prevención & control , Humanos , Recién Nacido , Modelos Logísticos , Factores de RiesgoRESUMEN
OBJECTIVES: Endoscopic retrograde cholangiopancreatography (ERCP) in patients with Roux-en-Y reconstruction after total gastrectomy is difficult to be performed using routine tools. The aim of this study was to evaluate the feasibility and safety of cap-assisted routine adult colonoscope (CARAC) for ERCP in these patients. METHODS: Sixteen consecutive patients with indications of ERCP who had previously undergone total gastrectomy with Roux-en-Y reconstruction at two tertiary care endoscopy centers were identified. All ERCP procedures were carried out by using CARAC. The success rate of reaching the papilla, biliary cannulation and procedure-related adverse events were analyzed. RESULTS: The papilla was successful reached in 11 (68.8%) of the 16 cases, and biliary cannulation was subsequently reached in eight (72.7%) of the 11 cases. The procedures succeeded in three patients by using a percutaneous-endoscopic rendezvous procedure after failed cannulation. Overall clinical success was achieved in 11 (68.8%) of 16 patients. Procedure-related mild acute pancreatitis was observed in 25.0% (4/16) of the cases and mild cholangitis in 18.8% (3/16). No serious adverse events were reported. CONCLUSIONS: CARAC for therapeutic ERCP is safe and effective in treating patients with Roux-en-Y reconstruction after total gastrectomy.
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Colangiopancreatografia Retrógrada Endoscópica , Pancreatitis , Enfermedad Aguda , Colonoscopios , Estudios de Factibilidad , Gastrectomía/efectos adversos , HumanosRESUMEN
BACKGROUND: Dihydroartemisinin (DHA), a semisynthetic derivative of artemisinin, has a broad range of biological properties, including antitumor activity. However, the mechanisms by which DHA affects the tumorigenesis of gastric carcinoma (GC) are poorly understood. MATERIAL AND METHODS: The targets of DHA were identified by network pharmacology, and the association of CDK4 with clinicopathological characteristics and prognosis in patients with GC was analyzed by using TCGA data. CCK8, Transwell and flow cytometric analyses, as well as a tumor xenograft model, were used to assess the effects of DHA on the growth and migration of GC cells. qRT-PCR and Western blot analyses were used to determine the effects of DHA on the cyclin D1-CDK4-Rb signaling pathway. RESULTS: We identified 13 DHA targets and measured their expression of whichCDK4 expression levels were substantially higher in GC tissues than those in adjacent normal tissues, and high CDK4 expression acted as an independent prognostic factor of poor survival in patients with GC. DHA suppressed cell proliferation, migration and invasion in vitro and in vivo and induced G1 phase cell cycle arrest in a dose-dependent manner by regulating cyclin D1-CDK4-Rb signaling. CONCLUSIONS: DHA inhibits the tumorigenesis and invasion of GC by regulating cyclin D1-CDK4-Rb signaling and may provide therapeutic strategies for the treatment of GC.
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Artemisininas/farmacología , Ciclina D1/metabolismo , Quinasa 4 Dependiente de la Ciclina/metabolismo , Transducción de Señal/efectos de los fármacos , Neoplasias Gástricas/tratamiento farmacológico , Animales , Artemisininas/química , Carcinogénesis/efectos de los fármacos , Puntos de Control del Ciclo Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Ciclina D1/genética , Quinasa 4 Dependiente de la Ciclina/genética , Femenino , Humanos , Masculino , Ratones , Ratones Endogámicos BALB C , Ratones Desnudos , Persona de Mediana Edad , Invasividad Neoplásica/prevención & control , Pronóstico , Estómago/patología , Neoplasias Gástricas/diagnóstico , Neoplasias Gástricas/patología , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
Studies in our laboratory have shown that a fraction of apolipoprotein (apo) E internalized by hepatocytes escapes degradation and is resecreted. Although the intracellular routing is not fully understood, our studies suggest that a portion of apoE recycles through the Golgi apparatus. Given the role of the Golgi apparatus in lipoprotein secretion and the fact that apoE modulates the hepatic secretion of very low-density lipoprotein, we hypothesized that recycling apoE has an effect on hepatic very low-density lipoprotein assembly and/or secretion. To test this hypothesis, apoE-/- mice were transplanted with bone marrow from wild-type mice. In this model, extrahepatic (macrophage-derived) apoE is internalized by the hepatocytes in vivo and is resecreted when the hepatocytes are placed in culture. Unexpectedly, our studies demonstrate that recycling apoE has little effect on hepatic lipid content or hepatocyte triglyceride secretion. In addition, recycling apoE has little effect on the expression of enzymes and proteins involved in lipid synthesis as well as plasma lipoprotein apoproteins. We conclude that the physiological relevance of apoE recycling may not be related to cell-specific functions, such as lipoprotein assembly in the liver. Rather, recycling may provide a mechanism for modulating general cellular effects such as intracellular cholesterol transport or cholesterol efflux.
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Apolipoproteínas E/metabolismo , Hepatocitos/metabolismo , Animales , Trasplante de Médula Ósea , Proteínas Potenciadoras de Unión a CCAAT/genética , Células Cultivadas , Proteínas de Unión al ADN/genética , Ácidos Grasos/análisis , Lipoproteínas VLDL/metabolismo , Ratones , Ratones Endogámicos C57BL , ARN Mensajero/análisis , Proteína 1 de Unión a los Elementos Reguladores de Esteroles , Factores de Transcripción/genética , Triglicéridos/análisisRESUMEN
A murine epididymal retinoic-acid-binding protein (mE-RABP) is specifically expressed in the mid/distal caput epididymidis and is androgen regulated. The murine epididymal protein of 17 kDa (mEP17) gene, a novel gene homologous to mE-RABP, is located within 5 kb of the 5'-flanking region of the mE-RABP gene. In contrast, expression of the mEP17 gene is restricted to the initial segment and regulated by factor(s) contained in testicular fluid. To identify cis-DNA regulatory element(s) involved in the tissue- and region-specific expression of the mEP17 gene in transgenic mice, we have studied the expression of a transgene containing 5.3 kb of the 5'-flanking region of the mEP17 gene (5.3mEP17) linked to chloramphenicol acetyltransferase (CAT) reporter gene. Significant caput epididymidis-specific CAT activity was detected in transgenic mouse lines; and CAT gene expression is restricted to the initial segment, as is the expression of the endogenous mEP17 gene. Ontogenic expression and testicular factor dependency also mimic that of endogenous mEP17 gene. These results suggest that the 5.3mEP17 fragment contains all the information required for spatial and temporal expression in the mouse epididymis. The 5.3mEP17 fragment will be useful to express a foreign gene of interest in the epididymis in an initial segment-specific manner.
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Proteínas Portadoras/genética , Epidídimo/química , Epidídimo/metabolismo , Expresión Génica , Transgenes/genética , Animales , Cloranfenicol O-Acetiltransferasa/genética , Genes Reporteros , Hibridación in Situ , Lipocalinas , Masculino , Ratones , Ratones Transgénicos , Datos de Secuencia Molecular , Fragmentos de Péptidos/genética , ARN Mensajero/análisis , Receptores de Ácido Retinoico/genética , Proteínas Recombinantes de Fusión , Secuencias Reguladoras de Ácidos Nucleicos , Mapeo RestrictivoRESUMEN
OBJECTIVE: To investigate the dynamic progress of 2,4,6-trinitrobenzene sulfonic acid (TNBS)-induced chronic colitis and fibrosis in rat model. METHODS: In all, 44 Sprague-Dawley rats were randomly divided into the model and control groups. Colitis was induced by intrarectal injection of 10-30 mg TNBS in 50% ethanol enema weekly for 5 cycles. The control group received an equal volume of 50% ethanol. If the rat died during the procedure, necropsy was performed immediately. At the end of the 2nd, 3rd, 4th and 5th week the rats were sacrificed, and histological damage and fibrosis of the colon were examined using HE and Masson trichrome stain. The concentrations of Th1, Th2, Th17 cytokines in colon tissue were detected by ELISA, intestinal fibrosis-relevant cytokine expressions were detected by fluorescent quantification-polymerase chain reaction. RESULTS: Colitis model was successfully induced with a low mortality rate. The microscopic colonic damage score, collagen area, Th1/Th17 cytokines and expressions of intestinal fibrosis-relevant cytokines were significantly higher in the model group than those in the control group. Furthermore, the collagen area, content of interleukin 17 and expressions of intestinal fibrosis-related cytokines in the model group were more elevated in the chronic phase (after 3 to 4 cycles) than in the acute phase (P < 0.05). CONCLUSIONS: Multiple inflammatory responses participate in the formation and dynamic progression of TNBS-induced chronic colitis. In particular, acute colitis may turn into chronic colitis after 3 cycles of TNBS administration. This coincides with the formation of intestinal fibrosis which is concomitantly exacerbated after cycle 4.
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Colitis/metabolismo , Colitis/patología , Colon/metabolismo , Colon/patología , Citocinas/metabolismo , Análisis de Varianza , Animales , Enfermedad Crónica , Colitis/inducido químicamente , Colágeno Tipo I/genética , Colágeno Tipo I/metabolismo , Colágeno Tipo III/genética , Colágeno Tipo III/metabolismo , Factor de Crecimiento del Tejido Conjuntivo/genética , Factor de Crecimiento del Tejido Conjuntivo/metabolismo , Citocinas/genética , Femenino , Fibrosis , Expresión Génica , Interleucina-13/genética , Interleucina-13/metabolismo , Interleucina-17/metabolismo , Interleucina-2/metabolismo , Interleucina-4/metabolismo , ARN Mensajero/metabolismo , Ratas , Ratas Sprague-Dawley , Proteína smad3/genética , Proteína smad3/metabolismo , Factores de Tiempo , Factor de Crecimiento Transformador beta1/genética , Factor de Crecimiento Transformador beta1/metabolismo , Ácido Trinitrobencenosulfónico , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
OBJECTIVE: To explore the application of repeated measurement 'analysis of variance' (ANOVA) in evaluating the effectiveness of 'community-based hypertension self-management program'. METHODS: A community-based parallel controlled trial was conducted among 3 communities. 169 patients in intervention group took part in the course on hypertension self-management program once a week and 204 patients in control group received routine hypertension management services. Data collected through questionnaire at baseline and 6 months, 12 months after intervention and were compared through repeated ANOVA measurement. RESULTS: Subjects in the intervention group showed statistical significance and linear trends in health self-evaluation, distress, in low spirit, self-efficacy in managing symptoms (SEMS), self-efficacy to managing diseases in general (SEMDG), systolic blood pressure (SBP) and diastolic blood pressure (DBP) over time by univariate test of repeated measurement ANOVA. The score of SEMS increased from 6.84 +/- 2.53 at baseline to 8.20 +/- 1.44 at 12 months after intervention while SEMDG from 7.28 +/- 2.45 to 8.89 +/- 1.05, and SBP decreased from 137.66 +/- 7.30 mm Hg (1 mm Hg = 0.133 kPa) to 130.41 +/- 7.71 mm Hg, DBP decreased from 84.13 +/- 6.70 mm Hg to 81.04 +/- 5.98 mm Hg respectively. Only low spirit and SBP changed over time were seen in the control group. Self-evaluation, distress,in low spirit, caused by diseases, SEMS, SEMDG and SBP were statistically different between control and intervention groups, and the effect of interaction between groups and time span were statistically significant on indicators as self-evaluation, low spirit, self-efficacy in managing symptoms, self-efficacy to manage diseases and SBP etc, by multivariate test of repeated measurement ANOVA. CONCLUSION: Repeated measurement ANOVA not only could be used to analyze group-effect, but could also explain the effect and the interaction among groups and time, to make the results more reliable. The self-management approach could improve the health status and self-efficacy of the patients,so as to reduce the blood pressure. Our result showed that it was effective for hypertensive patients to be on the chronic diseases self-management program.
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Hipertensión/prevención & control , Autocuidado , Anciano , Análisis de Varianza , Femenino , Humanos , Masculino , Persona de Mediana Edad , Resultado del TratamientoRESUMEN
Microsomal triglyceride transfer protein (MTP) has been studied extensively, primarily because of its role in the assembly of very low density lipoproteins by the liver and chylomicrons by the intestine. Recent studies have suggested that MTP may also play key roles in other cellular processes. In this paper we report the identification of a novel splice variant of MTP in mice. This isoform, MTP-B, has a unique first exon located approximately 2.7 kilobases upstream of canonical MTP (MTP-A) exon 1. The alternative exon encodes 35 amino acids compared with 20 amino acids encoded by exon 1 of MTP-A. MTP-B represents approximately 90% of total MTP mRNA in mouse adipocytes and 3T3-L1 cells and <5% in mouse liver and intestine. Expression of the alternate isoform in mouse liver was confirmed by mass spectrometry. Co-transfection of COS cells with truncated forms of apoB and either MTP-A or MTP-B demonstrated that both isoforms are effective in the assembly and secretion of nascent apoB-containing lipoproteins. Confocal microscopy of 3T3-L1 cells transfected with enhanced green fluorescent protein or DsRed fusions of the two proteins revealed that MTP-A is localized to the endoplasmic reticulum, whereas MTP-B localizes primarily to the Golgi complex in these cells. We conclude that MTP-B functions similarly to MTP-A in lipoprotein assembly. However, in nonlipoprotein-secreting cells, such as the adipocyte, MTP-B may have different localization properties, perhaps reflecting a distinct role in lipid storage and mobilization.
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Proteínas Portadoras/aislamiento & purificación , Células 3T3-L1 , Secuencia de Aminoácidos , Animales , Células COS , Proteínas Portadoras/química , Proteínas Portadoras/genética , Proteínas Portadoras/fisiología , Chlorocebus aethiops , Espectrometría de Masas , Ratones , Ratones Endogámicos C57BL , Datos de Secuencia Molecular , Isoformas de Proteínas , TransfecciónRESUMEN
OBJECTIVE: This study aimed to understand the prevalence rate, epidemiological characteristics and relevant factors of arthritis in Shanghai. METHODS: A sample of 7 575 residents aged 15 years and above was drawn from 6 communities under multiple stage cluster sampling. A household survey with questionnaire was carried out to differentiate both undiagnosed patients and those with definite arthritis. Those who had not been diagnosed before were asked to carry further clinical examinations by a rheumatologist. RESULTS: The prevalence rate of arthritis was 6.11%, including osteoarthritis (OA) 4.18%, rheumatoid arthritis (RA) 0.52%, gout 0.28%, ankylosing spondylitis (AS) 0.28%, rheumatic arthritis 0.49% and other types arthritis 0.82%. Arthritis was significantly related to cardiovascular disease, pulmonary disease and gastrointestinal disease. Age, female and obesity might serve as risk factors for arthritis. Physical labors and living in rural area might have protecting effects. CONCLUSION: Elderly and female seemed to be at high risk for arthritis. Weight control and more exercise should be encouraged to reduce the risks. For arthritis patients, treatment to other chronic diseases should not be ignored.
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Artritis/epidemiología , Adolescente , Adulto , Factores de Edad , Anciano , Anciano de 80 o más Años , Artritis/clasificación , Artritis/complicaciones , China/epidemiología , Femenino , Humanos , Masculino , Persona de Mediana Edad , Prevalencia , Factores de Riesgo , Factores Sexuales , Clase Social , Factores SocioeconómicosRESUMEN
Microsomal triglyceride transfer protein (MTP) is essential for the assembly of apolipoprotein B-containing lipoproteins. Within the endoplasmic reticulum, it transfers lipid from the membrane to the forming lipoprotein. Recent evidence suggests that it may also function within the Golgi apparatus. To address this hypothesis, we developed a polyclonal antibody to MTP and used it in a series of studies on mouse liver and McArdle-RH7777 (McA) cells. Western blot analysis demonstrated the presence of MTP within mouse hepatic-Golgi apparatus-rich fractions. In addition, in vitro lipid transfer assays demonstrated the presence of triglyceride transfer activity within the Golgi fractions. Immunohistochemical studies with mouse liver demonstrated the presence of MTP within all hepatocytes, but not in nonparenchymal cells. The subcellular location of MTP in McA cells was investigated using confocal microscopy. MTP colocalized with the trans-Golgi network (TGN) 38 and Golgi SNARE (soluble N-ethylmalemide-sensitive factor attachment protein receptor) of 28 kDa (GS28), markers for the trans- and cis-Golgi apparatus, respectively. Morphometric analyses indicated that approximately 17% of the MTP signal colocalized with the TGN38, while 33% of the trans-Golgi marker colocalized with the MTP. Approximately 17% of the MTP signal colocalized with the GS28, whereas 53% of the cis-Golgi marker colocalized with the MTP. The results provide unequivocal evidence for the location of MTP within the Golgi apparatus, and further highlight the importance of this organelle in the assembly of lipoproteins.