RESUMEN
BACKGROUND: There is a long latent period for the sciatic nerve block before a satisfactory block is attained. Changes in the temperature of local anesthetics may influence the characters of the peripheral nerve block. This study was designed to evaluate the effect of warming ropivacaine on the ultrasound-guided subgluteal sciatic nerve block. METHODS: Fifty-four patients for distal lower limbs surgery were randomly allocated into warming group (group W, n = 27) or room tempeture group (group R, n = 27) with the ultrasound-guided subgluteal sciatic nerve block. The group W received 30 ml of ropivacaine 0.5% at 30â and the group R received 30 ml of ropivacaine 0.5% at 23â. The sensory and motor blockade were assessed every 2 min for 30 min after injection. The primary outcome was the onset time of limb sensory blockade. RESULTS: The onset time of sensory blockade was shorter in group W than in group R (16 (16,18) min vs 22 (20,23) min, p < 0.001), and the onset time of motor blockade was also shorter in group W than in group R (22 (20,24) min vs 26 (24,28) min, p < 0.001). The onset time of sensory blockade for each nerve was shorter in group W than in group R (p < 0.001). No obvious differences for the duration of sensory and motor blockade and the patient satisfaction were discovered between both groups. No complications associated with nerve block were observed 2 days after surgery. CONCLUSIONS: Warming ropivacaine 0.5% to 30â accelerates the onset time of sensory and motor blockade in the ultrasound-guided subgluteal sciatic nerve block and it has no influence on the duration of sensory and motor blockade. TRIAL REGISTRATION: The trial was registered on October 3, 2022 in the Chinese Clinical Trial Registry ( https://www.chictr.org.cn/bin/project/edit?pid=181104 ), registration number ChiCTR2200064350 (03/10/2022).
Asunto(s)
Amidas , Nervio Ciático , Humanos , Ropivacaína/farmacología , Amidas/farmacología , Nervio Ciático/diagnóstico por imagen , Anestésicos Locales/farmacología , Ultrasonografía IntervencionalRESUMEN
Choroidal neovascularization (CNV) is a major cause of vision loss in many retinal diseases. Hypoxia is determined to be a key inducer of CNV and hypoxia-inducible factor-1 (HIF-1) is an important transcription factor. Epithelial-mesenchymal transition (EMT) and the synthesis of proangiogenic cytokines make great contributions to the development of CNV. In the present study, the role of HIF-1α signaling in the regulation of angiogenin (ANG) expression and EMT in hypoxic retinal pigment epithelial cells was investigated. A significant elevation expression of ANG expression level in a mouse model of laser-induced CNV was demonstrated. In a hypoxic model of ARPE-19, an increased expression level of ANG and induction of EMT accompanied with stabilization and nucleus translocation of HIF-1α. Blockage of HIF-1α signaling resulted in inhibition of high expression of ANG and EMT features. The direct interaction between HIF-1α and ANG promoter region was identified by ChIP-qPCR. The association of RNase 4 mRNA level with HIF-1α signaling was also clarified in APRE-19. Moreover, the exogenous ANG translocated into the nucleus, enhanced 45S rRNA transcription, promoted cell proliferation and tube formation in human retinal microvascular endothelial cells. In conclusion, the hypoxic conditions regulate the expression of ANG and EMT via an activation of HIF-1α signaling. It provides molecular evidence for potential therapy strategies of treating CNV.
Asunto(s)
Transición Epitelial-Mesenquimal/fisiología , Subunidad alfa del Factor 1 Inducible por Hipoxia/metabolismo , Epitelio Pigmentado de la Retina/metabolismo , Epitelio Pigmentado de la Retina/patología , Ribonucleasa Pancreática/genética , Animales , Hipoxia de la Célula/fisiología , Línea Celular , Neovascularización Coroidal/etiología , Neovascularización Coroidal/metabolismo , Neovascularización Coroidal/patología , Modelos Animales de Enfermedad , Regulación de la Expresión Génica , Humanos , Ratones , Ratones Endogámicos C57BL , Modelos Biológicos , Transducción de Señal , Regulación hacia ArribaRESUMEN
BACKGROUND/AIMS: Macrophage inflammatory protein-2 (MIP-2), a type of leukocyte chemokine, is primarily produced by macrophages, and levels increase significantly in early inflammation. However, the precise biological functions and mechanisms of MIP-2 in the development of inflammation remain unclear. The purposes of the present study were to investigate the role of MIP-2 in inflammation induced by lipopolysaccharide (LPS) in vitro and to determine the possibility of blocking the high mobility group box 1 (HMGB1) signalling pathway via MIP-2 inhibition. METHODS: Macrophage cells (RAW264.7, U937 and THP-1 cells) were divided into control and treatments groups. Expression levels of interleukin-6 (IL-6), interleukin-1ß (IL-1ß), tumour necrosis factor-α (TNF-α), HMGB1, chemokine (C-C motif) ligand-2 (Ccl-2), Toll-like receptor-4 (TLR-4), inducible nitric oxide synthase (iNOS), phosphorylated MAPKs (p38, ERKs, JNKs), PI3K/Akts, JAKs/STAT3, IκB, and cytoplasmic and nuclear NF-κB p65 in RAW264.7 cells were detected by qRT-PCR, enzyme-linked immunosorbent assay (ELISA) or western blot assays. RESULTS: mip-2 siRNA and an anti-MIP-2 antibody significantly reduced the expression levels of Ccl-2, TLR-4, iNOS, IL-6, IL-1ß, HMGB1, and TNF-α in RAW264.7 cells exposed to LPS (P<0.01). Additionally, mRNA expression levels of HMGB1 and TLR-4 in cells treated with LPS+mip-2 siRNA were significantly lower than those in cells treated with LPS alone (P<0.01 or P<0.05). The MIP-2 antibody significantly suppressed activation of p38-MAPK, p-STAT3, and p-Akts and translocation of NF-κB p65 from the cytoplasm to the nucleus in RAW264.7 exposed to LPS (P<0.01 or P<0.05). CONCLUSION: mip-2 siRNA and the MIP-2 antibody can reduce the inflammatory effects induced by LPS in macrophage cells. The mechanisms may occur through down-regulation of p38-MAPK, STAT3 and Akts phosphorylation and translocation of NF-κB p65. MIP-2 plays an important role in inflammation induced by LPS.
Asunto(s)
Quimiocina CXCL2/inmunología , Proteína HMGB1/inmunología , Inflamación/inmunología , Lipopolisacáridos/inmunología , Macrófagos/inmunología , Interferencia de ARN , Animales , Quimiocina CXCL2/genética , Regulación hacia Abajo , Regulación de la Expresión Génica , Proteína HMGB1/genética , Inflamación/genética , Interleucina-1beta/genética , Interleucina-6/genética , Macrófagos/metabolismo , Ratones , Óxido Nítrico Sintasa de Tipo II/genética , Células RAW 264.7 , ARN Interferente Pequeño/genética , Receptor Toll-Like 4/genética , Factor de Necrosis Tumoral alfa/genéticaRESUMEN
BACKGROUND The aim of this study was to explore the efficacy of temporary balloon occlusion of the abdominal aorta assisting open reduction and internal fixation (ORIF) in the treatment of complex acetabular fracture. MATERIAL AND METHODS From August 2000 to October 2011, a total of 48 patients with complex acetabular fracture were enrolled in this study. Average operative time, intraoperative blood loss volume, blood transfusion volume, satisfactory reduction, and postoperative functional recovery rate were recorded and compared between the 2 groups. RESULTS A significant difference was observed between the 2 groups in operative time (P=0.003). For intraoperative blood loss and blood transfusion, ORIF combined with temporary balloon occlusion of abdominal aorta techniques appeared to be superior to normal ORIF (blood loss: P=0.007; and blood transfusion: P=0.019, respectively). However, no differences were observed in postoperative blood loss or transfusion (P>0.05). Patients in group A showed better hip function than those in group B (group A: a good-to-excellent rate of 77.8%; group B: a good-to-excellent rate of 78.3%; P>0.05). With regard to the incidence of postoperative complications, there were no significant differences between the 2 groups (group A: 9/18; group B: 11/23; P=0.890). CONCLUSIONS In the treatment of complex acetabular fracture, temporary balloon occlusion of the abdominal aorta is a reliable technique to assist ORIF surgery to staunch the flow of blood.
Asunto(s)
Acetábulo/cirugía , Aorta Abdominal/cirugía , Oclusión con Balón/métodos , Fijación Interna de Fracturas/métodos , Fracturas Óseas/cirugía , Acetábulo/lesiones , Adulto , Pérdida de Sangre Quirúrgica , Femenino , Humanos , Masculino , Complicaciones Posoperatorias/etiología , Estudios Retrospectivos , Resultado del TratamientoRESUMEN
BACKGROUND/AIMS: To investigate the effects of emodin on concanavalin A (Con A)-induced hepatitis in mice and to elucidate its underlying molecular mechanisms. METHODS: A fulminant hepatitis model was established successfully by the intravenous administration of Con A (20 mg/kg) to male Balb/c mice. Emodin was administered to the mice by gavage before and after Con A injection. The levels of pro-inflammatory cytokines and chemokines, numbers of CD4(+) and F4/80(+) cells infiltrated into the liver, and amounts of phosphorylated p38 MAPK and NF-κB in mouse livers and RAW264.7 and EL4 cells were measured. RESULTS: Pretreatment with emodin significantly protected the animals from T cell-mediated hepatitis, as shown by the decreased elevations of serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST), as well as reduced hepatic necrosis. In addition, emodin pretreatment markedly reduced the intrahepatic expression of pro-inflammatory cytokines and chemokines, including tumor necrosis factor (TNF)-α, interferon (IFN)-γ, interleukin (IL)-1ß, IL-6, IL-12, inducible nitric oxide synthase (iNOS), integrin alpha M (ITGAM), chemokine (C-C motif) ligand 2 (CCL2), macrophage inflammatory protein 2 (MIP-2) and chemokine (CXC motif) receptor 2 (CXCR2). Furthermore, emodin pretreatment dramatically suppressed the numbers of CD4(+) and F4/80(+) cells infiltrating into the liver as well as the activation of p38 MAPK and NF-κB in Con A-treated mouse livers and RAW264.7 and EL4 cells. CONCLUSION: The results indicate that emodin pretreatment protects against Con A-induced liver injury in mice; these beneficial effects may occur partially through inhibition of both the infiltration of CD4(+) and F4/80(+) cells and the activation of the p38 MAPK-NF-κB pathway in CD4(+) T cells and macrophages.
Asunto(s)
Emodina/farmacología , Sustancias Protectoras/farmacología , Transducción de Señal/efectos de los fármacos , Alanina Transaminasa/sangre , Animales , Aspartato Aminotransferasas/sangre , Antígeno CD11b/genética , Antígeno CD11b/metabolismo , Linfocitos T CD4-Positivos/citología , Linfocitos T CD4-Positivos/inmunología , Línea Celular , Quimiocinas/genética , Quimiocinas/metabolismo , Concanavalina A/inmunología , Concanavalina A/toxicidad , Citocinas/genética , Citocinas/metabolismo , Hepatitis/etiología , Hepatitis/metabolismo , Hepatitis/patología , Hígado/efectos de los fármacos , Hígado/metabolismo , Masculino , Ratones , Ratones Endogámicos BALB C , FN-kappa B/metabolismo , Óxido Nítrico Sintasa de Tipo II/genética , Óxido Nítrico Sintasa de Tipo II/metabolismo , Fosforilación/efectos de los fármacos , Proteínas Quinasas p38 Activadas por Mitógenos/metabolismoRESUMEN
So far, several studies on the association between osteopontin (OPN) expression and glioma have been performed, but the conclusion still was not clear. The aim of the present meta-analysis was to determine the relationship between OPN expression and prognosis of patients with glioma. The electronic database was searched for articles on the association between OPN expression and glioma until 31 January 2014. Odds ratios (OR) and the relative risks with 95 % CI were utilized to analyze the qualitative data in retrospective studies and prospective studies, respectively. The standardized mean difference and the corresponding 95 % CI were used for analyzing the studies with quantitative data. Heterogeneity of all included studies was assessed using Cochrane's Q test and I (2) measurement. The publication bias was examined by the Egger test. Sixteen cohort studies (854 patients) on OPN expression and gliomas prognosis were included in the present meta-analysis. It was found that OPN expression was significantly higher in patients with high-grade glioma than in patients with low-grade glioma (χ (2) = 8.38, I (2) = 16.6 %, P = 0.300), and the expression of OPN increased with glioma grade. The combined data showed the correlation between high OPN expression and tumor reoccurrence (OR = 18.61, 95 % CI = 6.34-54.67, P = 0.405). In addition, the results of the pooled analysis indicated that OPN expression was significantly related to overall survival (HR = 1.83; 95 % CI = 1.36-2.46). In conclusion, OPN may be a biomarker for predicting the prognosis of gliomas.
Asunto(s)
Biomarcadores de Tumor/metabolismo , Neoplasias Encefálicas/metabolismo , Glioma/metabolismo , Osteopontina/metabolismo , Neoplasias Encefálicas/mortalidad , Neoplasias Encefálicas/patología , Glioma/mortalidad , Glioma/patología , Humanos , Pronóstico , Análisis de Supervivencia , Carga TumoralRESUMEN
BACKGROUND: Acute liver failure (ALF), known as a rapid and severe clinical syndrome, can induce multiple organ dysfunction and failure. It was noticed that Kupffer cells activation at the initial phase was involved in some intense inflammatory responses in the pathogenesis of ALF. However, detailed regulation mechanism of Kupffer cells activation during ALF is still obscured. Present study aimed to discover the potential regulator and explore deeper information of Kupffer cells activation at the early stage of ALF. METHODS: The mouse model of ALF was established by Concanavalin A injection. Dynamic immunological statuses of Kupffer cells at the early stage of ALF were exhibited by detecting typical cytokines. The expression of inflammasome AIM2 was measured in both RNA and protein level. Its role of affecting Kupffer cells activation during ALF by inducing IL-1ß production was identified by RNA interference in vitro. Moreover, the expression of miR-223 in vivo was measured by q-PCR and its role in regulating Kupffer cells activation during Con A induced ALF was determined by RNAs transfection. RESULTS: Present study showed that mass production of IL-1ß from isolated Kupffer cells in Con A treated mice might be the main driving force of Kupffer cells pro-inflammatory activation during ALF. The role of AIM2 in affecting pro-inflammatory activation of Kupffer cells by inducing IL-1ß production was crucial to ALF. Further study found that miR-223 acted as a regulator in Kupffer cells activation at the early stage of ALF by influencing IL-1ß production via AIM2 pathway. CONCLUSION: For the first time, this paper demonstrated that miR-223 acted to inhibit IL-1ß production via AIM2 pathway, suppressing Kupffer cells pro-inflammatory activation at the early stage of ALF. Thus, it played an important role in the pathogenesis of ALF.
Asunto(s)
Proteínas de Unión al ADN/genética , Macrófagos del Hígado/patología , Fallo Hepático Agudo/genética , MicroARNs/genética , Animales , Proteínas de Unión al ADN/biosíntesis , Regulación de la Expresión Génica , Humanos , Interleucina-1beta/genética , Fallo Hepático Agudo/inducido químicamente , Fallo Hepático Agudo/patología , Ratones , MicroARNs/biosíntesis , Transducción de SeñalRESUMEN
BACKGROUND: Resveratrol (Res) is a polyphenol anti-inflammatory agent. We have studied the link between the anti-inflammatory effects of Res and the high mobility group box 1(HMGB1) signaling pathway. METHODS: Murine macrophage-like RAW264.7 cells (RAW264.7 cells) were either untreated (control) or treated with Res, LPS, or LPS + Res. Levels of IL-6, NO, and TNF-α were measured by ELISA and colorimetric assays. Expression of HMGB1 was detected by qRT-PCR, western blot, and immunofluorescence assays. Protein and mRNA expression levels of TLR4 were also examined. RESULTS: Res significantly reduced the levels of IL-6, NO, and TNF-α in RAW264.7 cells exposed to LPS. Expression levels of HMGB1 (mRNA and protein) and of TLR4 in the LPS + Res-treated cells were lower than in cells treated with LPS alone. CONCLUSIONS: Res can block the inflammatory effects induced by LPS in RAW264.7 cells. Down-regulation of HMGB expression may be one of the mechanisms of action of Res. Res may also influence TLR4 expression in the HMGB1-TLR4 signaling pathway.
Asunto(s)
Antiinflamatorios no Esteroideos/farmacología , Proteína HMGB1/genética , Inflamación/tratamiento farmacológico , Lipopolisacáridos/farmacología , Estilbenos/farmacología , Receptor Toll-Like 4/genética , Animales , Línea Celular , Relación Dosis-Respuesta a Droga , Lipopolisacáridos/antagonistas & inhibidores , Ratones , Estructura Molecular , ARN Mensajero/biosíntesis , ARN Mensajero/genética , Resveratrol , Estilbenos/uso terapéutico , Relación Estructura-ActividadRESUMEN
More than a third of the world's population is infected with the hepatitis B virus (HBV) and 5% are thought to be HBV carriers, putting them at risk of developing serious liver diseases. The treatment of liver diseases with Chinese herbal medicines (CHM) dates back 2,500 years and the aim of this analysis was to evaluate the efficacy and safety of CHM for HBV carriers compared to Western medicine (WM) or placebo and to summarize the most commonly used herbs. Several databases, such as Pubmed, Embase and the Chinese database CNKI, were used to evaluate randomized, controlled trials (RCTs) focused on CHM treatment for HBV carriers up to 2013. We performed a systematic review and meta-analysis on the herbs and their effect on hepatitis B viral proteins (HBeAg, HBsAg) and HBV DNA. Subgroups were examined based on the study design and pooled risk ratios (RRs) were estimated with 95% confidence intervals (CIs). For the meta-analysis, we focused on 11 out of 52 RCTs (Jadad ≥ 2) and found that CHM was more effective than placebo for HBeAg seroconversion when combined with WM (RR 4.67, 95% CI 1.36-15.98; P = 0.01; P = 39%); Radix Astragali was the most commonly used herb. Those that received CHM were more prone to adverse events; however, they were mild and reversible. The risk of bias was assessed with regards to blinding, incomplete outcome data and publication bias. It should be noted that, due to the poor methodological quality of the studies and the small number of RCTs, the results cannot fully support the use of CHM in the treatment of HBV carriers. To conclude, CHM may be used to treat HBV carriers, but rigorously designed RCTs with long-term follow-ups are required to further evaluate the benefits and safety of CHM.
Asunto(s)
Portador Sano/tratamiento farmacológico , Medicamentos Herbarios Chinos/uso terapéutico , Virus de la Hepatitis B , Hepatitis B Crónica/tratamiento farmacológico , HumanosRESUMEN
Calcitonin (CT) and calcitonin gene-related peptide (CGRP) are critical regulators of calcium balance and have extensive implications for vertebrate physiological processes. This study explores the CT and CGRP signaling systems in chickens through cloning and characterization of the chicken calcitonin receptor (CTR) and calcitonin receptor-like receptor (CLR), together with three receptor activity-modifying proteins (RAMPs). We illuminated the functional roles for chickens between the receptors examined alone and in RAMP-associated complexes using luciferase reporter assays. Chicken CTRs and CLRs stimulated the cAMP/PKA and MAPK/ERK signaling pathways, signifying their functional receptor status, with CT showing appreciable ligand activity at nanomolar concentrations across receptor combinations. Notably, it is revealed that chicken CLR can act as a functional receptor for CT without or with RAMPs. Furthermore, we uncovered a tissue-specific expression profile for CT, CGRP, CTR, CLR, and RAMPs in chickens, indicating the different physiological roles across various tissues. In conclusion, our data establish a clear molecular basis to reveal information on CT, CGRP, CTR, CLR, and RAMPs in chickens and contribute to understanding the conserved or divergent functions of this family in vertebrates.
RESUMEN
Somatostatin shows an anti-lipolytic effect in both chickens and ducks. However, its molecular mediator remains to be identified. Here, we report that somatostatin type 2 receptor (SSTR2) is expressed at a high level in chicken adipose tissue. In cultured chicken adipose tissue, the inhibition of glucagon-stimulated lipolysis by somatostatin was blocked by an SSTR2 antagonist (CYN-154086), supporting an SSTR2-mediated anti-lipolytic effect. Furthermore, a significant pro-proliferative effect was detected in SST28-treated immortalized chicken preadipocytes (ICP-1), and this cell proliferative effect may be mediated through the MAPK/ERK signaling pathway activated by SSTR2. In summary, our results demonstrate that SSTR2 may regulate adipose tissue development by affecting the number and volume of adipocytes in chickens.
RESUMEN
The laying hen is the spontaneous model of ovarian tumor. A comprehensive comparison based on RNA-seq from hens and women may shed light on the molecular mechanisms of ovarian cancer. We performed next-generation sequencing of microRNA and mRNA expression profiles in 9 chicken ovarian cancers and 4 normal ovaries, which has been deposited in GSE246604. Together with 6 public datasets (GSE21706, GSE40376, GSE18520, GSE27651, GSE66957, TCGA-OV), we conducted a comparative transcriptomics study between chicken and human. In the present study, miR-451, miR-2188-5p, and miR-10b-5p were differentially expressed in normal ovaries, early- and late-stage ovarian cancers. We also disclosed 499 up-regulated genes and 1,061 down-regulated genes in chicken ovarian cancer. The molecular signals from 9 cancer hallmarks, 25 Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways, and 369 Gene Ontology (GO) pathways exhibited abnormalities in ovarian cancer compared to normal ovaries via Gene Set Enrichment Analysis (GSEA). In the comparative analysis across species, we have uncovered the conservation of 5 KEGG and 76 GO pathways between chicken and human including the mismatch repair and ECM receptor interaction pathways. Moreover, a total of 174 genes contributed to the core enrichment for these KEGG and GO pathways were identified. Among these genes, the 22 genes were found to be associated with overall survival in patients with ovarian cancer. In general, we revealed the microRNA profiles of ovarian cancers in hens and updated the mRNA profiles previously derived from microarrays. And we also disclosed the molecular pathways and core genes of ovarian cancer shared between hens and women, which informs model animal studies and gene-targeted drug development.
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Pollos , MicroARNs , Neoplasias Ováricas , Transcriptoma , Pollos/genética , Femenino , Animales , Neoplasias Ováricas/genética , Neoplasias Ováricas/veterinaria , Humanos , MicroARNs/genética , MicroARNs/metabolismo , Enfermedades de las Aves de Corral/genética , Perfilación de la Expresión Génica/veterinaria , ARN Mensajero/genética , ARN Mensajero/metabolismoRESUMEN
BACKGROUND: Hepatitis C virus (HCV) is one of the major pathogens of liver diseases. Some studies have previously reported that miR-122 can stimulate replication or translation of HCV. However, the effects of HCV infection on miR-122 expression are not clear. The aim of this study was to investigate the effects of HCV core protein on the expression of miR-122 in a cell culture model. RESULTS: The miR-122 levels in Huh7.5.1 cells infected with HCV for different days or different HCV abundance were measured by real-time PCR. Significant decrease of miR-122 expression was found at late stage of infection and in the high-abundance group. Huh7.5.1 cells transfected with plasmid pEGFP-core or pEGFP were used to detect the effects of HCV core protein on miR-122 expression, the results showed that core protein could down-regulate the miR-122 expression level in a time- and dose- dependent manner, and reduced the susceptibility of Huh7.5.1 cell to HCV. CONCLUSIONS: Down-regulating miR-122 expression by HCV core protein may give a new insight into the interaction between HCV and miR-122 and chronic HCV infection.
Asunto(s)
Expresión Génica , Hepacivirus/patogenicidad , Interacciones Huésped-Patógeno , MicroARNs/biosíntesis , Proteínas del Núcleo Viral/metabolismo , Línea Celular , Perfilación de la Expresión Génica , Hepatocitos/virología , Humanos , Reacción en Cadena en Tiempo Real de la PolimerasaRESUMEN
OBJECTIVES: It has been found that bentonite particles (BPs) could induce the cyto-genotoxicity and oxidative stress in vitro, but these effects on population exposed to BPs remain unclear. The aim of the present study was to investigate whether the genetic damage and lipid peroxidation can be detected in workers occupationally exposed to organic BPs. METHODS: Sixty subjects were divided into three groups: (i) exposure group I consisted of 20 workers exposed to high concentrations of organic BPs in air; (ii) exposure group II were composed of 20 workers exposed to moderate concentrations of organic BPs in air; (iii) control group included 20 healthy unexposed subjects. Genetic damage was examined by comet assay and cytokinesis-block micronucleus cytome (CBMNcyt) assay. Lipid peroxidation was detected by malondialdehyde (MDA) assay. RESULTS: The % tail DNA, MDA, the frequencies of micronucleus (MNF), micronucleated cell (MCF), nucleoplasmic bridges (NPBs), nuclear buds (NBUDs), apoptotic cell rate (ACR) and necrotic cell rate (NCR) in two exposure groups were significantly higher than those in control group (P<0.01 or P<0.05). Moreover, the % tail DNA, MDA, MNF, MCF, NPBs, NBUDs, ACR and NCR in exposure group I with higher exposure level of organic BPs in air were significantly higher than those in exposure group II with lower level of organic BPs (P<0.01). The order of nuclear division index (NDI) was: exposure group IAsunto(s)
Bentonita/toxicidad
, Daño del ADN
, Peroxidación de Lípido
, Exposición Profesional
, Adulto
, Ensayo Cometa
, Relación Dosis-Respuesta a Droga
, Femenino
, Humanos
, Masculino
, Pruebas de Micronúcleos
, Persona de Mediana Edad
, Factores de Tiempo
RESUMEN
Dual siRNA against different regions of gene in hepatitis C virus (HCV) synergistically inhibited replication of HCV RNA. An HCV-infected cell model was established, and HCV RNA and core protein were detected by RT-PCR and Western blot, respectively. Four HCV-specific siRNAs (siCore, siNS3, siNS4B, siNS5B) were designed and transfected into HCV-infected Huh7.5.1 cells. The antiviral efficacies of the siRNAs were compared using real time PCR and agarose gel electrophoresis. HCV replication in infected cells was inhibited by IFNα-2b in a dose-dependent manner. Synergistic inhibition effects were achieved with combination treatment of any two of the siRNAs (siCore, siNS3 and siNS5B) at low doses (0.1 and 10 nM), as compared to single siRNA treatment (P < 0.05). Furthermore, CCK-8 assay showed no toxicity of the siRNAs to Huh7.5.1 cells. These findings indicate a promising new therapeutic approach for treatment of HCV.
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Antivirales/farmacología , Productos Biológicos/farmacología , Hepacivirus/crecimiento & desarrollo , ARN Interferente Pequeño/genética , ARN Interferente Pequeño/farmacología , Replicación Viral/efectos de los fármacos , Western Blotting , Línea Celular , Sinergismo Farmacológico , Hepatocitos/virología , Humanos , ARN Viral/biosíntesis , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Proteínas del Núcleo Viral/biosíntesisRESUMEN
The aim of the present study was to evaluate the genotoxic effects induced by native and active bentonite particles (BPs) on human B lymphoblast cells using comet assay and cytokinesis-block micronucleus (CBMN) assay in vitro. The cells were exposed to BPs at the concentrations of 30, 60, 120 and 240µg/ml for 24, 48 and 72h, respectively. The quartz contents of native and active BPs were 6.80±0.20 and 6.50±0.10%, respectively. Gypsum and DQ-12 quartz served as negative and positive controls. The results of comet assay showed that DNA damage induced by native and active BPs was significantly higher than that induced by gypsum control (P<0.05 or <0.01), and increased with exposure concentration and duration. When the cells were exposed to BPs at the doses of 120 and 240µg/ml for 72h, DNA damage induced by active BPs and native BPs was significantly higher than that induced by DQ-12 quartz (P<0.01), and DNA damage induced by active BPs enhanced significantly, as compared with native BPs (P<0.01). The results of CBMN assay demonstrated that both native BPs and active BPs could induce significant micronuclei, as compared with gypsum control (P<0.05 or <0.01). However, there was no significant difference of micronucleus frequency (MNF) among native BPs, active BPs and DQ-12 quartz. The water-soluble fractions from two kinds of BPs did not induce significant DNA damage and micronuclei. These findings indicated that the genotoxicity induced by active BPs and native BPs could be detected in comet assay and CBMN assay in vitro, the insoluble particle fractions from BPs may play a main role in the genotoxic effects induced by BPs.
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Linfocitos B/efectos de los fármacos , Bentonita/toxicidad , Ensayo Cometa/métodos , Pruebas de Micronúcleos/métodos , Bentonita/química , Línea Celular , Daño del ADN , Relación Dosis-Respuesta a Droga , Humanos , SolubilidadRESUMEN
OBJECTIVE: To investigate the inhibitive effects of small interfering RNA (siRNA) on hepatitis C virus (HCV) replication in cells infected by HCV in vitro. METHODS: The HCV RNA transcripts prepared by pFL-JC1 were transfected into Huh-7.5.1 cells. Na ve Huh-7.5.1 cells were incubated with the supernatants of transfected cells and the expression of HCV core protein in infected cells was detected by indirect immunofluorescence. The infected cells were transfected with 4, 40 and 200 nmol/L of NS5B siRNA for 24 h, 48 h and 72 h, respectively. The normal Huh-7.5.1 cells were transfected with 4, 40 and 200 nmol/L of NS5B siRNA. Group of blank, lipofectamine 2000, unrelated siRNA and IFNα-2b (1000 IU/ml) served as controls. The HCV RNA and PKR mRNA levels were examined by quantitative RT-PCR. RESULTS: The HCV core protein in HCV infected cells was detected. Compared with control groups, the HCV RNA levels in infected cells significantly decreased when transfected with 40 and 200 nmol/L of siRNA for 24 h; 4, 40 and 200 nmol/L of siRNA for 48 h and 72 h (P<0.05). The HCV RNA levels in infected cells treated with IFNα-2b (1000 IU/ml) for 24 h, 48 h and 72 h were significantly lower than those in control groups (P<0.05 or P<0.01). The PKR mRNA levels in Huh-7.5.1 cells transfected with siRNA of three concentrations did not have significant difference, as compared with control groups (P>0.05). CONCLUSION: siRNA against HCV NS5B region can effectively inhibit HCV replication in HCV infected cells, but can not activate the dsRNA-dependent protein kinase (PKR).
Asunto(s)
Hepacivirus/fisiología , ARN Interferente Pequeño/farmacología , Replicación Viral/efectos de los fármacos , Línea Celular Tumoral , Hepacivirus/efectos de los fármacos , Hepacivirus/genética , Humanos , Transfección , Proteínas no Estructurales Virales/genéticaRESUMEN
OBJECTIVE: To analyze the characteristics of pneumoconiosis cases in Zhejiang province and to provide the evidence for pneumoconiosis control and prevention measures in Zhejiang province. METHODS: The data of new pneumoconiosis cases were from national surveillance system of occupational disease in Zhejiang province during 2006-2009, and were analyzed for distribution, age, exposure duration, pneumoconiosis phases and enterprise types. RESULTS: During 2006-2009, 819 new pneumoconiosis cases (173, 157, 209 and 280 cases, respectively) were reported, 86.9% cases suffered from silicosis. Most of pneumoconiosis cases were distributed in Ningbo, Wenzhou areas and in building materials, machinery, coal, geological and mining, light industries and construction enterprise. The average ages of new pneumoconiosis cases were (47.8 +/- 10.0), (52.5 +/- 13.1), (55.5 +/- 11.2) and (55.9 +/- 12.2) years old, respectively and showed a significant increase trend (P<0.05). The average exposure duration of new pneumoconiosis cases were (12.4 +/- 8.6), (12.9 +/- 9.4), (12.4 +/- 8.6) and (15.7 +/- 10.0) years. The average exposure duration of phase I, phase II, phase III new pneumoconiosis cases were (14.3 +/- 9.87), (12.4 +/- 8.7) and (11.4 +/- 7.1) years, respectively and there were significant differences (P<0.05). CONCLUSION: New pneumoconiosis cases in Zhejiang province are increasing year by year, the main type of pneumoconiosis is silicosis, the distribution of pneumoconiosis cases is associated with the areas and enterprises, and the exposure duration of new pneumoconiosis cases is relatively shorter.
Asunto(s)
Enfermedades Profesionales/epidemiología , Neumoconiosis/epidemiología , Adulto , Anciano , China/epidemiología , Humanos , Persona de Mediana EdadRESUMEN
Congenital cataracts are the leading cause of childhood blindness. To date, surgical removal of cataracts is the only established treatment, but surgery is associated with multiple complications, which often lead to visual impairment. Therefore, mechanistic studies and drug-candidate screening have been intrigued by the aims of developing novel therapeutic strategies. However, these studies have been hampered by a lack of an appropriate human-disease model of congenital cataracts. Herein, we report the establishment of a human congenital cataract in vitro model through differentiation of patient-specific induced pluripotent stem cells (iPSCs) into regenerated lenses. The regenerated lenses derived from patient-specific iPSCs with known causative mutations of congenital cataracts (CRYBB2 [p. P24T] and CRYGD [p. Q155X]) showed obvious opacification that closely resembled that seen in patients' cataracts in terms of opacification severity and disease course accordingly, as compared with lentoid bodies (LBs) derived from healthy individuals. Increased protein aggregation and decreased protein solubility corresponding to the patients' cataract severity were observed in the patient-specific LBs and were attenuated by lanosterol treatment. Taken together, the in vitro model described herein, which recapitulates patient-specific clinical manifestations of congenital cataracts and protein aggregation in patient-specific LBs, provides a robust system for research on the pathological mechanisms of cataracts and screening of drug candidates for cataract treatment.
RESUMEN
In the present study, the cyto-genotoxic effects of 12 CSCs prepared from a diverse set of cigarettes on human B lymphoblastoid cells were compared using five in vitro assays. The cells were exposed to CSCs at doses of 2.5, 5.0, 7.5, 10.0, and 12.5 x 10(-3)cigarette/ml for 24h in neutral red uptake and CCK-8 assays, at doses of 1.0, 2.0, 3.0, 4.0, and 5.0 x 10(-3)cigarette/ml for 3h in cell apoptosis assay, at doses of 6.0, 8.0, 10.0, 12.0, and 14.0 x 10(-3)cigarette/ml for 4h in comet assay, and at doses of 1.0, 2.0, 4.0, 6.0, and 8.0 x 10(-3)cigarette/ml for 4h in micronucleus assay. The potency of 12 CSCs to induce corresponding toxic effects in each assay was calculated, and the correlations between the results in five assays were analyzed. Our investigation showed that the results of 12 CSCs in CCK-8 and cell apoptosis assays were positive, the results of 11 CSCs in neutral red uptake and comet assays were positive, and 9 CSCs could induce significantly the micronuclei in micronucleus assay. It was found that the potency to induce the cytotoxic effects among 12 CSCs ranged 9.694 folds in neutral red uptake assay and 6.43 folds in CCK-8 assay, the potency to induce cell apoptosis among 12 CSCs ranged 8.191 folds, the potency to induce DNA damage among 12 CSCs ranged 29.199 folds, the potency to induce micronuclei among 12 CSCs ranged 5.879 folds. Moreover, the good correlations were found between any two assays. It was suggested that the cyto-genotoxicity of CSCs from different brands of cigarettes varied greatly, comet assay might be a sensitive assay in assessing the genotoxicity induced by CSCs.