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BACKGROUND & AIMS: Protease-sensitive PNLIP variants were recently associated with chronic pancreatitis (CP) in European populations. The pathological mechanism yet remains elusive. Herein, we performed a comprehensive genetic and functional analysis of PNLIP variants found in a large Chinese cohort, aiming to further unravel the enigmatic association of PNLIP variants with CP. METHODS: All coding and flanking intronic regions of the PNLIP gene were analyzed for rare variants by targeted next-generation sequencing in 1082 Chinese CP patients and 1196 controls. All novel missense variants were subject to analysis of secretion, lipase activity, and proteolytic degradation. One variant was further analyzed for its potential to misfold and induce endoplasmic reticulum (ER) stress. p.F300L, the most common PNLIP variant associated with CP, was used as a control. RESULTS: We identified 12 rare heterozygous PNLIP variants, with 10 being novel. The variant carrier frequency did not differ between the groups. Of them, only the variant p.A433T found in a single patient was considered pathologically relevant. p.A433T exhibited increased susceptibility to proteolytic degradation, which was much milder than p.F300L. Interestingly, both variants exhibited an increased tendency to misfold, leading to intracellular retention as insoluble aggregates, reduced secretion, and elevated ER stress. CONCLUSIONS: Our genetic and functional analysis of PNLIP variants identified in a Chinese CP cohort suggests that the p.A433T variant and the previously identified p.F300L variant are not only protease-sensitive but also may be potentially proteotoxic. Mouse studies of the PNLIP p.F300L and p.A433T variants are needed to clarify their role in CP.
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Povo Asiático , Predisposição Genética para Doença , Pancreatite Crônica , Humanos , Pancreatite Crônica/genética , Masculino , Povo Asiático/genética , Feminino , Estudos de Coortes , Pessoa de Meia-Idade , Adulto , Lipase/genética , Estresse do Retículo Endoplasmático/genética , China/epidemiologia , Mutação de Sentido Incorreto , Idoso , Variação Genética , População do Leste AsiáticoRESUMO
BACKGROUND: As of September 2023, more than 1,000 cases of monkeypox (mpox) have been reported in China. Based on the available evidence, men who have sex with men (MSM) are at high risk for mpox infection. This study aimed to analyses the self-reported infection status, knowledge, attitude and influencing factors of monkeypox among MSM in Jiaxing City, China. METHODS: A web-based cross-sectional survey was conducted in September 2023 to gather data on participants' socio-demographic profiles, mpox-related knowledge, sexual behavior characteristics, and other potentially related information to mpox knowledge. Multivariate regression modeling was employed to analyze the factors influencing the level of mpox-related knowledge. RESULTS: A total of 562 MSM were recruited; 4.3% self-reported being HIV-positive, 83.3% of respondents had heard of mpox, and 2.3% of them reported having suspected symptoms. 89.7% of respondents were willing to be vaccinated against mpox, but only 24.8% had a high level of knowledge about mpox. The main factors influencing knowledge of mpox were education level, household registration, homosexual anal intercourse in the past 6 months, and taking the HIV pre-exposure prophylaxis (PrEP). CONCLUSIONS: Knowledge of mpox among MSM living in the Jiaxing area needs to be enhanced, but willingness to get vaccinated is high. Educational level, household location, sexual behavior and PrEP use have important effects on knowledge of mpox. Individuals exhibiting symptoms indicative of suspected mpox had a diminutive consultation frequency, and it is imperative to augment screening efforts for mpox symptoms within specific demographic groups to prevent the underreporting of mpox cases.
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Conhecimentos, Atitudes e Prática em Saúde , Homossexualidade Masculina , Mpox , Autorrelato , Humanos , Masculino , China/epidemiologia , Adulto , Homossexualidade Masculina/estatística & dados numéricos , Homossexualidade Masculina/psicologia , Estudos Transversais , Adulto Jovem , Pessoa de Meia-Idade , Mpox/epidemiologia , Adolescente , Fatores de RiscoRESUMO
OBJECTIVE: Increasing evidence implicates mutation-induced protein misfolding and endoplasm reticulum (ER) stress in the pathophysiology of chronic pancreatitis (CP). The paucity of animal models harbouring genetic risk variants has hampered our understanding of how misfolded proteins trigger CP. We previously showed that pancreatic triglyceride lipase (PNLIP) p.T221M, a variant associated with steatorrhoea and possibly CP in humans, misfolds and elicits ER stress in vitro suggesting proteotoxicity as a potential disease mechanism. Our objective was to create a mouse model to determine if PNLIP p.T221M causes CP and to define the mechanism. DESIGN: We created a mouse model of Pnlip p.T221M and characterised the structural and biochemical changes in the pancreas aged 1-12 months. We used multiple methods including histochemistry, immunostaining, transmission electron microscopy, biochemical assays, immunoblotting and qPCR. RESULTS: We demonstrated the hallmarks of human CP in Pnlip p.T221M homozygous mice including progressive pancreatic atrophy, acinar cell loss, fibrosis, fatty change, immune cell infiltration and reduced exocrine function. Heterozygotes also developed CP although at a slower rate. Immunoblot showed that pancreatic PNLIP T221M misfolded as insoluble aggregates. The level of aggregates in homozygotes declined with age and was much lower in heterozygotes at all ages. The Pnlip p.T221M pancreas had increased ER stress evidenced by dilated ER, increased Hspa5 (BiP) mRNA abundance and a maladaptive unfolded protein response leading to upregulation of Ddit3 (CHOP), nuclear factor-κB and cell death. CONCLUSION: Expression of PNLIP p.T221M in a preclinical mouse model results in CP caused by ER stress and proteotoxicity of misfolded mutant PNLIP.
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Pancreatite Crônica , Camundongos , Humanos , Animais , Pancreatite Crônica/genética , Pâncreas/metabolismo , Células Acinares/metabolismo , Estresse do Retículo Endoplasmático/genética , Resposta a Proteínas não Dobradas , Chaperona BiP do Retículo EndoplasmáticoRESUMO
Salmonella is a primary cause of foodborne diseases, and the increasing prevalence of mcr-1-carrying plasmids, which confer colistin resistance to Salmonella, poses significant global health concerns. As the frequency of occurrence of the mcr-1 gene is increasing globally, we studied the prevalence of mcr-1 in clinical Salmonella isolates by analyzing 195 clinical strains isolated in 2020. Of the 195 Salmonella isolates, 41 isolates were resistant to colistin. We found mcr-1 in two strains (Salmonella Typhimurium ZJJX20006 and Salmonella Kentucky ZJJX20014), which we analyzed in detail via whole-genome sequencing and antibiotic susceptibility testing. Two strains displayed resistance to ampicillin, ampicillin-sulbactam, tetracycline, chloramphenicol, and cotrimoxazole, while ZJJX20006 displayed resistance to colistin and ZJJX20014 was sensitive. Genomic analysis revealed that these strains had plasmid-encoded mcr-1 in IncHI2 plasmids, which were not similar to the mcr-1-IncX4 identified in 2016. These two strains also harbored other drug resistance genes, including blaOXA-1 and blaCTX-M-14. Our findings may help clarify the molecular mechanisms of mcr-1 dissemination among Salmonella strains in Jiaxing City and offer insights into the evolution of mcr-1 in Salmonella.
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The imbalance that occurs in bone remodeling induced by irradiation (IR) is the disruption of the balance between bone formation and bone resorption. In this study, primary osteocytes (OCYs) of femoral and tibial origin were cultured and irradiated. It was observed that irradiated OCY showed extensive DNA damage, which led to the initiation of a typical phenotype of cellular senescence, including the secretion of senescence-associated secretory phenotype (SASP), especially the C-C motif chemokine ligand 5 (CCL5). In order to explore the regulation of osteoclastogenic potential by IR-induced senescent OCYs exocytosis factor CCL5, the conditioned medium (CM) of OCYs was co-cultured with RAW264.7 precursor cells. It was observed that in the irradiated OCY co-cultured group, the migration potential increased compared with the vehicle culture group, accompanied by an enhancement of typical mature OCs; the expression of the specific function of enzyme tartrate-resistant acid phosphatase (TRAP) increased; and the bone-destructive function was enhanced. However, a neutralizing antibody to CCL5 could reverse the extra-activation of osteoclastogenesis. Accordingly, the overexpression of p-STAT3 in irradiated OCY was accompanied by CCL5. It was concluded that CCL5 is a potential key molecule and the interventions targeting CCL5 could be a potential strategy for inhibiting osteoclastogenesis and restoring bone remodeling.
Assuntos
Reabsorção Óssea , Osteogênese , Humanos , Remodelação Óssea , Reabsorção Óssea/metabolismo , Senescência Celular/genética , Quimiocina CCL5/genética , Quimiocina CCL5/metabolismo , Ligantes , Osteoclastos/metabolismo , Osteogênese/genética , Ligante RANK/metabolismo , Animais , CamundongosRESUMO
Background and Objectives: Damage to normal bone tissue following therapeutic irradiation (IR) represents a significant concern, as IR-induced bone microenvironment disruption can cause bone loss and create a more favorable environment for tumor metastases. The aim of the present study was to explore the cellular regulatory mechanism of IR-induced bone microenvironment disruption to effectively prevent radiotherapy-associated adverse effects in the future. Materials and Methods: In this study, a mouse model of local IR was established via local irradiation of the left hind limb of BALB/c mice with 12 Gy X-rays, and an in vitro osteocyte (OCY) model was established by exposing osteocyte-like MLO-Y4 cells to 2, 4, and 8 Gy irradiation to analyze multicellular biological injuries and cellular senescence. Small interfering RNA (siRNA) transfection at the cellular level and a selective antagonist intervention C-176 at the animal level were used to explore the potential role of the stimulator of interferon genes (STING) on IR-induced bone microenvironment disruption. Results: The results showed that 12 Gy local IR induces multicellular dysfunction, manifested as ascension of OCYs exfoliation, activation of osteoclastogenesis, degeneration of osteogenesis and fate conversion of adipogenesis, as well as cellular senescence and altered senescence-associated secretory phenotype (SASP) secretion. Furthermore, the expression of STING was significantly elevated, both in the primary OCYs harvested from locally irradiated mice and in vitro irradiated MLO-Y4 cells, accompanied by the markedly upregulated levels of phosphorylated TANK-binding kinase 1 (P-TBK1), RANKL and sclerostin (SOST). STING-siRNA transfection in vitro restored IR-induced upregulated protein expression of P-TBK1 and RANKL, as well as the mRNA expression levels of inflammatory cytokines, such as IL-1α, IL-6 and NF-κB, accompanied by the alleviation of excessive osteoclastogenesis. Finally, administration of the STING inhibitor C-176 mitigated IR-induced activation of osteoclastogenesis and restraint of osteogenesis, ameliorating the IR-induced biological damage of OCYs, consistent with the inhibition of P-TBK1, RANKL and SOST. Conclusions: The STING-P-TBK1 signaling pathway plays a crucial role in the regulation of the secretion of inflammatory cytokines and osteoclastogenesis potential in IR-induced bone microenvironment disruption. The selective STING antagonist can be used to intervene to block the STING pathway and, thereby, repair IR-induced multicellular biological damage and mitigate the imbalance between osteoclastogenesis and osteoblastgenesis.
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Osso e Ossos , Transdução de Sinais , Animais , Camundongos , Osso e Ossos/metabolismo , Citocinas , NF-kappa B/metabolismo , Osteogênese , Transdução de Sinais/fisiologia , Camundongos Endogâmicos BALB CRESUMO
This study was to develop a multiplex fluorescent PCR for Shigella detection and species identification. Five primer pairs for Shigella detection and species identification were designed by Primer Premier 5.0. The multiplex fluorescent PCR was optimized by varying single parameter while other parameters were maintained. The multiplex fluorescent PCR assay could correctly detect Shigella and identify four Shigella species with a detection limits of 10 pg genomic DNA per reaction. Testing different strains and clinical samples confirmed the sensitivity and specificity of the multiplex fluorescent PCR. The newly developed multiplex fluorescent PCR assay is simple, sensitive and specific for Shigella detection and species identification. It has a potential to be used in routine Shigella detection and species identification in clinical laboratories.
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Fluorescência , Reação em Cadeia da Polimerase Multiplex/métodos , Shigella/classificação , Shigella/isolamento & purificação , Genes Bacterianos , Humanos , Sensibilidade e Especificidade , Shigella/genéticaRESUMO
Low hemoglobin (Hb) level or anemia is associated with osteoporosis and bone fracture. Cadmium (Cd) and lead (Pb) exposure are also risk factors of osteoporosis and anemia. However, the role of anemia in Cd/Pb related bone loss remains unclear. The aim of present study was to investigate the association between Hb level and bone loss in a population with environmental lead and cadmium exposure. One hundred and ninety-four women and 108 men with different levels of Cd/Pb exposure were included in our study. The Cd/Pb exposure was determined using graphite-furnace atomic absorption spectrometry. Forearm bone mineral density (BMD) was determined by peripheral dual-energy X-ray absortiometry. Hb concentration was determined using an automatic blood cellcounter. A logistic model was established to predict the risk of osteoporosis. The BMDs of women that had the highest quartile BCd and BPb were markedly lower than that with the lowest quartile (p < 0.05). The BMD and the prevalence of osteoporosis in men with anemia were lower and higher than that with normal Hb (p < 0.05), respectively. In men, age, BPb and anemia were independent risk factors for osteoporosis. The odds ratio (OR) of men with anemia was 11.28 (95%confidence interval (CI):1.94-65.54) and 19.56 (95%CI: 2.98-128.78) compared to those with normal Hb after adjusting for potential cofounders. No such association was found in women. The area under the curve was 0.88 (95%CI: 0.82-0.96) in predicting osteoporosis using the logistic model in men. Linear discriminant analysis also showed that 90.7% of osteoporosis was correctly classified. Our data show that anemia is associated with incident of osteoporosis in men but not in women that environmentally exposed to Pb and Cd.
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Cádmio , Osteoporose , Densidade Óssea , Cádmio/análise , Cádmio/toxicidade , China/epidemiologia , Exposição Ambiental/análise , Feminino , Hemoglobinas/análise , Humanos , Chumbo/análise , Chumbo/toxicidade , Masculino , Osteoporose/induzido quimicamente , Osteoporose/epidemiologiaRESUMO
The association between internal cadmium exposure and chronic kidney disease (CKD) has been investigated before. However, few studies have shown the association between dietary cadmium intake and CKD. In this study, we show the association between life-time dietary cadmium intake and CKD based on a follow-up study. At baseline, we collected blood and urine samples for assays of cadmium and renal effect biomarkers. A questionnaire and food survey was given to each subject to collect diet and lifestyle information for the estimation of cadmium intake. Dietary cadmium, cadmium in blood and urine were regarded as exposure markers. Life-time dietary cadmium intake was estimated based on an individual's daily cadmium intake and exposure time. At follow-up, 467 persons (163 men and 304 women) were finally included. CKD at follow-up was considered if the estimated glomerular filtration rate (eGFR) was less than 60 mL/min/1.73 m2. The eGFR level in subjects in the highest quartile of total dietary cadmium intake (>9.34 g) was significantly lower than in those with a moderate or low intake (p < 0.01). eGFR was negatively associated with total dietary cadmium intake (ß = -0.42, 95% confidence interval (CI): -0.77 to -0.07) after adjustment with confounders. Logistic regression further showed that the risk of CKD in subjects with a high total dietary cadmium intake (>2.2 g) was higher than in those with a low intake (odds ratio (OR) = 18.16, 95%CI: 1.75-188.85). A similar association was found between the baseline urinary albumin (UALB) level and CKD incidence. A predictive model based on UALB and life-time dietary cadmium intake showed an acceptable performance (the area under the curve was 0.77 (95%CI: 0.65-0.88)). Our data show that high dietary cadmium exposure was associated with CKD after controlling for renal tubular dysfunction and internal cadmium exposure.
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Cádmio/sangue , Exposição Dietética/estatística & dados numéricos , Poluentes Ambientais/sangue , Insuficiência Renal Crônica/epidemiologia , Adulto , Dieta , Feminino , Seguimentos , Taxa de Filtração Glomerular , Humanos , Incidência , Rim/fisiopatologia , Pessoa de Meia-Idade , Razão de Chances , Oryza , Fatores de RiscoRESUMO
Cellular senescence and its senescence-associated secretory phenotype (SASP) are widely regarded as promising therapeutic targets for aging-related diseases, such as osteoporosis. However, the expression pattern of cellular senescence and multiple SASP secretion remains unclear, thus leaving a large gap in the knowledge for a desirable intervention targeting cellular senescence. Therefore, there is a critical need to understand the molecular mechanism of SASP secretion in the bone microenvironment that can ameliorate aging-related degenerative pathologies including osteoporosis. In this study, osteocyte-like cells (MLO-Y4) were induced to cellular senescence by 2 Gy γ-rays; then, senescence phenotype changes and adverse effects of SASP on bone marrow mesenchymal stem cell (BMSC) differentiation potential were investigated. The results revealed that 2 Gy irradiation could hinder cell viability, shorten cell dendrites, and induce cellular senescence, as evidenced by the higher expression of senescence markers p16 and p21 and the elevated formation of senescence-associated heterochromatin foci (SAHF), which was accompanied by the enhanced secretion of SASP markers such as IL-1α, IL-6, MMP-3, IGFBP-6, resistin, and adiponectin. When 0.8 µM JAK1 inhibitors were added to block SASP secretion, the higher expression of SASP was blunted, but the inhibition in osteogenic and adipogenic differentiation potential of BMSCs co-cultured with irradiated MLO-Y4 cell conditioned medium (CM- 2 Gy) was alleviated. These results suggest that senescent osteocytes can perturb BMSCs' differential potential via the paracrine signaling of SASP, which was also demonstrated by in vivo experiments. In conclusion, we identified the SASP factor partially responsible for the degenerative differentiation of BMSCs, which allowed us to hypothesize that senescent osteocytes and their SASPs may contribute to radiation-induced bone loss.
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Reabsorção Óssea/patologia , Diferenciação Celular , Senescência Celular , Raios gama/efeitos adversos , Células-Tronco Mesenquimais/patologia , Osteócitos/patologia , Comunicação Parácrina , Animais , Reabsorção Óssea/etiologia , Reabsorção Óssea/metabolismo , Masculino , Células-Tronco Mesenquimais/efeitos da radiação , Camundongos , Camundongos Endogâmicos BALB C , Osteócitos/efeitos da radiação , OsteogêneseRESUMO
The role of cellular senescence induced by radiation in bone loss has attracted much attention. As one of the common complications of anticancer radiotherapy, irradiation-induced bone deterioration is common and clinically significant, but the pathological mechanism has not been elucidated. This study was performed to explore the cellular senescence and senescence-associated secretory phenotype (SASP) induction of bone marrow-derived mesenchymal stem cells (BMSCs) by irradiation and its role in osteogenic differentiation dysfunction. It was observed that irradiated BMSCs lost typical fibroblast-like morphology, exhibited suppressed viability and differentiation potential accompanied with senescence phenotypes, including an increase in senescence-associated ß-galactosidase (SA-ß-gal) staining-positive cells, and upregulated senescence-related genes p53/p21, whereas no changes happened to p16. Additionally, DNA damage γ-H2AX foci, G0/G1 phase of cell cycle arrest, and cellular and mitochondrial reactive oxygen species (ROS) increased in an irradiation dose-dependent manner. Meanwhile, the JAK1/STAT3 pathway was activated and accompanied by an increase in SASP secretion, such as IL-6, IL-8, and matrix metalloproteinase-9 (MMP9), whereas 0.8 µM JAK1 inhibitor (JAKi) treatment effectively inhibited the JAK pathway and SASP production. Furthermore, conditioned medium (CM) from irradiation-induced senescent (IRIS) BMSCs exhibited a markedly reduced ability in osteogenic differentiation and marker gene expression of osteoblasts, whereas CM with JAKi intervention may effectively improve these deterioration effects. In conclusion, irradiation could provoke BMSC senescence and SASP secretion and further aggravate osteogenic differentiation dysfunction via paracrine signaling, whereas SASP targeting may be a possible intervention strategy for alleviating irradiation-induced bone loss.
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Diferenciação Celular/genética , Senescência Celular/genética , Células-Tronco Mesenquimais/citologia , Osteogênese/genética , Reabsorção Óssea/genética , Reabsorção Óssea/terapia , Pontos de Checagem do Ciclo Celular/genética , Proliferação de Células/genética , Senescência Celular/efeitos da radiação , Dano ao DNA/efeitos da radiação , Regulação da Expressão Gênica no Desenvolvimento/efeitos da radiação , Histonas/genética , Humanos , Janus Quinase 1/genética , Células-Tronco Mesenquimais/efeitos da radiação , Mitocôndrias/genética , Mitocôndrias/efeitos da radiação , Comunicação Parácrina/genética , Radiação , Espécies Reativas de Oxigênio/metabolismo , Fator de Transcrição STAT3/genética , Transdução de Sinais/efeitos da radiaçãoRESUMO
Cadmium exposure is associated with renal dysfunction. However, the outcome of renal function in subjects who have had a reduction in cadmium exposure for years has not been completely clarified, particularly for individuals with normal baseline renal function. In this study, we used a nomogram model to predict renal dysfunction after a reduction in cadmium exposure in subjects with normal baseline renal function. In 1998, a survey was performed in 790 subjects living in control and cadmium-polluted areas. A total of 497 subjects was followed up in 2006. 404 subjects with normal baseline urinary ß2-microglobulin (UBMG), 373 subjects with normal baseline urinary N-acetyl-ß-d-glucosaminidase (UNAG) and 407 subjects with normal baseline urinary albumin (UALB) were included in this analysis. Cadmium in the blood (BCd) and urine (UCd) was detected using graphite-furnace atomic absorption spectrometry. A logistic regression model was used to identify potential predicting factors of renal function at follow-up. Nomograms were developed based on those predictive factors. Bootstrap self-sampling, calibration curves and receiver operating characteristic (ROC) curves were performed to quantify our modeling strategy. Adjusted and unadjusted logistic regression models both showed that age, BCd and UBMG or UNAG at baseline were independent risk factors for renal tubular dysfunction. Baseline age, sex, BCd and UNAG or UBMG or UALB were used to construct the nomogam. The internal validation showed that the C-index was 0.70-0.74 for predicting renal dysfunction. The area under the curve of the nomogram was 0.70-0.74. Decision curve analysis verified the predictive value of the nomogram. A nomogram may be a useful risk assessment tool for predicting the progress of renal function in a cadmium-exposed population with normal baseline kidney function.
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Cádmio/sangue , Exposição Ambiental/análise , Poluentes Ambientais/sangue , Nefropatias/epidemiologia , Nomogramas , Acetilglucosaminidase/urina , Adulto , China , Exposição Ambiental/efeitos adversos , Feminino , Seguimentos , Humanos , Nefropatias/induzido quimicamente , Nefropatias/urina , Testes de Função Renal , Modelos Logísticos , Masculino , Pessoa de Meia-Idade , Medição de Risco , Microglobulina beta-2/urinaRESUMO
This study focused on using pH as a single indicator to evaluate/control the performance of the nitritation system under the influence of three major operational parameters, and a total of fifteen batch tests were conducted. Results indicated that there were important interactions among different operational parameters and pH in the nitritation system; it was possible to propose the optimal nitritation operation scheme to compensate for negative changes in operational parameters. The optimal carbon to nitrogen (C/N) ratio was kept at 2.0 to ensure efficient removal of ammonium. The reaction time was the lowest (150 min) with the temperature = 20 °C, C/N = 0, and sludge/water ratio = 1:1. However, the C/N ratio could be adjusted to close to zero by reducing the temperature to about 10 °C, weakening the heterotrophic bacteria, and supplying sufficient biomass. The C/N ratio and sludge/water ratio could also be set at 4.0 and 1:3 respectively to deal with the impact of low temperature and organic matter. Results of this study might be useful to explain the optimal conditions and process control schemes with pH as a single indicator.
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Compostos de Amônio , Reatores Biológicos , Concentração de Íons de Hidrogênio , Nitritos , Nitrogênio , EsgotosRESUMO
Irradiation-induced bone loss is widely reported, especially in radiotherapy-induced osteoporosis. In addition to the mechanism of osteogenesis inhibition and osteoclastogenesis promotion, the regulation effect of osteocytes, which also send signals to modulate osteoclastogenesis, should be elucidated. In this study, the effect of irradiation on osteocyte and its accommodation to osteoclastogenesis via the release of high mobility group box 1 (HMGB1) was explored. Furthermore, the control response of HMGB1 inhibitor on receptor activator of nuclear factor-κB ligand (RANKL) and osteoprotegerin (OPG) expression in osteocyte and osteocyte-induced osteoclastogenesis was assessed. It was observed that irradiated osteocyte-like MLO-Y4 cells exhibited polygonal-shaped morphological changes and shortened dendrites, inhibited cell viability and induced cellular apoptosis, along with the reduction in dendritic E11 protein/messenger RNA expression at a doses of 4 Gy. Additionally, the secretion of HMGB1 in supernatants was promoted, accompanied by the decreased OPG and elevated RANKL expression. When the RAW264.7 cells were cocultured with irradiated MLO-Y4 cells or its conditioned medium, enhanced migration and differentiation of osteoclast precursor was observed, and this difference was alleviated with anti-HMGB1 neutralizing antibody. In conclusion, this study demonstrated that irradiation deteriorated osteocytes' potential to promote recruitment and differentiation of osteoclast precursor via stimulating HMGB1 release and subsequent elevation of RANKL/OPG level. This study will assist in designing the intervention programs for irradiation-induced bone loss.
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Proteína HMGB1/metabolismo , Osteoclastos/metabolismo , Osteócitos/efeitos dos fármacos , Osteogênese/efeitos dos fármacos , Animais , Diferenciação Celular/efeitos dos fármacos , Diferenciação Celular/fisiologia , Linhagem Celular , Técnicas de Cocultura/métodos , Meios de Cultivo Condicionados/metabolismo , Fator Estimulador de Colônias de Macrófagos/metabolismo , Osteoclastos/efeitos dos fármacos , Osteócitos/metabolismo , Ligante RANK/metabolismoRESUMO
Both cadmium (Cd) and lead (Pb) exposure can induce kidney damage. However, the effects of combined exposure to Cd and Pb on renal function at environmental levels have not been fully clarified. In this study we investigated the renal function in a Chinese population co-exposed to Cd and Pb. A total of 331 subjects (215 women and 116 men), living in either a control or a polluted area, were included in this study. Cd and Pb in blood and urine (BCd, BPb, UCd, and UPb), and kidney effect markers including urinary N-acetyl-ß-D-glucosaminidase (UNAG) and estimated glomerular filtration rate (eGFR), were determined, and the association between exposure markers and renal effect biomarkers were analyzed. The exposure levels in the polluted area were significantly higher than in the control area (all pâ¯<â¯0.01). The eGFR of subjects in the polluted area was decreased compared with that in the control area (pâ¯<â¯0.01). The subjects with high BCd/BPb (BCd ≥ 2⯵g/L, BPb ≥ 100⯵g/L) or high UCd/UPb (UCd ≥ 3⯵g/g creatinine, UPb ≥ 10⯵g/g creatinine) showed higher UNAG and UALB levels compared with other subgroups (pâ¯<â¯0.01). The probability of having elevated UNAG in subjects with high BCd/BPb was greater than those with low BCd/BPb [odds ratio (OR) =â¯2.6, 95% confidence interval (CI): 1.4-4.7), low BCd/high BPb (OR =3.1, 95% CI: 1.4-6.6), and high BCd/low BPb (OR = 1.7, 95% CI: 0.9-3.2). The OR of subjects with low UCd and high UPb, high UCd and low UPb, and high UCd/UPb were 2.9 (95% CI: 1.4-5.7), 3.3 (95% CI: 1.5-7.2), and 7.7 (95% CI: 4.0-14.7), respectively, compared with those with low UCd/UPb. The risk of decrease in eGFR was also higher in subjects with high UCd/UPb than for those with low UCd/UPb (OR = 7.2, 95% CI: 0.8-62.2). Our data demonstrate that Cd and Pb exposure, alone or in combination, are associated with renal impairment. In addition, co-exposure to Pb and Cd propagates the renal tubular dysfunction compared with Cd or Pb exposure alone.
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Cádmio/toxicidade , Poluentes Ambientais/toxicidade , Rim/efeitos dos fármacos , Chumbo/toxicidade , Acetilglucosaminidase/urina , Adulto , Idoso , Idoso de 80 Anos ou mais , Povo Asiático , Biomarcadores/urina , Cádmio/sangue , Cádmio/urina , Exposição Ambiental/efeitos adversos , Poluentes Ambientais/sangue , Poluentes Ambientais/urina , Feminino , Taxa de Filtração Glomerular/efeitos dos fármacos , Humanos , Rim/fisiologia , Chumbo/sangue , Chumbo/urina , Masculino , Pessoa de Meia-Idade , Razão de ChancesRESUMO
Ionizing radiation-induced bone loss is a potential health concern in radiotherapy, occupational exposure, and astronauts. Although impaired bone vasculature and reduced proliferation of bone-forming osteoblasts has been implicated in this process, it has not been clearly characterized that whether radiation affects the growth of bone-resorbing osteoclasts. The molecular crosstalk between different cell populations in the skeletal system has not yet been elucidated in detail, especially between the increased bone resorption at early stage of post-irradiation and bone marrow-derived endothelial progenitor cells (BM-EPCs). In order to further understand the mechanisms involved in radiation-induced bone loss at the cellular level, we assessed the effects of irradiation on angiogenesis of BM-EPCs and osteoclastogenesis of receptor activator for nuclear factor-κB ligand (RANKL)-stimulated RAW 264.7 cells and crosstalk between these cell populations. We herein found significantly dysfunction of BM-EPCs in response to irradiation at a dose of 2 Gy, including inhibited proliferation, migration, tube-forming abilities, and downregulated expression of pro-angiogenesis vascular endothelial growth factors A (VEGF A). Meanwhile, we observed that irradiation promoted osteoclastogenesis of RANKL-stimulated RAW 264.7 cells directly or indirectly. These results provide quantitative evidences of irradiation induced osteoclastogenesis at a cellular level, and strongly suggest the involvement of osteoclastogenesis, angiogenesis and crosstalk between bone marrow cells in the radiation-induced bone loss. This study may provide new insights for the early diagnosis and intervention of bone loss post-irradiation.
Assuntos
Células da Medula Óssea/efeitos da radiação , Osteoblastos/efeitos da radiação , Osteoclastos/efeitos da radiação , Osteogênese/fisiologia , Indutores da Angiogênese/farmacologia , Animais , Células da Medula Óssea/metabolismo , Reabsorção Óssea/metabolismo , Diferenciação Celular/fisiologia , Fator Estimulador de Colônias de Macrófagos/metabolismo , Macrófagos/metabolismo , Masculino , Osteoblastos/metabolismo , Osteoclastos/metabolismo , Ligante RANK/metabolismo , Ratos Sprague-DawleyRESUMO
Bone homeostasis depends on the resorption of bones by osteoclasts and formation of bones by the osteoblasts. Imbalance of this tightly coupled process can cause diseases such as osteoporosis. Thus, the mechanisms that regulate communication between osteoclasts and osteoblasts are critical to bone cell biology. It has been shown that osteoblasts and osteoclasts can communicate with each other through direct cell-cell contact, cytokines, and extracellular matrix interaction. Osteoblasts can affect osteoclast formation, differentiation, or apoptosis through several pathways, such as OPG/RANKL/RANK, RANKL/LGR4/RANK, Ephrin2/ephB4, and Fas/FasL pathways. Conversely, osteoclasts also influence formation of bones by osteoblasts via the d2 isoform of the vacuolar (H+) ATPase (v-ATPase) V0 domain (Atp6v0d2), complement component 3a, semaphorin 4D or microRNAs. In addition, cytokines released from the resorbed bone matrix, such as TGF-ß and IGF-1, also affect the activity of osteoblasts. Drugs could be developed by enhancing or restricting some of these interactions. Several reviews have been performed on the osteoblast-osteoclast communication. However, few reviews have shown the research advances in the recent years. In this review, we summarized the current knowledge on osteoblast-osteoclast communication.
Assuntos
Apoptose , Comunicação Celular , Diferenciação Celular , Osteoblastos/metabolismo , Osteoclastos/metabolismo , Osteogênese , Transdução de Sinais , Animais , Humanos , Osteoblastos/citologia , Osteoclastos/citologiaRESUMO
The tolerable dietary intake of cadmium was recommended at provisional tolerable monthly intake of 25 µg kg-1 body weight. However, several studies indicated that this tolerable level should be re-evaluated for sufficient health protection. In this study, we show the reference levels of dietary cadmium intake for renal dysfunction by using a benchmark dose (BMD) approach. A total of 790 subjects (302 men and 488 women) living in control and cadmium-polluted areas were included. The dietary cadmium intake was estimated by a food survey. Blood cadmium, urinary cadmium and renal function markers (microalbuminuria, N-acetyl-ß-d-glucosaminidase [NAG] and its isoform B [NAGB], ß2 -microglobulin and retinol binding protein) in urine were measured. We calculated the 95% lower confidence bounds of BMD (BMDLs) of cumulative cadmium intake. In control and two polluted areas, the median cumulative cadmium intake was 0.5, 2.1 and 11.1 g. The odds ratio of the intermediate (1.0-3.0 g), second highest (3.0-11.0 g) and the highest cumulative cadmium intake (>11.0 g) compared with the lowest cumulative cadmium intake (<1.0 g) were 2.8 (95% CI: 1.4-5.8), 8.1 (95% CI: 3.8-17.2) and 11.4 (95% CI: 6.5-26.4) for urinary NAG and 6.6 (95% CI: 3.2-13.8), 14.8 (95% CI: 6.8-32.2) and 22.5 (95% CI: 10.7-47.5) for urinary NAGB. The BMDLs of cumulative cadmium intake were 1.1-1.2 g (benchmark response [BMR] = 5%) for urinary NAG, and were 0.7-0.9 g (BMR = 5%) for urinary NAGB, and were 1.3-1.4 g (BMR = 5%) for urinary ß2 -microglobulin. The BMDLs of cumulative cadmium intake in a Chinese population were lower than the critical standard previously reported. Further evaluations are needed for sufficient health protection.
Assuntos
Cádmio , Exposição Dietética/análise , Poluentes Ambientais , Rim/fisiopatologia , Adulto , Benchmarking , Biomarcadores/análise , Cádmio/sangue , Cádmio/urina , China , Exposição Dietética/efeitos adversos , Relação Dose-Resposta a Droga , Poluentes Ambientais/sangue , Poluentes Ambientais/urina , Feminino , Humanos , Rim/efeitos dos fármacos , Testes de Função Renal , Masculino , Pessoa de Meia-Idade , Razão de ChancesRESUMO
Osteoclasts, derived from hemopoietic progenitors of the monocyte/macrophage lineage, have a unique role in bone resorption, and are considered a potential therapeutic target in the treatment of such pathologic bone diseases as osteoporosis, rheumatoid arthritis, and periodontitis. In the present study, we demonstrate that curcumol, one of the major components of the essential oil of Rhizoma Curcumae, exhibits an inhibitory effect on receptor activator of nuclear factor kappaB ligand (RANKL)-induced osteoclast differentiation with both bone marrow-derived macrophages and RAW264.7 cells in a dose-dependent manner. In addition, RANKL-induced mRNA expression of osteoclast-specific genes, such as tartrate-resistant acid phosphatase, calcitonin receptor, and cathepsin K, is prominently reduced in the presence of curcumol. Furthermore, the molecular mechanism of action was investigated, and curcumol inhibited osteoclastogenesis by specifically impairing RANKL-induced c-Jun N-terminal kinase (JNK)/activator protein-1 (AP-1) signaling, which was further identified in rescue studies by means of anisomycin, a JNK signaling-specific activator. Taken together, these findings suggest that curcumol suppresses RANKL-induced osteoclast differentiation through the JNK/AP-1 signaling pathway, and may be useful as a therapeutic treatment for bone resorption-associated diseases.
Assuntos
Diferenciação Celular/efeitos dos fármacos , Medicamentos de Ervas Chinesas/farmacologia , Osteoclastos/efeitos dos fármacos , Sesquiterpenos/farmacologia , Animais , Anisomicina/farmacologia , Sobrevivência Celular/efeitos dos fármacos , Expressão Gênica/efeitos dos fármacos , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Camundongos , Fatores de Transcrição NFATC/metabolismo , Osteoclastos/citologia , Osteoclastos/fisiologia , Ligante RANK/farmacologia , Fator de Transcrição AP-1/metabolismoRESUMO
OBJECTIVE: To investigate the effect of Qingyi Decoction (QYD) on pancreatic gene expression profiles in rats with severe acute pancreatitis (SAP). METHODS: Totally 60 Sprague-Dawley (SD) rats were randomly divided into the sham-operation group (SO group), the SAP group, and the QYD group, 20 in each group. SAP model was replicated by the pancreatic duct retrograde injection with 4% sodium taurocholate. Rats in the QYD group was intragastrically intervened by QYD (0.75 mL/100 g) for 3 times. Pancreatic RNA expression was analyzed using Illuminate whole genome expression profiles. Changes of mRNA and protein in specific genes [heat shock proteins a8 (Hspa8) and heat shock proteins b1 (Hspb1)] were verified by real-time quantitative PCR and Western blot analysis. RESULTS: Compared with the SAP group, 575 differential genes were screened in the QYD group, including 92 up-regulated genes and 483 down-regulated genes. Gene Ontology (GO) categories indicated the genes are associated with negative regulation of transcription regulator activity, oxidoreductase activity and enzyme inhibitor activity. Effects of QYD on the SAP rats were major related to mitogen-activated protein kinase (MAPK), NOD like receptors (NLR) receptor-like signaling pathway, cell cycle, metabolic pathways, oxidoreductase activity. Protein and mRNA changes of Hspa8 and Hspb1 in microarray were verified [relative mRNA expression for Hspa8 and Hspb1 was increased by (13.24 +/- 1.22) times and (7.55 +/- 1.09) times respectively, P < 0.01]. CONCLUSION: QYD was effective in treating experimental SAP involved the MAPK and NLR signaling pathways, cell cycle, metabolic pathways, and oxide reductase activities.