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PURPOSE: To explore the genetic etiology of deafness in a dominant family with late-onset, progressive, nonsyndromic hearing loss. METHODS: Genome-wide linkage analysis was performed for 21 family members. Candidate pathogenic variants were identified by whole-exome sequencing of selected family members and confirmed by Sanger sequencing of all family members. Cochlear expression of Dmxl2 was investigated by reverse-transcription polymerase chain reaction (RT-PCR) and immunostaining of the organ of Corti from mice. RESULTS: The causative gene was mapped to a 9.68-Mb candidate region on chromosome 15q21.2 (maximum logarithm of the odds score = 4.03) that contained no previously described deafness genes. Whole-exome sequencing identified heterozygous c.7250G>A (p.Arg2417His) in DMXL2 as the only candidate pathogenic variant segregating the hearing loss. In mouse cochlea, expression of DMXL2 was restricted to the hair cells and the spiral ganglion neurons. CONCLUSION: Our data indicated that the p.Arg2417His variant in DMXL2 is associated with dominant, nonsyndromic hearing loss and suggested an important role of DMXL2 in inner ear function.Genet Med advance online publication 22 September 2016.
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Proteínas Adaptadoras de Transdução de Sinal/genética , Surdez/genética , Mutação de Sentido Incorreto , Proteínas do Tecido Nervoso/genética , Órgão Espiral/metabolismo , Proteínas Adaptadoras de Transdução de Sinal/metabolismo , Idade de Início , Animais , China/etnologia , Surdez/metabolismo , Feminino , Estudos de Associação Genética , Ligação Genética , Predisposição Genética para Doença , Humanos , Masculino , Camundongos , Proteínas do Tecido Nervoso/metabolismo , Linhagem , Mapeamento Físico do Cromossomo , Sequenciamento do ExomaRESUMO
Metallothionein-2 (MT-2) was originally discovered as a mediator of zinc homeostasis and cadmium detoxification. However, MT-2 has recently received increased attention because altered expression of MT-2 is closely related to various diseases such as asthma and cancers. Several pharmacological strategies have been developed to inhibit or modify MT-2, revealing its potential as drug target in diseases. Therefore, a better understanding of the mechanisms of MT-2 action is warranted to improve drug development for potential clinical applications. In this review, we highlight recent advances in determining the protein structure, regulation, binding partners, and new functions of MT-2 in inflammatory diseases and cancers.
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Neoplasias , Zinco , Humanos , Zinco/metabolismo , Metalotioneína/metabolismo , Neoplasias/tratamento farmacológico , Cádmio/metabolismoRESUMO
Asthma is a complex and heterogeneous respiratory disease that causes serious social and economic burdens. Current drugs such as ß2-agonists cannot fully control asthma. Our previous study found that Transgelin-2 is a potential target for treating asthmatic pulmonary resistance. Herein, we discovered a zolinium compound, TSG1180, that showed a strong interaction with Transgelin-2. The equilibrium dissociation constants (KD) of TSG1180 to Transgelin-2 were determined to be 5.363 × 10-6 and 9.81 × 10-6 M by surface plasmon resonance (SPR) and isothermal titration calorimetry (ITC). Cellular thermal shift assay (CETSA) results showed that the thermal stability of Transgelin-2 increased after coincubation of TSG1180 with lysates of airway smooth muscle cells (ASMCs). Molecular docking showed that Arg39 may be the key residue for the binding. Then, the SPR result showed that the binding affinity of TSG1180 to Transgelin-2 mutant (R39E) was decreased by 1.69-fold. Real time cell analysis (RTCA) showed that TSG1180 treatment could relax ASMCs by 19 % (P < 0.05). Once Transgelin-2 was inhibited, TSG1180 cannot induce a relaxation effect, suggesting that the relaxation effect was specifically mediated by Transgelin-2. In vivo study showed TSG1180 effectively reduced pulmonary resistance by 64 % in methacholine-induced mice model (P < 0.05). Furthermore, the phosphorylation of Ezrin at T567 was increased by 8.06-fold, the phosphorylation of ROCK at Y722 was reduced by 38 % and the phosphorylation of RhoA at S188 was increased by 52 % after TSG1180 treatment. These results suggested that TSG1180 could be a Transgelin-2 agonist for further optimization and development as an anti-asthma drug.
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Asma , Camundongos , Animais , Simulação de Acoplamento Molecular , Asma/tratamento farmacológico , Asma/metabolismo , Pulmão , Proteínas dos Microfilamentos/metabolismo , Miócitos de Músculo Liso/metabolismoRESUMO
Airway hyperresponsiveness (AHR) is one of the most important features of asthma. Our previous study showed that inhaled transgelin-2 agonist, TSG12, effectively reduced pulmonary resistance in a mouse model of asthma in a dose-dependent manner. However, the optimal administration time of TSG12 to reduce AHR and the pharmacological effects are still unclear. In this study, the effects of TSG12 inhalation before and during AHR occurrence were examined. The results showed that the pulmonary resistance was reduced by 57% and the dynamic compliance was increased by 46% in the TSG12 Mch group (atomize TSG12 10 min before methacholine, p < 0.05 vs. model). The pulmonary resistance was reduced by 61% and the dynamic compliance was increased by 47% in the TSG12 + Mch group (atomize TSG12 and methacholine together, p < 0.05 vs. model). Quantitative real-time PCR showed that the gene expression levels of transgelin-2, myosin phosphatase target subunit-1, and myosin light chain were up-regulated by 6.4-, 1.9-, and 2.8-fold, respectively, in the TSG12 Mch group. The gene expression levels of transgelin-2, myosin phosphatase target subunit-1, and myosin light chain were up-regulated by 3.2-, 1.4-, and 1.9-fold, respectively, in the TSG12 + Mch group. The results suggested that TSG12 effectively reduces pulmonary resistance when TSG12 inhalation occurred both before and during AHR occurrence. Gene expression levels of transgelin-2 and myosin light chain were significantly up-regulated when TSG12 inhalation occurred before AHR occurrence. This study may provide a basis for the administration time of TSG12 for asthma treatment in the future.
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OBJECTIVE: Study on the application of WAIS-RC short forms and adult intelligence disability scale in mental impairment assessment. METHODS: Mental impairment assessment cases between July 2009 and March 2011 in judicial appraisal institute of Taizhou University were collected. Assessment results obtained with the WAIS-RC short forms and adult intelligence disability scale were compared with the experts assessing conclusions and analyzed using SPSS 11.5 software. RESULTS: Assessment results with the two scales did not fully comply with the expert's conclusions, with reliability coefficient were 0.785 and 0.940 respectively, correlation coefficient were 0.850 and 0.922 respectively. CONCLUSION: The intelligence assessment was influenced by many factors. When the appraised individuals had nerve dysfunction and mild intelligence disability or mental disorders, the two scales should be used together. When the appraised individuals had moderate intelligence disability or mental disorders, adult intelligence disability scale had advantage.
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Deficiência Intelectual/diagnóstico , Inteligência , Escalas de Wechsler , Acidentes de Trânsito , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Lesões Encefálicas/complicações , Avaliação da Deficiência , Prova Pericial , Feminino , Psiquiatria Legal , Humanos , Deficiência Intelectual/psicologia , Masculino , Transtornos Mentais/diagnóstico , Transtornos Mentais/psicologia , Pessoa de Meia-Idade , Reprodutibilidade dos Testes , Índice de Gravidade de Doença , Escalas de Wechsler/normas , Escalas de Wechsler/estatística & dados numéricos , Adulto JovemRESUMO
Strengthening the compliance is conductive to the quality improvement of clinical trial of acupuncture and moxibustion. In terms to planning behavior and influencing factors of loyalty, the questionnaire was conducted among 200 participants on the compliance of clinical trial on knee osteoarthritis treated with acupuncture. The results showed that the subjective norms and perceptual behavior control of subjects affected their compliance in the trial. Medical service compensation became the primary factor of the subjects' loyalty, which further affected their compliance in the trial. It is suggested that the compliance should be managed according to the characteristics of different clinical researches, and the feasible medical service compensation scheme should be designed in advance by actively focusing on the subjects' features.
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Terapia por Acupuntura , Moxibustão , Osteoartrite do Joelho , Humanos , Osteoartrite do Joelho/terapia , Inquéritos e QuestionáriosRESUMO
OBJECTIVE: To evaluate recovering consciousness effect of electroacupuncture (EA) on patients after traumatic brain injury (TBI) surgery. METHODS: A total of 100 patients with traumatic coma were randomly divided into an observation group and a control group, 50 cases in each group. The control group was mainly treated with awakening drugs and neurotrophic drugs; on the basis of treatment in the control group, the observation group was treated with EA at Neiguan (PC 6) and Shuigou (GV 26) with disperse-dense wave, 2 Hz/100 Hz in frequency, 0.1-5 mA in intensity. After 30 min of EA, the needles were stayed 60 min. The treatment was performed once a day for 14 consecutive days. The changes in Glasgow coma score (GCS) was observed in the two groups before treatment and after 7, 14 days of treatment; and the two groups were followed up for 3 months after treatment to evaluate the Glasgow outcome scale (GOS) and Barthel index (BI) scores. RESULTS: After 7, 14 days of treatment, the GCS scores of the two groups were higher than those before treatment (P<0.05), and the increase degree in the observation group was significantly larger than that in the control group (P<0.05). At 3 months of follow-up, the GOS and BI scores of the observation group were better than those of the control group (P<0.05). CONCLUSION: Early electroacupuncture intervention can effectively promote the recovery of consciousness after traumatic brain injury surgery, and has a curative long-term effect.
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Lesões Encefálicas Traumáticas/cirurgia , Lesões Encefálicas Traumáticas/terapia , Estado de Consciência , Eletroacupuntura , Pontos de Acupuntura , HumanosRESUMO
We aimed to explore the role of SIRT1 in apoptosis in human kidney proximal tubule epithelial (HK-2) cells, and to determine whether resveratrol (RSV, a SIRT1 activator) could ameliorate apoptosis in rats with streptozotocin-induced diabetes mellitus (DM) and/or in high glucose (HG, 30mM) - stimulated HK-2 cells. Rats were distributed randomly into three groups: 1) control group, 2) DM group, and 3) DM with RSV group (DM+RSV; rats treated with 30mg/kg/d of RSV for 16 weeks). The physical, biochemical, and morphological parameters were then examined. Additionally, the deacetylase activity of SIRT1, and the expression levels of SIRT1 and of representative apoptosis markers, such as p53, acetylated p53, cleaved caspase-3, caspase-9, and cleaved PARP, were measured. HK-2 cells were stimulated by HG for different lengths of time to study the effect of HG on apoptosis. HK-2 cells were treated with or without RSV (25µM) to investigate if RSV has a protective effect on HG-induced apoptosis. A gene-specific small interfering RNA against SIRT1 was used to study the role of SIRT1 in apoptosis. More apoptosis was found in the DM rats than in the control rats. Similarly, the expression levels of cleaved caspase-3, cleaved PARP, and acetylated p53 were significantly higher, and the level of SIRT1 was significantly lower, in the HK-2 cells that were cultured under HG conditions than those in the HK-2 cells that were cultured under low glucose (5.5mM) conditions. Notably, treatment with RSV lessened the HG-induced changes in the levels of apoptosis indicators, and this inhibition of HG-induced apoptosis in HK-2 cells by RSV treatment was abolished by SIRT1 silencing. Our study showed that hyperglycemia contributes to apoptosis in rat kidney and HK-2 cells. SIRT1 activation by RSV can reduce urinary albumin excretion and proximal tubule epithelial apoptosis both in vitro and in vivo. Based on our study, SIRT1/p53 axis played an important role in the hyperglycemia induced apoptosis. These findings indicated that the increased expression of SIRT1, mediated by RSV, is a possible mechanism by which RSV prevents renal tubular injury in diabetic nephropathy (DN). So RSV has great clinical significance and could provide the basis for the new way to effective treatment to contain the morbidity and mortality associated with DN.
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Apoptose/efeitos dos fármacos , Hiperglicemia/complicações , Túbulos Renais Proximais/patologia , Sirtuína 1/metabolismo , Estilbenos/farmacologia , Animais , Células Cultivadas , Diabetes Mellitus Experimental/complicações , Diabetes Mellitus Experimental/tratamento farmacológico , Diabetes Mellitus Experimental/patologia , Inativação Gênica/efeitos dos fármacos , Glucose/toxicidade , Humanos , Túbulos Renais Proximais/efeitos dos fármacos , Ratos Wistar , Resveratrol , EstreptozocinaRESUMO
OBJECTIVE: To investigate the mechanism of survivin, p53 and Ki-67 on Hep-2 human laryngeal cancer endothelial cell proliferation and invasion. METHODS: Laryngeal squamous cell carcinoma and paracancerous normal tissues were collected, total RNA was extracted from tissues, survivin, p53 and Ki-67 gene mRNA expression levels in laryngeal cancer and the adjacent tissues were detected by Real-time PCR. Human laryngeal cancer Hep-2 epithelial cells were selected, survivin gene was overexpressed, and cell proliferation was detected by MTT. p53 and Ki-67 gene expression changes in overexpressed survivin gene were detected by Western blot. Changes in Hep-2 cell invasive ability were studied when survivin was overexpressed as detected by Transwell invasion assay. RESULTS: In the adjacent tissues, survivin, p53 and Ki-67 gene relative expression levels were 1.72 ± 0.9, 13.7 ± 5.7 and 5.7 ± 1.3, respectively; while in cancer tissues, gene relative expression levels were 53.7 ± 8.3, 66.7 ± 5.2 and 61.0 ± 3.1, respectively, which was significantly increased. As detected by MTT, relative cell survival rate within 12 h of survivin overexpression were: load control group (88.5 ± 1.6)%; overexpressed group (90.3 ± 1.9)%. Transwell invasion assay results indicated that overexpressed survivin could significantly increase the relative survival rate of cells. CONCLUSIONS: Expressions of p53, Ki67 and survivin are increased in cancer; and there is a positive correlation between survivin, p53 and Ki67 expressions in laryngeal carcinoma.
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Diabetic nephropathy is a major cause of morbidity and mortality in diabetic patients. Effective therapies to prevent the development of this disease are required. Berberine (BBR) has several preventive effects on diabetes and its complications. However, the molecular mechanism of BBR on kidney function in diabetes is not well defined. Here, we reported that activation of AMP-activated protein kinase (AMPK) is required for BBR-induced improvement of kidney function in vivo. AMPK phosphorylation and activity, productions of reactive oxygen species (ROS), kidney function including serum blood urea nitrogen (BUN), creatinine clearance (Ccr), and urinary protein excretion, morphology of glomerulus were determined in vitro or in vivo. Exposure of cultured human glomerulus mesangial cells (HGMCs) to BBR time- or dose-dependently activates AMPK by increasing the thr172 phosphorylation and its activities. Inhibition of LKB1 by siRNA or mutant abolished BBR-induced AMPK activation. Incubation of cells with high glucose (HG, 30 mM) markedly induced the oxidative stress of HGMCs, which were abolished by 5-aminoimidazole-4-carboxamide ribonucleoside, AMPK gene overexpression or BBR. Importantly, the effects induced by BBR were bypassed by AMPK siRNA transfection in HG-treated HGMCs. In animal studies, streptozotocin-induced hyperglycemia dramatically promoted glomerulosclerosis and impaired kidney function by increasing serum BUN, urinary protein excretion, and decreasing Ccr, as well as increased oxidative stress. Administration of BBR remarkably improved kidney function in wildtype mice but not in AMPKα2-deficient mice. We conclude that AMPK activation is required for BBR to improve kidney function in diabetic mice.
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Proteínas Quinases Ativadas por AMP/metabolismo , Berberina/administração & dosagem , Diabetes Mellitus Experimental/metabolismo , Nefropatias Diabéticas/metabolismo , Animais , Berberina/farmacologia , Linhagem Celular , Diabetes Mellitus Experimental/tratamento farmacológico , Nefropatias Diabéticas/tratamento farmacológico , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Testes de Função Renal , Células Mesangiais/metabolismo , Camundongos , Estresse Oxidativo/efeitos dos fármacos , Fosforilação , EstreptozocinaRESUMO
The purpose of this study is to investigate the mRNA and protein expression levels of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) in pressure ulcers, and to elucidate the molecular mechanism by which VEGF and bFGF are involved in pressure ulcer formation. A rat model of ischemia-reperfusion pressure ulcer was established by magnetic disk circulating compression method. Real-time fluorescence quantitative PCR and Western blot assays were conducted to detect the mRNA and protein expression of VEGF and bFGF in the tissues of rat I-, II-, and III-degree pressure ulcers, the surrounding tissues, and normal skin. Our study confirmed that the mRNA and protein expression levels of VEGF and bFGF in the tissues of rat I-degree pressure ulcer were significantly higher than that in the II- and III-degree pressure ulcer tissues (P < 0.05). The expression of VEGF and bFGF in the tissues surrounding I- and II-degree pressure ulcers were higher than the rats with normal skin. The expression of VEGF and bFGF in the tissues of rat III-degree pressure ulcer was lower than that in the surrounding tissues and normal skin (P < 0.05). There was a significant positive correlation between change in the VEGF and bFGF. The results showed that with an increase in the degree of pressure ulcers, the expression of VEGF and bFGF in pressure ulcers tissue are decreased. This leads to a reduction in angiogenesis and may be a crucial factor in the formation of pressure ulcers.