RESUMO
OBJECTIVE: To study the clinical value and operation skills of nasal endoscopy-assisted bulboprostatic anastomosis in the treatment of posterior urethral stricture. METHODS: Between January 2012 and November 2014, we performed nasal endoscopy-assisted bulboprostatic anastomosis for 12 male patients with posterior urethral stricture. We recorded the operation time, blood loss, exposure of operation visual field, and success rate of anastomosis and summarized the operation skills. RESULTS: Eight of the patients experienced first-stage recovery. Two underwent a urethral dilation at 3 months postoperatively, 1 received 10 urethral dilations within 5 months after surgery, and 1 underwent internal urethrotomy after failure in urethral dilation, but all the 4 cases were cured. CONCLUSION: Nasal endoscopy can significantly improve the operation field exposure, elevate the precision, reduce the difficulty, and enhance the efficiency of bulboprostatic anastomosis in the treatment of posterior urethral stricture.
Assuntos
Anastomose Cirúrgica , Endoscopia , Estreitamento Uretral/cirurgia , Humanos , Masculino , Duração da Cirurgia , Período Pós-Operatório , Uretra/patologia , Uretra/cirurgiaRESUMO
BACKGROUND: The MYH3-associated myosinopathies comprise a spectrum of rare neuromuscular disorders mainly characterized by distal arthrogryposis with or without other features like pterygia and vertebrae fusion. CPSKF1B (contractures, pterygia, and spondylocarpotarsal fusion syndrome1B) is the only known autosomal recessiveMYH3-associated myosinopathy so far, with no more than two dozen cases being reported. MATERIALS AND METHODS: A boy with CPSKF1B was recruited and subjected to a comprehensive clinical and imaging evaluation. Genetic detection with whole-exome sequencing (WES) was performed on the patient and extended family members to identify the causative variation. A series of in silico and in vitro investigations were carried out to verify the pathogenicity of the two variants of the identified compound heterozygous variation. RESULTS: The patient exhibited moderate CPSKF1B symptoms including multiarticular contractures, webbed neck, and spondylocarpotarsal fusion. WES detected a compound heterozygous MYH3 variation consisting of two variants, namely NM_002470.4: c.3377A>G; p. (E1126G) and NM_002470.4: c.5161-2A>C. It was indicated that the NM_002470.4: c.3377A>G; p. (E1126G) variant mainly impaired the local hydrogen bond formation and impacted the TGF-B pathway, while the NM_002470.4: c.5161-2A>C variant could affect the normal splicing of pre-mRNA, resulting in the appearance of multiple abnormal transcripts. CONCLUSIONS: The findings of this study expanded the mutation spectrum of CPSKF1B, provided an important basis for the counseling of the affected family, and also laid a foundation for the functional study of MYH3 mutations.
Assuntos
Artrogripose , Túnica Conjuntiva , Contratura , Pterígio , Humanos , Masculino , Artrogripose/genética , Túnica Conjuntiva/anormalidades , Contratura/genética , FamíliaRESUMO
BACKGROUND: There is scant data on the association of the Pulsed wave-Doppler tissue imaging (PW-DTI)-derived tricuspid lateral annular peak systolic velocity (S') and poor short-term prognosis of patients with acute decompensated heart failure (ADHF). PATIENTS AND METHODS: A total number of 732 participants from the Heb-ADHF registry in China were divided into three groups according to the corresponding status of tricuspid S'. Demographic characteristics, comorbidities, physical examinations, lab tests, and medications were compared among the different groups. Different logistic regression models were utilized to gauge the relationship between S' and the risk of a composite of short-term all-cause mortality or 30-day heart failure (HF)-related rehospitalization. RESULTS: The number of composite outcome events identified in the study population was 85, with the short-term all-cause death coupled with 30-day HF readmission events reaching 23 and 62, respectively. As per the multivariable adjusted analysis, S' was inversely related to the risk of a composite outcome [<10 cm/s odds ratios (OR) 2.90, 95% confidence interval (CI):1.33-6.31; 10-11 cm/s OR 2.18, 95% CI: 1.10-4.33; p for trend = 0.006] in comparison with S' at >11 cm/s. When S' was analysed as a continuous variable, per 1 cm/s increase, the OR (95% CI) for a composite outcome was [0.87 (0.77-0.99), p = 0.028]. Area under curve (AUC) of S' for predicting outcome of ADHF was 0.631 (95%CI: 0.573-0.690, p < 0.01). Significant inverse association was also observed in left ventricular ejection fraction (LVEF) ≥40% subgroup. CONCLUSIONS: Inspite of the potential confounders, a more impaired tricuspid annular peak systolic velocity is associated with a poorer short-term prognosis of patients with ADHF.
This is the first comprehensive evaluation of tricuspid annular systolic velocity among patients with ADHF.Tricuspid annular systolic velocity could be a predictor of poor short-term prognosis in ADHF.Tricuspid annular systolic velocity should be considered in patients with ADHF at admission.
Assuntos
Ecocardiografia Doppler , Insuficiência Cardíaca , Humanos , Ecocardiografia Doppler/métodos , Prognóstico , Volume Sistólico , Função Ventricular EsquerdaRESUMO
The iron response element (IRE) is a highly conserved RNA stem loop structure. It is the binding site of iron regulatory protein (IRP). IRP binding to IRE is regulated by cellular iron. When cells are derived of iron, IRP binds IRE. If IRE is located at 5'UTR, IRP binding will inhibit translation initiation, else if IRE is at 3'UTR, IRP binding will stabilize mRNA and prevent it from degradation. So far all known IREs have C at the 1 position and G at the 5 position of the loop (C1G5 type). In vitro studies suggest that the U1A5 type IRE, which has U and A at the 1 and 5 loop position respectively, binds well to IRP. However, U1A5 type's in vivo existence is still elusive. IRE-IRP binding is involved in the regulation of iron metabolism, oxidative stress and possibly aging. Here we use an improved computation method performing a comprehensive search of IRE in human and mouse genes. We try to catalog potential human and mouse IRE containing genes, at the same time identify potential U1A5 IREs.
Assuntos
Regiões 3' não Traduzidas/genética , Regiões 5' não Traduzidas/genética , Ferro/farmacologia , Elementos de Resposta/genética , Software , Regiões 3' não Traduzidas/química , Regiões 5' não Traduzidas/química , Animais , Sequência de Bases , Sítios de Ligação/genética , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Ferro/metabolismo , Proteínas de Ligação ao Ferro/metabolismo , Camundongos , Conformação de Ácido Nucleico , Elementos de Resposta/efeitos dos fármacosRESUMO
This paper reports the construction of the database for Genomic Polymorphism of Chinese Ethnic Groups (GPCEG). GPCEG contains denomination and basic information of Chinese 56 ethnic groups, with introduction of their in geographic distribution, population quantity, spoken and written language, religious belief and physical characteristics. GPCEG collects the data of genomic polymorphism, cell lines, reference and links of other international related databases. The visualization, query and update system were also available. GPCEG laid the foundations of establishing a national database with Chinese characteristics.
Assuntos
Bases de Dados de Ácidos Nucleicos , Genoma Humano , Polimorfismo Genético , Linhagem Celular , China , Frequência do Gene , Marcadores Genéticos , Haplótipos , HumanosRESUMO
Porcine reproductive and respiratory syndrome (PRRS), caused by porcine reproductive and respiratory syndrome virus (PRRSV), is an acute infectious disease. The prevalence of PRRS has made swine industry suffered huge financial losses. Matrine, a natural compound, has been demonstrated to possess anti-PRRSV activity in Marc-145 cells. However, the underlying molecular mechanisms were still unknown. The main objective of our study was to discuss the effect of Matrine on PRRSV N protein expression and PRRSV induced apoptosis. Indirect immunofluorescence assay (IFA) and Western blot were used to assess the effect of Matrine on N protein expression. Apoptosis was analyzed by fluorescence staining. In addition, the effect of Matrine on caspase-3 activation was investigated by Western blot. Indirect immunofluorescence assay and Western blot analysis demonstrated that Matrine could inhibit N protein expression in Marc-145 cells. And Matrine was found to be able to impair PRRSV-induced apoptosis by inhibiting caspase-3 activation.
Assuntos
Alcaloides/farmacologia , Apoptose/imunologia , Síndrome Respiratória e Reprodutiva Suína/tratamento farmacológico , Vírus da Síndrome Respiratória e Reprodutiva Suína/imunologia , Quinolizinas/farmacologia , Alcaloides/uso terapêutico , Animais , Apoptose/efeitos dos fármacos , Western Blotting/veterinária , Linhagem Celular , Chlorocebus aethiops , Microscopia de Fluorescência/veterinária , Proteínas do Nucleocapsídeo/imunologia , Síndrome Respiratória e Reprodutiva Suína/imunologia , Quinolizinas/uso terapêutico , Suínos , MatrinasRESUMO
BACKGROUND: Porcine reproductive and respiratory syndrome virus (PRRSV) has caused large economic losses in the swine industry. Currently, there is no effective way to prevent PRRSV infection. In this study, we investigated the inhibitory effect of dipotassium glycyrrhetate (DG), a derivative of glycyrrhetinic acid, on PRRSV infection ability. METHODS: The cytotoxicity of DG was measured by MTT assay, and the effects of DG on PRRSV N gene/protein were investigated using real-time PCR, western blot and immunofluorescence assay. In addition, the effect of DG on cell apoptosis was analysed by fluorescence staining. RESULTS: Our results indicated that DG could effectively inhibit virus replication and N gene expression in MARC-145 cells infected with PRRSV. When the infected cells received DG, the numbers of apoptotic cells were decreased, and the cleaved caspase-3 contents were decreased dramatically. CONCLUSIONS: Our study demonstrates that DG could effectively inhibit the PRRS virus via multiple pathways including inhibition of virus replication and N gene expression and reduction of apoptotic cells. DG can serve as a potential chemical for PRRSV prevention and control.
Assuntos
Apoptose/efeitos dos fármacos , Ácido Glicirrízico/farmacologia , Proteínas do Nucleocapsídeo/biossíntese , Síndrome Respiratória e Reprodutiva Suína/tratamento farmacológico , Replicação Viral/efeitos dos fármacos , Animais , Anti-Inflamatórios/farmacologia , Antivirais/farmacologia , Caspase 3/metabolismo , Linhagem Celular , Chlorocebus aethiops , Expressão Gênica/efeitos dos fármacos , Síndrome Respiratória e Reprodutiva Suína/prevenção & controle , Vírus da Síndrome Respiratória e Reprodutiva Suína/efeitos dos fármacos , SuínosRESUMO
Chickens experimentally infected with Marek's disease virus J-1 strain were used to evaluate the anti-Marek's disease virus (MDV) activity of sodium tanshinone IIA sulfonate (STS) in vivo. Chickens in same group were kept in one pen and control group chickens were housed in negative pressure isolator. Chickens were treated with different dose of STS or ABOB for 21 consecutive days. Peripheral T lymphocyte proliferation, expression level of IFN-γ and IL-10 in serum, and MDV load in spleen were determined. The results showed that the treatments with STS and ABOB could significantly increase stimulating index (SI) of peripheral T lymphocytes while the SI is dropping due to the MDV infection, SI of chickens in STS prevention groups were significantly higher than that in STS treatment group and ABOB group (P<0.05 or P<0.01); IFN-γ and IL-10 level of chickens in STS groups were higher than that in other groups (P<0.05 or P<0.01). The results of qPCR demonstrated that STS could inhibit the virus replication in spleen of chickens infected with MDV. These findings indicated that STS can be potentially applied as an anti-MDV drug and set a solid basis for further investigating the antiviral mechanisms of STS.
Assuntos
Antivirais/uso terapêutico , Doença de Marek/tratamento farmacológico , Fenantrenos/uso terapêutico , Doenças das Aves Domésticas/tratamento farmacológico , Animais , Antivirais/farmacologia , Proteínas Aviárias/sangue , Proliferação de Células/efeitos dos fármacos , Galinhas , Avaliação Pré-Clínica de Medicamentos , Interferon gama/sangue , Interleucina-10/sangue , Doença de Marek/sangue , Doença de Marek/virologia , Fenantrenos/farmacologia , Doenças das Aves Domésticas/sangue , Doenças das Aves Domésticas/virologia , Baço/virologia , Linfócitos T/efeitos dos fármacos , Linfócitos T/fisiologia , Linfócitos T/virologia , Carga ViralAssuntos
Analgésicos Opioides/farmacologia , Inibidor da Ligação a Diazepam/farmacologia , Diazepam/farmacologia , Morfina/farmacologia , Receptores de GABA-A/metabolismo , Analgésicos Opioides/química , Animais , Bovinos , Diazepam/química , Inibidor da Ligação a Diazepam/síntese química , Inibidor da Ligação a Diazepam/química , Relação Dose-Resposta a Droga , Camundongos , Morfina/química , SuínosRESUMO
AIM: Ca2+ release from the endoplasmic reticulum (ER) is an integral component of neuronal Ca2+ signaling. The present study is to investigate properties of local Ca2+ release events in superior cervical ganglion (SCG) neurons. METHODS: Primary cultured SCG neurons were prepared from neonatal rats (P3-P7). Low concentration of caffeine was used to induce Ca2+ release from the ER Ca2+ store, and intracellular Ca2+ was recorded by high-resolution line scan confocal imaging and the Ca2+ indicator Fluo-4. RESULTS: Two populations of local Ca2+ release events with distinct temporal characteristics were evoked by 1.5 mmol/L caffeine near the surface membrane in the soma and the neurites of SCG neurons. Brief events similar to classic Ca2+ sparks lasted a few hundreds of milliseconds, whereas long-lasting events displayed duration up to tens of seconds. Typical somatic and neurite sparks were of 0.3- and 0.52-fold increase in local Fluo-4 fluorescence, respectively. Typical Ca2+ glows were brighter (deltaF/F0 approximately 0.6), but were highly confined in space. The half maximum of full duration of neurite sparks was much longer than those in the soma (685 vs 381 ms). CONCLUSION: Co-existence of Ca2+ sparks and Ca2+ glows in SCG neurons indicates distinctive local regulation of Ca2+ release kinetics. The local Ca2+ signals of variable, site-specific temporal length may bear important implications in encoding a 'memory' of the trigger signal.
Assuntos
Sinalização do Cálcio/efeitos dos fármacos , Cálcio/metabolismo , Retículo Endoplasmático/metabolismo , Gânglio Cervical Superior/metabolismo , Animais , Animais Recém-Nascidos , Cafeína/antagonistas & inibidores , Células Cultivadas , Neurônios/metabolismo , Ratos , Gânglio Cervical Superior/citologia , Tapsigargina/farmacologiaRESUMO
To elucidate the temperature dependence and underlying thermodynamic determinants of the elementary Ca2+ release from the sarcoplasmic reticulum, we characterized Ca2+ sparks originating from ryanodine receptors (RyRs) in rat cardiomyocytes over a wide range of temperature. From 35 degrees C to 10 degrees C, the normalized fluo-3 fluorescence of Ca2+ sparks decreased monotonically, but the Delta[Ca2+]i were relatively unchanged due to increased resting [Ca2+]i. The time-to-peak of Ca2+ sparks, which represents the RyR Ca2+ release duration, was prolonged by 37% from 35 degrees C to 10 degrees C. An Arrhenius plot of the data identified a jump of apparent activation energy from 5.2 to 14.6 kJ/mol at 24.8 degrees C, which presumably reflects a transition of sarcoplasmic reticulum lipids. Thermodynamic analysis of the decay kinetics showed that active transport plays little role in early recovery but a significant role in late recovery of local Ca2+ concentration. These results provided a basis for quantitative interpretation of intracellular Ca2+ signaling under various thermal conditions. The relative temperature insensitivity above the transitional 25 degrees C led to the notion that Ca2+ sparks measured at a "warm room" temperature are basically acceptable in elucidating mammalian heart function.
Assuntos
Sinalização do Cálcio/fisiologia , Cálcio/metabolismo , Modelos Biológicos , Miócitos Cardíacos/fisiologia , Canal de Liberação de Cálcio do Receptor de Rianodina/metabolismo , Temperatura , Animais , Células Cultivadas , Simulação por Computador , Cinética , Ratos , Ratos Sprague-Dawley , TermodinâmicaRESUMO
We used voltage-sensitive dye imaging to visualize the distribution of initiation sites of the spontaneous interictal-like spikes (sISs) in rat neocortex, in vivo, induced by bicuculline or picrotoxin over the exposed cortex. The initiation site was small (approximately 200 microm diam). On average each initiation site initiated 2.0 +/- 0.8 sISs (9 animals, 499 sISs, 251 sites). This is significantly different from that in neocortical slices, where each initiation site initiated 30-100 sISs. The initiation sites were not randomly distributed. The distance between two consecutive sites tended to be either <800 or >1200 microm, suggesting a temporal "suppression annulus" surrounding each initiation site. Within the annulus, the likelihood for initiating the next sIS was reduced. Suppression annulus did not have a noticeable change in the presence of GABA(b) antagonist, suggesting it did not depend on the GABA(b) inhibition. We also applied bicuculline locally to a spot of 800 x 800 microm(2) for approximately 45 min. During this period approximately 1000 sISs occurred within the spot. Bicuculline or picrotoxin was then applied to the entire craniotomy window. The pretreatment created an obvious cluster of initiation sites. Around this cluster, the suppression annulus became obvious in individual animals. Our results suggest that, in disinhibited cortex, epileptiform events were initiated from small sites. The initiation sites may cluster in an area with increased local activity. Surrounding each initiation site there may be a temporal suppression annulus.
Assuntos
Epilepsia/fisiopatologia , Potenciais Evocados/fisiologia , Neocórtex/fisiologia , Animais , Bicuculina/farmacologia , Potenciais Evocados/efeitos dos fármacos , Antagonistas de Receptores de GABA-B , Masculino , Neocórtex/efeitos dos fármacos , Picrotoxina/farmacologia , Ratos , Ratos Sprague-Dawley , Receptores de GABA-B/fisiologiaRESUMO
AIM: To study the effect of puerarin (Pue) on Na+ channel in rat ventricular myocytes. METHODS: Whole-cell patch-clamp technique was applied on isolated cardiomyocytes from rats. RESULTS: Pue inhibited cardiac INa in a positive rate-dependent and dose-dependent manner, with an IC(50) of 349 micromol/L. The kinetics of blockage of cardiac sodium channel by Pue resembled the ClassIa/Ic of antiarrhythmic agents. Pue 300 micromol/L did not alter the shape of the I-V curve of INa, but markedly shifted the steady-state inactivation curve of INa towards more negative potential by 15.9 mV, and postponed the recovery of INa inactivation state from (21.9+/-1.6) ms to (54.4+/-3.4) ms (P<0.01). It demonstrated that the steady state of inactivation was affected by Pue significantly. CONCLUSION: Pue protected ventricular myocytes against cardiac damage and arrhythmias by inhibiting recovery from inactivation of cardiac Na+ channels.
Assuntos
Isoflavonas/farmacologia , Miócitos Cardíacos/efeitos dos fármacos , Bloqueadores dos Canais de Sódio/farmacologia , Canais de Sódio/efeitos dos fármacos , Animais , Separação Celular , Feminino , Ventrículos do Coração/citologia , Masculino , Miocárdio/citologia , Miócitos Cardíacos/metabolismo , Ratos , Ratos Sprague-DawleyRESUMO
AIM: To study the blockade of paeoniflorin (Pae) on I(Na) in the acutely isolated hippocampus neurons of mice. METHODS: The whole-cell patch clamp technique was used. RESULTS: Pae inhibited I(Na) in frequency-dependent and concentration-dependent manners, with an IC50 of 271 micromol/L. Pae 0.3 mmol/L shifted the activation potential of the maximal I(Na) from -40 mV to -30 mV, shifted the steady-state activation and inactivation curves toward more positive and negative potentials by 10.8 mV, and 18.2 mV, respectively, and postponed the recovery of I(Na) inactivation state from (4.2+/-0.7) ms to (9.8+/-1.2) ms. CONCLUSION: Pae inhibited I(Na) in mouse hippocampus neurons.
Assuntos
Benzoatos/farmacologia , Hidrocarbonetos Aromáticos com Pontes/farmacologia , Glucosídeos/farmacologia , Hipocampo/citologia , Neurônios/efeitos dos fármacos , Bloqueadores dos Canais de Sódio/farmacologia , Canais de Sódio/metabolismo , Animais , Benzoatos/isolamento & purificação , Hidrocarbonetos Aromáticos com Pontes/isolamento & purificação , Glucosídeos/isolamento & purificação , Potenciais da Membrana/efeitos dos fármacos , Camundongos , Monoterpenos , Neurônios/fisiologia , Fármacos Neuroprotetores/farmacologia , Paeonia/química , Plantas Medicinais/químicaRESUMO
AIM: To assess the blockade by CPU 86017 on the L-type calcium channels in the myocardium and on the Ca(2+)-related contractions of vascular smooth muscle. METHODS: The whole-cell patch-clamp was applied to investigate the blocking effect of CPU 86017 on the L-type calcium current in isolated guinea pig myocytes and contractions by KCl or phenylephrine (Phe) of the isolated rat tail arteries were measured. RESULTS: Suppression of the L-type current of the isolated myocytes by CPU 86017 was moderate, in time- and concentration-dependent manner and with no influence on the activation and inactivation curves. The IC(50) was 11.5 micromol/L. Suppressive effect of CPU 86017 on vaso-contractions induced by KCl 100 mmol/L, phenylephrine 1 micromol/L in KH solution (phase 1), Ca(2+) free KH solution ( phase 2), and by addition of CaCl(2) into Ca(2+)-free KH solution (phase 3) were observed. The IC(50) to suppress vaso-contractions by calcium entry via the receptor operated channel (ROC) and voltage-dependent channel (VDC) was 0.324 micromol/L and 16.3 micromol/L, respectively. The relative potency of CPU 86017 to suppress vascular tone by Ca(2+) entry through ROC and VDC is 1/187 of prazosin and 1/37 of verapamil, respectively. CONCLUSION: The blocking effects of CPU 86017 on the L-type calcium channel of myocardium and vessel are moderate and non-selective. CPU 86017 is approximately 50 times more potent in inhibiting ROC than VDC.
Assuntos
Berberina/análogos & derivados , Berberina/farmacologia , Canais de Cálcio Tipo L/efeitos dos fármacos , Contração Muscular/efeitos dos fármacos , Músculo Liso Vascular/efeitos dos fármacos , Miocárdio/metabolismo , Animais , Berberina/administração & dosagem , Bloqueadores dos Canais de Cálcio/farmacologia , Células Cultivadas , Relação Dose-Resposta a Droga , Feminino , Cobaias , Masculino , Miocárdio/citologia , Técnicas de Patch-Clamp , Ratos , Ratos Sprague-Dawley , Cauda/irrigação sanguíneaRESUMO
Abnormal mechanical load, as seen in hypertension, is found to induce heart cell apoptosis, yet the signaling link between cell stretch and apoptotic pathways is not known. Using an in vitro stretch model mimicking diastolic pressure stress, here we show that Ca(2+) signaling participates essentially in the early stage of stretch-induced apoptosis. In neonatal rat cardiomyocytes, the moderate 20% stretch resulted in tonic elevation of intracellular free Ca(2+) ([Ca(2+)](i)). Buffering [Ca(2+)](i) by EGTA-AM, suppressing ryanodine-sensitive Ca(2+) release, and blocking L-type Ca(2+) channels all prevented the stretch-induced apoptosis as assessed by phosphatidylserine exposure and nuclear fragmentation. Notably, Ca(2+) suppression also prevented known stretch-activated apoptotic events, including caspase-3/-9 activation, mitochondrial membrane potential corruption, and reactive oxygen species production, suggesting that Ca(2+) signaling is the upstream of these events. Since [Ca(2+)](i) did not change without activating mechanosensitive Ca(2+) entry, we conclude that stretch-induced Ca(2+) entry, via the Ca(2+)-induced Ca(2+) release mechanism, plays an important role in initiating apoptotic signaling during mechanical stress.