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BACKGROUND: Members of the nucleotide-binding oligomerization domain, leucine rich repeat and pyrin domain containing (NLRP) family regulate various physiological and pathological processes. However, none have been shown to regulate actin cap formation or spindle translocation during the asymmetric division of oocyte meiosis I. NLRP4E has been reported as a candidate protein in female fertility, but its function is unknown. METHODS: Immunofluorescence, reverse transcription polymerase chain reaction (RT-PCR), and western blotting were employed to examine the localization and expression levels of NLRP4E and related proteins in mouse oocytes. small interfering RNA (siRNA) and antibody transfection were used to knock down NLRP4E and other proteins. Immunoprecipitation (IP)-mass spectrometry was used to identify the potential proteins interacting with NLRP4E. Coimmunoprecipitation (Co-IP) was used to verify the protein interactions. Wild type (WT) or mutant NLRP4E messenger RNA (mRNA) was injected into oocytes for rescue experiments. In vitro phosphorylation was employed to examine the activation of steroid receptor coactivator (SRC) by NLRP4E. RESULTS: NLRP4E was more predominant within oocytes compared with other NLRP4 members. NLRP4E knockdown significantly inhibited actin cap formation and spindle translocation toward the cap region, resulting in the failure of polar body extrusion at the end of meiosis I. Mechanistically, GRIN1, and GANO1 activated NLRP4E by phosphorylation at Ser429 and Thr430; p-NLRP4E is translocated and is accumulated in the actin cap region during spindle translocation. Next, we found that p-NLRP4E directly phosphorylated SRC at Tyr418, while p-SRC negatively regulated p-CDC42-S71, an inactive form of CDC42 that promotes actin cap formation and spindle translocation in the GTP-bound form. CONCLUSIONS: NLRP4E activated by GRIN1 and GANO1 regulates actin cap formation and spindle translocation toward the cap region through upregulation of p-SRC-Tyr418 and downregulation of p-CDC42-S71 during meiosis I.
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Actinas , Meiosis , Oocitos , Proteína de Unión al GTP cdc42 , Animales , Oocitos/metabolismo , Ratones , Femenino , Actinas/metabolismo , Actinas/genética , Proteína de Unión al GTP cdc42/metabolismo , Proteína de Unión al GTP cdc42/genética , Fosforilación , Huso Acromático/metabolismoRESUMEN
Uterine dysplasia is a common cause of infertility. Traditional Chinese medicine has unique advantages in the treatment of this disease. This paper introduces a case of infertility caused by uterine dysplasia treated by Professor MA Kun who adopted the therapy of tonifying kidney and activating blood, aiming to summarize the theoretical foundation and formula principles of Professor MA Kun in the clinical treatment of this disease. The kidney stores essence and governs reproduction. Kidney deficiency is the root cause of infertility. The deficiencies in kidney Qi, Yin, and Yang can result in blood stasis to obstruct the uterus, leading to insufficient source for essence and aggravating kidney deficiency. Kidney deficiency and blood stasis affect each other and form a vicious cycle, resulting in uterine dysplasia due to insufficient nutrition and difficult pregnancy. Therefore, Professor MA Kun believes that kidney deficiency and blood stasis is the key pathogenesis of infertility caused by uterine dysplasia and proposes the treatment principle of tonifying kidney and activating blood. Sufficient essence and Qi in the kidney can resolve stasis and generate blood, thus harmonizing Yin and Yang, which can reach thoroughfare and conception vessels to nourish the uterus and recover the normal physiological function of the uterus. In that case, normal pregnancy is possible. Professor MA Kun attaches importance to the therapeutic principle of supplementing Qi and nourishing blood. In addition, she advocates conforming to changes in the menstrual cycle to promote the development of the uterus and the implantation of fertilized eggs. She also integrates traditional Chinese medicine and western medicine to treat both symptoms and root causes. Professor MA Kun's experience has demonstrated definite clinical effect on this disease and can be taken as a reference.
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Medicamentos Herbarios Chinos , Infertilidad Femenina , Riñón , Femenino , Humanos , Medicamentos Herbarios Chinos/uso terapéutico , Infertilidad Femenina/etiología , Infertilidad Femenina/tratamiento farmacológico , Útero/anomalías , Adulto , Medicina Tradicional China , Embarazo , Enfermedades Renales/etiología , Enfermedades Renales/tratamiento farmacológico , Anomalías UrogenitalesRESUMEN
Uterine fibroids are a prevalent factor that impacts fertility in women of reproductive age. This study discusses the theoretical foundation and formula principles of Professor MA Kun's clinical treatment for infertility caused by uterine fibroids. The kidney stores essence and is responsible for reproduction, while blood serves as a vital material basis for women's physiological functions. Kidney deficiency is the fundamental pathogenesis of infertility, and imbalances in kidney Qi and essence or deficiencies in kidney Yin and Yang can result in blood stasis. Blood stasis plays a significant role throughout this condition by impeding the flow of blood, which is crucial for nourishing Qi. Therefore, both kidney deficiency and blood stasis are key factors contributing to infertility caused by uterine fibroids. Professor MA Kun treats infertility caused by uterine fibroids using an approach that involves tonifying the kidneys and activating blood circulation based on changes in Qi and blood during the menstrual cycle as well as follicular growth processes. By identifying stage-specific evidence, appropriate treatments can be applied accordingly. During menstruation when the uterus opens and menstrual blood flows out, promoting follicular development through nourishing kidney Yin and activating blood circulation becomes essential. In later stages of menstruation, additional measures are taken to remove blood stasis, alleviate symptoms, disperse knots, attack pathogens while simultaneously replenishing vital energy. During intermenstrual periods when Yin holds greater importance than Yang, tonifying the kidneys and activating blood circulation helps facilitate smooth discharge of eggs by promoting transformation between Yin and Yang energies. Premenstrual period to warm kidney Yang to promote pregnant egg implantation, and at the same time to dredge the liver and regulate Qi, Qi elimination stagnation, complementary in the line, with the symptoms of additional subtractions. Clinical effect is remarkable, for the reference of colleagues.
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Medicamentos Herbarios Chinos , Infertilidad Femenina , Riñón , Leiomioma , Humanos , Femenino , Riñón/fisiopatología , Infertilidad Femenina/etiología , Infertilidad Femenina/terapia , Infertilidad Femenina/fisiopatología , Medicamentos Herbarios Chinos/uso terapéuticoRESUMEN
Quantum key distribution (QKD) uses individual light quanta in quantum superposition states to guarantee unconditional communication security between distant parties. However, the distance over which QKD is achievable has been limited to a few hundred kilometres, owing to the channel loss that occurs when using optical fibres or terrestrial free space that exponentially reduces the photon transmission rate. Satellite-based QKD has the potential to help to establish a global-scale quantum network, owing to the negligible photon loss and decoherence experienced in empty space. Here we report the development and launch of a low-Earth-orbit satellite for implementing decoy-state QKD-a form of QKD that uses weak coherent pulses at high channel loss and is secure because photon-number-splitting eavesdropping can be detected. We achieve a kilohertz key rate from the satellite to the ground over a distance of up to 1,200 kilometres. This key rate is around 20 orders of magnitudes greater than that expected using an optical fibre of the same length. The establishment of a reliable and efficient space-to-ground link for quantum-state transmission paves the way to global-scale quantum networks.
RESUMEN
Non-healing diabetic wounds (DW) are a serious clinical problem that remained poorly understood. We recently found that topical application of growth differentiation factor 11 (GDF11) accelerated skin wound healing in both Type 1 DM (T1DM) and genetically engineered Type 2 diabetic db/db (T2DM) mice. In the present study, we elucidated the cellular and molecular mechanisms underlying the action of GDF11 on healing of small skin wound. Single round-shape full-thickness wound of 5-mm diameter with muscle and bone exposed was made on mouse dorsum using a sterile punch biopsy 7 days following the onset of DM. Recombinant human GDF11 (rGDF11, 50 ng/mL, 10 µL) was topically applied onto the wound area twice a day until epidermal closure (maximum 14 days). Digital images of wound were obtained once a day from D0 to D14 post-wounding. We showed that topical application of GDF11 accelerated the healing of full-thickness skin wounds in both type 1 and type 2 diabetic mice, even after GDF8 (a muscle growth factor) had been silenced. At the cellular level, GDF11 significantly facilitated neovascularization to enhance regeneration of skin tissues by stimulating mobilization, migration and homing of endothelial progenitor cells (EPCs) to the wounded area. At the molecular level, GDF11 greatly increased HIF-1É expression to enhance the activities of VEGF and SDF-1É, thereby neovascularization. We found that endogenous GDF11 level was robustly decreased in skin tissue of diabetic wounds. The specific antibody against GDF11 or silence of GDF11 by siRNA in healthy mice mimicked the non-healing property of diabetic wound. Thus, we demonstrate that GDF11 promotes diabetic wound healing via stimulating endothelial progenitor cells mobilization and neovascularization mediated by HIF-1É-VEGF/SDF-1É pathway. Our results support the potential of GDF11 as a therapeutic agent for non-healing DW.
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Diabetes Mellitus Experimental , Células Progenitoras Endoteliales , Factores de Diferenciación de Crecimiento , Cicatrización de Heridas , Animales , Humanos , Ratones , Proteínas Morfogenéticas Óseas/metabolismo , Quimiocina CXCL12/efectos de los fármacos , Quimiocina CXCL12/metabolismo , Diabetes Mellitus Experimental/complicaciones , Diabetes Mellitus Experimental/tratamiento farmacológico , Diabetes Mellitus Experimental/metabolismo , Diabetes Mellitus Tipo 2/tratamiento farmacológico , Diabetes Mellitus Tipo 2/metabolismo , Células Progenitoras Endoteliales/metabolismo , Células Progenitoras Endoteliales/patología , Factores de Diferenciación de Crecimiento/uso terapéutico , Factores de Diferenciación de Crecimiento/metabolismo , Neovascularización Fisiológica , Factor A de Crecimiento Endotelial Vascular/efectos de los fármacos , Factor A de Crecimiento Endotelial Vascular/metabolismo , Cicatrización de Heridas/efectos de los fármacos , Proteínas Recombinantes/metabolismo , Proteínas Recombinantes/uso terapéutico , Subunidad alfa del Factor 1 Inducible por Hipoxia/efectos de los fármacos , Subunidad alfa del Factor 1 Inducible por Hipoxia/metabolismoRESUMEN
INTRODUCTION: This study aims to analyze the permeability of intra- and peri-meningiomas regions and compare the microvascular permeability between peritumoral brain edema (PTBE) and non-PTBE using DCE-MRI. METHODS: This was a retrospective of patients with meningioma who underwent surgery. The patients were grouped as PTBE and non-PTBE. The DCE-MRI quantitative parameters, including volume transfer constant (Ktrans), rate constant (Kep), extracellular volume (Ve), and mean plasma volume (Vp), obtained using the extended Tofts-Kety 2-compartment model. Logistic regression analysis was conducted to explore the risk factor of PTBE. RESULTS: Sixty-three patients, diagnosed as fibrous meningioma, were included in this study. They were 17 males and 46 females, aged from 32 to 88 years old. Kep and Vp were significantly lower in patients with PTBE compared with those without (Kep: 0.1852 ± 0.0369 vs. 0.5087 ± 0.1590, p = 0.010; Vp: 0.0090 ± 0.0020 vs. 0.0521 ± 0.0262, p = 0.007), while there were no differences regarding Ktrans and Ve (both p > 0.05). The multivariable analysis showed that tumor size ≥10 cm3 (OR = 4.457, 95% CI: 1.322-15.031, p = 0.016) and Vp (OR = 0.572, 95%CI: 0.333-0.981, p = 0.044) were independently associated with PTBE in patients with meningiomas. CONCLUSION: DCE-magnetic resonance imaging·Meningioma·Blood vessel MRI can be used to quantify the microvascular permeability of PTBE in patients with meningioma.
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Edema Encefálico , Neoplasias Meníngeas , Meningioma , Adulto , Anciano , Anciano de 80 o más Años , Edema Encefálico/diagnóstico por imagen , Edema Encefálico/etiología , Permeabilidad Capilar , Medios de Contraste , Femenino , Humanos , Imagen por Resonancia Magnética , Masculino , Neoplasias Meníngeas/diagnóstico por imagen , Meningioma/diagnóstico por imagen , Persona de Mediana Edad , Estudios RetrospectivosRESUMEN
BACKGROUND: Chronic brain hypoperfusion (CBH) is closely related to Alzheimer's disease (AD) and vascular dementia (VaD). Meanwhile, synaptic pathology plays a prominent role in the initial stage of AD and VaD. However, whether and how CBH impairs presynaptic plasticity is currently unclear. METHODS: In the present study, we performed a battery of techniques, including primary neuronal culture, patch clamp, stereotaxic injection of the lentiviral vectors, morris water maze (MWM), dual luciferase reporter assay, FM1-43 fluorescence dye evaluation, qRT-PCR and western blot, to investigate the regulatory effect of miR-153 on hippocampal synaptic vesicle release both in vivo and in vitro. The CBH rat model was generated by bilateral common carotid artery ligation (2VO). RESULTS: Compared to sham rats, 2VO rats presented decreased field excitatory postsynaptic potential (fEPSP) amplitude and increased paired-pulse ratios (PPRs) in the CA3-CA1 pathway, as well as significantly decreased expression of multiple vesicle fusion-related proteins, including SNAP-25, VAMP-2, syntaxin-1A and synaptotagmin-1, in the hippocampi. The levels of microRNA-153 (miR-153) were upregulated in the hippocampi of rats following 2VO surgery, and in the plasma of dementia patients. The expression of the vesicle fusion-related proteins affected by 2VO was inhibited by miR-153, elevated by miR-153 inhibition, and unchanged by binding-site mutation or miR masks. FM1-43 fluorescence images showed that miR-153 blunted vesicle exocytosis, but this effect was prevented by either 2'-O-methyl antisense oligoribonucleotides to miR-153 (AMO-153) and miR-masking of the miR-153 binding site in the 3' untranslated region (3'UTR) of the Snap25, Vamp2, Stx1a and Syt1 genes. Overexpression of miR-153 by lentiviral vector-mediated miR-153 mimics (lenti-pre-miR-153) decreased the fEPSP amplitude and elevated the PPR in the rat hippocampus, whereas overexpression of the antisense molecule (lenti-AMO-153) reversed these changes triggered by 2VO. Furthermore, lenti-AMO-153 attenuated the cognitive decline of 2VO rats. CONCLUSIONS: Overexpression of miR-153 controls CBH-induced presynaptic vesicle release impairment by posttranscriptionally regulating the expression of four vesicle release-related proteins by targeting the 3'UTRs of the Stx1a, Snap25, Vamp2 and Syt1 genes. These findings identify a novel mechanism of presynaptic plasticity impairment during CBH, which may be a new drug target for prevention or treatment of AD and VaD. Video Abstract.
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Demencia Vascular/metabolismo , Hipoxia-Isquemia Encefálica/metabolismo , MicroARNs/fisiología , Vesículas Sinápticas/metabolismo , Anciano , Animales , Humanos , Masculino , Ratas , Ratas Sprague-Dawley , Proteína 25 Asociada a Sinaptosomas/metabolismo , Sinaptotagmina I/metabolismo , Sintaxina 1/metabolismo , Proteína 2 de Membrana Asociada a Vesículas/metabolismoRESUMEN
Chronic brain hypoperfusion (CBH) is a common clinical feature of Alzheimer's disease and vascular dementia, but the underlying molecular mechanism is unclear. Our previous study reported that the down-regulation of microRNA-195 (miR-195) promotes amyloidogenesis via regulation of amyloid precursor protein and ß-site amyloid precursor protein cleaving enzyme 1 (BACE1) expression at the post-transcriptional level in CBH rats with bilateral common carotid artery occlusion (2VO). CBH owing to unilateral common carotid artery occlusion (UCCAO) increases tau phosphorylation levels at multiple phosphorylation sites in the brain, but the molecular mechanism is poorly understood. The purpose of this study was to investigate whether miR-195 could both deregulate amyloid metabolism and indirectly deregulate tau phosphorylation in CBH. We observed that 2VO leads to tau hyperphosphorylation at Ser202/Thr205, Ser262, Thr231, and Ser422 and to the conversion from cyclin-dependent kinase 5 (Cdk5)/p35 to Cdk5/p25 in rat hippocampi. Endogenous miR-195 was knocked down using over-expression of its antisense molecule (pre-AMO-miR-195) via a lentivirus (lenti-pre-AMO-miR-195); this knockdown increased the tau phosphorylation at Ser202/Thr205, Ser262, Thr231, Ser422, and the Cdk5/p25 activation, but over-expression of miR-195 using lenti-pre-miR-195 decreased the tau phosphorylation and Cdk5/p25 activation. Further in vitro studies demonstrated that miR-195 over-expression prevented tau hyperphosphorylation and Cdk5/p35 activity, which were increased by miR-195 inhibition. A dual luciferase reporter assay showed that miR-195 bound to the Cdk5r1 gene, which encodes p35 protein, in the 3'UTR and inhibited p35 expression. We concluded that tau hyperphosphorylation involves the down-regulation of miR-195, which is mediated by Cdk5/p25 activation in 2VO rats. Our findings demonstrated that down-regulation of miR-195 led to increased vulnerability via the regulation of multiple targets. Schematic diagram of miR-195 mediated Aß aggregation and tau hyperphosphorylation in chronic brain hypoperfusion (CBH). First, CBH results in the elevation of nuclear factor-κB (NF-κB), which binds with the promoter sequences of miR-195 and negatively regulates the expression of miR-195. Second, down-regulated miR-195 induces up-regulation of APP and BACE1 and leads to an increase in Aß levels. Third, some of the elevated Aß then enter the intracellular space and activate calpain, which promotes the conversion of Cdk5/p35 to Cdk5/p25 and catalyzes the degradation of IκB; IκB is an inhibitor of NF-κB, which activates NF-κB. Cdk5/p25 directly phosphorylates Tau. Fourth, down-regulated miR-195 induces an up-regulation of p35, which provides the active substrates of p25. Our findings demonstrated that the down-regulation of miR-195 plays a key role in the increased vulnerability to dementia via the regulation of multiple targets following CBH.
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Enfermedad de Alzheimer/metabolismo , Isquemia Encefálica/metabolismo , Quinasa 5 Dependiente de la Ciclina/metabolismo , MicroARNs/metabolismo , Proteínas tau/metabolismo , Animales , Western Blotting , Encéfalo/irrigación sanguínea , Encéfalo/metabolismo , Isquemia Encefálica/complicaciones , Enfermedad Crónica , Modelos Animales de Enfermedad , Regulación hacia Abajo , Masculino , Fosforilación , Ratas , Ratas Sprague-Dawley , Reacción en Cadena en Tiempo Real de la Polimerasa , TransfecciónRESUMEN
Previous studies have demonstrated that chronic brain hypoperfusion (CBH) causes Aß aggregation by upregulating expression of amyloid precursor protein (APP) and ß-site APP cleaving enzyme 1 (BACE1) protein, which is accompanied by cognitive impairment, but the mechanisms are not fully understood. In this study, we evaluated the effect of microRNA on memory impairment in rats induced by CBH. We show here that CBH generated by bilateral common carotid artery occlusion (2VO) significantly decreased the learning and memory ability in rats, as assessed by Morris water maze, and upregulated expression of APP and BACE1 proteins in the hippocampus and cortex of rats, as evaluated by Western blot and immunofluorescence. In reciprocal, qRT-PCR analysis showed that microRNA-195 (miR-195) was downregulated in both the hippocampus and cortex of rats following CBH, and in the plasma of dementia patients. APP and BACE1 proteins were downregulated by miR-195 overexpression, upregulated by miR-195 inhibition, and unchanged by binding-site mutation or miR-masks, indicating that APP and BACE1 are two potential targets for miR-195. Knockdown of endogenous miR-195 by lentiviral vector-mediated overexpression of its antisense molecule (lenti-pre-AMO-miR-195) elicited dementia in rats, whereas overexpression of miR-195 using lenti-pre-miR-195 reduced dementia vulnerability triggered by 2VO. Additionally, chromatin immunoprecipitation analysis showed that NFκB was bound to the promoter region of miR-195 and inhibited its expression. We conclude that miR-195 may play a key role in determining dementia susceptibility in 2VO rats by regulating APP and BACE1 expression at the post-transcriptional level, and exogenous complement of miR-195 may be a potentially valuable anti-dementia approach.
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Péptidos beta-Amiloides/metabolismo , Enfermedades de las Arterias Carótidas/complicaciones , Demencia/etiología , Regulación de la Expresión Génica/fisiología , MicroARNs/uso terapéutico , Fármacos Neuroprotectores/uso terapéutico , Anciano , Anciano de 80 o más Años , Secretasas de la Proteína Precursora del Amiloide/genética , Secretasas de la Proteína Precursora del Amiloide/metabolismo , Péptidos beta-Amiloides/sangre , Precursor de Proteína beta-Amiloide/genética , Precursor de Proteína beta-Amiloide/metabolismo , Análisis de Varianza , Animales , Animales Recién Nacidos , Ácido Aspártico Endopeptidasas/genética , Ácido Aspártico Endopeptidasas/metabolismo , Células Cultivadas , Corteza Cerebral/citología , Corteza Cerebral/metabolismo , Demencia/genética , Demencia/patología , Demencia/terapia , Modelos Animales de Enfermedad , Ensayo de Inmunoadsorción Enzimática , Femenino , Regulación de la Expresión Génica/efectos de los fármacos , Hipocampo/citología , Hipocampo/metabolismo , Humanos , Inmunoprecipitación , Lipopolisacáridos/farmacología , Masculino , Aprendizaje por Laberinto/efectos de los fármacos , Aprendizaje por Laberinto/fisiología , MicroARNs/biosíntesis , MicroARNs/sangre , Neuronas/efectos de los fármacos , Fármacos Neuroprotectores/metabolismo , Oligonucleótidos Antisentido/farmacología , Fragmentos de Péptidos/metabolismo , Proteínas Serina-Treonina Quinasas/genética , Proteínas Serina-Treonina Quinasas/metabolismo , ARN Mensajero/metabolismo , Ratas , Ratas Sprague-Dawley , Transfección , Quinasa de Factor Nuclear kappa BRESUMEN
It is well known that menopause could worsen age-related ventricular concentric remodeling following estrogen (E2) deficiency. However the underlying mechanisms of such phenomena are not fully understood. Mitochondria, as the 'cellular power station' of hearts, play an important role in maintaining normal cardiac function and structure. Therefore, the present study aims to investigate whether mitochondrial compromise is responsible for E2 deficiency associated concentric remodeling and, if so, what is its underlying molecular mechanism. We found evident concentric remodeling pattern in both postmenopausal and ovariectomized (OVX) mice, which could be attenuated by E2 replacement. Further study showed mitochondrial structural damages and respiratory function impairment in myocardium of both postmenopausal and OVX mice and E2 supplement reversed mitochondrial dysfunction in OVX mice, suggesting that E2 deficiency could induce mitochondrial compromise in the heart. Then, peroxisome proliferator-activated receptor-γ co-activator 1-α (PGC-1α), a key mitochondrial function and biology regulator, was found significantly reduced in both postmenopausal and OVX mice. The reduction of PGC-1α protein level in OVX mice could be rescued by E2 delivery, indicating that E2 could positively regulate PGC-1α expression. Next, we found that microRNA-23a (miR-23a) could be negatively regulated by E2 in both myocardium and cultured cardiomyocytes. Moreover, miR-23a could directly downregulate PGC-1α expression in cardiomyocytes via binding to its 3'UTR which implied that miR-23a could be critical for the downregulation of PGC-1α under E2 deficiency. Overexpression of miR-23a was also found to damage mitochondria in cultured cardiomyocytes, ascribed to PGC-1α downregulation. Taken together, E2 deficiency may cause mitochondrial compromise through miR-23a-mediated PGC-1α downregulation, which may subsequently lead to the menopause-associated concentric remodeling.
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Estrógenos/deficiencia , MicroARNs/metabolismo , Mitocondrias Cardíacas/metabolismo , Factores de Transcripción/metabolismo , Remodelación Ventricular , Animales , Animales Recién Nacidos , Secuencia de Bases , Respiración de la Célula , Regulación hacia Abajo , Estrógenos/metabolismo , Femenino , Ratones Endogámicos C57BL , MicroARNs/genética , Mitocondrias Cardíacas/ultraestructura , Datos de Secuencia Molecular , Miocardio/metabolismo , Miocardio/patología , Miocitos Cardíacos/metabolismo , Coactivador 1-alfa del Receptor Activado por Proliferadores de Peroxisomas gamma , PosmenopausiaRESUMEN
BACKGROUND: Aging is associated with the gradual cognitive decline and shows the typical senile plaque formation in the brain, which results from the aggregation of beta amyloid (Aß) peptide following the abnormal proteolytic processing of amyloid precursor protein (APP) by ß-secretase (BACE1) and γ-secretase. Accumulating evidence indicates that several microRNAs (miRNAs) are involved in the Alzheimer's disease (AD) by regulating the expression of APP and BACE1 proteins. However, the cognitive ability and the expression profile of the APP- and BACE1-associated miRNAs in the middle-aged population are largely unknown. METHODS: The learning and memory ability in rats were determined by Morris Water Maze test. The protein levels of APP and BACE1 were detected by western blotting. The quantitative polymerase chain reaction was used to identify the miRNAs levels in forebrain cortex and the hippocampus. RESULTS: Middle-aged rats have declined learning ability without changes in the memory ability, and increased APP and BACE1 protein expression in the forebrain cortex. Computational analysis using Targetscan and Pictar databases reveals that totally 4 predicted miRNAs have conserved binding site with APP, namely miR-106b, -17-5p, -153, -101. All of them showed decreased expression in both the forebrain cortex and hippocampus. Among the 10 predicted miRNAs targeting BACE1, different expression profiles were identified in the forebrain cortex (decreased: miR-9, -19a, -135a, -15b, -16, -195, -29c, -214; increased: miR-124; no change: miR-141) and the hippocampus (decreased: miR-9, -15b, -16, -195, -29c, -124; increased: miR-19a, -135a, -214, -141) in the middle-aged rats compared with the young rats. CONCLUSION: Our results provided the first evidence that middle-aged rats have begun displaying cognitive disability with abnormal expression of APP- and BACE1-related miRNAs in the hippocampus and forebrain cortex.
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Envejecimiento/genética , Amiloide/metabolismo , Regulación del Desarrollo de la Expresión Génica , Hipocampo/metabolismo , MicroARNs/genética , Secretasas de la Proteína Precursora del Amiloide/genética , Secretasas de la Proteína Precursora del Amiloide/metabolismo , Precursor de Proteína beta-Amiloide/genética , Precursor de Proteína beta-Amiloide/metabolismo , Animales , Ácido Aspártico Endopeptidasas/genética , Ácido Aspártico Endopeptidasas/metabolismo , Cognición , Perfilación de la Expresión Génica , Masculino , MicroARNs/metabolismo , ARN Mensajero/genética , ARN Mensajero/metabolismo , Ratas WistarRESUMEN
OBJECTIVE: To provide scientific evidence for medical insurance and health policies allocating the limited health resources in China. METHODS: Based on the data of the national household survey by the State Council Pilot Urban Resident Basic Medical Insurance Evaluation from Nov.2007 to Nov.2011, a two-step model and the human capital method were used to estimate the economic burden of five major chronic diseases among urban residents in China. According to the economic burden, the relative economic risk (relative risk, RR) and adjusted RR were calculated. The five chronic diseases were hypertension, cardiovascular, diabetes, arthritis or rheumatism and chronic lung diseases. RESULTS: More than 50% of the residents with these five chronic diseases were the over 65-year-old and retired. 90% of the residents with these five chronic diseases had medical insurance except the residents with chronic lung diseases. Average co-pay from the outpatient department and the pharmacy was more than 60%, and about 50% from the inpatient department. Annual total cost per capita was the highest 8 954.29 Yuan among the residents with cardiovascular disease and the second highest 8 914.36 Yuan among the residents with diabetes. The adjusted RR of the residents with cardiovascular and diabetes were greater than 1, respectively 1.36 and 1.15. CONCLUSION: The retired take up the largest percentage of population with chronic diseases, and the influence of the major five chronic diseases is more serious in north-west China. The main expenditure is from the outpatient department and the pharmacy, in which the availability of drugs reimbursed needs to be improved. The patients with cardiovascular and diabetes experience both higher economic burden and economic risk.
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Enfermedad Crónica/economía , Costo de Enfermedad , Artritis/economía , Enfermedades Cardiovasculares/economía , China , Ciudades , Costos y Análisis de Costo , Diabetes Mellitus/economía , Gastos en Salud , Humanos , Hipertensión/economía , Enfermedades Pulmonares/economíaRESUMEN
Surveillance of drug safety is an important aspect in the routine medical care. Adverse events caused by real-world drug utilization has become one of the leading causes of death and an urgent issue in the field of toxicology. Cardiovascular disease is now the leading cause of fatal diseases in most countries, especially in the elderly population who often suffer from multiple diseases and need long-term multidrug therapy. Among which, statins have been widely used to lower bad cholesterol and regress coronary plaque mainly in patients with hyperlipidemia and atherosclerotic cardiovascular diseases (ASCVD). Although the real-world benefits of statins are significant, different degrees and types of adverse drug reactions (ADR) such as liver dysfunction and muscle injury, have a great impact on the original treatment regimens as well as the quality of life. This review describes the epidemiology, mechanisms, early identification and post-intervention of statin-associated liver dysfunction and muscle injury based on the updated clinical evidence. It provides systematic and comprehensive guidance and necessary supplement for the clinical safety of statin use in cardiovascular diseases.
RESUMEN
High-quality crystals are key to obtaining accurate three-dimensional structures of proteins using X-ray diffraction techniques. However, obtaining such protein crystals is often a challenge. Several containerless crystallization techniques have been reported to have the ability to improve crystal quality, but it is unknown which is the most favourable way to grow high-quality protein crystals. In this paper, a quality comparison of protein crystals which were grown under three containerless conditions provided by diamagnetic levitation, silicone oil and agarose gel was conducted. A control experiment on a vessel wall was also simultaneously carried out. Seven different proteins were crystallized under the four conditions, and the crystal quality was assessed in terms of the resolution limit, the mosaicity and the Rmerge. It was found that the crystals grown under the three containerless conditions demonstrated better morphology than those of the control. X-ray diffraction data indicated that the quality of the crystals grown under the three containerless conditions was better than that of the control. Of the three containerless crystallization techniques, the diamagnetic levitation technique exhibited the best performance in enhancing crystal quality. This paper is to our knowledge the first report of improvement of crystal quality using a diamagnetic levitation technique. Crystals obtained from agarose gel demonstrated the second best improvement in crystal quality. The study indicated that the diamagnetic levitation technique is indeed a favourable method for growing high-quality protein crystals, and its utilization is thus potentially useful in practical efforts to obtain well diffracting protein crystals.
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Cristalografía por Rayos X , Gravitación , Espectroscopía de Resonancia Magnética , Espectroscopía de Fotoelectrones , Proteínas/química , Sefarosa/normas , Aceites de Silicona/normas , Animales , Pollos , Cristalización/métodos , Cristalización/normas , Cristalografía por Rayos X/métodos , Cristalografía por Rayos X/normas , Proteínas de Escherichia coli/química , Proteínas/normas , Control de Calidad , Trichosanthes , Difracción de Rayos X/métodos , Difracción de Rayos X/normasRESUMEN
BACKGROUND/AIMS: Abnormal baroreceptor reflex sensitivity (BRS) and elevated plasma neuropeptide Y (NPY) are prevalent in diabetic patients. The present study was conducted to determine whether NPY Y1 receptor (Y1R) and NPY Y2 receptor (Y2R) contribute to the regulatin of BRS in diabetic rats. METHODS: Diabetes mellitus (DM) rats with hyperlipidemia were developed by an emulsion diet enriched with fat, sucrose and fructose followed by streptozocin (STZ). Y1R and Y2R specific antagonists (BIBP 3226 and BIIE 0246) were administered by a mini-osmotic pump. Systolic blood pressure (SBP), heart rate (HR), BRS and heart functions, as well as the plasma NPY and lipid level were measured after treatment for 4 weeks. RESULTS: Both BIBP 3226 and BIIE 0246 treatment reversed the elevated total cholesterol (TC) and low density lipoprotein (LDL-C) level, and reduced high density lipoprotein (HDL-C) level in DM rats. BIIE 0246 may attenuate the increased triglyceride (TG) level in DM rats. In addition, neither BIBP 3226 nor BIIE 0246 treatment produced significant effects on BRS, SBP or HR (P>0.05) in DM rats, even after PE and SNP challenge. However, BIBP 3226 and BIIE 0246 further impaired LVSP, LVEDP, +dp/dtmax and -dp/dtmax. CONCLUSION: This study provided us with the evidence that the inhibition of peripheral Y1R and Y2R did not affect impaired BRS but amplified the deterioration of the compromised cardiac function in STZ-induced DM rats with hyperlipidemia.
Asunto(s)
Barorreflejo/fisiología , Diabetes Mellitus Experimental/metabolismo , Receptores de Neuropéptido Y/antagonistas & inhibidores , Animales , Arginina/análogos & derivados , Arginina/farmacología , Barorreflejo/efectos de los fármacos , Benzazepinas/farmacología , Presión Sanguínea/efectos de los fármacos , Colesterol/sangre , LDL-Colesterol/sangre , Diabetes Mellitus Experimental/complicaciones , Diabetes Mellitus Experimental/patología , Frecuencia Cardíaca/efectos de los fármacos , Hiperlipidemias/complicaciones , Hiperlipidemias/metabolismo , Hiperlipidemias/patología , Masculino , Neuropéptido Y/sangre , Nitroprusiato/farmacología , Fenilefrina/farmacología , Ratas , Ratas Wistar , Receptores de Neuropéptido Y/metabolismo , Triglicéridos/sangreRESUMEN
OBJECTIVE: To investigate the characteristics of pulmonary function and respiratory muscle performance in patients with multiple system atrophy (MSA) and Parkinson's disease (PD). METHODS: Pulmonary function and respiratory muscle strength were evaluated in 16 MSA patients and 20 PD patients. Another 17 age and sex-matched healthy volunteers were recruited as controls. RESULTS: Carbon monoxide diffusion capacity (DLCO) was significantly decreased in MSA group compared with PD group [(62.86 ± 15.66)% vs (76.67 ± 18.98)%, respectively, P < 0.05]. Maximal inspiration pressure (MIP) and maximal expiration pressure (MEP) were (39.08 ± 22.99)% and (49.59 ± 22.97)% in MSA group, (39.83 ± 24.83)% and (49.26 ± 22.86)% in PD group. Both MIP and MEP in MSA and PD groups were significantly reduced compared with controls [(57.44 ± 19.90)%, P < 0.05; (77.10 ± 21.46)%, P < 0.001]. CONCLUSIONS: Our findings suggest that respiratory dysfunction is involved in MSA and PD. The reduction of respiratory muscle strength is remarkable. The insufficiency of pulmonary diffusion function is more severe in MSA than in PD. More attention should be paid to the compromised respiratory function in neurodegenerative disorders.
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Atrofia de Múltiples Sistemas/fisiopatología , Fuerza Muscular , Enfermedad de Parkinson/fisiopatología , Músculos Respiratorios/fisiopatología , Anciano , Estudios de Casos y Controles , Femenino , Humanos , Pulmón/fisiopatología , Masculino , Persona de Mediana Edad , Pruebas de Función RespiratoriaRESUMEN
OBJECTIVE: To observe clinical and pathological features of nephrogenic adenoma (NA), and to find some useful immunohistochemical markers for its diagnosis. METHODS: The clinical features of 32 NA patients were obtained. Each case underwent microscopic observation and immumohistochemical staining. The primary antibodies were α-methylacyl-CoA racemase (AMACR, P504S), cytokeratin AE1/AE3, cytokeratin 7 (CK7), cytokeratin 20 (CK20), paired-box 2 (PAX2), paired-box 8 (PAX8), vimentin, membrane metallo-endopeptidase (MME, CD10), prostate specific antigen (PSA), high molecular weight cytokeratin (34ßE12), P63 and carcinoembryonic antigen (CEA). RESULTS: NA mainly involved old men, and the bladder was the commonest location. The macroscopic features were prevalently small polypoid or papillary lesions, ranging from 1 mm to 10 mm (mean=4). The typical histological features included tubular, tubulocystic, polypoid and/or papillary. Immunohistochemistry for NA was positive for AMACR, AE1/AE3, PAX2, PAX8, CK7, vimentin and CD10. The negative immunostain for NA included P63, PSA and CEA. CONCLUSION: NA is a rare and easily misdiagnosed lesion. Careful histological examination is essential to accurately identify this lesion. A panel composed of AMACR (P504S), PAX8/PAX2, CK7, P63, PSA and CEA appears to be sensitive and specific in differentiating NA from its mimics of urothelial and prostatic origins.
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Adenoma/patología , Biomarcadores de Tumor , Adenoma/diagnóstico , Adenoma/inmunología , Antígeno Carcinoembrionario , Humanos , Inmunohistoquímica , Riñón/patología , Antígeno Prostático Específico , Racemasas y EpimerasasRESUMEN
Tight junctions (TJs) are located in the apical region of the junctions between epithelial cells and are widely found in organs such as the brain, retina, intestinal epithelium, and endothelial system. As a mechanical barrier of the intestinal mucosa, TJs can not only maintain the integrity of intestinal epithelial cells but also maintain intestinal mucosal permeability by regulating the entry of ions and molecules into paracellular channels. Therefore, the formation disorder or integrity destruction of TJs can induce damage to the intestinal epithelial barrier, ultimately leading to the occurrence of various gastrointestinal diseases, such as inflammatory bowel disease (IBD), gastroesophageal reflux disease (GERD), and irritable bowel syndrome (IBS). However, a large number of studies have shown that TJs protein transport disorder from the endoplasmic reticulum to the apical membrane can lead to TJs formation disorder, in addition to disruption of TJs integrity caused by external pathological factors and reduction of TJs protein synthesis. In this review, we focus on the structural composition of TJs, the formation of clathrin-coated vesicles containing transmembrane TJs from the Golgi apparatus, and the transport process from the Golgi apparatus to the plasma membrane via microtubules and finally fusion with the plasma membrane. At present, the mechanism of the intracellular transport of TJ proteins remains unclear. More studies are needed in the future to focus on the sorting of TJs protein vesicles, regulation of transport processes, and recycling of TJ proteins, etc.
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Intestinos , Proteínas de Uniones Estrechas , Proteínas de Uniones Estrechas/metabolismo , Mucosa Intestinal/metabolismo , Células Epiteliales/metabolismo , Uniones Estrechas/metabolismoRESUMEN
Enteric glial cells (EGC) play an essential role in maintaining the integrity of intestinal epithelial barrier (IEB). However, the mechanism of EGCs in the regulation of IEB functions under lipopolysaccharide (LPS) stimulation is unknown. To investigate the barrier-related role of EGCs in response to the LPS challenge, the coculture model of EGCs and intestinal epithelial cells (IEC) IEC-6 was established in vitro. Transepithelial resistance (TER) measurements showed that, LPS treatment significantly increased barrier permeability of IEC monolayer from the basolateral side (35.4±6.3 Ω/cm(2), p<0.05) but not the apical side (69.7±6.3 Ω/cm(2)) when compared with the control group (81.8±10.9 Ω/cm(2)). The assessment of intestinal epithelial integrity by TER reading and by measuring expression of tight junction protein revealed that, incubation with EGCs or EGC conditioned media significantly increased the TER of IEC monolayers under normal condition as well as the LPS stimulation, accompanied with upregulating zonula occludens-1 and occludin expression at mRNA and protein levels. Real-time quantitative polymerase chain reaction and nitric production assay demonstrated that LPS exposure elicited a maximally 13-fold increase of inducible nitric oxide synthase (iNOS) mRNA expression and 10-fold increase of nitric oxide production of EGCs. After being pretreated with the selective iNOS inhibitor 1400 W, EGCs significantly increased the TER of IEC monolayers against the disruption effect of LPS (p<0.05). These findings suggest that EGCs play an important role in maintaining the IEB function in response to the LPS stimulation. The protective effect of EGCs on IEB functions could be enhanced by inhibiting the increase of iNOS activity induced by LPS.
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Mucosa Intestinal/citología , Mucosa Intestinal/metabolismo , Neuroglía/citología , Óxido Nítrico Sintasa de Tipo II/metabolismo , Animales , Western Blotting , Línea Celular , Técnicas de Cocultivo , Sistema Nervioso Entérico/citología , Técnica del Anticuerpo Fluorescente , Lipopolisacáridos/toxicidad , Neuroglía/metabolismo , Permeabilidad , Ratas , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Uniones Estrechas/metabolismoRESUMEN
OBJECTIVE: Severe cases of coronavirus disease 2019 (COVID-19) are expected to have a worse prognosis than mild cases. Shenhuang Granule (SHG) has been shown to be a safe and effective treatment for severe COVID-19 in a previous randomized clinical trial, but the active chemical constituents and underlying mechanisms of action remain unknown. The goal of this study is to explore the chemical basis and mechanisms of SHG in the treatment of severe COVID-19, using network pharmacology. METHODS: Ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry was employed to screen chemical constituents of SHG. Putative therapeutic targets were predicted by searching traditional Chinese medicine system pharmacology database and analysis platform, SwissTargetPrediction, and Gene Expression Omnibus (GEO) databases. The target protein-protein interaction network and enrichment analysis were performed to investigate the hub genes and presumptive mechanisms. Molecular docking and molecular dynamics simulations were used to verify the stability and interaction between the key chemical constituents of SHG and COVID-19 protein targets. RESULTS: Forty-five chemical constituents of SHG were identified along with 131 corresponding therapeutic targets, including hub genes such as HSP90AA1, MMP9, CXCL8, PTGS2, IFNG, DNMT1, TYMS, MDM2, HDAC3 and ABCB1. Functional enrichment analysis indicated that SHG mainly acted on the neuroactive ligand-receptor interaction, calcium signaling pathway and cAMP signaling pathway. Molecular docking showed that the key constituents had a good affinity with the severe acute respiratory syndrome coronavirus 2 protein targets. Molecular dynamics simulations indicated that ginsenoside Rg4 formed a stable protein-ligand complex with helicase. CONCLUSION: Multiple components of SHG regulated multiple targets to inhibit virus invasion and cytokine storm through several signaling pathways; this provides a scientific basis for clinical applications and further experiments.