Exercise fatigue injury under sport resistance / Lesão de fadiga causada por exercício sob resistência esportiva / Lesión por fatiga causada por ejercicio bajo resistencia deportiva

ABSTRACT Introduction Athletes are prone to local muscle fatigue due to high-intensity training and to long-term accumulation of musculoskeletal injuries. Musculoskeletal complications represent a large proportion of occupational health problems and, for this reason, have received increased attention from the sports industry. In this sense, studies on muscle fatigue should be intensified. Objective Verify muscle fatigue and injury involving the strain characteristics of lower limb joints in the eccentric and centripetal contraction of the jump. Methods A total of 691 individuals aged 20 to 40 years were selected. Fatigue was caused by active muscle contraction. The characteristic curves of active muscle contraction in different isometric, isotonic, and isokinetic training were analyzed. The degree of fatigue caused by three different sports states was tested by experimentation. The corresponding active muscle contraction characteristics were also analyzed. The potential for homeostasis at different ages was compared. Results The delay in recovery to fatigue is directly proportional to the athlete's age. The return to post-exercise relief proportion from fatigue gradually decreases. Conclusion The experimental results showed that active muscle contraction could reduce exercise fatigue to some extent. This beneficial biochemical property of active muscle contraction is not found in people with advanced age. The findings have a guiding potential for the relief of sports fatigue. Evidence Level II; Therapeutic Studies - Investigating the result.

RESUMO Introdução Os esportistas estão propensos à fadiga muscular local devido ao treinamento de alta intensidade, e ao acúmulo de lesões musculoesqueléticas a longo prazo. As complicações musculoesqueléticas representam uma grande fatia dos problemas de saúde ocupacional e por isso têm recebido maior atenção da indústria esportiva. Nesse sentido, os estudos sobre a fadiga muscular devem ser aprofundados. Objetivo Verificar a fadiga e lesão muscular envolvendo as características de esforço das articulações dos membros inferiores na contração excêntrica e contração centrípeta do salto. Métodos Foram selecionados 691 indivíduos com idade entre 20 a 40 anos. A fadiga foi ocasionada por contração muscular ativa. Foram analisadas as curvas características da contração muscular ativa em diferentes estados nos treinos isométrico, isotônico e isocinético. O grau de fadiga causado por três estados esportivos diferentes foi testado através de experimentos. Também foram analisadas as características de contração muscular ativa correspondentes. O potencial de homeostase em diferentes idades foi comparado. Resultados O retardo na recuperação à fadiga é diretamente proporcional a idade do esportista. A proporção de retorno ao alívio pós-exercício sobre a fadiga diminui gradualmente. Conclusão Os resultados experimentais mostram que a contração muscular ativa pode reduzir até certo ponto a fadiga ao exercício. Os efeitos dessas propriedades bioquímicas benéficas da contração muscular ativa não são encontrados em pessoas com idade avançada. Os achados tem um potencial orientador para o alívio da fadiga esportiva. Nível de evidência II; Estudos Terapêuticos - Investigação de Resultados.

RESUMEN Introducción Los deportistas son propensos a la fatiga muscular local debido al entrenamiento de alta intensidad, y a la acumulación de lesiones musculoesqueléticas a largo plazo. Las complicaciones musculoesqueléticas representan una gran parte de los problemas de salud laboral y por ello han recibido una mayor atención por parte de la industria del deporte. En este sentido, hay que profundizar en los estudios sobre la fatiga muscular. Objetivo Verificar la fatiga muscular y las lesiones que implican las características de esfuerzo de las articulaciones de los miembros inferiores en la contracción excéntrica y centrípeta del salto. Métodos Se seleccionaron 691 individuos de entre 20 y 40 años. La fatiga fue causada por la contracción muscular activa. Fueron analizadas las curvas características de la contracción muscular activa en diferentes estados en el entrenamiento isométrico, isotónico e isocinético. El grado de fatiga provocado por tres estados deportivos diferentes se comprobó mediante experimentos. También se analizaron las correspondientes características de la contracción muscular activa. Se comparó el potencial de homeostasis a diferentes edades. Resultados El retraso en la recuperación a la fatiga es directamente proporcional a la edad del deportista. La proporción de retorno al alivio de la fatiga después del ejercicio disminuye gradualmente. Conclusión Los resultados experimentales muestran que la contracción muscular activa puede reducir la fatiga del ejercicio en cierta medida. Los efectos de estas propiedades bioquímicas beneficiosas de la contracción muscular activa no se encuentran en las personas mayores. Los hallazgos tienen un potencial orientador para el alivio de la fatiga deportiva. Nivel de evidencia II; Estudios terapéuticos - Investigación de resultados.

Integrated Network Pharmacology Analysis and Experimental Validation to Investigate the Molecular Mechanism of Triptolide in the Treatment of Membranous Nephropathy.

Background: Triptolide, a major active ingredient isolated from Tripterygium wilfordii Hook f., is effective in the treatment of membranous nephropathy (MN); however, its pharmacological mechanism of action has not yet been clarified. We applied an approach that integrated network pharmacology and experimental validation to systemically reveal the molecular mechanism of triptolide in the treatment of MN. Methods: First, potential targets of triptolide and the MN-related targets were collected from publicly available database. Then, based on a protein-protein interaction network as well as GO and KEGG pathway enrichment analyses, we constructed target-pathway networks to unravel therapeutic targets and pathways. Moreover, molecular docking was applied to validate the interactions between the triptolide and hub targets. Finally, we induced passive Heymann nephritis (PHN) rat models and validated the possible molecular mechanisms of triptolide against MN. Results: The network pharmacology results showed that 118 intersected targets were identified for triptolide against MN, including mTOR, STAT3, CASP3, EGFR and AKT1. Based on enrichment analysis, signaling pathways such as PI3K/AKT, MAKP, Ras and Rap1 were involved in triptolide treatment of MN. Furthermore, molecular docking confirmed that triptolide could bind with high affinity to the PIK3R1, AKT1 and mTOR, respectively. Then, in vivo experiments indicated that triptolide can reduce 24 h urine protein (P < 0.01) and protect against renal damage in PHN. Serum albumin level was significantly increased and total cholesterol, triglycerides, and low-density lipoprotein levels were decreased by triptolide (P < 0.05). Compared with PHN group, triptolide treatment regulated the PI3K/AKT/mTOR pathway according to Western blot analyses. Conclusion: Triptolide could exert antiproteinuric and renoprotective effects in PHN. The therapeutic mechanism of triptolide may be associated with the regulation of PI3K/AKT/mTOR signaling pathway. This study demonstrates the pharmacological mechanism of triptolide in the treatment of MN and provides scientific evidence for basic and clinical research.

Projection of fatigue and neuromuscular changes caused by different excises / Projeção da fadiga e as alterações neuromusculares provocadas em distintas atividades físicas / Proyección de la fatiga y los cambios neuromusculares provocados en diferentes actividades físicas

ABSTRACT Introduction: Athletes' muscles can be weakened by fatigue caused by excessive activity. This limitation compromises their functional capacity and professional performance. The competition's performance correlates positively with muscular quality of function. The changes analysis caused by different athletic activities in muscle contraction by noninvasive tensiomyography reflects the functional state of the muscles. Still, no experiments are adapted to verify the fatigue risk level. Objective: Verify the possible relationship between exercise and neuromuscular fatigue using noninvasive tensiomyography. Methods: 90 athletes were randomly selected in weightlifting, badminton, and athletics sports. Maximum radial displacement, contraction, delay, duration, and relaxation time indices were collected. Muscle fatigue detection was based on the empirical mode decomposition modeling method with the Rogers sensitivity fluctuation rate. All values were collected in the rectus femoris muscle before and after the exercises. They were statistically treated and compared (P<0.05). Results: All athletes showed a decline in maximum radial displacement values after exercise. It reveals that their muscles are in a considerable state of tension, especially in the track and field group (from 8.57±3.42mm to 5.43±2.14mm). However, the slightest change in delay time was observed in the weightlifting group (16.21±4.15ms initial versus 18.34±3.27ms final). Conclusion: Through tensiomyography technology, it is possible to obtain a relationship between exercise and neuromuscular fatigue, analyzing the physical activity effects in a noninvasive way. Evidence Level II; Therapeutic Studies - Investigating the result.

RESUMO Introdução: A musculatura do atleta pode ser debilitada com a fadiga ocasionada pelo excesso de atividade. Essa limitação compromete sua capacidade funcional e desempenho profissional. O desempenho nas competições correlaciona-se positivamente com a qualidade da função muscular. A análise das alterações provocadas por distintas atividades atléticas na contração muscular pela tensiomiografia não invasiva reflete o estado funcional dos músculos, porém não há experimentos adaptados para verificar o grau de risco de fadiga. Objetivo: Verificar a possível relação entre exercício e fadiga neuromuscular utilizando tensiomiografia não invasiva. Métodos: 90 atletas foram selecionados aleatoriamente nos esportes de halterofilismo, badminton e atletismo. Foram coletados os índices de deslocamento radial máximo, tempo de contração, tempo de atraso, duração e tempo de relaxamento. A detecção de fadiga muscular foi baseada no método de modelagem da decomposição do modo empírico com o conceito de taxa de flutuação de sensibilidade de Rogers. Os valores dos três grupos foram coletados no músculo reto femoral antes e depois dos exercícios. Foram tratados estatisticamente e comparados(P<0,05). Resultados: Todos os grupos de atletas apresentaram um declínio nos valores de deslocamento radial máximo após o exercício. Isso revela que seus músculos estão em grande estado de tensão, especialmente no grupo de atletismo (de 8.57±3.42mm para 5.43±2.14mm). A menor alteração no tempo de atraso, porém, foi observada no grupo de halterofilismo (16.21±4.15ms iniciais contra 18.34±3.27ms finais). Conclusão: Através da tecnologia de tensiomiografia foi possível obter a relação entre exercício e fadiga neuromuscular analisando os efeitos da atividade física de forma não invasiva. Nível de evidência II; Estudos Terapêuticos - Investigação de Resultados.

RESUMEN Introducción: La musculatura del deportista puede estar debilitada por la fatiga causada por una actividad excesiva. Esta limitación compromete su capacidad funcional y su rendimiento profesional. El rendimiento en las competiciones se correlaciona positivamente con la calidad de la función muscular. El análisis de los cambios provocados por las diferentes actividades deportivas en la contracción muscular mediante la tensiomiografía no invasiva refleja el estado funcional de los músculos, pero no existen experimentos adaptados para verificar el grado de riesgo de fatiga. Objetivo: Verificar la posible relación entre el ejercicio y la fatiga neuromuscular mediante una tensiomografía no invasiva. Métodos: Se seleccionaron al azar 90 atletas de los deportes de halterofilia, bádminton y atletismo. Se recogieron los índices de desplazamiento radial máximo, tiempo de contracción, tiempo de retardo, duración y tiempo de relajación. La detección de la fatiga muscular se basó en el método de modelado de descomposición modal empírica con el concepto de tasa de fluctuación de la sensibilidad propuesto por Rogers. Se recogieron los valores de los tres grupos en el músculo recto femoral antes y después de los ejercicios. Se trataron y compararon estadísticamente (P<0,05). Resultados: Todos los grupos de atletas mostraron un descenso en los valores del desplazamiento radial máximo después del ejercicio. Esto revela que sus músculos están en un gran estado de tensión, especialmente en el grupo de atletismo (de 8,57±3,42mm a 5,43±2,14mm). Sin embargo, el menor cambio en el tiempo de retraso se observó en el grupo de levantamiento de pesas (16,21±4,15ms iniciales frente a 18,34±3,27ms finales). Conclusión: A través de la tecnología de la tensiomografía fue posible obtener la relación entre el ejercicio y la fatiga neuromuscular analizando los efectos de la actividad física de forma no invasiva. Nivel de evidencia II; Estudios terapéuticos - Investigación de resultados.

Mitochondrial Dysfunction and Diabetic Nephropathy: Nontraditional Therapeutic Opportunities.

Diabetic nephropathy (DN) is a progressive microvascular diabetic complication. Growing evidence shows that persistent mitochondrial dysfunction contributes to the progression of renal diseases, including DN, as it alters mitochondrial homeostasis and, in turn, affects normal kidney function. Pharmacological regulation of mitochondrial networking is a promising therapeutic strategy for preventing and restoring renal function in DN. In this review, we have surveyed recent advances in elucidating the mitochondrial networking and signaling pathways in physiological and pathological contexts. Additionally, we have considered the contributions of nontraditional therapy that ameliorate mitochondrial dysfunction and discussed their molecular mechanism, highlighting the potential value of nontraditional therapies, such as herbal medicine and lifestyle interventions, in therapeutic interventions for DN. The generation of new insights using mitochondrial networking will facilitate further investigations on nontraditional therapies for DN.

Ligands and Signaling of Mas-Related G Protein-Coupled Receptor-X2 in Mast Cell Activation.

Mas-related G protein-coupled receptor-X2 (MRGPRX2) is known as a novel receptor to activate mast cells (MCs). MRGPRX2 plays a dual role in promoting MC-dependent host defense and immunomodulation and contributing to the pathogenesis of pseudo-allergic drug reactions, pain, itching, and inflammatory diseases. In this article, we discuss the possible signaling pathways of MCs activation mediated by MRGPRX2 and summarize and classify agonists and inhibitors of MRGPRX2 in MCs activation. MRGPRX2 is a low-affinity and low-selectivity receptor, which allows it to interact with a diverse group of ligands. Diverse MRGPRX2 ligands utilize conserved residues in its transmembrane (TM) domains and carboxyl-terminus Ser/Thr residues to undergo ligand binding and G protein coupling. The coupling likely initiates phosphorylation cascades, induces Ca2+ mobilization, and causes degranulation and generation of cytokines and chemokines via MAPK and NF-κB pathways, resulting in MCs activation. Agonists of MRGPRX2 on MCs are divided into peptides (including antimicrobial peptides, neuropeptides, MC degranulating peptides, peptide hormones) and nonpeptides (including FDA-approved drugs). Inhibitors of MRGPRX2 include non-selective GPCR inhibitors, herbal extracts, small-molecule MRGPRX2 antagonists, and DNA aptamer drugs. Screening and classifying MRGPRX2 ligands and summarizing their signaling pathways would improve our understanding of MRGPRX2-mediated physiological and pathological effects on MCs.

Secondhand cigarette smoke induces increased expression of contractile endothelin receptors in rat coronary arteries via a MEK1/2 sensitive mechanism.

OBJECTIVES: Cigarette smoke, a strong risk factor for cardiovascular diseases, upregulates contractile endothelin (ET) receptors in coronary arteries. The present study examined the effects of second hand cigarette smoke exposure on the contractile endothelin receptors and the role of the MEK1/2 pathway in rat coronary arteries. Design: Rats were exposed to secondhand smoke (SHS) for 8 weeks followed by intraperitoneal injection of a MEK1/2 inhibitor, U0126 daily for another 4 weeks. Contractile responses of isolated coronary arteries were recorded by a sensitive wire myograph. The receptor protein expression levels were examined by Western blotting. Results: The results showed that SHS in vivo caused increased expression of ET receptors ETA and ETB, and that the MEK1/2 blocker U0126 significantly reversed SHS exposure-increased ETA-mediated contractile responses and protein levels. Similar alterations were observed in ETB receptors. U0126 showed dose-dependent effects on SHS-induced response on contractile property and protein levels of the ETB receptor. However, only the higher dose U0126 (15 mg/kg) had inhibitory effects on the ETA receptor. Conclusions: Taken together, our data show that SHS increases contractile ET receptors and MEK1/2 pathway inhibitor offsets SHS exposure-induced ETA and ETB receptor upregulation in rat coronary arteries.

Hydrogen-rich water alleviates cyclosporine A-induced nephrotoxicity via the Keap1/Nrf2 signaling pathway.

Oxidative stress induced by long-term cyclosporine A (CsA) administration is a major cause of chronic nephrotoxicity, which is characterized by tubular atrophy, tubular cell apoptosis, and interstitial fibrosis in the progression of organ transplantation. Although hydrogen-rich water (HRW) has been used to prevent various oxidative stress-related diseases, its underlying mechanisms remain unclear. This study investigated the effects of HRW on CsA-induced nephrotoxicity and its potential mechanisms. After administration of CsA (25 mg/kg/day), rats were treated with or without HRW (12 mL/kg) for 4 weeks. Renal function and vascular activity were investigated. Histological changes in kidney tissues were analyzed using Masson's trichrome and terminal deoxynucleotidyl transferase dUTP nick-end labeling stains. Oxidative stress markers and the activation of the Kelch-like ECH-associated protein 1 (Keap1)/nuclear factor erythroid 2-related factor 2 (Nrf2) signaling pathway were also measured. We found that CsA increased the levels of reactive oxygen species (ROS) and malonaldehyde (MDA), but it reduced glutathione (GSH) and superoxide dismutase (SOD) levels. Such alterations induced vascular dysfunction, tubular atrophy, interstitial fibrosis, and tubular apoptosis. This was evident secondary to an increase in urinary protein, serum creatinine, and blood urea nitrogen, ultimately leading to renal dysfunction. Conversely, HRW decreased levels of ROS and MDA while increasing the activity of GSH and SOD. This was accompanied by an improvement in vascular and renal function. Moreover, HRW significantly decreased the level of Keap1 and increased the expression of Nrf2, NADPH dehydrogenase quinone 1, and heme oxygenase 1. In conclusion, HRW restored the balance of redox status, suppressed oxidative stress damage, and improved kidney function induced by CsA via activation of the Keap1/Nrf2 signaling pathway.

Finding the optimal dose of vitamin K1 to treat vitamin K deficiency and to avoid anaphylactoid reactions.

Vitamin K1 injection induces severe dose-related anaphylactoid reactions and overdose for the treatment of vitamin K deficiency. We aimed to find an optimal and small dose of vitamin K1 injection to treat vitamin K deficiency and avoid anaphylactoid reactions in animal. Rats were administered a vitamin K-deficient diet and gentamicin to establish vitamin K deficiency model. Behaviour tests were performed in beagle dogs to observe anaphylactoid reactions. The results showed an increased protein induced by vitamin K absence or antagonist II (PIVKA-II) levels, a prolonging of prothrombin time (PT) and activated partial thromboplastin time (APTT) and a decrease in vitamin K-dependent coagulation factor (F) II, VII, IX and X activities in the model group. In vitamin K1 0.01 mg/kg group, the liver vitamin K1 levels increased fivefold and the liver vitamin K2 levels increased to the normal amount. Coagulation markers PT, APTT, FVII and FIX activities returned to normal. Both in the 0.1 and 1.0 mg/kg vitamin K1 groups, coagulation functions completely returned to normal. Moreover, the amount of liver vitamin K1 was 40 (0.1 mg/kg) or 100 (1.0 mg/kg) times as in normal. Vitamin K2 was about 4 (0.1 mg/kg) or 5 (1.0 mg/kg) times as the normal amount. There was no obvious anaphylactoid symptom in dogs with the dose of 0.03 mg/kg, which is equivalent to the dose of 0.01 mg/kg in rats. These results demonstrated that a small dose of vitamin K1 is effective to improve vitamin K deficiency and to prevent anaphylactoid reactions, simultaneously.

H2S Prevents Cyclosporine A-Induced Vasomotor Alteration in Rats.

Cyclosporine A (CsA) induces hypertension after transplantation. Hydrogen sulfide (H2S) was found to have hypotensive/vasoprotective effects in the cardiovascular system. The present study aims to investigate the role of H2S on CsA-induced vascular function disorder in rats. Rats were subcutaneously injected with CsA 25 mg/kg for 21 days. Blood pressure was measured by the tail-cuff method. Vasomotion was determined using a sensitive myograph. Western blotting and immunohistochemistry were used to quantify the protein expression of endothelin type A (ETA) receptor and essential MAPK pathway molecules. Vascular superoxide anion production and serum contents of malondialdehyde were determined. The results showed that sodium hydrosulfide (NaHS), a H2S donor, significantly attenuated the increase of blood pressure and contractile responses, and the upregulation of ETA receptor induced by CsA. In addition, NaHS could restore the CsA decreased acetylcholine-induced vasodilatation. Furthermore, NaHS blocked the CsA-induced elevation of reactive oxygen species level, extracellular signal-regulated kinase and p38 MAPK activities. In conclusion, H2S prevents CsA-induced vasomotor dysfunction. H2S attenuates CsA-induced ETA receptor upregulation, which may be associated with MAPK signal pathways. H2S ameliorates endothelial-dependent relaxation, which may be through antioxidant activity.

The expression of bitter taste receptors in mesenteric, cerebral and omental arteries.

AIM: Bitter taste is sensed by the bitter taste receptor (TAS2R), which is mainly expressed in the tongue as well as in extra-oral organs, such as the gastrointestinal tract, respiratory tract, brain, heart and testis. This study aimed to investigate whether TAS2R is expressed in the mesenteric, cerebral and omental arteries. MAIN METHODS: The expression levels of TAS2R mRNA and protein were determined by reverse-transcription polymerase chain reaction and Western blotting, respectively. The location of TAS2R was determined by immunofluorescence imaging. TAS2R agonists were used in a sensitive myograph to study the function of TAS2R in arteries. KEY FINDINGS: The mRNA of rat TAS2Rs, including rTAS2R39, rTAS2R40, rTAS2R108, rTAS2R114, rTAS2R130, rTAS2R137, and rTAS2R140, was expressed in rat mesenteric and cerebral arteries, but rTAS2R114 was not expressed in the cerebral arteries. The mRNA of human TAS2Rs, including hTAS2R3, hTAS2R4, hTAS2R7, hTAS2R10, hTAS2R14, hTAS2R39 and hTAS2R40, was expressed in human omental arteries. The TAS2R7 protein was expressed in rat mesenteric and cerebral arteries, as well as in human omental arteries. Immunofluorescence imaging confirmed that TAS2R7 was located in vascular smooth muscle cells and endothelial cells. The TAS2R agonists, chloroquine and quinine relaxed rat mesenteric arteries and cerebral arteries and human omental arteries in a concentration-dependent manner. SIGNIFICANCE: TAS2R is expressed in the arteries of systemic circulation, including rat mesenteric and cerebral arteries and human omental arteries. This study provides evidence that TAS2R do exist in the arteries and may be involved in the mediation of vessel functions.

Comparison on sensitivity and specificity of serum miR-122 as biomarkers of drug-induced liver injury / 中国药理学与毒理学杂志

OBJECTIVE To make a comparison of sensitivity and specificity between serum miR-122 and traditional biomarkers of drug-induced liver injury. METHODS Acetaminophen (APAP, 1250 mg · kg-1, ig), α-naphthylisothiocyanate (ANIT, 150 mg · kg-1, ig), methionine-choline deficient diet (MCDD, free feeding) and carbon tetrachloride (CCl4, 50%, ip) were used to establish hepatocellular injury, cholestasis, steatosis and fibrosis models, respectively, which were used to evaluate the sensitivity of serum miR-122 as a biomarker of drug-induced liver injury, when compared with the traditional biomarkers. Isoprenaline hydrochloride (IH) and gentamicin (GM) were used to establish myocardial and renal injury models, respectively, which were used to evaluate the specificity of serum miR-122, when compared with the tradi-tional biomarkers. Serum and liver tissues were collected at different time points during the study. Tradi-tional biomarkers such as glutamic-pyruvic transaminase (GPT), glutamic-oxaloacetic transaminase (GOT) and total bilirubin (TBIL) were measured with an automatic biochemistry analyzer. MiR-122 was detected with real- time quantitative PCR. Histopathological examination with HE staining was performed for liver tissues. RESULTS Serum miR-122 increased significantly earlier [miR-122 was signifi-cantly increased (>2 fold) at 1.5 h, 3 h, 2 weeks and 4 weeks after treatment in the four models respec-tively, while no significant increase (<2 fold) was observed for GPT, GOT and TBIL at 6 h, 12 h, 3 weeks and 6 weeks after treatment in the four models respectively] and more signficantly (the maximum fold change for miR-122 was 235.8, 202.7, 73.8 and 600.3 for the four models, respectively. For the GPT, the maximum fold change was 9.5, 3.9, 2.5 and 6.6, respectively; 6.0, 2.4, 1.4 and 3.5 respectively for GOT; 2.6, 2.3, 1.2 and 1.8 respectively for TBIL) than that of traditional biomarkers in hepatocellular injury, cholestasis, steatosis and fibrosis models, when compared with the control group. In the myocar-dial injury model, a significant increase of GOT was observed after IH treatment (2.1 fold), while no change was observed for serum miR-122. In the renal injury model, no false positive results were observed for serum miR-122. CONCLUSION Serum miR-122 can be used as a biomarker of drug-induced liver injury and serum miR-122 is more sensitive and specific than traditional biomarkers (such as GPT, GOT and TBIL).

The effects of MEK1/2 inhibition on cigarette smoke exposure-induced ET receptor upregulation in rat cerebral arteries.

Cigarette smoking, a major stroke risk factor, upregulates endothelin receptors in cerebral arteries. The present study examined the effects of MEK1/2 pathway inhibition on cigarette smoke exposure-induced ET receptor upregulation. Rats were exposed to the secondhand smoke (SHS) for 8weeks followed by intraperitoneal injection of MEK1/2 inhibitor, U0126 for another 4weeks. The urine cotinine levels were assessed with high-performance liquid chromatography. Contractile responses of isolated cerebral arteries were recorded by a sensitive wire myograph. The mRNA and protein expression levels of receptor and MEK/ERK1/2 pathway molecules were examined by real-time PCR and Western blotting, respectively. Cerebral artery receptor localization was determined with immunohistochemistry. The results showed the urine cotinine levels from SHS exposure group were significantly higher than those from the fresh group. In addition, the MEK1/2 inhibitor, U0126 significantly reduced SHS exposure-increased ETA receptor mRNA and protein levels as well as contractile responses mediated by ETA receptors. The immunoreactivity of increased ETA receptor expression was primarily cytoplasmic in smooth muscle cells. In contrast, ETB receptor was noted in endothelial cells. However, the SHS-induced decrease in endothelium-dependent relaxation was unchanged after U0126 treatment. Furthermore, SHS increased the phosphorylation of MEK1/2 and ERK1/2 protein in cerebral arteries. By using U0126 could inhibit the phosphorylated ERK1/2 protein but not MEK1/2. Taken together, our data show that treatment with MEK1/2 pathway inhibitor offsets SHS exposure-induced ETA receptor upregulation in rat cerebral arteries.

Formononetin upregulates nitric oxide synthase in arterial endothelium through estrogen receptors and MAPK pathways.

OBJECTIVES: Formononetin, a phytoestrogen, can improve arterial endothelial cell function by upregulating endothelial nitric oxide synthase (eNOS). The estrogen receptor plays an important role in the regulation of eNOS. This study investigated the hypothesis that formononetin upregulates eNOS through estrogen receptors and MAPK pathways. METHODS: The rat superior mesenteric arteries were cultured with formononetin or formononetin plus inhibitors for 24 h. The isometric tension of the arteries was measured using a myograph system. The mRNA and protein expression levels of eNOS were determined by real-time PCR and immunohistochemistry, respectively. KEY FINDINGS: Acetylcholine (ACh) relaxed the mesenteric arteries precontracted with 5-hydroxytryptamine. This relaxation could be enhanced by formononetin. The removal of endothelium or incubation with l-NAME (a NOS inhibitor) completely abolished the formononetin-enhanced relaxation induced by ACh, suggesting that the formononetin-enhanced vasodilatation is dependent on endothelium and NO pathway. The estrogen receptor inhibitor ICI 182780 attenuated the formononetin-enhanced vasodilatation induced by ACh, suggesting that the formononetin-enhanced arterial relaxation is mediated by the estrogen receptor. Formononetin increased the mRNA and protein expression levels of eNOS. ICI 182780, U0126 (an ERK1/2 inhibitor) and SP600125 (a JNK inhibitor) prevented the increases in arterial relaxation and eNOS levels. CONCLUSIONS: Formononetin upregulates eNOS expression in mesenteric arteries via estrogen receptors, ERK1/2 and JNK pathways.

H2S induces vasoconstriction of rat cerebral arteries via cAMP/adenylyl cyclase pathway.

Hydrogen sulfide (H2S), traditionally known for its toxic effects, is now involved in regulating vascular tone. Here we investigated the vasoconstrictive effect of H2S on cerebral artery and the underlying mechanism. Sodium hydrosulfide (NaHS), a donor of H2S, concentration-dependently induced vasoconstriction on basilar artery, which was enhanced in the presence of isoprenaline, a ß-adrenoceptor agonist or forskolin, an adenylyl cyclase activator. Administration of NaHS attenuated the vasorelaxant effects of isoprenaline or forskolin. Meanwhile, the NaHS-induced vasoconstriction was diminished in the presence of 8B-cAMP, an analog of cAMP, but was not affected by Bay K-8644, a selective L-type Ca(2+) channel agonist. These results could be explained by the revised effects of NaHS on isoprenaline-induced cAMP elevation and forskolin-stimulated adenylyl cyclase activity. Additionally, NaHS-induced vasoconstriction was enhanced by removing the endothelium or in the presence of L-NAME, an inhibitor of nitric oxide synthase. L-NAME only partially attenuated the effect of NaHS which was given together with forskolin on the pre-contracted artery. In conclusion, H2S induces vasoconstriction of cerebral artery via, at least in part, cAMP/adenylyl cyclase pathway.

An optimal initial tension for rat basilar artery in wire myography.

AIMS: The aim of the present study was to determine the optimal initial tension for the rat basilar artery when using wire myography. METHODS: Rat basilar arteries were mounted in myograph baths. A normalization procedure was performed. K(+)-rich (60mM) buffer solution-induced tension was measured in different initial tensions. RESULTS: The initial tension of the basilar artery increased from 0.47 to 2.68mN/mm. Contractile tension was also elevated along with the initial tension. When the initial tension reached 1.63mN/mm, K(+)-induced contractile tension of basilar artery achieved its maximum. Thereafter, contractile tension declined as initial tension increased. The duration of equilibration time did not affect K(+)-induced contractile tension. CONCLUSION: The optimal initial tension is 1.63±0.01mN/mm in rat basilar arteries when using wire myography.

[Correlation between expression of SIL-TAL1 fusion gene and deletion of 6q in T-cell acute lymphoblastic leukemia].

The present study was designed to investigate the prevalence and clinical significance of SIL-TAL1 rearrangements in T-cell acute lymphoblastic leukemia (T-ALL). The incidence of SIL-TAL1 rearrangements was analyzed by nest real-time quantitative polymerase chain reaction (RT-PCR) in 68 patients with T-ALL. Karyotypic analysis was performed by conventional R-banding assay and array-based comparative genomic hybridization (array-CGH). The results showed that SIL-TAL1 rearrangements were identified in 10/26 (38.5%) pediatric and 2/42 (4.8%) adult T-ALL cases, which indicate a pediatric preference for SIL-TAL1 rearrangements in T-ALL. Two different transcripts were detected in 6/12(50%) T-ALL samples. Abnormal karyotypes were detected in 6 out of 11 cases (54.5%) and a deletion of the long arm of chromosome 6 was observed in 4 cases. Array-CGH results of 2 T-ALL cases with SIL-TAL1 rearrangement revealed that this fusion gene was resulted from a cryptic deletion of 1p32, and the overlap region of 6q deletion was 6q14.1-16.3. These cases with SIL-TAL1 fusion had a higher white blood cell (WBC) count and higher serum levels of lactate dehydrogenase (LDH) than cases without SIL-TAL1 fusion. It is concluded that SIL-TAL1 rearrangements are associated with loss of heterozygosity of chromosomal 6q, and SIL-TAL1-positive patients are younger than SIL-TAL1-negative patients. In contrast to the cases without SIL-TAL1 fusion, there are many adverse prognostic factors in the cases with SIL-TAL1 fusion, such as higher WBC count and higher LDH levels.

[Clinical characteristics and prognoses of 167 acute erythroleukemia patients].

OBJECTIVE: To observe the biological characteristic and the prognoses in patients with acute erythroleukemia (AEL). METHODS: The results of 167 patients with newly diagnosed AEL, from January 2003 and June 2013 in the First Affiliated Hospital of Soochow University, were reviewed by MICM. RESULTS: Flow cytometry analysis indicated that CD13(96.1%), CD33(95.1%), CD117(87.4%) and CD34 (79.4%) were highly expressed in AEL. 56 of 148 (37.8%) AEL patients had a variety of cytogenetic abnormalities, 27 of 148(18.2%) patients were complex karyotype (abnormalities involving 3 or more chromosomes), the abnormalities of chromosomes 3, 5, 7 and 8 were more frequently involved and the most common one was ＋8, accounting for 35.7% of all abnormal karyotype, followed by 5q- (17.9%). Mutation analysis showed CEBPA mutation ratio of AEL patients was 44.0% (11/25), that of NPM1 as 15.4% (4/26). Initial induced remission rate of AEL was 56.6% (30/53), compared by 33.3% (4/12) of MDSM6. Survival analysis showed that the overall survival in female was better than that in male (P=0.047). The overall survival time of transplantation group is significantly longer than chemotherapy group (P=0.000). The OS of 13-39 years old group was the best, 40-49 years old group took second place, >50 years old group appeared to be the worst. CONCLUSION: AEL had its own unique biological features, and allogeneic hematopoietic stem cell transplantation could significantly improve its poor prognosis.

The severe adverse reaction to vitamin k1 injection is anaphylactoid reaction but not anaphylaxis.

The severe adverse reaction to vitamin K1 injection is always remarkable and is thought to result from anaphylaxis. Paradoxically, however, some patients administered vitamin K1 injection for the first time have adverse reactions. Using beagle dogs, the present study tested the hypothesis that the response to vitamin K1 is an anaphylactoid reaction. The results showed that serious anaphylaxis-like symptoms appeared in beagle dogs after the administration of vitamin K1 injection for the first time. The plasma histamine concentration increased, and blood pressure decreased sharply. After sensitization, dogs were challenged with vitamin K1 injection and displayed the same degree of symptoms as prior to sensitization. However, when the vitamin K1 injection-sensitized dogs were challenged with a vitamin K1-fat emulsion without solubilizers such asTween-80, the abnormal reactions did not occur. Furthermore, there was no significant change in the plasma immunoglobulin E concentration after vitamin K1 challenge. Following treatment with vitamin K1 injection, the release of histamine and ß-hexosaminidase by rat basophilic leukemia-2H3 cells as well as the rate of apoptosis increased. The Tween-80 group displayed results similar to those observed following vitamin K1 injection in vivo. However, the dogs in the vitamin K1-fat emulsion group did not display any abnormal behavior or significant change in plasma histamine. Additionally, degranulation and apoptosis did not occur in rat basophilic leukemia-2H3 cells. Our results indicate that the adverse reaction induced by vitamin K1 injection is an anaphylactoid reaction, not anaphylaxis. Vitamin K1 injection induces the release of inflammatory factors via a non-IgE-mediated immune pathway, for which the trigger may be the solubilizer.

[Clinical and laboratorial analysis for 15 adult cases of mixed phenotypic acute leukemia with Ph chromosome and/or positive BCR-ABL].

The purpose of this study was to summary the clinical and laboratorial features in 15 adult cases of mixed phenotypic acute leukemia with Ph chromosome and/or BCR-ABL fusion gene positive (Ph(+)MPAL), 15 adult patients with Ph(+)MPAL were defined by WHO-2008 classification. The clinical characteristics, results of morphology, immunology, cytogenetics and molecular genetic detections and results of follow-up in 15 adult patients with Ph(+)MPAL were analyzed retrospectively. The results showed that 15 patients among 87 cases of MPAL demonstrated Ph(+)MPAL (17.2%; 15/87) (7 males and 8 females), their median age was 51 (range 16-81) year old and median WBC count at diagnosis was 69 (12.7-921)×10(9)/L. Based on FAB criteria, these patients showed different morphologic types, including AML (13.3%; 2/15), ALL (40.0%; 6/15), HAL (46.7%; 7/15). Immunologic analysis indicated that 15 cases of Ph(-)MPAL were all classified as B-lymphoid +myeloid mixed immunophenotype. Except one patient, all expressed CD34 antigen on the surface of leukemia cells with 64.3% strong positive, only Ph (53.3%; 8/15), Ph with additional chromosomal abnormalities (33.3%; 5/15) and normal karyotype (13.3%; 2/15) were cytogenetically identified. BCR-ABL fusion gene transcript positive were detected by multiplex reverse transcription PCR in all cases, with e1a2 subtype (p190) (40.0%; 6/15) and b2a2 or b3a2 (p210) subtype (60.0%; 9/15). Four out of 7 (57.1%) patients were found to have IKZF1 gene deletion, without other common gene mutations. Seven out of 10 cases (70.0%) achieved complete remission (CR) after one cycle of induction chemotherapy. In the induction stage, CR rate seemed higher when tyrosine kinase inhibitors (TKI) were added to chemotherapy (83.3%:50.0%; P = 0.206). Overall survival (OS) in 4 patients received allogeneic hematopoietic stem cell transplantation (allo-HSCT) was longer than that in 4 patients received chemotherapy alone (P = 0.004). It is concluded that Ph(+)MPAL mainly is expressed as B+My phenotype. The majority of patients is older and has CD34 overexpression. In the aspect of molecular genetics, the Ph(+)MPAL is similar to other acute leukemia with Ph chromosome. Ph(+)MPAL is a subtype of acute leukemia with poor prognosis. WBC count at diagnosis is an independent prognostic factor. The combination of TKI and allo-HSCT can improve their long-term survival, which needs to be confirmed through carrying out a prospective and multicenter clinical trial for newly diagnosed Ph(+)MPAL.