Abstract Objective: To investigate the optimal timing of initial intravenous immunoglobulin (IVIG) treatment in Kawasaki disease (KD) patients. Methods: KD patients were classified as the early group (day 1-4), conventional group (day 5-7), conventional group (day 8-10), and late group (after day 10). Differences among the groups were analyzed by ANOVA and Chi-square analysis. Predictors of IVIG resistance and the optimal cut-off value were determined by multiple logistic regression analyses and receiver operating characteristic (ROC) curve analysis. Results: There were no significant differences in IVIG resistance among the 4 groups (p = 0.335). The sensitivity analysis also confirmed no difference in the IVIG resistance between those who started the initial IVIG ≤ day 7 of illness and those who received IVIG >day 7 of illness (p = 0.761). In addition, patients who received IVIG administration more than 7 days from the onset had a higher proportion of coronary artery abnormalities (p = 0.034) and longer length of hospitalization (p = 0.033) than those who started IVIG administration less than 7 days. The optimal cut-off value of initial IVIG administration time for predicting IVIG resistance was >7 days, with a sensitivity of 75.25% and specificity of 82.41%. Conclusions: IVIG therapy within 7 days of illness is found to be more effective for reducing the risk of coronary artery abnormalities than those who received IVIG >day 7 of illness. IVIG treatment within the 7 days of illness seems to be the optimal therapeutic window of IVIG. However, further prospective studies with long-term follow-up are required.

Abstract Background Alpha-kinase 1 (ALPK1) is a master regulator in inflammation and has been proved to promote renal fibrosis by promoting the production of IL-1β in diabetic nephropathy (DN) mice. Pyroptosis is involved in high glucose (HG)-induced tubular cells injury, characterized by activation of Gasdermin D (GSDMD) and the release of IL-1β and IL-18, resulting in inflammatory injury in DN. It is reasonable to assume that ALPK1 is involved in pyroptosis-related tubular injury in DN. However, the mechanism remains poorly defined. Methods Immunohistochemistry (IHC) staining was performed to detect the expression of pyroptosis- and fibrosis-related proteins in renal sections of DN patients and DN mice. DN models were induced through injection of streptozotocin combined with a high-fat diet. Protein levels of ALPK1, NF-κB, Caspase-1, GSDMD, IL-1β, IL-18 and α-SMA were detected by Western blot. HK-2 cells treated with high-glucose (HG) served as an in vitro model. ALPK1 small interfering RNA (siRNA) was transfected into HK-2 cells to down-regulate ALPK1. The pyroptosis rates were determined by flow cytometry. The concentrations of IL-1β and IL-18 were evaluated by ELISA kits. Immunofluorescence staining was used to observe translocation of NF-κB and GSDMD. Results The heat map of differentially expressed genes showed that ALPK1, Caspase-1 and GSDMD were upregulated in the DN group. The expression levels of ALPK1, Caspase-1, GSDMD and CD68 were increased in renal biopsy tissues of DN patients by IHC. ALPK1expression and CD68+ macrophages were positively correlated with tubular injury in DN patients. Western blot analysis showed increased expressions of ALPK1, phospho-NF-κB P65, GSDMD-NT, and IL-1β in renal tissues of DN mice and HK-2 cells, accompanied with increased renal fibrosis-related proteins (FN, α-SMA) and macrophages infiltration in interstitial areas. Inhibition of ALPK1 attenuated HG-induced upregulation expressions of NF-κB, pyroptosis-related proteins Caspase-1, GSDMD-NT, IL-1β, IL-18, α-SMA, and pyroptosis level in HK-2 cells. Also, the intensity and nuclear translocation of NF-κB and membranous translocation of GSDMD were ameliorated in HG-treated HK-2 cells after treatment with ALPK1 siRNA. Conclusions Our data suggest that ALPK1/NF-κB pathway initiated canonical caspase-1-GSDMD pyroptosis pathway, resulting in tubular injury and interstitial inflammation of DN.

Abstract Dried radish is a kind of pickled vegetable product with a unique taste made from radish via traditional solid-state spontaneous fermentation. In this study, the effects of variety on physicochemical indexes, texture properties, and taste substances of spontaneous fermented dried radish were investigated. As a result, processing and fermentation had similar trends for all radish varieties. The pH value decreased slightly, soluble solid increased with the decrease of moisture content, the hardness and chewiness decreased, while the content of organic acids and free amino acids increased. Furthermore, the variety of raw material is the key to affecting product quality and taste. The dried radish processed by the variety P21-25 presents the best texture, the highest content of sweet amino acids, the most abundant component of organic acids, while having the lowest salinity. These results indicated that radish varieties could significantly influence the quality and taste of dried radishes under the same processing and fermentation conditions. These findings reveal the importance of variety selection for the quality and taste of spontaneous fermented dried radish, and provide theoretical support for the processing and application of green radish varieties.

Pulmonary fibrosis (PF) is a major public health issue with limited treatment options. As the active ingredient of the n-butanol extract of Amygdalus mongolica (BUT), amygdalin inhibits PF. However, its mechanisms of action are unclear and need further verification. Therefore, the purpose of the present studies was to investigate the anti-fibrotic effects of BUT on PF by serum metabolomics and the transforming growth factor β (TGF-β) pathway. Sixty male Sprague-Dawley rats were randomly divided into control, untreated PF, prednisone-treated (5 mg/kg), and BUT-treated (1.75, 1.25, 0.75 g/kg) groups, and the respective drugs were administered intragastrically for 21 days. The serum metabolomics profiles were determined by ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-QTOF/MS) and metabolism network analysis. The expression of TGF-β1, Smad-3, Smad-7, and α-smooth muscle actin (α-SMA) was measured using a real-time polymerase chain reaction in the lung tissue. BUT significantly alleviated fibrosis by reducing the mRNA expressions of TGF-β1 (from 1.73 to 1.13), Smad-3 (from 2.01 to 1.19), and α-SMA (from 2.14 to 1.19) and increasing that of Smad7 (from 0.17 to 0.62). Twenty-eight potential biomarkers associated with PF were identified. In addition, four key biomarkers were restored to baseline levels following BUT treatment, with the lowest dose showing optimal effect. Furthermore, A. mongolica BUT was found to improve PF by the pentose phosphate pathway and by taurine, hypotaurine, and arachidonic acid metabolism. These findings revealed the mechanism of A. mongolica BUT antifibrotic effects and metabolic activity in PF rats and provided the experimental basis for its clinical application.

Abstract Objectives: Abdominal Aortic Aneurysm (AAA) is a complex disease with both genetic and environmental risk factors. This study aimed to examine the potential association of the +276G/T and −420C>G polymorphisms in the resistin gene with AAA susceptibility and progression. Method: We performed a retrospective study involving AAA patients and healthy controls, assessing the distribution of the +276G/T and −420C>G genotypes in both groups. Hardy-Weinberg equilibrium was assessed for both polymorphisms. Logistic regression was used to explore the influence of these genotypes on AAA occurrence and progression, adjusting for relevant confounders. Results: The distribution of +276G/T polymorphism did not significantly differ between AAA patients and controls. Conversely, a significant difference was observed in the genotype distribution of −420C>G polymorphism between the two groups. The CC genotype and CC/CG genotypes of −420C>G polymorphism were found to be associated with an increased risk and progression of AAA. Conclusions: The −420C>G polymorphism, particularly the CC genotype and CC/CG genotypes, might play a substantial role in AAA susceptibility and progression. The present findings underscore the need for further investigations to confirm these associations and fully elucidate the role of the resistin gene in AAA. Objectives (AAA factors 276GT GT 276G T G 420CG CG 420C C Method controls groups HardyWeinberg Hardy Weinberg confounders Results Conversely CCCG Conclusions

RESUMO Introdução Lesão no menisco do joelho é uma lesão esportiva comum e a cirurgia minimamente invasiva sob artroscopia no joelho tornou-se um método ideal para tratar lesões no menisco. A reabilitação dessa cirurgia vem sendo aprimorada e vários estudos sobre os efeitos do exercício funcional no leque de tratamento ainda são inconclusivos. Objetivo Estudar os efeitos da reabilitação com exercício funcional em pacientes após a lesão esportiva do menisco. Métodos Foram selecionados 20 pacientes com lesão menisco-medial a serem operados, incluindo 8 homens e 12 mulheres. Divididos aleatoriamente em 2 grupos: grupo de treinamento neuromuscular e grupo de treinamento força (11). Sinais e sintomas foram avaliados antes do tratamento e 8 semanas após o tratamento, índices de score JOA e teste de marcha foram comparados, as diferenças do impacto da reabilitação em cada grupo foram avaliadas. Resultados Oito semanas após a reabilitação dos dois grupos, os escores do grupo de treinamento de força foram superiores aos do grupo neuromuscular, a diferença entre os grupos foi estatisticamente significante (P<0,05). Conclusão O exercício funcional acelera a recuperação das articulações, refletida no aumento da força dos músculos adjacentes. O efeito do treinamento muscular e articular na lesão do menisco pós-operatório é digno de reconhecimento. A baropodometria revelou distinções no padrão de marcha entre os diferentes métodos de reabilitação. Na perspectiva desta pesquisa, métodos de reabilitação combinados com exercícios proprioceptivos são complementares. Nível de evidência II; Estudos Terapêuticos - Investigação de Resultados.

ABSTRACT Introduction Knee meniscus injury is a common sports injury, and minimally invasive surgery under knee arthroscopy has become an ideal method to treat meniscus injuries. This surgery rehabilitation has been improved, and several studies on the effects of functional exercise in the range of treatment are still inconclusive. Objective Study the functional exercise rehabilitation effects in patients after sports meniscus injury. Methods Twenty patients with meniscus-medial injury being operated on were selected, including eight men and 12 women. They were randomly divided into neuromuscular and strength training groups (11). Signs and symptoms were assessed before and eight weeks after treatment. JOA score indices and gait tests were compared. The impact of rehabilitation differences was evaluated in each group. Results Eight weeks after rehabilitation in both groups, the scores of the strength training group were higher than the neuromuscular group; the difference between the groups was statistically significant (P<0.05). Conclusion Functional exercise accelerates joint recovery, reflected in increased strength of adjacent muscles. The muscle and joint training effects on postoperative meniscus injury are worthy of recognition. The baropodometry revealed distinctions in walking patterns between different rehabilitation methods. From the perspective of this research, rehabilitation methods combined with proprioceptive exercises are complementary. Evidence Level II; Therapeutic Studies - Investigating the result.

RESUMEN Introducción La lesión de menisco de la rodilla es una lesión deportiva común y la cirugía mínimamente invasiva por artroscopia de rodilla se ha convertido en un método ideal para tratar las lesiones de menisco. La rehabilitación de esta cirugía ha sido mejorada y varios estudios sobre los efectos del ejercicio funcional en el rango de tratamiento aún no son concluyentes. Objetivo Estudiar los efectos de la rehabilitación con ejercicio funcional en pacientes tras una lesión de menisco deportiva. Métodos Se seleccionaron 20 pacientes con lesión de menisco-medial para ser operados, incluyendo 8 hombres y 12 mujeres. Se dividieron aleatoriamente en 2 grupos: grupo de entrenamiento neuromuscular y grupo de entrenamiento de fuerza (11). Se evaluaron los signos y síntomas antes del tratamiento y 8 semanas después del mismo, se compararon los índices de puntuación JOA y la prueba de marcha, y se evaluaron las diferencias del impacto de la rehabilitación en cada grupo. Resultados Ocho semanas después de la rehabilitación para ambos grupos, las puntuaciones del grupo de entrenamiento de fuerza fueron mayores que las del grupo neuromuscular, la diferencia entre los grupos fue estadísticamente significativa (P<0,05). Conclusión El ejercicio funcional acelera la recuperación de la articulación, lo que se refleja en el aumento de la fuerza de los músculos adyacentes. El efecto del entrenamiento muscular y articular en la lesión postoperatoria del menisco es digno de reconocimiento. La baropodometría reveló diferencias en el patrón de la marcha entre los diferentes métodos de rehabilitación. Desde la perspectiva de esta investigación, los métodos de rehabilitación combinados con los ejercicios propioceptivos son complementarios. Nivel de evidencia II; Estudios terapéuticos - Investigación de resultados.

SUMMARY: This study aimed to compare the clinical value of carotid ultrasound and digital subtraction angiography (DSA) for carotid artery stenosis in patients with cerebral infarction. Sixty patients with cerebral infarction underwent carotid ultrasound and DSA. Carotid artery stenosis, degree of stenosis (mild, moderate, severe, and occlusion), and carotid artery plaques were recorded and compared. Carotid stenosis rate was 96.67 % (58/60) and 91.67 % (55/60) on DSA and carotid ultrasound, respectively, and the difference was not statistically significant. Mild, moderate, and severe carotid artery stenosis and occlusion were diagnosed in 35, 28, 20, and 17 arteries, respectively, with DSA, and in 39, 25, 10, and 9 arteries, respectively, with carotid ultrasound. There was a statistically significant difference in the degree of carotid stenosis between the two methods (p<0.05). The kappa value of carotid plaques detected by carotid ultrasound and DSA was 0.776, indicating good consistency. Both carotid ultrasound and DSA are effective for screening carotid artery stenosis and carotid atherosclerotic plaques. While carotid ultrasound is faster and more convenient, DSA can more accurately detect the degree of stenosis and presence of occlusion. Thus, our recommendation is a combination of carotid ultrasound and DSA in clinical settings to improve the convenience and accuracy of diagnosis.

RESUMEN: Este estudio tuvo como objetivo comparar el valor clínico de la ecografía carotídea y la angiografía por sustracción digital (DSA) para la estenosis de la arteria carótida en pacientes con infarto cerebral. Sesenta pacientes con infarto cerebral fueron sometidos a ecografía carotídea y DSA. Se registraron y compararon la estenosis de la arteria carótida, el grado de estenosis (leve, moderada, grave y oclusión) y las placas de la arteria carótida. La tasa de estenosis carotídea fue del 96,67 % (58/60) y del 91,67 % (55/60) en DSA y ecografía carotídea, respectivamente, y la diferencia no fue estadísticamente significativa. Se diagnosticaron estenosis y oclusión de la arteria carótida leve, moderada y grave en 35, 28, 20 y 17 arterias, respectivamente, con DSA, y en 39, 25, 10 y 9 arterias, respectivamente, con ecografía carotídea. Hubo una diferencia estadísticamente significativa en el grado de estenosis carotídea entre los dos métodos (p<0,05). El valor kappa de las placas carotídeas detectadas por ecografía carotídea y DSA fue de 0,776, lo que indica una buena consistencia. Tanto la ecografía carotídea como la DSA son eficaces para detectar la estenosis de la arteria carótida y las placas ateroscleróticas carotídeas. Si bien la ecografía carotídea es más rápida y conveniente, la DSA puede detectar con mayor precisión el grado de estenosis y la presencia de oclusión. Por lo tanto, nuestra recomendación es una combinación de ecografía carotídea y DSA en entornos clínicos para mejorar la conveniencia y precisión del diagnóstico.

ABSTRACT Drought stress and nutrient deficiency are two main factors that restrict the growth and yield of peanut (Arachis hypogaea L.) in arid and semi-arid regions. However, the effects of N fertilizer on root growth and pod yield of peanut under different water conditions are unclear. The growth, root morphology, and pod yield of peanut were studied under different water and N combinations. Two soil water treatments were tested: Well-watered (WW, 80%-85% field capacity) and drought stress conditions (DS, 55%-60% field capacity). Three N treatments were tested: no N application (NN), moderate N (MN, 90 kg ha-1), and high N (HN, 180 kg ha-1). Results showed that pod yield of peanut was limited by drought and N deficiency. Compared with NN, peanut shoot DM and pod yield were improved by MN under DS, with an average increase range of pod yield of 16.2% in the 2 yr. The annual average root length density (RLD) in the deeper soil layer (40-100 cm) was 5.5% higher under DS-MN than DS-NN treatment. The N use efficiency (NUE) was 12.3% higher under WW than DS treatment, and the NUE of MN was significantly higher than that of HN under different water treatments. Root lengths in 40-60 and 60-80 cm soil layers were also positively correlated with pod yield (r = 0.66** and 0.47**, respectively) and NUE (r = 0.60* and 0.85**, respectively). In this study, root growth in deeper soil was improved by MN application under DS, thus increasing yield and NUE of DS conditions.

Abstract Cloudy apple juice (CAJ) is popular due to its potential health benefits and pleasant flavor. However, the common used sterilization methods of CAJ by High hydrostatic pressure (HHP) or high-temperature short time (HTST) still showed many inadequacies. The present work studied the impact of HHP (400MPa/10 °C/10 min) and HTST (98 °C/50s) on CAJs made from a new cultivar Changfu 2 planted in Loess plateau area. We comparatively evaluated the microbes inactivation, enzymatic activity, phytochemical parameters, and sensory quality after the HHP and HTST treatment and during the cold storage (4 °C). Results showed that during the cold storage, both HHP and HTST treated CAJ showed a declining trend in sensory attributes like aroma, taste and overall acceptability. HTST showed a higher killing rate of bacteria, and the CAJ was detected a higher diversity of microorganisms (expressed as OTU) during the cold storage. At the end of shelf-life, Rickettsia, oxyphotobacteria, and pseudomonas were detected as the dominant genera in both HHP and HTST treated CAJs. Spearman bivariate correlate analysis determined the relationship between the bacterial profiles and enzymatic activity, phytochemical parameters, and sensory quality after the HHP and HTST treatment.

The fluorescent microspheres are prepared by the swelling-evaporation method. The optimal swelling conditions are obtained by researching related factors, such as types and dosages of swelling agent, dosages of fluorescent substance, swelling time and swelling temperature. Considering that the structure of microspheres can influence the swelling properties, the paper mainly focuses on the effect of molecular weight and surface carboxyl group content of polystyrene microspheres on swelling properties. When the molecular weight of microspheres is 75276 g mol-1, the density of carboxyl group on their surface is 6.94×10-4 mol g-1, and the doping amount of europium complex reaches the maximum of 24.9 mg g-1 with the strongest fluorescence intensity. Meanwhile, the as-prepared fluorescent microspheres have high monodispersity, uniform particles size and high thermal stability (the decomposition temperature is 385oC), which sets up the good foundation for biomolecule detection. So, this work provides a significant experience for the preparation of fluorescent microspheres.

Abstract Ganoderic acid A (GAA) is one of the most abundant triterpenoids in Ganoderma lucidum and has protective effect on several vascular diseases. However, the effect of GAA on pulmonary arterial hypertension (PAH) has not been reported. The aim of this study was to investigate the effect of the GAA on the hypoxia-induced phenotypic modulation of PASMCs and the involved transduction pathway. Primary rat pulmonary artery smooth muscle cells (PASMCs) were isolated and cultured under hypoxia condition to induce phenotypic modulation. Our results showed that hypoxia significantly increased the proliferation and migration of PASMCs, as well as inhibited the apoptosis of PASMCs, which were blocked by GAA treatment. In addition, hypoxia-induced dedifferentiation of PASMCs was prevented by GAA with increased the expression levels of myocardin and calponin, and decreased the expression of osteopontin (OPN). Furthermore, GAA suppressed the hypoxia-induced expression of p-PI3K and p-Akt in PASMCs. Treatment with IGF-1 reversed the effects of GAA on proliferation, migration, apoptosis and dedifferentiation in hypoxia-treated PASMCs. Taken together, these findings demonstrated that GAA suppresses the phenotypic modulation of PASMCs through the inactivation of PI3K/Akt pathway. Thus, GAA may be a potent therapeutic agent for PAH in future clinical practice.

Abstract The aim of this study was to determine the resistance mechanism of Pseudomonas aeruginosa to cefoperazone sodium/sulbactam sodium. We retrospectively analyzed the drug resistance of P.a isolated at the First Affiliated Hospital of Guangxi Medical University. Drug-resistant P.a strains were constructed, then wild-type (WT) and drug-resistant (DR) strains were compared using protein and gene microarrays to determine differences between DR and WT strains. The resistance rates of P. aeruginosa during 2013, 2014 and 2015 were 21.2%, 21.4%, and 24.6% respectively. Among 242 protein peaks of WT and DR bacteriophage proteins, 41 were differentially expressed between the two groups. The expression of 26 and 15 proteins were respectively upregulated and downregulated in the DR compared with the WT group. Gene microarray results revealed 679 mutant loci in the DR group, of which 42 with the top 50 Q values were found in the NCBI database. The rate of P.a resistance to cefoperazone sodium/sulbactam sodium remained high between 2013 and 2015. The numbers of different proteins and genetic variations in the DR strains suggested that the resistance mechanism of P.a to cefoperazone sodium/sulbactam sodium involves multiple genes and proteins that might be key to controlling P.a resistance to cefoperazone sodium/sulbactam sodium.

ABSTRACT Purpose: To investigate the role of renal denervation (RDN) on endogenous ouabain (EO) secretion in spontaneously hypertensive rats (SHR). Methods: Sixteen 12-week-old male SHR were randomly separated into the renal denervation group (RDNX group) and sham operation group (sham group), and eight age-matched Wistar Kyoto rats (WKY) were served as control group. EO concentrations, the Na+- K+-ATPaseactivity, and the expression of Na+-K+-ATPase were assessed. Results: EO levels in serum, kidneys and hypothalamus of sham group were higher than in RDNX group (p < 0.05). Renal Na+-K+-ATPase activity subjected to denervation surgery showed significantly reduction when compared with the sham groups (p < 0.05). A positive correlation existed between norepinephrine (NE) content and Na+-K+-ATPase activity in the kidney (r2 = 0.579). Renal Na+-K+-ATPase α1 subunit mRNA expression was down-regulated in the RDNX group compared with the sham group (P < 0.05), while renal Na+-K+-ATPase α1 subunit mRNA expression was no statistical significance between the groups (P = 0.63). Immunohistochemical analysis showed that there were significant differences in the renal expression of Na+-K+-ATPasebetween the three groups (P < 0.05). Conclusions: These experiments demonstrate that RDN exerted an anti-hypertensive effect with reduction of EO levels and Na+-K+-ATPase activity and Na+-K+-ATPase α1 subunit expression of kidney in SHR.

Abstract Objective: To investigate the impact of recombinant human interferon α1b (rhIFNα1b) treatment in infants hospitalized with lower respiratory tract infections on subsequent wheezing. Methods: The clinical data of infants (n = 540) with viral pneumonia, wheezy bronchitis, or bronchiolitis hospitalized in 19 Chinese hospitals from June 2009 to June 2015 were retrospectively analyzed. The parameters relevant to wheezing episodes within the last year were collected by telephone and questionnaires. The rhIFNα1b treatment group (n = 253) and control group (n = 287) were compared in terms of wheezing episodes within the last year. Moreover, the wheezing group (95 cases) and non-wheezing group (445 cases) were compared. Results: Out of 540 cases, 95 (17.6%) experienced wheezing episodes, 13.8% (35/253) cases treated with rhIFNα1b, and 20.9% (60/287) cases without rhIFNα1b experienced wheezing episodes within the last year. The rhIFNα1b treatment significantly improved wheezing episodes within the last year, compared with the control peers (p = 0.031). Single-factor regression showed statistically significant differences between the wheezing and non-wheezing groups in terms of age, rhIFNα1b use, childhood and family history of allergy, housing situation, and feeding history (p < 0.05). Binary logistic regression showed a childhood history of allergy (OR = 2.14, p = 0.004), no rhIFNα1b use (OR = 1.70, p = 0.028), and living in a crowded house (OR = 1.92, p = 0.012) might be risk factors of subsequent wheezing. Accordingly, breastfeeding (OR = 0.44, p = 0.008) and hospitalization age of 1-year-old (OR = 0.58, p = 0.024) were protective factors. Conclusions: Early use of rhIFNα1b in infants hospitalized with lower respiratory tract infections and breastfeeding could prevent subsequent wheezing. Living in a crowded house could promote subsequent wheezing.

Resumo Fundamento: A doença cardiovascular é a principal causa de morte em todo o mundo. A apoptose mediada por hipóxia em cardiomiócitos é uma das principais causas de distúrbios cardiovasculares. O tratamento com a proteína do fator de crescimento endotelial vascular (VEGF, do inglês vascular endothelial growth factor) foi testado, mas as dificuldades operacionais limitaram seu uso. Entretanto, com os avanços da terapia gênica, aumentou o interesse na terapia gênica baseada no VEGF em doenças cardiovasculares. No entanto, o mecanismo preciso pelo qual a reposição de VEGF resgata os danos pós-hipóxia em cardiomiócitos não é conhecido. Objetivos: Investigar o efeito da expressão de VEGF121 pós-hipóxia utilizando cardiomiócitos de ratos neonatos. Métodos: Cardiomiócitos isolados de ratos neonatos foram utilizados para estabelecer um modelo in vitro de lesão cardíaca induzida por hipóxia. O efeito da superexpressão de VEGF, isolado ou em conjunto com inibidores de moléculas pequenas que têm como alvo os canais de cálcio, receptores sensíveis ao cálcio (CaSR, do inglês calcium-sensitive receptors) e calpaína, no crescimento e proliferação celular em lesão de cardiomiócitos induzidos por hipóxia, foram determinados com ensaio de MTT, coloração TUNEL, coloração com Anexina V/PI, lactato desidrogenase e atividade da caspase. Para análise estatística, um valor de p<0,05 foi considerado significativo. Resultados: Verificou-se que o efeito do VEGF121 foi mediado por CaSR e calpaína, mas não foi dependente dos canais de cálcio. Conclusões: Nossos resultados, mesmo em um ambiente in vitro, estabelecem as bases para uma validação futura e testes pré-clínicos da terapia gênica baseada em VEGF em doenças cardiovasculares.

Abstract Background: Cardiovascular disease is the major cause of death worldwide. Hypoxia-mediated apoptosis in cardiomyocytes is a major cause of cardiovascular disorders. Treatment with vascular endothelial growth factor (VEGF) protein has been tested but operational difficulties have limited its use. However, with the advancements of gene therapy, interest has risen in VEGF-based gene therapy in cardiovascular disorders. However, the precise mechanism by which VEGF replenishment rescues post-hypoxia damage in cardiomyocytes is not known. Objectives: To investigate the effect of post-hypoxia VEGF121 expression using neonatal rat cardiomyocytes. Methods: Cardiomyocytes isolated from neonatal rats were used to establish an in vitro model of hypoxia-induced cardiac injury. The effect of VEGF overexpression, alone or in combination with small-molecule inhibitors targeting calcium channel, calcium sensitive receptors (CaSR), and calpain on cell growth and proliferation on hypoxia-induced cardiomyocyte injury were determined using an MTT assay, TUNEL staining, Annexin V/PI staining, lactate dehydrogenase and caspase activity. For statistical analysis, a value of P<0.05 was considered to be significant. Results: The effect of VEGF121 was found to be mediated by CaSR and calpain but was not dependent on calcium channels. Conclusions: Our findings, even though using an in vitro setting, lay the foundation for future validation and pre-clinical testing of VEGF-based gene therapy in cardiovascular diseases.