Uncovering the Penile Clock: Expression of Molecular Clock Proteins in Human Penile Cavernous Tissue.

PURPOSE: To evaluate the expression of core molecular clock genes/proteins in penile cavernous tissue from healthy male subjects and to determine whether their expression has circadian variation. MATERIALS AND METHODS: Corpus cavernosum biopsy samples were obtained from 10 healthy males with penile deviation or fracture who underwent surgical intervention during the day and night. The daytime group (n=5) underwent corpus cavernosum tissue sampling during zeitgeber time (ZT) 8-12, while the nighttime group (n=5) underwent sampling during ZT 20-24. The expression and localization of BMAL1, CLOCK, PER1, PER2, PER3, CRY1, and CRY2 proteins were analyzed using immunohistochemistry and quantified using H-score analysis. RT-qPCR analysis was performed to assess the expression of core molecular clock genes in the corpus cavernosum tissue of 5 additional daytime patients. RESULTS: The expression of core molecular clock proteins was detected in vascular endothelial cells (VECs) and smooth muscle cells (SMCs) in corpus cavernosum during daytime and nighttime. BMAL1 exhibited the most significant nuclear expression during daytime in both cell types, whereas its expression decreased significantly at night. In VECs, a significant decrease in the nuclear expression of CRY1 was observed at night. In SMCs, a significant decrease in the cytoplasmic expression of PER3 was observed at night. The expression patterns of the core molecular clock genes were ascertained through a RT-qPCR analysis. CONCLUSIONS: Our research provides compelling evidence that core molecular clock genes are distinctly expressed in penile tissue in humans. Furthermore, we observed the expression of molecular clock proteins within the VECs and SMCs of the corpus cavernosum, with BMAL1 being the most prominently expressed. The discovery of core molecular clock genes in penile tissue, as well as proteins within the SMCs and VECs of the corpus cavernosum, introduces the potential significance of the molecular clock mechanism in the physiology of penile erection.

Effects of photobiomodulation therapy (PBMT) over endothelial function in healthy individuals: a preliminary crossover clinical trial.

Photobiomodulation therapy (PBMT) causes stimulatory effects that raise cell metabolism. The study aimed to evaluate the effects of PBMT on the endothelial function of healthy individuals. It was a controlled, randomized, crossover, triple-blind trial with 22 healthy volunteers (female: 77.3%), aged 25.45 years which were randomly divided into three groups. PBMT with gallium-aluminum-arsenide (GaAlAs) diode laser (810 nm, continuous-wave mode, 1000 mW, 0.28 cm2) was applied over the radial and ulnar artery regions in two parallel spots: group 1-30 J (n = 22, 107 J/cm2) per spot; group 2-60 J (n = 22, 214 J/cm2) per spot; and group 3-placebo (n = 22, sham). The endothelial function was measured before and immediately after PBMT by the flow-mediated dilation technique (%FMD) with high-resolution ultrasound. Statistical analysis was made with ANOVA for repeated measures, the effect size was measured by Cohen's d, and results are presented as mean and standard error (or 95% confidence intervals). A p-value < 0.05 was considered statistically significant. The %FMD increases 10.4% with 60 J (mean difference = 0.496 mm, 95% CI = 0.42 to 0.57, p < 0.001), 7.3% with 30 J (mean difference = 0.518 mm, 95% CI = 0.44 to 0.59, p < 0.001), and 4.7% with placebo (mean difference = 0.560 mm, 95% CI = 0.48 to 0.63, p < 0.001). We found a small effect size (p = 0.702; d de Cohen = 0.24) without statistical difference between interventions. PBMT with the energy density of 60 J and 30 J did not improve endothelial function.Trial registration number: NCT03252184 (01/09/2017).

Role of homocysteinylation in cerebral atherosclerosis / 国际脑血管病杂志

Homocysteine is closely associated with extracranial and intracranial atherosclerosis, and its main pathogenesis includes oxidative stress, lipid metabolism disorder and vascular endothelial dysfunction. As a protein modification related to homocysteine, homocysteinylation can promote the occurrence and development of cerebral atherosclerosis by promoting oxidation, changing lipid function and destroying vascular endothelial function. This article reviews the role of homocysteinylation in cerebral atherosclerosis, and discusses the possibility of preventing cerebral atherosclerosis by homocysteinylation.

Pathogenesis of cerebral small vessel disease / 国际脑血管病杂志

Cerebral small vessel disease (CSVD) is an important cause of ischemic stroke and vascular dementia, which brings heavy burden to families and society. The prevention and treatment of CSVD has always been a research hotspot, but its pathogenesis is still not completely clear. This article reviews the pathogenesis of CSVD, including chronic cerebral hypoperfusion, blood-brain barrier dysfunction, vascular endothelial dysfunction, interstitial fluid reflux disorder, inflammatory response, and genetic factors, in order to provide more sufficient theoretical basis for early intervention and treatment of CSVD.

Efficacy and safety of three days continuous intravenous pumping of recombinant human endostatin combined with platium-contained chemotherapeutic agents in patients with advanced non-small cell lung cancer aged 60 years or older / 中华老年医学杂志

Objective:To investigate the efficacy and safety of recombinant human endostatin(Endostar)combined with platium-contained chemotherapeutic agents in advanced non-small cell lung cancer(NSCLC)patients over 60 years old.Methods:93 advanced NSCLC patients from January 2019 to June 2021 were selected as the research objects.The patients received three days of continuous intravenous infusion of Endostar(210 mg for 72 hours)combined with platinum-containing chemotherapy.The efficacy and toxicities were evaluated according to Response Evaluation Criteria in Solid Tumors(RECIST)version1.1 and National Cancer Institute Common Terminology Criteria for Adverse Events(NCI-CTCAE version 4.0), respectively.Follow-up data were obtained to perform the Kaplan-Meier survival analysis.Results:In our study, the objective remission rate(ORR)and disease control rate(DCR)were 38.7% and 78.5%, respectively.The median progression-free survival(PFS)and overall survival(OS)were 6.8 months and 16.5 months, respectively.A Multivariate analysis showed that tumor staging and TP53 mutation were independent prognostic factors related to PFS and OS in advanced NSCLC patients.Adverse reactions related to Endostar during treatment included arrhythmia in 2 cases(2.2%), myocardial ischemia in 1 case(1.07%)and bloody sputum in 1 case(1.07%), all of which were Grade 1 or Grade 2.Conclusions:The application of three days continuous intravenous infusion of Endostar combined with platium-contained chemotherapeutic agents is worthy to be recommended for clinical application in elderly patients with advanced NSCLC due to its high effective rate and survival advantage, as well as good safety.

Plasma Cell-Free DNA Predicts Survival and Maps Specific Sources of Injury in Pulmonary Arterial Hypertension.

BACKGROUND: Cell-free DNA (cfDNA) is a noninvasive marker of cellular injury. Its significance in pulmonary arterial hypertension (PAH) is unknown. METHODS: Plasma cfDNA was measured in 2 PAH cohorts (A, n=48; B, n=161) and controls (n=48). Data were collected for REVEAL 2.0 (Registry to Evaluate Early and Long-Term PAH Disease Management) scores and outcome determinations. Patients were divided into the following REVEAL risk groups: low (≤6), medium (7-8), and high (≥9). Total cfDNA concentrations were compared among controls and PAH risk groups by 1-way analysis of variance. Log-rank tests compared survival between cfDNA tertiles and REVEAL risk groups. Areas under the receiver operating characteristic curve were estimated from logistic regression models. A sample subset from cohort B (n=96) and controls (n=16) underwent bisulfite sequencing followed by a deconvolution algorithm to map cell-specific cfDNA methylation patterns, with concentrations compared using t tests. RESULTS: In cohort A, median (interquartile range) age was 62 years (47-71), with 75% female, and median (interquartile range) REVEAL 2.0 was 6 (4-9). In cohort B, median (interquartile range) age was 59 years (49-71), with 69% female, and median (interquartile range) REVEAL 2.0 was 7 (6-9). In both cohorts, cfDNA concentrations differed among patients with PAH of varying REVEAL risk and controls (analysis of variance P≤0.002) and were greater in the high-risk compared with the low-risk category (P≤0.002). In cohort B, death or lung transplant occurred in 14 of 54, 23 of 53, and 35 of 54 patients in the lowest, middle, and highest cfDNA tertiles, respectively. cfDNA levels stratified as tertiles (log-rank: P=0.0001) and REVEAL risk groups (log-rank: P<0.0001) each predicted transplant-free survival. The addition of cfDNA to REVEAL improved discrimination (area under the receiver operating characteristic curve, 0.72-0.78; P=0.02). Compared with controls, methylation analysis in patients with PAH revealed increased cfDNA originating from erythrocyte progenitors, neutrophils, monocytes, adipocytes, natural killer cells, vascular endothelium, and cardiac myocytes (Bonferroni adjusted P<0.05). cfDNA concentrations derived from erythrocyte progenitor cells, cardiac myocytes, and vascular endothelium were greater in patients with PAH with high-risk versus low-risk REVEAL scores (P≤0.02). CONCLUSIONS: Circulating cfDNA is elevated in patients with PAH, correlates with disease severity, and predicts worse survival. Results from cfDNA methylation analyses in patients with PAH are consistent with prevailing paradigms of disease pathogenesis.

Avaliação da Disfunção Endotelial em Casos de COVID-19 com Dilatação Fluxo-Mediada / Evaluation of Endothelial Dysfunction in COVID-19 With Flow-Mediated Dilatation

Resumo Fundamento: Sabe-se que a inflamação desempenha um papel crucial em muitas doenças, incluindo a COVID-19. Objetivo: Utilizando a dilatação fluxo-mediada (DFM), objetivou-se avaliar os efeitos da inflamação na função endotelial de pacientes com COVID-19. Métodos: Este estudo foi realizado com um total de 161 indivíduos, dos quais 80 foram diagnosticados com COVID-19 nos últimos seis meses (48 mulheres e 32 homens com idade média de 32,10±5,87 anos) e 81 eram controles saudáveis (45 mulheres e 36 homens com idade média de 30,51±7,33 anos). Os achados do ecocardiograma transtorácico e da DFM foram analisados em todos os indivíduos. Resultados com p<0,05 foram considerados estatisticamente significantes. Resultados: O ecocardiograma e a DFM do grupo COVID-19 foram realizados 35 dias (intervalo: 25-178) após o diagnóstico. Não houve diferença estatisticamente significativa nos parâmetros ecocardiográficos. Em contraste, a DFM (%) foi significativamente maior no grupo controle (9,52±5,98 versus 12,01±6,18; p=0,01). Na análise multivariada com o modelo stepwise progressivo, a DFM foi significativamente diferente no grupo controle em relação ao grupo COVID-19 (1,086 (1,026-1,149), p=0,04). O teste de correlação de Spearman indicou que a DFM (r=0,27; p=0,006) apresentou correlação positiva fraca com a presença de COVID-19. Conclusão: Os achados deste estudo apontam para disfunção endotelial induzida por COVID-19, avaliada por DFM, na fase inicial de recuperação.

Abstract Background: Inflammation is known to play a crucial role in many diseases, including COVID-19. Objective: Using flow-mediated dilatation (FMD), we aimed to assess the effects of inflammation on endothelial function in COVID-19 patients. Methods: This study was conducted with a total of 161 subjects, of whom 80 were diagnosed with COVID-19 within the last six months (comprising 48 women and 32 men with a mean age of 32.10 ± 5.87 years) and 81 were healthy controls (comprising 45 women and 36 men with a mean age of 30.51 ± 7.33 years). We analyzed the findings of transthoracic echocardiography and FMD in all subjects. All results were considered statistically significant at the level of p < 0.05. Results: The echocardiography and FMD of the COVID-19 group were performed 35 days (range: 25-178) after diagnosis. There was no statistically significant difference in echocardiographic parameters. Differently, FMD (%) was significantly higher in the control group (9.52 ± 5.98 vs. 12.01 ± 6.18, p=0.01). In multivariate analysis with the forward stepwise model, FMD was significantly different in the control group compared to the COVID-19 group (1.086 (1.026 - 1.149), p=0.04). A Spearman's correlation test indicated that FMD (r=0.27, p=0.006) had a weak positive correlation with the presence of COVID-19. Conclusion: Our findings point to COVID-19-induced endothelial dysfunction, as assessed by FMD, in the early recovery phase.

Glucose Metabolic Disorders Enhance Vascular Dysfunction Triggered by Particulate Air Pollution: a Panel Study.

BACKGROUND: Vascular dysfunction is a biological pathway whereby particulate matter (PM) exerts deleterious cardiovascular effects. The effects of ambient PM on vascular function in prediabetic individuals are unclear. METHODS: A panel study recruited 112 Beijing residents with and without prediabetes. Multiple vascular function indices were measured up to 7 times. The associations between vascular function indices and short-term exposure to ambient PM, including fine particulate matter (PM2.5), ultrafine particles, accumulation mode particles, and black carbon, and the modification of these associations by glucose metabolic status were examined using linear mixed-effects models. RESULTS: Increases in brachial artery pulse pressure, central aortic pulse pressure, and ejection duration, and decreases in subendocardial viability ratio and reactive hyperemia index were significantly associated with at least one PM pollutant in all participants, indicating increased vascular dysfunction. For example, for an interquartile range increment in 5-day moving average ultrafine particles, brachial artery pulse pressure, and central aortic pulse pressure increased 5.4% (0.8%-10.4%) and 6.2% (1.2%-11.5%), respectively. Additionally, PM-associated changes in vascular function differed according to glucose metabolic status. Among participants with high fasting blood glucose levels (≥6.1 mmol/L), PM exposure was significantly associated with increased brachial artery systolic blood pressure, central aortic systolic blood pressure, brachial artery pulse pressure, central aortic pulse pressure, and augmentation pressure normalized to a heart rate of 75 bpm and decreased subendocardial viability ratio and reactive hyperemia index. Weaker or null associations were observed in the low-fasting blood glucose group. CONCLUSIONS: Glucose metabolic disorders may exacerbate vascular dysfunction associated with short-term ambient PM exposure.

Effect of bone morphogenetic protein 4 on glycolysis of human retinal vascular endothelial cells / 中华眼底病杂志

Objective:To explore the effect of bone morphogenetic protein 4 (BMP4) on the glycolysis level of human retinal microvascular endothelial cells (hRMECs).Methods:A experimental study. hRMECs cultured in vitro were divided into normal group, 4-hydroxynonenal (HNE) group (4-HNE group) and 4-HNE+BMP4 treatment group (BMP4 group). 4-HNE group cell culture medium was added with 10 μmmol/L 4-HNE; BMP4 group cell culture medium was added with recombinant human BMP4 100 ng/ml after 6 h stimulation with 10 μmol/L 4-HNE. The levels of intracellular reactive oxygen species (ROS) were detected by flow cytometry. The effect of 4-HNE on the viability of cells was detected by thiazole blue colorimetry. Cell scratch test and Transwell cell method were used to determine the effect of 4-HNE on cell migration. The relative expression of BMP4 and SMAD9 mRNA and protein in normal group and 4-HNE group were detected by realtime quantitative polymerase chain reaction and Western blot. Seahorse XFe96 cell energy metabolism analyzer was used to determine the level of intracellular glycolysis metabolism in normal group, 4-HNE group and BMP4 group. One-way analysis of variance was used for comparison between groups.Results:The ROS levels in hRMECs of normal group, 4-HNE group and BMP4 group were 21±1, 815±5, 810±7, respectively. Compared with the normal group, the levels of ROS in the 4-HNE group and the BMP4 group were significantly increased, and the difference was statistically significant ( F=53.40, 50.30; P<0.001). The cell viability in the normal group and 4-HNE group was 1.05±0.05 and 1.28±0.05, respectively; the migration rates were (0.148±0.005)%, (0.376±0.015)%; the number of cells passing through the pores were 109.0±9.6, 318.0±6.4, respectively. Compared with the normal group, the 4-HNE group had significantly higher cell viability, cell migration rate, and the number of cells passing through the pores, and the differences were statistically significant ( F=54.35, 52.84, 84.35; P<0.05). The relative expression levels of BMP4 and SMAD9 mRNA in the cells of the 4-HEN group were 1.680±0.039 and 1.760±0.011, respectively; compared with the normal group, the difference was statistically significant ( F=53.66, 83.54; P<0.05). The relative expression levels of BMP4 and SMAD9 proteins in the cells of the normal group and 4-HEN group were 0.620±0.045, 0.860±0.190, 0.166±0.049, 0.309±0.038, respectively; compared with the normal group, the differences were statistically significant ( F=24.87, 53.84; P<0.05). The levels of intracellular glycolysis, glycolytic capacity and glycolytic reserve in normal group, 4-HNE group and BMP4 group were 1.21±0.12, 2.84±0.24, 1.78±0.36, 2.59±0.11, 5.34±0.32, 2.78±0.45 and 2.64±0.13, 5.20±0.28, 2.66±0.33. Compared with the normal group, the differences were statistically significant (4-HNE group: F=86.34, 69.75, 58.45; P<0.001; BMP4 group: F=56.87, 59.35, 58.35; P<0.05). There was no significant difference in intracellular glycolysis, glycolysis capacity and glycolysis reserve level between 4-HNE group and BMP4 group ( F=48.32, 56.33, 55.01; P>0.05). Conclusion:BMP4 induces the proliferation and migration of hRMECs through glycolysis.

Effect of catheterin-related antimicrobial peptides on the high glucose induced cardiac microvascular endothelial cell injury / 中华老年医学杂志

Objective:To investigate the effect of catheterin-related antimicrobial peptides(CRAMP)on the damage of cardiac microvascular endothelial cells induced by high glucose.Methods:Adult mouse heart microvascular endothelial cells were isolated and cultured.A model of microvascular endothelial cell injury was established by high glucose culture.The endothelial cells were randomly divided into 4 groups as the following.In the control group, 27.5 mmol/L mannitol was given as isoosmotic control as compared with the high glucose group.In the high glucose group(HG group), cells were cultured with 33.3 mmol/L high glucose for 48 h, and then treated without CRAMP.In 0.15 mg/L CRAMP treatment group, cells were cultured with 33.3 mmol/L high glucose for 48 h, followed by 0.15 mg/L CRAMP treatmen for 24 h. In the 0.5 mg/L CRAMP treatment group, cells were cultured with 33.3 mmol/L high glucose treatment for 48 h, and then treated with 0.5 mg/L CRAMP for 24 h. Cell proliferation was examined by staining with CKK-8 cell counting kit.The secretion of inflammatory factors in microvascular endothelial cells was detected by ELISA kit.Reactive oxygen species assay kit detects the level of reactive oxygen species in cells.Cell apoptosis was detected by apoptosis kit.Tubule formation and tubule number were measured by cells cultured on the matrix glue membrane, then detected by microscopic observation.The nitric oxide(NO)test kit measures levels of NO.The expression of nitric oxide synthase(eNOS)was detected by western blotting.Results:The cell proliferation activity was significant lower in the HG group than in control group[(52.2±5.4)% vs.(100.0±7.3)%]. The cell proliferation activity was higher in the 0.15 and 0.5 mg/L CRAMP groups than in the HG group[(72.0±3.4)% vs.(52.2±5.4)%; and(84.2±5.8)% vs.(52.2±5.4)%( F=75.300, P<0.001)]. The expression of tumor necrosis factor-α was significantly higher in the HG group than in the control group and in 0.5 mg/L CRAMP group[HG group of(239.1±32.1)μg/L, the control of(22.1±3.7)μg/L, 0.5 mg/L CRAMP of(84.6±9.4)μg/L]( F=197.300, P<0.001). The level of reactive oxygen species was significantly higher in the HG group than in control group and in 0.5 mg/L CRAMP group[(20.8±2.4)in HG group, (4.8±1.7)in control group, (10.2±1.5)in CRAMP group]( F=105.700, P<0.001). The number of apoptotic cells was significantly higher in the HG group than in control group and in 0.5 mg/L CRAMP group[(21.2±3.1)% in HG group, (2.2±0.6)% in control group(9.5±1.2)% in CRAMP group]( F=141.900, P<0.001). The length and number of tubules were lower in the HG group than in control group and in CRAMP group[for the length: (87.8±9.1)μm in HG group, (337.0±37.2)μm in control group(206.5±16.3)μm in CRAMP group( F=160.800, P<0.001); for the number: (9.1±1.9)in HG group, (22.0±3.4)in control group, (16.8±2.2)]in CRAMP group( F=36.200, P<0.001)]. The level of NO was lower in the HG group than in control group and in CRAMP group[(0.25±0.05)in HG group, (1.05±0.16)in control group, (0.75±0.06)in CRAMP group( F=83.200, P<0.001)]. The protein expression and mRNA levels of endothelial nitric oxide synthase(eNOS)were lower in the HG group than in the control group and in CRAMP group[for eNOS protein: (0.07±0.03)in HG group, (0.81±0.05)in control group, (0.54±0.07)in CRAMP group, F=275.700, P<0.001; and for eNOS mRNA: (0.11±0.07)in HG group, (1.00±0.22)in control group, (0.57±0.12)in CRAMP group, F=50.600, P<0.001]. Conclusions:CRAMP protein can inhibit the damage of cardiac microvascular endothelial cells by increasing eNOS-mediated NO signal pathway.

Effect of Xueshuan Xinmaining tablet combined with metoprolol on cardiac enzymes and troponin in patients with coronary heart disease after percutaneous coronary intervention / 中国基层医药

Objective:To investigate the effects of Xueshuan Xinmaining tablet combined with metoprolol on creatine kinase-MB and troponin in patients with coronary heart disease after percutaneous coronary intervention. Methods:A total of 104 patients with coronary heart disease who received percutaneous coronary intervention in Zhoushan Hospital from March 2020 to March 2021 were included in this study. They were randomly divided into observation and control groups ( n = 52/group). The control group was give metoprolol (oral, 25 mg once,3 times/day). The observation group was given Xueshuan Xinmaining tablet (2 tablets once, 3 times per day) based on medication given in the control group. Two groups were treated for 1 month. Clinical efficacy, changes in vascular endothelial function and serum inflammatory factors post-treatment relative to those before treatment, and the incidence of adverse reactions were compared between the two groups. Results:Total response rate in the observation group was significantly higher than that in the control group [86.54% (45/52) vs. 67.31% (35/52), χ2 = 4.99, P < 0.05]. After treatment, nitric oxide in the observation group was significantly higher than that in the control group [(67.23 ± 9.52) μmol/L vs. (60.49 ± 9.71) μmol/L, t = 3.57, P < 0.001]. Endothelin in the observation group was significantly lower than that in the control group [(53.12 ± 7.28) ng/L vs. (61.25 ± 8.36) ng/L, t = 5.28, P < 0.001]. Tumor necrosis factor α, C-reactive protein and interleukin-6 in the observation group were (39.51 ± 6.37) μg/L, (4.13 ± 1.02) mg/L, and (19.43 ± 2.57) μg/L, respectively, which were significantly lower than (51.37 ± 7.28) μg/L, (5.62 ± 1.15) mg/L, (26.16 ± 3.19) μg/L in the control group ( t = 8.84, 6.99, 11.84, all P < 0.05). Creatine kinase-MB and troponin in the observation group were (30.18 ± 5.89) U/L and (7.32 ± 1.12) ng/L, respectively, which were significantly lower than (41.74 ± 6.76) U/L and (9.63 ± 1.45) ng/L in the control group, respectively ( t = 9.29, 9.09, both P < 0.05). No serious adverse reactions occurred during the treatment period in each group. Conclusion:Xueshuan Xinmaining tablet combined with metoprolol exhibit remarkable therapeutic effects on patients with coronary heart disease subjected to percutaneous coronary intervention. The combined therapy can greatly reduce inflammatory reaction and decrease creatine kinase-MB level and improve vascular endothelial function.

Bone morphogenetic protein 4 promotes the proliferation and migration of retinal vascular endothelial cells / 中华眼底病杂志

Objective:To observe the effect of bone morphogenetic protein 4 (BMP4) on the proliferation and migration of human retinal microvascular endothelial cells (hRMEC) under oxidative stress.Methods:The hRMEC cultured in vitro were divided into control group, 4-hydroxynonenal (HNE) treatment group (4-HNE group), 4-HNE+BMP4 group (BMP4 group). Cell culture medium of 4-HNE treatment group was added with 10 μmmol/L 4-HNE; cell culture of BMP4 group was cultured with 10 μmmol/L 4-HNE, and after stimulation for 6 h, 100 ng/ml recombinant human BMP4 was added. The effects of 4-HNE and BMP4 on hRMEC viability was detected by thiazole blue colorimetric method. The effects of 4-HNE and BMP4 on cell migration was determined by cell scratch test. The relative expression of BMP4 mRNA in the cells of the control group and 4-HNE treatment group and the mRNA expression of the control group, the fibronectin (FN) of BMP4 group, laminin (Laminin), α-smooth muscle contractile protein (α-SMA), and collagen type Ⅰ (Collagen Ⅰ), vascular endothelial growth factor (VEGF), and connective tissue growth factor (CTGF) were detected by real-time quantitative polymerase chain reaction (qRT-PCR). Western blot was used to detect the relative expression of BMP4 protein in the control group and 4-HNE group. The control group and 4-HNE group were compared by t test. Results:Compared with the control group, cell viability ( t=12.73, 16.26, P=0.000 2, <0.000 1), cell migration rate ( t=28.17, 37.48, P<0.000 1, <0.000 1) in 4-HNE group and BMP4 group were significantly increased, and the difference was statistically significant; the relative expression of BMP4 mRNA and protein in the 4-HNE group was significantly increased, and the difference was statistically significant ( t=16.36, 69.35, P=0.000 1, <0.000 1). The qRT-PCR test results showed that compared with the control group, the relative expression of VEGF, FN, Laminin, α-SMA, Collagen Ⅰ, and CTGF mRNA in the cells of the BMP4 group was significantly increased, and the difference was statistically significant ( t=10.61, 17.00, 14.85, 7.78, 12.02, 10.61, P=0.0004, <0.000 1, 0.000 1, 0.001 5, 0.000 1, 0.000 4). Conclusion:BMP4 can induce the proliferation and migration of hRMEC; it can also regulate the expression of angiogenesis factors and fibrosis-related factors in hRMEC.

miR-191 inhibits retinal vascular endothelial cell proliferation and angiogenesis / 中华眼底病杂志

Objective:To observe the inhibitory effect of lentivirus (LV)-mediated miR-191 on the proliferation and angiogenesis of human retinal vascular endothelial cells (hREC) cultured in vitro.Methods:The hREC cell lines were cultured in vitro and divided into control group, hypoxia group, LV-empty vector (LV-vector) group, and LV-miR-191 (LV-191) group. The LV-vector group and LV-191 group were transferred to the corresponding lentiviral vector respectively. Flow cytometry was used to detect cell transfection efficiency. Cell Counting Kit-8 (CCK-8) test was used to detect cell proliferation ability. Scarification test and invasion chamber (Transwell) test were used to detect cell migration ability. Matrigel test was used to detect cell lumen formation ability. Real-time quantitative polymerase chain reaction (qPCR) was used to detect the relative expression of miR-191 and relative mRNA expression of its downstream target genes p21, vascular endothelial growth factor (VEGF), cell division protein kinase (CDK) 6, cyclin-D1 (Cyclin D1). Independent sample t test was used for pairwise comparison. Results:The results of flow cytometry showed that the transfection efficiency of cells in the control group and the LV-191 group were 0.615% and 99.400%, respectively. The results of CCK-8, scarification, Transwell and Matrigel test showed that, compared with the control group, the number of cell proliferation ( t=6.130, 4.606), the cell mobility ( t=4.910, 6.702), the number of stained cells on the microporous membrane ( t=7.244, 6.724) and the lumen formation ability cells ( t=8.345, 9.859) were significantly increased in the hypoxia group and the LV-vector group ( P<0.01), while the LV-191 group showed completely opposite performance ( t=14.710, 6.245, 5.333, 5.892; P≤0.01). The qPCR test results showed that, compared with the control group and the LV-vector group, the relative expression of miR-191 mRNA in the cells of the LV-191 group was significantly up-regulated ( t=44.110, 42.680), the relative expression of Cyclin D1 mRNA ( t=29.940, 14.010) and CDK6 mRNA ( t=15.200, 7.645) decreased significantly, and the difference were statistically significant ( P<0.01); the relative expression of p21 mRNA increased, however, the difference was not statistically significant ( t=2.013, 2.755; P>0.05). There was no significant difference in the relative expression of VEGF mRNA in the 4 groups of cells ( F=0.966, P>0.05). Conclusions:LV-191 can inhibit the proliferation, migration and tubing of hREC by up-regulating p21 and down-regulating CDK6 and Cyclin D1.

Associação entre Infecção por Heicobacter Pylori e Hipertensão Arterial Sistêmica: Metanálise / Association between Helicobacter Pylori Infection and Systemic Arterial Hypertension: A Meta-Analysis

Resumo Fundamento: Estudos epidemiológicos recentes demonstraram que alterações na microbiota e seus metabólitos estão associadas à hipertensão arterial sistêmica. A Helicobacter pylori (H. pylori) é um dos patógenos bacterianos mais comuns, e a possível associação entre a infecção por H. pylori e a hipertensão é controversa. Objetivos: Este estudo teve o objetivo de esclarecer a associação entre eles e proporcionar uma nova base teórica para detectar a patogênese da hipertensão. Métodos: Foram selecionados estudos caso-controle e transversais sobre a associação entre H. pylori e hipertensão, publicados de 1996 a 2019 indexados nos bancos de dados PubMed, Google Scholar, Chinese Wan Fang Data, e Chinese National Knowledge Infrastructure (CNKI). As razões de chance (RC) combinadas e o intervalo de confiança (IC) 95% foram estimados. O I² foi realizado para avaliar a heterogeneidade estatística. O viés de publicação foi avaliado utilizando-se os testes de Beggs e de Egger. Os dados extraídos foram analisados no software Stata 12.0. A significância estatística foi definida com um p-valor < 0,05. Resultados: Foram cadastrados 17 estudos envolvendo 6376 casos de hipertensão e 10850 controles. A taxa de infecção por H. pylori em pacientes hipertensos e em controles foi de 64,9% e 56,3%, respectivamente. Foi demonstrada uma associação significativamente positiva entre a infecção por H. pylori e a hipertensão, com uma RC global de 2,07 (IC 95%: 1,46-2,94; p < 0,05). A análise de subgrupos revelou que a prevalência de infecção por H. pylori foi associada à hipertensão na região da Ásia e no grupo de caso-controle, as RC (IC 95%) foram 2,26 (1,51-3,38) e 2,53 (1,72-3,72), respectivamente. Depois de estratificar por métodos de detecção, ainda existiam diferenças entre os subgrupos (todos p < 0,05). Conclusão: Esta metanálise indicou que a infecção por H. pylori está associada positivamente à hipertensão.

Abstract Background: Recent epidemiological studies have shown that alterations in microbiota and its metabolites are associated with systemic arterial hypertension. Helicobacter pylori (H. pylori) is one of the most common bacterial pathogens, and the potential association between H. pylori infection and hypertension are controversial. Objective: This study aimed to clarify their association and provide a new theoretical basis for uncovering the pathogenesis of hypertension. Methods: Case-control and cross-sectional studies on the association between H. pylori and hypertension published from 1996 to 2019 indexed in PubMed, Google Scholar, Chinese Wan Fang Data, and Chinese National Knowledge Infrastructure (CNKI). The pooled odds ratios (OR) and 95% confidence interval (CI) were estimated. I2 was performed to evaluate the statistical heterogeneity. Publication bias was evaluated using Begg's and Egger's test. The extracted data was analyzed in Stata 12.0. Statistical significance was defined as p-value < 0.05. Results: A total of 17 studies involving 6,376 cases of hypertension and 10,850 controls were enrolled. H. pylori infection rate in hypertension patients and controls were 64.9% and 56.3%, respectively. A significantly positive association was shown between H. pylori infection and hypertension with an overall OR of 2.07 (95% CI: 1.46-2.94; p < 0.05). Subgroup analysis revealed that the prevalence of H. pylori infection was associated with hypertension in the region of Asia and the case-control group, ORs (95% CI) were 2.26 (1.51-3.38) and 2.53 (1.72-3.72), respectively. After stratifying by detection methods, differences still existed in subgroups (all p < 0.05). Conclusion: This meta-analysis indicated that H. pylori infection is positively associated with hypertension.

O efeito da atorvastatina + aspirina na função endotelial difere com a idade em pacientes com HIV: Um estudo de caso-controle / The effect of atorvastatin + aspirin on the endothelial function differs with age in patients with HIV: A case-control study

Resumo Fundamento Pacientes com HIV têm maior probabilidade de apresentar doenças cardiovasculares quando comparados à população em geral. Objetivo Este foi um estudo de caso-controle que teve como objetivo avaliar quais fatores estavam associados a uma redução na espessura médio-intimal da carótida (IMT) da carótida e ao aumento na dilatação mediada por fluxo (DMF) da artéria braquial em pacientes com HIV que receberam atorvastatina + aspirina por um período de 6 meses. Métodos Foi realizada uma análise secundária de um ensaio clínico, que incluiu pessoas vivendo com HIV e baixo risco cardiovascular. Um total de 38 pacientes alocados para o braço de intervenção e tratados por 6 meses com uma combinação de atorvastatina + aspirina foram incluídos. Todos os participantes foram submetidos a ultrassonografia da carótida e da artéria braquial, tanto no início quanto no final do estudo. Os casos que responderam com aumento >10% da dilatação braquial (DMF) e redução da espessura médio-intimal da carótida (IMT) foram considerados casos, e aqueles que não responderam foram considerados controles. Avaliamos os fatores associados às respostas positivas obtidas através da IMT e DMF. Resultados A redução do IMT não se associou significativamente a nenhum dos fatores de risco avaliados: idade (p = 0,211), sexo (p = 0,260), tabagismo (p = 0,131) ou tempo de diagnóstico do HIV (p = 0,836). Um aumento na DMF foi significativamente associado com a idade entre aqueles na faixa etária de 40-59 anos, p = 0,015 (OR = 4,37; IC 95%: 1,07-17,79). Conclusões Os indivíduos mais velhos foram mais propensos a apresentar um aumento na DMF após 6 meses de tratamento com atorvastatina + aspirina.

Abstract Background Patients with HIV are more likely to present with cardiovascular disease when compared to the general population. Objective This was a case-control study that aimed to assess which factors were associated with a reduction in the carotid intima-media thickness (IMT) and an increase in the brachial artery flow-mediated dilation (FMD) in HIV patients who received atorvastatin + aspirin during a period of 6 months. Methods A secondary analysis of a clinical trial was conducted, which included people living with HIV infection and low cardiovascular risk. A total of 38 patients allocated to the intervention arm and treated for 6 months with a combination of atorvastatin + aspirin were included. All participants underwent a carotid and brachial artery ultrasound, both at the beginning and the end of the study. Cases that responded with an increase of >10% of the brachial dilatation (FMD) and reduction of the carotid intima-media thickness (IMT) were considered cases, and those who did not respond were considered controls. We assessed the factors associated with the positive responses obtained through IMT and FMD. Results A reduction in the IMT was not significantly associated with any of the evaluated risk factors: age (p=0.211), gender (p=0.260), smoking (p=0.131) or time since HIV diagnosis (p=0.836). An increase in the FMD was significantly associated with age amongst those in the 40-59 age group, p = 0.015 (OR = 4.37; 95% CI: 1.07-17.79). Conclusions Older individuals were more likely to present with an increased FMD after 6 months of treatment with atorvastatin + aspirin.

Pacientes Naïve Infectados por HIV Apresentam Disfunção Concomitante com Diminuição de Anticorpos Naturais contra Autoantígenos Derivados da Apolipoproteína B Definidos / HIV-Infected Naïve Patients Exhibit Endothelial Dysfunction Concomitant with Decreased Natural Antibodies Against Defined Apolipoprotein B Autoantigens

Fundamento: Fatores de risco definidos para HIV e tradicionais podem estar associados a um aumento de eventos cardiovasculares. Estudos recentes sugerem que a resposta imune humoral à LDL modificada pode estar associada ao processo de aterosclerose. Objetivos: Avaliar a presença de anti-LDL oxidada e de peptídeos derivados da Apolipoproteína B no sangue, bem como sua associação à função endotelial na infecção por HIV. Métodos: Este estudo incluiu consecutivamente sujeitos com idade, sexo e dados demográficos correspondentes em dois grupos: (1) indivíduos infectados com HIV e naïve para terapia antiviral e (2) indivíduos não infectados. A aterosclerose subclínica foi avaliada pela espessura íntima-média, utilizando-se a ultrassonografia das artérias carótidas. A função endotelial foi determinada pela dilatação mediada por fluxo (DMF) da artéria braquial por ultrassonografia. Os níveis de autoanticorpos (IgM, IgG) de lipoproteínas de baixa densidade antioxidadas (LDL-ox), fragmentos de peptídeos antiapolipoproteína B (peptídeos ApoB-D e 0033G-Cys), e citocina foram avaliados por meio de ELISA. Resultados: Os resultados deste estudo não mostraram diferenças na aterosclerose subclínica entre os grupos. Entretanto, os sujeitos infectados com HIV apresentaram uma DMF mais baixa, em comparação com os sujeitos não infectados. Portanto, os sujeitos infectados com HIV apresentaram níveis mais altos de citocinas inflamatórias, títulos de IgG anti-LDL-ox, e IgG anti-ApoB-D. Em contraste, títulos de IgM anti-ApoB-D foram mais baixos em indivíduos infectados com HIV e associados a funções endoteliais diminuídas. Conclusões: Os resultados deste estudo mostram que a infecção por HIV, em sujeitos naïve, está associada à disfunção endotelial e à diminuição de anticorpos naturais para antígenos Apo-B.

Abstract Backgorund: Traditional and HIV-defined risk factors may be associated with an increase in cardiovascular events. Recent studies have suggested that the humoral immune response to modified LDL may be associated with the process of atherosclerosis. Objectives: To evaluate the presence of anti-oxLDL and apolipoprotein B-derived peptides in the blood, and their association with the endothelial function in HIV-infection. Methods: This study consecutively included subjects matched for age, gender, and demographic data in two groups: (1) HIV-infected and naïve for antiviral therapy and (2) uninfected individuals. Subclinical atherosclerosis was assessed by intimal-media thickness, using ultrasonography of the carotid arteries. Endothelial function was determined by flow-mediated dilatation (FMD) of the brachial artery by ultrasonography. Autoantibodies (IgM, IgG) anti-oxidized low-density lipoprotein (oxLDL), anti-apolipoprotein B-peptide fragments (ApoB-D and 0033G-Cys peptides), and cytokine levels were evaluated by ELISA. Results: This study's results showed no difference in subclinical atherosclerosis between groups; however, HIV-infected subjects showed a lower FMD, when compared to non-infected subjects. Therefore, HIV-infected subjects showed higher levels of inflammatory cytokines, titers of IgG anti-oxLDL, and IgG anti-ApoB-D. In contrast, titers of IgM anti-ApoB-D were lower in HIV-infected individuals and associated with reduced endothelial functions. Conclusions: This study's results show that HIV infection, in naïve subjects, is associated with endothelial dysfunction and a decline of natural antibodies to apo-B antigens.