Adjuvant-induced arthritis promotes vascular hyporesponsiveness to phenylephrine through a nitric oxide-related mechanism

Arthritis has important cardiovascular repercussions. Phenylephrine-induced vasoconstriction is impaired in rat aortas in the early phase of the adjuvant-induced arthritis (AIA), around the 15th day post-induction. Therefore, the present study aimed to verify the effects of AIA on hyporesponsiveness to phenylephrine in rat aortas. AIA was induced by intradermal injection of Mycobacterium tuberculosis (3.8 mg/dL) in the right hind paw of male Wistar rats (n=27). Functional experiments in isolated aortas were carried out 15 days after AIA induction. Morphometric and stereological analyses of the aortas were also performed 36 days after the induction of AIA. AIA did not promote structural modifications in the aortas at any of the time points studied. AIA reduced phenylephrine-induced contraction in endothelium-intact aortas, but not in endothelium-denuded aortas. However, AIA did not change KCl-induced contraction in either endothelium-intact or denuded aortas. L-NAME (non-selective NOS inhibitor), 1400W (selective iNOS inhibitor), and ODQ (guanylyl cyclase inhibitor) reversed AIA-induced hyporesponsiveness to phenylephrine in intact aortas. 7-NI (selective nNOS inhibitor) increased the contraction induced by phenylephrine in aortas from AIA rats. In summary, the hyporesponsiveness to phenylephrine induced by AIA was endothelium-dependent and mediated by iNOS-derived NO through activation of the NO-guanylyl cyclase pathway.

Termografía infrarroja en la detección de la función endotelial normal / Infrared thermography for normal endothelial function screening

Resumen Antecedentes: La sospecha de disfunción endotelial (DE) permitirá prevenir la aterosclerosis acelerada y la muerte prematura. Objetivo: Establecer la utilidad de la termografía en el cribado de la función endotelial en adultos con factores de riesgo cardiovascular. Material y métodos: Estudio transversal analítico de prueba diagnóstica. El incremento del diámetro de la arteria braquial < 11 % a un minuto posisquemia significó probable DE, confirmada si el diámetro fue ≥ 11 % posnitroglicerina sublingual. Se obtuvieron fotografías termográficas al minuto de la región palmar. Se aplicó estadística descriptiva, curva ROC, pruebas U de Mann-Whitney, chi cuadrada o exacta de Fisher. Resultados: Se incluyeron 38 sujetos, mediana de edad de 50 años, con 624 mediciones termográficas; nueve presentaron DE (vasodilatación mediada por flujo de 2.5 %). El mejor punto de corte para la función endotelial normal en sujetos con factores de riesgo cardiovascular fue ≥ 36 °C al minuto de isquemia, con sensibilidad de 85%, especificidad de 70%, valores predictivos positivo y negativo de 78 y 77%, área bajo la curva de 0.796, razón de verisimilitud positiva de 2.82 y razón de verisimilitud negativa de 0.22. Conclusión: La medición de la temperatura en la región palmar mediante termografía infrarroja ≥ 36 °C tras un minuto de isquemia es práctica, no invasiva y económica para el cribado de la función endotelial normal en adultos con factores de riesgo cardiovascular.

Abstract Background: Endothelial dysfunction (ED) suspicion will allow to prevent accelerated atherosclerosis and premature death. Objective: To establish the usefulness of thermography for endothelial function screening in adults with cardiovascular risk factors. Material and methods: Cross-sectional, analytical diagnostic test. A brachial arterial diameter (BAD) increase <11 % at one-minute post-ischemia meant probable ED and was confirmed if BAD was ≥ 11 % post-sublingual nitroglycerin. Thermographic photographs of the palmar region were obtained at one minute. Descriptive statistics, ROC curve, Mann-Whitney’s U-test, chi-square test, or Fisher’s exact test were used. Results: Thirty-eight subjects with a median age of 50 years, and with 624 thermographic measurements were included. Nine had ED (flow-mediated vasodilation (FMV): 2.5 %. The best cutoff point for normal endothelial function in subjects with cardiovascular risk factors was ≥ 36 °C at one minute of ischemia, with 85 % sensitivity, 70 % specificity, positive and negative predictive values of 78 and 77 %, area under the curve of 0.796, LR+ 2.82, LR- 0.22. Conclusions: An infrared thermography-measured temperature in the palmar region greater than or equal to 36 °C after one minute of ischemia is practical, non-invasive, and inexpensive for normal endothelial function screening in adults with cardiovascular risk factors.

24-dehydrocholesterol reductase ameliorates senescence-related dysfunction of vascular endothelial cells / 中华老年医学杂志

Objective:To investigate the role of 24-dehydrocholesterol reductase(DHCR24)in doxorubicin-induced senescence-related dysfunction of vascular endothelial cells.Methods:Human umbilical vein endothelial cells(HUVECs)were induced with 0.05 μM doxorubicin for 48 h to establish a stress-triggered premature senescence model.The lentiviral transfection method was employed to achieve DHCR24 overexpression in HUVECs.Cell senescence was evaluated by β-galactosidase staining and Western blot to detect the expression of the senescence-related molecules cyclin-dependent kinase inhibitor 1A(P21)and nicotinamide adenine dinucleotide dependent histone deacetylase 1(SIRT1).Western blot was performed to detect DHCR24 and endothelial nitric oxide synthase(eNOS)expression during endothelial senescence.DAF-FM DA(an NO fluorescent probe)was used to detect intracellular NO production.Results:In the stress-triggered premature senescence model of HUVECs induced by doxorubicin, the expression of the senescence marker P21 was up-regulated( t=19.44, P<0.01), SIRT1 was down-regulated( t=10.10, P<0.01, and the expression of DHCR24 was down-regulated( t=5.946, P<0.01), compared with the control group.Meanwhile, eNOS and NO expression was inhibited( t=11.26, P<0.01; t=10.83, P<0.01).After DHCR24 overexpression, compared with the control stimulation group, the overexpression stimulation group showed that DHCR24( F=72.10, P<0.01)was up-regulated.DHCR24 overexpression alleviated the doxorubicin-induced decrease in eNOS and NO( F=5.797, P<0.05; F=45.12, P<0.01), compared with the control group. Conclusions:DHCR24 may mitigate doxorubicin-induced senescence-related vascular endothelial dysfunction by modulating the eNOS/NO signaling pathway.

The effects of aerobic exercise on endothelial nitric oxide synthase uncoupling in the myocardium of spontaneously hypertensive rats / 中华物理医学与康复杂志

Objective:To observe any effect of regular aerobic exercise on cardiac remodeling in spontaneously hypertensive rats (SHR) and explore the mechanism of endothelial nitric oxide synthase (eNOS) uncoupling.Methods:Thirty 6-week-old healthy male SHR were divided into a sedentary group and an exercise group, each of 15. Another ten age- and sex-matched Wistar-Kyoto rats were set as a normal control group. The animals in the normal control and sedentary groups were fed quietly in their cages, and those in the exercise group performed moderate intensity treadmill exercise for 8 weeks (5 times per week). Forty-eight hours after the last training, echocardiography was applied to document cardiac structure and function in both groups. Wheat germ agglutinin staining and Picrosirius Red staining were used to obtain the cardiomyocyte cross sectional areas (CSAs) and interstitial collagen volume fractions (CVFs) of all of the mice. The rates of cardiomyocyte apoptosis were measured using TUNEL staining, and myocardial tetrahydrobiopterin (BH4) content was measured using high-performance liquid chromatography. Total eNOS, eNOS dimer and eNOS monomer protein expression in the myocardia were detected using western blotting.Results:Compared with the normal control group, the left ventricular wall thickness (LVWT), myocardial CSA, CVF, apoptosis of cardiomyocytes and eNOS monomer levels were significantly higher in the sedentary group, on average. But the end-diastolic diameter (LVEDD) and ejection fraction (LVEF) of the left ventricle and the levels of eNOS dimer and myocardial BH4 and the eNOS dimer/monomer ratio tended to be lower. Comparing the exercise group with the sedentary group, the average LVEDD, LVEF, eNOS dimer, eNOS dimer/monomer ratio and myocardial BH4 content were significantly higher in the exercise group, but the myocardial CVF, cardiomyocyte apoptosis and eNOS monomer levels were significantly lower. LVWT and CSA were not significantly different. There were no significant differences in the total eNOS protein levels among the three groups.Conclusion:Regular aerobic exercise might improve cardiac remodeling in cases of spontaneous hypertension regulating eNOS uncoupling, at least in rats.

Network Pharmacological Analysis and Experimental Verification of the Mechanism of Gastrodiae Rhizoma-Salviae Miltiorrhizae Radix et Rhizoma Drug Pair in the Treatment of Hypertension / 中药新药与临床药理

Objective To investigate the mechanism of Gastrodiae Rhizoma-Salviae Miltiorrhizae Radix et Rhizoma drug pair in the treatment of hypertension based on the network pharmacology method and animal experiment verification.Methods(1)TCMSP,BATMAN and TCMIP databases were used to screen the active components and targets of Gastrodiae Rhizoma-Salviae Miltiorrhizae Radix et Rhizoma drug pair.The hypertension-related targets were obtained by searching the Drugbank,Genecard,TTD and Disgenet databases.The intersection(common target)of the active component target and the target related to hypertension disease was taken,and the obtained intersection target was the potential target of Gastrodiae Rhizoma-Salviae Miltiorrhizae Radix et Rhizoma drug pair for the treatment of hypertension.The active ingredients and their targets of Gastrodiae Rhizoma-Salviae Miltiorrhizae Radix et Rhizoma drug pair were imported into Cytoscape 3.9.1 software to construct a'Chinese medicines-active ingredients-targets'network and screen key active ingredients.The protein-protein interaction(PPI)network of potential targets was constructed to screen potential core targets.The Metascape platform was used to analyze the GO function and KEGG pathway enrichment of potential targets.The key active components and potential core targets were selected for molecular docking verification.(2)Thirty male spontaneously hypertensive rats(SHR)were randomly divided into model group,western medicine group(Candesartan Cilexetil,0.72 mg·kg-1)and low-,medium-and high-dose groups of Gastrodiae Rhizoma-Salviae Miltiorrhizae Radix et Rhizoma(2.25,4.50,9.00 g·kg-1).Another male WKY rats were selected as blank group,with 6 rats in each group,once a day for 8 weeks.The systolic blood pressure of rat tail artery was detected before administration and 2,4,6 and 8 weeks after drug intervention.The pathological changes of thoracic aorta were observed by HE staining.The protein expression levels of GRP78,CHOP and Caspase-12 in aorta abdominalis were detected by Western Blot.Results(1)A total of 83 active components of Gastrodiae Rhizoma-Salviae Miltiorrhizae Radix et Rhizoma were obtained,and 158 potential targets(intersection targets)for the treatment of hypertension were screened out.Five key active ingredients:p-hydroxybenzoic acid,4-hydroxybenzylamine,tanshinone I,tanshinone,γ-sitosterol;6 potential core targets:IL6,TNF,CASP3,JUN,PTGS2,IL1B;GO functional enrichment analysis obtained 1 826 biological process items,89 cell component items,and 199 molecular function items.KEGG pathway enrichment analysis obtained 186 pathways,mainly involving neuroactive ligand-receptor interaction,calcium signaling pathway,inflammatory response(such as TNF and MAPK signaling pathway),vascular protection(such as HIF-1 and cAMP signaling pathway),oxidative stress(such as PI3K-Akt signaling pathway)and other signaling pathways.Tanshinone I and tanshinone had strong binding force to 6 potential core targets,and γ-sitosterol had strong binding force to IL6,CASP3,JUN,PTGS2 and IL1B.(2)Compared with the blank group,the systolic blood pressure of the model group was significantly increased(P＜0.01).The thoracic aortic endothelial injury was obvious,the endothelial cell morphology was abnormal,swelling and exfoliated cells could be seen,the intima of the tissue was disordered,the intima structure was incomplete,and the intima was thickened.The protein expressions of GRP78,CHOP and Caspase-12 in abdominal aorta were significantly increased(P＜0.01).Compared with the model group,the systolic blood pressure of the rats in the administration group was significantly decreased(P＜0.01);the injury of thoracic aorta was alleviated,and the morphology,intima structure and thickness of endothelial cells were improved to varying degrees.The protein expressions of GRP78,CHOP and Caspase-12 in abdominal aorta were significantly decreased(P＜0.01).Conclusion Gastrodiae Rhizoma-Salviae Miltiorrhizae Radix et Rhizoma drug pair may act on core targets such as IL6,TNF,CASP3,JUN,PTGS2,and IL1B through key active components such as p-hydroxybenzoic acid,tanshinone,and γ-sitosterol,and regulate key signaling pathways such as TNF signaling pathway,MAPK signaling pathway,PI3K-Akt signaling pathway,and PERK signaling pathway to improve vascular endothelial dysfunction,inhibit endoplasmic reticulum stress,and lower blood pressure.

In Vitro and in Vivo Evaluation of Scutellarin-phospholipid Complex Nanoemulsion and Analysis of Its Activity in Ameliorating LPS-induced Vascular Endothelial Injury / 中国实验方剂学杂志

ObjectiveTo evaluate some properties of scutellarin-phospholipid complex nanoemulsion（SCU-PC-NE）， such as release， cell uptake and tissue distribution， and to investigate its effect on ameliorating lipopolysaccharide（LPS）-induced vascular endothelial injury. MethodSCU-PC-NE was prepared by weighting SCU-PC， ethyl oleate， Kolliphor HS15， 1，2-propylene glycol（50， 400， 514.3， 85.7 mg）， respectively. And the appearance of SCU-PC-NE was observed by transmission electron microscope， the average paticle size and Zeta potential were measured by nanopotential particle size analyzer. The cumulative release of SCU-PC-NE in vitro was measured by dynamic dialysis， thiazolyl blue（MTT） colorimetric assay was used to investigate the effect of SCU-PC-NE on the viability of human umbilical vein endothelial cells（HUVECs）， the inverted fluorescence microscope and flow cytometry were used to investigate cell uptake of HUVECs by SCU-PC-NE in vitro using coumarin 6 as a fluorescent probe， the tissue distribution of DiR/SCU-PC-NE labeled by near infrared fluorescent dyes was obeserved by small animal in vivo imaging system. The inflammation injury model was established by co-incubation with LPS（1 mg·L-1） and HUVECs， the effect of SCU-PC-NE on the levels of interleukin（IL）-1β and IL-6 were determined by enzyme-linked immunosorbent assay（ELISA）， 18 Kunming male mice were randomly divided into blank group， model group， blank preparation group（equivalent to high dose group）， SCU group and SCU-PC-NE low and high dose groups（5， 10 mg·kg-1）， 3 mice in each group， and the drug administration groups were administered once in the tail vein at the corresponding dose every 48 h， equal volume of normal saline was given to the blank group and the model group， and the drug was administered for 4 consecutive times. Except for the blank group， the endothelial inflammatory injury was induced by intraperitoneal injection of LPS（10 mg·kg-1） at 12 h before the last administration in each group. Hematoxylin-eosin（HE） staining was used to investigate the effect of SCU-PC-NE on the histopathological changes in the thoracic aorta of mice. ResultThe appearance of SCU-PC-NE displayed pale yellow milky light， mostly spherical with rounded appearance and relatively uniform particle size distribution， with the average particle size of 35.31 nm， Zeta potential of 7.23 mV， and the encapsulation efficiency of 75.24%. The cumulative release in vitro showed that SCU-PC-NE exhibited sustained release properties compared with SCU. The cell viability of SCU-PC-NE was >90% at a concentration range of 1.05-8.4 mg·L-1. The results of cellular uptake experiments showed that the cellular uptake ability of SCU-PC-NE was significantly enhanced when compared with the SCU group（P<0.01）. Compared with normal mice， the results of tissue distribution showed that the fluorescence intensity of DiR/SCU-PC-NE was significantly enhanced in the spleen， kidney， brain and thoracic aorta of mice at different time points after intraperitoneal injection of LPS（P<0.05， P<0.01）， especially in thoracic aorta. ELISA results showed that the levels of IL-1β and IL-6 in the model group were significantly increased when compared with the blank group（P<0.05， P<0.01）， and compare with the model group， all administration groups significantly down-regulated IL-1β level， with the strongest effect in the SCU-PC-NE high-dose group（P<0.01）， and all administration groups significantly down-regulated IL-6 level， with the strongest effect in the SCU-PC-NE low-dose group（P<0.05）. Compare with the blank group， the results of HE staining showed that the endothelial cells were damaged， the elastic fibers were broken and arranged loosely in the model group， although similar vascular injury could be observed in the blank preparation group， SCU group and SCU-PC-NE low-dose group， the vascular endothelial damage was significantly reduced in the high-dose group of SCU-PC-NE， which had a better effect than that in the SCU group. ConclusionSCU-PC-NE can promote the uptake of drugs by endothelial cells and effectively enriched in the site of vascular endothelial injury caused by LPS， suggesting that it has a protective effect on vascular endothelial injury and is a good carrier of SCU.

Hot topics in endothelial keratoplasty research and future research directions / 中华实验眼科杂志

Objective:To investigate the distribution of literature and hot topics of endothelial keratoplasty (EK) in recent years.Methods:Using the Science Citation Index Expanded and Social Science Citation Index in Web of Science Core Collection database as the data sources, the literature on EK from 2011 to 2021 was collected and the annual volume of published original research articles on EK, the top 10 authors and journals with the highest number of published articles, and the top 10 cited articles were analyzed.The author and keyword data of the collected relevant literature were imported into VOSviewer software for visual analysis.Results:A total of 1 454 articles were included.The cumulative volume of articles increased linearly from 2011 to 2021, the United States was the country that published the most EK-related literature, with Melles GRJ ranking first in terms of the highest number of published articles and the highest number of citations, and Cornea ranking first in terms of the total number of publications.Hot topics of EK mainly focused on the comparison of postoperative effects of different procedures and the management of complications.Keywords were grouped into 5 clusters, the evaluation of the postoperative visual quality of EK, the high-risk factors for postoperative complications and transplantation failure, the indications and surgical techniques of Descemet membrane endothelial keratoplasty (DMEK), corneal graft issues including graft preparation, endothelial cell loss and management of graft dislocation, and the changing trend of corneal transplantation.The most frequently occurring keyword was DMEK, with a total of 559 occurrences, indicating that DMEK was the focus of EK-related research.Since 2017, the frequency of keywords such as DMEK, previous trabeculectomy, graft preparation, stability, safety, and air has increased, suggesting that the treatment of patients with a history of glaucoma surgery, the safety and stability of surgery, and the preparation of big-bubble graft had been the focus of research in recent years. Conclusions:Graft preparation, the management of complicated cases, the treatment of glaucoma in patients with corneal endothelial diseases and the evaluation of the long-term postoperative effects of big-bubble graft preparation may become the future EK research direction.

Advances in the tissue-engineered corneal endothelial transplantation / 中华实验眼科杂志

Due to the serious shortage of corneal donor, the development of penetrating keratoplasty and corneal endothelial transplantation is severely restricted in clinical practice.The root cause is the limited proliferation capacity of healthy corneal endothelial cells.With the continuous development of tissue engineering technology and cell engineering technology, the research of tissue-engineered cornea has made some progress. In vitro culture of corneal endothelial cells with high density and healthy endothelial function for transplantation is a hot topic in current tissue engineering research.The keys of tissue-engineered corneal endothelial technology include seed cells, vector materials and the strategy of cell transplantation.At present, many research teams domestic and abroad have reported that the source of seed cells includes human corneal endothelial cells, stem cells, vascular endothelial cells and human amniotic epithelial cells.Vector materials include amniotic membrane, acellular corneal stroma, posterior elastic layer, anterior capsular membrane and various biomaterials.The cultured cells are transplanted by penetrating keratoplasty, corneal endothelial transplantation or anterior chamber injection.This review summarized the latest progress in the research on the source of corneal endothelial seed cells, the selection of vectors and the methods of corneal endothelial transplantation, and summed up the problems faced in the current research and looked forward to its prospects.

Evaluation of the efficacy and safety of phacoemulsification under the IOL protection: a randomized controlled clinical study / 中华实验眼科杂志

Objective:To observe the therapeutic effect of intraocular lens (IOL) protected phacoemulsification (PHACO) in patients with hard nucleus cataract.Methods:A randomized controlled clinical study was conducted.A total of consecutive 120 patients (120 eyes) with hard nucleus cataract of Emery grade Ⅳ or Ⅴ were enrolled from January 2019 to May 2022.The patients were randomly divided into PHACO group receiving routine PHACO, IOL protected PHACO group receiving PHACO under IOL protection, and extracapsular cataract extraction (ECCE) group receiving ECCE, with 40 cases (40 eyes) in each group.Finally, 99 patients completed the follow-up, including 30 cases (30 eyes) in PHACO group, 35 cases (35 eyes) in IOL protected PHACO group, and 34 cases (34 eyes) in ECCE group.The total operation time, intraoperative PHACO time and cumulative energy release of each patient were recorded.The corneal endothelial cell density (ECD), coefficient of variation in endothelial cell area (CV), hexagonal endothelial cell ratio (6A), corneal astigmatism and the number of eyes with different grades of uncorrected visual acuity were measured and compared after 3-month follow-up.The intraoperative and postoperative complications were recorded.This study adhered to the Declaration of Helsinki and was approved by the Ethics Committee of Yanbian University Hospital (NO.2023002).Patients were informed of study content and purpose and signed a consent form before treatment.Results:There was no significant difference in ultrasonic energy and time between PHACO group and IOL protected PHACO group ( P=0.691, 0.982).The total operation time was (38.81±2.73) and (36.45±3.45) minutes in PHACO group and IOL protected PHACO group, significantly shorter than (69.60±4.35) minutes in ECCE group (both at P<0.001).There was no significant difference in age, sex, lens nucleus hardness and other baseline data among the three groups before operation (all at P>0.05).Three months after operation, the number of patients with higher uncorrected visual acuity in PHACO group and IOL protected PHACO group was larger than that in ECCE group ( P=0.006, 0.007).The ECD and 6A in IOL protected PHACO group were (2 155.57±177.88)/mm 2 and (41.31±5.18)%, respectively, which were significantly higher than (1 912.64±224.11)/mm 2 and (36.18±3.27)% in PHACO group, and the CV in IOL protected PHACO group was (50.34±5.90)%, which was lower than (55.67±3.30)% in PHACO group, showing statistically significant differences ( P=0.007, 0.003, 0.005).At 1 week and 3 months after the operation, the corneal astigmatism was significantly lower in IOL-protected PHACO group than in ECCE group, but higher than in PHACO group, and the difference were statistically significant (all at P<0.05). Conclusions:Compared with conventional PHACO, IOL-protected PHACO can effectively reduce the damage of corneal endothelium caused by ultrasonic energy, shorten the operation time and reduce postoperative inflammatory reaction compared with ECCE, and does not significantly increase postoperative corneal astigmatism.IOL-protected PHACO is an effective improved surgical method for patients with hard nucleus cataract.

Biological identity of orbital cavernous venous malformations

ABSTRACT Vascular anomalies comprise a wide spectrum of clinical manifestations related to disturbances in the blood or lymph vessels. They correspond to mainly tumors (especially hemangiomas), characterized by high mitotic activity and proliferation of the vascular endothelium, and malformations, endowed with normal mitotic activity and no hypercellularity or changes in the rate of cell turnover. However, the classifications of these lesions go beyond this dichotomy and consist various systems adapted for and by different clinical subgroups. Thus, the classifications have not reached a consensus and have historically caused confusion regarding the nomenclatures and definitions. Cavernous venous malformations of the orbit, previously called cavernous hemangiomas, are the most common benign vascular orbital lesions in adults. Herein, we have compiled and discussed the various evidences, including clinical, radiological, morphological, and molecular evidence that indicate the non-neoplastic nature of these lesions.

Intravital Microscopy Evidence That Methylene Blue Should Be a Vasopressor-Sparing Agent in Sepsis Vasoplegia

ABSTRACT Microvasculature failure is expected in sepsis and at higher amine concentrations. Therefore, special attention focused individually on microcirculation is needed. Here, we present that methylene blue can prevent leukocytes from adhering to the endothelium in a rat model of lipopolysaccharide-induced endotoxemia. As hypothesis evidence, an intravital microscopy image is presented.

Regulação da Septina 9 na síndrome do desconforto respiratório agudo associado à malária / The role of Septin 9 in regulating vascular permeability in malarial respiratory distress syndrome

A malária é um problema mundial de saúde, com 249 milhões de casos de infecção, ocasionando 608 mil mortes no ano de 2022. Causada pelo gênero Plasmodium, são cinco principais espécies causadoras da malária no ser humano, o Plasmodium malariae, Plasmodium ovale, Plasmodium knowlesi, Plasmodium vivax e Plasmodium falciparum sendo os dois últimos responsáveis pelo maior número de casos clínicos e mortes ao redor do mundo, transmitida pelo mosquito fêmea do gênero Anopheles durante o repasto sanguíneo. Sabe-se ainda que eritrócitos infectados por Plasmodium berghei ANKA causam alteração no citoesqueleto de actina, consequentemente levando a hiperpermeabiliade da barreira endotelial. Em experimentos in vitro, a imunofluorescência, foi observada alteração do citoesqueleto de actina em células estimuladas com eritrócitos parasitados por PbA (EP), em contrapartida, aquelas não estimuladas (NE) e estimuladas com eritrócitos não parasitados por PbA (EnP), não mostraram alterações no mesmo. Nos experimentos in vivo, ao observar dados coletados, sendo estes respiratórios (penh e frequência respiratória) e parasitemia coletados no 7º DPI, foi observado um mesmo padrão entre o experimento 1 e o experimento 2. Os animais infectados com 106 de eritrócitos infectados, foram alocados em dois grupos, sendo eles hiperparasitemia (HP) ou síndrome do desconforto respiratório agudo-associado a malária (SDRA/SDRA-MA) e comparados àqueles não infectados (NI). Os animais NI, não apresentam parasitemia, em contrapartida, os animais SDRA, tem maior parasitemia que os HP, visto que estes têm aumento em sua parasitemia após o 12º DPI, e assim seguem aumentando gradativamente até levar os animais a óbito.O penh tem o mesmo padrão que a parasitemia, os NI com penh mais baixa que os HPs e os SDRA, sendo dentre estes, o grupo SDRA o mais elevado. A frequência respiratória, por sua vez se apresenta mais elevada no grupo NI, sendo o grupo SDRAmenor que o HP, um achado tido como normal, visto que os pulmões de animais com SDRA sofrem maior dano que os HPs. Apesar de não apresentar um valor significativo, as imagens de gel SDS-PAGE (WB) mostram maior concentração da Septina 9 nos animais com SDRA em comparação com os HPs e com os NIs. O mesmo é observado na qRT-PCR, mesmo sem significância estatística, o valor mostrado nos gráficos temmaior concentração nos SDRA. Assim, a Septina 9 está presente nas CEPP, e, mesmo sem significância estatística, da mesma forma que está presente nas amostras de tecido pulmonar utilizadas no WB e qT-PCR. É hipotetizado ainda que esta proteína pode ser ativada e assim sofrer alteração em sua localização intracelular

Malaria is a global health problem, with 249 million cases of infection, causing 608 thousand deaths in the year 2022. Caused by the genus Plasmodium, there are five main species that cause malaria in humans, Plasmodium malariae, Plasmodium ovale, Plasmodium knowlesi, Plasmodium vivax and Plasmodium falciparum, the last two being responsible for the largest number of clinical cases and deaths around the world, transmitted by the female mosquito of the genus Anopheles during blood meal. It is also known that erythrocytes infected by Plasmodium berghei ANKA (PbA) cause changes in the actin cytoskeleton, consequently leading to hyperpermeability of the endothelial barrier. In in vitro experiments, immunofluorescence, changes in the actin cytoskeleton were observed in cells stimulated with erythrocytes parasitized by PbA (EP), in contrast, those not stimulated (NE) and stimulated with erythrocytes not parasitized by PbA (EnP), did not show changes the same. In the in vivo experiments, when observing collected data, these being respiratory (penh and respiratory frequency) and parasitemia collected on the 7th DPI, the same pattern was observed between experiment 1 and experiment 2. Animals infected with 106 infected erythrocytes were allocated into two groups, namely hyperparasitemia (HP) or malaria-associated acute respiratory distress syndrome (ARDS/ARDS-MA) and compared to those not infected (NI). NI animals do not present parasitemia, on the other hand, ARDS animals have greater parasitemia than HP animals, as the latter have an increase in their parasitemia after the 12th DPI, and thus continue to gradually increase until the animals die. the same pattern as parasitemia, NI with lower penh than HPs and ARDS, among these, the ARDS group being the highest. The respiratory rate, in turn, is higher in the NI group, with the ARDS group being lower than the HP, a finding considered normal, given that the lungs of animals with ARDS suffer greater damage than the HPs. Despite not showing a significant value, SDS-PAGE (WB) gel images show a higher concentration of Septin 9 in animals with ARDS compared to HPs and NIs. The same is observed in qRT-PCR, even without statistical significance, the value shown in the graphs has a higher concentration in ARDS. Thus, Septin 9 is present in CEPP, and, even without statistical significance, in the same way that it is present in lung tissue samples used in WB and qT-PCR. It is also hypothesized that this protein can be activated and thus undergo changes in its intracellular location

Biochanin A restored the blood-brain barrier in cerebral ischemia-reperfusion in rats

SUMMARY OBJECTIVE: Blood-brain barrier is a protective layer that regulates the influx and efflux of biological materials for cerebral tissue. The aim of this study was to investigate the effects of Biochanin A on cerebral histopathology and blood-brain barrier immunohistochemically. METHODS: A total of 24 rats were assigned to three groups: sham, ischemia-reperfusion, and ischemia-reperfusion+Biochanin A. Ischemia-reperfusion was performed by occluding the left carotid artery for 2/24 h. Notably, 20 mg/kg Biochanin A was administered to rats for 7 days after ischemia-reperfusion. Blood was collected for malondialdehyde and total oxidant/antioxidant status analysis. Cerebral tissues were processed for histopathology and further for immunohistochemical analysis. RESULTS: Malondialdehyde content with total oxidant status value was significantly increased and total antioxidant status values were significantly decreased in the ischemia-reperfusion group compared with the sham group. Biochanin A treatment significantly improved scores in the ischemia-reperfusion+Biochanin A group. The normal histological appearance was recorded in the cerebral sections of the sham group. Degenerated neurons and vascular structures with disrupted integrity of the cerebral cortex were observed after ischemia-reperfusion. Biochanin A alleviated the histopathology in the cerebrum in the ischemia-reperfusion+Biochanin A group. Ischemia-reperfusion injury decreased the expression of blood-brain barrier in the ischemia-reperfusion group compared to the sham group. Administration of Biochanin A upregulated the blood-brain barrier immunoreactivity in the cerebrum by restoring blood-brain barrier. CONCLUSION: Cerebral ischemia-reperfusion caused an increase in oxidative stress and pathological lesions in the cerebrum. Biochanin A treatment restored the adverse effects of ischemia-reperfusion injury by restoring blood-brain barrier.

Manually prepared lamellae for Descemet stripping endothelial keratoplasty (Pachy-DSEK): comparison of four dissection depths / Lamelas manualmente dissecadas para transplante endotelial de córnea (Pachy-DSEK): comparação entre quatro profundidades de dissecção

ABSTRACT Purpose: This study aimed to compare four depths of manual dissection for the preparation of Descemet stripping endothelial keratoplasty lamellae. Methods: Eye bank corneas were randomized into four groups according to dissection depths: Pachy-100 (incision depth = central corneal thickness-safety margin of 100 µm), Pachy-50 (safety margin of 50 µm), Pachy-0 (no safety margin), and Pachy+50 (incision depth = central corneal thickness + 50 µm). All endothelial lamellae were prepared using a standardized method of manual dissection (Pachy-DSEK). The central, paracentral (3.0-mm zone), and peripheral (6.0-mm zone) lamella thicknesses and incision depths were measured by optical coherence tomography. The 3.0-mm and 6.0-mm zone central-to-peripheral thickness ratios were calculated. Results: Endothelial perforation occurred only in the Pachy+50 group (n=3, 30%). Central lamella's thickness in Pachy-100, Pachy-50, Pachy-0, and Pachy+50 groups measured 185 ± 42 µm, 122 ± 29 µm, 114 ± 29 µm, and 58 ± 31 µm, respectively (p<0.001). The overall 3.0- and 6.0-mm C/P ratios were 0.97 ± 0.06 and 0.92 ± 0.14, respectively. Preoperative donor characteristics were not correlated with most thickness outcomes. The planned incision depth correlated significantly with most lamella's thickness parameters (p<0.001). The overall thickness of the lamella negatively correlated with the planned incision depth (p<0.001, r=-0.580). The best outcome was found in the Pachy-0 group, as 75% of the lamellae measured <130 µm and there was no endothelial perforation. Conclusions: By using a standardized method of dissection, most manually prepared lamellae presented a planar shape. Setting the incision depth to the central corneal thickness did not result in endothelial perforation and a high percentage of ultrathin lamellae was achieved.

RESUMO Objetivo: Comparar quatro profundidades de dissecção manual usadas no preparo de lamelas para transplante endotelial. Métodos: Córneas humanas de treinamento disponibilizadas foram randomizadas em quatro grupos: Pachy-100 (profundidade de incisão = espessura corneana central - margem de segurança de 100 µm), Pachy-50 (margem de segurança de 50 µm), Pachy-0 (sem margem de segurança) e Pachy+50 (profundidade de incisão = espessura corneana central + 50 µm). Todas as lamelas foram dissecadas através um método padronizado e já publicado (Pachy-DSEK). As espessuras das lamelas (centro, zona de 3,0mm e zona de 6,0mm) foram medidas com tomografia de coerência óptica. A razão de espessura centro-periferia foi calculada aos 3,0 e 6,0 mm de diâmetro. Resultados: Perfuração endotelial ocorreu apenas no grupo Pachy+50 (n=3, 30%). A espessura central da lamela nos grupos Pachy-100, Pachy-50, Pachy-0 e Pachy+50 foi de 185 ± 42 µm, 122 ± 29 µm, 114 ± 29 µm, e 58 ± 31 µm, respectivamente (p<0,001). As razões C/P aos 3,0 e 6,0 mm foram de 0,97 ± 0,06 e 0,92 ± 0,14, respectivamente. Os parâmetros de características do doador não se correlacionaram com os resultados de espessura de lamela. A profundidade planejada de incisão se correlacionou com a maioria dos parâmetros de espessura de lamela (p<0,001). A espessura de lamela se correlacionou negativamente com a profundidade planejada da incisão (p<0.001, r=-0,580). O melhor resultado foi observado no grupo Pachy-0, em que 75% das lamelas mediram abaixo de 130 µm e não houve perfuração endotelial. Conclusão: Através de um método padronizado de dissecção, a maioria das lamelas endoteliais apresentou uma configuração planar. O planejamento de profundidade de incisão igual à espessura corneana central resultou em alta porcentagem de lamelas ultrafinas sem ocorrência de perfuração.

El endotelio sano y su disfunción en el riesgo cardiovascular / Healthy Endothelium and its Dysfunction in Cardiovascular Risk

Resumen El endotelio es una monocapa formada por células aplanadas llamadas w, que revisten la parte más interna del corazón, los vasos sanguíneos y los linfáticos. Es considerado un órgano que tiene una función de barrera, pero además se encarga de regular la permeabilidad y tono vascular, hemostasia, inflamación y angiogénesis. Esta revisión se centra sobre todo en las generalidades del endotelio vascular sano y su disfunción. Se analizan los conceptos de activación y disfunción, en donde la activación se considera como un proceso autolimitado, indispensable para la hemostasia y la inflamación. La disfunción endotelial, en cambio, es un proceso patológico, de mayor duración y que se presenta cuando el endotelio ya no puede autorregularse y cambia a un fenotipo proinflamatorio y protrombótico permanente. Esta disfunción es el primer cambio que lleva a la ateroesclerosis y al aumento del riesgo cardiovascular, por esta razón se revisan los principales biomarcadores de disfunción endotelial y riesgo cardiovascular. A medida que se avance en el conocimiento básico del endotelio y su disfunción, será posible diseñar nuevas medidas preventivas o terapéuticas que puedan disminuir dicho riesgo.

Abstract The endothelium is a monolayer of flatten cells named endothelial cells that form the inner layer of the heart, blood, and lymphatic vessels. Its function is not just as a barrier, but it is a regulator of vascular permeability and tone, hemostasis, inflammation, and angiogenesis. This review is about the general aspects of vascular endothelium and endothelial dysfunction that leads to increased vascular risk. Activation and dysfunction are discussed, considering the endothelial activation as a self-limiting process, necessary to promote inflammation and hemostasis. Endothelial dysfunction is a pathological process in which the endothelium loses its ability for self-regulation and acquires a prothrombotic and proinflammation phenotype. Endothelial dysfunction is the initial step for atherosclerosis and increased cardiovascular risk, so the main biomarkers of endothelial dysfunction are reviewed. As basic knowledge about endothelium increases, preventive or therapeutic measures can be designed as treatment or prevention the risk of its dysfunction.

Evaluación de la disfunción del endotelio para el pronóstico de riesgo cardiovascular / Evaluation of endothelial dysfunction for cardiovascular risk prediction i: en

Introducción: Las enfermedades cardiovasculares constituyen la primera causa de muerte en el mundo, por lo que la identificación y modificación de los factores de riesgo asociados a ellas constituyen estrategias priorizadas por la Organización Mundial de la Salud. Contar con un modelo de predicción del riesgo cardiovascular enriquecido con la evaluación de la disfunción endotelial influiría positivamente en estas metas. Objetivos: Identificar la presencia de disfunción endotelial en pacientes con enfermedades cardiovasculares o sin estas y determinar la asociación entre ambas. Métodos: Se realizó un estudio observacional y descriptivo, de serie de casos, en el Centro de Cardiología y Cirugía Cardiovascular del Hospital Provincial Docente Clínico-Quirúrgico Saturnino Lora de Santiago de Cuba, desde enero del 2022 hasta igual mes del 2023, donde se analizaron como variables los factores de riesgo cardiovascular tradicionales y los biomarcadores de disfunción endotelial. Secundariamente, se llevó a cabo un estudio analítico de casos y controles en el cual se aplicó la regresión logística binaria multivariada. Resultados: Se confirmó la presencia de disfunción endotelial asociada a la aparición de las enfermedades cardiovasculares, lo que se evaluó a través del índice de vasodilatación, mediado por el flujo de la arteria braquial y las concentraciones plasmáticas de fibrinógeno. Conclusiones: Las características epidemiológicas y clínicas de los pacientes con enfermedades cardiovasculares o sin estas no difirieron de lo registrado en la literatura especializada acerca de la base de identificación de los factores de riesgo tradicionales.

Introduction: Cardiovascular diseases constitute the first death cause worldwide, reason why the identification and modification of associated risk factors constitute prioritized strategies by the World Health Organization. To have a prediction model of cardiovascular risk enriched with the evaluation of the endothelial dysfunction would influence positively in these goals. Objectives: To identify the presence of endothelial dysfunction in patients with or without cardiovascular diseases and to determine the association between them. Methods: An observational and descriptive cases series study was carried out in the Cardiology and Cardiovascular Surgery Center at Saturnino Lora Teaching Clinical Surgical Provincial Hospital in Santiago de Cuba, from January, 2022 to the same month, 2023, where the traditional cardiovascular risk factors and endothelial dysfunction biomarkers were analyzed as variables. Secondarily, an analytic case-control study was carried out in which multivariate binary logistic regression was applied. Results: The presence of endothelial dysfunction associated with the onset of cardiovascular diseases was confirmed, what was evaluated through the vasodilatation index, mediated by the brachial artery flow and the fibrinogen plasmatic concentrations. Conclusions: The clinical and epidemiological pattern of patients with or without cardiovascular diseases did not differ from that reported in the specialized literature on the base of the identification of traditional risk factors.

Clinical outcomes of epithelial ingrowth following laser in situ keratomileusis with mechanical debridement and compressed heating air flow: a case series / Resultado visual do tratamento do crescimento epitelial após laser in situ ceratomileuse através da remoção mecânica e aplicação de ar comprimido aquecido: série de casos

ABSTRACT Purpose: To describe the clinical outcomes of manual scraping of epithelial ingrowth followed by compressed heating air flow after laser in situ keratomileusis (LASIK). Methods: We underwent a retrospective, noncomparative, and interventional case series. Twenty eyes of 17 patients were included in this study. Each patient with a history of LASIK underwent epithelial removal with mechanical debridement followed by compressed heating air flow. Our primary outcome was the recurrence of epithelial ingrowth after 3 months of follow-up, while our secondary outcomes were uncorrected distance visual acuity, corrected distance visual acuity, and complications after surgery. Results: Ten patients (58.8%) were male, and eight eyes of seven (41.2%) patients underwent primary LASIK surgery, while12 eyes of 10 patients had flap-lift retreatment LASIK; sixteen eyes (80.0%) underwent mechanical microkeratome LASIK and four (20.0%) underwent femtosecond laser-assisted LASIK. Mean age at surgical removal of epithelial ingrowth was 37.0 years ± 9.3 years (range 24 to 55 years). There was recurrence of ingrowth in two eyes (10%) after 3 months of follow-up. The mean corrected distance visual acuity of patients before surgery was 0.07 ± 0.09 logMAR, and after the last follow-up was 0.02 ± 0.04 logMAR (p=0.06). The odds ratio of presenting with epithelial ingrowth after LASIK enhancement compared to primary LASIK was 29.41. Conclusion: Manual scraping followed by compressed heating air flow is a safe and effective treatment of clinically significant epithelial ingrowth after LASIK. At the last follow-up, no eye lost any line in corrected distance visual acuity.

RESUMO Objetivo: Descrever os resultados clínicos do tratamento do crescimento epitelial através da técnica de remoção manual seguido da utilização de um compressor de ar comprimido aquecido após a cirurgia de laser in situ keratomileusis (LASIK). Métodos: Vinte olhos de 17 pacientes foram incluídos no estudo. Cada paciente havia sido submetido a cirurgia de LASIK com presença de crescimento epitelial e foi submetido a tratamento cirúrgico para sua retirada. O objetivo primário foi identificar a presença de crescimento epitelial recorrente ao final de 3 meses de seguimento. Os objetivos secundários foram as medidas de acuidade visual sem correção, acuidade visual com correção, e complicações pós-operatórias. Resultados: Dez pacientes (58,8%) eram homens e 7 mulheres. Oito olhos de sete (41,2%) pacientes apresentavam cirurgia de LASIK primária e 12 olhos de 10 pacientes tinham cirurgia de LASIK com retratamento; dezesseis olhos (80%) utilizaram microcerátomo manual e quatro (20%) laser de femtosegundo. A média de idade no momento da cirurgia de remoção do epitélio era de 37,0 anos ± 9,3 (DP) (variando de 24 a 55 anos). Ocorreu recidiva do crescimento epithelial em dois olhos (10%) após 3 meses de seguimento. A acuidade visual sem correção antes da cirurgia era de 0,07 ± 0,09 logMAR, e após a cirurgia passou para 0,02 ± 0,04 logMAR (p=0,06). A chance (odds ration) de aparecimento do crescimento epithelial após uma reoperação de LASIK é 29,41 vezes maior do que no LASIK primário. Conclusão: A técnica de remoção epitelial manual seguida da utilização de ar comprimido aquecido é segura e efetiva no tratamento do crescimento epitelial após LASIK. Ao final do último acompanhamento, nenhum olho apresentou perda de linhas de visão.

In vivo confocal microscopic observations of 49 patients with posterior polymorphous corneal dystrophy / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To explore the imaging features of 49 patients with posterior polymorphous corneal dystrophy(PPCD)by in vivo confocal microscopy(IVCM).METHODS: Retrospective case series study. A total of 49 patients(86 eyes), including 32 males and 17 females diagnosed with PPCD between January 2013 and January 2021 were collected. The mean age was 42.5±22.9 years. All patients were scanned by IVCM to analyze the density of corneal endothelial cells and described IVCM characteristics of different types of PPCD.RESULTS: The number of endothelial cells in the lesion area of all patients was lower than that in the peripheral area. Under IVCM, 44 eyes(51%)were categorized into type 1 PPCD(vesicular lesions), characterized by single or multiple, central round or irregular crater-like lesion on paracentral corneal endothelial layer; 16 eyes(19%)were categorized into type 2 PPCD(band lesions), which displayed curved and raised edge with scattered or banded-distributed gutta-like lesion between edges. Type 3 PPCD(diffuse lesion)were in 26 eyes(30%), which showed that endothelial cells were missing in many areas. The blurred images of endothelium in most areas featured with spikes lined in a streak, and the clear images in some areas featured with a band lesions. Two patients were followed up for 4-5a. The IVCM images showed different lesions, including the decrease of central corneal endothelial cell density and the iron deposit in the corneal epithelium, etc.CONCLUSION: IVCM is able to scan the characteristic microstructural alterations at the level of endothelium and Descemet membrane in patients with PPCD, and provide an effective image diagnosis for PPCD.

Pharmacodynamic Material Basis of Astragali Radix and Angelicae Sinensis Radix Drug Pair in Supplementing Qi and Activating Blood Circulation / 中国实验方剂学杂志

ObjectiveTo evaluate the effect of Astragali Radix （AR）-Angelicae Sinensis Radix （ASR） drug pair on supplementing Qi and activating blood circulation in rats with Qi deficiency and blood stasis and provide a theoretical basis for clinical rational medication and identification and quality control of compound pharmacodynamic substances from the three aspects of characteristic map， identification of pharmacodynamic substances， and comparison of blood components. MethodHigh-performance liquid chromatography （HPLC） was employed to establish the fingerprint of AR∶ASR （3∶1）， and ultra-high performance liquid chromatography-Q-Exactive Orbitrap-mass spectrometry （UPLC-Q-Exactive Orbitrap-MS） was employed to analyze the ingredients of the decoction. Adult male Wistar rats with SPF grades were selected and randomly divided into a blank group， a model group， a 3∶1 group， and a 5∶1 group. The rat model of Qi deficiency and blood stasis syndrome was prepared by controlling food intake and swimming in cold water every day. In parallel， each group was given medicine （or water） once a day. The dose of drug groups was 10.2 g∙kg-1， and the model group and blank group were given the same amount of distilled water for 15 d. Animal behavior， body weight， whole blood and plasma viscosity， thymus index， spleen index， the levels of adenosine triphosphate （ATP）， adenosine diphosphate（ADP）， von willebrand factor （vWF）， and ATP/ADP value in serum of rats were recorded. The morphology of vascular endothelium was observed by hematoxylin-eosin （HE） staining and scanning electron microscopy. UPLC-Q-Exactive Orbitrap-MS was used to analyze prototype and metabolic components in serum. ResultThe fingerprint of AR-ASR drug pair （AR-ASR 3∶1） was established. UPLC-Q-Exactive Orbitrap MS identified 49 chemical components in vitro and preliminarily identified 11 prototype components absorbed into blood in vivo. As compared with the blank group， the body mass decreased significantly （P<0.01）， the whole blood （high shear， middle shear， and low shear） viscosity and plasma viscosity were significantly increased （P<0.05， P<0.01）， the thymus index and spleen index decreased significantly （P<0.05， P<0.01）， serum ATP content decreased significantly （P<0.01）， ADP content increased significantly （P<0.01）， ATP/ADP value decreased significantly （P<0.01）， and vWF content increased significantly （P<0.01）. The results of HE staining and scanning electron microscopy showed that the vessels were partially damaged， showing the structural disorder of the intima， the bulge， defect， and roughness of the endothelium， and the obvious cell adhesion and migration in the model group. As compared with the model group， the body mass also increased significantly （P<0.01）. The results of whole blood and plasma viscosity showed that the whole blood low shear viscosity was significantly decreased in the 3∶1 group （P<0.05）. The results of thymus index and spleen index showed that 5∶1 group significantly increased the thymus index of rats （P<0.05）. The results of serum ATP and ADP levels showed that the 5∶1 group had more significant effects on ATP and ADP levels （P<0.05）， and both groups significantly reduced ATP/ADP values （P<0.01）. The results of serum vWF level showed that the vWF content in the 3∶1 group decreased significantly （P<0.05）. The results of HE staining and scanning electronic microscopy showed that the damage of vascular endothelium was improved in the treatment group and the structure of intima was neat. ConclusionAR-ASR drug pair can improve the macro and micro indexes of rats with qi deficiency and blood stasis in the 3∶1 and 5∶1 groups. Overall， the 5∶1 ratio has a better effect on supplementing Qi but 3∶1 ratio has a better effect on promoting blood circulation.