Effects of moxibustion at 46°C on blood lipids and related indicators of thoracic aortic endothelium in a hyperlipidemia rat model

Objective:To observe the effects of moxibustion at different temperatures(38℃,46℃)on blood lipids,endothelial morphology of the thoracic aorta,serum endothelin-1(ET-1),calcitonin gene-related peptide(CGRP),nitric oxide(NO),and endothelial NO synthase(eNOS)in hyperlipidemic rats.Methods:Using the random number table method,60 Sprague Dawley rats were randomly and evenly divided into blank,model,38℃-moxibustion,and 46℃-moxibustion groups.Rats in the 3 experimental groups were fed a high-fat feed to model hyperlipidemia in rats.Rats in the 38℃-moxibustion and 46℃-moxibustion groups were moxibustion on the Shenque and bilateral Zusanli acupoints for 10 minutes each,once every other day for 4 weeks,at temperatures of 38±1℃ and 46±1℃.After that,rat blood samples were collected to detect blood lipids and ET-1,CGRP,eNOS and NO.Take the endotheal tissue of the thoracic aorta to do HE staining.Results:(1)The serum total cholesterol,triglycerides,and low-density lipoprotein cholesterol of rats in the 46℃-moxibustion group were significantly lower than those in the model and 38℃-moxibustion groups.(2)Revealed by hematoxylin and eosin staining,showed necrosis in the local vascular endothelial cells and mild inflammatory cell infiltration in the tunica adventitia of the hyperlipidemic rats.These endothelial morphologies did not improve significantly after moxibustion at 38℃ but did improve at 46℃.(3)Compared with the blank group,serum ET-1 was significantly higher and serum CGRP,NO,eNOS were significantly lower in the model group.Compared with the model and the 38℃-moxibustion groups,serum ET-1 was significantly lower and serum CGRP,NO,eNOS were significantly higher in 46℃-moxibustion groups.Conclusion:Moxibustion at 46℃ effectively regulated blood lipids,improved the morphology of the vascular endothelium,and protected vascular endothelial function.

Endothelium-derived essential signals involved in pancreas organogenesis

Endothelial cells(ECs) are essential for pancreas differentiation, endocrine specification, and endocrine function. They are also involved in the physiopathology of type 1 and type 2 diabetes. During embryogenesis, aortic ECs provide specific factors that maintain the expression of key genes for pancreas development such as pancreatic and duodenal homeobox-1. Other unknown factors are also important for pancreatic endocrine specification and formation of insulin-producing beta cells. Endocrine precursors proliferate interspersed with ductal cells and exocrine precursors and, at some point of development, these endocrine precursors migrate to pancreatic mesenchyme and start forming the islets of Langerhans. By the end of the gestation and close to birth, these islets contain immature beta cells with the capacity to express vascular endothelial growth factor and therefore to recruit ECs from the surrounding microenvironment. ECs in turn produce factors that are essential to maintain insulin secretion in pancreatic beta cells. Once assembled, a cross talk between endocrine cells and ECs maintain the integrity of islets toward an adequate function during the whole life of the adult individual. This review will focus in the EC role in the differentiation and maturation of pancreatic beta cells during embryogenesis as well as the current knowledge about the involvement of endothelium to derive pancreatic beta cells in vitro from mouse or human pluripotent stem cells.

Changes in aortic endothelium ultrastructure in male rats following castration, replacement with testosterone and administration of 50α-reductase inhibitor

Aim： To investigate the relationship between low androgen level and ultrastructure of vascular endothelium. Methods： Forty-eight male Sprague-Dawley rats were randomly divided into four groups： group A, normal rats with sham castration; group B, castrated rats; group C, castrated rats given testosterone （T） undecanoate; and group D, intact rats treated with 5α-reductase inhibitor. After 10 weeks of treatment or castration, rats in different groups were killed and serum T, free T （FT） and dihydrotestosterone （DHT） were measured. The aortic endothelia were scanned under electron microcopy and the Vascular Endothelium Structure Score （VESS） was computed. Results： Serum T and FT concentrations of rats in group B were significantly lower than those of the other three groups （P 〈 0.01）; DHT concentrations of group D rats were significantly decreased （P 〈 0.01 ） when compared with those of groups A and C. Rats in groups B and D rats （with low androgen levels） had obvious damage to their endothelial surfaces, which appeared crimpled, rough, adhesive and ruptured, and had high destruction of VESS. Conclusion： These results suggest that low concentrations of T and DHT are associated with ultrastructural damage of the aortic endothelia in male rats.

Effect of topical 0.05% cyclosporine A on corneal endothelium in patients with dry eye disease

· AIM: To determine the effect of topical 0.05% cyclosporine A (CsA) on corneal endothelium in patients with dry eye disease. · METHODS: Observational, prospective, case series study. Fifty-five eyes of 29 consecutive patients (9 males and 20 females; median age: 66.8 years, interquartile range: 61 -73.2 years) with moderate -severe dry eye disease were evaluated. All patients were treated with topical 0.05% CsA ophthalmic emulsion twice a day in addition to lubricant eyedrops 5 times a day. The follow- up period was 12 months. Before treatment and at 3 and 12 months post -treatment central corneal specular microscopy was performed. The endothelial cell density (ECD), coefficient of variation of cell size (CoV), and percentage of hexagonal cells (Hex %) were analyzed. ·RESULTS: The median ECDs pre-treatment and at 3 and 12 months post-treatment were 2 352.5/mm 2 (inter- quartile range, 2 178 -2548.5), 2 364/mm 2 (interquartile range, 2 174.25 -2 657.5), and 2 366 cells/mm 2 (inter - quartile range, 2 174.75-2 539.75), respectively (P=0.927, one way ANOVA). The median CoVs pre-treatment and at 3 and 12 months post -treatment were 34.5 (interquartile range, 30 -37), 35 (interquartile range, 30 -38), and 34 (interquartile range, 30.75-38.25), respectively (P=0.7193, one way ANOVA). The median Hex % values pre - treatment and at 3 and 12 months post -treatment were 53 (interquartile range, 47 -58), 54 (interquartile range, 45.75 -59), and 50.5 (interquartile range, 45.75 -58), respectively (P=0.824, one way ANOVA). · CONCLUSION: Treatment of patients with dry eye disease for 12 months with topical 0.05% CsA does not seem to cause substantial changes on corneal endothelium.

Relationship between vascular endothelium and periodontal disease in atherosclerotic lesions: Review article

Inflammation and endothelial dysfunction are linked to the pathogenesis of atherosclerotic disease. Recent studies suggest that periodontal infection and the ensuing increase in the levels of inflammatory markers may be associated with myocardial infarction, peripheral vascular disease and cerebrovascular disease. The present article aimed at reviewing contemporary data on the pathophysiology of vascular endothelium and its association with periodontitis in the scenario of cardiovascular disease.

Differences in energy and corneal endothelium between femtosecond laser-assisted and conventional cataract surgeries：prospective,intraindividual,randomized controlled trial

AIM： To compare intraoperative phacoemulsification parameters and its effect on the corneal endothelium of eyes undergoing femtosecond laser-assisted cataract surgery（FLACS） versus conventional phacoemulsification（CP） cataract surgery.METHODS： Two hundred eyes from one hundred patients were included in a prospective, non-blinded, randomized, controlled, intraindividual clinical study. One hundred eyes underwent FLACS while their one hundred fellow eyes underwent CP. All surgeries were performed using the Victus？ femtosecond laser platform and Infinity？ Vision System phacoemulsification machine. Primary outcome measure was endothelial cell density 6 mo after surgery. Secondary outcome measures included central corneal thickness（CCT）, average cell area, standard deviation, coefficient of variation and hexagonality before surgery and 6 mo after surgery and endothelial cell density loss during this period were also evaluated. Intraoperative efficiency parameters [cumulative dissipated energy（CDE）, total intraocular surgery time, total ultrasound time, total phacoemulsification time, total torsional energy time, total aspiration time, ultrasound energy, torsional amplitude and fluid required during surgery] were also collated. RESULTS： Data from these patients was not considered for analysis. Data from 92 patients were analysed. Postoperative endothelial cell density（cells/mm2） between groups（2211.88±392.49 CP; 2246.31±403.48 FLACS） was not statistically significant（P=0.869）. Total ultrasound time, torsional energy time, CDE and fluid requirements were significantly lower the FLACS group（P〈0.05）. Other parameters did not show statistically significant difference between FLACS and CP.CONCLUSION： FLACS displays significant improvements in phacoemulsification parameters in comparison to CP. There are no significant differences in corneal endothelium measures between FLACS and CP.

VEGF Deficit is Involved in Endothelium Dysfunction in Preeclampsia

This study examined the association of expression of vascular endothelial growth factor(VEGF),a promoter of angiogenesis,with endothelium dysfunction in preeclampsia.The level of VEGF protein and mRNA in the placenta and peripheral blood samples of 30 preeclampsia patients and 30 normotensive pregnant women was measured by immunohistochemistry,real-time reverse transcriptase-polymerase chain reaction(RT-PCR) and enzyme-linked immunosorbent assay(ELISA),respectively.VEGF expression in the human umbilical vein endothelial cells(HUVECs) was blocked by small interfering RNAs(siRNAs).The monolayer barrier function of HUVECs was determined by measuring the fluorescence intensity of BSA that crossed the HUVEC monolayers.The cell proliferation and cell-secreted nitric oxide(NO) level were detected by MTT method and nitrate reductase assay,respectively.The results showed that VEGF was expressed in the syncytiotrophoblasts and endothelial cells of vessels and capillaries in the placenta tissue.The serum level of VEGF in the preeclampsia patients was significantly decreased as compared with that in normal pregnant subjects,although VEGF mRNA expression in the placenta tissue of preeclampsia patients remained still high.Moreover,VEGF deficit could lead to endothelium cell dysfunction,and the administration of VEGF could protect endothelium cells from injury.It was concluded that lack of VEGF contributes to endothelium dysfunction,which may lead to the occurrence and development of preeclampsia.

The Role of Endoplasmic Reticulum Stress-related Apoptosis in Vascular Endothelium Pathogenesis

Vascular endothelium refers to a single layer of endothelial cells that line the inner surface of blood vessels,serving as barriers and transducers between the circulating blood in the lumen and the rest of the vessel wall.Endothelial cells play essential roles in many aspects of vascular biology,such as barrier functions,thrombosis/fibrinolysis,inflammation,angiogenesis,vasoconstriction and vasodilation.

A multiscale model for analyzing the synergy of CS and WSS on the endothelium in straight arteries

A multiscale model was proposed to calculate the circumferential stress （CS） and wall shear stress （WSS） and analyze the effects of global and local factors on the CS, WSS and their synergy on the arterial endothelium in large straight arteries. A parameter pair [Zs, SPA] （defined as the ratio of CS amplitude to WSS amplitude and the phase angle between CS and WSS for different harmonic components, respectively） was proposed to characterize the synergy of CS and WSS. The results demonstrated that the CS or WSS in the large straight arteries is determined by the global factors, i.e. the preloads and the afterloads, and the local factors, i.e. the local mechanical properties and the zero-stress states of arterial walls, whereas the Zs and SPA are primarily determined by the local factors and the afterloads. Because the arterial input impedance has been shown to reflect the physiological and pathological states of whole downstream arterial beds, the stress amplitude ratio Zs and the stress phase difference SPA might be appropriate indices to reflect the influences of the states of whole downstream arterial beds on the local blood flow-dependent phenomena such as angiogenesis, vascular remodeling and atherosgenesis.

Scutellarin Reduces Cerebral Ischemia Reperfusion Injury Involving in Vascular Endothelium Protection and PKG Signal

Flavonoid glycoside scutellarin(SCU)has been widely applied in the treatment of cerebral ischemic diseases in China.In this article,we conducted research on the working mechanisms of SCU in hypoxia reoxygenation(HR)injury of isolated cerebral basilar artery(BA)and erebral ischemia reperfusion(CIR)injury in rat models.In isolated rat BA rings,HR causes endothelial dysfunction(ED)and acetylcholine(ACh)induces endothelium-dependent vasodilation.The myography result showed that SCU(100μM)was able to significantly improve the endothelium-dependent vasodilation induced by Ach.However,SCU did not affect the ACh-induced relaxation in normal BA.Further studies suggested that SCU(10-1000μM)dose-dependently induced relaxation in isolated BA rings which were significantly blocked by the cGMP dependent protein kinase(PKG)inhibitor Rp-8-Br-cGMPs(PKGI-rp,4μM).Pre-incubation with SCU(500μM)reversed the impairment of endothelium-dependent vasodilation induced by HR,but the reversing effect was blocked if PKGI-rp(4μM)was added.The brain slice staining test in rats’model of middle cerebral artery occlusion(MCAO)induced CIR proved that the administration of SCU(45,90 mg/kg,iv)significantly reduced the area of cerebral infarction.The Western blot assay result showed that SCU(45 mg/kg,iv)increased brain PKG activity and PKG protein level after CIR surgery.In conclusion,our findings suggested that SCU possesses the ability of protecting brain cells against CIR injury through vascular endothelium protection and PKG signal.

Transplanting embryonic stem cells onto damaged human corneal endothelium

AIM To investigate whether human embryonic stem cells(hESCs) could be made to attach, grow and differentiate on a human Descemet's membrane(DM).METHODS Spontaneously differentiated hESCs were transferred onto a human corneal button with the endothelial layer removed using ocular sticks. The cells were cultured on a DM for up to 15 d. The genetically engineered hESC line expressed green fluorescent protein, which facilitated identification during the culture experiments, tissue preparation, and analysis. To detect any differentiation into human corneal endothelial-like cells, we analysed the transplanted cells by immunohistochemistry using specific antibodies.RESULTS We found transplanted cells form a single layer of cells with a hexagonal shape in the periphery of the DM. The majority of the cells were negative for octamer-binding transcription factor 4 but positive for paired box 6 protein, sodium potassium adenosine triphosphatase(NaKATPase), and Zona Occludens protein 1. In four of the 18 trials, the transplanted cells were found to express CK3, which indicates that the stem cells differentiated into corneal epithelial cells in these cases. CONCLUSION It is possible to get cells originating from hESCs to become established on a human DM, where they grow and differentiate into corneal endothelial-like cells in vitro.

Two male study groups with adiposity and hypertriglyceridemia were at risk for hypertension and alcohol use declined renal endothelium

Men who attended a Bavarian General Medicine Practice were confidentially invented here. Two male study groups were enrolled to characterize adiposity or hypertriglyceridemia showing that these men were at baseline risk for hypertension [1]. Adverse alcohol consumption mediated dysfunction of renal endothelium as shown here and before [1]. This study found that alcohol use aggravated dyslipidemia, fatty liver disease and critical fasting blood glucose of obese men predicting then late hepatorenal disorders. Overall, two male study groups showed a relevant proportion of men who reported alcohol consumption showing then critical morning urines indicating dysfunction of renal endothelium. The present report looked also at healthy men who reported positive lifestyle behaviour and at men with nonalcohol adiposity and nonalcohol hypertriglyceridemia who then showed normal morning urines indicating functional renal endothelium. Relatively young men at risk were motivated to replace adverse alcohol use by healthy liquids without alcohol and by higher quality of food.

Response of the xenograft endothelium in the concordant xenotransplantation

To investigate the response of the xenograft endothelium in the concordant hamster to rat cardiac xenotransplantation and the mechanism of acute vascular rejection. Methods： The animals were divided into 5 groups randomly： control group, CsA group, splenectomy group, DO splenectomy＋CsA group and D3 splenectomy＋CsA group. Hamster heart was heterotopicaly transplanted to rat abdominal cavity. The graft survival was monitored by palpation of the rat abdominal wall. The histological and ultrastructural changes of the xenogafts were investigated. NF-κB and P-selectin expression in the xenograft were detected. Heme Oxigenase-1 and Bcl-2 expression were also detected in the xenografts of different groups. Results： The mean survival time of the xenografts in control group, CsA group, splenectomy group, DO splenectomy＋CsA group and D3 splenectomy＋CsA group was 3.4±0.55, 3.8±0,45, 6.4± 1.52, 30 and 7.4 ±1.14 days. The rejected graft showed typical acute vascular rejection in control group, CsA group,splenectomy group and D3 splenectomy＋CsA group. Endothelial cells of the rejected xenograft showed dramatic assembly of ribosomes and expansion of the rough endoplasmic reticulum. However, the endothelium of the long-term survived grafts in DO splenectomy＋CsA group showed normal architecture. NF-κB and P-selectin expression were detected in the rejected xenografts. HO-1 expression was observed in the long-term survived xenografts in DO splenectomy＋CsA group. Conclusion： The endothelial cells of the xenograft might be activated during the acute vascular rejection. Expression of HO-1 might inhibit the upregulation of NF-κB and adhesion molecular which decreases the activation of the endotheliuna of the graft.

STUDY IN CYTOTOXICITY OF GENTAMYCIN TO CORNEAL EPITHELIUM AND ENDOTHELIUM IN TISSUE CULTURE

A study in cytotoxicity of gentamyein to tissue-cultured bovine corneal endothelial cells and rabbit corneal epithelial cells is reported. When the cultured cells reached confluence, they were exposed to tissue culture media containing gentamycin for 6 hours. We founl that 0.5% gentamycin caused significant damage to corneal epithelial cells---diffuse plasmolysis, with scattered cell necrosis and 5% loss.While corneal endothelial cells were exposed to 1.6 mg/ml gentamycin, extensive cell loss (approximate- ly 15%) was observed. The damaged cells recovered their normal morphology after 24 hours. When the concentration of gentamycin increased twice, serious damage to cells occured. The area of cell loss reached 40%, and the recovery of cellular morphology Was much slower. This study demonstrates that gentamycin potential cytotoxicity to corneal epithelium and endothelium, suggesting that gentamycin should be rationally used in the treatment of ocular diseases.

Combined antihypertensive effect of GSY and metoprolol,based on endothelium-dependent vasodilatation function

OBJECTIVE To investigate the synergistic effect on dilating blood vessels and anti-hypertension of GYS combined with metoprolol.METHODS ① Spontaneously hypertensive rats(SHR)were administered orally with the vehicle,GSY,metoprolol or GSY combined with metoprolol for 4weeks.Blood pressure,which included SBP,DBP and MBP was measured by a noninvasive method every week.At the end of4 weeks,blood was drawn from the ophthalmic venous plexus to determine blood fat levels(serum TC,TG,LDL-c,HDL-c),liver function(serum ALT,AST),and kidney function(serum BUN,UA and Cr)by the ACCUTE(TBA-40FR)automatic.② The aortae of normal SD rats were prepared and cleaned from periadventitial fat and surrounding connective tissue and cut transversely into 4-mm width rings.To observe different concentration of GYS,metoprolol or GSY combined with metoprolol causing relaxation of the isolated aortic rings precontracted until a stable plateau by noradrenaline(NA)directly or in the presence of eNOS inhibitor L-NAME and cyclooxygenase inhibitor indomethacin(INDO)respectively.③ The concentrations of plasma GSY was determined by the HPLC after rats administered orally with GSY or GSY combined with metoprolol for single-dose.DAS data processing software calculated the pharmacokinetic parameters of GSY.RESULTS There was a significant synergism between GYS and metoprolol in lowering blood pressure and the concentrations of serum TC and LDL-c of SHR.The relaxant effect of GYS combined with metoprolol on the aortic rings precontracted by NA could be attenuated by L-NAME or INDO.The AUC0-tof GSY significantly increased after in conjunction with metoprolol.CONCLUSION GYS combined with metoprolol increases the concentrations of plasma GSY and synergistically lowers blood pressure based on endothelium-dependent vasodilatation function(EDVF).

Evaluation of corneal endothelium after UVA/riboflavin cross-linking in thin keratoconic corneas

Dear Sir,UVA/riboflavin cross-linking （CXL） has been used clinically applied for the treatment of keratoconus and corneal edema via enhancement of corneal stiffness The safety of the corneal endothelium is of prime importance during CXL treatment. In clinical practice, a corneal thickness （CT） of 400um has traditionally been regarded as the minimum treatable thickness, thereby avoiding damage to the corneal endothelium Although CXL has been applied to thinner corneas, using a hypoosmotic solution onto cornea and inducing edema . CXL safety still needs further evaluation because of lower relative concentration of collagen in the hydrated stroma . This study aims to evaluate the changes of corneal endothelial density （ECD） in cases where the CT is 〈400 um before iatrogenic corneal swelling and CXL treatment.

EXPERIMENTAL STUDY ON THE CORNEAL ENDOTHELIUM OF TRAUMATIC CATARACT

The cell morphology of corneal endothelium in 84 mice with experimental traumatic cataract was investigated with stained corneal buttons. In the experimental group, the boundaries between adjacent corneal endothelial cells were significantly distorted, some cell boundaries manifested degenerative changes that led to coalescence of the cells. The mean density and mean area of endothelial cells of the controls showed significant difference from those of the experimental group during the 12 weeks of observ...

Mechanical Regulation of Calcium Response and Trail Formation of Neutrophils on Endothelium

Atherosclerosis or fibrosis and cirrhosis undergo chronic inflammation associated with the adhesion between neutrophils and endothelial cells(ECs)that is mediated by their respective cellular adhesive molecules on stiffened blood vessel wall or extracellular matrix(ECM)under shear flow[1-3].However,the mechanical dependence of calcium flux and trail formation in neutrophils remains unclear yet in these processes.First,the effect of substrate stiffness through ECs on neutrophil calcium spike was quantified when the individual neutrophils adhered to EC monolayer pre-placed onto stiffness-varied polyacrylamide(PA)substrate(5 or 34.88 kPa)or glass surface.Our data indicated that E-/P-selectins and ICAM-1s on HUVECs and b2-integrins,PSGL-1s,and CD44s on neutrophils were all involved in mediating neutrophil calcium spike in a stiffness-dependent manner,in which the increase of substrate stiffness enhanced the calcium intensity and spike number.Such stiffness-dependent calcium response is associated with selectin-induced b2-integrin activation through Syk/Src signaling pathway and the F-actin/myosin II function.Moreover,tension-activated calcium ion channels displayed critical roles in initiating stiffness-dependent calcium spike [4].Second,the trail formation of neutrophils to ECs monolayer pre-placed onto the same PA substrate were also tested under shear flow.Live fluorescence imaging showed that neutrophils are able to form long membrane tethers during migration and subsequently leave behind membranous long-lasting trails under shear,which are enriched in LFA-1,Mac-1,and CD44.Moreover,the formation of the trails was inhibited by blocking LFA-1s and Mac-1s,suggesting an important role forβ2-integrins in the trial formation.The recruitment of monocytes was inhibited when pre-blocking ICAM-1s on flowing monocytes,indicating that the neutrophil’s trails employβ2-integrin-ICAM-1 binding to recruit the monocytes.Intriguingly,both the length and the area of the trails increase with increasing substrate stiffness,resulting in the enhanced monocyte recruitment.Inhibition of actin binding protein Arp2/3 impairs the trail formation and dramatically decreases the neutrophil-dependent monocyte recruitment.These data provide an insight into understanding how stiffening of vascular wall could regulate the calcium flux of adhered neutrophils and thus the immune responses in atherosclerosis.They also imply that local mechanical microenvironment is remodeled with the migration of neutrophils,leaving the trails presented to induce and regulate monocyte recruitment.All the results are meaningful in elucidating the occurrence and development of atherosclerosis or fibrosis from the viewpoint of mechanotransduction and also for the potential intervention of cardiovascular disease progress.

Effects of PAF and PAF antagonist WEB 2170 on relaxation of vascular endothelium after brief deoxygenation and reoxygenation

By means of the tension determination of rat thoracic aortic ring. it was found that PAF depressed the acetylcholine (Ach)-induced relaxation of the rings, having undergone 20 min of deoxygenation followed by 30 min of reoxygenation, to 36. 1% of norepinephine(NE) precontraction. PAF receptor antagonist WEB 2170 markedly improved the Ach-induced relaxation of the brief deoxygenated and reoxygenated rings to 86. 7 % of NE precontraction. The results indicated that PAF may be one of the mediators involved in the endothelium relaxation dysfunction related to brief deoxygenation and reoxygenation, and that PAF antagonist WEB 2170 has the protective effect on endothelium relaxation function.

Hemin restores the impaired endothelium-dependent vasodilatation in diabetic db/db mice and the possible mechanisms involved

Heme oxygenase(HO)is the enzyme that catalyzes the degradation of heme and leads to the formation of carbon monoxide,free iron and biliverdin.Emerging evidence suggests that HO-1,an inducible isoform of HO,can exert vaso-protective effects.However,such vascular benefit