Effects of chronic smoking on central corneal thickness,endothelial cell,and dry eye parameters

Objective: To evaluate the effect of chronic cigarette smoking on dry eye parameters, endothelial cells, and corneal thickness.

Design: Prospective cross-sectional case series.

Methods: In this cross-sectional study, 49 eyes of 49 chronic smokers (smoker group) and 53 eyes of 53 age-matched, healthy non-smokers (non-smoker group) were enrolled. All participants underwent measurements of tear breakup time (TBUT), central corneal thickness (CCT) measurements with contact pachymeter and the Schirmer test with anesthesia. Corneal endothelial cells were evaluated by non-contact specular microscopy and photographed for analysis of cell density and hexagonality and the coefficient of variation in cell size.

Results: The mean Schirmer score and TBUT value were significantly lower in the smoker group compared to the non-smoker group (p?=?0.015) and p?p?>?0.05). However, a lower percentage of endothelial hexagonal cells were observed in smokers than non-smokers (p?Discussion and conclusion: Our results suggest that cigarette smoking seems to affect the Schirmer score, TBUT value, and hexagonal cells of the corneal endothelium.     

受体血清和Hanks液保存兔角膜植片的内皮细胞比较

目的：评价血清对角膜植片保存的意义。方法：对新鲜兔角膜在受体血清和Hanks液中分别进行24h和72h保存后，各组的平均内皮细胞密度及内皮细胞存活率等指标进行比较。结果：血清74h保存组的植片内皮细胞各项指标均优于Hanks液72h保存组。结论：血清有着维护角膜内皮细胞结构和功能的作用。     

双重粘弹剂在伴有2型糖尿病的白内障患者超声乳化手术中对角膜内皮的保护作用

目的 探讨伴有2型糖尿病的白内障患者在超声乳化手术中使用双重粘弹剂, 对角膜内皮的保护作用。方法 将56例伴有2型糖尿病的白内障的患者(60眼)随机分为两组, 试验组于术中使用粘弹剂Duovisc, 对照组术中使用粘弹剂透明质酸钠。分别检测两组患者术前、术后3天、1个月、3个月的中央角膜厚度与角膜内皮细胞密度。结果 患者术后3天、1个月、3个月的角膜内皮细胞密度均低于术前, 差异有统计学意义(P<0.05);术后3天、1个月对照组中央角膜厚度显著高于试验组, 差异有统计学意义(P<0.05)。结论 伴有糖尿病的白内障患者在超声乳化手术中使用双重粘弹剂可有效保护角膜内皮细胞。     

The effect and safety of intravitreal injection of ranibizumab and bevacizumab on the corneal endothelium in the treatment of diabetic macular edema

Objective: To investigate the effect and safety of intravitreal injection (IVI) of bevacizumab and ranibizumab on corneal endothelial cell count and morphology in patients with diabetic macular edema.

Materials and methods: A total of 60 eyes from 60 consecutive patients who received 0.5?mg/0.05?ml IVIs of bevacizumab (n?=?30, IVB group) or 1.25?mg/0.05?ml ranibizumab (n?=?30, IVR group) for three consecutive months were investigated prospectively. Specular microscopy was performed to evaluate endothelial cell count, the percentage of hexagonal cells (pleomorphism), and the coefficient of variation of the cell size (polymegathism); optical biometry was performed to evaluate central corneal thickness. Results before injection and 1 month after the first and third injections were compared.

Results: The groups were matched for age (p?=?0.11) and gender (p?=?0.32). There was no significant difference in endothelial cell count (IVB group, p?=?0.66; IVR group, p?=?0.74), pleomorphism (IVB group, p?=?0.44; IVR group, p?=?0.88) and polymegathism (IVB group, p?=?0.21; IVR group, p?=?0.24) before injection or 1 month after the first and third injections. There was also no difference in central corneal thickness (IVB group, p?=?0.15; IVR group, p?=?0.58) before injection or 1 month after the first and third injections.

Conclusion: Monthly 1.25?mg/0.05?ml IVIs of bevacizumab or 0.5?mg/0.05?ml of ranibizumab for three consecutive months in the treatment of diabetic macular edema does not affect corneal morphology and has no harmful effects on the endothelium.     

2型糖尿病患者角膜厚度变化临床探讨

目的：探讨2型糖尿病患者角膜厚度，评价其角膜内皮细胞功能。方法：应用Orbscan生物测量系统对2型糖尿病患者35例（70只眼）角膜进行厚度检测，分别计算中央、距中心1．5、2．5、3．5、4．5mm处平均角膜厚度，并以20例正常健康眼角膜作为对照。结果：与对照组比较，2型糖尿病患者各部位角膜厚度均有增加，两者差异有显著性（P〈0．05）。结论：2型糖尿病患者的角膜增厚．提示内皮细胞功能低下。     

白内障超声乳化术对角膜内皮细胞数的影响分析

目的：探讨白内障超声乳化术对角膜内皮细胞数的影响。方法：选取2008年1月-2010年12月于本院进行治疗的64例白内障患者为研究对象,将其根据手术方式分为对照组（非超声乳化小切口手术组）32例和观察组（超声乳化术组）32例,后将两组患者术前及术后3个月的角膜内皮细胞数、密度及六角形细胞比例进行统计及比较。结果：术后观察组角膜内皮细胞数、密度及六角形细胞比例均高于对照组,P均〈0．05,差异均有统计学意义。结论：白内障超声乳化术对角膜内皮细胞数的不良影响要小于非超声乳化小切口手术。     

无动物成分器官培养法保存角膜内皮细胞活性的实验研究

目的探讨无动物成分(ACF)器官培养保存角膜内皮细胞活性方面的效果。方法将Wistar大鼠角膜16只密闭保存于ACF中4周,葡聚糖T500脱水2 d,作为ACF组。以常规含2%胎牛血清(FBS)的MEM基础培养基组作为对照组。保存结束,计数角膜内皮细胞密度(ECD),免疫组织化学染色法和RT-PCR法检测大鼠角膜内皮层中胞质紧密粘连蛋白1(ZO-1)的表达水平,以评估内皮细胞活性和细胞间紧密连接的屏障功能。结果保存结束后,对照组大鼠角膜的ECD值仅为(1 875±162)个/mm2,ACF组则高达(2 143±169)个/mm2,两组间比较差异显著(P=0.006)。角膜冰冻切片显示ZO-1在保存后大鼠内皮层成功表达。RT-PCR检测表明ZO-1mRNA在ACF组的表达水平高于对照组。结论 ACF器官培养法保存角膜可以更好地保持内皮细胞的活性和紧密连接的屏障功能,在无血清器官培养保存角膜方面具有良好的应用前景。     

Non-treatment-seeking heavy drinkers: effects of chronic cigarette smoking on brain structure

We previously reported [Cardenas, V.A., Studholme, C., Meyerhoff, D.J., Song, E., Weiner, M.W., 2005. Chronic active heavy drinking and family history of problem drinking modulate regional brain tissue volumes. Psychiatry Res. 138, 115-130] that non-treatment-seeking, active heavy drinkers (HD) demonstrated smaller regional neocortical gray matter volumes compared to light drinking controls; however, the potential effects of chronic cigarette smoking on regional brain volumes were not addressed. The goal of this retrospective analysis was to determine if chronic smoking affected brain structure in the non-treatment-seeking heavy drinking sample from our earlier report (i.e., Cardenas et al., 2005). Regional volumetric comparisons were made among age-matched smoking HD (n=17), non-smoking HD (n=16), and non-smoking light drinkers (nsLD; n=20) from our original sample. Quantitative volumetric measures of neocortical gray matter (GM), white matter (WM), subcortical structures, and cerebral spinal fluid (CSF) were derived from high-resolution magnetic resonance imaging. Smoking HD demonstrated smaller volumes than nsLD in the frontal, parietal, temporal GM, and for total neocortical GM. Smoking HD also demonstrated smaller temporal and total GM volumes than non-smoking HD. Non-smoking HD and nsLD did not differ significantly on GM volumes. Further, the three groups did not differ on lobar WM, subcortical structures or regional CSF volumes. These retrospective analyses indicate neocortical GM volume reductions in non-treatment-seeking smoking HD, but not in non-smoking HD, which are consistent with our studies in recently detoxified treatment-seeking alcohol-dependent samples.     

The effect of acute exposure to coarse particulate matter air pollution in a rural location on circulating endothelial progenitor cells: results from a randomized controlled study

Abstract

Context: Fine particulate matter (PM) air pollution has been associated with alterations in circulating endothelial progenitor cell (EPC) levels, which may be one mechanism whereby exposures promote cardiovascular diseases. However, the impact of coarse PM on EPCs is unknown.

Objective: We aimed to determine the effect of acute exposure to coarse concentrated ambient particles (CAP) on circulating EPC levels.

Methods: Thirty-two adults (25.9?±?6.6 years) were exposed to coarse CAP (76.2?±?51.5?μg?m^?3) in a rural location and filtered air (FA) for 2 h in a randomized double-blind crossover study. Peripheral venous blood was collected 2 and 20 h post-exposures for circulating EPC (n?=?21), white blood cell (n?=?24) and vascular endothelial growth factor (VEGF) (n?=?16–19) levels. The changes between exposures were compared by matched Wilcoxon signed-rank tests.

Results: Circulating EPC levels were elevated 2 [108.29 (6.24–249.71) EPC?mL^?1; median (25th–75th percentiles), p?=?0.052] and 20 h [106.86 (52.91–278.35) EPC?mL^?1, p?=?0.008] post-CAP exposure compared to the same time points following FA [38.47 (0.00–84.83) and 50.16 (0.00–104.79) EPC?mL^?1]. VEGF and white blood cell (WBC) levels did not differ between exposures.

Conclusions: Brief inhalation of coarse PM from a rural location elicited an increase in EPCs that persisted for at least 20 h. The underlying mechanism responsible may reflect a systemic reaction to an acute “endothelial injury” and/or a circulating EPC response to sympathetic nervous system activation.     

Effects of cigarette smoking on psychopathology scores in patients with schizophrenia: An experimental study

Cigarette smoking and/or nicotine administration have been shown to transiently ameliorate several psychophysiological deficits in patients with schizophrenia such as indicators of deficient sensory gating and attention, but acute effects of smoking on positive and negative symptoms in schizophrenia have not been evaluated in experimental paradigms. The current study assessed whether smoking of cigarettes, after 6–12 h abstinence, transiently alters the expression of negative and/or positive symptoms in patients with schizophrenia who have a history of regular smoking. In a double‐blind, placebo controlled study patients with schizophrenia participated in two sessions in which they smoked either cigarettes moderately high in nicotine content or denicotinized cigarettes. They were interviewed pre‐and postsmoking to obtain ratings of PANSS and SANS scales, and had blood pressure and pulse serially recorded before and after smoking. Pulse rate and blood pressure were slightly higher after smoking in the high nicotine cigarette session. Negative symptom scores on both scales were significantly lower after cigarette smoking compared to same‐day predrug baseline, but there were no differences in active versus denicotinized cigarette drug effects. These results suggest that acute smoking of cigarettes reduces negative symptoms in patients with schizophrenia in this experimental paradigm. Future work needs to identify the mechanism responsible for this behavioral effect.     

温石棉与吸烟联合作用于人胚肺细胞时对活性氧产生的影响

为探讨温石棉（ＣＨ）与吸烟联合作用于人胚肺（ＨＥＬ）细胞时对反应体系中活性氧产生的影响，本研究采用不同剂量的ＣＨ和香烟烟溶液（ＣＳＳ）单独及联合作用于ＨＥＬ细胞，观察了Ｈ２Ｏ２、Ｏ—·２和·ＯＨ的产生情况。结果表明，ＣＨ和ＣＳＳ单独与ＨＥＬ细胞作用２小时，反应体系中Ｈ２Ｏ２的产生均明显升高，当不同剂量的ＣＨ和ＣＳＳ联合作用时Ｈ２Ｏ２的增加呈相加或加强作用；ＣＳＳ与ＨＥＬ细胞单独作用１小时，反应体系中Ｏ—·２也明显升高，当ＣＨ与ＣＳＳ同时作用时对Ｏ—·２的增加呈相加作用；用ＥＳＲ直接测定反应体系中·ＯＨ的产生情况表明ＣＨ和ＣＳＳ可协同增加·ＯＨ的产生。     

阿司匹林对人主动脉血管内皮细胞炎性损伤的保护作用

目的：研究阿司匹林（aspirin,Asp）对脂多糖（lipopolysaccharide,LPS）诱导人主动脉内皮细胞（human aortic endothelial cells,HAECs）损伤的保护作用,并进一步阐明其对一氧化氮合酶（NOS）及血管内皮生长因子（VEGF）及其相关受体信号的调控。方法：LPS建立HAECs损伤模型。苏木精-伊红（HE）染色观察细胞形态;MTT法、划痕实验分析HAECs损伤修复能力;ELISA测定一氧化氮（NO）含量;Western blot检测内皮型一氧化氮合酶（eNOS）、诱导型一氧化氮合酶（iNOS）、VEGF和血管内皮生长因子受体-2（VEGFR-2）蛋白表达。结果：给药12 h后Asp明显改善LPS（5 mg·L^-1）导致的细胞损伤、提高修复能力（P<0.05）,并上调NO分泌量及VEGF、VEGFR-2的蛋白表达（P<0.01）;升高eNOS蛋白的表达（P<0.01）。而给药24 h后阿司匹林显著下调LPS导致的NO分泌量及iNOS、VEGF、VEGFR-2的蛋白表达升高,同时升高eNOS蛋白的表达（P<0.01）。结论：阿司匹林对LPS诱导的血管内皮细胞炎性损伤的保护作用与调节NOS/NO和VEGF及其受体的动态平衡密切相关。     

日本刺参酸性粘多糖、皂苷和胶原蛋白多肽对血管内皮细胞的保护作用

目的研究日本刺参酸性粘多糖、皂苷和胶原多肽对血管内皮细胞的保护作用。方法采用ox-LDL建立体外培养的人脐静脉血管内皮细胞株(ECV304)损伤模型。MTT法测定血管内皮细胞的增殖活性;硝酸还原酶法测定血管内皮细胞一氧化氮合酶(NOS)活力和NO释放量;TBA法测定细胞内MDA含量;DNA-Ladder法检测血管内皮细胞的凋亡。结果不同浓度的酸性粘多糖、胶原蛋白多肽和低浓度的皂苷能明显抑制ox-LDL对血管内皮细胞的损伤,促进血管内皮细胞增殖(P<0.05,P<0.01);降低细胞内MDA含量(P<0.05,P<0.01);拮抗ox-LDL引起的血管内皮细胞凋亡。酸性粘多糖和胶原蛋白多肽还能明显促进血管内皮细胞NO释放量(P<0.05,P<0.01),提高细胞NOS活力(P<0.05,P<0.01)。结论日本刺参酸性粘多糖、皂苷和胶原蛋白多肽对血管内皮细胞的脂质过氧化物损伤均具有明显的保护作用。     

Acute effects of short-chain alkylglycerols on blood-brain barrier properties of cultured brain endothelial cells

Background and Purpose

The blood-brain barrier (BBB) restricts drug penetration to the brain preventing effective treatment of patients suffering from brain tumours. Intra-arterial injection of short-chain alkylglycerols (AGs) opens the BBB and increases delivery of molecules to rodent brain parenchyma in vivo. The mechanism underlying AG-mediated modification of BBB permeability is still unknown. Here, we have tested the effects of AGs on barrier properties of cultured brain microvascular endothelial cells.

Experimental Approach

The effects of two AGs, 1-O-pentylglycerol and 2-O-hexyldiglycerol were examined using an in vitro BBB model consisting of primary cultures of rat brain endothelial cells, co-cultured with rat cerebral glial cells. Integrity of the paracellular, tight junction-based, permeation route was analysed by functional assays, immunostaining for junctional proteins, freeze-fracture electron microscopy, and analysis of claudin-claudin trans-interactions.

Key Results

AG treatment (5 min) reversibly reduced transendothelial electrical resistance and increased BBB permeability for fluorescein accompanied by changes in cell morphology and immunostaining for claudin-5 and β-catenin. These short-term changes were not accompanied by alterations of inter-endothelial tight junction strand complexity or the trans-interaction of claudin-5.

Conclusion and Implications

AG-mediated increase in brain endothelial paracellular permeability was short, reversible and did not affect tight junction strand complexity. Redistribution of junctional proteins and alterations in the cell shape indicate the involvement of the cytoskeleton in the action of AGs. These data confirm the results from in vivo studies in rodents characterizing AGs as adjuvants that transiently open the BBB.     

Effects of puerarin on number and activity of endothelial progenitor cells from peripheral blood

AIM: To investigate whether puerarin can augment endothelial progenitor cells (EPCs) numbers, promote EPC proliferative, migratory, adhesive, and in vitro vasculogenesis capacity. METHODS: EPCs were characterized as adherent cells by double staining of DiLDL-uptake and lectin binding under a laser scanning confocal microscope. Expression of KDR, VEGFR-2, and AC 133 was detected by flow cytometry. EPCs proliferation, migration and in vitro vasculogenesis were determined with MTT assay, modified Boyden chamber assay, and in vitro vasculogenesis kit, respectively. EPCs adhesive assay was performed by replating those on fibronectin-coated dishes, then adherent cells were counted. RESULTS: Incubation of isolated human MNCs with puerarin 0.1-3 mmol/L increased the number of EPCs, EPC proliferative, migratory, adhesive, and in vitro vasculogenesis capacity in a concentration and time-dependent manner, which reached maximum at 3 mmol/L, 24 h (approximately 1-fold increase, P<0.01). CONCLUSION: Puerarin enhanced     

褪黑素增强内皮细胞抗LPS损伤能力的研究

目的探讨褪黑素对体外培养的内皮细胞抗内毒素损伤的影响。方法体外培养内皮细胞,加入5mg/L脂多糖(LPS)和(或)褪黑素(0.1mmol/L)干预培养24h后,检测细胞乳酸脱氢酶释放率、丙二醛含量;应用RT-PCR检测各组细胞肿瘤坏死因子(TNF-α)转录水平的改变;噻唑蓝(MTT)法检测细胞存活率。结果 LPS刺激24h后,TNF-αmRNA表达量显著升高,细胞存活率下降、而丙二醛含量和乳酸脱氢酶释放率均增高(P<0.05);加用褪黑素后,TNF-αmRNA表达量下降,细胞存活率提高、丙二醛含量和乳酸脱氢酶释放率下降。结论褪黑素可增强体外培养内皮细胞的抗LPS损伤能力     

眼镜蛇毒L-氨基酸氧化酶分离纯化及对血管内皮细胞的抑制作用

目的从眼镜蛇毒中分离纯化L-氨基酸氧化酶(L-a-mino acid oxidase of Naja atra venom,NAV-LAAO),观察其对血管内皮细胞是否有抑制作用。方法应用阳离子交换层析和亲和层析方法分离纯化NAV-LAAO;采用MTT法、Western blot测定caspase及细胞小管形成实验观察NAV-LAAO对人脐静脉内皮细胞(HUVEC)生长和血管生成的影响。结果眼镜蛇毒粗毒经两步层析法得到的LAAO,分子量约为58 ku。NAV-LAAO抑制HUVEC细胞的生长和细胞小管形成,其抑制细胞生长的IC50为21.42 mg.L-1。与对照组相比,LAAO组caspase-3、caspase-8升高。结论应用两步层析法从眼镜蛇毒中分离出的NAV-LAAO具有剂量依赖性的抑制HUVEC细胞的生长和影响细胞小管形成的作用。     

Ocular delivery of cyclosporine A based on glyceryl monooleate/poloxamer 407 liquid crystalline nanoparticles: preparation,characterization, in vitro corneal penetration and ocular irritation

The purpose of this study was to develop an ophthalmic drug delivery system for cyclosporine A (CsA) based on glyceryl monooleate (GMO)/poloxamer 407 liquid crystalline nanoparticles with reduced ocular irritancy and improved corneal penetration. CsA-loaded liquid crystalline nanoparticles were prepared via fragmentation of a bulk GMO/poloxamer 407 cubic phase gel by high-pressure homogenization and characterized. Corneal permeation and retention was evaluated using modified Franz diffusing cells. Intra-corneal transportation was investigated with fluorescein isothiocyanate (FITC)-labeled liquid crystalline nanoparticles. Ocular irritation was then evaluated using the Draize method. The mean particle size of liquid crystalline nanoparticles was 193.5?nm and the entrapment efficiency was 95.11?±?0.67%. A bicontinuous cubic phase of cubic P-type was determined using cryo-transmission electron microscopy (cryo-TEM) observation and small angle X-ray diffraction (SAXD) analysis. A 1.52-fold increase in J_s and a 2.2-fold increase in corneal retention was achieved by liquid crystalline nanoparticles compared with oil solution. In vitro corneal permeation investigated with FITC-labeled liquid crystalline nanoparticles revealed that CsA penetrated across the cornea under the transportation of liquid crystalline nanoparticles. Liquid crystalline nanoparticles exhibited excellent ocular tolerance in the ocular irritation test. This low-irritant vehicle based on liquid crystalline nanoparticles might be a promising system for effective ocular CsA delivery.