INTRODUCTION

Br inzolamide, a compound structurally related to acetazolamide, was approved for the topical treatment of glaucoma in 1999[1]. Acetazolamide, dorzolamide, and brinzolamide are carbonic anhydrase inhibitors (CAIs),all of which belong to sulfonamide antiglaucomatous drugs.Brinzolamide, which is formulated as a 1% ophthalmic suspension and applied twice daily to lower in traocular pressure (IOP), has fewer systemic side effects than acetazolamide. Brinzolamide is a highly specific, noncompetitive, and reversible carbonic anhydrase II inhibitor[2]. Carbonic anhydrases are zinccontaining enzymes that catalyze the reversible hydration of carbon dioxide and bicarbonate. Inhibition of carbonic anhydrase II in the secretory cells of ciliary processes reduces the rate of aqueous humour formation, consequently lowering IOP[3-4].

Glaucoma, a multifactorial optic neuropathy, is the second leading cause of irreversible blindness worldwide[5]. Although elevated IOP is the principle risk factor, deterioration of ocular perfusion by the vascular system accelerates progression of glaucomatous optic nerve atrophy[6]. In glaucoma, reduced blood flow to the eye has been reported in many studies that used a variety of different techniques to assess ocular blood flow (OBF)[7-9]. Moreover, OBF of glaucoma patients is deficient in the retinal, choroidal, and retrobulbar circulations,and focal ischemia corresponds with areas of glaucomatous visual field loss[6]. Therapeutic agents that reverse this condition could be of great clinical benefit for glaucoma patients. The effect of IOP-lowering medications on OBF remains poorly understood.

CA Is may improve blood perfusion in the eyes in humans.It has been shown that acetazolamide leads to a dilatation of retinal vessel diameters and increases blood flow in the optic nerve head[10]. Similarly, topical application of dorzolamide leads to a significant increase of flow velocities of the retrobulbar vessels as measured by color Doppler imaging[11].The mechanism by which IOP-lowering medications increase OBF remains unknown in glaucoma. It is difficult to determine if the CAI-induced increase in ocular perfusion is secondary to IOP reduction or if it is a primary effect on the ocular vasculature.

There are very few studies that have reported the effect of brinzolamide on OBF. Moreover, the results of these studies have been contradictory due to the heterogeneous study designs and methodologies used to measure OBF. Barnes et al[12] found that brinzolamide significantly increased optic nerve head blood flow in Dutch rabbits. However, Martinez and Sanchez-Salorio[13] suggested that after 5y of treatment,brinzolamide did not augment retrobulbar blood flow when added to timolol in glaucoma patients. Based on these reports,we decided to focus on the relationship between brinzolamide and OBF.

In the current study, we sought to determine the effect of brinzolamide on optic disc blood flow (ODB F) by in vivo laser speckle flowgraphy and to assess the effect and mechanism of brinzolamide on isolated ciliary arteries by an isometric tension recording system.

MA TERIALS AND METHODS

Influence of nitric oxide, prostacyclin, and BKCa channels Isolated rabbit ciliary arteries were pretreated with 100 μmol/L L-NAME (n=7; Figure 5 A), 10 μmol/L indomethac in (n=7;Figure 5B) , or 0.1 μmol/L iberiotoxin (n=7; Figure 5C) 30min before application of the high-K solution. Brinzolamideind uced concentration-dependent relaxations were not altered by these drugs (P＞0.05).

In Vivo Studies

Experimental protocol Te sting of bilateral IOP and ODBF in the left eye of the Dutch rabbits took place every 3h over a period of 24 h at 9:00, 12:00, 15:00, 18:00, 21:00, 24:00,3:00, 6:00, and 9:00. Th e pupils of the rabbits were not dilated pharmacologically during the experimental period.After IOPs were measured, the rabbits were given a 5min rest period, and then ocular circulation was assessed using LS FGNAVITM (Softcare Ltd., Fukuoka, Japan) in a dark room. For each rabbit, the 24h measurements of IOP and ODBF were performed twice. During the first 24h, control measurements were made without any drug treatment. After the rabbits rested for 4d, 1% brinzolamide ophthalmic suspension (Azopt®;Alcon Laboratories, Inc., Ft. Worth, TX, USA) was delivered topically as two drops instilled in the left eye at 9:00, and the measurements were immediately begun. The drug was applied again at 21:00, and the blood flow measurements were continued thereafter.

1.1 研究对象 选择2016年12月-2018年9月在本院自然分娩，在产后42d常规复查时，通过超声检查（经腹和阴超）发现盆底器官位置异常，然后进行产后盆底康复训练的产妇62例纳入研究。患者平均年龄（28.4±0.61)岁，孕次（1.4±0.31)次，产次(2.3±0.41)次，产后平均(105.2±21.54）d。选择同期非妊娠年轻女性45例作为对照组，平均年龄 (24.1±0.76)岁，有性生活史，未孕未育。

Measurement of intraocular pressure Bilateral IOP of rabbits was measured with a rebound tonometer (TonoVet®,Icare Finland Oy, Helsinki, Finland) without any topical anesthetic. In t he first 24h measurements, circadian fluctuations of IOP were recorded. The effects of brinzolamide were assessed four days later when IOP was again recorded for 24h beginning immediately after brinzolamide was instilled into the left eyes.

Figure 1 Map of the optic disc A: A representative map produced by LSFG-NAVITM, the investigator manually set an elliptical area at the outer edge of the optic disc, and LSFG-NAVITM measured MBR of that areas (S: Superior quadrant; T: Temporal quadrant; I: Inferior quadrant; N: Nasal quadrant); B: In the elliptical area, the black part represented the blood vessel and the white part represented the tissue.

Laser speckle flowgraphy In order to assess ODBF, mean blur rate (MBR) of left eye of each rabbit was measured using LSFG-NAVITM. MBR is a parameter that yields a exact measurement of optic disc microcirculation, and shows good correlation with the blood flow parameters measured with other OBF assessment instruments[14-15]. MBR was measured in arbitrary units that make comparison in the same site in the same eye practical. Therefore, the investigator manually determined the measurement area of each rabbit’s optic disk and saved it before measuring MBR (Figure 1A). MBRs of three areas of the optic disc were measured: the average MBR over the vessel area (MBR-V), the average MBR over the tissue area (MBR-T), and the average MBR over the entire optic disk (MBR-A; Figure 1B). All rabbits were examined by one experienced investigator. Detailed methods for measurement of MBR by LSFG-NAVITM have been described by previous paper[16].

Ex Vivo Studies

Isolation of the ocular ciliary arteries Rabbits (weight 2-3 kg)were euthanized by intravenous injection of excess sodium pentobarbital (Abbott, North Chicago, IL, USA). Then, their eyes were immediately removed to ensure that the optic nerve attached to the eye is as long as possible. The eyes were placed in a Krebs solution bubbled with 95% O2 and 5% CO2. Our previous article has described in detail the composition of Krebs solution[17]. Under a dissecting microscope, the ciliary artery segments (length: 3-4 mm) and surrounding connective tissue were separated and cut off from the eyes.

综上所述，形态学、流式细胞术以及免疫组化在检测多发性骨髓瘤患者中均具有较高的应用价值，可按照患者的具体情况，将三者联合使用以提高临床阳性诊断率和疗效监测。

这样临时接到甲方通知，并在最短时间里提供精细准确的解释结论和报告，对李淑荣来说并不是第一次。近五年来，川东北地区页岩气、煤层气等非常规油气勘探不断深化，作为测井公司“绍霞团队”技术带头人之一、页岩油气藏创新分团队的负责人，李淑荣肩负着四川盆地及周缘地区测井解释及方法研究的重任，不能有丝毫松懈。

Isometric tension recording Isolated vascular segments(2 mm in length) were mounted in the chamber of Myograph System® (JP Trading, Aarhus, Denmark). Our previous reports have described in detail the procedure for mounting the ciliary arteries[18-19]. After the equilibration period, contractions of the ciliary artery were evoked by a high-K solution and lasted for 20min. Then, 1 μmol/L carbachol was added to induce relaxation of ciliary artery. If high-K-induced contraction was less than 5 mmol/L or carbachol-induced relaxation was less than 1 mmol/L, such vascular segments were excluded from this study. After verifying the arterial responsiveness to high-K and carbachol, the medium in the chamber was replaced by Krebs solution.

Fluctuation of intraocular pressure In the first 24h measurements, analysis of 10 rabbits indicated that the peak IOP occurred at approximately 21:00. There was no significant difference between right and left eyes for any of the measurements (P＞0.05). Instillatio n of brinzolamide reduced the left IOP at all periods except in the first on e immediately after the drug was delivered (P ＜0.05; Figure 2A). Interestingly,with the exception of immediately after the instillation and at 12:00, 15:00, and 3:00, the IOPs in the right eyes of rabbits that received the brinzolamide in the left eye were also significantly decreased compared to the control IOPs taken four days earlier(Figure 2B). Nevertheless, the IOPs in the right eyes were significantly higher than in the left eyes except immediately after instillation (Figure 2C).

Figure 2 Effects of brinzolamide on rabbit IOP A: The left IOP decreased at all times except the first, which was taken immediately after instillation; B: The right IOP also decreased except at 9:00 (immediately after instillation), 12:00, 15:00 and 3:00; C: The right IOP was higher than the left at all times except immediately after instillation. Brinzolamide was instilled into the left eyes only. aP＜0.05.

根据前文分析，当线缆两端固定点间的距离为最大或最小时，为了尽量减小线缆固定点附近受到的额外弯曲应力，线缆的固定接头需向轨面方向倾斜，倾斜角度根据线缆变换幅度来确定。图4中测得电缆的变化幅度夹角约为70°，工程设计中，该变化幅度夹角取一半，使线缆两端固定点间的距离为最大或最小时，线缆的变化幅度一致，受力形变均衡，如图4 b)所示，此处忽略线缆固定点的变化对线缆最低点的影响。

Brinzolamide not only decreased IOP, but it also augmented ODBF after topical application. This finding is consistent with another study indicating that brinzolamide significantly increased optic nerve head blood flow in Dutch rabbits[12].Similarly, brinzolamide increased retinal blood flow as measured by confocal scanning laser Doppler flowmetry in glaucoma patients[23]. To achieve its effect on OBF, brinzolamide must reach the retina and optic nerve at therapeutically effective concentrations after topical application. However, Martinez and Sanchez-Salorio[13] suggested that after 5y of treatment,brinzolamide did not augment retrobulbar blood flow when added to timolol, a beta-blocker drug, in glaucoma patients.Whether the contradictory results between their study and our study is related to the different areas of measurements in the eye and/or to the different techniques used for assessment of OBF is unclear. From our results, it is possible that glaucoma patients, when treated with topically administered brinzolamide, may not only benefit from the IOP-lowering effect, but also an increase in OBF.

Drugs The following drugs were used: brinzolamide and dorzolamide (Tokyo Chemical Industry, Tokyo, Japan),acetazolamide, indomethacin, iberiotoxin, histamine, L-NAME and carbachol (Sigma, St. Louis, MO, USA). The concentration of these drugs referred to the molarity in the myograph chambers.

故乡小院里的故事当然远远不止这些，还有战斗力超群能打败恶狗的大白鹅“将军”，还有主意多多爱偷吃的小羊“白白”，这里简直就是小动物们的乐园啊！

Statistical Analysis Mean±standard deviations were used to express the measured results and ‘n’ represented the number of studied vessel segments unless specifically indicated.Unpaired two-tailed t-test was performed to analyze the statistical differences between the values. One-way analysis of variance was used to determine differences between these concentration-response curves. Statistical significant was set at probability values less than 0.05.

RESULTS

In Vivo Studies

涉身的含义是什么？莱考夫涉身心智观中的“‘涉身’是指人类‘依据生物性的能力以及身体与社会经验在外部环境中运作’，‘心智’则包括理性与概念，因此心智涉身性就是指人类理性和概念的建构都是涉身的，它们通过人类身体、大脑以及与外部世界的互动式运作而形成。”[3]30

Increase of optic disc blood flow induced by brinzolamide In contrast to the circadian fluctuation of IOP, a nocturnal trough of MBR appeared, with a nadir at approximately 21:00(Figure 3). Compared to control measurements, MBR-V(Figure 3A) was greater at 18:00 and 21:00 (P＜0.05), and MBR-T (Figure 3B) and MBR-A (Figure 3C) were greater at 18:00 (P＜0.05) after instillation of brinzolamide.

Ex Vivo Studies

Relaxation induced by brinzolamide, dorzolamide and acetazolamide For rabbit ciliary arteries pre-contracted with the high-K solution, with the exception of acetazolamide,brinzolamide and dorzolamide induced con centration-dependent relaxation (Figure 4A). For dorzolamide (Figure 4C), and brinzolamide (Figure 4D), the relaxations achieved at 1 mmol/L wer e 27.7%±5.2% ( n=26) and 46.1%±9% (n=21), respectively.

Animals This study was approved by the Animal Care Committee of Akita University, and was adherence to the ARVO Statement for the Use of Animals in Ophthalmic and Vision Research. Ten male Dutch rabbits (weight 1.2-1.5 kg) were used to measure ODBF in vivo. Twenty-four male Japanese white rabbits (weight 2-3 kg) were killed, the ciliary arteries were removed and their isometric tension measured ex vivo.Al l rabbits were kept in a specialized animal center, and standard feed and water were supplied on demand.

Prior to testing the vasodilating effect of brinzolamide,dorzolamide, or acetazolamide, the isolated ciliary arteries were re-contracted with the high-K solution and maintained for 20min. Then, these drugs (10 μmol/L-1 mmol/L) were applied cumulatively every 10min. The amplitudes of high-K-induced contractions were defined as 100%.

Figure 3 Effects of brinzolamide on ODBF Brinzolamide was instilled into the left eye of each rabbit. The MBR-V (A) was increased at 18:00 and 21:00, the MBR-T (B) and MBR-A (C) were increased at 18:00 compared with control. aP＜0.05.

Figure 4 Brinzolamide-, dorzolamide- and acetazolamide-induced vasodilations Greater relaxation occurred with brinzolamide compared to dorzolamide and acetazolamide (A), representative relaxation curves of acetazolamide (B), dorzolamide (C), and brinzolamide (D).

Figure 5 Effects of L-NAME, indomethacin, and iberiotoxin on brinzolamide-induced relaxation L-NAME (100 μmol/L; A), indomethacin(10 μmol/L; B), or iberiotoxin (0.1 μmol/L; C) were added to vessel segments prior to contraction induced by the high-K solution. There was no significant effect of L-NAME, indomethacin, or iberiotoxin on brinzolamide-induced vasodilatation.

Effect of brinzolamide on histamine-provoked vasoconstrictions in Ca2+-free solutions In Ca2+-free solutions,1 μmol/L histamine induced smooth muscle contractions with or without 1 mmol/L brinzolamide (Figure 6A). We compared the areas under the histamine-induced contraction curves and found that incubation with brinzolamide suppressed histamineprovoked vasoconstrictions in the Ca2+-free solution (n=4,P＜0.05; Figure 6B).

开展工程建设项目审批制度改革，是党中央、国务院推进“放管服”改革、加快转变政府职能的重大决策部署。南京作为全国16个试点地区之一，自试点工作启动以来，全面贯彻习近平新时代中国特色社会主义思想，认真落实党中央、国务院“放管服”改革要求，按照全国工程建设项目审批制度改革试点工作座谈会统一部署，聚焦重点领域和关键环节，对工程建设项目审批制度进行全流程、全覆盖改革，筑造工程建设项目审批制度改革和营商环境优化的“南京模式”“南京品牌”。

DISCUSSION

In the present study, topical application of brinzolamide significantly decreased IOP and increased ODBF in rabbits.These observations raised the question of whether the brinzolamide-augm ented ocular perfusion can be attributed to a primary effect on the ocular vasculature or if it is secondary to IOP reduction. Based on this question, rabbit ocular ciliary arteries were isolated, and brinzolamide was applied to the high-K pre-contracted preparations to measure the development of relaxation ex vivo. The vasodilating capacity of brinzolamide was superior to two other sulfonamide antiglaucoma drugs, dorzolamide and acetazolamide. Furthermore,incubation with brinzolamide suppressed histamine-induced contractions in the Ca2+-free solution.

Figure 6 Effect of brinzolamide on histamine-provoked vasoconstrictions in Ca2+-free solution A: A representative contractile curve in response to histamine in Ca2+-free solution with or without brinzolamide (1 mmol/L); B: We compared the areas under the histamine-induced contraction curves, and found that the responses to histamine with brinzolamide were weaker than without brinzolamide. aP＜0.05.

In vivo, IOP was dec reased bil aterally after bri nzolamide was instilled only in the left eye, though the right eye IOP remained higher than the left. It is unclear how instillation on the left eye can mediate a decrease in contralateral IOP. However, it is possible that after topical instillation in only the left eye, some of the drug was absorbed into the systemic circulation and reached the right eye through that route.

To determine if nitric oxide (NO), prostacyclin, or largeconductance calcium-activated K+ (BKCa) channels were involved in brinzolamide-mediated relaxation, these pathways were blocked with either 100 μmol/L L-NAME, a NO synthase inhibitor[20]; 10 μmol/L indomethacin, a cyclooxygenase inhibitor[21]; or 0.1 μmol/L iberiotoxin, a BKCa channel blocker[22]. These drugs were administrated prior to high-K-induced vasoconstriction. Furthermore, we evaluated the effect of brinzolamide on 1 μmol/L histamine-induced contractions in Ca2+-free solutions. Based on the components of Krebs solution, Ca2+-free solution was prepared by replacing CaCl2 with isotonic equimolar MgCl2 and adding 1 mmol/L ethylene glycol tetraacetic acid (EGTA).

The brinzolamide-induced increase in OBF may be secondary to IOP reduction or it may represent a primary effect on the ocular vasculature. To differentiate between these two possibilities, we studied the effect of brinzolamide on isolated rabbit ciliary arteries. Brinzolamide at clinically relevant concentrations induced relaxation in arteries that were precontracted with high-K solution. These ex vivo results proved the hypothesis that brinzolamide possesses a direct vasodilating effect on the ocular vasculature. Supporting the importance of OBF, Gugleta[24] demonstrated that an improvement of OBF by dorzolamide resulted in preservation of visual fields in glaucoma patients. Thus, there may be an important correlation between the brinzolamide-mediated vasodilation of ocular blood vessels and the preservation of visual function in glaucoma.

We have previously demonstrated that dorzolamide directly induced relaxation of isolated rabbit ciliary arteries[25]. In the present study, we compared the effects of brinzolamide,dorzolamide and acetazolamide ex vivo. Our results showed that acetazolamide could not induce vasodilation, and dorzolamide-induced vascular relaxation was smaller than that induced by brinzolamide. Martinez and Sanchez-Salorio[13]reported that dorzolamide treatment resulted in a 3.8-fold higher peak systolic velocity and a 6.7-fold higher end diastolic velocity in the central retinal artery than did brinzolamide treatment in humans. However, they used the commercially available eye drops to compare the effects between two drugs.In those preparations, the dosing strength of the dorzolamide drops was 2%, which is twice that of the 1% brinzolamide eye drops. In our study, the vasodilating effects of the CAIs were compared at the same concentrations.

The mechanism underlying the direct vasodilating effect of CAIs is controversial. It has been suggested that dorzolamideinduced vasodilation of intraocular porcine ciliary arteries ex vivo depends on NO[26]. However, another study showed that the acetazolamide-induced vasodilation of pial arteriole is sensitive to indomethacin and not on NO release[27].Moreover, Pickkers et al[28] reported that the vasodilator effect of acetazolamide is caused by opening of BKCa channels on vascular smooth muscle cells. In the present study, we found that the concentration-dependent vasodilating effect of brinzolamide is not mediated by NO, prostacyclin, or opening of BKCa channels.

We further investigated the relationship between brinzolamideinduced vasodilation and Ca2+ release from intracellular calcium stores. Histamine provokes smooth muscle contractions using Ca2+ both entry from extracellular space and release from intracellular Ca2+ storages[29]. In Ca2+-free solutions,histamine induces contractions that are mediated by the release of calcium from sarcoplasmic reticulum[30]. We incubated the arterial segments in Ca2+-free solution and found that brinzolamide suppressed histamine-induced contractions. It demonstrated brinzolamide-induced vasodilation is mediated by suppression of Ca2+ release from intracellular calcium stores.

Taken together, our results show that brinzolamide decreased IOP and increased OBF. The direct vasodilatory effect of brizolamide is mediated by suppression of Ca2+ release from intracellular calcium stores. This effect of brinzolamide on OBF may enhance its beneficial action in preserving visual fi eld in glaucoma patients.

ACKNOWLEDGEMENTS

Foundation: Supported by Japan Society for the Promotion of Science (JSPS) Grant-in-Aid for Scientific Research (C)(No.25462750).

Conflicts of Interest: Dong YR, None; Huang SW, None;Cui JZ, None; Yoshitomi T, None.

《枫窗小牍》作者所保留下来的《燕丹子》序言，真可谓小说之隔代知音。既洞烛小说中主要人物的精神气概与诚信人格，又推崇他们的勇士之能，同时指出《燕丹子》不能被衡之以道家与儒家后学的文化观念，它只能是属于那样一个特定时代和特殊文化观念的历史产物。

REFERENCES

1 Silver LH. Dose-response evaluation of the ocular hypotensive effect of brinzolamide ophthalmic suspension (Azopt). Brinzolamide Dose-Response Study Group. Surv Ophthalmol 2000;44(2):S147-S153.

2 DeSantis L. Preclinical overview of brinzolamide. Surv Ophthalmol 2000;44(2):S119-S129.

3 Mincione F, Scozzafava A, Supuran CT. The development of topically acting carbonic anhydrase inhibitors as antiglaucoma agents. Curr Pharm Des 2008;14(7):649-654.

4 Wu R, Yao K, Flammer J, Haefliger IO. Role of anions in nitric oxideinduced short-circuit current increase in isolated porcine ciliary processes.Invest Ophthalmol Vis Sci 2004;45(9):3213-3222.

5 Mastropasqua R, Agnifili L, Mattei PA, Caulo M, Fasanella V,Navarra R, Mastropasqua L, Marchini G. Advanced morphological and functional magnetic resonance techniques in glaucoma. Biomed Res Int 2015;2015:160454.

6 Siesky B, Harris A, Brizendine E, Marques C, Loh J, Mackey J, Overton J, Netland P. Literature review and meta-analysis of topical carbonic anhydrase inhibitors and ocular blood flow. Surv Ophthalmol 2009;54(1):33-46.

7 Schmidl D, Garhofer G, Schmetterer L. The complex interaction between ocular perfusion pressure and ocular blood flow - relevance for glaucoma. Exp Eye Res 2011;93(2):141-155.

8 Siesky B, Harris A, Carr J, Verticchio Vercellin A, Hussain RM, Parekh Hembree P, Wentz S, Isaacs M, Eckert G, Moore NA. Reductions in retrobulbar and retinal capillary blood flow strongly correlate with changes in optic nerve head and retinal morphology over 4 years in openangle glaucoma patients of african descent compared with patients of european descent. J Glaucoma 2016;25(9):750-757.

9 Shiga Y, Kunikata H, Aizawa N, Kiyota N, Maiya Y, Yokoyama Y,Omodaka K, Takahashi H, Yasui T, Kato K, Iwase A, Nakazawa T.Optic nerve head blood flow, as measured by laser speckle fl owgraphy,is significantly reduced in preperimetric glaucoma. Curr Eye Res 2016;41(11):1447-1453.

10 Haustein M, Spoerl E, Boehm AG. The effect of acetazolamide on different ocular vascular beds. Graefes Arch Clin Exp Ophthalmol 2013;251(5):1389-1398.

11 Huber-van der Velden KK, Lux A, Severing K, Klamann MK,Winterhalter S, Remky A. Retrobulbar hemodynamics before and after oculopression with and without dorzolamide. Curr Eye Res 2012;37(8):719-725.

12 Barnes GE, Li B, Dean T, Chandler ML. Increased optic nerve head blood flow after 1 week of twice daily topical brinzolamide treatment in Dutch-belted rabbits. Surv Ophthalmol 2000;44(2):S131-S140.

13 Martinez A, Sanchez-Salorio M. A comparison of the long-term effects of dorzolamide 2% and brinzolamide 1%, each added to timolol 0.5%,on retrobulbar hemodynamics and intraocular pressure in open-angle glaucoma patients. J Ocul Pharmacol Ther 2009;25(3):239-248.

14 Luft N, Wozniak PA, Aschinger GC, Fondi K, Bata AM, Werkmeister RM, Schmidl D, Witkowska KJ, Bolz M, Garhöfer G, Schmetterer L.Ocular Blood Flow Measurements in Healthy White Subjects Using Laser Speckle Flowgraphy. PLoS One 2016;11(12):e0168190.

15 Sugimoto M, Nunome T, Sakamoto R, Kobayashi M, Kondo M. Effect of intravitreal ranibizumab on the ocular circulation of the untreated felloWeye. Graefes Arch Clin Exp Ophthalmol 2017;255(8):1543-1550.

16 Shiba T, Takahashi M, Matsumoto T, Hori Y. Differences in optic nerve head microcirculation between evening and morning in patients with coronary artery disease. Microcirculation 2017;24(7).

17 Dong Y, Ishikawa H, Wu Y, Shimizu K, Goseki T, Yoshitomi T. Effect and mechanism of betaxolol and timolol on vascular relaxation in isolated rabbit ciliary artery. Jpn J Ophthalmol 2006;50(6):504-508.

18 Dong Y, Ishikawa H, Wu Y, Yoshitomi T. Vasodilatory mechanism of levobunolol on vascular smooth muscle cells. Exp Eye Res 2007;84(6):1039-1046.

19 Dong Y, Watabe H, Su G, Ishikawa H, Sato N, Yoshitomi T. Relaxing effect and mechanism of ta fl uprost on isolated rabbit ciliary arteries. Exp Eye Res 2008;87(3):251-256.

20 Yamawaki K, Zamami Y, Kawasaki H, Takatori S. Effects of endogenous nitric oxide on adrenergic nerve-mediated vasoconstriction and calcitonin gene-related peptide-containing nerve-mediated vasodilation in pithed rats. Eur J Pharmacol 2017;802:69-75.

21 Novakovic A, Marinko M, Jankovic G, Stojanovic I, Milojevic P, Nenezic D, Kanjuh V, Yang Q, He GW. Endothelium-dependent vasorelaxant effect of procyanidin B2 on human internal mammary artery.Eur J Pharmacol 2017;807:75-81.

22 Xu YC, Leung SW, Leung GP, Man RY. Kaempferol enhances endothelium-dependent relaxation in the porcine coronary artery through activation of large-conductance Ca(2+) -activated K(+) channels. Br J Pharmacol 2015;172(12):3003-3014.

23 Siesky B, Harris A, Cantor LB, Kagemann L, Weitzman Y, McCranor L, Marques C, Werne A, Stefansson E. A comparative study of the effects of brinzolamide and dorzolamide on retinal oxygen saturation and ocular microcirculation in patients with primary open-angle glaucoma. Br J Ophthalmol 2008;92(4):500-504.

24 Gugleta K. Topical carbonic anhydrase inhibitors and visual function in glaucoma and ocular hypertension. Curr Med Res Opin 2010;26(6):1255-1267.

25 Dong Y, Sawada Y, Cui J, Hayakawa M, Ogino D, Ishikawa M,Yoshitomi T. Dorzolamide-induced relaxation of isolated rabbit ciliary arteries mediated by inhibition of extracellular calcium influx. Jpn J Ophthalmol 2016;60(2):103-110.

26 Kringelholt S, Simonsen U, Bek T. Dorzolamide-induced relaxation of intraocular porcine ciliary arteries in vitro depends on nitric oxide and the vascular endothelium. Curr Eye Res 2012;37(12):1107-1113.

27 Domoki F, Zimmermann A, Tóth-Szuki V, Busija DW, Bari F.Acetazolamide induces indomethacin and ischaemia-sensitive pial arteriolar vasodilation in the piglet. Acta Paediatr 2008;97(3):280-284.28 Pickkers P, Garcha RS, Schachter M, Smits P, Hughes AD. Inhibition of carbonic anhydrase accounts for the direct vascular effects of hydrochlorothiazide. Hypertension 1999;33(4):1043-1048.

29 Sharma A, Nakade UP, Choudhury S, Yadav RS, Garg SK. Extra and intracellular calcium signaling pathway(s) differentially regulate histamine-induced myometrial contractions during early and midpregnancy stages in buffaloes (Bubalus bubalis). Anim Reprod Sci 2017;179:10-19.

30 Morgan SJ, Deshpande DA, Tiegs BC, Misior AM, Yan H, Hershfeld AV, Rich TC, Panettieri RA, An SS, Penn RB. β-Agonist-mediated relaxation of airway smooth muscle is protein kinase A-dependent. J Biol Chem 2014;289(33):23065-23074.