Recent progress on the CRISPR/Cas system in optical biosensors.

Clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated (Cas) protein systems are adaptive immune systems unique to archaea and bacteria, with the characteristics of targeted recognition and gene editing to resist the invasion of foreign nucleic acids. Biosensors combined with the CRISPR/Cas system and optical detection technology have attracted much attention in medical diagnoses, food safety, agricultural progress, and environmental monitoring owing to their good sensitivity, high selectivity, and fast detection efficiency. In this review, we introduce the mechanism of CRISPR/Cas systems and developments in this area, followed by summarizing recent progress on CRISPR/Cas system-based optical biosensors combined with colorimetric, fluorescence, electrochemiluminescence and surface-enhanced Raman scattering optical techniques in various fields. Finally, we discuss the challenges and future perspectives of CRISPR/Cas systems in optical biosensors.

Microcystic adnexal carcinoma of the eyelid and orbit: A case report and review of literature.

Microcystic adnexal carcinoma (MAC), a rare and low-grade malignant skin tumor, is characterized by a high rate of misdiagnosis and a preponderance for local recurrence, but seldom seen nodal or distant metastasis. Although MAC typically occurs almost in the head and neck region, primary eyelid or orbital MAC is very rare. To explore the unique characteristics of the eyelid and orbital MAC, we reviewed the relevant literature. Based on its distinctive anatomical location and the aggressive behavior, eyelid or orbital MAC not only exhibit a high rate of misdiagnosis and local recurrence, but also lead to serious complications such as disfigurement after orbital exenteration, paranasal sinuses or intracranial invasion, even death. Misdiagnosis of MAC commonly result from its rarity and nonspecific clinical and histopathological presentation. To reduce or avoid misdiagnosis, it is important to increase awareness for MAC and obtain a full-thickness biopsy specimen in histopathological analysis. Due to its extensive invasive growth pattern, MAC has a high rate of local recurrence, so complete excision with clear margins and long-term follow-up of patients with MAC are necessary. About those serious complications of the eyelid and orbital MAC, early and accurate diagnosis, complete excision is very important. Moreover, an interprofessional team consisting of ophthalmologist, otolaryngologist, neurologist, dermatologist, pathologist, radiologist is needed to evaluate and treat this disease. In summary, increasing awareness, early and accurate diagnosis, complete excision, long-term follow-up, and a multidisciplinary team is crucial for management of the eyelid and orbital MAC.

Educational degree differences in the association between work stress and depression among Chinese healthcare workers: Job satisfaction and sleep quality as the mediators.

Background: Depressive status of medical personnel worldwide and especially in China is an important public health and social problem. There is a strong relationship between education and depression, but no studies have studied grouping healthcare workers (HCWs) with different educational degree to discuss whether there are differences in the factors that affect depression. This study aims to examine the role of job satisfaction and sleep quality in the relationship between work stress and depression among Chinese HCWs, and teste whether the mediation models are differed by the differences of educational degree. Methods: Patient Health Questionnaire-9 (PHQ-9) scale was used to test depression. Work stress was assessed using the Challenge-blocking stress scale (CBSS). Sleep quality was assessed using the Pittsburgh Sleep Quality Index (PSQI). HCWs' satisfaction with their current work was assessed using the Job Satisfaction Index (JSI). The representative sample of HCWs was chosen using a multi-stage stratified cluster random sampling procedure and 844 HCWs were utilized to the statistical analysis of the study. Results: In the overall sample, sleep quality could mediate the relationship between work stress and depression in healthcare workers (p < 0.001, CMIN/DF = 3.816, GFI = 0.911, AGFI = 0.886, IFI = 0.943, TLI = 0.933, CFI = 0.942, RMSEA = 0.058, SRMR = 0.055, AIC = 1039.144), and the mediating effect accounted for 36.5%. After grouping educational qualifications, the model with sleep quality and job satisfaction as mediating variables reported a better fit in the group with low educational qualifications. The intermediary effect accounted for 50.6 and 4.43%, respectively. The highly educated group only has sleep quality as an intermediary variable in the structural model, and the mediating effect accounted for 75.4% (p < 0.001, CMIN/DF = 2.596, GFI = 0.887, AGFI = 0.857, IFI = 0.937, TLI = 0.926, CFI = 0.937, RMSEA = 0.044, SRMR = 0.056, AIC = 1481.322). Conclusion: In the overall sample, sleep quality could mediate the relationship between work stress and depression in HCWs. Among HCWs with technical secondary school education and below, job satisfaction can mediate the positive relationship between work stress and depression, while this mediating effect is not significant among HCWs with college degree and above.

A review on the oncogenesis of Merkel cell carcinoma: Several subsets arise from different stages of differentiation of stem cell.

Merkel cell carcinoma (MCC), a rare primary cutaneous neuroendocrine neoplasm, is extremely aggressive and has a higher mortality rate than melanoma. Based on Merkel cell polyomavirus (MCPyV) status and morphology, MCCs are often divided into several distinct subsets: pure MCPyV-positive, pure MCPyV-negative, and combined MCC. MCPyV-positive MCC develops by the clonal integration of viral DNA, whereas MCPyV-negative MCC is induced by frequent ultraviolet (UV)-mediated mutations, that are characterized by a high mutational burden, UV signature mutations, and many mutations in TP53 and retinoblastoma suppressor gene (RB1). Combined MCC consists of an intimate mix of MCC and other cutaneous tumor populations, and is usually MCPyV-negative, with rare exceptions. Based on the existing subsets of MCC, it is speculated that there are at least 4 stages in the natural history of stem cell differentiation: primitive pluripotent stem cells, divergent differentiated stem cells, unidirectional stem cells, and Merkel cells (or epidermal/adnexal cells). In the first stage, MCPyV may integrate into the genome of primitive pluripotent stem cells, driving oncogenesis in pure MCPyV-positive MCC. If MCPyV integration does not occur, the stem cells enter the second stage and acquire the ability to undergo multidirectional neuroendocrine and epidermal (or adnexal) differentiation. At this stage, accumulated UV-mediated mutations may drive the development of combined MCC. In the third stage, the stem cells differentiate into unidirectional neuroendocrine stem cells, UV-mediated mutations can induce carcinogenesis in pure MCPyV-negative MCC. Therefore, it has been speculated that several subsets of MCCs arise from different stages of differentiation of common stem cells.

Removal of iatrogenic orbital foreign bodies with a 3D model.

Facial foreign body (FB) is common after trauma, but iatrogenic orbital FB is a rare and unexpected complication of facial FB removal surgery. We present the case of a 43-year-old man with a glass FB in his nose. During the operation, this FB broke into two pieces, and the larger one pierced into the left orbit, close to the eyeball. A three-dimensional (3D) model was made that accurately recreated the shape and position of the FB in the orbit, according to which the FB was removed. 3D-printing technology is a great tool when dealing with complex facial FB.

The Mediating Effect of Perceived Social Support on Mental Health and Life Satisfaction among Residents: A Cross-Sectional Analysis of 8500 Subjects in Taian City, China.

Several studies have explored the relationship between mental health and life satisfaction. However, few studies have clarified the mechanisms underlying the relationship between mental health and life satisfaction among a large sample of the whole population. The aim of this study was to explore the mediating role of perceived social support between mental health and life satisfaction among the residents in Taian City, China. A total of 8500 residents were included in the analysis. A descriptive analysis was conducted to describe the sample characteristics. Pearson correlation was employed to explore the correlation between mental health and life satisfaction. The mediating role of perceived social support was analyzed using SPSS26.0. This study found that the residents' average score of life satisfaction was 24.60 ± 4.12. Mental health was significantly correlated with perceived social support and life satisfaction. After adjusting for controlling variables, perceived social support played a partially mediating effect on mental health and life satisfaction, accounting for 21.04% of the total effect. However, data are cross-sectional, and causal conclusions cannot be drawn. Attention should be paid to the residents' mental health and intervention should be considered for residents with mental disorders to improve the residents' life satisfaction.

MiR-421 Aggravates Neurotoxicity and Promotes Cell Death in Parkinson's Disease Models by Directly Targeting MEF2D.

Parkinson's disease (PD) is a severe neurodegenerative disease characterized by selective loss of dopaminergic neurons, which reduces quality of life of patients and poses a heavy burden to the society. The pathological mechanism of PD remains unclear, and increasing efforts are aimed to solve this problem. MiRNAs are a kind of small noncoding RNA regulating target gene expression. Previous studies have shown that dysregulation of miRNAs is involved in the development of PD. In the present study, we determined that miR-421 and MEF2D are increased and decreased, respectively, in a cellular model of PD. The data on the mechanism of action indicate that miR-421 directly binds to MEF2D mRNA and negatively regulates MEF2D expression. An increase in miR-421 disrupted the Bcl2/Bax system. Functional assays indicated that enhanced miR-421 promotes cell death by negative modulation of MEF2D expression. Inhibition of miR-421 or restoration of MEF2D protected neurons from neurotoxicity in cellular and animal models of PD. Our study is the first to demonstrate that miR-421 is decreased in PD models and to determine a novel putative mechanism of PD pathogenesis.

Remodeled structure and reduced contractile responsiveness of ocular ciliary artery in spontaneously hypertensive rats.

AIM: To investigate the alterations in both structure and contractile responsiveness of ocular ciliary artery (OCA) in spontaneously hypertensive rat (SHR). METHODS: In this experiment, 20-week-old male SHR and Wistar Kyoto rat (WKY) were studied. The heart rate (HR), the blood pressure (BP; the systolic BP and the diastolic BP) of rats with an electronic sphygmomanometer were measured. Vascular morphometry and isometric tension measurement were used to investigate the alterations in structure and contractility of OCA. RESULTS: A general narrowing of OCAs was observed in SHR compared to the control WYK. In SHR, the media of OCAs were thicker, the luminal diameters were smaller, and the media-to-lumen ratios were higher when compared with WKY (P<0.05). The contractions of OCAs evoked by norepinephrine were smaller in SHR compared to control (P<0.05). Then, OCAs were pretreated with iberiotoxin, L-NAME, or indomethacin 30min before norepinephrine-induced contraction. Iberiotoxin (0.1 µmol/L) has not changed the norepinephrine-induced contractions in OCAs from both groups. However, L-NAME (100 µmol/L) increased the vasoconstrictions, the increased extents were similar in SHR and WKY (P>0.05). Indomethacin (10 µmol/L) decreased the contractions induced by norepinephrine in OCAs from WKY (P<0.05), but did not change those contractions in vessels from SHR (P>0.05). CONCLUSION: Our results demonstrate that the structure and function of OCAs are altered in hypertension. OCAs from SHR are remodeled with decreased lumen diameter and increased media-to-lumen ratio. Moreover, the contractile responsiveness of OCAs from SHR is diminished due to the disruption of vasoconstrictive effect of prostaglandins.

Effects of brinzolamide on rabbit ocular blood flow in vivo and ex vivo.

AIM: To investigate if significant improvement of optic disc blood flow (ODBF) occurs after instillation of brinzolamide onto rabbit eyes. METHODS: Testing of bilateral intraocular pressure (IOP) and left ODBF in 10 male rabbits took place every 3h over a 24h period. Brinzolamide (1% ophthalmic solution, two drops at 9:00 and 21:00) was administered to the left eye. ODBF, assessed using laser speckle flowgraphy, was determined as the mean blur rate (MBR). Furthermore, the effect of brinzolamide on isolated rabbit ciliary arteries using isometric tension recording system was performed. RESULTS: After brinzolamide instillation, IOP was significantly decreased in the left eye. MBR-vessel was greater at 18:00 and 21:00 (P<0.05) than in the controls. MBR-tissue and MBR-average were greater at 18:00 (P<0.05) than in the controls. For isolated arteries pre-contracted with a high-K solution, brinzolamide induced concentration-dependent relaxation, reaching 46.1%±9% (n=21) at 1 mmol/L. In Ca2+-free solutions, incubation with brinzolamide suppressed 1 µmol/L histamine-induced contractions (P<0.05). CONCLUSION: Brinzolamide decreases IOP and increases ocular blood flow. The direct vasodilatory effect of brizolamide is mediated by suppression of Ca2+ release from intracellular calcium stores.

Dacryocystitis due to sporothrix inoculated vis an unusual mode: Case report.

RATIONALE: Sporotrichosis is the most common subcutaneous mycosis. It is caused by the dimorphic fungus Sporothrix schenckii. Ocular sporotrichosis is uncommon and has been rarely reported. PATIENT CONCERNS: We describe a 34-year-old female who presented with a nodule increasing in size near the medial angle of the left eye. Originally, she was misdiagnosed with a dacryocyst space-occupying lesion, and the lesion was removed by surgery. DIAGNOSES: Findings of fungal structures in the histopathological examination contributed to the diagnosis of Sporothrix dacryocystitis. Further culture of conjunctival secretions and contact lens storage solution was positive for Sporothrix. INTERVENTIONS: She was treated with oral itraconazole, 200 mg by mouth twice daily. OUTCOMES: After 3 months of treatment with oral itraconazole, culture of the conjunctival secretions was negative. LESSONS: It is of paramount importance to clinically suspect mycosis, even in unusual locations or in the absence of the typical epidemiological history.

LncRNA SNHG1 regulates cerebrovascular pathologies as a competing endogenous RNA through HIF-1α/VEGF signaling in ischemic stroke.

Studies have shown that long noncoding ribonucleic acids (lncRNAs) play critical roles in multiple biologic processes. However, the Small Nucleolar RNA Host Gene 1 (SNHG1) function and underlying molecular mechanisms in ischemic stroke have not yet been reported. In the present study, we found that SNHG1 expression was remarkably increased both in isolated cerebral micro-vessels of a middle cerebral artery occlusion (MCAO) mice model, and in oxygen-glucose deprivation (OGD)-cultured mice brain micro-vascular endothelial cells (BMECs), meanwhile, the SNHG1 level was negatively correlated with miR-18a in MCAO mice. Mechanistically, SNHG1 inhibition presents larger brain infarct size and worsens neurological scores in MCAO mice. Consistent with the in vivo findings, SNHG1 inhibition also significantly increased caspase-3 activity and cell apoptosis in OGD-cultured BMECs. Furthermore, we found that SNHG1 functions as a competing endogenous RNA (ceRNA) for miR-18a, thereby regulating the de-repression of its endogenous target HIF-1α and promoting BMEC survival through HIF-1α/VEGF signaling. This study found a neuroprotective effect of SNHG1 mediated by HIF-1α/VEGF signaling through acting as a ceRNA for miR-18a. These findings reveal a novel function of SNHG1, which contributes to an extensive understanding of ischemic stroke and provides novel therapeutic options for this disease.

Microstructure, friction and corrosion resistance properties of a Ni-Co-Al2O3 composite coating.

Ni-Co-Al2O3 composite coatings were prepared by pulsed electrodeposition and electrophoresis-electrodeposition on aluminum alloy. The content of Al2O3 particles of the Ni-Co-Al2O3 composite coating prepared by electrophoresis-electrodeposition was significantly higher than the composite coating prepared by pulsed electrodeposition. The composite coating prepared by electrophoresis-electrodeposition exhibited a better anti-wear performance than that prepared by pulsed electrodeposition. The morphology, composition and microstructure of the composite coatings were determined by means of X-ray diffractometer (XRD) and scanning electron microscopy (SEM). The hardness and friction properties of the samples were tested on the microhardness tester and the friction and wear loss tester respectively.

NPPB modulates apoptosis, proliferation, migration and extracellular matrix synthesis of conjunctival fibroblasts by inhibiting PI3K/AKT signaling.

When treating glaucoma, excessive scar tissue reactions reduce the postoperative survival rate of the filtering bleb. Accumulating evidence has demonstrated that the proliferation, migration and extracellular matrix (ECM) synthesis of fibroblasts are important molecular mechanisms underlying scar formation. Recent evidence has demonstrated that chloride channels play an important role in controlling cell proliferation, apoptosis, migration and the cell cycle process in several cell types, but the effects of chloride channels on conjunctival fibroblasts have not be studied. The aim of the present study was to investigate the effects of the chloride channel blocker 5-nitro-2-(3-phenylpropylamino) benzoic acid (NPPB) on cell proliferation, apoptosis, migration, cell cycle progression and ECM synthesis in human conjunctival fibroblasts (HConFs), and to further investigate the mechanism of resistance to scar formation following glaucoma filtration surgery. HConFs were exposed to NPPB or lubiprostone. Cell proliferation and viability was evaluated using the Cell Counting Kit-8. Cell migration was measured using Transwell migration and scratch­wound assays. Flow cytometry was used to study apoptosis and cell cycle progression. Quantitative polymerase chain reaction and western blot analyses were performed to determine mRNA and protein expression levels, respectively. Following NPPB treatment, HConFs exhibited reduced proliferation and migration, along with increased apoptosis. NPPB also inhibited cell cycle progression by arresting cells in the G0̸G1 phase and reducing collagen I and fibronectin expression, as well as the phosphorylation of phosphoinositide 3-kinase (PI3K) and protein kinase B (AKT). However, lubiprostone treatment exerted the opposite effects on HConFs. Therefore, NPPB treatment inhibited proliferation, migration, cell cycle progression and synthesis of the ECM, while promoting apoptosis in HConFs, by inhibiting the PI3K̸AKT signaling pathway.

Long noncoding RNAs coordinate functions between mitochondria and the nucleus.

In animal cells, mitochondria are the primary powerhouses and metabolic factories. They also contain genomes and can produce mitochondrial-specific nucleic acids and proteins. To maintain homeostasis of the entire cell, an intense cross-talk between mitochondria and the nucleus, mediated by encoded noncoding RNAs (ncRNAs), as well as proteins, is required. Long ncRNAs (lncRNAs) contain characteristic structures, and they are involved in the regulation of almost every stage of gene expression, as well as being implicated in a variety of disease states, such as cancer. In the coordinated signaling system, several lncRNAs, transcribed in the nucleus but residing in mitochondria, play a key role in regulating mitochondrial functions or dynamics. For example, RMRP, a component of the mitochondrial RNase MRP, is important for mitochondrial DNA replication and RNA processing, and the steroid receptor RNA activator, SRA, is a key modulator of hormone signaling and is present in both the nucleus and mitochondria. Some RNA-binding proteins maybe play a role in the lncRNAs transport system, such as HuR, GRSF1, SHARP, SLIRP, PPR, and PNPASE. Furthermore, a series of nuclear DNA-encoded lncRNAs were implicated in mitochondria-mediated apoptosis, mitochondrial bioenergetics and biosynthesis, and glutamine metabolism. The mitochondrial genome can also encode a set of lncRNAs, and they are divided into three categories: (1) lncND5, lncND6, and lncCyt b RNA; (2) chimeric mitochondrial DNA-encoded lncRNAs; and (3) putative mitochondrial DNA-encoded lncRNAs. It has been reported that the mitochondrial DNA-encoded lncRNAs appear to operate in the nucleus. The molecular mechanisms underlying trafficking of the mitochondrial DNA-encoded lncRNAs to the nucleus in mammals are only now beginning to emerge. In conclusion, both nuclear- and mitochondrial DNA-encoded lncRNAs mediate an intense intercompartmental cross-talk, which opens a rich field for investigation of the mechanism underlying the intercompartmental coordination and the maintenance of whole cell homeostasis.

Hypoperfusion retinopathy and elevated intraocular pressure in a 17-year-old.

We describe a case of bilateral hypoperfusion retinopathy (HR) in a 17-year-old man with intraocular pressure (IOP) of >40 mm Hg. After 6 months of antiglaucoma drug therapy, the patient's bilateral IOP remained over 35 mm Hg, and features of HR were found bilaterally in the absence of carotid artery obstruction. On restoration of IOP to normal by means of trabeculectomy, the bilateral signs of HR regressed. We speculate that the elevated IOP contributed to HR in this patient.

Antimicrobial and antioxidant activities of Flammulina velutipes polysaccharides and polysaccharide-iron(III) complex [corrected].

FVP is polysacchrides obtained from Flammulina velutipes. A polysacchride named FVP2 was isolated from FVP by DEAE cellulose-52 chromatography and Sephadex G-100 size-exclusion chromatography. FVP-Fe and FVP2-Fe were synthesized by neutralization of FeCl3 carbohydrate solution. The antibacterial and antifungal activities of FVP, FVP2, FVP-Fe, FVP2-Fe were investigated and their antioxidant effects on hydroxyl, 2,2-diphenyl-1-picrylhydrazyl (DPPH), superoxide anion, 2,2'-azobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radicals, reducing power, inhibition of malondialdehyde (MDA) were assessed in vitro. The results suggested that FVP-Fe and FVP2-Fe significantly suppressed the growth of bacteria Staphylococcus aureus, Escherichia coli, and Bacillus subtilis, and have relatively strong antioxidant activity to scavenge superoxide anion radical. In addition, FVP exhibited strong antioxidant activity to eliminate hydroxyl, DPPH, ABTS radicals, had high reducing power and inhibited the MDA production of health mice liver homogenate induced by auto-oxidation and Fe2+-H2O2 system.

Anti-apoptotic and angiogenic effects of intelectin-1 in rat cerebral ischemia.

Ischemic stroke is an acute life-threatening disease, which causes neurological dysfunction. The formation of new blood vessels around the infarct is vital to the restoration of perfusion and healing of brain tissue. Studies have shown that intelectin-1 (omentin) promotes endothelial cell function and angiogenesis in response to ischemia and inhibits apoptosis in rats with unilateral hind limb surgery. In the present study, we investigated the neuroprotective role of intelectin-1 following focal cerebral ischemia. We specifically assessed the effect of increased expression of intelectin-1 in promoting angiogenesis and reducing apoptosis. The treatment was administered using a lentiviral vector, 7 days prior to surgery. The surgery was performed using the established middle cerebral artery occlusion (MCAO) model in rats, and the outcome was evaluated 7 days after injury. Our results demonstrated a significant reduction in brain infarction volume following LV-intelectin-1 treatment. Additionally, CD34 and capillary density were increased in the cerebral ischemic penumbra. Real-time PCR and Western blot revealed an increased expression of intelectin-1, and phosphorylation of eNOS and AKT with enhanced expression of bcl-2 in brain tissues. These data suggest that the successful delivery of LV-intelectin-1 ameliorated ischemic brain injury. It promoted endothelial cell function and revasc ularization, and inhibited apoptosis in response to ischemia by stimulating the Akt-eNOS signaling pathway.

The CLC-2 Chloride Channel Modulates ECM Synthesis, Differentiation, and Migration of Human Conjunctival Fibroblasts via the PI3K/Akt Signaling Pathway.

Recent evidence suggests that chloride channels are critical for cell proliferation, migration, and differentiation. We examined the effects of transforming growth factor (TGF)-ß1 on chloride channel expression and associations with human conjunctival fibroblast (HConF) biology. To investigate the potential role of chloride channel (CLC)-2 in migration, transition to myofibroblasts and extracellular matrix (ECM) synthesis of HconF, a small interfering RNA (siRNA) approach was applied. TGF-ß1-induced migration and transition of fibroblasts to myofibroblasts characterized by α-smooth muscle actin (α-SMA) expression, supported by increased endogenous expression of CLC-2 protein and mRNA transcripts. ECM (collagen I and fibronectin) synthesis in HConF was enhanced by TGF-ß1. CLC-2 siRNA treatment reduced TGF-ß1-induced cell migration, transition of fibroblasts to myofibroblasts, and ECM synthesis of HConF. CLC-2 siRNA treatment in the presence of TGF-ß1 inhibited phosphorylation of PI3K and Akt in HConF. These findings demonstrate that CLC-2 chloride channels are important for TGF-ß1-induced migration, differentiation, and ECM synthesis via PI3K/Akt signaling in HConF.

Dorzolamide-induced relaxation of isolated rabbit ciliary arteries mediated by inhibition of extracellular calcium influx.

PURPOSE: The carbonic anhydrase inhibitor dorzolamide can increase optic nerve blood flow. The aim of the study reported here was to investigate the effect of dorzolamide on isolated rabbit ciliary arteries that supply the optic nerve. METHODS: Changes in ciliary artery isometric tension and intracellular Ca(2+) concentration ([Ca(2+)]i) were recorded to elucidate the underlying pharmacologic mechanisms by which dorzolamide regulates blood flow to the optic nerve. RESULTS: Dorzolamide induced concentration-dependent relaxation of rabbit ciliary arteries that had been precontracted by exposure to a high potassium (high-K) solution. Neither pretreatment with 10 µM KB-R 7943, an Na(+)/Ca(2+) exchanger inhibitor, nor alkalinization of the high-K solution had an effect on the dorzolamide-induced relaxation. Pretreatment with 100 µM N(G)-nitro-L-arginine methylester, a nitric oxide synthase inhibitor (n = 10), 10 µM indomethacin, a prostacyclin inhibitor (n = 9), or 0.1 µM iberiotoxin, an inhibitor of endothelium-derived hyperpolarizing factor (n = 7), did not change the concentration-dependent relaxation induced by dorzolamide. Incubation with 3 mM dorzolamide in a Ca(2+)-free solution did not change the transient contractions of the rabbit ciliary arteries induced by 1 µM histamine (n = 9). However, 3 mM dorzolamide significantly suppressed the increase in [Ca(2+)]i induced by the reintroduction of Ca(2+) to a calcium-free extracellular medium (P < 0.05). Furthermore, 3 mM dorzolamide significantly suppressed the [Ca(2+)]i increase induced by the high-K solution (P < 0.05). CONCLUSIONS: Taken together, our results reveal a novel role for dorzolamide in relaxing the ciliary arteries. Our data support the hypothesis that the vasodilatory action of dorzolamide is mediated by inhibition of Ca(2+) entry through voltage-dependent Ca(2+) channels.