A sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated to quantify atropine in ten rabbit ocular tissues enabling systematic characterization of the ocular pharmacokinetic profile of 0.01% atropine sulfate eye drops after a single topical administration. The method demonstrated excellent linearity (coefficient of determination, R2 ≥ 0.9908) across all matrices, with lower limits of quantification (LLOQ) of 0.05 ng/mL for most tissues and 0.10 ng/mL for retina and lens; intra- and inter-day accuracy, precision, matrix effects, extraction recoveries, and stability all met the acceptance criteria. Following a single bilateral topical dose (50 μL/eye) in New Zealand White rabbits, atropine distributed rapidly into all 12 ocular compartments (the sclera further divided into three anatomical regions) with marked heterogeneity-the highest exposures were found in conjunctiva and cornea, a distinct anterior-to-posterior concentration gradient was observed in the sclera, sustained retention was noted in the retina (mean residence time from zero to the last measurable time point, MRT0-t 3.30 h), while aqueous and vitreous humor eliminated rapidly (elimination half-life, t₁/₂ < 0.7 h), and all tissues except aqueous humor followed a two-compartment model. This validated method and the comprehensive pharmacokinetic data reveal that topically applied 0.01% atropine achieves sustained exposure in key myopia-regulating tissues (retina, choroid, posterior sclera) with low exposure in side-effect target tissues (iris, ciliary body, lens).

Fungal keratitis (FK) is a persistent and vision-threatening disease. The potential of vitamin C (VC) in tissue protection is well-recognized; however, its specific role in FK and its interaction with antifungal drugs are not well defined. In this study, we investigated the role of VC in FK in vitro and in vivo. It revealed that VC substantially decreases clinical scores and corneal perforation rates in a murine model of FK. VC promoted a concentration-dependent increase in neutrophil infiltration via mast cells in vitro and confirmed this effect in in vivo experiments. Moreover, VC enhanced fungicidal activity by boosting infiltrated neutrophils, without influencing reactive oxygen species (ROS) production or neutrophil extracellular traps (NETs) formation. VC also mitigated the cytokine storm within bone marrow-derived macrophages and increased neutrophil apoptosis, thereby facilitating the efficient clearance of senescent neutrophils. The combination of VC with amphotericin B (AmB) demonstrated additive antifungal effects, reducing fungal load and corneal perforation. These results indicate that VC is pivotal in defending against fungal corneal infections by promoting neutrophil chemotaxis through mast cells (MCs) and modulating the inflammatory response, without suppressing neutrophils' antifungal capability. The combination of VC with AmB may present a new therapeutic avenue for FK.

BACKGROUND:Progastrin-releasing peptide (proGRP) is a potential high-specificity biomarker for Ewing sarcoma and has important clinical value in early diagnosis and risk stratification. However, the existing detection methods such as enzyme-linked immunosorbent assay (ELISA), chemiluminescence immunoassay (CLIA), and cytometric bead array (CBA) often face the problems of insufficient sensitivity and limited reliability in complex biological matrices, lead to the difficulty of accurate quantification. Therefore, it is of great significance to develop new sensing strategies with higher detection performance for promoting clinical accurate diagnosis. RESULTS:This study developed a novel biosensing system based on fluorescence enhancement with Förster resonance energy transfer (FRET) for highly sensitive detection of proGRP. The signal amplification unit of this system is composed of DNA1-modified upconversion nanoparticles (UCNPs) as FRET donor and DNA2-modified magnetic nanozyme complexes (Fe3O4Ns) as acceptor. Upon competitive binding with proGRP, the dissociated DNA1-UCNPs pair with DNA2-Fe3O4Ns. This pairing reduces the donor-acceptor distance, thus activating the FRET process and prompting Fe3O4Ns to generate substantial hydroxyl radicals (·OH), which subsequently oxidizes specific substrates to transduce the detection signal. The detection limit of the method was 0.14 pg/mL in PBS and 0.76 pg/mL in serum. The method demonstrated high specificity, excellent reproducibility, and robust anti-interference performance, rendering it robust for the analysis of complex serum matrices. SIGNIFICANCE:This work presents an innovative sensing platform that combines near-infrared light-excited luminescence with nanozyme-mediated signal amplification for the detection of proGRP. The method surpasses conventional assays in sensitivity and operational stability, showing great promise for clinical translation. Its robust performance in serum underscores potential applications in early cancer diagnostics and biomarker-based disease management, providing a valuable tool for disease diagnosis.

Purpose: To assess the potential of neutrophil-to-lymphocyte ratio (NLR) and platelet-to-lymphocyte ratio (PLR) as novel diagnostic and prognostic biomarkers in fungal keratitis (FK). Methods: This study was carried out retrospectively in 77 FK patients and 77 matched cataract controls from Henan Eye Hospital. Peripheral venous blood samples were collected via venipuncture and analyzed using complete blood count for routine clinical evaluation. FK patients were classified into three subgroups: Fusarium, Aspergillus, and Candida groups. Inflammation severity was quantified using standardized clinical scoring. The treatment modalities were used to divide the FK patients into enucleation and nonenucleation groups. Results: NLR and PLR were significantly elevated in FK versus controls (p < 0.001). NLR correlated strongly with inflammation scores (r = 0.535, p < 0.0001), exceeding PLR's moderate correlation (r = 0.311, p=0.0059). FK patients in the enucleation group had significantly higher NLR (p=0.012) and PLR (p=0.021) values than those in the nonenucleation group. There were no significant biomarker differences across fungal species (p > 0.05). Conclusion: Elevated NLR and PLR values during routine laboratory testing might serve as supplementary indicators for early suspicion of FK and monitoring inflammatory progression, particularly in resource-limited settings where specialized ophthalmic diagnostics are unavailable.

This systematic review and bibliometric analysis investigated the keywords appearing most in the pregnancy-related eye disease field to elucidate the current state and trends of pregnancy-related eye disease research. A systematic literature analysis of pregnancy-related eye disease was performed using the Web of Science Core Collection (WOS) databases. We used the keywords “ocular” OR “eye*” and “pregnancy” OR “pregnant” OR “gestation” to search for articles published from 1999 to 2022. Study data were analyzed and visualized using VOSviewer and CiteSpace. We analyzed 929 articles published from 1999 to 2022. From 1999 to 2012, the article number increased slowly, with a marked acceleration in publication frequency after 2013, original papers accounted for 780 (84

Background: This bibliometric analysis explored the knowledge structure of and research trends in the relationship between light and myopia. Methods: Relevant literature published from 1981 to 2024 was collected from the Web of Science Core Collection database. Visual maps were generated using CiteSpace and VOSviewer. We analyzed the included studies in terms of the annual publication count, countries, institutional affiliations, prolific authors, source journals, top 10 most cited articles, keyword co-occurrence, and cocitations. Results: A total of 525 papers examining the relationship between light and myopia published between 1981 and 2024 were collected. The United States ranked first in terms of the number of publications and actively engaged in international cooperation with other countries. The New England College of Optometry, which is located in the United States, was the most active institution and ranked first in terms of the number of publications. Schaeffel Frank was the most prolific author. The most active journal in the field was Investigative Ophthalmology & Visual Science. The most frequently cited paper in the included studies was written by Saw, SM and was published in 2002. The most common keywords in basic research included “refractive error,” “longitudinal chromatic aberration,” and “compensation.” The most common keywords in clinical research mainly included “light exposure,” “school,” and “outdoor activity.” The current research hotspots in this field are “progression,” “refractive development,” and “light exposure.” The cocitation analysis generated 17 clusters. Conclusion: This study is the first to use bibliometric methods to analyze existing research on the relationship between light and myopia. In recent years, the intensity and wavelength of light have become research hotspots in the field. Further research on light of different intensities and wavelengths may provide new perspectives in the future for designing more effective treatments and interventions to reduce the incidence of myopia.

Objective:To investigate the mechanism of tissue damage caused by neutrophil matrix metalloproteinase-8 (MMP-8) in Fusarium keratitis. Methods:A total of 108 male C57BL/6J SPF grade mice, 6-8 weeks old, were selected to establish a model of Fusarium keratitis (FK) in the right eyes.Corneal inflammation in mice was observed and scored under a slit lamp microscope.Based on the corneal inflammation scores, the modeling eyes were divided into 0, 12, 24, 48, and 72-hour groups post-modeling.At the corresponding time points, mice were euthanized, and corneal tissues were collected.The expressions of MMP-8, adenylate-activated protein kinase (AMPKα) and its serine 172-site phosphorylated form (p-AMPKα) proteins in corneal tissues were detected by Western blot.The neutrophil count in mice corneal tissues at each time point was determined using hematoxylin and eosin staining.The co-localization of neutrophils and MMP-8 protein in the cornea was observed by immunofluorescence staining.In the in vitro corneal collagen degradation experiment, corneal tissues were divided into MMP-8 group, buffer group, and normal saline group, which were treated with 100 μl of activated recombinant MMP-8, detection buffer, and normal saline, respectively.Hydroxyproline content in corneal tissues was determined using a hydroxyproline assay kit, and the mass fractions of hydroxyproline were compared among the groups.Peripheral blood neutrophils were isolated from human blood samples, and Fusarium spores were collected for experiments.Human neutrophils were divided into four groups, negative control group (cultured neutrophils), co-culture group (neutrophils co-cultured with spores), AICAR-treated group (neutrophils co-cultured with spores and treated with p-AMPK protein kinase activator AICAR), and compound C-treated group (neutrophils co-cultured with spores and treated with the inhibitor compound C).The MMP-8 protein expression levels in each group of human neutrophils were assessed via immunofluorescence staining.The use and care of animals complied with the ARVO statement and Regulations for the Administration of Affairs Concerning Experimental Animals.The animal experiment protocol was approved by the Animal Ethics Committee of Henan Eye Hospital (No.HNEECA-2017-04-02).One healthy adult volunteer was selected and 10 ml of peripheral venous blood was collected.The clinical study protocol was approved by the Clinical Ethics Committee of Henan Eye Hospital (No.HNEECKY-2019[16]). Results:At 24 hours post-modeling, corneal opacification was observed in the modeling eyes, and corneal perforation occurred in 72-hour post-modeling group.The corneal inflammation scores in 24, 48, and 72-hour post-modeling groups were all higher than those in 12-hour post-modeling group, and the differences were statistically significant (all at P<0.001).The relative expression levels of MMP-8 protein in the cornea were higher in 12, 24, and 48-hour post-modeling groups compared to 0-hour group, with statistically significant differences (all at P<0.001).There was a moderate positive correlation between the relative expression level of MMP-8 protein in the cornea and the inflammation scores of the modeling eye ( rs=0.50, P<0.05).In the cornea, the p-AMPKα (Thr 172)/AMPKα ratio was higher in 24, 48, and 72-hour post-modeling groups than in 0-hour group, and the differences were statistically significant (all at P<0.05).The p-AMPKα(Thr 172)/AMPKα ratio in the cornea was moderately positively correlated with the relative expression level of MMP-8 protein ( r=0.54, P<0.01).The number of neutrophils in the cornea was significantly higher in 24, 48, and 72-hour post-modeling groups than in 0-hour group, with statistically significant differences (all at P<0.001).The number of neutrophils in the cornea was strongly positively correlated with the inflammation score ( rs=0.77, P<0.001), and was moderately positively correlated with the relative expression level of MMP-8 protein ( r=0.56, P<0.05).MMP-8 protein expression in the cornea of the modeling eyes showed a high degree of co-localization with neutrophils.The hydroxyproline content in the cornea was (0.52±0.02)μg/mg, (0.51±0.03)μg/mg, and (0.27±0.02)μg/mg in buffer group, normal saline group and MMP-8 group, respectively, with a significant overall difference among them ( F=156.63, P<0.01).The corneal hydroxyproline content was lower in MMP-8 group compared to buffer and normal saline groups, and the differences were statistically significant (all at P<0.05).In the experiment involving the infection of cultured Fusarium spores with human neutrophils, the fluorescence intensity of MMP-8 expression was significantly higher in AICAR-treated group than in negative control group and compound C-treated group, with statistically significant differences (all at P<0.05). Conclusions:The MMP-8 secreted by neutrophils in mice with fungal keratitis can degrade corneal stromal collagen fibers, leading to corneal opacification or perforation.The variations in MMP-8 protein expression levels in human neutrophils may be associated with AMPK activation.

患儿，男，10个月余，汉族，因筛查眼部病变于2021年12月6日就诊于河南省立眼科医院。患儿为第3胎，顺产，妊娠35周+4天出生，出生体质量2 150 g，孕妇自觉孕期胎动少。既往有新生儿高胆红素血症史，新生儿期喂养困难，少哭闹。患儿父母非近亲婚配，均体健，患儿1胞姊1胞兄均身体健康；家族中无类似疾病患者，否认家族其他遗传病史。患儿智力发育迟缓、眼睛外观无异常、头发浓密、眼距宽、眉毛浓密下斜、睫毛长(图1)，高腭弓，皮肤白皙，腰、臀部见片状蒙古斑，多线掌纹，掌骨和跖骨短，指趾甲发育不良，关节松弛，四肢肌张力低。眼部检查：角膜映光法检查反光点居中；裂隙灯显微镜检查结膜、角膜无异常；双眼晶状体下半部皮质点状混浊；自然瞳孔下电脑验光显示右眼+1.50 DS/－3.00 DC×9°；左眼+2.00 DS/－3.25 DC×178°；检影验光显示右眼+2.00 DS/－2.50 DC×5°，左眼+2.00 DS/－2.50 DC×180°；眼底照相显示双眼视盘圆，边界清，血管走形和发育可，中心凹反光点存在，视网膜色素不均匀，提示视网膜色素变性(图2)；光学相干断层扫描(optical coherence tomography，OCT)检查显示双眼黄斑及后极部外核层变薄，光感受器层消失(中心凹部分区域除外)，视网膜色素上皮(retinal pigment epithelium，RPE)层粗糙变薄，脉络膜变薄，左眼中心凹神经上皮层可见囊样表现(图3)。对患儿及其父母行全基因外显子检测显示患儿存在 VPS13B基因的1个纯合变异：c.6940+1(IVS38)G>T，其父母为表型正常的杂合变异携带者；线粒体DNA高敏感性测序分析为阴性，排除线粒体病；氨基酸、肉碱谱检测以及尿有机酸谱检测排除遗传代谢病。结合患儿的临床特征、基因检测、线粒体DNA、氨基酸、肉碱谱检测以及尿有机酸谱检测，诊断为Cohen综合征。

Objective:To investigate the expression of adenosine 5'-monophosphate-activated protein kinase (AMPK) phosphorylation in corneal epithelial cells and the effects of fungus on AMPK phosphorylation and interleukin-6 (IL-6) production in corneal epithelial cells.Methods:The human immortalized corneal epithelial cell line was selected.The safe concentration range of AMPK agonist 5-aminoimidazole-4-carboxamide 1-β-D-ribofuranoside (AICAR) (100, 300, 500, 1 000 μmol/L) and inhibitor Compound C (10.0, 12.5, 15.0, 17.5, 20.0 μmol/L) on corneal epithelial cells was screened by multi-function real-time unlabeled cell analyzer.Corneal epithelial cells without any treatment were used as the normal control group, and those co-cultured with spores were used as the spore control group.Corneal epithelial cells co-cultured with spores were treated with AICAR and Compound C for 4 hours in the AICAR group and Compound C group, respectively.The expression of phosphorylated AMPK (p-AMPK) and AMPK in corneal epithelial cells was detected by Western blot, and the concentration of IL-6 in the culture supernatant was determined by enzyme-linked immunosorbent assay (ELISA).Results:After treatment with different concentrations of AICAR for different periods, there was no statistical significance in the cell index of corneal epithelial cells (all at P>0.05). The cell index of corneal epithelial cells was increased with 10.0 μmol/L and 12.5 μmol/L Compound C treatment compared with that of the normal control group.The expression levels of p-AMPK were 0.67±0.15, 2.57±0.12, 3.67±0.58 and 1.50±0.50, respectively, in the normal control group, spore control group, AICAR group and Compound C group, showing a statistically significant difference among them ( F=32.820, P<0.001). The expression level of p-AMPK was significantly higher in the spore control group compared with the normal control group ( P<0.001). The expression level of p-AMPK in the AICAR group was higher than that in the spore control group, and the expression level of p-AMPK in the Compound C group was lower than that in the spore control group, and the differences were statistically significant (both at P=0.010). There was no significant difference in the relative expression level of AMPK among the four groups ( F=0.120, P=0.950). The expression levels of IL-6 concentration in the normal control group, spore control group, AICAR group and Compound C group were (107.81±17.15), (156.32±9.94), (167.96±14.16) and (127.42±19.75)pg/ml, respectively, showing a statistically significant difference among them ( F=15.210, P<0.001). The IL-6 concentration of the spore control group was higher than that of the normal control group, and the difference was statistically significant ( P<0.001). The IL-6 concentration of the AICAR group was higher than that of the spore control group, but the difference was not statistically significant ( P=0.260). The IL-6 concentration of the Compound C group was lower than that of the spore control group, and the difference was statistically significant ( P=0.010). Conclusions:In corneal epithelial cells, AMPK phosphorylation is found, which is enhanced after fungal spores stimulation, and the secretion of IL-6 increases.

Purpose To determine whether Activated 5'AMP-activated protein kinase (AMPK) activation enhances Fusarium solani (F. solani) fungicidal capacity of neutrophils.Methods The expression of AMPK and phosphorylated-AMPK (p-AMPK) proteins was tested using Western Blot. Plate counting studied the effects of the fungicidal capacity of neutrophils enhanced by AMPK activation. Phagocytized spores by neutrophils were assessed by immunostaining and inhibited hyphal growth images were captured by JULI Stage real-time cell history recorder. Flow cytometry assay tested Reactive Oxygen Species (ROS) production and the percentage of apoptosis neutrophils. ROS Assay Kit also tested ROS production at different time points. The F. solani keratitis murine model was established, and slit-lamp microscopy captured corneal photographs.Results Our experiments were divided into the following groups. Neutrophils (N), neutrophils + spores (N + S), neutrophils + spores+ 5-aminoimidazole-4-carboxamide ribonucleotide (AICAR) (N + S + A), neutrophils + spores + Compound C (N + S + C). AMPK activator AICAR significantly increased the expression of p-AMPK in neutrophils. The plate counting experiment showed that the number of colonies in the N + S + A was significantly less than in the N + S group. Immunostaining results showed phagocytized spores were significantly increased in the N + S + A group compared with the N + S group. Captured photographs by a real-time cell history recorder camera showed F. solani hyphal growth in the N + S + A group was significantly inhibited than in the N + S group. ROS release in the N + S + A group was significantly higher in the N + S + A group than in other groups. The percentage of apoptosis neutrophils in the N + S + A group was decreased than in the N + S group. Captured photographs by slit-lamp showed that AICAR eye drop treatment alleviated the severity and decreased clinical score at 12 and 24 hours post-infection (h.p.i)Conclusion AMPK activation enhances the efficacy of neutrophils in killing F. solani in vitro and in vivo.

Objectie To investigate the susceptibility of drug-resistant staphylococci isolated from different parts of the anterior segment to levofloxacin, tobramycin, cefazolin sodium, fusidic acid and clindamycin. Methods: Experimental Study. A total of 67 patients with anterior segment infection (33 cases of conjunctivitis, 6 cases of bacterial keratitis, 7 cases of blepharitis, 9 cases of neonatal dacryocystitis, 9 cases of neonatal dacryocystitis, 1 case of adult dacryocystitis and 11 cases of other infectious eye diseases) were collected from the conjunctival sac, cornea, eyelid margin and lacrimal sac. Minimum inhibitory concentration (MIC) determination of methicillin-resistant Staphylococcus (MRS) strains and β-lactamase-producing (β-Lac) strains by a micro-liquid-based method, according to the M100 standard of the American Institute for Clinical and Laboratory Standardization Susceptibility and resistance determinations were made. Data were statistically analyzed using Chi-square or Fisher's exact test. Results: Thirty-five MRS, 30 β-Lac and 2 β-Lac MRS isolates were identified from 67 multidrug-resistant Staphylococcus . There were 3, 9, 4, and 19 MRS isolates isolated from the lacrimal sac, cornea, eyelid margin and conjunctival sac, accounting for 3/4, 9/12, 4/8, 19/43 (44.2%) of the isolated sites respectively. There were 1, 3, 3, and 23 β-Lac isolates, accounting for 1/4, 3/12, 3/8 and 23/43 (53.5%) of the isolated sites, respectively. The highest proportion of β-Lac isolates isolated from patients with a diagnosis of conjunctivitis was 17 (25.3%) from the conjunctival sac. Among the MRS strains isolated from the cornea and lacrimal sac, 5 (7.5%) and 3 (4.5%) were from patients diagnosed with bacterial keratitis and neonatal tear, respectively. The number of MRS strains and β-Lac isolates isolated from patients with a diagnosis of blepharitis were both 3 (4.5%) from the lid margin.Among the strains isolated from the eyelid margin and the conjunctival sac, drug-resistant Staphylococcus epidermidis was the main strain, the drug-resistant Staphylococcus aureus was the major isolates in lacrimal sac and cornea. Among the 35 MRS isoaltes, 25, 24, 12, 12, and 11 were sensitive to cefazolin sodium, fusidic acid, levofloxacin, clindamycin and tobramycin, and the sensitivity rates were 71.4%, 68.6%, 34.3%, 34.3% and 31.4%, the difference was statistically significant (χ2=22.756, P<0.001), The sensitivity rates of levofloxacin, tobramycin, cefazolin sodium, fusidic acid and clindamycin against MRS isolates from the anterior segment were both statistically significant differences (χ2=18.493, 11.594, 8.906, 9.841, 16.059; all P<0.05). The susceptibility rates of MRS isolates against five antibiotics was statistically significant differences (χ2=33.080, P<0.001). Among the 30 β-Lac isolates, 27, 22, 19, 16, and 8 were sensitive to cefazolin sodium, fusidic acid, levofloxacin, tobramycin and clindamycin, and the sensitivity rates were 90.0 % , 73.3%, 63.3%, 53.3% and 26.7%, the difference was statistically significant (χ2=28.280, P<0.001). The sensitivity rates of five antibiotics against β-Lac isolates from the anterior segment were both statistically significant differences (χ2=50.971, 24.543, 48.147, 44.899, 18.676; all P<0.001). The susceptibility rates of β-Lac isolates against five antibiotics was statistically significant differences (χ2=23.383, P<0.001). The sensitivity of cefazolin sodium and fusidic acid against β-Lac isolates were higher than MRS isolates. Conclusions: Cefazolin sodium and fusidic acid may be the best choice for the treatment of drug-resistant Staphylococcus isolated from anterior conjunctival sac, cornea, eyelid margin and lacrimal sac, especially for β-Lac-producing drug-resistant Staphylococcus infection.

目的:研究血小板对中性粒细胞体外抗真菌作用的影响.方法:实验真菌为腐皮镰胞菌标准株.分别分离健康人外周血中的中性粒细胞(N)和血小板(PL).预先分别用钙荧光白、抗人CD15-FITC和抗人CD41a-PE标记真菌孢子(S)、中性粒细胞和血小板.将中性粒细胞5×105个/孔均匀接种于培养板.①设定S+N组、S+N+PL组,按照S:N:PL个数1:1:20接种,分别于培养1、2、4 h后测定中性粒细胞吞噬率.②设置S组、S+N组、S+N+PL组,按照S:N:PL个数比例为1:5:100接种;分别于培养1、2、4 h后测定孢子出芽率;分别于培养4、6、8 h后测量菌丝伸长度.③设置N组、S+N组、S+N+PL组,按照S:N:PL个数为1:5:100接种,分别于培养6、10、14、18 h后,Annexin V/PI双染法检测中性粒细胞凋亡率.结果:S+N+PL组中性粒细胞吞噬率大于S+N组(P<0.05).S+N+PL组孢子出芽率、菌丝伸长度小于S+N组,S+N组小于S组(P<0.05).S+N+P组中性粒细胞凋亡率小于N组和S+N组(P<0.05).结论:血小板可体外增强中性粒细胞的抗真菌作用.

一例误诊的角膜异物病例,初次共聚焦显微镜检查未能提供诊断依据,通过联用几种裂隙灯显微镜检查方法,仔细反复检查发现了异物影,再到角膜刮片镜检确诊了角膜异物.回顾病史并再次察看既往拍摄的共聚焦显微镜图片,发现已拍摄到角膜异物影,因操作者经验不足而见而不识.强调需结合疾病特点和患者情况,联合使用相关检查.

Objective:To study the antimicrobial activity of diacerein on common pathogens of the ocular surface in vitro.Methods:Pathogens were collected from patients with ocular surface infections in Henan Eye Hospital, including Gram-positive cocci and bacilli, Gram-negative bacilli, filamentous fungi, and Candida.The antimicrobial activity of diacerein was determined by the K-B agar diffusion method, and its minimum inhibitory concentration (MIC) was determined by the micro-liquid method.Levofloxacin and voriconazole were used as the control of antibacterial and antifungal drug, respectirely.Results:Diacerein showed antibacterial activity against 42 strains of Gram-positive cocci and 10 strains of Gram-positive bacilli, its inhibition zone diameters for Staphylococcus epidermidis, S.aureus, S.intermedius and Gram-positive Corynebacterium were not significantly different from those of levofloxacin (all at P>0.05). Its MIC range of diacetate against Staphylococcus epidermidis, S. aureus, S. intermedius and other Staphylococci was 1-32 μg/ml, and its respective MIC 90 was 16, 8, 16, and 32 μg/ml.Diacerein had no bacteriostatic effect on 23 strains of Gram-negative bacilli, 10 strains of filamentous fungi and 3 strains of candida. Conclusions:Diacerein has antibacterial effects against Gram-positive Staphylococcus and Corynebacterium isolated from the ocular surface, but shows no antimicrobial activity against Gram-negative bacilli and fungi.Diacerein offers a new drug option and method for the treatment of bacterial keratitis.

AMP依赖的蛋白激酶(AMPK)是调节能量变化的关键因子,当体内的AMP/ATP比例升高时AMPK激活,使细胞由合成代谢转向分解代谢,能量状态恢复平衡.近年来研究发现,AMPK亦参与调控感染性疾病.本文主要阐述AMPK在感染性疾病(细菌、病毒、寄生虫、真菌)中的防治作用.AMPK通过抑制病原体的胞内生长,增强宿主对病原体的杀灭能力,介导宿主抗炎,促进宿主细胞存活而发挥作用,为感染性疾病的防治、新型抗感染药物研发提供新的思路.

Objective: To investigate the in vitro antibacterial sensitivity of levofloxacin, tobramycin, cefazolin sodium, clindamycin and fusidic acid to 67 strains of Staphylococci in ocular surface infection. The purpose of this study is to provide reference for clinical selection of drugs. Methods: Experimental study. Sixth-seven strains of drug-resistant Staphylococci isolated from the Department of Microbiology, Henan Provincial Ophthalmic Hospital during January 2018 and May 2019 were collected. There were 67 strains of Staphylococci including 28 strains of drug-resistant Staphylococcus epidermidi, 17 strains of drug-resistant Staphylococcus aureus, 15 strains of drug-resistant Staphylococcus intermedius and a few other kinds of drug-resistant Staphylococci. The minimum inhibitory concentrations (minimum inhibitory concentration, MIC) of levofloxacin, tobramycin, cefazolin sodium, clindamycin and fusidic acid in 67 strains of drug-resistant Staphylococci were determined by microliquid-based method. The sensitivity was determined according to the American CLSI-M100 standard. The statistical analysis of the data was carried out by using two-dimensional test and Fisher accurate test. Results: Fourteen strains of fusidic acid were sensitive to 17 strains of MRS-Meca-resistant Staphylococcus epidermidis, the difference between fusidic and levofloxacin is statistically significant; 14 strains of cefazolin sodium and 11 strains of fusidic acid were sensitive to 14 strains of β-Lac enzyme-producing Staphylococcus aureus, and there were significant differences between the two drugs and levofloxacin; 6 strains of cefazolin sodium and 5 strains of fusidic acid were sensitive to 10 strains of MRS-Meca-resistant Staphylococcus intermedius, as compared to levofloxacin, there were significant differences between cefazolin sodium and levofloxacin (P=0.011,0.033). Cefazolin sodium was sensitive to 5 strains of MRS-Meca-positive other drug-resistant Staphylococci, which was significantly different from levofloxacin (P=0.048); 54 and 48 strains of cefazolin sodium and fusidic acid were sensitive to 67 strains of drug-resistant Staphylococci, and the sensitive rates were 80.1% and 71.6%, respectively, which were significantly higher than those of levofloxacin, tobramycin and clindamycin. There were significant statistical differences between drug sensitivity (χ²=18.377,9.940；P=0.000,0.003). Conclusions: The sensitivity of cefazolin sodium and fusidic acid to 67 strains of drug-resistant Staphylococci is better than that of levofloxacin, tobramycin and clindamycin, these findings may provide guidance for the clinical treatment of drug-resistant Staphylococci in ocular infection. (Chin J Ophthalmol, 2020, 56: 621-625).

Objective To study the effects of platelets on macrophages phagocytosis and inhibition of fungi. Methods Macrophages were cultured, Fusarium Pythium spores were extracted and platelets were isolated from blood of mouse.Simple spore group, spore+ macrophage group and mixed platelet group were set, and were inoculated with fungal spore, equal proportion spore+ macrophage and platelet+ spore+ macrophage, respectively.The prepared plate was placed on a spinning disk laser scanning confocal microscope at 1 hour, 2, 3 and 4 hours after culture, five visual fields were randomly selected at the corresponding time points for photography.The phagocytic rate, phagocytic index and fungal spore germination rate were calculated.Fungal hyphae length of each group at 4, 6 and 8 hours after culture were calculated.The single macrophage group, spore+ macrophage group and mixed platelet group were set and the cytotoxicity was measured by real-time cell analyzer.The breeding and use of mice was in according with the ARVO statement.This study was approved by the Experimental Animal Ethics Committee of Henan Eye Institute (HNEECA-2017-04). Results The phagocytic rate and phagocytic index of macrophages in mixed platelet group at 1 hour, 2, 3 and 4 hours after culture were significantly higher than those in spore+ macrophage group at corresponding time point (all at P 0.05). There were significant differences in cell index among different groups at 30, 36, 42 and 48 hours after culture (F=10.81, 8.08, 5.61, 5.72; all at P<0.05). The cell indexes in spore+ macrophage group at different time points were significantly lower than those in simple macrophage group (all at P<0.01). Conclusions Platelets can promote the phagocytosis and inhibition of macrophages on fungi, and platelets may have antagonistic effect on fungal cytotoxicity. Key words: Keratitis/pathology; Macrophage; Fungi; Platelets; Phagocytosis; Germination rate

Objective To study the response of neutrophils of fungal keratitis in mice under different light conditions.Methods 93 male SPF C57BL/6 mice aged 6-8w were selected and bred under the following light conditions,LD group mice received light from 7:00 to 19:00 (light intensity 110-210IW/cm2),LL group received 24h light (light intensity 110 -210IW/cm2),DD group received 24h dark, all mice were bred under LD environment for 7 days,the mice were divided into three groups, LD group, LL group and DD group, the mice were fed for 12-15 days under the corresponding light conditions. To make fungal keratitis model, among which 12 mice were only used for clinical observation, respectively 0h before modeling, 12h,18h,24h,36h,48h,72h,120h,168h after modeling, and slit lamp microscope was used for anterior segment photography and clinical scoring.The rest of the 81 mice was performed after the above time point executed based respectively,immunofluorescence staining was performed on the whole cornea,using a single photon laser confocal microscope,the whole corneal film was scanned under 40x oil lens,the infiltration of neutrophils in cornea of mice treated at different time points and under different experimental conditions was calculated by using image processing software. Results The total keratitis score of LD group was higher than that of DD group,at 24 h after modeling, the clinical score of corneal inflammation of LD group was significantly higher than that of DD group (P=0.008).The total keratitis score of LL group was higher than that of DD group,at 18h, 24h, 36h and 168h after modeling ,the clinical scores of corneal inflammation of LL group were significantly higher than those of DD group (P=0.013, 0.004, 0.008, 0.022 < 0.05),at 12 h and 48 h after modeling, the infiltration of neutrophils in the cornea of LD group was significantly higher than that of DD group (P=0.033, 0.009 < 0.05).The infiltration of neutrophils in the cornea of LL group was higher than that of DD group,at 12h, 24h and 48h after modeling, the infiltration of neutrophils in the cornea of LL group was significantly higher than that of DD group (P=0.002, 0.022, 0.001 < 0.05).The infiltration of neutrophils in the three groups was positively correlated with clinical scores (r = 0.192, 0.454, P = 0.019, 0.000).Conclusion In the stage of fungal infection in mice, light helps to increase the recruitment of neutrophils ,more neutrophils infiltrated, leading to more severe inflammation of fungal keratitis in mice.

Bacterial keratitis is an aggressive infectious corneal disease. With the continuing rise in antibiotic resistance and a decline in the discovery of new antibiotics, new antimicrobial drugs are now required. In the present study, we determined the antibacterial activity of diacerein, an anti-inflammatory drug, against 76 Gram-positive cocci isolated from bacterial keratitis patients in vitro and anti-Staphylococcus aureus activity in a mouse bacterial keratitis model in vivo The MICs of diacerein were tested using the broth microdilution method in vitro A BALB/c Staphylococcus aureus keratitis animal model was selected and the corneal clinical observation, viable bacteria, and hematoxylin-eosin and Gram staining of infected corneas were measured to evaluate the antibacterial efficacy of diacerein eye drops in vivo An in vivo eye irritation study was carried out by a modified Draize test in rabbits. Our in vitro results showed that diacerein possesses satisfactory antibacterial activity against the majority of Gram-positive cocci (60/76), including all 57 tested Staphylococcus spp. and 3 Enterococcus spp. The in vivo experiment showed that diacerein eye drops reduced bacterial load and improved ocular clinical scores after topical administration of diacerein drops on infected corneas. The ocular irritation test revealed that diacerein eye drop had excellent ocular tolerance. These results indicated that diacerein possesses in vivo anti-Staphylococcus aureus activity. We suggest that diacerein is a possible topically administered drug for Staphylococcus aureus-infected patients, especially those with ocular surface inflammatory disorders.