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Comparison of cytotoxicity and wound healing effect of carboxymethylcellulose and hyaluronic acid on human corneal epithelial cells

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International Journal of Ophthalmology(English edition) 2015年第2期8卷 215-221页

作者： Jong Soo Lee Seung Uk Lee cheng-ye che Ji-Eun Lee Department of Ophthalmology School of MedicinePusan National University Biomedical Research Institute Pusan National University Hospital Department of Ophthalmology School of Medicine Kosin University Department of Ophthalmology the Affiliated Hospital of Medical College Qingdao University Research Institute for Convergence of Biomedical Science and Technology Pusan National University Yangsan Hospital

AIM: To investigate the cytotoxic effect on human corneal epithelial cells(HCECs) and the ability to faciliate corneal epithelial wound healing of carboxymethylcellulose(CMC) and hyaluronic acid(HA).METHODS: HCECs were exposed to 0.5% CMC(Refresh plus, Allergan, Irvine, California, USA) and 0.1% and 0.3%HA(Kynex , Alcon, Seoul, Korea, and Hyalein mini,Santen, Osaka, Japan) for the period of 30 min, and 4, 12,and 24 h. Methyl thiazolyl tetrazoiun(MTT)-based calorimetric assay was performed to assess the metabolic activity of cellular proliferation and lactate dehydrogenase(LDH) leakage assay to assess the cytotoxicity. apoptotic response was evaluated with flow cytometric analysis and fluorescence staining with Annexin V and propiodium iodide. Cellular morphology was evaluated by inverted phase-contrast light microscopy and electron microscopy. The wound widths were measured 24 h after confluent HCECs were scratch ***: The inhibitory effect of human corneal epithelial proliferation and cytotoxicity showed the time-dependent response but no significant effect. Apoptosis developed in flow cytometry and apoptotic cells weredemonstrated in fluorescent micrograph. The damaged HCECs were detached from the bottom of the dish and showed the well-developed vacuole formations. Both CMC and HA stimulated reepithehlialization of HCECs scratched, which were more observed in ***: CMC and HA, used in artificial tear formulation, could be utilized without any significant toxic effect on HCECs. Both significantly stimulated HCEC reepithelialization of corneal wounds.

关键词： carboxymethylcellulose corneal wound healing human corneal epithelial cells hyaluronic acid cytotoxicity

Early expression of PTX3 in Aspergillus fumigatus infected rat cornea

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International Journal of Ophthalmology(English edition) 2018年第7期11卷 1084-1089页

作者： Jie Zhang Gui-Qiu Zhao Jing Qu Jing Lin cheng-ye che Xue-Jiao Yang Department of Ophthalmology the Affiliated Hospital of Qingdao University Department of Administrative Office the Affiliated Hospital of Qingdao University

AIM： To investigate the expression of pentraxin 3（PTX3） in rat corneal epithelium at the early stage of Aspergillus fumigatus（A. fumigatus） infection. METHODS： A total of 50 Wistar rats were randomly divided into control group, Sham group and experimental group（fungal keratitis group, FK group）. The right eye was chosen as the experiment one and infected by A. fumigatus. Rats were executed at 8, 16 and 24 h after the experimental models being established. Corneal epithelia were collected to assess the expression of PTX3 by quantitative reverse transcription polymerase chain reaction（q RT-PCR） and Western blot analysis. RESULTS： Corneal inflammation scores increased as infection prolonged（P〈0.05, P〈0.001）. PTX3 m RNA expression was low in normal and Sham group rats＇ corneas. Level of PTX3 m RNA in infected rat cornea was elevated at 8 h and peaked at 16 h. The difference was significant compared with control group（P〈0.001）. Western blot analysis also showed a significant increase of PTX3 protein in experimental group at 8 h and peaked at 16 h（P〈0.001）. The synchronous expression of control group and experimental group were also in significant difference（P〈0.001）. CONCLUSION： PTX3 exists in cornea epithelium and is significantly increased after A. fumigatus infection. PTX3 plays an important role in the early stage of cornea innate immunity against A. fumigatus.

关键词： fungal keratitis AspergiIlus fumigates pentraxin 3 innate immunity

Production of interleukin-1β related to mammalian target of rapamycin/Toll-like receptor 4 signaling pathway during Aspergillus fumigatus infection of the mouse cornea

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International Journal of Ophthalmology(English edition) 2018年第5期11卷 712-718页

作者： Rui Xu Jing Lin Gui-Qiu Zhao Cui Li cheng-ye che Qiang Xu Min Liu Department of Ophthalmology the Affiliated Hospital of Qingdao University

AIM：To elucidate the effect of rapamycin on regulating the production of interleukin（IL）-1β in Aspergillus fumigatus（***）-induced keratitis and to verify whether the expression of IL-1β in *** keratitis is associated with the mammalian target of rapamycin（mT OR）/Toll-like receptor 4（TLR4） signaling ***：Fungal keratitis mouse models of susceptible C57 BL/6 mice were established using *** mice were subsequently treated with *** protein levels of p-mT OR,TLR4,and IL-1β in normal and infected corneal tissue were measured by Western *** TLR4 and IL-1β m RNA levels were determined by real-time polymerase chain reaction（PCR）.RESULTS：In C57 BL/6 mice,rapamycin treatment decreased the clinical scores and production of the pro-inflammatory cytokine,IL-1β.The expression of TLR4,stimulated by ***,was reduced as well when the mT OR signaling pathway was suppressed by ***：Rapamycin is beneficial for the outcome of fungal keratitis and has an inhibitory effect expression of the inflammatory cytokine IL-1β.The inhibitory effect on IL-1β expression can be associated with the mT OR/TLR4 signaling pathway in *** infection in mice.

关键词： keratitis interleukin-1β mammalian target of rapamycin Toll-like receptor 4 mice

Effects of COX-2 inhibitor NS-398 on IL-10 expression in rat fungal keratitis

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International Journal of Ophthalmology(English edition) 2011年第2期4卷 165-169页

作者： Na Li cheng-ye che Li-Ting Hu Jing Lin Qing Wang and Gui-Qiu Zhao Department of Ophthalmology the Affiliated Hospital of Qingdao University Qingdao 266003 Shandong Province China

AIM: To investigate the expression of interleukin-10 (IL-10) and the effect of NS-398 (COX-2 inhibitor) on the expression of IL-10 in fungal keratitis in rats, and analyze its effects on anti-fungus immunity. METHODS: Ninety Wister rats were randomly divided into 3 groups. Group A was blank control group (10 eyes). Group B was fungal keratitis group (40 eyes). Group C was fungal keratitis group treated with NS-398 (40 eyes). PAS staining, 100g/L potassium hydroxide (KOH) smear and fungal culture confirmed the successful establishment of fungal keratitis model. After the central epithelium was scraped, Fusarium solani colonies were applied and contact lens was put on the right cornea of group B and C, and plane contact lens was put on the left cornea of control eyes. Phosphate buffered saline (PBS) eyedrops were given for group B and NS-398 eyedrops for group C. The expression of IL-10 on corneas of group B and C on the 1(st) day, 3(rd) days, 75(th) days, and 14(th) days were detected by immunohistochemistry and semi- quantitative reverse transcription- polymerase chain reaction (RT-PCR). RESULTS: Histopathologic examination showed neutrophil infiltration and severe tissue necrosis in ulcer cornea. PAS staining confirmed the existence of hyphae and spores in the superficial layer of stroma. In the blank and control groups almost no expression of IL-10 was detected at any observing points. In group B the expression of IL-10 increased at first and decreased thereafter. Its expression also showed significant difference at any observing points (P < 0.01). Compared with group B, the expression of IL-10 in group C showed no difference on the 1(st)day, decrease on the 3(rd) day, but a significant increase on the 7(th) day and 14(th) day. CONCLUSION: IL-10 takes part in the occurrence and development of fungal keratitis. NS-398 can upgrade the expression of IL-10 in fungal keratitis in the later period of the ulcer. Meanwhile, pathologic observation showed a slightly corneal

关键词： fungal keratitis immune regulation IL-10 NS-398

The expression of lacrimal androgen-binding proteins in mice Pseudomonas aeruginosa keratitis

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International Journal of Ophthalmology(English edition) 2020年第1期13卷 7-10页

作者： Le-Yu Lyu Qian Wang Qiang Xu Wen-Yi Zhao Hua Yang cheng-ye che Department of Ophthalmology the Affiliated Hospital of Qingdao University

AIM: To investigate the expression of lacrimal androgenbinding proteins(ABPs) in mice Pseudomonas aeruginosa(P. aeruginosa) ***: P. aeruginosa mice model from different gender was developed by intra-stromal injection. The expression of lacrimal ABPs in lacrimal gland specimens from P. aeruginosa keratitis mice was detected by the quantitative polymerase chain reaction(q RT-PCR). Corneal virulence was evaluated based on clinical scores. To study the mechanism of lacrimal ABPs' expression, experimental subjects were pre-treated with 4 E-BP1 inhibitor, and were used to evaluate the expression levels by q ***: Compared with control groups, the expression of ABPα, ABPη and ABPζ in lacrimal gland from P. aeruginosa keratitis mice had no meaningful changes, while ABPε and ABPδ were significantly higher at 1 d after infection. The expression of ABPδ in lacrimal gland of male mice was higher than female mice, regardless of whether or not P. aeruginosa keratitis occurred. After 4 E-BP1 inhibitor subconjunctival injection or lacrimal injection, the expression of ABPδ and ABPε has no significant change compared with the control ***: ABPδ and ABPε secreted by mice lacrimal gland may involve in the progress of alleviating the severity of corneal damage in P. aeruginosa keratitis. The expression of ABPδ and ABPε upon P. aeruginosa infection is independent of cap-dependent m RNA translation activated by 4 E-BP1.

关键词： keratitis Pseudomonas aeruginosa androgen-binding proteins lacrimal gland

Characteristics of ocular abnormalities in gout patients

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International Journal of Ophthalmology(English edition) 2013年第3期6卷 307-311页

作者： Jing Lin Gui-Qiu Zhao cheng-ye che Shan-Shan Yang Qian Wang Chang-Gui Li Department of Ophthalmology the Affiliated Hospital of Medical College of Qingdao University

·AIM: To characterize the clinical features of ocular surface in gout patients in coastal area of Shandong Province in China. ·METHODS: A total of 380 consecutive gout patients were examined from January 2011 to May 2011. According to the course of gout, patients were divided into group A (years), B (5 -10 years) and C (>10 years). Group D (control group) was consist of 50 healthy subjects. Eyelids, lateral canthus, medial canthus, palpebral conjunctiva, sclera and cornea, anterior chamber, lens, anterior vitreous were examined by slit lamp to find whether there were deposition of uric acid crystals, ocular vascular tortuosity, redness and subconjunctival hemorrhage. The ophthalmic exams of visual acuity, intraocular pressure, fundus were used to assess any gout-related eye disease. ·RESULTS: Uric acid crystals were found in 3 patients and the positions of the deposite were in corneal stroma, corneal epithelium and superficial stroma, and sclera respectively. The incidence was 0.79% . Dilatated and tortuous blood vessels in conjunctiva and sclera surface were found in 38 (23.8% ), 40 (44.0% ), 58 (45.0% ), 9 (18.0% ) patients in groups A, B, C and D, respectively. The differences between group B and D, group C and D were statistically significant( ye injury. ·

关键词： ocular abnormalities gout tophi

Nucleotide oligomerization domain 2 contributes to the innate immune response in THCE cells stimulated by Aspergillus fumigatus conidia

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International Journal of Ophthalmology(English edition) 2012年第4期5卷 409-414页

作者： Zheng-Jie Xu Gui-Qiu Zhao Qing Wang cheng-ye che Nan Jiang Li-Ting Hu and Qiang Xu Department of Ophthalmology the Affiliated Hospital of Qingdao University Medical College Qingdao 266003 Shandong Province China

AIM: To investigate the expression of nucleotide oligomerization domain 2 (NOD2) in the immortalized human corneal epithelial cell line (THCE), and its role in the innate immune response triggered by inactive Aspergillus fumigatus (Af) conidia. METHODS: The normal THCE cells were investigated as controls. After incubation with inactive Af conidia for 0.5, 2, 4, 6, and 8 hours, THCE cells were harvested, mRNA expression of NOD2 and receptor interacting protein 2 (RIP2) was detected by RT-PCR. Intracellular proteins including NOD2, NF-kappa B and proinflammatory cytokines such as TNF-alpha, IL-8, IL-6 in the cell supernatant were analyzed by ELISA. RESULTS: Our data indicate that NOD2 expressed in the normal THCE cells. After triggered by the inactive Af conidia, the expression of NOD2, RIP2 mRNA and the secretion of NOD2, NF-kappa B, TNF-alpha, IL-8, IL-6 both increased in a time-depended manner, and reached the peak point at 4, 6, 6, 4, 6, 6, 4 hours, respectively. And after pretreated with NOD2 neutralizing antibody, the expression of RIP2, NF-kappa B, TNF-alpha, IL-8 both decreased dramatically at the peak point, while the secretion of IL-6 changed little. CONCLUSION: The results of this study suggest that NOD2 exists and expresses in the THCE cells, and contributes to the innate immune responses triggered by inactive Afconidia by induction of proinflammatory cytokines such as TNF-alpha and IL-8 through the NF-kappa B pathway.

关键词： nucleotide oligomerization domain 2 corneal epithelial cell Aspergillus fumigatus innate immune response

Expression of S100B during the innate immune of corneal epithelium against fungi invasion

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International Journal of Ophthalmology(English edition) 2016年第2期9卷 191-197页

作者： Jie Zhang Gui-Qiu Zhao Jing Qu cheng-ye che Jing Lin Nan Jiang Han Zhao Xue-Jun wang Department of Ophthalmology the Affiliated Hospital of Qingdao University Qingdao 266003 Shandong ProvinceChina Department of Administrative Management the Affiliated Hospital of Qingdao University Qingdao 266003 Shandong Province China Department of Pathology the Affiliated Hospital of Qingdao University Qingdao 266003 Shandong Province China Department of Pharmacy the Second People's Hospital of Linyi Linyi 276499 Shandong Province China

AIM： To explore the expression of SIOOB in corneal epithelial cells under ,Aspergillus stimulation both in vivo and in vitro. METHODS： Immortalized human corneal epithelial cells （HCECs） were exposed to inactive #lsperg///us fumigatus （A. fumigatus） conidia at 0, 4, 8, 12, 16, and 24h respectively. The corneas of Wistar rats were exposed to active A. fumigatus at 0, 12, 24, 48h and the normal rat corneas were used for normal control. The mRNA level of S100B was evaluated by real time quantitative reverse transcription-polymerase chain reaction （qRT-PCR）. Sl00B protein expression in cornea epithelium was detected by immunohistochemical/immunocytochemical staining （IHC/ICC）. RESULTS： Histopathology revealed a significant inflammatory cell infiltration in fungal keratitis human and rat cornea. Corneal epithelial cells didn＇t express or rarely express S100B at baseline. A. fumigatus significantly induced S100B mRNA expression in cultured corneal epithelial cells in a time depended manner in vitro the mRNA began to rise significantly at 8h in vitro （P〈0.05） and continue to rise as time prolonged （P〈0.01）. in vivo S100B mRNA level was low in the normal corneas. However, it was increased in keratitis corneas from 12h after infection （P〈0.05） and reached to a peak at 24h （P〈0.001）. Immunochemistry revealed an obvious staining in fungal keratitis corneas as well as immortalized HCECs compared to the normal ones respectively, indicating an increased expression of SlOOB protein. CONCLUSION： S100B exists in corneal epithelial cells and is over-expressed under A. fumigatus stimulation. Sl00B may play an important role in the innate immune response of the corneal epithelium during A. fumigatus infection.

关键词： SIOOB cornea Aspergillus fumigates innate immunity

CD3ε of a pan T cell marker involved in mouse Aspergillus fumigatus keratitis

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International Journal of Ophthalmology(English edition) 2024年第4期17卷 616-624页

作者： Wen-Hao Shi Li-Mei Wang Hai-Jing Yan Shi-Long Liu Xian Yang Xue-JiaoYang cheng-ye che Department of Ophthalmology the Affiliated Hospital of Qingdao UniversityQingdao 266000Shandong ProvinceChina Weifang Medical University Weifang 261053Shandong ProvinceChina Eye Hospital of Wenzhou Medical University Wenzhou 325027Zhejiang ProvinceChina Department of Medical Engineering the Affiliated Hospital of Qingdao UniversityQingdao 266000Shandong ProvinceChina

AIM:To explore whether CD3ε is involved in the adaptive immunity of Aspergillus fumigatus(***)keratitis in mice and the role of innate and adaptive immunity in ***:Mice models of *** keratitis were established by intra-stromal injection and corneal epithelial *** injections of natamycin,wedelolactone,LOX-1 inhibitor(poly I)or Dectin-1 inhibitor(laminarin)were used to treat mice with *** *** were pretreated by intraperitoneal injection of anti-mouse CD3ε.We observed the corneal infection of mice under the slit lamp microscope and made a clinical *** protein expression of CD3ε and interleukin-10(IL-10)was determined by Western ***:With the disease progresses,the degree of corneal opacity and edema *** the intrastromal injection models,CD3εprotein expression began to increase significantly on the 2^(nd) ***,in the scraping epithelial method models,CD3ε only began to increase on the 3^(rd) *** natamycin treatment,the degree of corneal inflammation in mice was significantly attenuated on the 3^(rd) *** wedelolactone treatment,the severity of keratitis *** the amount of CD3ε protein was also reduced,compared with the control *** inhibiting LOX-1 and Dectin-1,there was no significant difference in CD3ε production compared with the control *** inhibiting CD3ε,corneal ulcer area and clinical score increased,and IL-10 expression was ***:As a pan T cell marker,CD3ε participate in the adaptive immunity of *** keratitis in *** our mice models,the corneas will enter the adaptive immune stage *** regulating IL-10,CD3ε exerts antiinflammatory and repairs effects in the adaptive immune stage.

关键词： CD3ε adaptive immunity Aspergillus fumigatus keratitis interleukin-10