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Comparison of four supports for adsorption of reactive dyes by immobilized aspergillus fumigatus beads

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Journal of Environmental Sciences 2007年第4期19卷 451-457页

作者： WANG Bao-e HU Yong-you School of Environmental Science and Engineering South China University of Technology Guangzhou 510640 China.

Four materials, sodium carboxymethylcellulose （Na-CMC）, sodium alginate （SA）, polyvinyl alcohol （PVA）, and chitosan （CTS）, were prepared as supports for entrapping fungus aspergillusfumigatus. The adsorption of synthetic dyes, Reactive Brilliant Blue KN- R, and Reactive Brilliant Red K-2BP, by these immobilized gel beads and plain gel beads was evaluated. The adsorption efficiencies of Reactive Brilliant Red K-2BP and Reactive Brilliant Blue KN-R by CTS immobilized beads were 89.1% and 93.5% in 12 h, respectively. The adsorption efficiency by Na-CMC immobilized beads was slightly lower than that of mycelial pellets. But the dye culture mediums were almost completely decolorized in 48 h using the above-mentioned two immobilized beads （exceeding 95%）. The adsorption efficiency by SA immobilized beads exceeded 92% in 48 h. PVA-SA immobilized beads showed the lowest adsorption efficiency, which was 79.8% for Reactive Brilliant Red K-2BP and 92.5% for Reactive Brilliant Blue KN-R in 48 h. Comparing the adsorption efficiency by plain gel beads, Na-CMC plain gel beads ranked next to CTS ones. SA and PVA-SA plain gel beads hardly had the ability of adsorbing dyes. Subsequently, the growth of mycelia in Na-CMC and SA immobilized beads were evaluated. The biomass increased continuously in 72 h. The adsorption capacity of Reactive Brilliant Red K-2BP and Reactive Brilliant Blue KN-R by Na-CMC immobilized beads was 78.0 and 86.7 mg/g, respectively. The SEM micrographs show that the surface structure of Na-CMC immobilized bead is loose and finely porous, which facilitates diffusion of the dyes.

关键词： adsorption aspergillus fumigatus immobilization reactive dyes

TREM-1 expression in rat corneal epithelium with aspergillus fumigatus infection

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International Journal of Ophthalmology(English edition) 2015年第2期8卷 222-227页

作者： Li-Ting Hu Zhao-Dong Du Gui-Qiu Zhao Sheng Qiu Nan Jiang Jing Lin Qian Wang Qiang Xu Department of Ophthalmology Affiliated Hospital of Qingdao University

AIM: To investigate the expression of triggering receptor expressed on myeloid cells-1(TREM-1) in the aberrant inflammation within the corneal epithelium at early period of fungal ***: A total of 65 Wistar rats were randomly divided into control group, sham group and fungal keratitis(FK) group, in which the cornea was infected by aspergillus fumigatus(A. fumigatus). After executed randomly at 8, 16, 24, 48 and 72 h after experimental model being established, the severity of keratomycosis in rats was scored visually with the aid of a dissecting microscope and slit lamp. Then corneas in three groups were collected to assess the expression of TREM-1through quantitative reverse transcription-polymerase chain reaction(RT-PCR), immunofluorescence technique and Western blot analysis. The correlation between FK inflammation and expression of TREM-1 was also ***: Corneal inflammation scores increased with time after fungal infection(F =49.74, P =0.000). The inflammation scores in FK group were obviously higher than those in sham group on the whole(F =137.78, P =0.000). Levels of TREM-1 in the infected rat corneal epithelium had elevated at 8h and peaked at 48h(P fumigatus and play critical roles in the battle against A. fumigatus in the innate immune responses.

关键词： fungal keratitis triggering receptor expressed on myeloid cells-1 aspergillus fumigatus corneal epithelium

Design,synthesis of novel antifungal triazole derivatives with high activities against aspergillus fumigatus

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Chinese Chemical Letters 2007年第4期18卷 421-423页

作者： Qiu Qin He Chao Mei Liu Ke Li Yong Bing Cao School of Pharmacy Second Military Medical University Shanghai 200433 China

Based on the active site of aspergillus fumigatus lanosterol 14α-demethylase （AF-CYP51）, novel triazole compounds were designed. Their chemical synthesis and the antifungal activities were reported. The results showed that all the target compounds exhibited excellent activities with broad spectrum; in which compounds 4, 12 and 15 showed comparable activities against A. fumigatus to the control drug itraconazole.

关键词： CYP51 Antifungal Triazole Synthesis aspergillus fumigatus

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

Thymol Protects against aspergillus fumigatus Keratitis by Inhibiting the LOX-1/IL-1βSignaling Pathway

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Current Medical Science 2022年第3期42卷 620-628页

作者： Li-mei WANG Hua YANG Hai-jing YAN Rui-feng GE Yun-xiao WANG Sha-sha XUE Lin LI Le-yu LYU Cheng-ye CHE Department of Ophthalmology the Affiliated Hospital of Qingdao UniversityQingdao 266003China Department of Otorhinolaryngology-Head and Neck Surgery the Affiliated Hospital of Qingdao UniversityQingdao 266003China

Objective To explore the anti-inflammatory effects and mechanisms of action of thymol in aspergillus fumigatus(***)*** The minimum inhibitory concentration of thymol against *** was *** characterize the anti-inflammatory effects of thymol,mouse corneas and human corneal epithelial cells were pretreated with thymol or dimethyl sulfoxide(DMSO)before infection with *** ***-lamp microscopy,immunohistochemistry,myeloperoxidase detection,quantitative real-time polymerase chain reaction,and Western blotting were used to assess *** and macrophage recruitment,in addition to the secretion of LOX-1 and IL-1β,were quantified to evaluate the relative contribution of thymol to the inflammatory *** We confirmed that the growth of *** was directly inhibited by *** contrast with the DMSO group,there was a lower degree of inflammation in the mouse corneas of the thymol-pretreated *** was characterized by significantly lower clinical scores,less inflammatory cell infiltration,and lower expression of LOX-1 and IL-1β.Similarly,in vitro experiments indicated that the production of LOX-1 and IL-1βwas significantly inhibited after thymol treatment,in contrast with the DMSO-pretreated *** Our findings demonstrate that thymol exerted a direct fungistatic activity on ***,thymol played a protective role in fungal keratitis by inhibiting LOX-1/IL-1βsignaling pathway and reducing the recruitment of neutrophils and macrophages.

关键词： keratitis aspergillus fumigatus thymol LOX-1 IL-1β

Dectin-1 agonist curdlan modulates innate immunity to aspergillus fumigatus in human corneal epithelial cells

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International Journal of Ophthalmology(English edition) 2015年第4期8卷 690-696页

作者： Cheng-Cheng Zhu Gui-Qiu Zhao Jing Lin Li-Ting Hu Qiang Xu Xu-Dong Peng Xue Wang Sheng Qiu Department of Ophthalmology the Affiliated Hospital of Qingdao University

· AIM: To explore the immunomodulatory effects of curdlan on innate immune responses against aspergillus fumigatus(A. fumigatus) in cultured human corneal epithelial cells(HCECs), and whether C-type lectin receptor Dectin-1 mediates the immunomodulatory effects of curdlan.·METHODS: The HCECs were stimulated by curdlan in different concentrations(50, 100, 200, 400 μg/m L) for various time. Then HCECs pretreated with or without laminarin(Dectin-1 blocker, 0.3 mg/m L) and curdlan were stimulated by A. fumigatus hyphae. The m RNA and protein production of tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6) were determined by real-time quantitative polymerase chain reaction and enzyme-linked immunosorbent assay, respectively. The protein level of Dectin-1 was measured by Western blot.· RESULTS: Curdlan stimulated m RNA expression of TNF-α and IL-6 in a dose and time dependent manner in HCECs. Curdlan pretreatment before A. fumigatus hyphae stimulation significantly enhanced the expression of TNF-α and IL-6 at m RNA and protein levels compared with A. fumigatus hyphae stimulation group(P fumigatus hyphae up-regulated Dectin-1 protein expression in HCECs, and Dectin-1expression was elevated to 1.5- to 2-fold by curdlan pretreatment followed hyphae stimulation. The Dectin-1blocker laminarin suppressed the m RNA expression and protein production of TNF-α and IL-6 induced by curdlan and hyphae(P fumigatus hyphae in ***-1 is essential for the immunomodulatory effectsof curdlan. Curdlan may have high clinical application values in fungal keratitis treatment.

关键词： curdlan Dectin-1 innate immunity aspergillus fumigatus corneal epithelium

CCPG1 involved in corneal aspergillus fumigatus infection

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International Journal of Ophthalmology(English edition) 2022年第4期15卷 541-546页

作者： Li-Mei Wang Xiao-Meng Chen Hai-Jing Yan Shu Yan Xiao-Yan Sun Da-Wei Zhang Hua Yang Dan-Li Lu Cheng-Ye Che Department of Ophthalmology the Affiliated Hospital of Qingdao UniversityQingdao 266000Shandong ProvinceChina Department of Otorhinolaryngology-Head and Neck Surgery the Affiliated Hospital of Qingdao UniversityQingdao 266000Shandong ProvinceChina Department of Emergency the Affiliated Hospital of Qingdao UniversityQingdao 266000Shandong ProvinceChina

AIM: To investigate whether non-canonical autophagy transport receptor cell cycle progression 1(CCPG1) is involved in the corneal antifungal immune ***: Human corneal epithelial cells(HCECs) and human myeloid leukemia mononuclear cells(THP-1) macrophages stimulated by Aspergil us fumigatus(A. fumigatus) were used as cell models. The expression of CCPG1 m RNA was detected by q RT-PCR. Western blot was used to determine the protein expression of CCPG1 and interleukin-1β(IL-1β). The dectin-1 neutralizing antibody was used to detect the association between dectin-1 and CCPG1. Immunofluorescence was used to observe the colocalization of CCPG1 and C-type lectin-like receptor-1(CLEC-1) in THP-1 ***: The expression of CCPG1 started to increase at 4 h after infection and increased in a time-dependent manner in HCECs and THP-1 macrophages. With dectin-1 neutralizing antibody pretreatment, the expression of IL-1β was down-regulated. CCPG1 up-regulation in response to A. fumigatus infection was independent of dectin-1. Immunofluorescence showed the colocalization of CCPG1 and CLEC-1 in THP-1 ***: As a specific autophagy protein of noncanonical autophagy pathway, CCPG1 is involved in corneal infection with A. fumigatus.

关键词： cell cycle progression 1 fungal keratitis aspergillus fumigatus C-type lectin-like receptor-1

Interactions of thymic stromal lymphopoietin with interleukin-4 in adaptive immunity during aspergillus fumigatus keratitis

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International Journal of Ophthalmology(English edition) 2021年第10期14卷 1473-1483页

作者： Chen Chen Chen-Yang Dai Fang Han Jia-Yin Wu Lin Sun Xin-Yi Wu Department of Ophthalmology Clinical Medical College of Shandong UniversityJinan 250012Shandong ProvinceChina Department of Ophthalmology Qilu Hospital of Shandong UniversityJinan 250012Shandong ProvinceChina Key Laboratory of Cardiovascular Remodeling and Function Research Shandong UniversityJinan 250012Shandong ProvinceChina

AIM:To investigate the potential interactions of thymic stromal lymphopoietin(TSLP)with interleukin-4(IL-4)in adaptive immunity during fungal keratitis(FK).METHODS:An FK mouse model was induced with aspergillus fumigatus(AF)hyphal *** were divided into several groups:untreated,phosphate buffer saline(PBS),infected with AF,and pretreated with a scrambled siRNA,a TSLP-specific siRNA(TSLP siRNA),murine recombinant TSLP(rTSLP),immunoglobulin G(IgG),murine recombinant IFN(rIFN-γ),murine recombinant IL-4(rI L-4),rIL-13,murine recombinant IL-17A(rIL-17A),and murine recombinant IL-17F(rIL-17F)*** realtime reverse transcription-polymerase chain reaction(qRTPCR)and enzyme-linked immunosorbent assay(ELISA)or Western blot were performed to determine mRNA and protein levels in the inflamed *** locations were observed by immunofluoresence staining after AF hyphal ***:Compared to those in the untreated group,TSLP and T helper type 1(Th1)cytokine levels in the AF group were upregulated at 24 h post infection(hpi),and those of T helper type 2(Th2)and T helper type 17(Th17)cytokines were increased at 5 d post infection(dpi).Th2 cytokine levels were decreased in the TSLP siRNA-pretreated group and increased in the rTSLP-pretreated group compared with the AF *** TSLP level was increased in the rIL-4-pretreated group,but there were no significant changes among the other *** staining showed cytokine locations after AF hyphal ***:TSLP induces a Th2 immune response and promots Th2 T cell differentiation in ***-4 promotes TSLP ***,TSLP with IL-4 regulates adaptive immunity in FK.

关键词： aspergillus fumigatus keratitis thymic stromal lymphopoietin Th2 immune response interleukin-4

The role of Dectin-1/Raf-1 signal cascade in innate immune of human corneal epithelial cells against aspergillus fumigatus infection

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International Journal of Ophthalmology(English edition) 2016年第10期9卷 1371-1375页

作者： Gui-Qiu Zhao Jing Lin Li-Ting Hu Xiao-Ni Yin Qian Wang Qiang Xu Hui Li Department of Ophthalmology the Affiliated Hospital of Qingdao University

AIM： To investigate the expression of the v-raf-1murine leukemia viral oncogene homolog 1（Raf-1） and its role in the innate immune response of human corneal epithelial cells（HCECs） infected by aspergillus ***： HCECs were cultured in *** were randomly divided into 4 groups,including control group,aspergillus fumigatus group,GW5074（an inhibitor of Raf-1） group and Laminarin [an inhibitor of Dendriti-cell-associated C-type lectin 1（Dectin-1）] *** protein expression level of total Raf-1 and p-Raf-1 was measured by Western *** expression of IL-6 and IL-8 m RNA in each group was detected by real-time polymerase chain ***： In aspergillus fumigatus group,total Raf-1 protein levels in HCECs remained unchanged at 5,15,30 and 45min after infection,while p-Raf-1 expression was significantly enhanced at 30 min after infection compared with control ***,the expression of p-Raf-1 was apparently declined after treated with GW5074 or Laminarin compared with aspergillus fumigatus *** expression levels of IL-6,IL-8 m RNA were significantly increased after stimulation with fumigatus compared with control ***-treated with GW5074 significantly inhibited aspergillus fumigatus-induced upregulation of IL-8 and ***： aspergillus fumigatus stimulation can elevate the expression of p-Raf-1 in HCECs in ***-1/Raf-1 signal pathway may play a role on regulating the expression of inflammatory cytokines,including IL-6 and IL-8.

关键词： Dectin-1/Raf-1 signal pathway aspergillus fumigatus innate immune human corneal epithelial cells