So far, intense pulsed light (IPL) has been widely used in the treatment of meibomian gland dysfunction (MGD), but there was still a lack of research on its specific mechanism. Determining whether there was a correlation between liposome changes and remission of clinical signs in patients with MGD treated with IPL was of great significance in the clinical evaluation of efficacy in patients with MGD. Our study enrolled the 10 healthy subjects and 26 adult patients, who were diagnosed with MGD and had not received any alternative treatments for at least 3 months. Each patient received a series of three treatments at 3-week intervals. The meibum was collected before the first treatment (T0) and the third treatment (T2). The significant changes in ocular surface parameters before and after IPL treatment were analyzed. The results showed that IPL significantly improved the symptoms of MGD, including ocular surface disease index (OSDI), tear breakup time (TBUT), redness of conjunctival (CR), corneal fluorescein staining (CF), the meibomian gland expressibility (MGE), and meibum quality (all p < 0.05). Lipidomics analysis of the meibum characterized the changes in lipid profiles induced by IPL. A total of 323 lipid species compounds were identified in the spectrum. A total of 41 lipid species were significantly different in patients with MGD (T0) vs. healthy controls. Following IPL treatment (T2), 24 lipid species were significantly different compared with T0: TG (10 lipid species), LPC (6 lipid species), OAHFA (4 lipid species), Cer (2 lipid species), SM (1 lipid species), and PE (1 lipid specie). Among these lipids, 4 of the lipids was a high correlation with TBUT, 5 was TH, 6 was CR, and 11 was meibum quality. In a ward, IPL treatment can achieve the therapeutic effect by changing the alternations of tear film lipids in patients with MGD. The changes in lipid expression profiles are potential indexes to evaluate the therapeutic effectiveness of IPL treatment or other treatments on MGD.
Objective. The aims of our experiment were to compare the microorganisms in meibomian gland secretions from patients with internal hordeolum before and after treatment using hypochlorous acid eyelid wipes, to elucidate the mechanism underlying hypochlorous acid eyelid wipe treatment of internal hordeolum. Methods. This was a prospective, matched-pair study. A total of eight patients with internal hordeolum who attended the ophthalmology clinic of our hospital from April to August 2020 were included. Meibomian gland secretions were collected from subjects before treatment (Group A) and from patients cured after eyelid cleaning with hypochlorous acid eyelid wipes for 7 days (Group B). Samples were submitted to 16S rRNA high-throughput sequencing, and the resulting data were analyzed to compare the differences in the structure and composition of meibomian gland secretion microbial flora before and after treatment of internal hordeolum. Results. A total of 2127 operational taxonomic units were obtained from the two groups of samples, and there was no significant difference in alpha diversity before and after eyelid cleaning. At the phylum level, there was no significant difference between the two groups. The predominant phyla in Group A included the following: Firmicutes ( 32.78 % ± 20.16 % ), Proteobacteria ( 26.73 % ± 7.49 % ), Acidobacteria ( 10.58 % ± 11.45 % ), Bacteroidetes ( 9.05 % ± 6.63 % ), Actinobacteria (8.48% ±1.77%), and Chloroflexi ( 3.15 % ± 3.12 % ), while those in Group B were the following: Proteobacteria ( 31.86 % ± 9.69 % ), Firmicutes ( 29.07 % ± 24.20 % ), Acidobacteria ( 11.33 % ± 7.53 % ), Actinobacteria ( 7.10 % ± 1.98 % ), Bacteroidetes ( 5.39 % ± 5.17 % ), and Chloroflexi ( 3.89 % ± 3.67 % ). Starting from the class level, significant differences in microbial communities were detected before and after eyelid cleaning ( P < 0.05 ). Linear discriminant analysis effect size analysis showed the core flora in Group A microbiome comprising Actinobacteria, Staphylococcus, Staphylococcaceae, Staphylococcus aureus, Ruminococcacea UCg-014, Ruminococcacea-UCG-014, Halomonadaceae, Neisseria, Methylobacterium, Frankiales, and Neisseria sicca, while those in Group B microbial were Streptococcus sp., Blautia, Bifidobacterium pseudocatenulatum, Subdoligranulum, Subdoligranulum variabile, Faecalibacterium, and Faecalibacterium prausnitzii. Conclusion. Eyelid cleaning with hypochlorous acid eyelid wipes does not change the biodiversity in the meibomian gland secretions of patients with internal hordeolum. Hypochlorous acid eyelid wipes may affect the internal hordeolum through broad-spectrum antibacterial action to effectively reduce the relative abundance of symbiotic pathogens, such as Staphylococcus, Neisseria, Actinomycetes, and Ruminococcus and increase that of Faecalibacterium prausnitzii and other symbiotic probiotics with anti-inflammatory effects.
Background Alveolar echinococcosis (AE) is a severe parasitic zoonosis, caused by the larval stage of Echinococcus multilocularis. Identification of the antigens eliciting acquired immunity during infection is important for vaccine development against Echinococcus infection. Methods First of all, we predicted the physical and chemical properties and protein structure characteristics of E. multilocularis calreticulin (EmCRT), and then constructed the recombinant plasmid pET28a-EmCRT. Secondly, the constructed recombinant plasmid was transformed into E. coli BL21 and induced to produce recombinant EmCRT (rEmCRT). Then the antigen specificity and the calcium-binding property of rEmCRT was detected. Thirdly, the expression level and distribution of Emcrt gene in PSCs and vesicles were analysed. And the protective immunity induced by rEmCRT immunized mice and the corresponding mechanismwas investigated in the end. Results We identified that EmCRT, a ubiquitous protein with Ca2+-binding ability, elicited protective immunity against E. multilocularis infection. EmCRT was found to be expressed on the surface of E. multilocularis larvae as well as in the secreted products of metacestode vesicles and protoscoleces (PSCs). Recombinant EmCRT formulated with Freund’s adjuvant (FA) elicited both IgG1 and IgG2a antibodies, with a higher IgG1/IgG2a ratio indicating a predominant Th2 response in vaccinated mice, and stimulated mouse lymphocytes from the spleen to produce high levels of Th1 (IFN-γ, IL-2) and Th2 (IL-4, 5, 10) cytokines. The protection was around 43.7% after challenged with E. multilocularis PSCs. Conclusions Our results suggest that EmCRT is an immunodominant protein secreted by E. multilocularis larvae and contributes to the induction of partial protective immunity in vaccinated mice against Echinococcus infection. Therefore, EmCRT could be a novel and potential candidate vaccine against AE.
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