A Dendritic Cells-Targeting Nano-Vaccine by Coupling Polylactic-Co-Glycolic Acid-Encapsulated Allergen with Mannan Induces Regulatory T Cells.

BACKGROUND: The efficacy of allergen-specific immunotherapy (AIT) is mainly depended on the tolerogenic immune responses elicited. Properly conjugated nano-vaccine has the advantages of both specific targeting and continuous and on-demand release of allergen. OBJECTIVES: The aim of this study is to investigate the effects of a dendritic cells (DCs)-targeting nano-vaccine for AIT. METHODS: The nano-vaccine was produced by coupling polylactic-co-glycolic acid (PLGA)-encapsulated ovalbumin (OVA) with mannan. Allergen capture, human monocytes-derived DCs (hMoDCs) activation, and T cells responses were assessed by flow cytometry, confocal microscopy, quantitative real-time PCR, ELISA, and Cytometric Bead Array. Balb/c mice were immunized with the nano-vaccines, and the immune responses were analyzed. RESULTS: OVA-PLGA nanoparticle (NP) displayed favorable safety profile. OVA-mannan-PLGA NP was captured more efficiently by hMoDCs than OVA-PLGA NP, which was mediated mainly through DC-specific intercellular adhesion molecule 3-grabbing nonintegrin. A tolerogenic phenotype of hMoDCs was induced by OVA-mannan-PLGA NP, but not OVA-PLGA NP, and increased number of regulatory T (Treg) cells was generated subsequently in in vitro coculture. Immunization of Balb/c mice with OVA-mannan-PLGA NP resulted in lower serum level of OVA-specific immunoglobulins and less production of pro-inflammatory cytokines in splenocytes culture than the mice immunized with OVA-PLAG NP, PLGA NP, or OVA, while the number of splenic Treg cells was higher in OVA-mannan-PLGA group than in other groups. Moreover, preimmunization with OVA-mannan-PLGA NP significantly inhibited the Th2 immune response induced by OVA sensitization. CONCLUSIONS: The biocompatible PLGA-encapsulated OVA coupling with mannan has augmented ability for tolerance induction and could be developed as a novel vaccine for AIT.

The Development and Initial Validation of PUMC Localized Scleroderma Facial Aesthetic Index: A Pilot Study.

BACKGROUND: Localized scleroderma (LoS) is an autoimmune connective tissue disorder leading to serious long-term aesthetic impairment on patients. Objective evaluation methods are badly needed to facilitate the evaluation of the surgical treatment on individual patients and clinical studies. OBJECTIVE: To develop and assess the reliability and validity of Peking Union Medical College LoS facial aesthetic index (PUMC LoSFAI). METHODS: Twelve experts devoted their time and resources in the development and validation. LoS patients in the stable phase were recruited. Reliability and validity was then assessed. LoS patients were evaluated by two plastic surgeons using PUMC LoSFAI and LoS skin damage index (LoSDI). The PUMC LoSFAI comprises 4 domains for the local assessment (surface area of lesion, dyspigmentation, skin thickness and soft tissue atrophy) and 3 domains for the overall assessment (facial symmetry, proportion and profile) to describe LoS facial aesthetic impairment. Face-Q was completed by patients at each visit. RESULTS: Thirty-two LoS patients had 96 visits, during which 138 lesions were assessed. PUMC LoSFAI and 7 domains demonstrated substantial to excellent inter- and intra-rater reliability (ICC 0.995, κw 0.72-0.91, r 0.85-0.99, respectively). Seven domains considered to be important to extremely important variables (mean rank 3.2-3.8) had high I-CVI (> 0.78) and S-CVI (0.93). PUMC LoSFAI correlated excellently with LoSDI (r = 0.933, P < 0.001), and correlated fairly with Face-Q (r = - 0.399, P = 0.001). CONCLUSIONS: PUMC LoSFAI was developed and evaluated to play as a tool of aesthetic impairment assessment for LoS patients, which may facilitate the evaluation of the treatment on individual patients and clinical studies. PUMC LoSFAI demonstrated high reliability and validity, and further study in larger patient samples is needed to confirm these preliminary findings. LEVEL OF EVIDENCE IV: This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine Ratings, please refer to Table of Contents or online Instructions to Authors www.springer.com/00266 .

Dermatopathological features and successful treatment with topical antioxidant for ichthyosiform lesions in Mitchell syndrome caused by an ACOX1 variant.

Peroxisomal acyl-CoA oxidase 1 (ACOX1), is a peroxisomal enzyme that catalyzes ß-oxidation of very-long-chain fatty acids (VLCFA). The gain-of-function variant p.Asn237Ser in ACOX1 has been shown to cause Mitchell syndrome (MITCH), a neurodegenerative disorder characterized by episodic demyelination, hearing loss, and polyneuropathy, through the overproduction of hydrogen peroxide. Only eight cases of MITCH have been reported. While all these patients experienced cutaneous abnormalities, detailed skin features and potential treatment have not been documented. Herein, we report two MITCH patients who harbored a de novo heterozygous variant p.Asn237Ser in ACOX1 and experienced progressive ichthyosiform erythroderma. Skin histopathology revealed hyperkeratosis and parakeratosis with focal hypogranulosis as well as dyskeratotic keratinocytes. Lipid accumulation in the epidermis was observed using Oil Red O staining. Both patients exhibited a remarkable response to treatment with the topical antioxidant N-acetylcysteine (NAC), with Patient 1 achieving complete recovery after 3 months of consistent treatment. This study provides the first comprehensive description of the clinicopathological characteristics and effective treatment of skin lesions in MITCH patients. The successful treatment with topical NAC suggests excessive reactive oxygen species might play a significant role in the pathogenesis of skin lesions in MITCH.

Mendelian randomization study and mediation analysis about the relation of inflammatory bowel disease and diabetic retinopathy: the further exploration of gut-retina axis.

Background: The concept of the gut-retinal axis proposed by previous scholars primarily focused on the relationship between intestinal microbiota and retinal diseases, and few further expanded the relationship between intestinal diseases and retinal diseases. To further substantiate the concept of the gut-retinal axis, we analyzed inflammatory bowel disease (IBD) and diabetic retinopathy (DR) using Mendelian randomization (MR), and use mediation analysis to further explore the potential substances that influence this causal relationship. Methods: The genome-wide association study's (GWAS) summary statistics for genetic variations were utilized in a Mendelian randomization (MR) investigation. GWAS data on IBD (including ulcerative colitis (UC), Crohn's disease (CD), and IBD) for non-Finnish Europeans (NFE) were sourced from published articles. In contrast, data on DR (including DR and diabetic maculopathy (DMP)) were obtained from FinnGen R9. The causal relationship has been investigated using inverse variance weighted (IVW), MR-Egger, and weighted median and sensitivity analysis was applied to verify the stability of the results. In addition, we applied mediation analysis to investigate whether circulating inflammatory proteins and plasma lipids played a mediating role, and calculated its effect ratio. Results: The causal relationship between IBD and DR was discovered by employing the inverse variance weighted (IVW) method and weighted median method. In forward MR, UC was significantly associated with lower risk of DR (IVW: OR=0.874; 95%CI= 0.835-0.916; P value= 1.28E-08) (Weighted median: OR=0.893; 95%CI= 0.837-0.954; P value= 7.40E-04). In reverse MR, it was shown that DR (IVW: OR=0.870; 95%CI= 0.828-0.914; P value= 2.79E-08)(Weighted median: OR=0.857; 95%CI= 0.801-0.916; P value= 6.40E-06) and DMP (IVW: OR=0.900; 95%CI= 0.865-0.937; P value= 3.34E-07)(Weighted median: OR=0.882; 95%CI= 0.841-0.924; P value= 1.82E-07) could reduce the risk of CD. What's more, DR is associated with a lower risk of IBD according to genetic prediction (IVW: OR=0.922; 95%CI= 0.873-0.972; P value= 0.002) (Weighted median: OR=0.924; 95%CI= 0.861-0.992; P value= 0.029). Fibroblast growth factor 21 (FGF21), phosphatidylcholine (PC), and triacylglycerol (TG) serve as mediators in these relationships. Conclusions: Our research offers novel insights and sources for investigating the gut-retina axis in the genetic relationship between IBD and DR. We discover four mediators and more about the association between the intestine and retinal disorders and provide more evidence for the gut-retinal axis theory.

IL-4-induced decrease in both the number and CTLA-4 expression of Treg impairs suppression of Th2 type inflammation in severe atopic dermatitis.

BACKGROUND: Treg plays a pivotal role in the suppression of Th2 cell and the maintenance of immune homeostasis. The precise molecular mechanism underlying the disruption of Treg suppression of Th2 cell and the promotion of Th2 type inflammation in allergic diseases remains elusive. OBJECTIVE: This study aims to investigate the molecular mechanism underlying quantitative and functional changes of Treg in AD. METHODS: The molecular mechanism was investigated using flow cytometry, mRNA sequencing, co-culture experiments, co-immunoprecipitation, chromatin immunoprecipitation, and bisulfite sequencing in vitro or in AD mice model and patients with AD. RESULTS: Increased proportion of Treg was detected in mild and moderate AD. Conversely, characteristic decrease in both the number and CTLA-4 expression of Treg was relevant to serum IL-4 level in severe AD patients, which was verified under a high concentration of IL-4 treatment in vitro. The underlying mechanism is that IL-4/pSTAT6 pathway recruits DNMT1 and HDAC2 to inhibit transcriptional regulation of Foxp3 and CTLA-4 loci. High level of IL-4 impaired the suppression of Treg against Th2 cell differentiation mediated by CTLA-4, and blockade of IL-4Rα signaling in Treg restored Treg number and suppression of Th2 cell in AD model mice and patients with AD. CONCLUSION: The number of Treg is relevant to stratification of severity and serum IL-4 level in patients with AD. Abnormal high level of IL-4 epigenetically triggers a decrease in both the number and CTLA-4 expression of Treg. The reduced expression of CTLA-4 on Treg induced by IL-4 impairs suppression of Th2 cell differentiation.

Lymphatic microvessel density as a predictive marker for the recurrence time of pterygium: a three-year follow-up study.

PURPOSE: To investigate whether lymphatic microvessel density (LMVD) could be used as a predictive marker for the recurrence time of pterygia. METHODS: This was a prospective case series study. Ninety-six patients with unilateral eye primary nasal pterygia were included. The patients were clinically evaluated to grade the severity of their pterygia (32 were Grade 1, 29 were Grade 2, and 35 were Grade 3) before they underwent bare sclera resection with the use of mitomycin C. Excised tissues from the 96 patients and the ten normal nasal conjunctiva obtained from age-matched donor eyeballs (controls) were immunostained with LYVE-1 and CD31 monoclonal antibodies to evaluate LMVD and blood microvessel density (BMVD). The patients were followed up for three years or until pterygium recurrence was identified, which was defined as fibrovascular regrowth past the limbus in a previously compromised area. The recurrence time (RT) for a pterygium was calculated, and its relationship with LMVD and/or BMVD was statistically analyzed. RESULTS: In total, there were 24 cases of pterygium recurrence. The recurrence rate of Grade 1 was 28.1% (9/32), Grade 2 was 24.1% (7/29), and Grade 3 was 22.9% (8/35), as classified in the primary pterygium (p>0.05); the overall recurrence rate was 25% (24/96) for all patients during the three-year follow-up. In the tissue analysis, there were a small number of CD31 (+), LYVE-1(-) BMVD and only a few CD31 (weak), LYVE-1(+) LMVD in the ten normal nasal conjunctiva tissues. BMVD and LMVD increased significantly in the pterygium tissue compared to the control tissue and were significantly correlated with the width and area of pterygium in Grades 1-3 (all p values <0.05). RT was not correlated with BMVD or pterygium grade, but LMVD was significantly and negatively correlated with RT within each group and in the total patient cohort. Furthermore, we determined that an LMVD greater than 20 in the surgical specimens predicted pterygium recurrence. CONCLUSIONS: The LMVD of surgical specimens is an independent risk factor and a valuable predictive factor for the recurrence time of pterygia.

Effects of chemokine receptor CCR7 in the pathophysiology and clinical features of the immuno-inflammatory response in primary pterygium.

OBJECTIVE: Chemokine receptor 7 (CCR7) has been considered a critical biomarker in inflammation and the immune response; however, little is known about CCR7 in pterygia. This study aimed to investigate whether CCR7 participates in the pathogenesis of primary pterygia and how CCR7 affects the progression of pterygia. METHODS: This was an experimental study. Slip-lamp photographs of 85 pterygium patients were used to measure the width, extent, and area of pterygia with computer software. Pterygium blood vessels and general ocular redness were quantitatively analyzed with a specific algorithm. The expression of CCR7 and its ligands C-C motif ligand 19 (CCL19) and C-C motif ligand 21 (CCL21) in control conjunctivae and excised pterygia collected during surgery were analyzed by quantitative real-time polymerase chain reaction (qRT-PCR) and immunofluorescence staining. The phenotype of CCR7-expressing cells was identified by costaining for major histocompatibility complex II (MHC II), CD11b or CD11c. RESULTS: The CCR7 level was significantly increased by 9.6-fold in pterygia compared with control conjunctivae (p = 0.008). The higher the expression of CCR7 was, the more blood vessels appeared in pterygia (r = 0.437, p = 0.002) and the more general ocular redness was (r = 0.51, p < 0.001) in pterygium patients. CCR7 was significantly associated with pterygium extent (r = 0.286, p = 0.048). In addition, we found that CCR7 colocalized with CD11b, CD11c or MHC II in dendritic cells, and immunofluorescence staining showed that CCR7-CCL21 is a potential chemokine axis in pterygium. CONCLUSIONS: This work verified that CCR7 impacts the extent of primary pterygia invading the cornea and inflammation at the ocular surface, which may provide a possibility for a further in-depth understanding of the immunological mechanism in pterygia.

MSCohi-O lenses for long-term retention of mesenchymal stem cells on ocular surface as a therapeutic approach for chronic ocular graft-versus-host disease.

Chronic ocular graft-versus-host disease (oGVHD) is a common complication of allogeneic hematopoietic stem cell transplantation (allo-HSCT) and can lead to vision loss if not diagnosed and treated promptly. Currently, no approved drugs exist for oGVHD treatment. However, umbilical cord-derived mesenchymal stem cells (UCMSCs) have known immunoregulatory properties and have been employed in clinical trials for immune-mediated diseases. To address oGVHD, the application of UCMSCs to the ocular surface is a logical approach. Intravenous administration of UCMSCs poses risks, necessitating topical and local delivery. Retaining UCMSCs on the ocular surface remains a challenge. To overcome this, we invented mesenchymal stem cell-coating high oxygen-permeable hydrogel lenses combining UCMSCs and machinery to enable the long-term retention of UCMSCs on the ocular surface. Animal model experiments demonstrated that these lenses effectively retained UCMSCs, providing therapeutic benefits by decreasing corneal inflammation and damage, and inhibiting immune rejection and response, all crucial aspects in oGVHD treatment.

Attenuation of allergen-specific immunotherapy for atopic dermatitis by ectopic colonization of Brevundimonas vesicularis in the intestine.

Allergen-specific immunotherapy (AIT) has shown beneficial effects against atopic dermatitis (AD); however, the mechanisms and parameters underlying the efficacy of AIT remain unclear. Here, we report that the community structure and function of the oral and gut microbiota are changed in patients with AD undergoing AIT. Transplantation of fecal microbiota from patients who respond well to AIT improves AD-like dermatitis in mice. The abundance of Brevundimonas vesicularis in the gut of AD patients has been found to be positively correlated with disease severity and is decreased following AIT. Furthermore, we find that B. vesicularis from the oral cavity might ectopically colonize the gut of AD patients. In AD model mice, meanwhile, B. vesicularis promotes the skewing of the Treg/Th17 balance toward Th17 polarization and attenuates the efficacy of ovalbumin-specific immunotherapy. Our findings provide potential strategies for the optimization of AIT for AD via the modulation of the gut microbiota.

[Dynamic aspects of corneal lymphatic vessels after keratoplasty in rats].

OBJECTIVE: To study the development of lymphatic vessels after keratoplasty and to explore the molecular mechanisms of corneal lymphangiogenesis in transplanted corneas. METHODS: Experimental research. The development of corneal lymphangiogenesis was examined by LYVE-1 immunohistochemistry and whole mount immunofluorescence 1, 3, 7, 10, 14, 30 and 60 days after corneal transplantation, then lymphatic vessels counting (LVC)was evaluated. The expression of vascular endothelial growth factor-C (VEGF-C) in transplanted corneas was examined by immunohistochemistry, enzyme-linked immunosorbent assay (ELISA), and real time-PCR at same time. In addition, the inflammatory index (IF) was recorded at each time point. The association of VEGF-C and IF with LVC in transplanted corneas was examined. Analysis of the significance of differences between two groups was performed using paired Student's t-test. Pearson's analysis was used to analyze the correlation between VEGF-C, IF and LVC. RESULTS: Corneal lymphangiogenesis occurred in the stroma with LVC (1.8 ± 0.3) on Day 3, then developed and reached the peak with LVC (9.1 ± 1.5) on Day 14 after corneal transplantation. Both VEGF-C protein and mRNA up-regulated dramatically in rat transplanted corneas. The immunoreactivity reached the peak on the 3(rd) day and the 14(th) day after keratoplasty. Compared with the expression of VEGF-C mRNA (1.62 ± 0.08 copies/g) on Day 3, the expression of VEGF-C mRNA (2.48 ± 0.03 copies/g) significantly increased 14 days after the transplantation (t = 4.296, P = 0.02). LVC was strongly and positively correlated with IF (r = 0.55, P = 0.003) and the expression of VEGF-C mRNA (r = 0.51, P = 0.003). CONCLUSIONS: Corneal lymphangiogenesis correlates closely with corneal inflammation. The increased expression of VEGF-C in the cornea may be one of the important molecular mechanisms in the occurrence of corneal lymphangiogenesis after keratoplasty.

Transglutaminase 3 Attenuates Skin Inflammation in Psoriasis by Inhibiting NF-κB Activation through Phosphorylated STAT3-TET3 Signaling.

Transglutaminase 3 (TGM3) protects against skin inflammation in psoriasis, but the precise role and mechanism of action of TGM3 in the pathogenesis of psoriasis remain unclear. In this study, we show that TGM3 expression was increased in the skin lesions of patients with psoriasis and a mouse model of imiquimod-induced psoriatic dermatitis. TGM3 overexpression decreased the production of proinflammatory factors in cultured primary keratinocytes stimulated with psoriasis-related cytokines. TGM3 inhibited the phosphorylation of signal transducer and activator of transcription 3 and the recruitment of ten-eleven translocation 3 to the p65 gene promoter, resulting in decreased promoter demethylation and subsequent suppression of proinflammatory cytokine/chemokine production. TGM3-induced inhibition of phosphorylated p65 might also decrease ten-eleven translocation 3 expression. Moreover, topical application of Tgm3-specific small interfering RNA or the pan-transglutaminase inhibitor cysteamine exacerbated skin inflammation in mice with imiquimod-induced psoriatic dermatitis. Our study revealed an epigenetic pathway mediated by the interaction between TGM3 and ten-eleven translocation 3 in keratinocytes for regulation of skin inflammation in psoriasis, providing a potential target for psoriasis treatment.

Clinicopathological correlation analysis of (lymph) angiogenesis and corneal graft rejection.

PURPOSE: To investigate the relationship of (lymph) angiogenesis and survival time of human cornea grafts. METHODS: This was a case series study. A total of 250 patients requiring a second keratoplasty were screened according to our inclusion criteria: 1) only cases with primary non-inflamed cornea diseases were included; and 2) all primary diseased cornea specimens from the first corneal transplantation were confirmed not to have hemangiogenesis or lymphangiogenesis. The included patients were analyzed retrospectively and followed up for the survival time of the first and second grafts. Blood vessel content (BVC) and lymphatic vessel content (LVC) were assessed in the primary diseased cornea; first rejected grafts (including BVC(1) and LVC(1)) and second rejected grafts (including BVC(2) and LVC(2)) were assessed by immunohistochemistry. The survival times of the first (STG(1)) and second (STG(2)) rejected corneal grafts were calculated and the relationship between human corneal (lymph) angiogenesis and STG was statistically analyzed. RESULTS: After screening, only 23 patients (23 eyes) were included. Their primary cornea diseases were non-inflamed, including keratoconus (n=14), leukoma (n=5), and Fuchs endothelial dystrophy (n=4). The mean duration of follow up was 36 months after the second keratoplasty. In all, 55 cornea specimens from different times following penetrating keratoplasty were collected and examined, including 23 primary non-inflamed corneas (without angiogenesis), 23 first rejected corneal grafts (all with hemangiogenesis, but only six cases with blown lymphatic vessels), and nine rejected corneal grafts (including six cases identified with lymphangiogenesis in the first rejection, all with lymphangiogenesis and hemangiogenesis). Based on our statistical analysis, STG(1) was correlated with LVC(1) but not with BVC(1) or (LVC(1)+BVC(1)), while STG(2) was correlated with (LVC(1)+LVC(2)), LVC(1), LVC(2), (LVC(2)+BVC(2)) and (LVC(1)+BVC(1)) but not with BVC(1) or BVC(2). CONCLUSIONS: The survival time of human cornea grafts is related to both lymphangiogenesis and hemangiogenesis. Lymphangiogenesis only occurred in some rejected cases, but it seems to be a signal of poor prognosis for the new allograft.

Preventive scleral buckling and silicone oil tamponade are important for posttraumatic endophthalmitis successfully managed with vitrectomy.

AIMS: Posttraumatic endophthalmitis (PTE) managed with pars plana vitrectomy (PPV) and a variety of other methods often resulted in retinal detachment (RD), which deteriorates postoperative visual acuity (VA). We investigated initial ocular conditions, surgical management and outcomes of PTE patients and analyzed their relationship in order to find the necessary management for different patients' conditions. METHODS: Retrospective study. Medical records of 62 consecutive patients who were diagnosed with PTE and managed with PPV were retrospectively analyzed. The included patients were divided into two groups depending on their initial VA, with light perception (LP) as borderline: group A (12 cases; VA=LP) and group B (50 cases; VA>LP). Specific variables related to surgical outcomes were analyzed, including: initial VA; intraocular foreign body; location and length of the wound; microbiologic test; cataract extraction; scleral buckling; silicon oil tamponade; gas tamponade, and retinal restoration. Data were statistically analyzed by the χ2 test, by univariate analysis for predictors and by multivariate logistic regression analysis. RESULTS: RD happened in 19 (30.6%) of the 62 cases included. There were 6 cases (50%) in group A and 13 cases (26%) in group B which resulted in RD. Statistical analysis demonstrated that none of the related factors were correlated to the occurrence of RD (all factors: p>0.05) in group A, but in group B, scleral buckling (with: 2/22, 9%; without: 11/28, 39%; p<0.05) and silicone oil tamponade (with: 0/6; without: 13/44, 30%; p<0.05) were statistically significantly correlated to the occurrence of RD. CONCLUSIONS: Preventive scleral buckling and silicone oil tamponade are important for patients with good initial VA (VA>LP) to be successfully managed by PPV.

The Important Role of the Chemokine Axis CCR7-CCL19 and CCR7-CCL21 in the Pathophysiology of the Immuno-inflammatory Response in Dry Eye Disease.

Purpose: To explore whether CCR7-CCL19 and CCR7-CCL21 affect the pathophysiology of the dry eye disease (DED) immuno-inflammatory response using a murine model.Methods: The mRNA expression levels of CCR7, CCL19, CCL21 and VEGF-C within corneas in DED mice were detected by real-time PCR. Immunofluorescence and flow cytometric analyses were performed to mark dendritic cells (DCs) and detect correlations among CCR7, CCL19, CCL21 and lymphatic vessels.Results: CCR7, CCL19 and CCL21 expression was dramatically increased during the development of DED. In addition, CCR7, which is expressed in DCs, was located inside and around lymphatic vessels and colocalized with CCL19 or CCL21. Positive correlations were observed between CCR7 and CCL19 (P < .01, r = 0.862), CCL21 (P < .01, r = 0.759), and VEGF-C (P < .05, r = 0.607).Conclusions: Our study revealed that both the CCR7-CCL19 and CCR7-CCL21 chemokine axis are important for DC migration to lymphatic vessels, but CCL19 may have a greater effect on DED than CCL21.

Treatment With Melatonin After Corneal Graft Attenuates Rejection.

Background: Immunologic graft rejection is the main complication of corneal transplants. This study aimed to investigate the effect of melatonin (MT) on the rejection of corneal transplantation. Methods: Corneal allografts were performed by grafting corneas from BALB/C mice to C57BL/6 hosts. MT (50 mg/kg) was intraperitoneally injected into the hosts every day from the day of transplantation. The survival of grafts was observed by slit lamp biomicroscopy, and inflammatory cell infiltration was detected by hematoxylin and eosin staining and immunohistochemistry. The balance of Teff and Treg immune cells in draining lymph nodes (DLNs) was detected by flow cytometry. The levels of cytokines related to the grafts and DLNs were detected using real-time fluorescence quantitative PCR. Additionally, we used the mouse macrophage line RAW264.7 to study the effect of MT on the activation of NLRP3 inflammatory body. Results: MT treatment improved the graft survival rate, reduced inflammatory cell infiltration in the graft, decreased the percentage of Th1/Th17 cells in the DLNs, and increased the percentage of Treg cells. Melatonin inhibited the activation of the NLRP3 inflammasome, thereby reducing the expression of IL-1ß and other related proinflammatory cytokines such as MCP-1, MIP-1, NLRP3, ASC, TNF-a and VEGF-A (all p < 0.05). Conclusion: Our study demonstrates that MT promotes the survival of mouse corneal grafts by inhibiting NLRP3-mediated immune regulation, reducing immune cell activation and cell migration, and inhibiting the production of inflammatory-related cytokines. Treatment with MT might provide a potential clinical therapeutic target for corneal transplantation.

Characterization of an i.p. D-galactose-induced cataract model in rats.

Cataracts have been identified as a main cause of global visual impairment and blindness; in addition, diabetic and aging cataracts are the most common types. The aim of this project was to develop a suitable animal model and investigate the key points of the mechanisms by which intraperitoneal (i.p.) injection of D-galactose forms cataracts. We optimized a method to investigate the safest and effective method and dosage; rats in Group H were treated with 50% D-galactose 15 g/kg i.p. twice daily based on the 11 different treatment methods. The simple oral group showed considerable differences in the same observed time, while the i.p. group showed relatively uniform cataracts due to intake of the same dose of D-galactose. The data suggest that i.p. injection of galactose is a relatively more successful and stable cataract-inducing method with a low mortality rate. Based on this model, we found that Na+/K+ ratios had important relevance for galactose cataract formation, and we used scanning electron microscopy (SEM), inductively coupled plasma mass spectrometry (ICP-MS), enzyme-linked immunosorbent assay (ELISA) and immunofluorescence examinations to test and verify this.

Study time, glasses utilization and age affect quality of life among senior first-year Chinese myopia students.

PURPOSE: To compare the quality of life of senior first-year students with normal vision and myopia, and to explore the risk factors related to quality of life in students with myopia. METHODS: In this study, 1103 senior first-year students were enrolled in ten high schools. These students were divided according to the diopter degree, with 916 myopia students and 187 normal vision students. Visual function indexes, such as naked eye vision, were measured and recorded, and social demographic indexes and the National Eye Institute 25-Item Visual Function Questionnaire (NEI VFQ-25) was used. The differences in quality of life between the two groups were compared. Multiple linear regression analysis was used to explore the possible risk factors for quality of life in myopia students. RESULTS: In the NEI VFQ-25, the total quality of life scores of myopia students (77.06 ± 15.66) were lower than those of normal vision students (85.49 ± 12.37). The difference was statistically significant (p = 0.007). In the correlation analysis, the total scores of quality of life in myopia students were positively correlated with wearing glasses (p = 0.049), and were negatively correlated with study time (p = 0.029). Multiple linear regression analysis showed that study time, wearing glasses and age were risk factors affecting quality of life in myopia students. CONCLUSION: Our results show that senior first-year myopia students have lower quality of life scores than students with normal vision. Study time, wearing glasses and age are risk factors for quality of life in senior first-year myopia students.

A New Formulation of Probiotics Attenuates Calcipotriol-Induced Dermatitis by Inducing Regulatory Dendritic Cells.

Atopic dermatitis (AD) is a recurrent chronic inflammatory skin disease affecting up to 30% of the children population, and immuno-regulatory therapy that could modify the course of disease is urgently needed. Probiotics have demonstrated therapeutic effects on AD and could potentially regulate the disease process. However, the efficacy of probiotics for AD is inconsistent among different studies, which is mainly due to the elusive mechanism and different species and (or) strains used. In this study, we designed a mixture of five strains of probiotics (named IW5) and analyzed the effect and mechanism of IW5 on calcipotriol (MC903)-induced AD-like dermatitis. We found that IW5 significantly alleviated skin inflammation of the MC903-induced AD in mice. Administration with IW5 induced increased production of regulatory T cells and regulatory dendritic cells (DCregs) in the mesenteric lymph nodes. We also found that the diversity of the gut microbiota in the mice with MC903-induced dermatitis was increased after IW5 administration, and the level of butyrate in the gut was elevated. In cell culture, butyrate induced the production of DCregs. Our study revealed the therapeutic effects of a newly designed probiotics mixture and uncovered a possible mechanism, providing a foundation for future clinical studies.