Attenuated airways inflammation and remodeling in IL-37a and IL-37b transgenic mice with an ovalbumin-induced chronic asthma.

BACKGROUND: Asthma is a common chronic respiratory disease characterized by airways inflammation, hyperresponsiveness and remodeling. IL-37, an anti-inflammatory cytokine, consists of five splice isoforms, that is, a-e. Although it has been previously shown that recombinant human IL-37b is able to inhibit airway inflammation and hyperresponsiveness in animal models of asthma, the effects and difference of other IL-37 isoforms, such as IL-37a on features of asthma are unknown. METHODS: Animal models of chronic asthma were established using IL-37a and IL-37b transgenic mice with C57BL/6J background and wild-type (WT) mice sensitized and nasally challenged with ovalbumin (OVA). Airway hyperresponsiveness was measured using FlexiVent apparatus, while histological and immunohistological stainings were employed to measure airways inflammation and remodeling indexes, including goblet cell metaplasia, mucus production, deposition of collagen, hypertrophy of airway smooth muscles and pulmonary angiogenesis. RESULTS: Compared to WT mice, both IL-37a and IL-37b transgenic mice had significant reduced airway hyperresponsiveness and the declined total numbers of inflammatory cells, predominant eosinophils into airways and lung tissues. Furthermore, all features of airways remodeling, including degrees of mucus expression, collagen deposition, hypertrophy of smooth muscles, thickness of airways and neovascularization markedly decreased in IL-37 transgenic mice compared with OVA-treated WT mice. CONCLUSION: Our data suggest that both IL-37a and IL-37b isoforms are able to not only ameliorate airways inflammation and airways hyperresponsiveness, but also greatly reduce airways structural changes of animal models of chronic asthma.

New keratoconus staging system based on OCT.

PURPOSE: To establish a numerical spectral-domain optical coherence tomography (SD-OCT)-based keratoconus (KC) staging system and compare it with existing KC staging systems. SETTING: Eye Hospital of Wenzhou Medical University, Wenzhou, China. DESIGNS: Retrospective case-control study. METHODS: Scheimpflug tomography, air-puff tonometry, and SD-OCT were performed on 236 normal and 331 KC eyes. All SD-OCT-derived parameters of the corneal epithelium and stroma were evaluated based on their receiver operating characteristic (ROC) curves, area under the curve (AUC), sensitivity, and specificity to discriminate between normal and KC eyes. The best performing parameters were subsequently used to create an OCT-based staging system, which was compared with existing tomographic and biomechanical staging systems. RESULTS: 236 eyes from 236 normal patients and 331 eyes from 331 KC patients of different stages were included. The highest ranked AUC ROC SD-OCT parameters, derived from stroma and epithelium, were stroma overall minimum thickness (ST: AUC 0.836, sensitivity 90%, specificity 67%) and epithelium overall SD (EP: AUC 0.835, sensitivity 75%, specificity 78%). A numerical SD-OCT staging system called STEP including 2 parameters-"ST" and "EP"-with 5 stages was proposed. CONCLUSIONS: The new SD-OCT-based KC staging system is the first to take the epithelium with its sublayer stroma information into account, showing a strong agreement to the existing staging systems. This system could be incorporated into daily practice, potentially leading to an overall improvement in KC treatment and follow-up management.

CXCL8, CXCL9, CXCL10, and CXCL11 as biomarkers of liver injury caused by chronic hepatitis B.

Background: Chronic hepatitis B (CHB) remains a significant global health problem, leading to recurrent inflammation and liver-damaging diseases such as fibrosis, cirrhosis, and hepatocellular carcinoma (HCC). Currently, although diagnostic markers for CHB are well established, the indicators for predicting liver injury caused by hepatitis B virus (HBV) infection still need to be further explored. Thus, the identification of credible infectious indicators is urgently needed to facilitate timely clinical intervention and avoid the progression of disease malignancy. Methods: The Gene Expression Omnibus (GEO) database GSE83148 data set was used to explore the hub genes for HBV infection. The quantitative real-time polymerase chain reaction (qPCR) was used to identify the impact of HBV infection on the expression of hub gene at the cell level. At the same time, serum samples and clinical information were collected from healthy, HBV-free and CHB patients. The enzyme-linked immunosorbent assay (ELISA) was used to verify the results of cell experiments and Pearson correlation analysis was used to clarify hub genes correlation with HBV infection indicators and liver injury-related indicators. Finally, the Gene Expression Profiling Interactive Analysis (GEPIA) database was used to analyze the differences in the expression of hub gene in liver injury diseases. Results: Chemokine (C-X-C motif) ligand (CXCL)8, CXCL9, CXCL10, and CXCL11 were identified as hub genes in HBV infection. After HBV infection, the expression of the four chemokines was significantly increased and the concentrations secreted into serum were also increased. Moreover, the four chemokines were significantly correlated with HBV infection-related indicators and liver injury-related indicators, which were positively correlated with alanine aminotransferase (ALT), aspartate aminotransferase (AST) and hepatitis B e antigen (HBeAg), and negatively correlated with AST/ALT ratio and hepatitis B core antibody (HBcAb). In addition, the expression of CXCL9, CXCL10, and CXCL11 in HCC tissues was significantly higher than in normal tissues. Conclusion: Using a combination of bioinformatics, cell experiments, and clinical correlation analysis, this study showed that CXCL8, CXCL9, CXCL10, and CXCL11 can be used as serum biomarkers to forecast liver injury caused by HBV infection.

Combining Spectral-Domain OCT and Air-Puff Tonometry Analysis to Diagnose Keratoconus.

PURPOSE: To investigate the diagnostic capacity of spectral-domain optical coherence tomography (SD-OCT) combined with air-puff tonometry using artificial intelligence (AI) in differentiating between normal and keratoconic eyes. METHODS: Patients who had either undergone uneventful laser vision correction with at least 3 years of stable follow-up or those who had forme fruste keratoconus (FFKC), early keratoconus (EKC), or advanced keratoconus (AKC) were included. SD-OCT and biomechanical information from air-puff tonometry was divided into training and validation sets. AI models based on random forest or neural networks were trained to distinguish eyes with FFKC from normal eyes. Model accuracy was independently tested in eyes with FFKC and normal eyes. Receiver operating characteristic (ROC) curves were generated to determine area under the curve (AUC), sensitivity, and specificity values. RESULTS: A total of 223 normal eyes from 223 patients, 69 FFKC eyes from 69 patients, 72 EKC eyes from 72 patients, and 258 AKC eyes from 258 patients were included. The top AUC ROC values (normal eyes compared with AKC and EKC) were Pentacam Random Forest Index (AUC = 0.985 and 0.958), Tomographic and Biomechanical Index (AUC = 0.983 and 0.925), and Belin-Ambrósio Enhanced Ectasia Total Deviation Index (AUC = 0.981 and 0.922). When SD-OCT and air-puff tonometry data were combined, the random forest AI model provided the highest accuracy with 99% AUC for FFKC (75% sensitivity; 94.74% specificity). CONCLUSIONS: Currently, AI parameters accurately diagnose AKC and EKC, but have a limited ability to diagnose FFKC. AI-assisted diagnostic technology that uses both SD-OCT and air-puff tonometry may overcome this limitation, leading to improved treatment of patients with keratoconus. [J Refract Surg. 2022;38(6):374-380.].

Therapeutic Effects of Human Pluripotent Stem Cell-Derived Mesenchymal Stem Cells on a Murine Model of Acute Type-2-Dominated Airway Inflammation.

Allergic rhinitis and allergic asthma are the most common type-2 inflammatory diseases, which are hardly curable and cause heavy burden to general well-being. Mesenchymal stem cells (MSCs) are multipotent nonhematopoietic cells with potential immunomodulatory effects that have been showning to have a therapeutic effect on allergic diseases. Here, we investigated the effects of human induced pluripotent stem cell (iPSC)-derived MSCs on airway hyperresponsiveness and acute type-2-dominated inflammation throughout the upper and lower airways. In this study, human MSCs, MSC cell culture supernatant, and culture medium (control) was injected into the acute airway inflammatory model via the tail vein. Mouse behavioristics were recorded immediately and mouse lung function was measured 24 hours after the last ovalbumin (OVA) challenge. Histological staining, Luminex, Elisa and flow cytometry were employed to evaluate the effects on the production of total/OVA-specific IgG1 and IgE, cytokines expression in lung tissues, and inflammatory cells infiltration in the lung and spleen of the experimental mice. Expressions of eotaxin, IL-4, IL-5, IL-13, IL-33 in nasal and lung lavage were evaluated by Luminex and Elisa. We found that for this acute inflammatory mouse model, human MSC transplantation significantly mitigated the decreased motoring time and the increased lung function Rrs caused by OVA challenge. Serum OVA-IgG1, OVA-IgE, and eosinophil percentages in the splenocytes were significantly decreased. Injection of the MSC supernatant also showed the same trend, but not significantly changed. After treatment, IL-4 and IL-13 were significantly decreased in the lung tissue, and IL-5 and IL-13 were significantly decreased in lung lavage. In conclusion, both human MSC culture supernatant and cell transplantation could alleviate AHR and inflammation in acute inflammatory experimental animals, which demonstrated their potential for clinical therapeutics. Human iPSC-MSCs, MSC cell culture supernatant, or culture medium (control) was injected into the OVA-induced acute airway inflammatory model via the tail vein. Behavioral changes, AHR, serum OVA-specific IgG1 and IgE concentrations, and type-2 inflammations were alleviated.

Association of cigarette smoking with retinal capillary plexus: an optical coherence tomography angiography study.

PURPOSE: To investigate the association between cigarette smoking and retinal capillary plexus (RCP) using optical coherence tomography angiography (OCTA) and to examine whether potential vascular risk factors could impact their association. METHODS: This is a cross-sectional, community-based study. The Jidong Eye Cohort Study included participants aged ≥18 years in the Jidong community (Tangshan city, northern China) from August 2019 to January 2020. All participants underwent comprehensive ophthalmic examination and completed detailed smoking questionnaires. Retinal vessel density in the superficial and deep RCP was automatically measured using OCTA. RESULTS: Of the 2598 participants included in the study, 2026 (78.0%) never smoked and 572 (22.0%) had a history of smoking (494 [19.0%] current smokers and 78 [3.0%] former smokers). The median (interquartile range) age was 41 (34-52) years for the non-smoking group and 45 (35-54.5) years for the smoking group. Multivariable analysis showed that smoking history is associated with a low deep RCP vessel density in the parafovea (ß, -0.53; 95% confidence interval [CI], -0.82 to -0.24) and four quadrants. Increased smoking pack-years were associated with reduced deep RCP vessel density in the parafovea (p for trend <0.001) and four quadrants. The significant interaction between diabetes and smoking only was found for superficial RCP vessel density in the parafovea (p for interaction = 0.014) and four quadrants except for the temporal quadrants. CONCLUSIONS: Cigarette smoking is an independent risk factor for reduced deep RCP vessel density. Our findings imply the potential detrimental effect of smoking on the occurrence of ocular diseases.

Repeatability of Cornea and Sublayer Thickness Measurements Using Optical Coherence Tomography in Corneas of Anomalous Refractive Status.

PURPOSE: To assess the repeatability of epithelial, stromal, and total corneal thickness measurements with spectral-domain optical coherence tomography (SD-OCT; RTVue-XR; Optovue, Inc., Fremont, CA) in patients with myopia, keratoconus, and corneas after transepithelial photorefractive keratectomy (PRK), small incision lenticule extraction (SMILE), and femtosecond laser-assisted in situ keratomileusis (FS-LASIK). METHODS: A total of 352 eyes of 352 patients (75 myopic, 68 post-transepithelial PRK, 61 post-SMILE, 75 post-FS-LASIK, 20 mild keratoconus, and 53 advanced keratoconus eyes) were included. The epithelial, stromal, and total corneal thickness were recorded from the pachymetric map in the following four zones: (1) central 2-mm region, (2) eight paracentral regions within 2- to 5-mm diameter, (3) eight midperipheral regions within 5- to 7-mm diameter, and (4) eight peripheral regions within 7- to 9-mm diameter. Three successive scans were performed to evaluate the repeatability. RESULTS: For all zones up to 9-mm diameter, the coefficient of variation (CoV) for epithelial thickness measurements ranged from 1.7% to 3.5% for myopia, 2.6% to 6.2% for post-transepithelial PRK, 2.3% to 4.7% for post-SMILE, 4.0% to 6.3% for post-FS-LASIK, 2.5% to 6.2% for mild keratoconus, and 3.5% to 8.0% for advanced keratoconus. The CoV for stromal and total thickness measurements ranged from 0.2% to 2.0% for myopia, 0.7% to 4.2% for post-transepithelial PRK, 0.3% to 2.4% for post-SMILE, 0.3% to 1.9% for post-FS-LASIK, 0.6% to 3.0% for mild keratoconus, and 1.0% to 5.9% for advanced keratoconus. CONCLUSIONS: RTVue-XR SD-OCT showed excellent repeatability when generating the pachymetric map in myopic eyes. Measurements, especially for epithelial thickness, were relatively more variable for corneas with keratoconus and those having undergone keratorefractive surgery. [J Refract Surg. 2019;35(9):600-605.].

Effect of Dandelion Extracts on the Proliferation of Ovarian Granulosa Cells and Expression of Hormone Receptors.

BACKGROUND: In the current society, infertility related to age has become a social problem. The in vitro fertilization (IVF) success rate in women with poor ovarian response (POR) is very low. Dandelion extract T-1 (DE-T1) is an effective component of the extract from the leaves and stems of Taraxacum officinale, which is one of the medicines used in some patients with POR, but its molecular mechanism remains unclear. METHODS: Following IVF, ovarian granulosa cells (GCs) of sixty patients were extracted and divided into normal ovarian response (NOR) and POR groups. GCs were cultured in a dose-dependent and time-dependent manner with DE-T1, proliferation of GCs was determined by Cell Counting Kit-8 assay, and mRNA levels of insulin-like growth factor 1 receptor (IGF-1R), luteotropic hormone receptor (LHR), follicle-stimulating hormone receptor (FSHR), LHR, and CYP19A1 (aromatase) were determined by quantitative polymerase chain reaction. Progesterone and estradiol (E2) concentrations were determined by enzyme-linked immunosorbent assay. RESULTS: The cell viability gradually increased with the progressive increase in the DE-T1 concentration. Compared with the control group (without DE-T1), the mRNA expressions of FSHR, LHR, IGF-1R, and CYP19A1 were upregulated after the addition of DE-T1, especially in the 2.5% DE-T1 group (P < 0.01). The expression of IGF-1R was upregulated approximately 25 times (24.97 ± 4.02 times) in the POR group with 2.5% DE-T1. E2 and progesterone levels increased with the increasing DE-T1 concentration. There were highly significant differences in the E2 and progesterone secretion between the NOR and POR groups (P < 0.01). CONCLUSION: DE-T1 may promote steroid hormone synthesis by promoting GC proliferation and upregulating GC receptor expression, thereby improving ovarian endocrine function.

The Mechanism of Mitochondria-Mediated Pathway in the Apoptosis of Platelets in Immune-Induced Bone Marrow Failure.

Excessive platelet apoptosis is one of the pathogenic causes of immune-induced bone marrow failure (BMF). The aim of the present study was to explore the role of mitochondria-mediated pathway in the apoptosis of platelets in immune-induced BMF. An immune-induced BMF model was established in mice, which were randomly divided into three groups: normal control (CTL) group, BMF group and cyclosporine (CSA) group (n = 10 in each group). Mice were given 0.027 g/kg CSA daily in the CSA group. Platelet count (PLT), mitochondrial transmembrane potential (ΔΨm), cytochrome C (CytC), phosphatidylserine (PS), calcium ion (Ca²âº) and expression of proteins of the mitochondrial apoptotic pathway, including Bak, Bax, caspase-3, caspase-8 and caspase-9, was examined and compared. Compared with the CTL group, the BMF group had significantly a lower level of PLC and ΔΨm, but higher levels of CytC, PS, Ca²âº and higher expression levels of Bak, Bax, cleaved caspase-9 and cleaved caspase-3 (P < 0.05). CSA restored the above changes in the BMF model (P < 0.05). Further studies showed that intravenous injection of the caspase-9 inhibitor Z-LE(OMe)HD(OMe)-fluoromethylketone (FMK) into the mice could significantly inhibit apoptosis of the platelets and the effect of CSA treatment when compared to the BMF group, and exerted a better protective effect from apoptosis if the caspase-9 inhibitor was combined with the CSA treatment. These results revealed that platelet apoptosis may play an important role in the reduction of platelet of immune-induced BMF probably through the mitochondrial pathway.

Bioinspired Antioxidant Defense System Constructed by Antioxidants-Eluting Electrospun F127-Based Fibers.

Cells were continuously exposed to oxidative damage by overproduction of reactive oxygen species (ROS) when they contacted implanted biomaterials. The strategy to prevent cells from oxidative injures remains a challenge. Inspired by the antioxidant defense system of cells, we constructed a biocompatible and ROS-responsive architecture on the substrate of styrene-b-(ethylene-co-butylene)-b-styrene elastomer (SEBS). The strategy was based on fabrication of architectures through reactive electrospinning of mixture including SEBS, acylated Pluronic F127, copolymer of poly(ethylene glycol) diacrylate and 1,2-ethanedithiol (PEGDA-EDT), and antioxidants (AA-2G) and ROS-triggered release of AA-2G from microfibers to detoxify the excess ROS. We demonstrated that the stable and hydrophilic architecture was constructed by phase separation of SEBS/F127 components and cross-linking between polymer chains during electrospinning; the ROS-responsive fibers controlled the release of AA-2G and the interaction of AA-2G with ROS reduced the oxidative damage to cells. The bioinspired architecture not only reduced mechanical and oxidative damage to cells but also maintained normal ROS level for physiological hemostasis. This work provides basic principles to design and develop antioxidative biomaterials for implantation in vivo.

[Mechanism of Mitochondria-mediated Pathway in the Platelet Apoptosis Resulted from Immune Bone Marrow Failure].

OBJECTIVE: To explore the mechamisms of mitochondria-mediated pathway in apoptosis of platelets resulted from in immune induced bone marrow failure. METHODS: Thirty C57BL/6 mice were randomly divided into 3 groups (10 mice in each group): normal group, model group, cyclosporine A(CsA) group. Mouse model of immune bone marrow failure were established. After mouse model was successfully established, the mice in normal group and model group were given saline orally, the mice in CsA group was treated with CsA orally. Blood routine examination of mice in each group was performed by automatic blood cell analyzer; the mitochondrial membrane potential(ΔΨm), cytochrome C(Cyt C), phosphatidylserine (PS), Ca2+ were measured by flow cytometry; expression of BAX, BAK, caspase-3, caspase-8, caspase-9 was detected by using Western blot method, the changes of bone marrow platelet ultrastructure were observed under transmission electron microscope. RESULTS: Compared with normal group, the platelet count of model group decreased significantly, while the level of ΔΨm, caspase-3, caspase-8, caspase-9 significantly decreased, the level of Cyt C, PS, Ca2+, BAX, BAK increased significantly (P<0.05). Compared with the model group, the platelet count of CsA group increased obviously, while the level of ΔΨm, caspase-3, caspase-8, caspase-9 of CsA group increased significantly, the level of Cyt C, PS, Ca2+, BAX, BAK of CsA group decreased significantly (P<0.05). Electron microscopy showed that compared with the model group, platelet damage in CsA group were alleviated. CONCLUSION: mitochondrial pathway plays an important role in the reduction of platelet resulted from immune bone marrow failure.

Flavone from Zhongjiefeng (Herba Sarcandrae Glabrae) inhibits platelet apoptosis in immune-induced bone marrow failure through mitochondrial pathway.

OBJECTIVE: To investigate the effect of Flavone from Zhongjiefeng (Herba Sarcandrae Glabrae) on the platelet number in immune-induced bone marrow failure (BMF) and its mechanism of mitochondrial apoptotic pathway. METHODS: Immune-induced BMF model, established in mice, was randomly divided into four groups: normal control group without BMF, BMF control group, cyclosporine (CSA) group and flavone group (n=10 in each group). Mice were given 0.027 g/kg cyclosporine or 0.2 g/kg flavone lavage daily in either the cyclosporine or flavone group respectively. Platelet count, mitochondrial transmembrane potential, cytochrome C , phosphatidylserine (PS), changes of calcium ion (Ca2+), and protein expression of mitochondrial apoptotic pathway including B-cell lymphoma-2 (bcl-2) Homologous Antagonist-Killer Protein (Bak), bcl-2-associated X protein (Bax), caspase-3, caspase-8, and caspase-9 were examined and compared. RESULTS: Compared with the normal control group, the BMF group had significantly lower levels of platelet count, and expressions of caspase family proteins as well as higher levels of Cyt C, PS, Ca2+, and expressions of Bak and Bax (all P < 0.05). Compared with the BMF group, the CSA and flavone groups had significantly higher and expressions of caspase family proteins (all P<0.05) whereas the levels of Cyt C, PS, Ca2+, and expressions of Bak and Bax were reduced (all P<0.05). More importantly, the flavone group had higher levels of Cyt C, Ca2+ and expressions of Bak and Bax compared with the CSA group (all P<0.05), while the levels of PS and caspase family proteins were reduced (all P<0.05). CONCLUSION: Flavone from Zhongjiefeng (Herba Sarcandrae Glabrae) significantly increases the platelet number and prevents its apoptosis through mitochondrial pathway.

Ultra-high resolution optical coherence tomography for imaging the anterior segment of the eye.

Developments in optical coherence tomography (OCT) have expanded its clinical applications for ultra-high resolution imaging of the anterior segment of the human eye. This review presents the latest advances for imaging the anterior segment of the eye using ultra-high resolution OCT (UHR-OCT). Unique applications of UHR-OCT technology in clinical and basic scientific laboratory research are discussed and a summary of the results is provided. The authors focused on the use of UHR-OCT for imaging of tear dynamics, contact lens interactions with the corneal surface, and in vivo histological diagnosis of disorders of the cornea, as well as the future direction in this field.