Central Retinal Artery Occlusion Due to Subperiosteal Orbital Abscess Caused by Acute Sinusitis in a Child: A Case Report.

Central retinal artery occlusion (CRAO) is an ophthalmic emergency and has poor visual prognosis. It is commonly found in elderly people and very rare in child. We reported an 8-year-old girl who suffered from acute sinusitis, periorbital swelling, and the visual acuity of her right eye was only light perception. She was diagnosed with CRAO, SPOA (subperiosteal orbital abscess), and acute sinusitis. Emergency treatments including surgery, antibiotics, glucocorticoids, intraocular-pressure-lowering drugs, and vasodilators were taken immediately in order to save the eyesight. The visual acuity of the right eye returned to 20/400. Conclusions: Severe intraorbital complications of acute sinusitis can lead to CRAO. Timely drainage, strong antibiotics, and glucocorticoids are the most effective methods for the treatments.

Hypoxia enhances the production and antitumor effect of exosomes derived from natural killer cells.

BACKGROUND: Exosomes are a subgroup of extracellular vesicles that are naturally released by almost all types of cells. However, the factors that promote the capacity of natural killer (NK) cells to release exosomes are unclear. In this study, we investigated whether hypoxia can enhance the yield of NK cell-derived exosomes and improve the immunotherapeutic effects of these cells. METHODS: Exosomes from NK92 or NK92-hIL-15 cells were isolated from culture medium under normoxic (NK92-Exo and NK92-hIL-15-Exo) or hypoxic (hypoxic NK92-Exo and hypoxic NK92-hIL-15-Exo) conditions. NK92-Exo and hypoxic NK92-Exo were characterized by transmission electron microscopy (TEM), nanoparticle-tracking analysis (NTA), and western blot. Real-time cell assay, wound healing assay, flow cytometry, and western blot were then performed to assess cytotoxicity, cell proliferation, cell migration, apoptosis, and the expression levels of cytotoxicity-associated proteins. RESULTS: After 48 hours of hypoxic treatment, NK92-Exo exhibited significantly increased cytotoxicity, enhanced inhibition of cell proliferation, and elevated levels of molecules associated with NK cell cytotoxicity. The hypoxia-treated NK92-Exo and NK92-hIL-15-Exo showed increased expression of three functional proteins of NK cells-specifically FasL, perforin, and granzyme B-as compared with their NK92-Exo counterparts exposed to normoxia. CONCLUSIONS: As an approach that supports overproduction of exosomes, hypoxic treatment of NK cells may serve as a promising therapeutic option for cancer immunotherapy.

Tracking pesticide exposure to operating workers for risk assessment in seed coating with tebuconazole and carbofuran.

BACKGROUND: Coating seed with pesticides is an effective way to control plant pests, however, factory-based coating processes may carry a potential risk to operational workers of chemical exposure. To study the risk, carbofuran and tebuconazole were used to coat corn seed and their subsequent distribution on the bodies of workers was measured at manufacturers XFS and LS (Shanxi, China). Clothing was collected from workers during operations and analyzed using high-performance liquid chromatography. RESULTS: At XFS, dermal exposure to carbofuran was 4.83, 3.31 and 1.48 mg kg-1 , and exposure to tebuconazole was 6.88, 5.16 and 1.72 mg kg-1 for coating, packing and transport workers, respectively. At LS, dermal exposure to carbofuran was 2.32, 0.46 and 0.55 mg kg-1 , and exposure to tebuconazole was 1.69, 0.46 and 0.70 mg kg-1 , for coating, packing and transport workers, respectively. The level of pesticide exposure was significantly higher for seed-coating workers than for packing and transport workers. The main area of exposure was the hands for all workers and the lower limbs for packers; exposure was relatively uniform for pesticide handlers. Occupational risk was assessed based on margin of exposure (MOE). In seed-coating, the MOE was greater than 100 for tebuconazole, indicating no potential risk, but ranged from 0.25 to 2.88 for carbofuran, indicating the risk of a health impact. CONCLUSION: The level of exposure varied depending on type of operation undertaken and body parts of workers' body, but the risk of a health impact was highly associated with pesticide toxicity. This provides a guideline for workers in pesticide manufacturing to ensure safe operation of the seed-coating process. © 2021 Society of Chemical Industry.

hUMSCs regulate the differentiation of ovarian stromal cells via TGF-ß1/Smad3 signaling pathway to inhibit ovarian fibrosis to repair ovarian function in POI rats.

OBJECTIVE: The basic pathological changes of primary ovarian insufficiency (POI) include ovarian tissue fibrosis and follicular development disorders. The human umbilical cord mesenchymal stem cell (hUMSC) transplantation has been shown an effective method to improve the ovarian function in POI rat model; however, the exact mechanisms are still unclear. The purpose of this study is to investigate whether the recovery of ovarian function in POI rats is related to the inhibition of tissue fibrosis following hUMSC transplantation. Furthermore, the transforming growth factor-ß1 (TGF-ß1) signaling pathway is explored to determine the mechanisms of ovarian function recovery through its inhibition of tissue fibrosis. METHODS: The primary ovarian insufficiency (POI) rat model was established by intraperitoneal injection of chemotherapy drug cisplatin (CDDP) for 7 days. The levels of serum sex hormones were measured using enzyme-linked immunosorbent assay (ELISA). The tissue fibrosis in the ovary was examined using Masson staining and Sirius red staining. The collagen fibers in the ovarian tissues were detected by Western blot analysis. To investigate the mechanisms of ovarian function recovery following hUMSC transplantation, ovarian stromal cells were isolated from the ovarian cortex of immature rats. The expression of Cytochrome P450 17A1 (Cyp17a1) and fibrosis marker of alpha smooth muscle actin (α-SMA) in ovarian stromal cells was examined using immunofluorescence analysis. Also, the protein levels of Cyp17a1 and α-SMA in ovarian stromal cells were examined by Western blot analysis. The expression of TGF-ß1 and Smad3 signals was measured by Western blot and quantitative reverse-transcription polymerase chain reaction (qRT-PCR) analysis. RESULTS: The results show that the function of the ovary in POI rats was significantly improved after hUMSC transplantation. The expression of fibrosis markers (α-SMA) and production of Collagen Type I (Collagen I) and Collagen Type III (Collagen III) in POI rats were significantly inhibited in POI rats following hUMSC transplantation. In the cultured ovarian stromal cells, the decrease of TGF-ß1 and p-Smad3 protein expression was observed in hUMSC-treated POI rats. The treatment with TGF-ß1 inhibitor of SB431542 further confirmed this signal pathway was involved in the process. CONCLUSION: Our study demonstrated that the TGF-ß1/Smad3 signaling pathway was involved in the inhibition of ovarian tissue fibrosis, which contributed to the restoration of ovarian function in POI rats following hUMSC transplantation.

Fungicide SYP-14288 Inducing Multidrug Resistance in Rhizoctonia solani.

Rhizoctonia solani is a widely distributed soilborne plant pathogen, and can cause significant economic losses to crop production. In chemical controls, SYP-14288 is highly effective against plant pathogens, including R. solani. To examine the sensitivity to SYP-14288, 112 R. solani isolates were collected from infected rice plants. An established baseline sensitivity showed that values of effective concentration for 50% growth inhibition (EC50) ranged from 0.0003 to 0.0138 µg/ml, with an average of 0.0055 ± 0.0030 µg/ml. The frequency distribution of the EC50 was unimodal and the range of variation factor (the ratio of maximal over minimal EC50) was 46.03, indicating that all wild-type strains were sensitive to SYP-14288. To examine the risk of fungicide resistance, 20 SYP-14288-resistant mutants were generated on agar plates amended with SYP-14288. Eighteen mutants remained resistant after 10 transfers, and their fitness was significantly different from the parental strain. All of the mutants grew more slowly but showed high virulence to rice plants, though lower than the parental strain. A cross-resistance assay demonstrated that there was a positive correlation between SYP-14288 and fungicides having or not having the same mode of action with SYP-14288, including fluazinam, fentin chloride, fludioxonil, difenoconazole, cyazofamid, chlorothalonil, and 2,4-dinitrophen. This result showed a multidrug resistance induced by SYP-14288, which could be a concern in increasing the spectrum of resistance in R. solani to commonly used fungicides.

hUMSC transplantation restores ovarian function in POI rats by inhibiting autophagy of theca-interstitial cells via the AMPK/mTOR signaling pathway.

BACKGROUND: Previous studies of primary ovarian insufficiency (POI) have focused on granulosa cells (GCs) and ignored the role of theca-interstitial cells (TICs). This study aims to explore the mechanism of the protective effects of human umbilical cord-derived mesenchymal stem cells (hUMSCs) on ovarian function in POI rats by regulating autophagy of TICs. METHODS: The POI model was established in rats treated with cisplatin (CDDP). The hUMSCs were transplanted into POI rats by tail vein. Enzyme-linked immunosorbent assay (ELISA) analysis, hematoxylin and eosin (HE) staining, and immunohistochemistry were used to measure the protective effects of hUMSCs. The molecular mechanisms of injury and repairment of TICs were assessed by immunofluorescence, transmission electron microscope (TEM), flow cytometry (FCM), western blot, and quantitative real-time polymerase chain reaction (qRT-PCR). RESULTS: In vivo, hUMSC transplantation restored the ovarian function and alleviated the apoptosis of TICs in POI rats. In vitro, hUMSCs reduced the autophagy levels of TICs by reducing oxidative stress and regulating AMPK/mTOR signaling pathway, thereby alleviating the apoptosis of TICs. CONCLUSION: This study indicates that hUMSCs protected ovarian function in POI by regulating autophagy signaling pathway AMPK/mTOR.

CD83+ CCR7+ NK cells induced by interleukin 18 by dendritic cells promote experimental autoimmune uveitis.

Natural killer (NK) cells have been reported to play a pathological role in autoimmune uveitis. However, the mechanisms regarding NK cells in uveitis and factors that affect NK-cell activation in this condition remain unclear. Here, we report that the number of CD3- NK1.1+ CD83+ CCR7+ cells is increased in the inflamed eyes within a mouse model of experimental autoimmune uveitis (EAU), and these cells express elevated levels of NKG2D, CD69 and IFN-γ. Adoptively transferring CD83+ CCR7+ NK cells aggravates EAU symptoms and increases the number of CD4+ IFN-γ+ T cells and dendritic cells (DCs) within the eye. These CD83+ CCR7+ NK cells then promote the maturation of DCs and IFN-γ expression within T cells as demonstrated in vitro. Furthermore, IL-18, as primarily secreted by DCs in the eyes, is detected to induce CD83+ CCR7+ NK cells. In EAU mice, anti-IL-18R antibody treatment also decreases retinal tissue damage, as well as the number of infiltrating CD83+ CCR7+ NK cells, T cells and DCs in the inflamed eyes and spleens of EAU mice. These results suggest that CD83+ CCR7+ NK cells, as induced by IL-18 that primarily secreted by DCs, play a critical pathological role in EAU. Anti-IL-18R antibody might serve as a potential therapeutic agent for uveitis through its capacity to inhibit CD83+ CCR7+ NK cells infiltration.

NK cells are negatively regulated by sCD83 in experimental autoimmune uveitis.

Natural killer (NK) cells represent a subset of lymphocytes that contribute to innate immunity and have been reported to play a role in autoimmune uveitis. However, the mechanisms regulating NK cellular function in this condition remain unclear. Herein, we investigated the status of NK cells in experimental autoimmune uveitis (EAU). We found that the number of CD83+CD3-NK1.1+ cells was increased in the inflamed eyes and spleens of the EAU mouse model. At the recovery stage of EAU, serum concentrations of soluble CD83 (sCD83) were increased. sCD83 treatment relieved retinal tissue damage and decreased the number of infiltrating NK cells in inflamed eyes. Further analysis of the effects of sCD83 treatment in EAU revealed that it reduced: 1) the expressions of CD11b and CD83 in NK cells, 2) the percent of CD11bhighCD27lowCD3-NK1.1+ cells and 3) the secretion of granzyme B, perforin and IFN-γ in NK cells as demonstrated both in vivo and in vitro. When sCD83 treated-NK cells were transferred into EAU mice, retinal tissue damage was relieved. These results demonstrate sCD83 down-regulate NK cellular function and thus provide important, new information regarding the means for the beneficial effects of this agent in the treatment of autoimmune uveitis.

Human decidua mesenchymal stem cells regulate decidual natural killer cell function via interactions between collagen and leukocyte­associated immunoglobulin­like receptor 1.

The development of maternal tolerance to the fetal allograft in critical for the maintenance of the pregnancy, and it is accompanied by the development of a special decidual natural killer (dNK) cell tolerance phenotype. To understand the factors that influence dNK cells during early pregnancy, the present study aimed to identify mesenchymal stem cells (MSCs) from human first­trimester deciduas, termed decidual MSCs (DMSCs), and to investigate the effect of DMSCs on the regulation of dNK cells via collagen. Decidual samples were collected from women with normal pregnancy that had undergone elective vaginal surgical terminations at 6­9 weeks gestation. DMSCs derived from human decidual tissues were cultured under differentiation conditions to examine their multipotent differentiation capacities, and the expression of MSC­specific markers, including cluster of differentiation (CD)44, CD73, CD105, CD90, CD34, CD31, CD14, CD45, CD11b and human leukocyte antigen­antigen D related, was determined. dNK cells were co­cultured with DMSCs in order to examine the effect of DMSCs on the tolerance phenotype of dNK cells. The expression of cell surface molecules, natural cytotoxicity triggering receptor 3 and killer cell immunoglobulin­like receptor (KIR) 2DL1, and the secretion of cytokines, including interferon­Î³, tumor necrosis factor (TNF)­α, interleukin (IL)­10, IL­4 and perforin, were examined by flow cytometry analysis. To determine whether the regulation of dNK cells by DMSCs was mediated by collagen, DMSCs were pre­treated with human recombinant leukocyte­associated immunoglobulin­like receptor (LAIR)­2 and transfected with pScoR­GFP­hP4H to inhibit the interaction between LAIR­1 and collagen. The present results demonstrated that collagen produced by DMSCs increased the expression of KIR2DL1 and IL­4, decrease the expression of NKp30 and TNF­α. In conclusion, the results of the present study demonstrated that DMSCs may be cultured in vitro for prolonged periods, whilst retaining the ability to differentiate into different cell lineages. In addition, DMSCs may modulate the function of dNK cells via the interaction between collagen and LAIR­1.

Increased serum mannose binding lectin levels are associated with diabetic retinopathy.

BACKGROUND: Mannose-binding lectin (MBL) may be implicated in the pathogenesis of diabetic retinopathy (DR) complications. We investigated serum MBL levels in type 2 diabetic patients with and without DR. METHOD: Serum MBL levels were determined in 184 type 2 diabetic patients with DR and 189 type 2 diabetic patients without DR matched for age, sex, and duration of diabetes. Multivariate analyses were performed using logistic regression models. The diagnostic value of MBL was compared with the HbA1c, Hs-CRP and with other known markers. RESULTS: We found that serum MBL levels were significantly higher in diabetes with DR as compared to without-DR [3456 (IQR, 3128-3800) ug/l and 2432 (IQR, 2100-2670) ug/l, respectively; P<0.0001]. In multivariate logistic regression analysis, after adjusting for all other significant factors, MBL remained can be seen as an independent DR marker with an adjusted OR of 1.002 (95% CI, 1.001-1.003; P<0.0001). Based on the ROC curve, the optimal cutoff value of serum MBL levels as an indicator for diagnosis of DR was projected to be 3050 ug/L, which yielded a sensitivity of 82.5% and a specificity of 88.0%, with the area under the curve at 0.907 (95% CI, 0.876-0.938). CONCLUSION: In type 2 diabetic patients, evaluated serum levels of MBL can be seen as an independent marker of DR even after correcting for possible confounding factors.

Synergistic protective effects of escin and low­dose glucocorticoids against vascular endothelial growth factor­induced blood­retinal barrier breakdown in retinal pigment epithelial and umbilical vein endothelial cells.

Previous studies have shown that escin possesses glucocorticoid (GC)­like anti­edematous and anti­inflammatory effects. The present study was designed to investigate whether escin exhibits synergistic protective effects against blood­retinal barrier (BRB) breakdown when combined with GC in an in vitro monolayer BRB model, based on retinal pigment epithelial (RPE) cells and human umbilical vein endothelial cells (HUVECs). The results showed that low concentrations of escin and triamcinolone acetonide (TA) administered separately did not affect BRB trans­endothelial (epithelium) resistance (TEER). However, when administered together, escin and TA significantly inhibited reduced BRB TEER following treatment with vascular endothelial growth factor (VEGF). Furthermore, low­concentrations of escin and TA administered together significantly increased the expression levels of occludin and ZO­1. This demonstrates that escin and GC have synergistic protective effects against BRB breakdown, and the molecular mechanisms may be related to the upregulation of occludin and ZO­1 expression. The combination of escin with GC indicates a potential beneficial strategy for the treatment of breakdown of the BRB.

Effects of hydroxysafflor yellow A on proliferation and collagen synthesis of rat vascular adventitial fibroblasts induced by angiotensin II.

THE PURPOSE OF THIS STUDY WAS TO: 1) examine the effects of hydroxysafflor yellow A (HSYA) on the proliferation, collagen and cytokine synthesis of vascular adventitial fibroblasts as induced by angiotensin II (Ang II) in normal Sprague-Dawley (SD) rats in vitro, and 2) to assess the effects of HSYA on morphological changes and collagen accumulation of vascular adventitia in spontaneously hypertensive rats (SHR) in vivo. In vitro experiment, vascular adventitial fibroblasts from SD rats were isolated, cultured, and divided into control groups, model groups and HSYA groups. Cell morphology of adventitial fibroblasts was assessed using laser confocal microscopy, while cell proliferation with the MTT assay, and collagen synthesis was determined using hydroxyproline chromatometry. Immunocytochemistry and reverse transcription PCR were used for detecting the expression of TGF-ß1, MMP-1, α-SMA and NF-κB in adventitial fibroblasts. In vivo experiment, vascular adventitia proliferation and collagen synthesis were analyzed using hematoxylin-eosin and Sirius staining. Our results showed that: 1) in vitro experiment of SD rats, HSYA inhibited proliferative activity and collagen synthesis of adventitial fibroblasts as induced by Ang II, and the inhibitory effects of HSYA on the increased expression of MMP-1, TGF-ß1, α-SMA and NF-κB p65 as induced by Ang II were assessed, and 2) in vivo experiment of SHR, histological analysis displayed fewer pathological changes of vascular adventitia in HSYA treatment groups as compared with no HSYA treatment groups, and MMP-1, TGF-ß1, α-SMA and NF-κB p65 expression significantly reduced after HSYA treatment (P < 0.05). Our results revealed that HSYA treatment significantly decreased the amount of cytokines and collagen synthesis in vascular adventitia components. This study provides experimental evidence demonstrating that HSYA has the capacity to decrease vascular adventitia proliferation and hyperplasia during vascular remodeling.

Effects of BMSCs interactions with adventitial fibroblasts in transdifferentiation and ultrastructure processes.

In this study an in vitro model of simulated blood vessel injury was used to study the effects of bone marrow-derived mesenchymal stem cells (BMSCs) morphology and to detect vascular smooth muscle actin (SM α-actin) expression in the presence of adventitial fibroblasts. BMSCs from rats with DAPI-labeled nuclei were co-cultured with adventitial fibroblasts for 7 days, while BMSCs cultured alone served as controls. Cell morphology of BMSCs was assessed by laser confocal microscopy and SM α-actin or calponin expression in BMSCs was detected by immunofluorescence staining. The expression of SM α-actin mRNA was identified using RT-PCR. Cell ultrastructure was assessed by electron microscopy. The results demonstrate that BMSCs with DAPI-labeled nuclei were smaller compared with fibroblasts, and their nuclei emitted a blue fluorescence. Most BMSCs displayed a polygonal shape changing from their original long fusiform shape. BMSCs with blue nuclei and red cytoplasm (SM α-actin positive or calponin positive) were observed, and a substantial number of filaments were present in the cytoplasm as observed under electron microscopy. The number of these cells increased as a function of culture duration. However, SM α-actin expression was weak and calponin expression was not detected in the control group. This study provides important new information on the characterization of artherosclerosis pathogenesis and vascular restenosis after blood vessel injury. Our findings demonstrate that direct interactions with adventitial fibroblasts can induce vascular smooth muscle-like cell differentiation in BMSCs.