Ocular microvascular alteration in Sjögren syndrome.

BACKGROUND: Sjögren syndrome (SjS) is a systemic disease affecting exocrine, including ocular lacrimal, glands. It is uncertain whether ocular microvascular alterations are associated with this disease. In this study, we evaluated retinal and conjunctival microvascular changes in SjS patients using optical coherence tomography angiography (OCTA). METHODS: Twelve SjS patients (24 eyes) and 12 normal controls (24 eyes) were recruited to this study. Three-dimensional conjunctival and retinal OCTA images of each eye were captured and microvascular density was calculated. Each image was analyzed by retinal area based on the early treatment of diabetic retinopathy study method (R, S, L, and I) hemisphere segmentation method (SR, SL, IL, and IR); and central wheel division method (C1-C6). Correlation analyses were used to look for associations between retinal and conjunctival microvascular densities. RESULTS: Superficial and deep retinal layer microvascular density was decreased in SjS patients compared with normal controls (P<0.05). This significant difference was found in both superficial and deep layers in S, L, SL, IL and C1-C3 regions, and additionally in the I and SR regions in the superficial layer. Conversely, in the conjunctiva microvascular density was higher in SjS patients than in controls. In SjS patients, a significant negative correlation was found between conjunctival and both superficial (r=-0.641; P=0.025) and deep (r=-0.958; P<0.0001) microvascular densities. CONCLUSIONS: The changed microvascular densities measured in deep and superficial retinal layers and in the conjunctiva demonstrate that OCTA is a promising method in differentiating the eyes from those with SjS.

[Effects of tillage managements on soil microbial community structure in soil aggregates of fluvo-aquic soil]. / 耕作方式对潮土土壤团聚体微生物群落结构的影响.

In order to explore the impacts of different tillage managements on the structure and diversity of microbial community in fluvo-aquic soil, the phospholipid fatty acid (PLFA) method was used to determine microbial community composition in soil aggregates. Four tillage treatments were set up in Qihe County, Shandong Province, including rotary tillage with straw return (RT), deep ploughing with straw return (DP), subsoiling with straw return (SS) and no-tillage with straw return (NT). Our results showed that DP treatment significantly increased the amount of fungal PLFAs and fungi/bacteria ratio in >5 mm soil aggregates compared with RT. DP could provide favorable conditions for fungi reproduction, facilitate soil organic matter storage and soil buffering capacity. DP increased the amount of PLFAs in 5-2 mm soil aggregates, reduced the gram-positive (G+) /gram-negative (G-) bacteria ratio in the soil, and improved soil nutritional status. In addition, DP improved the microbial abundance index in <0.25 mm soil aggregates. In general, DP could not only increase the abundance of bacteria and fungi in soil aggregates, but also improve the microbial community structure of soil aggregate, which help increase soil carbon sequestration capacity and keep soil microbial diversity to a certain extent. Results of the redundancy analysis showed that the total PLFAs, PLFAs of bacteria, G- bacteria and actinomycetes in soil aggregates are closely correlated with soil organic carbon, while PLFAs of G+ bacteria had a strong correlation with soil total nitrogen concentration. In each treatment, microbial communities in larger sizes of soil aggregates were mainly affected by the ratio of organic carbon to total nitrogen, soil moisture, pH, and mass fractions of soil aggregates, while the microbial communities in smaller sizes of soil aggregates were affected by the concentrations of organic carbon and total nitrogen.

Brain Functional Connectivity Changes in Patients with Acute Eye Pain: A Resting-State Functional Magnetic Resonance Imaging (fMRI) Study.

BACKGROUND By using functional magnetic resonance imaging (fMRI), we aimed to study the changes in potential brain function network activity in patients with acute eye pain. Also, by using the voxel-wise degree centrality (DC) method, we aimed to explore the relationship between spontaneous brain activity and the clinical features of patients with acute eye pain. MATERIAL AND METHODS A total of 15 patients with acute eye pain (5 women and 10 men; EP group) and 15 healthy controls (5 women and 10 men; HC group), were scanned by fMRI. The DC method was used to evaluate changes in spontaneous brain activity. Receiver operating characteristic (ROC) curves were analyzed, and Pearson correlation analysis was used to study the relationship between DC values and clinical manifestations in different regions of brain. RESULTS The area of the left limbic lobe showed a reduction in DC value in patients in the EP group. DC values were elevated in the left cerebellum posterior lobe, left inferior parietal lobule, left inferior temporal gyrus, left precuneus, and right cerebellum posterior lobe in the EP group. The visual analog scale value of the eyes in the EP group was negatively correlated with the left limbic lobe signal value and positively correlated with the left inferior parietal lobule signal value. Further, the scores of the hospital anxiety and depression scale and DC value of the left limbic lobe were negatively correlated. CONCLUSIONS Compared with the HC group, patients with acute eye pain had abnormal patterns of intrinsic brain activity in different brain regions, which may help reveal the potential neural mechanisms involved in eye pain.

Altered Spontaneous Brain Activity Patterns in Patients After Lasik Surgery Using Amplitude of Low-Frequency Fluctuation: A Resting-State Functional MRI Study.

OBJECTIVE: Previous studies demonstrated that myopia could result in alterations of brain activity in specific areas. However, whether the visual function could improve by Lasik surgery, with the brain activity alterations also change, is still unknown. Here, we intended to use the amplitude of low-frequency fluctuation (ALFF) technique to investigate the intrinsic brain activity changes in pre-Lasik (PRL) and post-Lasik (POL) patients. METHODS: A total of 15 patients with myopia (nine male and six female) were recruited in our study, who were matched according to age, weight, and height. These patients comprised both the PRL and POL groups, which is self-controlled. The patients all underwent resting-state functional magnetic resonance imaging (MRI), and the spontaneous brain activity changes were recorded by the ALFF technique. The data were recorded and arranged in the receiver operating characteristic (ROC) curve, which presented how intrinsic activities altered in different brain regions. Moreover, by Graphpad prism 8, we can analyze the linear correlation between HADS (Hospital Anxiety and Depression Scale) and ALFF values as well. RESULTS: Differences in ALFF values existed in brain regions between the same patient before and after the Lasik operation. The regions with increased ALFF values after Lasik surgery were the left parahippocampal gyrus, cerebellar vermis, and left posterior cingulate cortex. The regions with decreased ALFF values after Lasik surgery were the left supramarginal gyrus and right trigonometric inferior frontal gyrus. CONCLUSION: We demonstrated significant fluctuations of ALFF values in specific brain areas between the same patients before and after the Lasik surgery. The altered ALFF values reflected the hyperactivity or hypoactivity of the specific brain areas, which may help predict the recovery level of patients' vision after Lasik surgery. Furthermore, based on the experimental results that presented significant activity alterations in specific brian regions, patients could be speculated equipped with a better visual function.

Netrin-1 promotes epithelium repair in corneal injury.

AIM: To explore netrin-1 functions on corneal epithelium in vitro and in vivo. METHODS: In vitro the human corneal epithelial (HCE) cells were treated with serum free DMEM-F12 basic media containing 0, 50, 100, 200, 300, 500, 800, and 1000 ng/mL of netrin-1, respectively. The cells viability was detected by cell counting kit-8 (CCK-8). The wound-healing assay was applied to assess the migration proficiency of HCE cells. Flow cytometry was used to analyze the cell-cycle distribution and apoptosis. In vivo, normal c57 (6wk) mice were demarcated with a trephine in the middle of the cornea to produce a 3-mm circular wound. Mice corneas were inflicted no epithelium with a 3-mm wound displayed, but remained the limbal epithelium intact. A blunt scalpel blade was used to remove the corneal epithelian cells, followed by topical netrin-1 application (200 ng/mL), and the group treated by PBS as control. The treated group was injected netrin-1 into the normal c57 mice inferior subconjunctival 4h before trauma. Mouse corneal inflammation and neovascularization were observed under slit lamp microscope. The apoptosis of corneal cells was determined by TUNEL staining. RESLUTS: A concentration of 200 ng/mL netrin-1 enhanced 25% of the HCE viability. The relative migration rates were 76.3% and 100% in control and netrin-1 treated group after cultured 72h. Treated with netrin-1 (200 ng/mL) decreased the apoptosis of HCE cells, as well as decreased their percentage from 19.3%±0.57% to 12.7%±0.42% of the total. The remaining wound area was 1.22 mm2 in control group but 0.22 mm2 in the netrin-1 treated group. Exogenous Netrin-1 inhibits apoptosis of corneal epithelial cells of c57 mice. TUNEL-positive cells at the epithelial layer of the corneas of the control and netrin-1 treated c57 mice at 24h after wounding were 43.3% and 16.7% respectively. CONCLUSION: Netrin-1 can reduce HCE apoptosis as well as promote its proliferation and migration. Topical application of netrin-1 promotes the injuryed cornea epithelial wound repair and inhibits apoptosis of corneal epithelial cells. These findings may offer potential therapies to repair the defects of corneal epithelial based on netrin-1.

Altered brain network centrality in patients with late monocular blindness: a resting-state fMRI study.

INTRODUCTION: The aim of the study was to investigate the underlying functional network brain activity changes in patients with late monocular blindness (MB) and the relationship with their clinical features using the voxel-wise degree centrality (DC) method. MATERIAL AND METHODS: A total of 32 patients with MB (25 males and 7 females), and 32 healthy controls (HCs) (25 males and 7 females) closely matched in age, sex, and education, underwent resting-state functional magnetic resonance imaging scans. The DC method was used to assess local features of spontaneous brain activity. Correlation analysis was performed to explore the relationships between the observed mean DC signal values of the different areas and clinical features in these patients. RESULTS: Compared with HCs, MB patients had significantly lower DC values in the bilateral cuneus/V1/V2, and significantly higher DC values in the left inferior temporal gyrus and bilateral medial frontal gyrus. However, there was no relationship between the observed mean DC values of the different brain areas and the behavioral performance. CONCLUSIONS: Late monocular blindness involves brain function network dysfunction in many regions, which might indicate impairment of the visual cortex and other vision-related brain regions in the MBs.

Reduction in interhemispheric functional connectivity in the dorsal visual pathway in unilateral acute open globe injury patients: a resting-state fMRI study.

This study investigated the changes in interhemispheric functional connectivity (FC) of the whole brain in open globe injury (OGI) patients, using voxel-mirrored homotopic connectivity (VMHC), and their relationships with clinical features. Totally, 16 male and 2 female acute OGI patients and 18 sex, age, and education-matched healthy volunteers were enrolled in the study. All subjects were scanned through functional magnetic resonance imaging (fMRI). Receiver operating characteristic (ROC) curves analyses had been used to identify the VMHC in these brain areas could be used as biomarkers to distinguish OGI and from healthy control (HC). The mean VMHC values in multiple brain areas and clinical OGI manifestations were evaluated with a Pearson correlation analysis. OGI patients had significantly decreased VMHC in the bilateral calcarine/lingual/cuneus (BA18, 19, 30) and middle occipital gyrus (BA18, 19). The OGI patients had abnormal interhemispheric FC in the dorsal visual pathway, which may represent the pathophysiological mechanism that underlies acute vision loss after OGI.

Resting cerebral blood flow alterations specific to the comitant exophoria patients revealed by arterial spin labeling perfusion magnetic resonance imaging.

PURPOSE: It has been shown in many previous studies that there were significant changes of the brain anatomy and function in strabismus. However, the significance of the alterations of resting cerebral blood flow (CBF) in comitant exophoria (CE) remains obscure. Arterial spin labeling (ASL) MRI, which is a noninvasive method, could be applied to detect the cerebral blood flow quantitatively. Our study aimed to compare the resting CBF between the comitant exophoria and health controls using pseudo-continuous arterial spin labeling (pCASL) perfusion MRI method. METHODS: 32 patients (25 males and 7 females) with CE (study group), and 32 (25 males and 7 females) healthy individuals with matched age and sex status (control group) underwent a whole-brain pCASL magnetic resonance (MR) examination at the resting state. The resting CBF were voxel-wise compared between the two groups using an analysis of variance designed in a statistical parametric mapping program. The CE patients were distinguishable from the healthy controls (HCs) by receiver operating characteristic (ROC) curves. RESULTS: Compared with the control group, the CE group showed significantly increased resting CBF values in the right parahippocampal regions, bilateral medial frontal gyrus/anterior cingulate cortex, left inferior frontal gyrus, right inferior frontal gyrus, left superior frontal gyrus, bilateral medial cingulate cortex, right middle frontal gyrus, and right paracentral lobule. CONCLUSION: Comitant exophoria showed increased resting CBF in eye movement-related brain areas including supplementary eye field, cingulate eye field and frontal eye field, which could be an explanation of the brain function compensation for the ocular motility disorders in the CE patients.

Xanthatin inhibits corneal neovascularization by inhibiting the VEGFR2­mediated STAT3/PI3K/Akt signaling pathway.

Alkali burn is one of the main causes of corneal injury. The inflammation and neovascularization caused by alkali burns aggravate corneal damage, resulting in loss of vision. The aim of the present study was to evaluate the efficacy of xanthatin in the treatment of alkali burn­induced inflammation and neovascularization. A CCK­8 assay was used to detect the effects of different concentrations of xanthatin on the proliferation of human umbilical vein endothelial cells (HUVECs). The effects of xanthatin on the migration of HUVECs and the ability of lumen formation were examined using a scratch test and lumen formation assay, respectively. A total of 60 Sprague­Dawley rats were randomly divided into two groups to establish a corneal alkali burn model, and were treated with PBS and xanthatin eye drops four times a day. A slit lamp microscope recorded changes of the cornea at 0, 4, 7, 10 and 14 days, and the inflammatory indices of the cornea and the neovascular area were evaluated. The expression levels of vascular endothelial growth factor (VEGF) and pigment epithelium­derived factor (PEDF) in the cornea under different treatment conditions were detected using immunofluorescence and western blot analysis. In order to investigate the mechanism of xanthatin on the inhibition of inflammation and neovascularization, HUVECs were treated with xanthatin and PBS following VEGF treatment. The subcellular localization of signal transducer and activator of transcription 3 (STAT3) was detected using immunofluorescence. The expression levels of VEGF receptor 2 (VEGFR2), STAT3, phosphoinositide 3­kinase (PI3K) and Akt were detected using western blot analysis. The results revealed that xanthatin inhibited the proliferation of HUVECs in a concentration­dependent manner. The migration ability and lumen­forming ability of the HUVECs were also inhibited by xanthatin. Slit lamp microscopy showed that the inflammatory index and the area of neovascularization in the xanthatin­treated group were significantly reduced, compared with those in the PBS treatment group. The xanthatin treatment group exhibited a lower protein expression level of VEGF and increased protein expression level of PEDF, compared with the PBS treatment group. In the VEGF­treated HUVECs, xanthatin significantly decreased the expression levels of p­VEGFR2, phosphorylated (p­)STAT3, p­PI3K and p­Akt. In conclusion, the present study confirmed that xanthatin inhibited corneal neovascularization and inflammation in the alkali burn model, elucidating the underlying mechanisms involved in its protective effects. Therefore, xanthatin may be a novel drug for the treatment of corneal alkali burn.

A murine model of dry eye induced by topical administration of erlotinib eye drops.

In the present study, the effects of erlotinib on mouse tear function and corneal epithelial tissue structure were investigated. Throughout the 3 weeks of treatment, no notable differences were observed in the body, eye or lacrimal gland weights of the control and experimental mice. However, in the experimental group, the tear volume and break­up times of tear film were significantly lower following treatment with erlotinib compared with the control group. Corneal fluorescein staining in the experimental group revealed patchy staining, and the Lissamine green staining and inflammatory index were significantly higher in the experimental group at 3 weeks than in the control group. In the experimental group, the number of corneal epithelium layers increased significantly following treatment with erlotinib for 3 weeks and a significant increase in the number of vacuoles was observed compared with the control group. Treatment with erlotinib significantly increased the corneal epithelial cell apoptosis, and led to a significantly increased number of epithelial cell layers and increased keratin 10 expression. It also significantly reduced the number of conjunctival goblet cells. Transmission electron microscopy and scanning electron microscopy revealed that the corneal epithelial surface was irregular and there was a substantial reduction and partial loss of the microvilli in the experimental group. Mice treated with erlotinib also exhibited an increased protein expression of tumor necrosis factor­α and decreased protein expression of phosphorylated­epidermal growth factor receptor in the corneal epithelial cells. The topical application of erlotinib eye drops was revealed to induce dry eyes in mice. This is a novel method of developing a model of dry eyes in mice.

Antiangiogenic effects of catalpol on rat corneal neovascularization.

To investigate the effects of catalpol on corneal neovascularization (CNV) and associated inflammation, eye drops (5 mM catalpol or PBS) were administered four times daily to alkali­burn rat models of CNV and inflammation. Clinical evaluations of CNV and the degree of inflammation were performed on days 0, 4, 7, 10 and 14 under slit lamp microscopy. Eyes were collected on day 14 and prepared for hematoxylin and eosin, and immunofluorescence staining; corneal cell apoptosis was investigated via terminal deoxynucleotidyl transferase­mediated nick end labeling (TUNEL) staining. Protein expression levels of angiogenic and proinflammatory factors, including vascular endothelial growth factor (VEGF), pigment epithelium­derived factor (PEDF), tumor necrosis factor­α (TNF­α) and necrosis factor­κB (NF­κB) were determined by western blotting. The effects of catalpol on cell proliferation were investigated in vitro using human umbilical vein endothelial cells (HUVECs) and a Cell Counting kit­8 (CCK­8); alterations in migration and tube formation were investigated via HUVEC wound closure and tube formation assays. HUVEC viability and proliferative ability were inhibited in a dose­dependent manner; catalpol also decreased HUVEC cell migration and tube forming ability. Within alkali­burn rat models, decreased inflammation and CNV was associated with catalpol administration; as demonstrated with TUNEL, corneal cell apoptosis was decreased in response to catalpol. Western blot analysis revealed reduced protein expression levels of VEGF and TNF­α; however, PEDF and phosphorylated­NF­κB p65 were increased due to catalpol administration. The present study demonstrated the inhibitory effects exerted by catalpol on CNV and inflammation within alkali­burned rat models. Topical application of catalpol in vivo was associated with reduced CNV and inflammation; therefore, catalpol may be considered an anti­inflammatory agent for the clinical treatment of CNV.

Retinal microvasculature alteration in central serous chorioretinopathy.

The aim of the present study was to investigate the changes of retinal microvascular network in patients with central serous chorioretinopathy (CSCR). A total of fifteen patients (right eye) with CSCR and 15 normal controls (right eye) were recruited. We used optical coherence tomography angiography to scan 6x6 mm macular retinal blood flow images with the application of a series of customized image segmentation processing program software to obtain microvascular and macrovascular density, and compared the superficial microvascular (SMIR), superficial macrovascular ring (SMAR) and the superficial total microvascular (STMI) density between CSCR patients and control group. Using the annular partition (C1­C6) and quadrant partition methods on the macular, we compared the retinal vessel density changes. We also performed ROC analysis of superficial retinal microvessel density in CSCR retina to investigate the relationship between the microvascular density, the foveal thickness and visual acuity. The density of STMI and SMIR decreased in macular area in the patients with CSCR compared to the normal controls (P<0.05), while the density of SMAR did not change significantly. We found no significant difference in the density of SMIR with the quadrant partition method, whereas the annular partition method showed significantly decreased SMIR density only in the C1 region in patients with CSCR (P<0.05), with no significant difference observed in C2­C6 regions. The density of SMIR had the highest differentiation power in the CSCR group, whereas the density of SC1 ring had the lowest differentiation power by the annular method. The largest area under the ROC curves was 0.77. The correlation index of the SMIR density and visual acuity was ­0.544, whereas macular thickness and visual acuity was ­0.644 in the CSCR group. The density of STMI and SMIR were decreased in patients with CSCR, which might provide further understanding of the pathogenesis of CSCR.

Effect of mistletoe combined with carboxymethyl cellulose on dry eye in postmenopausal women.

AIM: To investigate the protective effect of mistletoe combined with carboxymethyl cellulose eye drops on dry eye in postmenopausal women. METHODS: Sixty postmenopause female patients diagnosed of dry eye were assigned randomly to mistletoe combined with carboxymethyl cellulose eye drops treatment group (n=30) and control group treated with normal saline eye drops (n=30). The subjective symptoms of ocular surface, Ocular Surface Disease Index (OSDI), tear film function tests, tear protein and corneal morphology by confocal scanning microscopy were analyzed before treatment and at 1, 2, 4 and 8wk after treatment respectively. To ensure the safety of the trial, all patients were examined with systolic pressure, diastolic pressure, glutamic-pyruvic transaminase, glutamic oxaloacetic transaminase, urine creatinine, and blood urea nitrogen at 8wk after treatment. RESULTS: There were no obvious differences between two groups before the treatment (P>0.05). In two months after the treatment, the symptoms of ocular surface, OSDI, tear protein, and tear film function were only slightly changed in normal saline eye drops group. However, all indices were improved after the treatment of mistletoe combined with carboxymethyl cellulose eye drops group (P<0.05). In addition, the average amount of corneal epithelium basal cells and inflammatory cells of mistletoe treated group were 3174±379 and 38±25 cells/mm2, significantly decreased as compared to the control group with 4309±612 and 158± 61 cells/mm2, respectively. In the control group, although nerves still maintained straight under corneal epithelium, the number of nerves were significantly decreased, as compared with normal female. In the mistletoe treated group, the number of nerves was only slightly reduced, compared with normal female. CONCLUSION: Mistletoe combined with carboxymethyl cellulose eye drops can alleviate the symptoms and signs of dry eye symptoms.

Network centrality in patients with acute unilateral open globe injury: A voxel­wise degree centrality study.

The present study aimed to investigate functional networks underlying brain­activity alterations in patients with acute unilateral open globe injury (OGI) and associations with their clinical features using the voxel­wise degree centrality (DC) method. In total, 18 patients with acute OGI (16 males and 2 females), and 18 healthy subjects (16 males and 2 females), closely matched in age, sex and education, participated in the present study. Each subject underwent a resting­state functional magnetic resonance imaging scan. The DC method was used to assess local features of spontaneous brain activity. Receiver operating characteristic curve analysis was used to distinguish OGIs from healthy controls (HCs). Correlation analysis was used to examine the association between the observed mean DC values of different brain areas and behavioral performance. Compared with HCs, patients with acute unilateral OGI had significantly increased DC values in the bilateral primary visual cortex (V1/V2) and left precuneus (PCUN), and significantly decreased DC values in the right insula, left insula, right inferior parietal lobule (IPL)/supramarginal gyrus (SMG), IPL/SMG, right supplementary motor area and right postcentral gyrus. Additionally, in the acute OGI group, it was observed that the duration of OGI was negatively correlated with the DC signal value of the bilateral V1/V2 (r=­0.581; P=0.011) and left PCUN (r=­0.508; P=0.031). Acute OGI led to brain functional network dysfunction in a number of brain regions, which may indicate impairment of the visual cortex and other vision­associated brain regions in OGI.

Altered regional homogeneity in patients with late monocular blindness: a resting-state functional MRI study.

Many previous studies have demonstrated that the blindness patients have has functional and anatomical abnormalities in the visual and other vision-related cortex. However, changes in the brain function in late monocular blindness (MB) at rest are largely unknown. In this study, we investigated the underlying regional homogeneity (ReHo) of brain-activity abnormalities in patients with late MB and their relationship with clinical features. A total of 32 patients with MB (25 male and seven female) and 32 healthy controls (HCs) (25 male and seven female) closely matched in age, sex, and education underwent resting-state functional MRI scans. The ReHo method was used to assess local features of spontaneous brain activities. Patients with MB were distinguishable from HCs using the receiver operating characteristic curve. The relationship between the mean ReHo in brain regions and the behavioral performance was calculated using correlation analysis. Compared with HCs, patients with MB showed significantly decreased ReHo values in the right rectal gyrus, right cuneus, right anterior cingulate, and right lateral occipital cortex and increased ReHo values in the right inferior temporal gyrus, right frontal middle orbital, left posterior cingulate/precuneus, and left middle frontal gyrus. However, there was no significant relationship between the different mean ReHo values in the brain regions and the clinical features. Late MB involves abnormalities of the visual cortex and other vision-related brain regions, which may reflect brain dysfunction in these regions.