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Differentiated cells can be reprogrammed to embryonic stem cell-like cells called induced pluripotent stem cells (iPSCs), in which the natural developmental differentiation process is reversed. It is unclear whether the multi-lineage cells can be isolated and identified during reprogramming. In the current study, we detected the expression of lineage markers, isolated neural lineages, and identified the related microRNAs during iPSC formation. Our results demonstrated that a neuroectoderm appeared earlier than mesoderm and definitive endoderm before forming colonies when mouse embryonic fibroblasts were subjected to iPSC formation using transcription factors (TFs). On day 3, the cells expressed Sox1 and Nestin and had ultrastructure consistent with the transition to identity neural germ layer lineage. Fluorescence-activated cell sorting analysis revealed a peak (40%) in neural progenitor marker-positive cells. When subsequently cultured in a neural precursor cell medium, these cells proliferated slowly, became round and aggregated, generating into neurons and glia. Genome-wide microRNA (miRNA) analysis identified 45 differentially regulated miRNAs. Molecular network analysis demonstrated that these miRNAs validated 6,047 experimental mRNA targets. The GO functional annotation analysis of mRNA targets revealed that most genes were related to neurogenesis, such as growth cone, neuronal cell body, neuron projection, and cell junction synapse. The network of protein-protein interactions was observed, which demonstrated that key nodes of neural lineage reprogramming-associated targets were Sall1, Foxa2, Nf2, Ctnnb1, Shh, and Bmpr1a. Therefore, these data suggested that TFs can drive the reprogramming of somatic cells towards a pluripotent state via neuroectoderm. Moreover, the neural lineage reprogramming system can address how miRNAs influence their target sites.

The long non-coding RNA GAS5 (GAS5) is reportedly implicated in glaucoma. However, its significance in human trabecular meshwork cells (HTMCs) remains largely unclear. Here, we investigated the effect of GAS5 on the function of HTMCs and its interaction with miR-29b-3p in HTMCs. We established an H2O2-induced oxidative injury model using HTMCs. RT-qPCR or western blotting was performed to examine the expression of the indicated genes. Luciferase reporter assay was used to determine the interaction between GAS5, miR-29b-3p, miR-29b-3p, and STAT3. CCK8 assay was used to assess the proliferative rate of HTMCs. Exposure to H2O2 increased the expression of Bax, cleaved caspase-3, and extracellular matrix (ECM) proteins, accompanied by reduced Bcl-2 expression. These H2O2-induced changes were effectively alleviated by GAS5 knockdown with sh-GAS5. MiR-29b-3p was directly regulated by GAS5. The effect of sh-GAS5 on ECM protein expression was also observed with the miR-29b-3p mimic. STAT3 was directly regulated by miR-29b-3p. MiR-29b-3p silencing alleviated STAT3 inhibition, followed by the restoration of cell vitality, Bax, Bcl-2, and cleaved caspase-3 expression, and ECM deposition. Our study is the first experimental investigation to shed light on a novel molecular mechanism of the GAS5/miR-29b-3p/STAT3 axis in an H2O2-induced oxidative injury model using HTMCs, which may offer a promising therapeutic approach against glaucoma.

Porcine circovirus 2 (PCV2) causes immunosuppression. Piglets infected with PCV2 can develop enteritis. Given that the gut is the largest immune organ, however, the response of the gut's immune system to PCV2 is still unclear. Here, IPEC-J2 cells with different treatments were co-cultured with PBMC or CD4

PurposeComitant exotropia (CE) is a common eye disorder characterized by impaired stereoscopic vision and eye deviation. Previous neuroimaging studies demonstrated that patients with CE were accompanied by specific functional and structural abnormalities of the brain. However, the effect of impaired stereoscopic vision and eye deviation on interhemispheric homotopic connectivity remains unknown.MethodsA total of thirty-six patients with CE (25 males and 11 females) and 36 well-matched healthy controls underwent magnetic resonance imaging scanning. The voxel-mirrored homotopic connectivity (VMHC) method was applied to assess the interhemispheric homotopic connectivity changes in patients with CE. Furthermore, the support vector machine method was applied to assess to differentiate patients with CE from healthy controls (HCs) with the VMHC maps as a feature.ResultsCompared with HCs, patients with CE showed significantly increased VMHC values in the bilateral cerebelum_ 8 and cerebelum_4_5. Moreover, we found that the VMHC maps showed an accuracy of 81.94% and an area under the curve of 0.87 for distinguishing the patients with CE from HCs.ConclusionOur study demonstrates that patients with CE showed interhemispheric homotopic connectivity changes in the cerebellum, which might reflect the neurological mechanisms of impaired stereoscopic vision and eye deviation in patients with CE.

Diabetic retinopathy (DR) is the leading cause of decreased vision and blindness globally. The aim of this study was to understand the role of physcion 8-O-β-glucopyranoside (PG) in high glucose (HG)-induced DR and to investigate whether lncRNA NORAD/miR-125/STAT3 signalling was the underlying mechanism involved in DR. To this end, the serum levels of NORAD, miR-125, and STAT3 were determined in patients with DR. The APRE-19 cells were subjected to HG treatment to construct the cell model of DR. HG-disposed APRE-19 cell injury was assessed by detecting cell viability, apoptosis, concentrations of pro-inflammatory cytokines including TNF-α and IL-1β, and ROS generation. Moreover, the effect of PG on HG-disposed APRE-19 cell injury was investigated. NORAD was then overexpressed to investigate the combined effects of NORAD overexpression and PG on HG-disposed APRE-19 cell injury. Furthermore, the regulatory relationship between NORAD and miR-125 as well as miR-125 and STAT3 was investigated. The expression levels of NORAD and STAT3 were significantly increased in the serum of DR patients, while the miR-125 expression was decreased. The HG treatment-induced injury to APRE-19 cells, which were alleviated by PG treatment. Moreover, PG alleviated HG-disposed injury to ARPE-19 cells by decreasing NORAD. NORAD negatively regulated miR-125 expression and the combined effects of NORAD and PG on HG-disposed ARPE-19 cell injury were reversed by miR-125 overexpression. Furthermore, STAT3 was confirmed as a target gene of miR-125. Our results show that PG exerts protective roles in HG-disposed DR

Porcine circovirus 2 (PCV2) causes immunosuppression. Piglets infected with PCV2 can develop enteritis. Given that the gut is the largest immune organ, however, the response of the gut’s immune system to PCV2 is still unclear. Here, IPEC-J2 cells with different treatments were co-cultured with PBMC or CD4+ T cells (Transwell). Flow cytometry and Western blotting revealed that PCV2-infected IPEC-J2 increased the frequency of CD4+ T cells among piglets’ peripheral blood mononuclear cells (PBMCs) and caused CD4+ T cells to undergo a transformation into Foxp3+ regulatory T cells (Treg cells) via activating CD4+ T ERK. Cytokines production and an inhibitor assay showed that the induction of Tregs by PCV2-infected IPEC-J2 was dependent on TGF-β induced by PCV2 in IPEC-J2, which was associated with the activation of NF-κB. Taken together, PCV2-infected IPEC-J2 activated NF-κB to stimulate the synthesis of TGF-β, which enhanced the differentiation of CD4+ T cells into Treg cells through the activation of ERK in CD4+ T cells. This information sheds light on PCV2′s function in the intestinal immune system and suggests a potential immunosuppressive mechanism for PCV2 infection.

Long non-coding RNA (lncRNA) X–inactive specific transcript (XIST) is oncogenic in multiple cancers. Herein, the present study is aimed at delving into how XIST functions in retinoblastoma (RB) and investigating its underlying mechanism. In this study, XIST, miR-191-5p, BDNF mRNA, and BDNF expression levels in RB tissues or cell lines were examined by quantitative real-time polymerase chain reaction (qRT-PCR) or Western blot. The models of gain-of-function and loss-of-function were established by the transfection of pcDNA3.1-XIST, XIST siRNA, and miR-191-5p mimics and inhibitors into SO-Rb50 and Y79 cells, respectively. RB cell proliferation, migration, invasion, and apoptosis were detected employing cell counting kit-8 (CCK-8), Transwell, and terminal deoxynucleotidyl transferased UTP nick end labeling (TUNEL) assays. The regulatory relationships among XIST, miR-191-5p, and BDNF were affirmed utilizing bioinformatics analysis, luciferase reporter assay, qRT-PCR, as well as Western blot. We reported that, XIST expression was markedly elevated in RB tissue and RB cells. XIST overexpression accelerated RB cell proliferation, migration, and invasion, and attenuated RB cell apoptosis but miR-191-5p exerted the opposite effects. Besides, BDNF expression was inhibited by miR-191-5p in both mRNA and protein levels. XIST indirectly improved BDNF expression by repressing miR-191-5p expression as a competitive endogenous RNA. In conclusion, XIST expression is abnormally elevated in RB tissues and XIST can modulate proliferation, migration, invasion, and apoptosis of RB cells by regulating miR-191-5p/BDNF axis., Graphical abstract

Although urbanization has deeply changed farmers’ livelihoods, the improvement of their adaptation to climate change is not yet particularly clear, due to many interactive pressures and complex differences in farmers’ livelihood strategies. This study uses 2-year household survey panel data from Na’Yong and Zhen’Xiong in southwest China, both of which are typically affected by meteorological disasters. First, we constructed a multi-dimensional induction of sensitivity indicators. Then, we divided farmers’ livelihood strategies and identified their sensitivity levels. Finally, we extracted the typical characteristics of farmers’ livelihood strategies that contribute to low sensitivity. The research shows that although non-agricultural labour can improve the income level of farmers, there is no clear positive relationship between farmers’ income and their sensitivity. Levels of sensitivity depend not only on the risk of farmers with a higher exposure to the natural environment but also on the ability of farmers to disperse and avoid natural risks. Livelihood strategies with lower sensitivities have three main characteristics: diversification, marketization and ecological sustainability. This study provides a practical basis for the sustainable livelihood of farmers in the context of climate change and attempts to provide a reference for other developing countries towards the end of achieving ecological poverty alleviation.

Wencui Wan,* Yang Long,* Xuemin Jin, Qiuming Li, Weiwei Wan, Hongzhuo Liu, Yu Zhu Department of Ophthalmology, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan, People’s Republic of China*These authors contributed equally to this workCorrespondence: Yu Zhu Email 13673666718@163.comBackground: Diabetic retinopathy (DR) is a most common microvascular complication and regarded as the leading cause of blindness in the working age population. The involvement of miR-200a in various disorders has become recognized, and the objective of this study was to identify the protective effect of miR-200a in the development of DR.Methods: The contents of miR-200a and its potential target gene, PDZ and LIM domain protein 1 (PDLIM1), were detected in both in-vivo and in-vitro DR models. Retinal leakage and inflammatory factor concentrations were detected after vitreous injections of miR-200a/PDLIM1 vectors in mice. The cellular viability, apoptosis and cellular migration were investigated using trypan blue staining, flow cytometry and transwell assay with human retinal microvascular endothelial cells (HRMECs). Besides, the prediction and confirmation of miR-200a targeting PDLIM1 were conducted with bioinformation analyses and dual-luciferase reporter assay.Results: Lower miR-200a and higher PDLIM1 levels were detected in both in-vivo and in-vitro DR models. Besides, it was found that miR-200a treatment would significantly inhibit retinal permeability and inflammatory factors. Through targeting PDLIM1, it was found that miR-200a could improve cellular viability, remit apoptotic status and reduce cellular migration significantly in high glucose-treated HRMECs.Conclusion: Our results demonstrated that miR-200a could be used as a potential therapy target through down-regulating PDLIM1 in DR.Keywords: microRNA 200a, diabetic retinopathy, PDZ and LIM domain protein 1, endothelial cells

Aim This study aimed to evaluate the effects of Asiatic acid (AA), a naturally occurring compound of pentacyclic triterpenoid, on the pathological processes of diabetic retinopathy (DR). Methods SD rats were induced to develop early DR by intraperitoneal injection of STZ (60 mg/kg). Four weeks after injection, the diabetic rats were orally administrated with 37.5 mg/kg or 75 mg/kg AA every day for four weeks. The integrity of blood-retinal barrier (BRB) was measured by Evans blue staining. The polarization of microglia was determined by real-time PCR, western blot, and ELISA assays. The inner BRB (iBRB) or outer BRB (oBRB) breakdown was induced in human retinal endothelial cells or APRE19 cells through co-culture with high glucose and LPS-stimulated microglia BV2 cells. The damage to the iBRB and oBRB was measured using transendothelial/transepithelial electrical resistance (TEER/TER) and FITC-conjugated dextran cell permeability assays. Key findings Results demonstrated that AA alleviated BRB breakdown, as evidenced by decreased protein expression of occludin, claudin-5, and ZO-1. Furthermore, AA treatment suppressed inflammation and M1 polarization, while it increased M2 polarization in the retina of DR rats. In vitro, the iBRB or oBRB breakdown was alleviated by AA. LPS-induced M1-polarization of BV2 cells under high glucose condition was also repressed through AA administration. Finally, we demonstrated that AA weakened the TLR4/MyD88/NF-κB p65 signaling pathway both in vivo and in vitro. Significance AA ameliorated early DR by regulating microglia polarization via the TLR4/MyD88/NF-κB p65 pathway. These data indicate that AA is a potential candidate for DR treatment.

Retinal vein occlusion (RVO) is the second most common retinal vascular disorders and causes visual damage in a large population. Neutrophil extracellular traps (NETs) formation (NETosis) is an important cause of vascular diseases, however, the association between NETs related biomarkers and RVO development remained unclear. In this pilot study, a total of 77 RVO cases and 48 controls were included between Jan 2020 and July 2020. Besides, the circulating levels of three NETs related markers, cell-free DNA (cfDNA), myeloperoxidase (MPO)-DNA and citrullinated histone H3 (H3Cit), were detected in all the participants and thus the association between NETosis and RVO incidence was analyzed. Advanced assays were conducted to investigate the inflammation and thrombosis related biomarkers in RVO cases with higher or lower NETs biomarkers. When the results were considered, it was found that NETs biomarkers, including cfDNA, MPO-DNA and H3Cit, were increased in the RVO cases comparing with the controls (P 0.05). Through the receiver operating characteristic (ROC) analyses, we found that circulating NETs related biomarkers demonstrated potential diagnostic effects for RVO and the AUCs of plasma cfDNA, MPO-DNA and H3Cit were 0.859, 0.871 and 0.928, respectively (P 0.001). Through analyzing the correlations between circulating NETs markers and RVO stages and durations, inflammatory markers as well as thrombotic indexes, it was found that NETs were related with the RVO subtypes, inflammatory status and thrombus formation. In conclusion, the plasma NETs remnants are significantly increased in RVO cases. Besides, advanced studies demonstrate that inflammation as well as thrombus formation might be involved in this association.

Retinoblastoma is common primary intraocular malignancy of infants and childhood. Neferine is a major bisbenzylisoquinoline alkaloid derived from the lotus plumule in Nelumbo nucifera. This study evaluated the mitigation role of Neferine on retinoblastoma in vitro and in vivo. Xenotransplantation model was established by injecting WERI-Rb-1 cells subcutaneously. Upon induction of retinoblastoma , mice were intraperitoneally injected with Neferine (0, 0.5, 1, 2 mg/kg) or ethanol every 3 days for 30 days. Tumor weight and tumor volume were measured every three days and compared between four groups. Then, mice were sacrificed and immunohistochemical examination was performed to compare Ki67, VEGF content between groups. WERI-Rb-1 cells were used for in vitro experiments and the anti-angiogenic role of Neferine was assessed by analyzing nodes/HPF number. In WERI-Rb-1 xenotransplantation model, compared with control group, 1 mg/kg Neferine treatment significantly inhibited tumor weight (0.39 ± 0.04 g vs. 0.25 ± 0.03 g, P3 vs. 1276 ± 108 mm3, P

Background Enteritis is one of the most frequently reported symptoms in piglets infected with porcine circovirus type 2 (PCV2), but the immunopathogenesis has not been reported. Objectives This study examined the effect of a PCV2 infection on the intestinal mucosal immune function through morphological observations and immune-related molecular detection. Methods Morphological changes within the ileum of piglets during a PCV2 infection were observed. The expression of the related-molecules was analyzed using a gene chip. The immunocyte subsets were analyzed by flow cytometry. The secretory immunoglobulin A (SIgA) content was analyzed by enzyme-linked immunosorbent assay. Results The PCV2 infection caused ileal villus damage, intestinal epithelial cells exfoliation, and an increase in lymphocytes in the lamina propria at 21 days post-infection. Differentially expressed genes occurred in the defense response, inflammatory response, and the complement and coagulation cascade reactions. Most of them were downregulated significantly at the induction site and upregulated at the effector site. The genes associated with SIgA production were downregulated significantly at the induction site. In contrast, the expression of the Toll-like receptor-related genes was upregulated significantly at the effector site. The frequencies of dendritic cells, B cells, and CD8⁺T cells were upregulated at the 2 sites. The SIgA content decreased significantly in the ileal mucosa. Conclusions PCV2 infections can cause damage to the ileum that is associated with changes in immune-related gene expression, immune-related cell subsets, and SIgA production. These findings elucidated the molecular changes in the ileum after a PCV2 infection from the perspective of intestinal mucosal immunity, which provides insights into a further study for PCV2-induced enteritis.

Additional file 2 The transfection efficiency of siRIG-I, siMDA-5 and siMAVS. PIECs were transfected with siRIG-I, siMDA-5, siMAVS or siNC for 24 h. Afterwards, all groups were infected with PCV2 (MOI = 0.5) for 1 h and then cultured for 24 h. The result of qPCR showed the mRNA level of RIG-I, MDA-5 or MAVS at 24 h (A). The result of Western blot showed the protein level of RIG-I (B), MDA-5 (C) or MAVS (D) at 48 h. siNC treatment was used as a negative control and untransfected PIEC was used as a blank control. Data are results of three independent experiments and are represented as the mean and standard deviation (error bars) for each group. **, P

Additional file 1. The original blots for the figures. For our gel data, the target proteins analyzed by western blot were transferred from SDS-PAGE and the PVDF membrane would be sliced into different strips according to their protein size identified with the loading marker. Then the panel of antibodies targeting these proteins were used for western blot analysis. Our SOP for processing the gel data only collected the pictures with target protein regions instead of the whole gel, so here are the original gels for the data we presented in the manuscript.

Background Porcine circovirus (PCV) disease caused by PCV type 2 (PCV2) is mainly attributed to immunosuppression and immune damage. PCV2 can infect vascular endothelial cells and induce high expression of endothelial IL-8. Dendritic cells (DCs), as professional antigen-presenting cells, can not only present antigens but also activate naïve T-cells, causing an immune response. Methods To demonstrate whether endothelial IL-8 is the main factor inhibiting the maturation and related functions of dendritic cells during PCV2 infection, monocyte-derived DCs (MoDCs) and porcine iliac artery endothelial cells (PIECs) processed by different methods were co-cultured in two ways. Flow cytometry, molecular probe labeling, fluorescence quantitative PCR, and the MTS assay were used to detect the changes in related functions and molecules of MoDCs. Results Compared to those in the PIEC-DC group, the endothelial IL-8 upregulation co-culture group showed significantly lower double-positive rates for CD80/86 and MHC-II of MoDCs and significantly increased endocytosis of MoDCs. Meanwhile, the adhesion rate and average fluorescence intensity of MoDCs were significantly downregulated in migration and adhesion experiments. Furthermore, the MHC-I and LAMP7 mRNA levels in MoDCs and the proliferation of MoDC-stimulated T-cells were markedly reduced. However, the changes in MoDCs of the endothelial IL-8 downregulation co-culture group were the opposite. Conclusions PCV2-induced endothelial IL-8 reduces the adhesion and migration ability of MoDCs, resulting in a decreased maturation rate of MoDCs, and further inhibits antigen presentation by DCs. These results may explain the immunosuppressive mechanism of PCV2 from the perspective of the interaction between endothelial cells and DCs in vitro.

Since lncRNAs could modulate neoplastic development by modulating downstream miRNAs and genes, this study was carried out to figure out the synthetic contribution of HOTAIR, miR‐613 and c‐met to viability, apoptosis and proliferation of retinoblastoma cells. Totally 276 retinoblastoma tissues and tumour‐adjacent tissues were collected, and human retinoblastoma cell lines (ie, Y79, HXO‐Rb44, SO‐Rb50 and WERI‐RB1) were also gathered. Moreover, transfections of pcDNA3.1‐HOTAIR, si‐HOTAIR, miR‐613 mimic, miR‐613 inhibitor, pcDNA3.1/c‐met were performed to evaluate the influence of HOTAIR, miR‐613 and c‐met on viability, apoptosis and epithelial‐mesenchymal transition (EMT) of retinoblastoma cells. Dual‐luciferase reporter gene assay was also arranged to confirm the targeted relationship between HOTAIR and miR‐613, as well as between miR‐613 and c‐met. Consequently, up‐regulated HOTAIR and down‐regulated miR‐613 expressions displayed associations with poor survival status of retinoblastoma patients (P < 0.05). Besides, inhibited HOTAIR and promoted miR‐613 elevated E‐cadherin expression, yet decreased Snail and Vimentin expressions (P < 0.05). Simultaneously, cell proliferation and cell viability were also less‐motivated (P < 0.05). Nonetheless, c‐met prohibited the functioning of miR‐613, resulting in promoted cell proliferation and viability, along with inhibited cell apoptosis (P < 0.05). Finally, HOTAIR was verified to directly target miR‐613, and c‐met was the direct target gene of miR‐613 (P < 0.05). In conclusion, the role of lncRNA HOTAIR/miR‐613/c‐met signalling axis in modulating retinoblastoma cells’ viability, apoptosis and expressions of EMT‐specific proteins might provide evidences for developing appropriate diagnostic and treatment strategies for retinoblastoma.

In the present study, a swirl diffuser with a novel design is proposed, in which three geometrical parameters, the angle of the guide vanes φ, the height of the convergent chamber h, and the radius of the diffuser outlet r, can be varied independently. Large eddy simulations were conducted to examine the airflow characteristics downstream of the swirl diffuser. Grid independence was obtained for the grid number 4000,000, and this result was validated by comparison with an experiment. The vorticities, the instantaneous and averaged velocities, and the evolution of the age of air were investigated. The turbulent kinetic energy budget and the momentum balance in the time-averaged Navier–Stokes equations were then studied in an attempt to clarify the mechanism of the swirl diffuser's high diffusing performance. With the increasing swirling strength, the pressure gradient force could not balance the centrifugal force in the radial direction. As a result, the vortex core expanded, generating turbulence. A non-dimensional swirl number parameter was suggested which can be used for determining the structure of the swirl diffuser airflow. To validate this non-dimensional parameter, different combinations of φ, h, and r were examined. It was found that only if the swirl number is the same, will the airflow structure be similar. The results of our study are likely to be useful for obtaining deeper insights about the swirl diffuser airflow, and for better understanding of the effect of the controlling parameter on the swirl diffuser's performance.

The progress of medicine can lead the development of human civilization to a great extent. There is no doubt that the progress of medicine will provide a more happy life for human beings. Nowadays, the number of doctors and nurses in our country is in short supply [1]. As our country is making every effort to develop medical cause, the task of training medical students is very important. The reform of traditional medical education has become the main topic in the medical field. The emergence of computer technology provides the dawn for the future of medical education. With the progress of computer technology, great changes have taken place in the field of medical teaching. This paper briefly introduces the theoretical knowledge of computer teaching. On this basis, this paper introduces the application and influence of computer technology in the field of medical teaching.

Image segmentation is defined by separating scenes into different parts. The grayscale histogram-based method is a widely used method in image segmentation because of its low computational complexity. However, traditional grayscale histogram-based methods usually suffer from wrong segmentation or low structure because of the irregularities and sharp details on the grayscale histogram and the absence of spatial information. This study presents an accurate image segmentation method by using the variational mode decomposition (VMD) and correlation coefficient (CC). First, grayscale histogram is decomposed into certain band-limit modes by VMD to remove the adverse effects. Then, the candidate centroids of the clusters are obtained by searching the refined histogram both vertically and horizontally. The K-means method is used to generate the segmentation results. Finally, some over-segmented regions are merged following the CCs of spatial histogram, which is also refined by the VMD method. The proposed method was applied to four images. The obtained experimental results show that the proposed method can generate reasonable segmentation results than those of the existing image segmentation algorithms.

Background/Aims: Age-related macular degeneration (AMD) is the primary cause of senior blindness in developed countries. Mechanisms underlying initiation and development of AMD remained known. Methods: We examined the CD4+ T cell compartments and their functions in AMD patients. Results: AMD patients presented significantly higher frequencies of interferon (IFN)-γ-expressing and interleukin (IL)-17-expressing CD4+ T cells than healthy controls. The levels of IFN-γ and IL-17 expression by CD4+ T cells were significantly higher in AMD patients. These IFN-γ-expressing Th1 cells and IL-17-expressing Th17 cells could be selectively enriched by surface CCR3+ and CCR4+CCR6+ expression, respectively. Th1 and Th17 cells from AMD patients promoted the differentiation of monocytes toward M1 macrophages, which were previously associated with retinal damage. Th1 and Th17 cells also increased the level of MHC class I expression in human retinal pigment epithelial (RPE)-1 cells, while Th1 cells increased the frequency of MHC class II-expressing RPE-1 cells. These proinflammatory effects were partly, but not entirely, induced by the secretion of IFN-γ and IL-17. Conclusions: This study demonstrated an enrichment of Th1 cells and Th17 cells in AMD patients. These Th1 and Th17 cells possessed proinflammatory roles in an IFN-γ- and IL-17-dependent fashion, and could potentially serve as therapeutic targets.

BACKGROUND: The present study aimed to investigate the protective role of leukemia inhibitory factor (LIF) against oxidative damage in photoreceptor cone cells. METHODS: In vivo, dark-adapted mice were injected with LIF or phosphate-buffered saline (PBS) intravitreously prior to being exposed to 5,000 lux bright light to determine the protective effect of LIF against light damage in cone cells. Oxidative damage to cone cells was analyzed using electroretinograms, immunostaining, Western blotting and reverse transcription quantitative polymerase chain reaction (RT-qPCR). In vitro, 661W cells were pretreated with 5 ng/mL of LIF with or without 50 µM of signal transducer and activator of transcription 3 (STAT3) inhibitor S3I201 for 1 h prior to treatment with 1 mM H(2)O(2); cell survival, apoptosis, the oxidative stress index, and the activation of STAT3, extracellular signal-regulated kinase (ERK1/2), and AKT were subsequently determined. RESULTS: In vivo, light induction damaged the function and morphology of cone cells, and LIF was observed to protect cone cells from this light damage. Moreover, the activation of the Janus tyrosine kinase (JAK)/STAT3 signaling pathway and the subsequent changes in apoptosis and proliferation-related genes were found to be involved in the protective effect of LIF against light-induced retinal damage. In the H(2)O(2)-induced 661W cell model, H(2)O(2) increased cellular apoptosis rates, the expression levels of Bcl-2–associated X-protein (BAX) and cleaved caspase 3, reactive oxygen species (ROS) production, and malondialdehyde content, while decreasing the cell viability, and Bcl-2, superoxide dismutase, catalase, and glutathione peroxidase activity. LIF was observed to block these events; however, the administration of the STAT3 inhibitor S3I201 reversed the beneficial effects of LIF on H(2)O(2)-triggered apoptosis and ROS production. CONCLUSIONS: In conclusion, the present study suggested that LIF may relieve oxidative damage in cone cells through suppressing apoptosis and oxidative stress by targeting the STAT3 signaling pathway.

Over the last two decades, extensive literature has examined the socioeconomic and environmental impacts of China's Sloping Land Conversion Program (SLCP), a program that was launched in late 1990s to mitigate the environmental effects of agricultural production and reduce rural poverty. However, little empirical evidence exists with regard to the impact of SLCP on rural households' sensitivity to nature-induced changes and environmental challenges. In this study, household-level data covering the period 1995–2010 from five Chinese provinces were used to examine the effect of SLCP on farmers' sensitivity to climate change. The empirical results show that participation in SLCP significantly reduced farmers' sensitivity to climate change by reducing their dependency on land and natural resources for income, and by diversifying their livelihood options. Spatially, the results reveal that the effect of SLCP on farmers' sensitivity vary across regions. Specifically, SLCP was found to have a ‘rate effect’ on farmers in the Northern regions and a ‘level effect’ on farmers in the Southern regions. Likewise, we found that the effect of SLCP differs considerably across income groups, with the effect on low- and middle-income groups being most significant. The results indicate that subsidy is the main pathway through which SLCP reduces farmers' sensitivity to climate change. In contrast, we found inclusive evidence about the indirect effect of SLCP farmers' sensitivity through the promotion of non-agricultural employment. These results carry major implications with regard to the effectiveness of ecological conservation programs and their mitigation potential through building farmers' resilience in China and ecologically fragile environments.

Disease biomarkers for diagnostic and prognostic purposes are most likely within an extremely low concentration range and are thus masked by the presence of high‑abundance proteins. Therefore, removing high‑abundance proteins is the main challenge for identifying disease biomarkers. In addition, the solution obtained from high‑abundance protein depletion kits contains a rich array of compounds, which interfere with isoelectric focusing (IEF). In the present study, the effect of two commercial kits was evaluated and the downstream IEF protocol was optimized. High‑resolution results could be obtained according to the following conditions: The ProteoPrep Blue Albumin and IgG Depletion kit depleted albumin and IgG; immobilized pH gradient strips (typically 18 cm) were rehydrated with sample buffer containing 250 µg serum proteins at 30 v for 6 h, 60 v for 6 h, 200 v for 2 h, 500 v for 2 h, 1,000 v for 2 h, 5,000 v for 2 h, 10,000 v for 2 h and then focusing at 10,000 v up to 110 k vhs. In addition, the protein spots identified by matrix‑assisted laser desorption ionization time‑of‑flight mass spectrometry demonstrated that all proteins had a low abundance. The present study not only provides a definite and effective method for removing high‑abundance proteins, but also provides a proper protocol (protocol C) for downstream IEF. The present study includes a comprehensive investigation of serum proteomics, which paves the way for serum protein research.

Aberrant expression of microRNAs plays an important role in the pathogenesis and progression of retinoblastoma. MiR-25, a member of the miR-106b˜25 cluster, has been reported to be abnormally expressed in retinoblastoma, but the exact role of it remains unclear. In our study, we found that miR-25-3p was upregulated in retinoblastoma tissues and cell lines. Enforced expression of miR-25-3p in retinoblastoma cell line WERI-RB-1 increased cell growth, colony formation, anchorage-independent growth, cell migration and invasion in vitro and tumor xenograft growth in vivo. In contrast, inhibited miR-25-3p expression in retinoblastoma cell line Y79 suppressed cell growth, colony formation, anchorage-independent growth, cell migration and invasion. Through luciferase reporter assay, we found that phosphatase and tensin homolog (PTEN) was a direct target of miR-25-3p. This was verified by western blot that miR-25-3p overexpression suppressed PTEN and activated Akt signaling. In addition, miR-25-3p was found to promote epithelial-mesenchymal transition (EMT) of WERI-RB-1 cells through PTEN/Akt pathway. Western blot analysis revealed that miR-25-3p overexpression increased Vimentin and Snail expression, and suppressed E-cadherin expression, but this could be reversed by restoring PTEN. Moreover, LY294002 treatment or restoring PTEN expression abolished the effects of miR-25-3p on cell invasion, colony formation and anchorage-independent growth in vitro and tumor xenograft growth in vivo. Taken together, our results suggested that miR-25-3p promotes malignant transformation of retinoblastoma cells by suppressing PTEN.

Purpose. To investigate the efficacy and safety of combined vitrectomy with tumor resection in the treatment of retinal vasoproliferative tumors (RVPT). Methods. Retrospective study. RVPT patients who underwent vitreous surgery at the Eye Hospital of Wenzhou Medical University from January 2011 to July 2017 were included. The main outcomes included treatment type, tumor activity, and best-corrected visual acuity (BCVA). Results. Altogether, 16 patients with 17 eyes were enrolled with follow-up of no less than 6 months. Eight eyes were in the resection treatment group (Group R) and 9 eyes were in the conservative treatment group (Group C). Female (69%) were more common. The mean age was 50 (49.72 ± 12.92) years. Fifteen patients got unilateral onset and only one patient suffered bilaterally. The common symptoms were decreased visual acuity, floaters, and visual distortion. The preoperative BCVA ranged from hand movement to 20/20, with an average of 0.82 ± 0.75 LogMAR. Patients were all not high myopia, with a mean axial length of 23.27 ± 0.27 mm (21.61 mm to 24.67 mm). Of the retinal diseases, the epiretinal membrane was the most common, followed by vitreous hemorrhage, uveitis, subretinal fluid, and so on. Compared with the baseline BCVA, it improved more at postoperative 6 months and the last visit in Group R than in Group C (P=0.006 and P=0.033). The BCVA-improved 0.2 LogMAR or above in 6 months was 2 eyes in Group C and 7 eyes in Group R. All tumors in Group R were completely resected, whereas three in Group C (33.3%) had definite activity (P=0.008). In all samples, tumors were located on the inner side of the retina and had small vessel wall thickening and hyaline degeneration. The degree of astrocyte proliferation varies widely among different tumors. Conclusions. RVPT was more likely to occur in nonhigh myopia patients. Epiretinal membrane and vitreous hemorrhage were the main causes for vitreous surgery in RVPT patients. Compared with conservative treatment, surgical resection of the tumor is more beneficial to patients on visual acuity recovery and preventing tumor relapse. It is a safe and effective way to treat RVPT.

Proliferative vitreoretinopathy (PVR) is the leading cause of retinal detachment failure. The mechanism of PVR development is complex and still not completely elucidated. There are no proven methods for early prevention or clinical treatment. Retinal proteins are abnormally expressed during the entire PVR disease process. Due to the limitations of research methods and techniques, we do not fully understand the retinal protein changes in PVR. This proteomics study systemically analyzed and identified differential protein expression between retinas of PVR and non-PVR (normal) eyes. Retinal samples were subjected to sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) coupled with mass spectrometry. Raw data were processed and analyzed by Maxquant software and then searched against the human UniProKB (201510) protein database. Differentially expressed proteins were selected and further validated in a human retinal pigment epithelial (RPE) cell line. The effects of dysregulated proteins on cell proliferation, apoptosis, and migration were studied. Systemic proteomics analysis identified several PVR-enriched proteins. The differentially expressed proteins were analyzed by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) annotation to find abnormal pathways involved in PVR. Retinal-specific ATP-binding cassette transporter (ABCA4) expression was one of the most increased proteins in PVR tissue. ABCA4 knockdown significantly reduced proliferation and affected the cell cycle in the human RPE cell line. ABCA4 knockdown also induced apoptosis and inhibited retinal cell migration. In conclusion, systemic proteomics analysis identified differentially expressed proteins in traumatic PVR, with ABCA4 being highly expressed. Disruption of ABCA4 expression induced apoptosis and inhibited cell proliferation and migration in a human RPE cell line.

The purpose of this investigation was to explore the combined effects of single nucleotide polymorphisms (SNPs) within LFA-1/ICAM-1/GSK-3β pathway and environmental hazards on susceptibility to Graves' opthalmopathy (GO) among a Chinese Han population. Altogether 305 GO patients and 283 Graves' disease (GD) subjects were recruited. Information relevant to the participants' age, gender, body mass index (BMI), regular physical activity, smoking history, alcohol intake, stressful work environment, stress at work, family history of thyroid disease and 131I treatment were summarized, and the participants' related SNPs of LFA-1/ICAM-1/GSK-3β were also detected. Then the gene-gene and gene-environment interactions were evaluated by logistic regression model and multi-factor dimensionality reduction (MDR) modeling. The results exhibited that age, BMI, smoking history, stressful work, stress at home, family history of thyroid disease and 131I treatment appeared as potential indicators regulating GO risk, when either univariate or multivariate regression analysis was performed (all P C) and rs2230433 (G > C) of LFA-1, rs1799969 (G > A) and rs5498 (A > G) of ICAM-1, as well as rs6438552 (T > C) and rs334558 (T > C) of GSK-3β were significantly associated with altered susceptibility to GO under the allelic models (all P < 0.05). Also haplotype TGAATC acted as a protective factor against GO risk (P < 0.05), whereas haplotype CGAACC largely elevated risk of GO (P < 0.05). Besides, logistic regression analysis demonstrated that rs12716927, rs5498 and rs6438552 all would affect the influences exerted by age, BMI, smoking history, stressful work, stress at home, family history of thyroid disease or 131I treatment on GO susceptibility (all P < 0.05). MDR modeling implied that the combined model of rs12716977, rs2230433 and rs1799969 was the supreme interactive model when BMI was co-assessed, and the interactive model of rs12716977, rs334558 and rs5491 was the most desirable among the smoking population. In conclusion, gene-gene and gene-environment interactions served as a crucial manner in affecting susceptibility to GO, providing solid evidences for screening effective GO-susceptible biomarkers and exploring potential GO treatment strategies.

The present study aimed to explore the combined role of microRNA (miR)-34a, melanoma antigen-A (MAGE‑A) and p53 in altering the chemosensitivity of retinoblastoma (RB) cells. Human RB and adjacent tumor tissues, as well as human RB cell lines (HXO‑Rb44, SO‑Rb50, Y79 and WERI‑Rb-1) were used. In addition, four chemotherapeutic drugs, including carboplatin, etoposide, Adriamycin and vincristine, were used to treat the cell lines, in order to evaluate the sensitivity of RB cells. Furthermore, miR‑34a expression was detected by reverse transcription-quantitative polymerase chain reaction, and western blotting was implemented to quantify expression levels of MAGE‑A and p53. A luciferase reporter gene assay was used to validate the targeted association between miR‑34a and MAGE‑A. The results indicated that SO‑Rb50 cells exhibited the highest resistance to carboplatin, Adriamycin and vincristine (P

Background Genetic and environmental factors both play important roles in the occurrence and progression of diabetic retinopathy (DR). IL-10 592 gene polymorphism is associated with diabetes pathogenesis. This study analyzed the relationship between IL-10 gene promoter-592 loci polymorphism (SNP) in a diabetic model rats with DR. Material/Methods Streptozotocin (STZ) was injected through the tail vein to establish a diabetic rat model. The rats were randomly divided into 2 groups for 3 months’ feeding, including 100 rats in the diabetes-positive control group and 100 rats only injected with citric acid buffer as the blank control group. Fundus fluorescein angiography (FFA) was used to observe retinal vascular changes. Polymerase chain reaction-restriction fragment polymorphisms assay (PCR-RFLP) was used to detect IL-10 gene promoter-592 loci polymorphism in DNA samples. Enzyme-linked immunosorbent assay (ELISA) was performed to test serum IL-10 concentration. Results Serum IL-10 level in DR rats was 33.18±5.0 pg/mL and in the control rats it was 53.33±4.16 pg/mL in (P

To investigate the effect and safety of vitreous liquefaction induced by C3F8 (an inert gas) injected into vitreous cavit of rabbit eyes. 24 rabbits (48 eyes) were randomly divided into four groups, named group A, group B, group C and group D, with 6 rabbits in each group. The right eye in each rabbit was taken as the experimental eye while the left as the control eye. The experimental eyes in group A were injected with 0.1mL disinfectant air; the experimental eyes in group B, group C and group D were all injected with C3F8 0.1mL, 0.2mL and 0.3mL respectively after receiving anterior chamber penetration; and the controlled eyes in all group were injected with 0.1mL balanced salt solution (BSS). During the first 7 d after injection, all the rabitts' eyes were examined by slit lamp, ophthalmoscope, intraocular pressure (IOP) and dark-adapted retina Electroretinography (ERG) each day. After that, the examination of IOG and ERP were reviewed weekly. Besides, B ultrasound should be examined to observe the situation of posterior vitreous detachment (PVD) in the 4th and 8th weeks. The rabbits were killed in the end of the 8th week, with their specimens examined by the light microscope, scanning electron microscopy (SEM) and transmission electron microscopy (TEM). Except group A, all the experimental eyes were produced with vitreous liquefaction. In group C and group D, in addition to the produced vitreous liquefaction, posterior vitreous detachment (PVD), even complete PVD, were induced in different extent. But in group B, the vitreous body was returned to the gel state at 2 weeks after gas absorption. In group C and group D, the vitreous body was not found to recover its original state at 8 weeks. In group D, there was a little increase of intraocular pressure, a mild delay of wave a and wave b after ERG in the 4th day after the gas injection. While there was no such situation in other groups. After the examination of B ultrasound in the 8th week, the complete PVD occurred in group C (with 2 experimental eyes occurred) and group D (with 4 experimental eyes occurred). Light microscope and transmission electron microscope examination showed no obvious abnormality. Smooth inner limiting membrane could be seen in the eye with PVD occurred when scanning electron microscope used. The injection of C3F8 into rabbit eyes can improve the vitreous liquefaction of the vitreous body and a certain volume of C

This paper proposes a full-order nonsingular terminal sliding mode control method for a variable-speed wind turbine operating in the above-rated regime considering the model uncertainty caused by the constantly changing operating point and external disturbances. The proposed control method can not only converge to the equilibrium point in finite time, but also overcome the model uncertainty and guarantee the stability of the control system. The proposed control strategy is designed to minimize rotor speed error. A group of actual data obtained from a wind farm is used in the simulation study, the result shows that the proposed method can maintain the rotor speed at the rated value and keep the output power stable. In order to assess the effectiveness of the proposed method, the paper compares the proposed method with traditional terminal sliding mode control. The results confirm that the proposed method is fast, robust, and effective.

MicroRNAs are increasingly recognized as important regulators of cancer. The aim of the present study was to investigate the role of miR-106b in the regulation of Y79 retinoblastoma. Y79 cells were transfected with antisense oligonucleotides (ASO) against miR-106b (ASO-miR-106b) or ASO-control. After transfection, the levels of miR-106b were monitored with real-time PCR (RT-PCR). The effects of ASO-miR-106b transfection on cell viability was evaluated by Cell Counting Kit-8 (CCK-8) analysis at 24, 48 and 72 h after transfection. Subsequently, the cells were stained with Annexin V-FITC and propidium iodide (PI) and subjected to flow cytometry to assess cell apoptosis. Transwell assay was used to analyze cell migration. Changes in Runt-related transcription factor 3 (Runx3) mRNA and proteins levels were also evaluated. miR-106b was downregulated by ASO-miR-106b at 48 and 72 h after transfection, accompanied by a decrease in cell viability and proliferation, as well as an increase in cell apoptosis. Transwell analysis indicated that cells treated with ASO-miR-106b exhibited significantly lower cell migratory abilities. The mRNA and protein level of Runx3 were upregulated after transfection. These results demonstrated that suppression of miR-106b inhibited Y79 cell proliferation and migration. The upregulation of Runx3 after miR-106b suppression ascertained that Runx3 is a tumor-suppressor in retinoblastoma and is a target of miR-106b.

This study aimed to investigate the correlation between age-related macular degeneration (AMD) of the liver-kidney yin-deficiency type and complement factor H (CFH) polymorphism, and to determine whether the C allele of the T1277C (Y402H) variant is a risk factor for this condition. We performed a case-control investigation of 60 patients with liver-kidney yin-deficiency AMD and 60 normal control subjects. Peripheral blood was collected from each participant for DNA extraction. Following amplification by polymerase chain reaction, the DNA samples were sequenced, and polymorphism of the CFH gene was examined. Data were analyzed with the chi-square test, with P < 0.05 signifying statistical significance. The frequency of the C allele was significantly higher in the wet than in the dry AMD group (P = 0.044). In addition, the TC and CC genotypes were markedly more common in the former than in the control group (P = 0.013), and there was a significant difference in the distribution of the T and C alleles between wet AMD patients and control subjects (P < 0.05). Based on this, we conclude that liver-kidney yin-deficiency AMD is associated with the C allele and TC and CC genotypes of the CFH Y402H polymorphism. Among patients with this condition, CFH genotypes were normally distributed. The principal CFH genotypes that induce liver-kidney yin-deficiency AMD are the mutant homozygote CC and heterozygote TC forms. Moreover, C allele carriers are at higher risk of developing this disease.

The linearly constrained least squares constant modulus algorithm (LSCMA) may suffer significant performance degradation and lack robustness in the presence of the slight mismatches between the actual and assumed signal steering vectors, which can cause the serious problem of desired signal cancellation. To account for the mismatches, we propose a doubly constrained robust LSCMA based on explicit modeling of uncertainty in the desired signal array response and data covariance matrix, which provides robustness against pointing errors and random perturbations in detector parameters. Our algorithm optimizes the worst-case performance by minimizing the output SINR while maintaining a distortionless response for the worst-case signal steering vector. The weight vector can be optimized by the partial Taylor-series expansion and Lagrange multiplier method, and the optimal value of the Lagrange multiplier is iteratively derived based on the known level of uncertainty in the signal DOA. The proposed implementation based on iterative minimization eliminates the covariance matrix inversion estimation at a comparable cost with that of the existing LSCMA. We present a theoretical analysis of our proposed algorithm in terms of convergence, SINR performance, array beampattern gain, and complexity cost in the presence of random steering vector mismatches. In contrast to the linearly constrained LSCMA, the proposed algorithm provides excellent robustness against the signal steering vector mismatches, yields improved signal capture performance, has superior performance on SINR improvement, and enhances the array system performance under random perturbations in sensor parameters. The on-line implementation and significant SINR enhancement support the practicability of the proposed algorithm. The numerical experiments have been carried out to demonstrate the superiority of the proposed algorithm on beampattern control and output SINR enhancement compared with linearly constrained LSCMA.

Based on 1-D fractional Fourier transform, we proposed an image encryption algorithm in order to hide two images simultaneously. When the fractional order is closed to 1, most energy in frequency domain is centralized in the center part of spectrum. The image can be recovered acceptable by using a half of spectrum, which locates in the middle part at x-direction or y-direction. Cutting operation is employed in order to combine two spectra. Double random phase encoding is employed for image encryption. The corresponding numerical simulations are performed to demonstrate the validity and efficiency of the algorithm.

Infrared images are firstly analyzed using the multifractal theory so that the singularity of each pixel can be extracted from the images. The multifractal spectrum is then estimated, which can reflect overall characteristic of an infrared image. Thus the edge and texture of an infrared image can be accurately extracted based on the singularity of each pixel and the multifractal spectrum. Finally the edge pixels are classified and enhanced in accordance with the sensitivity of human visual system to the edge profile of an infrared image. The experimental results obtained by this approach are compared with those obtained by other methods. It is found that the proposed approach can be used to highlight the edge area of an infrared image to make an infrared image more suitable for observation by human eyes.

Alpha-thalassemia (thal) is one of the most common genetic diseases and is widely distributed in southern China. Fetuses with homozygous alpha(0)-thal, known as Hb Bart's (gamma(4)) disease, usually die during the last trimester of gestation, and the affected pregnancies are frequently associated with serious maternal morbidity and mortality. Prenatal diagnosis is needed for early detection of this disease in couples at-risk. This can be performed by invasive fetal sampling or by serial ultrasonography to detect affected fetuses followed by invasive testing. Of 122 at-risk families who received prenatal testing, 98 (80.3%) chose ultrasonography, and 24 (19.7%) chose direct invasive testing. In all, 28 affected pregnancies were detected and terminated. Ultrasonography is now playing a major role in prenatal control of Hb Bart's disease in our region, and limits the invasive procedures to the few patients identified to be at high risk.

In this paper, an airdropping and unfolding scheme for tethered balloon platform is proposed firstly. And the details of the system structure is design, the key cushioning pad honeycomb paperboard is studied. Based on the designed structure, the cushioning simulation is conducted and the results show that the cushioning performance is feasible for airdrop application.

Transmission and reflection properties of a Gaussian beam incident upon a dielectric slab are investigated by means the plane-wave spectrum method and compared with experimental results made at 110 GHz. The incident wave generated by a special circular horn antenna at 110 GHz is modeled as a two-dimensional Gaussian beam under the paraxial approximation and also as a simplified one-dimensional beam with a spreading factor. Measurement results are presented for the transmission and reflection of both the perpendicular and parallel polarizations. In particular, reflection measurements for a parallel-polarized beam incident at and near the Brewster angle are shown and compared to the theory. A small reflected two-lobe beam is observed even at the exact Brewster angle. Nonspecular phenomena in Gaussian beams are discussed and the relevance to a Brewster window for the output of a gyrotron is considered