Fasudil-modified macrophages reduce inflammation and regulate the immune response in experimental autoimmune encephalomyelitis

Multiple sclerosis is characterized by demyelination and neuronal loss caused by inflammatory cell activation and infiltration into the central nervous system.Macrophage polarization plays an important role in the pathogenesis of experimental autoimmune encephalomyelitis,a traditional experimental model of multiple sclerosis.This study investigated the effect of Fasudil on macrophages and examined the therapeutic potential of Fasudil-modified macrophages in experimental autoimmune encephalomyelitis.We found that Fasudil induced the conversion of macrophages from the pro-inflammatory M1 type to the anti-inflammatory M2 type,as shown by reduced expression of inducible nitric oxide synthase/nitric oxide,interleukin-12,and CD16/32 and increased expression of arginase-1,interleukin-10,CD14,and CD206,which was linked to inhibition of Rho kinase activity,decreased expression of toll-like receptors,nuclear factor-κB,and components of the mitogen-activated protein kinase signaling pathway,and generation of the pro-inflammatory cytokines tumor necrosis factor-α,interleukin-1β,and interleukin-6.Crucially,Fasudil-modified macrophages effectively decreased the impact of experimental autoimmune encephalomyelitis,resulting in later onset of disease,lower symptom scores,less weight loss,and reduced demyelination compared with unmodified macrophages.In addition,Fasudil-modified macrophages decreased interleukin-17 expression on CD4^(+)T cells and CD16/32,inducible nitric oxide synthase,and interleukin-12 expression on F4/80^(+)macrophages,as well as increasing interleukin-10 expression on CD4^(+)T cells and arginase-1,CD206,and interleukin-10 expression on F4/80^(+)macrophages,which improved immune regulation and reduced inflammation.These findings suggest that Fasudil-modified macrophages may help treat experimental autoimmune encephalomyelitis by inducing M2 macrophage polarization and inhibiting the inflammatory response,thereby providing new insight into cell immunotherapy for multiple sclerosis.

A Local-Global Attention Fusion Framework With Tensor Decomposition for Medical Diagnosis

Dear Editor,In this letter,a novel hierarchical fusion framework is proposed to address the imperfect data property in complex medical image analysis(MIA)scenes.In particular,by combining the strengths of convolutional neural networks(CNNs)and transformers,the enhanced feature extraction,spatial modeling,and sequential context learning are realized to provide comprehensive insights on the complex data patterns.Integration of information in different level is enabled via a multi-attention fusion mechanism,and the tensor decomposition methods are adopted so that compact and distinctive representation of the underlying and high-dimensional medical image features can be accomplished[1].It is shown from the evaluation results that the proposed framework is competitive and superior as compared with some other advanced algorithms,which effectively handles the imperfect property of inter-class similarity and intra-class differences in diseases,and meanwhile,the model complexity is reduced within an acceptable level,which benefits the deployment in clinic practice.MIA has assumed a pivotal role in numerous critical clinical scenarios,where sophisticated image analysis techniques have proven instrumental in augmenting medical decision-making,facilitating individualized therapeutic interventions,and enhancing patient prognostication[2]−[4].

SiRAP2-12,a Positive Regulatory Factor,Effectively Improves the Waterlogging Tolerance of Foxtail Millet(Setaria italica)

Foxtail millet(Setaria italica)growth was inhibited because of waterlogging stress,which has caused yield reduc-tion.ERF family plays an important role to plant adversity tolerance.In our study,we obtained 19,819 differential expressed genes(DEGs)between the two treatments based on the RNA-seq sequencing of foxtail millet of water-logging stress.Furthermore,a total of 28 ERF family members were obtained,which have a complete open read-ing frame.We studied the evolution and function of SiERF family and how they affected the waterlogging tolerance.It was found that SiERF1A/B/C(GenBank ID:OR775217,OR775219,OR775218)and SiRAP2-12(GenBank ID:OR775216)have similar functions to the known waterlogging tolerance genes of other plants.Among them,the SiRAP2-12 expression was obviously significantly up-regulated in foxtail millet after 5d water-logging stress.After SiRAP2-12 was silenced,the activity of defense enzymes in millet decreased significantly.In details,superoxide dismutase(SOD),catalase(CAT)and peroxidase(POD),the osmotic regulator proline(Pro),and the activity of the anaerobic respiratory enzyme alcohol dehydrogenase(ADH)content were decreased by 78.61%,29.52%,79.95%,19.41%and 54.77%,respectively.In contrast,the relative electrical conductivity contents(REC),malondialdehyde(MDA),and hydrogen peroxide(H_(2)O_(2))of the foxtail millet subjected to virus-induced gene silencing clearly increased by 1.03-fold,36.09%,and 15.21%,respectively.The content of sodium(Na^(+))in the SiRAP2-12-silenced foxtail millet also increased,but that of potassium(K^(+))decreased.Interestingly,we found that ethylene content was significantly reduced.Further,the SiAOC1 expression,an essential gene for ethylene synthesis,was inhibited in SiRAP2-12-silenced foxtail millet after waterlogging stress.Taken together,we hypothesized that SiRAP2-12 might be a positive regulator of millet tolerance to waterlogging stress.

Expression Pattern, Interaction Network, and Functional Analysis of the Arabidopsis Botrytis Susceptible1 Interactor

E3 ubiquitin ligases are participated in numerous processes, regulating the response to biotic and abiotic stresses. Botrytis susceptible1 interactor (BOI) is a RING (Really Interesting New Gene)-type E3 ligase that mediates the ubiquitination of BOS1 (Botrytis susceptible1), a transcription factor involved in stress and pathogen responses. Although BOI is an E3 ligase, there are reports to show that BOI interacts with target proteins such as DELLAs or CONSTANS to repress gibberellin responses and flowering without the degradation of the target proteins. In this article, we utilize diversified methods to comprehensively analyze the expression pattern, interaction network and function of BOI gene. Firstly, 1800 bp upstream region of BOI gene from Arabidopsis thaliana (Arabidopsis) genome was isolated, and fused GUS reporter gene. The resulting expression cassette was introduced into wild-type Arabidopsis through Agrobacterium-mediated transformation. The result demonstrated that BOI gene was expressed predominantly in leaves, siliques, young roots, and flowering tissues, indicating that BOI gene may be involved in multiple processes in plant growth and development in Arabidopsis. Besides, eight candidate interacting proteins were obtained from the Arabidopsis cDNA library via yeast two-hybrid technology, including EXO70E2 (AT5G61010), WRKY7 (AT4G24240), WRKY11 (AT4G31550), WRKY17 (AT2G24570), UBP20 (AT4G17895), L5 (AT1G12290), SAUR9 (AT4G36110) and TCP21 (AT5G08330). Functional analysis of these candidate interacting proteins manifested that they related to multiple pathways, including biological and abiotic stress, programmed cell death, protein degradation, material metabolism and transcriptional regulation. In addition, the results of the transient assay proclaimed that BOI protein affects the protein stability of EXO70E2 and L5 through its E3 ubiquitin ligase activity. Our results provide novel clues for a better understanding of molecular mechanisms underlying BOI-mediated regulations.

Dissecting Multiple Arabidopsis CC-NBS-LRR Proteins Structure and Localization

NBS-LRR (nucleotide binding sites and leucine rich repeat) protein plays a crucial role as sentries and as defense activators in plants. The structure and function of NBS-LRR proteins are closely related. Previous articles have announced that the activated ZAR1 (HopZ-Activated Resistance 1) forms a pentamer in the plasma membrane, which is a calcium permeable channel that can trigger plant immune signaling and cell death. However, the structure of galore NBS-LRRs in Arabidopsis is not yet clear. The functional sites of distinct NBS-LRR in cells may vary. In addition, identifying pathogens and activating defense regions may occur in different subcellular compartments. Therefore, dissecting the specific structure and positioning of NBS-LRRs is an indispensable step in understanding their functions. In this article, we exploit AlphaFold to predict the structure of some designed NBS-LRRs, and utilize Agroinfiltration transient expression system, combined with biochemical fractionation, to dissect the localization of these NBS-LRR receptors from Arabidopsis. Structural data indicates that the identified NBS-LRRs share analogous conformation. Membrane fractionation assay demonstrates these NBS-LRRs are mainly associated with the membrane. These data show that the Ca2+-permeable channel activity may be evolutionarily conserved in NBS-LRR of Arabidopsis, and this study provides some reference clues for analyzing the structure and localization patterns of other plant immune receptors.

Trends in research related to high myopia from 2010 to 2019:a bibliometric and knowledge mapping analysis

AIM:To evaluate the global trends in and explore hotspots of high myopia(HM)research.METHODS:This bibliometric analysis was used to reveal the publication trends in HM research field based on the Web of Science Core Collection(WoSCC).VOSviewer version 1.6.13 software was used to analyze the data and construct a knowledge map including the yearly publication number,journals,countries,international collaborations,authors,research hotspots,and intellectual base in HM.RESULTS:The search engine found 3544 peer-reviewed publications on HM between 2010 and 2019,and the yearly research output substantially elevated over the past decade.China is the top publishing country,and Sun Yatsen University was the most active academic institution.Jonas JB is the top publishing scientist,and Investigative Ophthalmology and Visual Science(IOVS)was the most productive journal.The highest cited references mainly focused on epidemiology and management.The keywords formed 6 clusters:1)refractive surgery;2)etiology and clinical characteristics;3)the mechanism of eye growth;4)management for myopic maculopathy;5)vitrectomy surgical treatment;6)myopia-associated glaucoma-like optic neuropathy.CONCLUSION:The evaluation of development trends based on the data extracted from WoSCC can provide valuable information and guidance for ophthalmologists and public health researchers to improve management procedures in HM field.

The effects and potential of microglial polarization and crosstalk with other cells of the central nervous system in the treatment of Alzheimer’s disease

Microglia are resident immune cells in the central nervous system. During the pathogenesis of Alzheimer’s disease, stimulatory factors continuously act on the microglia causing abnormal activation and unbalanced phenotypic changes;these events have become a significant and promising area of research. In this review, we summarize the effects of microglial polarization and crosstalk with other cells in the central nervous system in the treatment of Alzheimer’s disease. Our literature search found that phenotypic changes occur continuously in Alzheimer’s disease and that microglia exhibit extensive crosstalk with astrocytes, oligodendrocytes, neurons, and penetrated peripheral innate immune cells via specific signaling pathways and cytokines. Collectively, unlike previous efforts to modulate microglial phenotypes at a single level, targeting the phenotypes of microglia and the crosstalk with other cells in the central nervous system may be more effective in reducing inflammation in the central nervous system in Alzheimer’s disease. This would establish a theoretical basis for reducing neuronal death from central nervous system inflammation and provide an appropriate environment to promote neuronal regeneration in the treatment of Alzheimer’s disease.

Endoscopic transnasal optic canal decompression for pediatric traumatic optic neuropathy with no light perception

Dear Editor,We reported the results of endoscopic transnasal optic canal decompression(ETOCD)procedure in 3 pediatric cases(9 to 12y,mean age 10.3y)of traumatic optic neuropathy(TON)with no light perception suffering from road traffic accident.TON is a rare but serious complication secondary to ocular or head trauma,resulting in partial or complete visual loss.In China,the incidence of road traffic accident induced TON is on the rise.The rate of incidence has been reported about 0.5% to 5% in closed head trauma cases[1].

Comparison of the Histological Morphology between Normal Skin and Scar Tissue

Skin wound healing is a complex event, and interrupted wound healing process could lead to scar formation. The aim of this study was to examine the morphological changes of scar tissue. Pathological staining（HE staining, Masson＇s trichrome staining, methenamine silver staining） was used to evaluate the morphological changes of regenerating epidermis in normal skin and scar tissue, and immunofluorescence staining to detect the expression of collagen Ⅳ, a component of basement membrane（BM）, and the expression of integrinβ4, a receptor for BM laminins. Additionally, the expression of CK14, CK5, and CK10 was measured to evaluate the proliferation and differentiation of keratinocytes in normal skin and scar tissue. The results showed that the structure of the skin was histologically changed in scar tissue. Collagen Ⅳ, expressed under the epidermis of normal skin, was reduced distinctly in scar tissue. Integrinβ4, expressed in the basal layer of normal skin, was found absent in the basal layer of scar tissue. Additionally, it was found that keratinocytes in scarring epidermis were more proliferative than in normal skin. These results indicate that during the skin wound healing, altered formation of BM may affect the proliferation of keratinocytes, reepithelial and tissue remodeling, and then result in scar formation. Thus, remodeling BM structure during wound repair may be beneficial for improving healing in cutaneous wounds during clinical practice.

Dimethyl sulfoxide inhibits zymosan-induced intestinal inflammation and barrier dysfunction

AIM: To investigate whether dimethyl sulfoxide(DMSO) inhibits gut inflammation and barrier dysfunction following zymosan-induced systemic inflammatoryresponse syndrome and multiple organ dysfunction syndrome.METHODS: Sprague-Dawley rats were randomly divided into four groups: sham with administration of normal saline(SS group); sham with administration of DMSO(SD group); zymosan with administration of normal saline(ZS group); and zymosan with administration of DMSO(ZD group). Each group contained three subgroups according to 4 h,8 h,and 24 h after surgery. At 4 h,8 h,and 24 h after intraperitoneal injection of zymosan(750 mg/kg),the levels of intestinal inflammatory cytokines [tumor necrosis factor-alpha(TNF-α) and interleukin(IL)-10] and oxides(myeloperoxidase,malonaldehyde,and superoxide dismutase) were examined. The levels of diamine oxidase(DAO) in plasma and intestinal mucosal blood flow(IMBF) were determined. Intestinal injury was also evaluated using an intestinal histological score and apoptosis of intestinal epithelial cells was determined by deoxynucleotidyl transferase d UTP nick end labeling(TUNEL) assay. The intestinal epithelial tight junction protein,ZO-1,was observed by immunofluorescence.RESULTS: DMSO decreased TNF-α and increased IL-10 levels in the intestine compared with the ZS group at the corresponding time points. The activity of intestinal myeloperoxidase in the ZS group was higher than that in the ZD group 24 h after zymosan administration(P < 0.05). DMSO decreased the content of malondialdehyde(MDA) and increased the activity of superoxide dehydrogenase(SOD) 24 h after zymosan administration. The IMBF was lowest at 24 h and was 49.34% and 58.26% in the ZS group and ZD group,respectively(P < 0.05). DMSO alleviated injury in intestinal villi,and the gut injury score was significantly lower than the ZS group(3.6 ± 0.2 vs 4.2 ± 0.3,P < 0.05). DMSO decreased the level of DAO in plasma compared with the ZS group(65.1 ± 4.7 U/L vs 81.1 ± 5.0 U/L,P < 0.05). DMSO significantly preserved ZO-1 protein expression and localization 24 h after zymosan administration. The TUNEL analysis indicated that the number of apoptotic intestinal cells in the ZS group was much higher than the ZD group(P < 0.05).CONCLUSION: DMSO inhibited intestinal cytokines and protected against zymosan-induced gut barrier dysfunction.

Visual function and higher order aberration after implantation of aspheric and spherical multifocal intraocular lenses:a meta-analysis

AIMTo assess the visual outcomes of aspheric multifocal intraocular lenses (IOLs) compared with spherical multifocal IOL after cataract surgery.

Astrocytes: a double-edged sword in neurodegenerative diseases

Astrocytes play multifaceted and vital roles in maintaining neurophysiological function of the central nervous system by regulating homeostasis, increasing synaptic plasticity, and sustaining neuroprotective effects. Astrocytes become activated as a result of inflammatory responses during the progression of pathological changes associated with neurodegenerative disorders. Reactive astrocytes(neurotoxic A1 and neuroprotective A2) are triggered during disease progression and pathogenesis due to neuroinflammation and ischemia. However, only a limited body of literature describes morphological and functional changes of astrocytes during the progression of neurodegenerative diseases. The present review investigated the detrimental and beneficial roles of astrocytes in neurodegenerative diseases reported in recent studies, as these cells have promising therapeutic potential and offer new approaches for treatment of neurodegenerative diseases.

Advantages of Rho-associated kinases and their inhibitor fasudil for the treatment of neurodegenerative diseases

Ras homolog(Rho)-associated kinases(ROCKs)belong to the serine-threonine kinase family,which plays a pivotal role in regulating the damage,survival,axon guidance,and regeneration of neurons.ROCKs are also involved in the biological effects of immune cells and glial cells,as well as the development of neurodegenerative disorders such as Alzheimer’s disease,Parkinson’s disease,and multiple sclerosis.Previous studies by us and others confirmed that ROCKs inhibitors attenuated the symptoms and progression of experimental models of the abovementioned neurodegenerative diseases by inhibiting neuroinflammation,regulating immune imbalance,repairing the blood-brain barrier,and promoting nerve repair and myelin regeneration.Fasudil,the first ROCKs inhibitor to be used clinically,has a good therapeutic effect on neurodegenerative diseases.Fasudil increases the activity of neural stem cells and mesenchymal stem cells,thus optimizing cell therapy.This review will systematically describe,for the first time,the effects of abnormal activation of ROCKs on T cells,B cells,microglia,astrocytes,oligodendrocytes,and pericytes in neurodegenerative diseases of the central nervous system,summarize the therapeutic potential of fasudil in several experimental models of neurodegenerative diseases,and clarify the possible cellular and molecular mechanisms of ROCKs inhibition.This review also proposes that fasudil is a novel potential treatment,especially in combination with cell-based therapy.Findings from this review add support for further investigation of ROCKs and its inhibitor fasudil for the treatment of neurodegenerative diseases.

Anterior segment dysgenesis correlation with epithelial-mesenchymal transition in Smad4 knockout mice

AIM： To explore the molecular mechanisms in lens development and the pathogenesis of Peters anomaly in Smad4 defective mice. METHODS： Le-Cre transgenic mouse line was employed to inactivate Smad4 in the surface ectoderm selectively. Pathological techniques were used to reveal the morphological changes of the anterior segment in Smad4 defective eye. Immunohistochemical staining was employed to observe the expression of E-cadherin, N- cadherin and α-SMA in anterior segment of Smad4 defective mice and control mice at embryonic （E） day 16.5. Real-time quantitative polymerase chain reaction （qPCR） was performed to detect the expression of Snail, Zebl, Zeb2 and Twist2 in lens of Smad4 defective mice and control mice at E16.5. RESULTS： Conditional deletion of Smad4 on eye surface ectoderm resulted in corneal dysplasia, iridocorneal angle closure, corneolenticular adhesions and cataract resembling Peters anomaly. Loss of Smad4 function inhibited E-cadherin expression in the lens epithelium cells and corneal epithelium cells in Smad4 defective eye. Expression of N-cadherin was upregulated in corneal epithelium and corneal stroma. Both E-cadherin and N-cadherin were down-regulated at the future trabecular meshwork region in mutant eye. The qPCR results showed that the expression of Twist2 was increased significantly in the mutant lens （P〈0.01）. CONCLUSION： Smad4 is essential to eye development and likely a candidate pathogenic gene to Peters anomaly by regulating epithelial-mesenchymal transition. Twist2 can be regulated by Smad4 and plays an essential role in lens development.

Sustained-release genistein from nanostructured lipid carrier suppresses human lens epithelial cell growth

AIM： To design and investigate the efficacy of a modified nanostructured lipid carrier loaded with genistein（Gen-NLC） to inhibit human lens epithelial cells（HLECs） proliferation.·METHODS： Gen-NLC was made by melt emulsification method. The morphology, particle size（PS）, zeta potentials（ZP）, encapsulation efficiency（EE） and in vitro release were characterized. The inhibition effect of nanostructured lipid carrier（NLC）, genistein（Gen） and Gen-NLC on HLECs proliferation was evaluated by cell counting kit-8（CCK-8） assay, gene and protein expression of the proliferation marker Ki67 were evaluated with real-time quantitative polymerase chain reaction（RT-q PCR） and immunofluorescence analyses.·RESULTS： The mean PS of Gen-NLC was 80.12±1.55 nm with a mean polydispersity index of 0.11±0.02. The mean ZP was-7.14 ±0.38 m V and the EE of Gen in the nanoparticles was 92.3% ±0.73%. Transmission electron microscopy showed that Gen-NLC displayed spherical-shaped particles covered by an outer-layer structure. In vitro release experiments demonstrated a prolonged drug release for 72 h. The CCK-8 assay results showed the NLC had no inhibitory effect on HLECs and Gen-NLC displayed a much more prominent inhibitory effect on cellular growth compared to Gen of the same concentration. The m RNA and protein expression of Ki67 in LECs decreased significantly in Gen-NLC group.·CONCLUSION： Sustained drug release by Gen-NLCs may impede HLEC growth.

Pharmacological Activity and Application of Roots, Stems and Leaves of Scutellaria baicalensis Georgi

The root of Scutellaria baicalensis Georgi is traditionally used as medicine,and it has been confirmed that S.baicalensis Georgi has flavonoid chemical constituents,pharmacological activity and cosmetic efficacy.With the extensive application of S.baicalensis Georgi roots,the resource of S.baicalensis Georgi has been increasingly short.The above-ground part of stems and leaves of S.baicalensis Georgi has also been gradually recognized and developed.Studies have found that the chemical constituents from stems and leaves of S.baicalensis Georgi are also a group of flavonoids with a lot of pharmacological activity and have a great application value.Based on this,the present review will be reported on the chemical constituents and application of the roots,stems and leaves of S.baicalensis Georgi.

Antifungal Effects of Lipopeptide Produced by <i>Bacillus amyloliquefaciens</i>BH072

An antifungal lipopeptide iturin A with strong activity against Fusarium oxysporum was produced by honey isolated strain Bacillus amyloliquefaciens BH072. For large-scale biocontrol application, the antifungal effect was deeply demonstrated by structure and mode of action. Cyclic structure and second structure were determined based on situ acid hydrolysis and Fourier Transformed-Infra Red (FT-IR) Spectra analysis. Structure of α-helix was predicted which might be associated with activity. Afterwards, antifungal mechanism of iturin A on F. oxysporum were investigated from fungal cell wall to the plasma membrane and finally to intracellular proteins by morphological, activity of alkaline phosphatase (AKP), conductivity, Malondialdehyde (MDA) and SDS-PAGE detection. Antifungal damage appears on not only the spore germination and mycelium growth of F. oxysporum, but also the leakage of cellular proteins. Moreover, growth of F. oxysporum could be inhibited in the presence of iturin A at a MIC of 2.5 mg/mL. Considered with its high production in previous work, B. amyloliquefaciens strain BH072 and iturin A might be a promising candidate for biocontrol.

Efficient and rapid conversion of human astrocytes and ALS mouse model spinal cord astrocytes into motor neuron-like cells by defined small molecules

Background: Motor neuron degeneration or loss in the spinal cord is the characteristic phenotype of motor neuron diseases or spinal cord injuries. Being proliferative and located near neurons, astrocytes are considered ideal cell sources for regenerating neurons.Methods: We selected and tested different combinations of the small molecules for inducing the conversion of human and mouse astrocytes into neurons. Microscopic imaging and immunocytochemistry analyses were used to characterize the morphology and phenotype of the induced neurons while RT-q PCR was utilized to analyze changes in gene expression. In addition, whole-cell patch-clamp recordings were measured to examine the electrophysiological properties of induced neurons.Results: The results showed that human astrocytes could be rapidly and efficiently converted into motor neuronlike cells by treatment with defined small molecules, with a yield of over 85% motor neuron-like cells attained. The induced motor neuron-like cells expressed the pan-neuronal markers TUJ1, MAP2, Neu N, and Synapsin 1 and motor neuron markers HB9, ISL1, CHAT, and VACh T. During the conversion process, the cells did not pass through a proliferative neural progenitor cell intermediate. The induced motor neurons were functional, showing the electrophysiological properties of neurons. The same chemical cocktail could induce spinal cord astrocytes from an amyotrophic lateral sclerosis mouse model carrying a SOD1 mutation to become motor neuron-like cells that exhibited a decrease in cell survival and an increase in oxidative stress compared to that observed in wild-type MNs derived from healthy mice. Moreover, the chemical induction reduced oxidative stress in the mutant astrocytes.Conclusions: The results of the present study demonstrated the feasibility of chemically converting human and mouse astrocytes into motor neuron-like cells that are useful for neurodegenerative disease modeling and regenerative medicine.

Acute lens opacity induced by different kinds of anesthetic drugs in mice

AIM: To study whether specific anesthetic drugs or tear layer evaporation was primarily responsible for the acute cataract and what the change of lens structure is in anesthetized mice.METHODS: Five groups were set up in the experiment: Group A(topicamide and phenylephrine mixed eye drop+ chloral hydrate), Group B(tropicamide and phenylephrine mixed eye drop+sevoflurane), Group C(tropicamide and phenylephrine mixed eye drop), Group D(topicamide and phenylephrine mixed eye drop+chloral hydrate, carbomer eye drop in the right eyes), and Group E(tropicamide and phenylephrine mixed eye drop+sevoflurane, carbomer eye drop in the right eyes). A simple classification system was used to assess the severity of lens opacity. And a numerical value from 0 to 3 to each grade was assigned for the cataract index calculation and data analysis. The gross appearance and time course of development of lens opacity were assessed. Hematoxylin and eosin staining was used to observe the lens structure changes in the reversible cataract.RESULTS: Tropicamide did not induce lens opacification in mice. Lens opacity caused by inhaled sevoflurane was similar to injected cholral hydrate. Both inhaled-anestheticinduced lens opacity and injected-anesthetic-induced lens opacity could be prevented by carbomer eye drop. In the severe opacity lens, a wide range of lens fiber cell structure had disordered. The fiber cells became uneven thickness.CONCLUSION: The acute reversible lens opacity can unilaterally develop or be induced by a local cause. The structure of lens fiber cells changed in the lens opacity which may influence the permanent connection of the lens fiber cells. This study was not only of practical significance to help maintain lens transparency for eye research, but also of the deeper consideration about the reversible lens opacification phenomenon.

Role of Smad4 from ocular surface ectoderm in retinal vasculature development

●AIM:To investigate how signals from lens regulate retinal vascular development and neovascularization.●METHODS:Le-Cre transgenic mouse line was employed to inactivate Smad4 in the surface ectoderm selectively.Standard histological and whole-mount retina staining were employed to reveal morphological changes of retinal vasculature in Smad4 defective eye.cDNA microarray and subsequent analyses were conducted to investigate the molecular mechanism underlying the vascular phenotype.Quantitative polymerase chain reaction(qPCR)was carried out to verify the microarrays results.●RESULTS:We found that inactivation of Smad4 specifically on surface ectoderm leads to a variety of retinal vasculature anomalies.Microarray analyses and qPCR revealed that Sema3 c,Sema3 e,Nrp1,Tie1,Sox7,Sox17,and Sox18 are significantly affected in the knockout retinas at different developmental stages,suggesting that ocular surface ectoderm-derived Smad4 can signal to the retina and regulates various angiogenic signaling in the retina.●CONCLUSION:Our data suggest that the cross-talk between ocular surface ectoderm and retina is important for retinal vasculature development,and Smad4 regulates various signaling associated with sprouting angiogenesis,vascular remodeling and maturation in the retina of mice.