Dedifferentiation-like reprogramming of degenerative nucleus pulposus cells into notochordal-like cells by defined factors.

The extensive degeneration of functional somatic cells and the depletion of endogenous stem/progenitor populations present significant challenges to tissue regeneration in degenerative diseases. Currently, a cellular reprogramming approach enabling directly generating corresponding progenitor populations from degenerative somatic cells remains elusive. The present study focused on intervertebral disc degeneration (IVDD) and identified a three-factor combination (OCT4, FOXA2, TBXT (OFT)) that could induce the dedifferentiation-like reprogramming of degenerative nucleus pulposus cells (dNPCs) toward induced notochordal-like cells (iNCs). Single-cell transcriptomics dissected the transitions of cell identity during reprogramming. Further, OCT4 was found to directly interact with bromodomain PHD-finger transcription factor (BPTF) to remodel the chromatin during the early phases, which was crucial for initiating this dedifferentiation-like reprogramming. In rat models, intradiscal injection of adeno-associated virus carrying OFT generated iNCs from in situ dNPCs and reversed IVDD. These results collectively present a proof-of-concept for dedifferentiation-like reprogramming of degenerated somatic cells into corresponding progenitors through the development of a factor-based strategy, providing a promising approach for regeneration in degenerative disc diseases.

Methylation in cornea and corneal diseases: a systematic review.

Corneal diseases are among the primary causes of blindness and vision loss worldwide. However, the pathogenesis of corneal diseases remains elusive, and diagnostic and therapeutic tools are limited. Thus, identifying new targets for the diagnosis and treatment of corneal diseases has gained great interest. Methylation, a type of epigenetic modification, modulates various cellular processes at both nucleic acid and protein levels. Growing evidence shows that methylation is a key regulator in the pathogenesis of corneal diseases, including inflammation, fibrosis, and neovascularization, making it an attractive potential therapeutic target. In this review, we discuss the major alterations of methylation and demethylation at the DNA, RNA, and protein levels in corneal diseases and how these dynamics contribute to the pathogenesis of corneal diseases. Also, we provide insights into identifying potential biomarkers of methylation that may improve the diagnosis and treatment of corneal diseases.

CircRNA expression profiles and regulatory networks in the vitreous humor of people with high myopia.

Myopia is a global health and economic issue. Circular RNAs (circRNAs) have been shown to play an important role in the pathogenesis of many ocular diseases. We first evaluated the circRNA profiles and possible roles in vitreous humor samples of individuals with high myopia by a competitive endogenous RNA (ceRNA) array. Vitreous humor samples were collected from 15 high myopic (5 for ceRNA array, and 10 for qPCR) and 15 control eyes (5 for ceRNA array, and 10 for qPCR) with idiopathic epiretinal membrane (ERM) and macular hole (MH). 486 circRNAs (339 upregulated and 147 downregulated) and 264 mRNAs (202 upregulated and 62 downregulated) were differentially expressed between the high myopia and control groups. The expression of hsa_circ_0033079 (hsa-circDicer1), hsa_circ_0029989 (hsa-circNbea), hsa_circ_0019072 (hsa-circPank1) and hsa_circ_0089716 (hsa-circEhmt1) were validated by qPCR. Pearson analysis and multivariate regression analysis showed positive and significant correlations for axial length with hsa-circNbea and hsa-circPank1. KEGG analysis showed that the target genes of circRNAs were enriched in the mTOR, insulin, cAMP, and VEGF signaling pathways. GO analysis indicated that circRNAs mainly targeted transcription, cytoplasm, and protein binding. CircRNA-associated ceRNA network analysis and PPI network analysis identified several critical genes for myopia. The expression of circNbea, circPank1, miR-145-5p, miR-204-5p, Nras, Itpr1 were validated by qPCR in the sclera of form-deprivation myopia (FDM) mice model. CircPank1/miR-145-5p/NRAS and circNbea/miR-204-5p/ITPR1 were identified and may be important in the progression of myopia. Our findings suggest that circRNAs may contribute to the pathogenesis of myopia and may serve as potential biomarkers.

Agreement Between Single Raters and Team Rating When Applying the International Classification of Functioning, Disability and Health's Rehabilitation Set.

OBJECTIVE: To quantify the agreement between functional assessments by a single rater and a team using the Chinese version of the International Classification of Functioning, Disability and Health Rehabilitation Set in a clinical situation. DESIGN: Inter-rater, multi-centre agreement study. SUBJECTS: A total of 193 adult inpatients admitted to 5 rehabilitation centres at 5 hospitals in China Methods: The Chinese version of the International Classification of Functioning, Disability and Health Rehabilitation Set was used by either a single rater or a team to assess 193 patients at 5 Chinese hospitals. Percentage of agreement and quadratic-weighted kappa coefficients were computed. Evaluation times were compared with paired t-tests. RESULTS: The mean team and individual evaluation times were not significantly different. The percentage of agreement ranged from 46.1% to 94.2% depending on the item, and the quadratic-weighted kappas ranged from 0.43 to 0.92. Eight categories (26.6%) showed a weighted kappa exceeding 0.4, 11 others (36.7%) exceeded 0.6, and another 11 (36.7%) produced kappas of more than 0.8. CONCLUSION: Either a single rater or a team of raters can produce valid and consistent ratings when using the Chinese version of the International Classification of Functioning, Disability and Health Rehabilitation Set to assess patients in a rehabilitation department. The team rating approach is suitable for clinical application.

Adverse childhood experiences and depressive symptoms among lesbian and bisexual women in China.

BACKGROUND: Despite the relationship between Adverse childhood experiences (ACEs) and depressive symptoms, which has been well researched in general populations, little is known about homosexual and bisexual populations, especially lesbian and bisexual women in China. This study aims to investigate the prevalence of ACEs and depressive symptoms and to analyze the relationship between them among lesbian and bisexual women in China. METHODS: The eligible participants were aged 16 years or older who report their sexual orientation as homosexual or bisexual. The data was collected through anonymous questionnaires with the help of Lespark in Beijing from July 18 to December 29, 2018, and all participants had informed consent to this study. Univariate analysis and multiple linear regression analyses were performed to explore the relationship between ACEs and depressive symptoms among lesbian and bisexual women. All statistical analyses were conducted by the software of SPSS 22.0. RESULTS: Among 301 lesbian and bisexual women, 81.4% were lesbian, 18.4% were bisexual women, and the majority were 21-30 years. As for ACEs, 51.5% reported at least one ACE, in which emotional neglect (22.6%) and emotional abuse (22.3%) were common ACEs. As for depressive symptoms of lesbian and bisexual women, the detection rate was 56.1%. The multiple linear regression analyses showed that abuse (ß = 2.95, 95%CI:1.07-4.83) and neglect (ß = 3.21, 95%CI:1.09-5.31) were positively associated with depressive symptoms and lesbian and bisexual women with three (ß = 4.11, 95%CI: 0.99-7.22) or more (ß = 6.02, 95%CI: 3.23-8.78) ACEs suffered from more depressive symptoms than others. CONCLUSION: Adverse childhood experiences (ACEs) and depressive symptoms were at high prevalence among lesbian and bisexual women in China. ACEs were associated with depressive symptoms, especially childhood abuse and neglect experiences that have a significant effect on lesbian and bisexual women mental health.

Blue light impairs cornea and corneal wound healing by downregulating VCAM1 partly.

This study aimed to investigate the effects of long-term pollution from different wavelengths of light on the corneal epithelium (CE) and identify potential biomarkers. Rabbits were exposed to red, green, blue, white, and environmental light for 6 weeks. The CE was assessed using various techniques such as fluorescein sodium staining, transcriptome sequencing, electron microscopy, and molecular assays. In human corneal epithelial cells (hCECs), the downregulation of vascular cell adhesion molecule 1 (VCAM1) in response to blue light (BL) pollution was observed. This downregulation of VCAM1 inhibited migration, increased reactive oxygen species (ROS) levels, and apoptosis, and inhibited the AKT/p70 S6 kinase cascade in hCECs. Animal experiments confirmed that BL pollution caused similar effects on the rabbit cornea, including increased ROS production, apoptosis, delayed wound healing, and decreased VCAM1 expression. Overall, BL-induced VCAM1 downregulation may impair CE and wound healing and promote ROS and apoptosis in vitro and in vivo.

Long-term blue light exposure impairs mitochondrial dynamics in the retina in light-induced retinal degeneration in vivo and in vitro.

Long-term light exposure, especially in the spectrum of blue light, frequently causes excessive oxidative stress in dry age-related macular degeneration (AMD). Here, to gain insight into the underlying mechanism, we focused on mitochondrial dynamics alterations under long-term exposure to blue light in mouse and retinal cells. Six-month-old C57BL/6 mice were exposed to blue light (450 nm, 800 lx) for 2 weeks. The phenotypic changes in the retina were assayed using haematoxylin-eosin staining and transmission electron microscopy. Long-term blue light exposure significantly thinned each retinal layer in mice, induced retinal apoptosis and impaired retinal mitochondria. A retinal pigment epithelial cell line (ARPE-19) was used to verify the phototoxicity of blue light. Flow cytometry, immunofluorescence and MitoSox Red probe experiments confirmed that more total and mitochondria-specific ROS were generated in the blue light group than in the control group. Mito-Tracker Green probe showed fragmented mitochondrial morphology. The western blotting results indicated a significant increase in DRP1, OMA1, and BAX and a decrease in OPA1 and Bcl-2. In conclusion, long-term exposure to blue light damaged the retinas of mice, especially the ONL and RPE cells. There was destruction and dysfunction of mitochondria in RPE cells in vivo and in vitro. Mitochondrial dynamics were disrupted with characteristics of fusion-related obstruction after blue-light irradiation.

Blue light attenuates TGF-ß2-induced epithelial-mesenchymal transition in human lens epithelial cells via autophagy impairment.

BACKGROUND: Pathogenesis of posterior capsular opacification (PCO) was related to pathological epithelial-mesenchymal transition (EMT) of lens epithelial cells (LECs). It has been reported that blue light could have an effect on EMT. This study aims to elucidate the role and potential mechanism of autophagy in EMT after blue light exposure in LECs. METHODS: HLE-B3 cells were treated with TGF-ß2 with different concentration and time to induce EMT as a model of PCO in vitro. Cells were exposed to blue light with or without TGF-ß2. The expression levels of EMT-associated markers were analyzed by qRT-PCR, western blotting and cell migration ability was determined by transwell migration assay and wound healing assay. The expressions of autophagy-related proteins were analyzed by western blotting, immunofluorescence and transmission electron microscopy. Rapamycin and chloroquine were utilized in cells for autophagy activation and inhibition. RESULTS: TGF-ß2 induced autophagy activation during EMT progression in HLE-B3 cells in a dose- and time-dependent manner. Blue light exposure inhibited TGF-ß2-induced EMT characterized by inhibited expression of EMT related markers and reduced migration capacity. Meanwhile, blue light exposure impaired autophagy activated by TGF-ß2. Furthermore, Autophagy activation with rapamycin rescued EMT attenuated by blue light. Autophagy inhibition with chloroquine reduced TGF-ß2-induced EMT in HLE-B3 cells. CONCLUSION: Blue light exposure had inhibited effects on TGF-ß2-induced EMT in LECs through autophagy impairment, which provides a new insight on prevention and treatment of PCO.

Comparative physiological and soil microbial community structural analysis revealed that selenium alleviates cadmium stress in Perilla frutescens.

Cadmium (Cd) toxicity not only affects plant growth and development, but also affects human health through the food chain. Several studies have demonstrated that Selenium (Se) alleviates Cd stress in plants; however, whether and how Se-alleviated Cd stress by regulating the structure of soil microbial community remain largely unclear. Here, we investigated the alleviating effects of exogenous applied Se (foliar spraying or root application) on plant growth under Cd stress in perilla (Perilla frutescens L.) by measuring the biomass, photosynthetic fluorescence parameters, root cell wall components and soil microbial community structure and diversity. Under Cd stress, perilla seedlings supplemented with Se increased chlorophyll content. Foliar spraying Se increased the levels of relative chlorophyll content (ΦII), photosynthetic system II (ΦPSII) and electron transport rate (ETR) in perilla leaves under Cd stress; while, root application of Se increased the levels of photosynthetic rate (Pn), stomatal conductance (Gs), transpiration rate (Tr), water use efficiency (WUE) and stomatal limitation value (Ls) under Cd stress. Compared with Cd toxicity alone, root application of Se increased the contents of hemicellulosic 1 and hemicellulosic 2 in the cell wall of perilla roots. Cd toxicity or root application of Se did not affect soil bacterial community diversity. Root application of Se increased the relative abundance of Proteobacteria, Bacteroidetes, Fibrobacteres, Sphingomonas and Nitrosospira in Cd-contaminated soil, and thereby improving soil microbial community structure, finally promoting the growth of perilla seedlings.

A Review of Intraocular Pressure (IOP) and Axial Myopia.

The pathogenesis of myopia is driven by genetic and environmental risk factors. Accommodation not only alters the curvature and shape of the lens but also involves contraction of the ciliary and extraocular muscles, which influences intraocular pressure (IOP). Scleral matrix remodeling has been shown to contribute to the biomechanical susceptibility of the sclera to accommodation-induced IOP fluctuations, resulting in reduced scleral thickness, axial length (AL) elongation, and axial myopia. The rise in IOP can increase the burden of scleral stretching and cause axial lengthening. Although the accommodation and IOP hypotheses were proposed long ago, they have not been validated. This review provides a brief and updated overview on studies investigating the potential role of accommodation and IOP in myopia progression.

Quantitative proteomic analysis of scleras in guinea pig exposed to wavelength defocus.

Myopia is the most common optical disorder in the world, and wavelength defocus induced ametropia and myopia have attracted great attention. The objective was to identify and quantify scleral proteins involved in the response to the wavelength defocus. Guinea pigs were randomly divided into 3 groups that received different lighting conditions for 8 weeks: white light, short wavelength light, and long wavelength light. Refraction and axial length were measured, Hematoxylin-Eosin staining and transmission electron microscope were adopted to observe the scleral structure, and scleral proteome was also detected to analyze protein abundance by employing TMT labeling method. After light stimulation, the long- and short -wavelength light induced myopic and hyperopic effect on the guinea pig's eye and induced distinct protein signature, respectively. 186 dyregulated proteins between the short- and long-wavelength group were identified, which were mainly located in extracellular region and involved in metabolic process. We also found that 5 proteins in the guinea pigs scleras in response to wavelength defocus were also human myopic candidate targets, suggesting functional overlap between dyregulated proteins in scleral upon exposure to wavelength defocus and genes causing myopia in humans. SIGNIFICANCE: Wavelength defocus induces refractive errors and leads to myopia or hyperopia. However, sclera proteomics respond to wavelength defocus is lacking, which is crucial to understanding how wavelength defocus influences refractive development and induces myopia. In this proteome analysis, we identified unique protein signatures response to wavelength defocus in sclera of guinea pigs, identified potential mechanisms contributing to myopia formation, and found that several human myopia-related genes may involve in response to wavelength defocus. The results of this study provide a foundation to understand the mechanisms of myopia and wavelength defocus induced ametropia.

Psychosocial characteristics and HIV-related sexual behaviors among cisgender, transgender, and gender non-conforming MSM in China.

BACKGROUND: While a growing number of studies focus on men who have sex with men (MSM), they typically ignore the heterogeneity of gender minorities within the MSM population. The recognition of new sub-groups among gender minorities (i.e., transgender and gender non-conforming), who also identify as MSM, play a considerable role in new HIV infections in China. Information on the psychosocial factors and HIV-related sexual behaviors require further consideration to understand the prevalence of HIV infection among MSM within these gender minority sub-groups. METHODS: From September 2017 to January 2018, MSM without HIV were recruited in Wuhan, Nanchang, and Changsha cities in China. Participants were asked to fill out a structured self-administered questionnaire to assess depression, perceived social support, resilience, identity concealment, and HIV-related risky sexual behaviors. RESULTS: A total of 715 MSM completed the structured questionnaire, the number of MSM identifying as gender minorities were 63 and accounted for 8.8% of the population. Compared to the cisgender MSM population, transgender MSM were more likely to have a one-night stand/occasional partner (AOR = 3.49, 95% CI =1.02-11.98), to have sex after drug use in the past 6 months (AOR = 2.57, 95%CI =1.05-6.29), and to have reported a significantly lower likelihood of identity concealment (mean difference = - 3.30, 95%CI = -5.86, - 0.74, P = 0.01). CONCLUSIONS: The findings highlight the significance of providing targeted interventions for different gender minorities within the MSM population. Research is required to further understand the relationship between gender identity, mental health, and HIV-related sexual behaviors.

A team approach to applying the International Classification of Functioning, Disability and Health Rehabilitation set in clinical evaluation.

OBJECTIVE: To develop a team approach to applying the International Classification of Functioning, Disability and Health Rehabilitation Set (ICF-RS) in clinical evaluation. DESIGN: A Delphi study. SUBJECTS: Experts from rehabilitation institutions in China including physicians, nurses, physiotherapists and occupational therapists. METHODS: A 2-round Delphi survey and expert panel discussion were used to generate the team approach. Firstly, the candidate types of professionals for team rating were chosen through expert panel discussion. A carefully selected group of participants was then asked to score the suitability of physicians, nurses, or other candidate therapists for each category's rat-ing, applying the International Classification of Functioning, Disability and Health Rehabilitation Set in clinical evaluation. After initial assignment of cate-gory to types of professionals, a second round Delphi survey was conducted to quantify the professionals' agreement with the category assignments and generate a final team evaluation approach. RESULTS: Thirty of the category assignments achiev-ed consensus. The final team evaluation approach assigned 6 categories to physicians to evaluate, 7 categories to nurses, 9 categories to physiotherapists, and 8 to occupational therapists. CONCLUSION: Such a team evaluation approach could facilitate implementation of the ICF-RS in clinical settings and provide a more convenient assessment tool for professionals.

n-butanol extract from Folium isatidis inhibits the lipopolysaccharide-induced downregulation of CXCR1 and CXCR2 on human neutrophils.

Neutrophils, immune cells crucial for protecting against invading pathogens, are important in sepsis. Neutrophil migration is regulated by chemokine receptors and their cognate ligands. Our previous study investigated the effect of n­butanol extract from Folium isatidis on lipopolysaccharide (LPS)­induced septic shock. The present study stimulated neutrophils with LPS to explore the influence of LPS on cell. Neutrophils were then pretreated with n­butanol extract from Folium isatidis followed by LPS to examine the effect of this extract on neutrophil chemotaxis. The results showed that LPS decreased the expression levels of CXC­chemokine receptor (CXCR)1, CXCR2 and L­selectin (CD62L), and increased the expression of interleukin­8 (IL­8) by neutrophils. The addition of n­butanol extract from Folium isatidis inhibited this LPS­induced downregulation of CXCR1, CXCR2 and CD62L, and decreased the expression of IL­8 on neutrophils. In addition, n­butanol extract promoted myeloperoxidase activity in neutrophils. Taken together, LPS downregulated the expression of chemokine receptors, leading to the failure of neutrophils to migrate to sites of infection. The addition of n­butanol extract, which promoted the ability of neutrophils to migrate, is a natural product and potential therapeutic agent with which to target neutrophil chemotaxis during LPS stimulation.

Molecular genetic analysis and phenotypic characteristics of a consanguineous family with glycogen storage disease type Ia.

Glycogen storage disease type­Ia (GSD­Ia) is a rare autosomal recessive disease caused by a mutation in the gene encoding glucose­6­phosphate­α (G6PC). The present study reported the case of a 3­month­old female Chinese patient with GSD­Ia born to consanguineous parents. The aim of the present study was to identify the precise mutation of the G6PC gene associated with this family and to describe the phenotypic characteristics of the patient. A comprehensive examination was performed on the patient, including physical examination, vein blood gas analysis, abdominal sonography and biochemical analyses. In addition, gene sequencing was performed on the coding region of the G6PC gene to identify the mutation. The patient was diagnosed with GSD­Ia and a G6PC missense mutation of c.518T>C (p.L173P) located in a highly conserved area was identified. The mutation is in a non­helical region of the protein, which previous studies have suggested should result in a lesser effect on G6PC enzymatic activity and milder phenotypic characteristics compared with mutations located in helical regions. However, the severity of the disease phenotype in the subject of the present study was inconsistent with that predicted from her genotype. The patient suffered from serious hypoglycemia, lactic acidosis, increased triglycerides, hepatic dysfunction, clear hepatomegaly and nephromegaly. The incidence of the p.L173P mutation may be relatively high in the Chinese population. Knowledge of the various phenotypic presentations of the p.L173P mutation may beneficial for future investigations.

MicroRNA-221 accelerates the proliferation of laryngeal cancer cell line Hep-2 by suppressing Apaf-1.

Laryngeal cancer is one of the most commonly occurring malignant cancers of the head and neck region. In the present study, we investigated the roles of miR-221 in laryngeal squamous cell carcinoma cell line, Hep-2. We examined the function and mechanism of miR-221 in Hep-2 cells using techniques of cell biology and molecular pathology, such as western blotting, quantitative PCR, immunohistochemical staining and flow cytometry. Using a luciferase assay, the apoptotic protease activating factor-1 (Apaf-1) mRNA 3'-UTR was shown to have complementary binding sites using bioinformatics prediction software including TargetScan, PicTar and miRanda. In conclusion, our results showed that miR-221 inhibition caused elevated expression levels of the Apaf-1 apoptotic pathway proteins caspase-3, -8 and -9. miR-221 may therefore be used as a novel therapeutic target for laryngeal cancer.