RESUMO
Akt is a central node of many signaling pathways, which plays important roles in cell survival, proliferation, migration, metabolism and collagen synthesis. Conjunctivochalasis (CCH) is one of the most common age-related ocular superficial diseases related to abnormalities in conjunctival extracellular matrix. Here, we studied the role of Akt regulating collagens and MMPs in the pathogenesis of CCH. Primary conjunctival fibroblasts were obtained from CCH patients (n = 13) and age-matched normal controls (n = 10). The levels of Akt, collagen type I, collagen type III, MMP1, and MMP3 were determined by Western blot, qRT-PCR, immunohistochemistry, and immunofluorescence staining. Normal control conjunctival fibroblasts were treated with Akt inhibitor A6730, and CCH fibroblasts were transfected with Akt overexpression vector. The expression of Akt in CCH was significantly lower than that in normal control of conjunctival tissues and cultured fibroblasts. Blocking Akt signaling with Akt inhibitor could inhibit the expression of collagen type I and collagen type III and upregulate the expression of MMP1 and MMP3. Meanwhile, compared with CCH fibroblasts transfected with control mimics, the protein and mRNA expression of collagen type I and collagen type III were increased significantly in Akt overexpression group, while the results of MMP1 and MMP3 in transfected fibroblasts were opposite. Taken together, Akt upregulated the expression of collagen type I and collagen type III and downregulated the expression of MMP1 and MMP3. Akt signaling pathway could provide a direct negative contribution to CCH and might be an attractive target for CCH therapy.
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Colágeno , Doenças da Túnica Conjuntiva , Metaloproteinase 1 da Matriz , Metaloproteinase 3 da Matriz , Proteínas Proto-Oncogênicas c-akt , Humanos , Células Cultivadas , Colágeno/metabolismo , Colágeno Tipo I/genética , Colágeno Tipo I/metabolismo , Colágeno Tipo III , Doenças da Túnica Conjuntiva/metabolismo , Fibroblastos/metabolismo , Metaloproteinase 1 da Matriz/genética , Metaloproteinase 1 da Matriz/metabolismo , Metaloproteinase 3 da Matriz/genética , Metaloproteinase 3 da Matriz/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismoRESUMO
Conjunctivochalasis (CCH) is a common ocular disease, especially in aged people. However, the molecular mechanism of CCH on transcriptional level has been unclear. In this study, we characterized the transcriptional landscape of human conjunctiva and compared the transcriptome between normal persons (nâ¯=â¯10) and CCH patients (nâ¯=â¯11). Illumina RNA sequencing (RNA-seq) was performed to obtain transcriptional data, and these data were analyzed using various bioinformatics methods, including read mapping, the analysis of gene expression, gene ontology (GO) analysis and Kyoto Encyclopedia of Gene and Genomes (KEGG) metabolic pathway analysis. Additionally, expression patterns of 20 dysregupated genes were validated by qRT-PCR. RNA-seq result showed that clean ratios of 21 samples were more than 95% and more than 92% of all clean reads (32-41 million reads) were mapped to human genome sequence. There were 175 up-regulated genes and 582 down-regulated genes identified in CCH compared to normal persons. Among down-regulated genes in CCH, many genes were related with cell cycle and proliferation, such as BUB1, CCNB1, CCNB2 and CENPA, which might disturb cell growth and proliferation. In addition, several down-regulated genes were associated with keratinization and differentiation of epidermal cells, such as SPRR1A, SPRR1B, and CALML5. In over-expressed genes, CALML6 might play important roles on the development of CCH. The results of qRT-PCR confirmed the accuracy and credibility of RNA-Seq analysis. This study provided a lot of valuable information about pathogenic mechanism of CCH, which could be used to better study CCH in the future.
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Doenças da Túnica Conjuntiva/genética , Proteínas do Olho/genética , Perfilação da Expressão Gênica , Análise de Sequência de RNA , Transcriptoma/genética , Idoso , Idoso de 80 Anos ou mais , Túnica Conjuntiva/metabolismo , Feminino , Voluntários Saudáveis , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Masculino , Pessoa de Meia-Idade , Análise de Componente Principal , Reação em Cadeia da Polimerase em Tempo RealRESUMO
To investigate the distribution of traditional Chinese medicine (TCM) Syndromes of diabetic retinopathy and explore the correlation between various symptoms. Based on the literature in databases of China Journal Full-text Database (CNKI), Wanfang database, VIP network, China biomedical literature database (CBM) and PubMed, SPSS 20.0 and IBM SPSS Modeler 14.1 software were used to analyze the location of disease, symptoms, tongue and pulse, and syndrome type distribution through frequency statistics. In addition, association rule algorithm was used to explore the basic rules for underlying symptoms combinations of diabetic retinopathy. A total of 560 articles were retrieved, and a total of 240 articles met the inclusion and exclusion criteria. In these documents, 62 types of TCM syndromes were found, involving 9 820 cases, 201 symptoms, 29 tongue conditions, and 36 pulses conditions; the first two locations for the disease were liver and kidney. Then the association rule analysis of high frequency symptoms was used to dig out 15 groups of latent syndrome, and 3 underlying symptom combinations among high frequency symptoms, tongue conditions and pulse conditions. The results of the study showed that Qi and yin deficiency was most common for the diabetic retinopathy, and the location of the disease was closely related to liver and kidney. In addition, these high-frequency symptoms and tongue conditions, pulse conditions, and underlying symptom combinations can occur as main symptoms at diagnosis, providing reference for us to study the epidemiology of PRO scale of the disease. They can also increase the weight of these symptoms directly as the main symptoms, which can be also used as an alternative entry pool for TCM syndrome diagnostic scale, laying foundation for the construction and optimization of TCM symptom database of diabetic retinopathy.
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Retinopatia Diabética/diagnóstico , Medicina Tradicional Chinesa , China , Diagnóstico Diferencial , Humanos , Qi , Síndrome , Deficiência da Energia YinRESUMO
PURPOSE: The aim of this study was to observe the expression of interleukin (IL)-17 and intercellular adhesion molecule (ICAM)-1 in conjunctivochalasis (CCH) and to analyze the correlations between cytokines and the severity of CCH. METHODS: Serum samples were collected from 22 patients with CCH and 18 normal controls (NCs). The Ocular Surface Disease Index, tear film break-up time, Schirmer I test, and corneal fluorescein staining were used to evaluate the ocular surface signs and symptoms. The concentrations of IL-17, IL-23, and ICAM-1 in serum and cellular supernatants were measured by enzyme-linked immunosorbent assays, and the gene expression levels of cytokines were measured by a quantitative real-time polymerase chain reaction. The relationships between serum concentrations of IL-17, IL-23, and ICAM-1 with clinical ocular surface parameters in CCH were analyzed using the Spearman correlation analysis. RESULTS: The concentrations of IL-17 and ICAM-1 in serum and cellular supernatants of CCH were significantly higher than those of NCs (all P < 0.001). The concentrations of IL-23 in serum and cellular supernatants of CCH showed no significant difference from those of NCs ( P > 0.05). The mRNA expression levels of IL-17 and ICAM-1 in conjunctival fibroblasts of CCH were significantly higher than those of NCs (all P < 0.001). The mRNA expression of IL-23 in conjunctival fibroblasts of CCH was higher than that of NCs, without a significant difference ( P > 0.05). Furthermore, the serum concentrations of IL-17 and ICAM-1 were positively correlated with Ocular Surface Disease Index and fluorescein staining (all P < 0.05), and negatively correlated with break-up time and Schirmer I test of CCH (all P < 0.05). CONCLUSIONS: The expression levels of IL-17 and ICAM-1 were significantly increased in CCH serum and associated with the disease severity. We postulate that IL-17 and ICAM-1 may play a role in the pathogenesis of CCH. IL-17 and ICAM-1 antagonists may be a potential treatment option for CCH in the future.
Assuntos
Doenças da Túnica Conjuntiva , Molécula 1 de Adesão Intercelular , Humanos , Molécula 1 de Adesão Intercelular/genética , Interleucina-17 , Doenças da Túnica Conjuntiva/patologia , Citocinas , Fluoresceína , Interleucina-23 , RNA Mensageiro/genéticaRESUMO
OBJECTIVE: To investigate the relationship between the occurrence and development of conjunctivochalasis and bulbar conjunctival lymphangiectasia. METHODS: Case control study. One hundred cases with conjunctivochalasis treated from January to March 2012 were selected to study, and 100 cases with no conjunctivochalasis as the control group at the same time. To observe bulbar conjunctiva lymphatic duct dilatation using slit lamp microscope, analysis bulbar conjunctiva and fascia images by OCT scanning, and ablate lymphatic of conjunctival tissue for pathologic examine. RESULTS: Twenty-nine eyes of the bulbar conjunctiva lymphangiectasia associated with 100 cases (183 eyes) conjunctivochalasis patients, accounting for 15.84%; 8 eyes of the ball conjunctival lymphatic dilation in control group of 100 cases ( 200 eyes), accounting for 4.00%. The difference between the two groups was statistically significant (χ(2) = 15.36, P < 0.001). OCT scanning showed that lymphangiectasia of the conjunctiva is at the subcutaneous mainly, some in the conjunctival lamina propria. They are border-clear, full-filled fluid, single-lumen or multi lumens, not involving the fascia. The histopathological examination showed that the lamina propria of the bulbar conjunctiva mildly chronic inflammatory changes accompanied by a large number of lymphangiectasia. CONCLUSION: Bulbar conjunctival lymphangiectasia may be one of the reasons for the conjunctivochalasis.
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Doenças da Túnica Conjuntiva/patologia , Linfangiectasia/patologia , Idoso , Idoso de 80 Anos ou mais , Estudos de Casos e Controles , Feminino , Humanos , Masculino , Pessoa de Meia-IdadeRESUMO
BACKGROUND: Qijing Mingmu decoction (QJMM), a compound Chinese medicine preparation, which consists of Lycium barbarum, Polygonatum, Ophiopogon japonicus, Poria cocos, Glycyrrhiza, Eclipta prostrata and Ligusticum striatum, has been confirmed to be effective for the treatment of conjunctivochalasis (CCH) in clinic and reduce cellular senescence. However, the underlying mechanism is still unknown. Our previous study revealed that p38-mediated cellular senescence contributed to the pathogenesis of CCH. METHODS: To explore whether p38 might be the potential therapeutic target of QJMM for CCH, CCH fibroblasts were treated with QJMM granule and then the effect of QJMM granule on the expression and promoter activity of p38α was determined by western blot and dual luciferase reporter gene assay, respectively. Meanwhile, the influence of QJMM granule on cell proliferation, oxidative stress, cellular senescence and the expression of the cellular senescence-associated genes were measured by corresponding methods. RESULTS: QJMM granule significantly decreased the protein expression of p38α and p-p38α in CCH fibroblasts in a dose-dependent manner and inhibited p38α promoter activity. QJMM granule as well as the p38 inhibitor SB203580 reduced the level of reactive oxygen species and increased the activity of superoxide dismutase in CCH fibroblasts. QJMM granule and SB203580 promoted cell proliferation and reduced the percentage of SA-ß-Gal-positive cells. The mRNA and protein expression of p53 and p21 was remarkably down-regulated by QJMM granule as well as SB203580 and that of SMP30 was up-regulated in CCH fibroblasts. CONCLUSIONS: Our findings demonstrated that QJMM granule was effective for alleviating cellular senescence of CCH fibroblasts by p38 MAPK signaling and the followed p53/p21 signaling.
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Bioensaio , Proteína Supressora de Tumor p53 , Western Blotting , Proliferação de Células , Senescência CelularRESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: Qi Jing Mingmu (QJMM) decoction is a traditional Chinese medicine that has been widely used for the clinical treatment of conjunctivochalasis (CCH). It is an effective treatment to relieve ocular symptoms including improving tear film and promoting tear secretion. However, its effects and molecular mechanisms need to be elucidated. AIM OF THE STUDY: To determine whether QJMM decoction affected T helper 17 (Th17) cell differentiation of CCH patients. MATERIALS AND METHODS: Blood samples and conjunctival tissues were collected from CCH patients and normal controls. The fibroblasts were separately induced, and CD4+ T cells were incubated with increasing concentrations of QJMM decoction and co-cultured with CCH fibroblasts. Th17 cell numbers were then analyzed using flow cytometry. Serum levels of interleukin 17 (IL-17) and IL-22 were detected using enzyme-linked immunosorbent assays. The expressions of signal proteins and genes were detected using western blotting and quantitative real-time PCR. RESULTS: Compared with normal controls, Th17 cell numbers and serum levels of IL-17 and IL-22 were elevated in patients with CCH. QJMM decoction down-regulated the expressions of IL-17, IL-22, and STAT3 of CD4+T cells from CCH patients, suggesting that QJMM decoction impeded Th17 cell differentiation. QJMM decoction-treated CD4+ T cells inhibited the expression of p38 in CCH fibroblasts. CONCLUSION: QJMM decoction inhibited Th17 cell differentiation of CD4+T cells from CCH patients, and QJMM decoction-treated CD4+T cells down-regulated the p38 signal pathway in CCH fibroblasts. Our study showed that Th17 cells may be good candidates for clinical treatment of CCH.
Assuntos
Doenças da Túnica Conjuntiva , Interleucina-17 , Humanos , Diferenciação Celular , Doenças da Túnica Conjuntiva/metabolismo , Regulação para Baixo , Fibroblastos , Interleucina-17/metabolismo , Qi , Transdução de Sinais , Células Th17 , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismoRESUMO
Purpose: To investigate the impacts of sensory impairments (SIs) including single vision impairment (SVI), single hearing impairment (SHI) and dual sensory impairment (DSI) on subjective wellbeing measurements including life expectancy (LE), life satisfaction (LS) and self-rated health (SRH) in middle-aged and older Chinese population. Methods: We obtained data from the China Health and Retirement Longitudinal Survey (CHARLS). In total, 9,293 Chinese middle-aged and older adults aging over 45 were included at baseline 2011 in this study, and 3,932 participants who accomplished all 4 interviews from 2011 to 2018 were adapted for longitudinal analyses. Sensory status and subjective wellbeing measurements were collected. Other covariates included socio-demographic characteristics, medical condition and lifestyle-related factors. The impacts of baseline sensory status on LE, LS and SRH were assessed using univariate and multivariate logistic regression analyses. A linear regression analysis with generalized estimating equations (GEE) was used to assess the association between time-varying sensory statuses with LE, LS and SRH over 8 years after being adjusted with multi-confounding factors. Results: Participants with SIs had significantly lower level of LE, LS, and SRH, compared to those who were free of SI. All kinds of SIs were significantly associated with LE, LS, and SRH according to cross-sectional data. The correlations between SIs and LE or SRH over 8 years were also noticed. However, only SHI and DSI were found to be significantly associated with LS according to longitudinal data (all p values < 0.05). Conclusion: Sensory impairments had explicitly detrimental effects on subjective wellbeing status over time among middle-aged and older Chinese population.
Assuntos
Surdocegueira , População do Leste Asiático , Perda Auditiva , Transtornos da Visão , Idoso , Humanos , Pessoa de Meia-Idade , Envelhecimento , Estudos Transversais , Estudos Longitudinais , Inquéritos e QuestionáriosRESUMO
BACKGROUND: In a previous hospital-based study, conjunctivochalasis was diagnosed in 85.24% of 1416 patients aged 1 to 94 years and in 98.5% of patients aged 60 or older. This report is the first to present data from a large scale epidemiologic study on conjunctivochalasis in a community-based population, thereby helping to better identify the severity of conjunctivochalasis in the general population. METHODS: This community-based epidemiologic study was conducted to evaluate prevalence rates and related factors of conjunctivochalasis among people over 60 years old in the CaoYangXinCun community, Shanghai, China. Cluster sampling was used in randomly selected local residents aged no less than 60 years. A cross-sectional study using a slit-lamp ophthalmoscope for examination was carried out from September 2008 to October 2008. A modified grading system based on the well-accepted Meller and Tseng's system was used. SPSS10.0 software was used for data and statistical analysis. RESULTS: A total of 2110 residents took part in this study, with a response rate of 94.85%. Among these, 930 cases were confirmed as conjunctivochalasis, with a prevalence rate of 44.08%. The prevalence rate increased with age (X2 = 10.44, P < 0.01). A total of 1762 eyes were confirmed as conjunctivochalasis eyes. Of these eyes, 943 were classified as Grade I (53.52%), 647 as Grade II (36.72%), 162 as Grade III (9.19%), and the remaining 10 eyes as Grade IV (0.57%). The conjunctiva usually accumulated on the nasal and temporal areas of the conjunctival sac (944 eyes, 53.58%). CONCLUSIONS: Although the rate estimated in the present study is not as high as reported in the former hospital-based study, conjunctivochalasis is considered a common age-related eye disease, deserving more attention to its early diagnosis.
Assuntos
Doenças da Túnica Conjuntiva/epidemiologia , Idoso , Idoso de 80 Anos ou mais , Envelhecimento/fisiologia , China/epidemiologia , Doenças da Túnica Conjuntiva/classificação , Estudos Transversais , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Prevalência , Fatores de Risco , Índice de Gravidade de DoençaRESUMO
OBJECTIVE: This study aimed to demonstrate the state of the present situation and trends concerning the global use of acupuncture for cancer pain in the past 20 years. METHODS: Searched the Web of Science database from 2000 to 2019 related to acupuncture for cancer pain, and then used CiteSpace to conduct scientometric analysis to acquire the knowledge mapping. RESULTS: Yearly output has increased year by year, and the growth rate has become faster after 2012. According to the cluster analysis of institutions, authors, cited references, and keywords, 4, 4, 15, and 14 categories were obtained, respectively. The most productive countries, institutions, and authors are the USA, Mem Sloan Kettering Cancer Center, and Mao JJ, whose frequencies are 196, 24, and 17, respectively. However, the most important of them are Australia, Univ. Maryland, and Bao T, owing to their highest centrality, they are 0.90, 0.21, and 0.09 separately. Moreover, cited references that contributed to the most co-citations are Crew KD (2010), however, the most key cited reference is Roscoe JA (2003). Keywords such as acupuncture, pain, breast cancer, palliative care, and quality of life are the most frequently used. But auricular acupuncture is the crucial keyword. In the cluster analysis of institutions, authors, cited references, and keywords, the more convincing research categories are multiple myeloma, placebo effect, neck malignancies, and early breast cancer, with S values of 0.990, 0.991, 0.990, and 0.923, respectively. Therefore, they can be regarded as research hotspots in this field. CONCLUSION: Based on the scientometric analysis in the past 20 years, the knowledge mapping of the country, institution, author, cited reference, and the keyword is gained, which has an important guiding significance for quickly and accurately positioning the trend in this field.
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OBJECTIVE: To analysis tear proteins by using shotgun strategy, and to study pathogenesis mechanism of conjunctivochalasis by comparing tear proteins between the conjunctivochalasis and normal body. METHODS: Tears were obtained from 8 conjunctivochalasis cases and 8 normal controls. Fifteen microliters of tears was collected by microcapillary tubes from each eye. Shotgun strategy was used for tear protein analysis. Trypsin digestion in-solution, separation of peptide mixture by reverse-phase high-pressure liquid chromatograph (RP-HPLC) followed by electrospray ionization mass spectrometry (ESI-MS/MS) identification and bioinformation analysis were used in this study. RESULTS: Three hundred and fifty-six proteins were excised in conjunctivochalasis patients and 352 proteins in the normal controls, and among them 119 proteins were the same. The functions of tear proteins were classified with GOA, which found some apoptosis regulation proteins and apoptosis related proteins and inflammatory response related proteins in conjuntivochalasis but not in normal controls group. Defensin was also found in conjuntivochalasis. CONCLUSION: Shotgun strategy can separate and analysis tear proteins effectively, which provide a new method for tear protein component of conjunctivochalasis. The special component in conjunctivochalasis tear show conjunctivochalasis maybe related to cell apoptosis and inflammatory.
Assuntos
Doenças da Túnica Conjuntiva/metabolismo , Proteínas do Olho/metabolismo , Proteômica , Lágrimas/metabolismo , Idoso , Idoso de 80 Anos ou mais , Estudos de Casos e Controles , Feminino , Humanos , MasculinoRESUMO
OBJECTIVE: To investigate the prevalence and relative factors of conjunctivochalasis among people over 60 years old in Caoyangxincun community of Shanghai. METHODS: A cross-sectional study based on local residents in this community was carried out from September 2008 to October 2008. Cluster sampling was used in randomly selecting individuals aged > or = 60 years. Slit-lamp ophthalmoscope was used for eye examination and diagnosis. The survey was preceded by a pilot study where operational methods were refined and quality assurance evaluation was carried out. A database was set using SPSS 10.0 software, Chi-square test was used for data analysis. RESULTS: 2110 residents took part in this study, with the response rate 94.85%. 930 cases (1762 eyes) were confirmed as conjunctivochalasis, so the prevalence rate is 44.08%. The prevalence of conjunctivochalasis was increasing with age (chi(2) = 10.44, P < 0.01). 556 cases (943 eyes) were confirmed as degree I, 246 cases (647 eyes) as degree II, 120 cases (162 eyes) as degree III and the rest 8 cases (10 eyes) as degree IV. Several symptoms, such as blurred vision, asthenopia and eye pain were more frequent in patient group than in the normal ones (chi(2) value was 15.44, 20.54 and 19.74, respectively, P < 0.01). In the disease eyes, the conjunctiva usually piled up on the nasal and temporal side (944 eyes, 53.58%), the location of inferior lid margin was usually above corneal limbus (2589 eyes, 61.35%) and inferior lid margin entropion and introversion were always found (7.04% and 6.63%). Abnormal lacrimal river was found in 87.23% of the diseased eyes, more than the non-conjunctivochalasis eyes (chi(2) = 1615.81, P < 0.01). CONCLUSIONS: Conjunctivochalasis is a common eye disease in aged population, with obvious signs and symptoms of ocular surface and lacrimal river impairment. In the local residents, most patients were classified as minimal or moderate degree. The location of inferior lid margin and the tension of inferior lid may be related factor of conjunctivochalasis.
Assuntos
Doenças da Túnica Conjuntiva/epidemiologia , Idoso , Idoso de 80 Anos ou mais , China/epidemiologia , Estudos Transversais , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , PrevalênciaRESUMO
Objective: To investigate the treatment effects of botulinum toxin-A (BTA) injection and acupuncture on blepharospasm (BP) evaluated by the change in lower eyelid tension (LET). Methods: A series of 30 patients (male: 8, female: 22) aged between 37 and 83 years (63.80 ± 10.96 yrs) who met the eligibility criteria of BP were recruited in this study, who were randomly assigned to BTA injection group (BTA group, n = 15) and acupuncture treatment group (Acupuncture group, n = 15). BTA injections were administered to the patients in BTA group while patients in acupuncture group received the acupuncture treatment. The LET was measured by a tensiometer in both groups at baseline and at post-treatment. Results: A significant decrease in LETs over 8 weeks was found in acupuncture group (812.76 ± 193.95 Pa at baseline, 549.69 ± 150.04 Pa at 4 weeks, and 510.96 ± 150.66 Pa at 8weeks, respectively; F = 31.127, p << 0.001). There was a significant decrease in LET from 858.61 ± 190.54 Pa at baseline to 414.45 ± 63.38 Pa at 2 weeks after treatment (Z = -4.542, p << 0.01) in BTA group. At the endpoint of the study, a significant difference in LET was found between the acupuncture group (301.80 ± 181.77 Pa) and the BTA group (444.16 ± 193.44 Pa) (t = -2.077, p = 0.047). Conclusions: BP patients have an increased LET. Both BTA and acupuncture are effective in decreasing the LET. Close monitoring of LET holds promise in planning the treatment strategy for Blepharospasm.
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Terapia por Acupuntura , Blefarospasmo/terapia , Toxinas Botulínicas Tipo A/administração & dosagem , Pálpebras/fisiologia , Fármacos Neuromusculares/administração & dosagem , Adulto , Idoso , Idoso de 80 Anos ou mais , Blefarospasmo/tratamento farmacológico , Blefarospasmo/fisiopatologia , Feminino , Humanos , Injeções Intramusculares , Masculino , Pessoa de Meia-Idade , Músculos Oculomotores/efeitos dos fármacosRESUMO
Purpose: Conjunctivochalasis (CCH) is a common ocular disease and has received extensive attention recently. However, its exact pathogenesis remains largely unknown. Owing to the high morbidity of CCH in older people, this study aimed to investigate whether cellular senescence contributes to CCH progression and the underlying mechanism. Methods: Loose conjunctival tissues from CCH patients (n = 13) and normal conjunctival tissues from age-matched persons (n = 12) were obtained and the fibroblasts were separately induced and obtained. Cellular senescence, and the expression of senescence-associated genes (p53 and p21) and p38 in CCH conjunctival tissues and normal controls, were determined by senescence-associated ß-galactosidase (SA-ß-Gal) staining and quantitative (q)RT-PCR, respectively. To explore the effects of p38 on cellular senescence in CCH fibroblasts, small interfering RNA (siRNA) targeting p38 (siP38) and p38-specific inhibitor SB203580 was performed in CCH fibroblasts. Then, cellular senescence, cell viability, reactive oxygen species (ROS) production, and gene expression were detected according to the corresponding methods. Results: CCH conjunctival tissues had significantly more senescent cells, evidenced by more SA-ß-Gal-positive cells, and higher expression of senescence-associated genes (p53 and p21) and p38. CCH fibroblasts transfected with siP38 or treated with SB203580 had obviously reduced numbers of senescent cells, decreased ROS production, and increased cell viability, as well as reduced expression of senescence-associated genes. Meanwhile, blocking p38 signaling decreased the expression of p53 and p21. Conclusions: Therefore, these findings indicate that cellular senescence might be a causative factor for CCH. P38 signaling might play an important role in the progress of cellular senescence in CCH fibroblasts via manipulation of p53/p21 signaling.
Assuntos
Senescência Celular/fisiologia , Doenças da Túnica Conjuntiva/enzimologia , Fibroblastos/enzimologia , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo , Western Blotting , Contagem de Células , Proliferação de Células , Células Cultivadas , Doenças da Túnica Conjuntiva/patologia , Inibidores Enzimáticos/farmacologia , Fibroblastos/efeitos dos fármacos , Humanos , Imidazóis/farmacologia , Piridinas/farmacologia , RNA Interferente Pequeno/farmacologia , Espécies Reativas de Oxigênio/metabolismo , Reação em Cadeia da Polimerase em Tempo Real , Transdução de Sinais , Superóxido Dismutase/metabolismo , beta-Galactosidase/metabolismoRESUMO
This study investigated the potential role of MAPK signaling pathways in conjunctivochalasis (CCH). Twenty loose conjunctival biopsy samples from 20 CCH and 15 conjunctival biopsy samples from 15 normal controls (CON) were collected. The conjunctival fibroblasts were cultured in vitro. Immunofluorescence, ELISA, Western blot and reverse transcription-polymerase chain reaction (RT-PCR) were used. Our results showed that the expression of p-ERK, p-JNK, and p-p38 in CCH conjunctiva was significantly higher than that in CON group. The expression of p38 MAPK, JNK, and ERK proteins in CCH fibroblasts was significantly higher than that in CON group. The total expression of MAPK mRNA in CCH fibroblasts was significantly higher than that in CON group. The activated forms of p38 MAPK, JNK, and ERK proteins and mRNAs might up-regulate the expression of MMPs in CCH loose conjunctival tissue and fibroblasts, causing the degradation of collagen fibers and elastic fibers and promoting the occurrence of CCH. Our results deepen the understanding of CCH pathological mechanism.
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BACKGROUND: With the aging of the population and the use of video terminals, the incidence of Conjunctivochalasis is getting higher, and related research is increasing. So our research aimed to use visualization software to display the research trends of Conjunctivochalasis. METHODS: Retrieved the document (from 1986 to 2017) of conjunctivochalasis in the web of science core collection, analyzed by Citespace V. RESULTS: The main language is English. Article is the key type of document. The average annual number of publications in the time period from 2008 to 2017 was 11.6, which was significantly higher than the period from 1994 to 2007, indicating that the total number of publications has been continuously developed. The law of frequency quoted showed an upward trend yearly. Furthermore, we can find out that Japan, USA, and People's Republic of China were the most productive countries, Kyoto Prefectural University of Medicine was the most prolific institution, Shanghai Jiaotong University is a key institution. The average IF of journals was 3.0508. Cornea and Canadian Journal of Ophthalmology are core journals. Tseng SCG is the most active scholar. All cited author contributed to 5 classifications. Di PMA paper is a classic literature. Huang YK paper can be regarded as the frontier document. All cited-reference dedicated to 7 categories. Conjunctivochalasis is the hot topic, related to observe indicators, risk factors, treatment, graded diagnosis of conjunctivochalasis, etc. In addition, fibroblast was research hotspot. At length, the cluster map of keyword was divided into 7 categories. CONCLUSION: This research will help relevant clinicians and researchers to accurately and quickly grasp the research trends in the field, and continue to conduct new research on the basis.
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Bibliometria , Pesquisa Biomédica/tendências , Doenças da Túnica Conjuntiva , Humanos , Editoração/tendências , SoftwareRESUMO
Proteins in the aqueous humor (AH) are important in the induction of cataract development. The identification of cataract-associated proteins assists in identifying patients and predisposed to the condition and improve treatment efficacy. Proteomics analysis has previously been used for identifying protein markers associated with eye diseases; however, few studies have examined the proteomic alterations in cataract development due to high myopia, glaucoma and diabetes. The present study, using the isobaric tagging for relative and absolute protein quantification methodology, aimed to examine cataract-associated proteins in the AH from patients with high myopia, glaucoma or diabetes, and controls. The results revealed that 445 proteins were identified in the AH groups, compared with the control groups, and 146, 264 and 130 proteins were differentially expressed in the three groups of patients, respectively. In addition, 44 of these proteins were determined to be cataractassociated, and the alterations of five randomly selected proteins were confirmed using enzyme-linked immunosorbent assays. The biological functions of these 44 cataract-associated proteins were analyzed using Gen Ontology/pathways annotation, in addition to proteinprotein interaction network analysis. The results aimed to expand current knowledge of the pathophysiologic characteristics of cataract development and provided a panel of candidates for biomarkers of the disease, which may assist in further diagnosis and the monitoring of cataract development.
Assuntos
Humor Aquoso/química , Catarata/metabolismo , Proteínas do Olho/isolamento & purificação , Idoso , Idoso de 80 Anos ou mais , Humor Aquoso/metabolismo , Biomarcadores/análise , Biomarcadores/metabolismo , Catarata/diagnóstico , Catarata/genética , Diabetes Mellitus/metabolismo , Proteínas do Olho/genética , Proteínas do Olho/metabolismo , Feminino , Glaucoma/metabolismo , Humanos , Masculino , Pessoa de Meia-Idade , Miopia/metabolismo , Proteômica/instrumentação , Proteômica/métodosRESUMO
AIM: To investigate the expression of matrix metalloproteinases 1 and 3 (MMP-1 and MMP-3) and their tissue inhibitors of metalloproteinases 1 and 3 (TIMP-1 and TIMP-3) in the conjunctiva of eyes with conjunctivochalasis (CCh). METHODS: The conjunctival tissue was obtained from the CCh patients and controls, the MMPs/TIMPs expression concentration was determined by enzyme-linked immuno-sorbent assay (ELISA) and immunofluorescence staining. The expression levels of MMPs/TIMPs in the CCh fibro-blasts were determined by analyzing its concentration in the cellular supernatant that was abstracted from the in vitro cultured CCh fibroblasts. RESULTS: MMP-1 and MMP-3 levels determined by ELISA were both significantly higher in the CCh group than that in the control group (P=0.042, 0.022, respectively), so was the levels of TIMP-1 (P=0.010). No significant difference in the expression of TIMP-3 in conjunctiva was found between the two groups (P=0.298). The expression of MMP-1 and MMP-3 were both up-regulated significantly in the CCh group (P=0.040, 0.001, respectively) on immuno-fluorescence staining. MMP-1 and MMP-3 expression in the fibroblasts were both significantly higher in the CCh group than that in the control group (P=0.027, 0.001, respectively), while neither the TIMP-1 nor TIMP-3 expression was significantly different between the two groups (P=0.421, 0.237, respectively). CONCLUSION: The overexpression of MMP-1 and MMP-3 in conjunctival tissue and fibroblasts may play an important role in the pathogenesis and development of CCh.
RESUMO
PURPOSE: Eyelid tension seems to be related to corneal astigmatism and to affect the ocular surface. The aim of this study is to determine the lower eyelid tension in young adults with a simple lid tensiometer. METHODS: A commercially available precision digital pressure gauge that was connected to a pressure-guided tube full of water with a sensor at its end being placed between the lower eyelid and ocular surface was used as the lid tensiometer to measure the lower eyelid pressure at the central lid in 8 male and 12 female subjects aged between 20 and 39 years with normal healthy eyes. Measurements were respectively performed by 2 operators under the same conditions to test possible interoperator variation. RESULTS: The lower eyelid pressures of the 20 subjects measured by 2 operators at the central lid were 445.28 ± 121.17 and 458.65 ± 127.15 Pa, respectively. The test of interoperator variation demonstrated that there was good agreement between 2 operators (intraclass correlation coefficient = 0.965, F = 56.09, P < 0.001). CONCLUSIONS: Our simple lid tensiometer is a viable option for measuring eyelid pressure with good repeatability.
Assuntos
Técnicas de Diagnóstico Oftalmológico/instrumentação , Pálpebras/fisiologia , Tono Muscular/fisiologia , Músculos Oculomotores/fisiologia , Pressão , Adulto , Feminino , Voluntários Saudáveis , Humanos , Masculino , Manometria/instrumentação , Adulto JovemRESUMO
PURPOSE: To conduct in vivo measurements of the bulbar conjunctiva epithelial, stromal, and total thicknesses in healthy Chinese subjects using optical coherence tomography (OCT) and to compare the conjunctival thickness differences between sex and age. METHODS: From January to July 2012, healthy Chinese subjects were recruited to undergo a Cirrus HD-OCT examination using the Anterior Segment 5 Line Raster scanning protocol. Ophthalmoscopy and slit-lamp examinations were performed on all eyes, along with the determination of visual acuity. The bulbar conjunctiva epithelial thickness, stromal thickness, and full thickness were measured on cross-sectional conjunctiva images. The average values of each parameter were compared between males and females and among eight age groups (<20, 21-30, 31-40, 41-50, 51-60, 61-70, 71-80, and >80 years). RESULTS: A total of 711 eyes in 711 subjects (average age 46.5 years) were included. The average conjunctiva epithelial, stromal, and full thickness measurements were 42.4 ± 7.4 µm, 197.7 ± 32.5 µm, and 240.1 ± 29.8 µm, respectively. No significant difference was found for any of the three parameters between sexes. The conjunctiva epithelial thickness values decreased significantly after age 20 and were then maintained at relatively low levels, followed by a sharp increase at 60 years of age. The conjunctiva stromal thickness decreased throughout the entire lifetime, and the full thickness decreased from the teenage to the elderly periods. CONCLUSIONS: None of the Chinese conjunctiva epithelial, stromal, and full thickness was sex-related, but all of them were age-related.