Properties, bioactive potential and extraction processes of glycosaminoglycans: an overview / Propriedades, potencial bioativo e processos de extração de glicosaminoglicanos: uma visão geral

ABSTRACT: Glycosaminoglycans (GAGs) are long-chain polysaccharides that are divided into sulphates and non-sulphates, these being chondroitin sulphate, heparan sulphate, dermatan sulphate, heparin sulphate and the only non-sulphate in the group is hyaluronic acid. GAGs are obtained from animal tissue and by an expensive low-yield extraction process; however, they are highly commercially valued polysaccharides and exploited in the biomedical market. Their disaccharidic composition, chain length and sulfation pattern present great variability depending on the species and extraction factors. GAGs possess immunomodulatory, antioxidant, antiviral, anti-inflammatory, neuroprotective, antiproliferative and anticoagulant properties, functioning as therapeutic agents modulating an array of biological processes. This report presents the general aspects of each GAG, source and extraction process, in addition to the characteristics that give them the most varied therapeutic properties and pharmacological applications.

RESUMO: Os glicosaminoglicanos (GAGs) são polissacarídeos de cadeias longas que se dividem em sulfatados e não sulfatados, sendo estes, sulfato de condroitina, sulfato de heparana, sulfato de dermatana, sulfato de heparina e o único não sulfatado do grupo que é o ácido hialurônico. Os GAGs são encontrados em todo tecido animal, são extraídos por um processo de alto custo e baixo rendimento, no entanto, o material obtido é valorizado comercialmente e amplamente explorado no mercado biomédico. Sua composição dissacarídica, comprimento da cadeia e padrão de sulfatação apresentam grande variabilidade dependendo das espécies e fatores de extração. Os GAGs possuem propriedades imunomoduladoras, antioxidantes, antivirais, anti-inflamatórias, neuroprotetoras, antiproliferativas e anticoagulantes, além de farmacológicas, funcionando como agentes terapêuticos moduladores de uma série de processos biológicos. Este relatório apresenta os aspectos gerais de cada GAG, fonte e processo de extração, além das características que lhes conferem as mais variadas propriedades terapêuticas e aplicações farmacológicas.

Clinical and Radiological Characterisation of Patients with Mucopolysaccharidosis in a Genetic Clinic

Abstract The mucopolysaccharidoses (MPS) are a relatively uncommon group of inherited metabolic disorders. MPSs should be suspected in a child with coarse facies, organomegaly, recurrent respiratory tract infections, developmental delay, and hernias. Early diagnosis and treatment can greatly improve the quality of life in these children. In this study we studied 46 MPS patients diagnosed on enzyme and/or DNA testing and we found that the MPS II was the most common type followed by MPS I and MPS IVA. While the mean age of onset of symptoms was 12 months, the mean age at diagnosis was 4.5 years, a significant delay. One of major presenting features was recurrent respiratory problems, more prevalent in MPS II cases. Many patients also had short stature and contractures. Increasing awareness among physicians is of paramount importance for the early diagnosis and optimal treatment and prevention by prenatal testing and counselling.

Chondroitin sulfate from fish waste exhibits strong intracellular antioxidant potential

Chondroitin sulfate (CS) is a type of glycosaminoglycan described as an antioxidant molecule that has been found in animal species such as fish. Tilapia (Oreochromis niloticus) represents an eco-friendly source of this compound, since its economical processing generates usable waste, reducing the negative environmental impact. This waste was used for CS extraction, purification, characterization by enzymatic degradation, and evaluation of its antioxidant effect. CS obtained from tilapia presented sulfation mainly at carbon 4 of galactosamine, and it was not cytotoxic at concentrations up to 200 µg/mL. Furthermore, 100 µg/mL of CS from tilapia reduced the levels of reactive oxygen species to 47% of the total intracellular reactive oxygen species level. The ability of CS to chelate metal ions in vitro also suggested an ability to react with other pathways that generate oxidative radicals, such as the Haber-Weiss reaction, acting intracellularly in more than one way. Although the role of CS from tilapia remains unclear, the pharmacological effects described herein indicate that CS is a potential molecule for further study of the relationship between the structures and functions of chondroitin sulfates as antioxidants.

Intravesical Glycosaminoglycans Experience for Chronic Lower Urinary Tract Pathology Treatment / Experiencia con glucosaminoglicanos intravesicales para el tratamiento de patologías crónicas del tracto urinario inferior

Objective Intravesical glycosaminoglycans (GAG) treatment is one of the therapeutic options for chronic bladder pathologies and is approved for Bladder Pain Syndrome (BPS), radiation cystitis, and recurrent urinary tract infections (UTIs). The purpose of this study is to describe the demographic characteristics of patients with such pathologies in our population and to evaluate treatment response. Methods It is a retrospective study of patients with the aforementioned pathologies, who received treatment with GAG. Demographic characteristics and subjective improvement with treatment were evaluated. A bivariate analysis was performed to study possible improvement predictors. Results 53 patients were evaluated. Of them, 33 (62.3%) with BPS, 12 (22.6%) with recurrent UTIs, and 8 (15.1%) with radiation cystitis. The dose range received was between 4 - 20 instillations, with a median of 6 doses. 67.9% of patients showed improvement of symptoms with treatment, this percentage being even higher for the group of patients with recurrent UTIs (91%). No treatment response predictors were found. Conclusions Intravesical GAG treatment is a therapeutic alternative for patients with chronic bladder pathologies, with satisfactory results in the medium term. Prospective studies are needed to support the findings of this study.

Objetivo El tratamiento con glucosaminoglicanos intravesicales hace parte de las opciones terapéuticas de las patologías crónicas de la vejiga y se encuentra aprobado para el tratamiento de síndrome de vejiga dolorosa (SVD), cistitis por radiación e infección urinaria recurrente. El objetivo de este estudio es describir las características demográficas de los pacientes con dichas patologías en nuestra población y evaluar la respuesta al tratamiento. Métodos Estudio retrospectivo de pacientes con las patologías mencionadas, quienes recibieron tratamiento con glucosaminoglicanos. Se evaluaron las características demográficas y la respuesta al tratamiento. Se realizó un análisis bivariado para estudiar posibles variables predictoras de mejoría. Resultados Se evaluaron 53 pacientes. De estos, 33 (62.3%) con síndrome de vejiga dolorosa, 12 (22.6%) con infección urinaria recurrente y 8 (15.1%) con cistitis por radiación. El rango de dosis recibida estuvo entre 4-20 instilaciones, con una mediana de 6 dosis. El 67.9% de los pacientes tuvo una respuesta adecuada al tratamiento, siendo este porcentaje aún mayor para el grupo de pacientes con IVU recurrente (91%). No se encontraron factores predictores de respuesta al tratamiento. Conclusiones El tratamiento con GAG intravesicales es una alternativa terapéutica para pacientes con patologías crónicas de la vejiga, con resultados satisfactorios a mediano plazo. Se necesitan estudios prospectivos que soporten los hallazgos de este trabajo.

Case series of patients under biweekly treatment with laronidase: a report of a single center experience / Série de casos de pacientes em tratamento com laronidase quinzenal: relato da experiência de um centro

ABSTRACT Objective: To report the stabilization of urinary glycosaminoglicans (GAG) excretion and clinical improvements in patients with mucopolysaccharidosis type I (MPS I) under an alternative dose regimen of laronidase of 1.2 mg/kg every other week. Methods: We participated in a dose-optimization trial for laronidase in MPS-I patients using four alternative regimens: 0.58 mg/kg every week, 1.2 mg/kg every two weeks, 1.2 mg/kg every week and 1.8 mg/kg every other week (EOW). After the trial ended, the patients resumed the recommended dose and regimen of 0.58 mg/kg every week. Under this regimen, some patients presented difficulties in venous access and were unable to commute weekly to the treatment center. Therefore, we used an alternative regimen that consisted of 1.2 mg/kg EOW in eight patients. A retrospective study of medical records of MPS-I patients who underwent both enzyme replacement therapy (ERT) regimens, of 0.58 mg/kg every week and 1.2 mg/kg EOW, was done. Results: Patients remained clinically stable under the alternative regimen, did not present elevation of urinary GAG nor any adverse event. Conclusions: The switch of dose regimen to 1.2 mg/kg EOW of laronidase was safe, and did not cause any clinical worsening in patients who had been previously under standard dose ERT.

RESUMO Objetivo: Descrever a manutenção dos níveis de glicosaminoglicano (GAG) excretados na urina e da estabilização clínica em pacientes com mucopolissacaridose do tipo I (MPS I) com o uso da laronidase num regime de dose alternativo de 1,2 mg/kg a cada duas semanas. Método: Alguns pacientes do nosso serviço participaram de um estudo de otimização de dose da laronidase para o tratamento da MPS I no qual foram comparados quatro esquemas terapêuticos: 0,58 mg/kg/semana, 1,2 mg/kg a cada duas semanas, 1,2 mg/kg/semana e 1,8 mg/kg a cada duas semanas. Após o término do estudo, todos os pacientes passaram a receber a terapia de reposição enzimática (TRE) na dose padrão de bula, que é de 0,58 mg/kg/semana, e nesse regime alguns pais se queixaram da dificuldade em comparecer ao centro todas as semanas, além da dificuldade de se obter acesso para punção venosa. Com base nessas queixas, oito pacientes passaram a receber a TRE no regime alternativo de 1,2 mg/kg a cada duas semanas. Foi feito o estudo retrospectivo de dados de prontuário de pacientes com MPS I que fizeram TRE com laronidase nas doses 0,58 mg/kg/semana e 1,2 mg/kg a cada duas semanas. Resultados: Os pacientes mantiveram-se clinicamente estáveis, não apresentaram aumento dos níveis de GAG urinários nem eventos adversos durante o regime alternativo de dose. Conclusões: A mudança para o esquema de 1,2 mg/kg de laronidase a cada duas semanas foi segura e não acarretou piora clínica nos pacientes que já estavam em TRE na dose padrão.

Macrophage migration inhibitory factor antagonist (p425) ameliorates kidney histopathological and functional changes in diabetic rats / Antagonista (p425) do fator de inibição da migração de macrófagos (MIF) melhora as alterações histopatológicas e funcionais renais em ratos diabéticos

Abstract Introduction: It is hypothesized that increased macrophage migration inhibitory factor (MIF) expression may contribute to diabetic nephropathy (DN) pathogenesis. The aim of the present study was to investigate the renal effects of MIF inhibition in a diabetic experimental model. Methods: Eighteen male Wistar rats (230 ± 20 g) were divided into three groups: 1) control, 2) diabetic (STZ, 50 mg/kg, dissolved in saline, ip), 3) diabetic + MIF antagonist (p425, 1 mg/kg per day, ip, on the 21th day, for 21 consecutive days). The treatment started since we founwd a significant increase in urine albumin excretion (UAE) rate in the diabetic rats in comparison with the control rats. The rats were kept individually in metabolic cages (8 AM-2 PM) and urine samples were collected in the 21 and 42th day. At the end, blood and tissue samples were collected for biochemical (BS, UPE, urine GAG, BUN, Cr, Na, and K) and histological analyses. Results: The results of this study showed that MIF antagonist (p425) significantly decreased urine protein and GAG excretion, urine protein/creatinine ratio, and serum BUN and Cr in the streptozotocin-induced DN in the rats. Pathological changes were significantly alleviated in the MIF antagonist (p425)-administered DN rats. Conclusion: Collectively, these data suggested that MIF antagonist (p425) was able to protect against functional and histopathological injury in the DN.

Resumo Introdução: Supõe-se que elevações da expressão do fator de inibição da migração de macrófagos (MIF) possam contribuir para a patogênese da nefropatia diabética (ND). O objetivo do presente estudo foi investigar os efeitos renais da inibição do MIF em um modelo experimental diabético. Métodos: Dezoito ratos Wistar machos (230 ± 20g) foram divididos em três grupos: 1) controle, 2) diabético (STZ 50 mg/kg dissolvida em soro fisiológico, IP), 3) diabético + antagonista do MIF (p425 1 mg/kg por dia IP no 21o dia por 21 dias consecutivos). O tratamento começou após a identificação de aumento significativo na albuminúria nos ratos diabéticos em relação aos controles. Os ratos foram mantidos individualmente em gaiolas metabólicas (8h-14h) e amostras de urina foram colhidas no 21o e no 42o dia. Ao final do estudo, amostras de sangue e tecido foram colhidas para análises bioquímicas (BS, excreção urinária de proteína, excreção urinária de GAGs, BUN, Cr, Na e K) e histológicas. Resultados: O presente estudo demonstrou que o antagonista do MIF (p425) diminuiu significativamente proteinúria, excreção urinária de GAGs , relação proteína/creatinina na urina, BUN e Cr no grupo com ND induzida por estreptozotocina. As alterações patológicas foram significativamente abrandadas nos ratos com ND que receberam antagonista do MIF (p425). Conclusão: Coletivamente, os dados sugerem que o antagonista do MIF (p425) teve efeito protetor contra lesões funcionais e histopatológicas da ND.

Lichen myxedematosus: a rare group of cutaneous mucinosis

Abstract: Cutaneous mucinoses are a heterogeneous group of dermatoses in which excess deposition of mucin in the dermis gives the skin a waxy appearance, with papules and plaques that can vary from self-healing mucinosis to even disrupting the normal shape of a patient's face, conferring a leonine facies, or be part of life threatening diseases like scleromyxedema. This review will describe the most recent classification on lichen myxedematosus in the generalized (scleromyxedema) and the localized forms, as well as the different organ systems involved in scleromyxedema, diagnostic workup, current management, and prognosis.

Daño oxidativo y capacidad de defensa antioxidante en pacientes cubanos con mucopolisacaridosis / Oxidative damage and anti-oxidant defense capacity in Cuban patients with mucopolysaccharidosis

Introducción: Las mucopolisacaridosis son causadas por la deficiencia de las actividades de las enzimas lisosomales necesarias para degradar los glicosaminoglicanos. Estos síndromes comparten muchas características clínicas aunque en grados variables. Las manifestaciones clínicas implican múltiples sistemas de órganos y algunas tienen terapia de reemplazo enzimático. En muchas investigaciones se hace alusión a la presencia de estrés oxidativo en quienes la padecen, pero esta condición aún no se ha estudiado en los pacientes cubanos. Objetivo: Evaluar parámetros de estrés oxidativo en pacientes cubanos con mucopolisacaridosis. Métodos: Se realizó un estudio de casos y controles que incluyó a 7 niños con mucopolisacaridosis de tipos I, II, III y IV (casos) y a 21 aparentemente sanos, pareados en edad y sexo (controles). Se midieron los niveles plasmáticos de malonildialdehído, productos avanzados de oxidación de proteínas, grupos tiol libres y marcadores de química sanguínea. Se cuantificaron las actividades intraeritrocíticas de superóxido dismutasa, catalasa y de glutatión peroxidasa. Todas las técnicas utilizadas fueron espectrofotométricas. Resultados: Los pacientes mostraron un aumento tanto en los niveles de calcio como en la oxidación de lípidos y proteínas, en comparación con los controles y los valores de referencia de Cuba. Hubo una disminución en la actividad de la enzima superóxido dismutasa y las concentraciones de grupos tioles. No se encontraron diferencias para el resto de los parámetros medidos. Conclusiones: El aumento del daño oxidativo y la disminución de la capacidad antioxidante sugieren la presencia de estrés oxidativo en esos pacientes cubanos.

Introduction: Mucopolysaccharidosis are caused by the deficiency in lysosomal enzyme activities necessary to degrade the glycosaminoglycans. These syndromes share many clinical characteristics although in variable degrees. Clinical manifestations imply multiple organs systems and some have enzyme replacement treatment. Many investigations deal on the presence of oxidative stress in those who suffer it, but this condition has not still been studied in Cuban patients. Objective: To evaluate parameters of oxidative stress in Cuban patients with mucopolysaccharidosis. Methods: A cases and controls study which included 7 children with mucopolysaccharidosis types I, II, III and IV (cases) and 21 apparently healthy children, paired by age and sex (control group) was carried out. The plasmatic levels of malondialdehide, advanced products of proteins oxidation, free thiol groups and blood chemistry markers were measured. The intraerythrocytic activities of superoxide dismutase, catalase and that of glutathione peroxidase were quantified. All the used techniques were spectrophotometrical. Results: The patients showed an increase, both in the calcium levels as in the oxidation of proteins and lipids, in comparison with the control group and the Cuban values reference. There was a decrease in the activity of the enzyme superoxide dismutase and the concentrations of thiols groups. There were no differences for the rest of the measured parameters. Conclusions: The increase of the oxidative damage and the decrease of the anti-oxidant capacity suggest the presence of oxidative stress in those Cuban patients.

Characterization of Decellularized Human Pericardium for Tissue Engineering and Regenerative Medicine Applications / Caracterização do Pericárdio Humano Descelularizado para Engenharia de Tecidos e Aplicações de Medicina Regenerativa

Abstract Background: Pericardium tissue allograft can be used for surgical repair in several procedures. One of the tissue engineering strategies is the process of decellularization. This process decreases immunogenic response, but it may modify the natural extracellular matrix composition and behavior. Objective: The aim of this study was to evaluate the effectiveness of cell removal, maintenance of extracellular matrix properties and mechanical integrity of decellularized human pericardium using a low concentration solution of sodium dodecyl sulfate. Methods: Decellularization was performed with sodium dodecyl sulfate and ethylenediaminetetraacetic acid. Histological analysis, DNA quantification, evaluation of glycosaminoglycans and collagen were performed. Biomechanical assay was performed using tensile test to compare the decellularization effects on tissue properties of tensile strength, elongation and elastic modulus. P < 0.05 was considered significant. Results: There was reduction in visible nuclei present in pericardium tissue after decellularization, but it retained collagen and elastin bundles similar to fresh pericardium. The DNA contents of the decellularized pericardium were significantly reduced to less than 511.23 ± 120.4 ng per mg of dry weight (p < 0.001). The biomechanical assay showed no significant difference for fresh or decellularized tissue. Conclusion: The decellularization process reduces cell content as well as extracellular matrix components without changing its biomechanical properties.

Resumo Fundameto: O enxerto de pericárdio pode ser usado em muitos procedimentos de correção cirúrgica. Uma das estratégias da engenharia tecidual é o processo de descelularização. No entanto, embora esse processo diminua a resposta imunogênica, a descelularização pode modificar tanto o comportamento como a composição da matriz extracelular natural. Objetivos: Avaliar a eficácia da descelularização usando baixa concentração de dodecil sulfato de sódio na remoção celular, na manutenção das propriedades da matriz extracelular e na integridade mecânica do pericárdio humano descelularizado. Métodos: A descelularização foi realizada com dodecil sulfato de sódio e ácido etilenodiamino tetra-acético. Foi realizada análise histológica, quantificação de DNA, e avaliação de glicosaminoglicanos e colágeno. O estudo biomecânico foi conduzido pelo teste de tração para comparar os efeitos da descelularização sobre as propriedades teciduais de resistência à tração, alongamento e módulo de elasticidade. Foi considerado um valor de p < 0,05 como estatisticamente significativo. Resultados: Observou-se uma redução na quantidade de núcleos presentes no pericárdio após a descelularização, apesar de manter quantidades similares de feixes de elastina e de colágeno. As concentrações de DNA do pericárdio descelularizado foram significativamente reduzidas para menos que 511,23 ± 120,4 ng por mg de peso seco (p < 0,001). O teste biomecânico não apontou diferenças entre os tecidos fresco e descelularizado. Conclusão: A descelularização reduziu a concentração de células bem como os componentes da matriz extracelular sem afetar suas propriedades biomecânicas.

Revisión Bibliográfica: Glicocálix Endotelial como Blanco de Agresión del Fenómeno Isquemia Reperfusión / Bibliographic Review: Endothelial Glycocalyx as Target of Aggression of the Ischemia-Reperfusion Injury

El glicocálix endotelial es una estructura rica en glucosaminoglicanos, proteoglicanos y glucoproteínas que recubre el endotelio vascular; además de ser una estructura de protección, al estar en contacto directo con la sangre se convierte en el blanco de agresión de diversos mecanismos fisiopatológicos. El fenómeno isquemia-reperfusión se presenta comúnmente en varias entidades del paciente crítico, incluyendo: eventos cerebro vasculares isquémicos, síndrome coronario agudo, sepsis y choque en sus distintos tipos, traumatismos mayores, cirugía y trasplante. Las complicaciones derivadas de este fenómeno son múltiples y dependientes del sitio de presentación; el común denominador es la disfunción microvascular que potencialmente podría desencadenar un fallo multisistémico. El objetivo de esta revisión bibliográfica fue realizar una actualización de los conocimientos en relación a la injuria del glicocálix endotelial durante el fenómeno isquemia-reperfusión.(au)

The endothelial glycocalyx is a structure rich in glycosaminoglycans, proteoglycans and glycoproteins that cover vascular endothelium; in addition of being a protective structure, the direct contact with blood turns it the target of aggression of multiple physiopathological mechanisms. The ischemia-reperfusion injury commonly presents in several critical care entities, including: ischemic stroke, acute coronary syndrome, sepsis and shock, major trauma, surgery and transplantation. Complications are multiple and dependent of the site of presentation; the common denominator is microvascular dysfunction that could potentially trigger multiple organ dysfunction syndrome. The aim of this bibliographic review was to update the knowledge regarding endothelial glycocalyx damage and ischemia-reperfusion injury.(au)

Inflammatory biomarkers in sera of patients with intervertebral disc degeneration / Biomarcadores inflamatórios no plasma de pacientes com degeneração do disco intervertebral

ABSTRACT Objective: To evaluate intervertebral disc levels of inflammatory factor (interleukin 6) and proteinase activity (cathepsin B) in patients with a degenerative disease and serum levels of interleukin 6, serum cathepsin B activity and hyaluronic acid biomarkers. Methods: We conducted immunohistochemistry studies of intervertebral discs to analyze interleukin 6 and cathepsin B levels of patients with degenerative disease and spine fracture (Control Group) and to measure hyaluronic acid, interleukin 6 and cathepsin B activity from sera of intervertebral disc degeneration patients, fracture patients, and healthy individuals. Results: Interleukin 6 and cathepsin B seem to be related with physiopathology of intervertebral disc degeneration, since the levels of both were higher in discs of patients with intervertebral disc degeneration. Interleukin 6 and cathepsin B do not represent good biomarkers of degenerative intervertebral disc disease, since the level of such compounds is increased in the plasma of patients with fractures. Conclusion: Hyaluronic acid can be a biomarker for intervertebral disc degeneration, because hyaluronic acid levels were higher only in sera of patients with intervertebral disc degeneration.

RESUMO Objetivo: Avaliar os níveis de fatores inflamatórios nos discos intervertebrais (interleucina 6) e proteinase (catepsina B) em pacientes com doença degenerativa de disco intervertebral, além de verificar os níveis séricos de interleucina 6, ácido hialurônico e atividade sérica da catepsina B. Métodos: Foi realizado exame imuno-histoquímica dos discos intervertebrais de pacientes com doença degenerativa e fratura da coluna (Grupo Controle) e análise do plasma de pacientes com doença degenerativa de disco intervertebral. Como controle, foram utilizados plasma de pacientes com fraturas, além de indivíduos saudáveis. Resultados: Interleucina 6 e catepsina B sugerem relação com a fisiopatologia da doença degenerativa de disco intervertebral, uma vez que os níveis de ambos foram maiores nos discos de pacientes com doença degenerativa de disco intervertebral. Interleucina 6 e catepsina B não representam bons biomarcadores da doença degenerativa do disco intervertebral, já que também encontram níveis aumentados em plasma de pacientes com fratura. Conclusão: O ácido hialurônico é um possível biomarcador de doença degenerativa de disco intervertebral, porque os níveis de ácido hialurônico foram maiores apenas em plasma de pacientes com doença degenerativa de disco intervertebral.

Observation on the efficacy and mechanism of Hirudoid in preventing psoriasis-like dermatitis in mice / 中国医师杂志

Objective To observe the preventive effect of Hirudoid (MPS) ointment on psoriasislike dermatitis in mice and explore its mechanism.Metbods 21 Balb/C mice were randomly divided into three groups:blank control group,positive control group (imiquimod group) and treatment group (imiquimod + Hirudoid group).The blank control group was treated with imiquimod matrix.The positive control group was treated with 5％ imiquimod cream to induce psoriatic dermatitis in mice's back.The treatment group was treated with 5％ imiquimod cream in the morning and Hirudoid cream in the afternoon.The changes of skin erythema,scales,infiltration severity and skin histopathology in the experimental area of mice back and serum levels of interleukin (IL)-17A and protein expression of tumor necrosis factor-α (TNF-α)in the three groups of mice were observed.Results The clinical and pathological findings of skin lesions in the positive control group showed that 5％ imiquimod cream successfully induced psoriasis-like dermatitis in mice.The appearance and pathological feature of skin lesions in the blank control group were almost close to normal skin.The degree of erythema,infiltration and scales in the back of the treatment group was significantly less than that in the positive control group,and the degree of microscopic inflammation in the treatment group was also lighter than that in the positive control group;The serum levels of IL-17A and TNF-α in the treatment group and the blank control group were significantly lower than that of the positive control group,with statistically significant difference (P ＜ 0.05).Conclusions Hirudoid can significantly reduce imiquimod-induced psoriasis-like dermatitis by inhibiting the expression of IL-17A and TNF-α.

Prospective evaluation of Chondroitin sulfate, Heparan sulfate and Hyaluronic acid in prostate cancer

ABSTRACT Purpose: The present study evaluates chondroitin sulfate (CS) and heparan sulfate (HS) in the urine and hyaluronic acid (HA) in the plasma of patients with prostate cancer before and after treatment compared to a control group. Materials and Methods: Plasma samples were used for HA dosage and urine for quantification of CS and HS from forty-four cancer patients and fourteen controls. Clinical, laboratory and radiological information were correlated with glycosaminoglycan quantification by statistical analysis. Results: Serum HA was significantly increased in cancer patients (39.68 ± 30.00 ng/ mL) compared to control group (15.04 ± 7.11 ng/mL; p=0.004) and was further increased in high-risk prostate cancer patients when compared to lower risk patients (p = 0.0214). Also, surgically treated individuals had a significant decrease in seric levels of heparan sulfate after surgical treatment, 31.05 ± 21.01 μg/mL (before surgery) and 23.14 ± 11.1 μg/mL (after surgery; p=0.029). There was no difference in the urinary CS and HS between prostate cancer patients and control group. Urinary CS in cancer patients was 27.32 ± 25.99 μg/mg creatinine while in the men unaffected by cancer it was 31.37 ± 28.37 μg/mg creatinine (p=0.4768). Urinary HS was 39.58 ± 32.81 μg/ mg creatinine and 35.29 ± 28.11 μg/mg creatinine, respectively, in cancer patients and control group (p=0.6252). Conclusions: Serum HA may be a useful biomarker for the diagnosis and prognosis of prostate cancer. However, urinary CS and HS did not altered in the present evaluation. Further studies are necessary to confirm these preliminary findings.

Clinical features and corneal optical anisotropies in a rabbit model of limbal stem cell deficiency / Fenótipos clínicos e anisotropias ópticas da córnea em coelhos com deficiência de células tronco limbais

ABSTRACT Purposes: To investigate the intra-laboratory reproducibility of clinical features and to evaluate corneal optical anisotropies in a rabbit model of limbal stem cell deficiency. Methods: Limbal injury was induced in the right eye of 23 adult New Zealand White rabbits using a highly aggressive protocol that combined 360 degrees limbal peritomy, keratolimbectomy, alkaline chemical burn, and mechanical removal of the epithelium. Clinical evaluation of the injured eyes was performed for 28 days and included corneal impression cytology. Corneas with a severe clinical outcome set typical of limbal stem cell deficiency were then collected, subjected to a histopathological examination, and examined for optical anisotropies. Corneas from healthy rabbit eyes were used as controls. Differences in optical path due to stromal collagen birefringence, as well as linear dichroism related to the expression and spatial orientation of glycosaminoglycan chains from proteoglycans, were measured from cross-sections under a quantitative polarized light microscope. Results: One eye showed signs of hypopyon and was excluded. Signs of ocular inflammation were observed in all eyes studied (n=22). Corneal impression cytology did not detect goblet cells. Twelve of the 22 corneas presented a clinical outcome set typical of limbal stem cell deficiency, which is characterized by the presence of epithelial defects, inflammatory cells, moderate-to-severe opacity, and neovascularization. Microscopic studies under polarized light revealed that relative to controls, limbal stem cell deficiency caused a 24.4% increase in corneal optical path differences. Further, corneas with limbal stem cell deficiency were less dichroic than controls. Conclusions: These results suggest that rabbit models of limbal stem cell deficiency must be rigorously screened for use in preclinical studies to ensure experimental homogeneity because protocols used to create limbal stem cell deficiency could be not associated with good intra-laboratory reproducibility of clinical features. Limbal stem cell deficiency, as induced herein, altered the optical anisotropic properties of the corneal stroma. Such alterations are indicative of changes in collagen packing and the spatial orientation of glycosaminoglycan chains from proteoglycans. Knowledge of these changes is important to potentiate strategies aimed at restoring the morphofunctional integrity of the corneal stroma affected by limbal stem cell deficiency.

RESUMO Objetivos: Investigar a reprodutibilidade intra-la­boratorial dos fenótipos clínicos e avaliar anisotropias ópticas em córneas de coelhos com deficiência de células tronco limbais. Métodos: Lesões ao limbo foram feitas no olho direito de 23 coelhos adultos da Nova Zelândia Branco, usando um protocolo altamente agressivo, que envolveu peritomia limbal em 360 graus, ceratolimbectomia, cauterização por álcali, e remoção mecânica de epitélio remanescente. Os olhos foram clinicamente avaliados por 28 dias, inclusive por citologia de impressão corneal. As córneas que manifestaram um conjunto de alterações típicas de deficiência de células tronco limbais foram coletadas e submetidas à estudos em histopatologia e em anisotropias ópticas. Córneas saudáveis foram usadas como controles. Diferenças de caminho óptico de birrefringência relacionada à organização do colágeno estromal, e dicroísmo linear relacionado à expressão e à orientação das cadeias de glicosaminoglicanos dos proteoglicanos estromais, foram quantificados por microscopia de luz polarizada. Resultados: Um olho apresentou hipópio e foi excluído do estudo. Todos os olhos estudados (n=22) apresentaram sinais de inflamação ocular. A citologia de impressão não detectou células caliciformes na superfície corneal. Doze de 22 córneas manifestaram alterações clínicas típicas de deficiência de células tronco limbais, caracterizado por defeitos epiteliais, infiltrados inflamatórios, opacidade de moderada à severa, e neovascularização. Estudos por microscopia de luz polarizada mostraram que a deficiência de células tronco limbais aumentou a diferenças de caminho óptico corneal em 24,4% (versus controles). As cór­neas com deficiência de células tronco limbais foram menos dicroicas do que as córneas controle. Conclusões: Coelhos com deficiência de células tronco limbais, para aplicações em estudos pré-clínicos, devem ser rigorosamente selecionados para assegurar homogeneidade experimental, pois há evidências de que protocolos utilizados para indução de deficiência de células tronco limbais não estão associados com boa reprodutibilidade intra-laboratorial de fenótipos clínicos. A deficiência de células tronco limbais, como induzida aqui, alterou as propriedades ópticas anisotrópicas do estroma corneal. Tais alterações são indicativas de mudanças no empacotamento de colágeno e na orientação das cadeias de glicosaminoglicanos dos proteoglicanos. Conhecimentos nessas alterações são importantes para potencializar estratégias que visam a restabelecer a integridade morfofuncional do estromal corneal acometido pela deficiência de células tronco limbais.

Hyaluronic acid in dermatomyositis and polymyositis: relationship with disease and cutaneous lesions

Abstract: Background: There are scarce studies in the literature about hyaluronic acid in systemic autoimmune myopathies. Objectives: To analyze the serum level of hyaluronic acid in patients with dermatomyositis and polymyositis. Methods: Cross-sectional study, single-center, that evaluated hyaluronic acid in 18 dermatomyositis and 15 polymyositis (Bohan and Peter criteria), newly diagnosed, with clinical and laboratory activity, with no previous drug treatment. The patients were also age-, gender- and ethnicity-matched to 36 healthy individuals. The hyaluronic acid was analyzed by ELISA/EIA kit anti-hyaluronic acid. Results: There was a higher serum level of hyaluronic acid in patients with autoimmune myopathies, in relation to control group (P<0.05). Moreover, the serum level of this glycosaminoglycan was higher in dermatomyositis, when compared to polymyositis. Both groups were comparable with regard to demographic, clinical and laboratory data, except for the presence of skin lesions in the first group. Study limitations. The presence of hyaluronic acid in cutaneous lesions, particularly of patients with dermatomyositis, was not analyzed neither quantified. In addition, due to disease rarity and the establishment of strict inclusion and exclusion criteria, there was a small sample in the present study. Conclusions: As an example of others systemic autoimmune diseases, it is possible that the hyaluronic acid is involved in the pathogenesis of autoimmune myopathies, and particularly when associated with cutaneous lesions.

Study on glycosaminoglycan from Apostichopus Japonius on cellular immuno-regulation function of peripheral blood cells from pulmonary tuberculosis patients in vitro / 中国医师杂志

Objective To investigate the effects of glycosaminoglycans (HGAG) on the immune function of peripheral blood cells from patients with pulmonary tuberculosis.Methods Peripheral blood monouclear cells (PBMC) were isolated from peripheral blood of 40 healthy people (healthy group) and 30 tuberculosis patients (tuberculosis group) and cocultured with HGAG in vitro for 24 hours.Flow cytometry was used to detect the expression of CD45RA and CD45RO,as well as the expression of CD1a and CD83.Results The results showed that the expression of CD45RA and CD45RO in the tuberculosis group was the most significant (P ＜ 0.05) at the concentration of 50 μg/m coculturing with HGAG.The expression of CD45RA and CD45RO were most obvious in the healthy group at the concentration of 10 μg/ml and 50 μg/ml respectively (P ＜0.001).The difference of CD45RA between the two groups was no significant (P ＞0.05),while the difference of CD45RO was statistically significant (P ＜ 0.01) before co-culturing.The expression of CD45RA and CD45RO at 10 μg/ml and 50 μg/ml after co-culturing with HGAG were statistically significant (P ＜ 0.05).There was no statistical difference in CD1a and CD83 in healthy group before and after co-culturing (P ＞ 0.05),while there was statistically difference (P ＜ 0.05) before and after culturing in tuberculosis group.Before co-culturing,there was no significant difference in the expression of CD1a between the healthy group and the tuberculosis group (P ＞ 0.05),but CD83 expression was statistically different (P ＜ 0.001).After co-culturing,there were no significant differences in CD1a and CD83 expression between healthy and healthy groups (P ＞ 0.05).Conclusions HGAG can down-regulate the expression of CD45RA and up-regulate the expression of CD45RO in a certain concentration range,and promote the maturation of dendritic cells (DC) in tuberculosis patients and regulate the cellular immunity of patients with pulmonary tuberculosis in vitro.

Biomechanical effect of hyaluronidase on rabbit auricular cartilage / 中华医学美学美容杂志

Objective To evaluate the biomechanical effectiveness of hyaluronidase (HAase) on rabbit auricular cartilage in the early stage.Methods HAase in 3 different concentrations (75 U/ml,150 U/ml,300 U/ml) were injected subcutaneously to rabbit auricle in 3 groups,while normal saline were injected as control.For the comparison of biomechanism properties among different groups and different time,cartilages were harvested at 3rd and 7th day after injection,followed by stress-relaxation assay.Results Auricular cartilage displayed different levels in control group compared with other groups in elastic modulus (P＜0.05) and maxmum stress (P＜0.05) in 3th day as well as 7th day.While both in the same concentration group,there were also differences between the 3th and 7th day (P＜0.05).Conclusions HAase injection can cause changes in biomechanical properties of auricular cartilage.And 7 days would not be enough for the tissue recovery biomechanically.

Endovenous laser ablation combined with foam sclerotherapy and mucopolysaccharide polysulfate cream for the treatment of stasis dermatitis: a clinical trial / 中华皮肤科杂志

Objective To evaluate the therapeutic effect of endovenous laser ablation (ELA) combined with foam sclerotherapy (FS) and mucopolysaccharide polysulfate (MP)cream on stasis dermatitis.Methods From December 2015 to May 2017,52 patients with 60 lesional limbs were enrolled from Department of Dermatology of Sichuan Provincial People's Hospital.The 60 lesional limbs were randomly and equally divided into 3 groups by a random number table and remainder grouping:combination group,MP group and control group.All the 3 groups were firstly treated with ELA in the main great saphenous vein.Then,the combination group was treated with FS followed by topical MP cream for 4 weeks.After the laser therapy,the MP group was treated with topical MP cream for 4 weeks,and the control group was treated with topical mometasone furoate cream alone for 4 weeks.The eczema area and severity index (EASI) and visual analogue scale (VAS) for itching scores in the above 3 group were recorded before treatment and 4 weeks after treatment.Statistical analysis was done by paired t-test for comparisons before and after treatment,one-way analysis of variance (ANOVA) for intergroup comparison,and least significant difference (LSD)-t test for multiple comparisons.Results No significant difference was observed before treatment among the combination group,MP group and control group in the EASI (9.64 ± 4.58,9.94 ± 4.18,9.50 ± 4.41 respectively,F =0.052,P ＞ 0.05) or VAS scores (7.25 ± 1.29,7.50 ± 1.19,7.45 ± 1.32 respectively,F =0.218,P ＞ 0.05).After 4-week treatment,the combination group,MP group and control group all showed significantly decreased EASI (3.54 ± 1.57,5.86 ± 2.39,7.04 ± 2.75 respectively) and VAS scores (2.35 ± 0.67,3.85 ± 0.67,4.65 ± 1.23 respectively) compared with those before treatment (t =4.30-18.80,all P ＜ 0.05).After 4-week treatment,the EASI score was significantly lower in the combination group than in the MP group and control group (both P ＜ 0.05),while there was no significant difference between the MP group and control group (P ＞ 0.05).Additionally,the VAS score was significantly lower in the combination group than in the MP group and control group (both P ＜ 0.05),as well as in the MP group than in the control group (P ＜ 0.05).Conclusions ELA combined with FS and topical MP cream shows better short-term efficacy for the treatment of stasis dermatitis compared with ELA combined with topical MP cream or mometasone furoate cream.The combination with topical MP cream is superior to that with topical mometasone furoate cream in improving itching.The long-term efficacy needs to be observed further.

The effects of varus degree on the early metabolic changes of the lateral compartment cartilage in knees with medial unicompartmental osteoarthritis / 中华骨科杂志

Objective To find the effects of varus degree on the early metabolic changes of the lateral compartment cartilage in knees with medial unicompartmental osteoarthritis by detecting glycosaminoglycan (GAG) in varus knees.Methods From June 2016 to December 2017,twenty middle-aged volunteers without osteoarthritis or coronal deformities were recruited as the control group.Sixty patients diagnosed as medial unicompartmental osteoarthritis were recruited as the osteoarthritis group.The patients were further divided into four groups according to the degrees of varus angle,namely 2°-5° varus group,5°-10° varus group,10°-15° varus group and ＞15° varus group with 15 patients in each group.Delayed gadolinium-enhanced magnetic resonance imaging of cartilage (dGEMRIC) was performed in all participants.The dGEMRIC indices (T1Gd) representing GAG content were calculated in the weight-bearing femoral cartilage (wbFC),the posterior non-weight-bearing femoral cartilage (pFC),the lateral femoral cartilage (FC,wbFC+pFC) and the tibial cartilage (TC) in the lateral compartment by using Matlab 7.1 and MRIMapper software.Results T1Gd of wbFC,pFC,FC and TC were 400.3±51.5 ms,393.6±57.9 ms,397.5±52.3 ms and 448.6±62.5 ms in the control group,391.8±41.5 ms,407.2±43.8 ms,400.1±37.8 ms and 461.3±41.6 ms in 2°-5° varus group and 386.9±57.1 ms,401.3±73.5 ms,397.7±59.6 ms and 438.9±42.8 ms in 5°-10° varus group.There was no significant difference among the above three groups in T1Gd in any of the analyzed cartilage regions (P＞0.05).In 10°-15° varus group,T1Gd of wbFC,pFC,FC and TC were 380.1±45.5 ms,385.5±76.6 ms,384.0±53.5 ms and 400.2±43.8 ms,respectively.Although T1Gd of wbFC,pFC and FC in 10°-15° varus group were similar with that in the control group,2°-5° varus group and 5°-10° varus (P＞0.05),T1Gd of TC in 10°-15° varus group decreased significantly (P＜0.05).In addition,T1Gd of wbFC,pFC,FC and TC in ＞15° varus groupwere 327.7±54.3 ms,340.1±33.0 ms,334.9±36.0 ms and 363.6±48.6 ms,respectively.T1Gd of all regions of interest in ＞15° varus group were significantly lower than that informer four groups (P＜0.05).Conclusion In medial unicompartmental knee osteoarthritis,there is a relationship between varus degree and GAG content of the lateral compartment cartilage.If varus angle ≤10°,the GAG content of the lateral compartment cartilage was similar with the similar aged subjects without osteoarthritis.If varus angle ＞ 10°,GAG content of the lateral compartment decreases significantly.

Nondestructive Assessment of Glycosaminoglycans in Engineered Cartilages Using Hexabrix-Enhanced Micro-Computed Tomography

It is very useful to evaluate the content and 3D distribution of extracellular matrix non-destructively in tissue engineering. This study evaluated the feasibility of using micro-computed tomography (µCT) with Hexabrix to measure quantitatively sulfated glycosaminoglycans (GAGs) of engineered cartilage. Rabbit chondrocytes at passage 2 were used to produce artificial cartilages in polyglycolic acid scaffolds in vitro. Engineered cartilages were incubated with Hexabrix 320 for 20 min and analyzed via µCT scanning. The number of voxels in the 2D and 3D scanning images were counted to estimate the amount of sulfated GAGs. The optimal threshold value for quantification was determined by regression analysis. The 2D µCT images of an engineered cartilage showed positive correlation with the histological image of Safranin-O staining. Quantitative data obtained with the 3D µCT images of 14 engineered cartilages showed strong correlation with sulfated GAGs contents obtained by biochemical analysis (R² = 0.883, p < 0.001). Repeated exposure of engineered cartilages to Hexabrix 320 and µCT scanning did not significantly affect cell viability, total DNA content, or the total content of sulfated GAGs. We conclude that µCT imaging using Hexabrix 320 provides high spatial resolution and sensitivity to assess the content and 3D distribution of sulfated GAGs in engineered cartilages. It is expected to be a valuable tool to evaluate the quality of engineered cartilage for commercial development in the future.