Metformin and resveratrol suppress type 2 diabetes mellitus-induced articular cartilage damage in rats associated with the inhibition of inflammation and augmentation of proteoglycans / La metformina y el resveratrol suprimen el daño del cartílago articular inducido por la diabetes mellitus tipo 2 en ratas asociado con la inhibición de la inflamación y el aumento de proteoglicanos

SUMMARY: Induction of osteoarthritis (OA) following diabetes is characterized by a sever inflammation of the joints that can lead to disability. The cartilage content of proteoglycans can substantially be reduced, following the induction of diabetes mellitus associated with inflammation as well as knee joint injury, and the antidiabetic drug metformin combined with the anti-inflammatory agent resveratrol can prevent these deleterious effects. Therefore, insulin-independent diabetes, type 2 diabetes mellitus (T2DM) was induced in Albino rats by streptozotocin (STZ) injection (50 mg/kg) after being fed on a high carbohydrate and fat diets for 2 weeks. The protective group of rats which also received a single injection of STZ was treated daily with metformin (Met; 200 mg/kg) and resveratrol (Res; 30 mg/kg) for 12 weeks. Harvested knee joint tissues were prepared for basic histology stain and for proteoglycans staining using light microscopy. Histology images showed in diabetic rats (T2DM) OA development as demonstrated by profound injury to the knee joint and severe decrease of articular cartilage proteoglycans content, which were substantialy protected by Met+Res. Met+Res also significantly (p< 0.0001) decreased diabetes induced glycemia, dyslipidemia, and the inflammatory biomarkers, tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), and high sensitivity C-reactive protein (hs-CRP). In addition, there was a significant correlation between OA and glycemia, dyslipidemia, and inflammation. Collectively, we demonstrate an association between knee joint damage and biomarkers of glycemia, dyslipidemia, and inflammation in diabetes-induced OA, with metformin plus resveratrol providing protective effects.

RESUMEN: La inducción de osteoartritis (OA) después de la diabetes se caracteriza por una inflamación severa de las articulaciones que puede conducir a la discapacidad. El contenido de cartílago de proteoglicanos se puede reducir sustancialmente, luego de la inducción de diabetes mellitus asociada con inflamación y lesión en la articulación de la rodilla sin embargo, el fármaco antidiabético metformina combinado con el agente antiinflamatorio resveratrol puede prevenir estos efectos nocivos. Por lo tanto, se indujo diabetes insulino dependiente, diabetes mellitus tipo 2 (T2DM) en ratas albinas mediante inyección de estreptozotocina (STZ) (50 mg/kg) después de haber sido alimentadas con dietas ricas en carbohidratos y grasas durante 2 semanas. El grupo protector de ratas que también recibió una inyección única de STZ fue tratado diariamente con metformina (Met; 200 mg/kg) y resveratrol (Res; 30 mg/kg) durante 12 semanas. Tejidos de la articulación de la rodilla fueon retirados y teñidos con histología básica y tinción de proteoglicanos usando microscopía óptica. Las imágenes histológicas en ratas diabéticas mostraban (T2DM) desarrollo de OA visualizadas por una lesión profunda en la articulación de la rodilla y una disminución severa del contenido de proteoglicanos del cartílago articular, los cuales estaban sustancialmente protegidos por Met+Res. Met+Res. También disminuyó significativamente (p< 0,0001) la glucemia inducida por la diabetes, la dislipidemia y los biomarcadores inflamatorios, el factor de necrosis tumoral alfa (TNF-α), la interleucina-6 (IL-6) y la proteína C reactiva de alta sensibilidad (PCR-hs). Además, hubo una correlación significativa entre la OA y la glucemia, la dislipidemia y la inflamación. En conjunto, demostramos una asociación entre el daño de la articulación de la rodilla y los biomarcadores de glucemia, dislipidemia e inflamación en la OA inducida por diabetes, con metformina más resveratrol que brindan efectos protectores.

Effects of small leucine-rich proteoglycans on corneal transparency / 中华实验眼科杂志

Small leucine-rich proteoglycans (SLRPs) are necessary structural ingredients of the cornea, which are vital for the establishment and maintenance of corneal transparency.SLRPs are mainly located in the corneal stroma and can be divided into class Ⅰ, class Ⅱ, and class Ⅲ.The compensatory and cooperative interactions among SLRPs regulate the formation and assembly of stromal collagen fibrils, thereby maintaining the highly ordered arrangement of collagen fibrils, and establishing corneal transparency.Decorin and lumican are the main functional components of class Ⅰ and class Ⅱ SLRPs, respectively, and changes in their expression or abnormities in the structure of their core proteins affect the natural content and arrangement of other stromal extracellular matrix components, ultimately resulting in abnormal fibril formation, assembly, and arrangement, causing corneal opacity.SLRPs can regulate corneal wound healing and stromal matrix remodeling via binding to fibrotic molecules and their receptors, which provides bases for corneal diseases therapy and study of molecular mechanisms of corneal transparency.The bioactivities and the role of SLRPs in corneal transparency were reviewed in this article.

Heparan sulfate proteoglycan triggers focal adhesion kinase signaling during Trypanosoma cruzi invasion

BACKGROUND Trypanosoma cruzi, the etiologic agent of Chagas disease, is capable of triggering different signaling pathways that modulate its internalisation in mammalian cells. Focal adhesion kinase (FAK), a non-receptor tyrosine kinase protein, has been demonstrated as a mechanism of T. cruzi invasion in cardiomyocytes. Since the involved cell surface receptors are not yet known, we evaluated whether heparan sulfate proteoglycans (HSPG), a molecule involved in T. cruzi recognition and in the regulation of multiple signaling pathways, are able to trigger the FAK signaling pathway during T. cruzi invasion. METHODS To investigate the role of HSPG in the regulation of the FAK signaling pathway during trypomastigote entry, we performed heparan sulfate (HS) depletion from the cardiomyocyte surface by treatment with heparinase I or p-nitrophenyl-β-D-xylopyranoside (p-n-xyloside), which abolishes glycosaminoglycan (GAG) attachment to the proteoglycan core protein. Wild-type (CHO-k1) and GAG-deficient Chinese hamster ovary cells (CHO-745) were also used as an approach to evaluate the participation of the HSPG-FAK signaling pathway. FAK activation (FAK Tyr397) and spatial distribution were analysed by immunoblotting and indirect immunofluorescence, respectively. FINDINGS HS depletion from the cardiomyocyte surface inhibited FAK activation by T. cruzi. Cardiomyocyte treatment with heparinase I or p-n-xyloside resulted in 34% and 28% FAK phosphorylation level decreases, respectively. The experiments with the CHO cells corroborated the role of HSPG as a FAK activation mediator. T. cruzi infection did not stimulate FAK phosphorylation in CHO-745 cells, leading to a 36% reduction in parasite invasion. FAK inhibition due to the PF573228 treatment also impaired T. cruzi entry in CHO-k1 cells. MAIN CONCLUSION Jointly, our data demonstrate that HSPG is a key molecule in the FAK signaling pathway activation, regulating T. cruzi entry.

Is the expression of the components of the carotid matrix of rats influenced by estrogen, progestin and tibolone? / A expressão dos componentes da matriz carotídea de ratos é influenciada pelo estrogênio, progestagênio e tibolona?

Abstract Objective To analyze the effects of estrogen alone or in combination with progestogens and tibolone (TIB) on the expression of the extracellular matrix metalloproteinases 2 and 9 (MMP-2 and MMP-9), of perlecan, and of heparanase (HPSE) of the vascular walls of the carotid arteries. Methods A total of 30 250-day-old ovariectomized Wistar rats were orally treated for 5 weeks with: a) 1 mg/kg of estradiol benzoate (EB); b) EB + 0.2 mg/kg of medroxyprogesterone acetate (MPA); c) EB + 0.2mg/kg of norethisterone acetate (NETA); d) EB + 2 mg/kg of dydrogesterone (DI); e) 1 mg/kg of TIB; f) placebo (CTR). Following treatment, the expression of mRNA for MMP-2, MMP-9, and HPSE was analyzed by realtime polymerase chain-reaction (PCR), and the expression of MMP-2, of MMP-9, of tissue inhibitor of metalloproteinase 2 (TIMP-2), and of perlecan was quantified by immunohistochemistry in the carotid arteries. Results The groups showed significant differences on mRNA HPSE expression (p = 0.048), which was higher in the EB, EB + MPA, and TIB groups. There was no statistically significant difference in mRNA MMP-2 or MMP-9 expression. The immunohistochemical expression of MMP-2, of TIMP-2, of MMP-9, of HPSE, and of perlecan showed no differences between groups. Conclusion Estradiol alone or associated with MPA and TIB treatment can increase mRNA HSPE expression of the walls of the carotid arteries in ovariectomized rats.

Resumo Objetivo Analisar os efeitos do estrogênio isolado ou em combinação com progestogênios e tibolona (TIB) na expressão das metaloproteinases 2 e 9 da matriz extracelular (MMP-2 e MMP-9), da perlecan e da heparanase (HPSE) das paredes vasculares das artérias carótidas. Métodos Trinta ratas Wistar ovariectomizadas com 250 dias de idade foram tratadas oralmente por 5 semanas com: a) 1 mg/kg de benzoato de estradiol (EB); b) EB + 0,2 mg/kg de acetato de medroxiprogesterona (MPA); c) EB + 0,2mg/kg de acetato de noretisterona (NETA); d) EB + 2 mg/kg de didrogesterona (DI); e) 1 mg/kg de TIB; f) placebo (CTR). Após o tratamento, a expressão de mRNA para MMP-2, MMP- 9, e HPSE foi analisada por reação em cadeia da polimerase (RCP) em tempo real, e a expressão de MMP-2, MMP-9, inibidor tecidual de metaloproteinase 2 (TIMP-2), e de perlecan foi quantificado por imunohistoquímica em artérias carótidas. Resultados Os grupos apresentaram diferenças significativas na expressão do mRNA HPSE (p = 0,048), sendo maiores nos grupos EB, EB + MPA e TIB. Não houve diferença estatisticamente significativa nas expressões de mRNA MMP-2 ou MMP-9. A expressão imunohistoquímica de MMP-2, TIMP-2, MMP-9, HPSE e perlecan não mostrou diferenças entre os grupos. Conclusão O estradiol isolado ou associado ao tratamento com MPA e TIB pode aumentar a expressão de mRNA HSPE nas paredes das artérias carótidas em ratas ovariectomizadas.

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.

Differential gene expression analysis of human cumulus cells / 대한생식의학회지

OBJECTIVE: This study was performed to explore the possibility that each oocyte and its surrounding cumulus cells might have different genetic expression patterns that could affect human reproduction. METHODS: Differential gene expression analysis was performed for 10 clusters of cumulus cells obtained from 10 cumulus-oocyte complexes from 10 patients. Same procedures related to oocyte maturation, microinjection, and microarray analyses were performed for each group of cumulus cells. Two differential gene expression analyses were performed: one for the outcome of clinical pregnancy and one for the outcome of live birth. RESULTS: Significant genes resulting from these analyses were selected and the top 20 affected pathways in each group were analyzed. Circadian entrainment is determined to be the most affected pathway for clinical pregnancy, and proteoglycans in cancer pathway is the most affected pathway for live birth. Circadian entrainment is also amongst the 12 pathways that are found to be in top 20 affected pathways for both outcomes, and has both lowest p-value and highest number of times found count. CONCLUSION: Although further confirmatory studies are necessary, findings of this study suggest that these pathways, especially circadian entrainment in cumulus cells, may be essential for embryo development and pregnancy.

Molecular characterization of CHST11 and its potential role in nacre formation in pearl oyster Pinctada fucata martensii

Background: C4ST-1 catalyzes the transfer of sulfate groups in the sulfonation of chondroitin during chondroitin sulfate synthesis. Chondroitin sulfate consists of numerous copies of negatively charged sulfonic acid groups that participate in the nucleation process of biomineralization. In the present study, we obtained two CHST11 genes (PmCHST11a and PmCHST11b) which encoded the C4ST-1 and explored the functions of these genes in the synthesis of chondroitin sulfate and in the formation of the nacreous layer of shells. Results: Both PmCHST11a and PmCHST11b had a sulfotransferase-2 domain, a signal peptide and a transmembrane domain. These properties indicated that these genes localize in the Golgi apparatus. Real-time PCR revealed that both PmCHST11a and PmCHST11b were highly expressed in the central zone of the mantle tissue. Inhibiting PmCHST11a and PmCHST11b via RNA interference significantly decreased the expression levels of these genes in the central zone of the mantle tissue and the concentration of chondroitin sulfate in extrapallial fluid. Moreover, shell nacre crystallized irregularly with a rough surface after RNA interference. Conclusions: This study indicated that PmCHST11a and PmCHST11b are involved in the nacre formation of Pinctada fucata martensii through participating in the synthesis of chondroitin sulfate.

SRGN enhances the breast cancer cells metastasis by inducing Epithelial-mesenchymal transition / 中国医师杂志

Objective To investigate the mechanism of glycoprotein serglycin (SRGN) promoting metastasis of breast cancer cells and the possible mechanism of SRGN expression.Methods Real time quantitative polymerase chain reaction (PCR) and bioinformation retrieval were used to detect the expression of SRGN in lymph node metastasis and non-metastasis breast cancer.MDA-MB-231 shRNA and MCF-7-SRGN of breast cancer stable cell line were established by lentivirus shRNA interferencc and overexpression.Transwell assay was used to test the effect of SRGN on invasion and metastasis of breast cancer cell line in vitro.Western blot assay was used to detect the changes of epithelial-mesenchymal (EMT) related markers.The possible regulatory mechanism of SRGN expression was detected by Western blot assay.Results SRGN expression was significantly increased in lymph node metastasis of breast cancer in clinical specimens.SRGN interference inhibited the invasion and metastasis of tumor cells.SRGN promoted breast cancer cells EMT.Transforming growth factor β1 (TGFβ1) promoted the expression of beta SRGN transcription.Conclusions SRGN can induce the change of EMT in breast cancer cells and promote the invasion and metastasis of breast cancer cells.

Effect of dentin proteoglycans on the stability of resin-dentin bonds against artificial saliva storage / 中华口腔医学杂志

Objective@#To investigate the potential effect of proteoglycans (PG) and glycosaminoglycans (GAG) on the stability of resin-dentin bonds against artificial saliva storage.@*Methods@#Seventy-two extracted molars were used to obtain standard dentin bonding surface, and the specimens were etched for 15 s with 37% phosphoric acid and divided into three groups using a table of random number. Then the three groups undergone different incubating procedures as follow: specimens in chondroitinase ABC (C-ABC) group were incubated with C-ABC, specimens in trypsin (TRY) group were incubated with trypsin, and specimens in the control group were incubated with deionized water. All specimens were incubated at 37 ℃ for 48 h in the oscillators. Then specimens in each group were randomly assigned into three subgroups (n=8) as follows: immediate control subgroup, aging subgroups with artificial saliva storage for 6 months and 12 months. Microtensile bond strength (μTBS), fracture mode, bonding interface morphology and nanoleakage were evaluated.@*Results@#Immediately and with artificial saliva storage for 6 months and 12 mouths, the μTBS of TRY group ([49.04±3.57], [37.01±3.21] and [35.27±3.56] MPa) were significantly higher than those in the control group ([40.71±3.32], [28.87±2.34] and [24.20±2.07] MPa) (P<0.05). The immediate μTBS of C-ABC group ([32.94±2.45] MPa) was significantly lower than that of the control group (P<0.05). While with artificial saliva storage for 6 months and 12 mouths, the μTBS of C-ABC group ([26.46±2.45] and [22.50±2.58] MPa) were no differences with those of the control group (P>0.05). The ratio of cohesive fracture increased with the extension of aging time. Some narrow gaps were found in hybrid layer of the control group with artificial saliva storage for 6 months and 12 mouths.@*Conclusions@#Removal of PG increased the μTBS and durable bonds to dentin, while removal of GAG decreased the μTBS, however, it can be of help to create more durable bonds to dentin.

The relationship between serum aggrecan catabolic fragments and joint Sharp assessment in patients with rheumatoid arthritis / 中华风湿病学杂志

Objective To investigate the relationship between abnormal metabolism of aggrecan and joint destruction in patients with rheumatoid arthritis (RA).Methods 140 RA patients with duration less than 24 months were enrolled into this study.The study also included 100 normal controls and 95 patients with other rheumatic diseases.Three monoclonal antibodies (5D4,7D4 and BC-3) of aggrecan were used to detected aggrecan catabolic fragments in serum of RA patients and the other two groups of controls by enzyme linked immunosorbent assay (ELISA),and the correlation of aggrecan catabolic fragments with joint damage were analyzed.Sharp evaluation of hand joints in RA patients were performed at baseline and after one year follow-up.Calculating the area under the receiver operating characteristic curve (ROC) was used to evaluate the sensitivity and specificity of aggrecan catabolic fragments detected in serum of RA patients.Results Both levels of 5D4 fragment and BC-3 fragment of RA group were higher than those of normal control [5D4 of RA:(5.8±2.1) ng/μl,normal control:(2.2±1.3) ng/μl;BC-3 of RA:(11.1±3.4) ng/μl,normal control:(5.0±2.1) ng/μl,F=38.65,24.07,P＜0.001).There was no difference in 7D4 fragment among three groups (F=0.589,P=0.478).Both two fragment levels of RA patients with anti-CCP positive were greater than those patients with anti-CCP negative [5D4:(5.6±1.3) ng/μl vs (4.4±1.1) ng/μl,F=21.23,P＜0.01;BC-3:(12.2±3.9) ng/μl vs (9.3±2.8) ng/μ1,F=27.14,P＜0.01].Linear Regression showed that serum fragments detected by 5D4 and BC-3,and anti-CCP positive were risk factors for Sharp deterioration after one year follow-up.The sensitivity and specificity of combined detection of two aggrecan fragments in serum of RA patients for the prediction of joint Sharp were 56.5％ and 84.2％ respectively.Positive predictive value and negative predictive value are 74.3％ and 70.6％.respectively.Application of areas of ROC to identify the best evaluation of Sharp was 0.798.Conclusion There is positive correlation between aggrecan catabolic fragments in serum and joint Sharp evaluation of RA patients.Detection of aggrecan catabolic fragments in RA patients may predict early joint destruction.

Glicosaminoglicanos e proteoglicanos na fáscia palmar de pacientes com dupuytren / Glycosaminoglycans and proteoglycans in palmar fascia of patients with dupuytren

Objetivo: Avaliar e comparar o comportamento dos glicosaminoglicanos(GAGs) na Doença de Dupuytren (DD). Métodos: Trata-se deum estudo experimental com 23 pacientes diagnosticados com DD.Tecidos coletados através de fasciectomia com incisão tipo Brunnerou McCash, foram avaliados por eletroforese para identificação dosGAGs. A quantificação foi realizada por imunofluorescência e dosagemdas proteínas para os diferentes tipos de glicosaminoglicanos.Os resultados coletados foram expressos em porcentagem e avaliadosestatisticamente. Resultados: Foi observado, na corrida eletroforética,aumento significativo dos GAGs em relação ao controle(p<0,05). Na imunofluorescência houve redução (23 vezes) do ácidohialurônico quando comparado ao controle (p<0,0001). Conclusão:Ficou evidenciado pelos resultados o aumento dos GAGs sulfatadosna doença de Dupuytren, principalmente do dermatam sulfato, egrande diminuição do ácido hialurônico na aponeurose palmar dosmesmos pacientes. Nível de Evidência III, Estudo Caso Controle.

Objective: To evaluate and compare the behavior of glycosaminoglycans(GAGs) in Dupuytren disease (DD). Methods: Thisis an experimental study with 23 patients diagnosed with DD.Tissue collected through fasciectomy with incision type Brunneror McCash were evaluated by electrophoresis for identificationof GAGs. The quantification was carried out by immunofluorescenceand dosage of proteins for different types of glycosaminoglycans.The results were expressed in percentage and statisticallyevaluated. Results: A significant increase was observed througheletrophoresis in GAGs, as compared to the control (p<0.05). Immunofluorescenceof hyaluronic acid was reduced (23 times) whencompared to the control (p<0.0001). Conclusion: An increase ofsulfated GAGs in Dupuytren’s disease, mainly dermatan sulfate,was evident from our results, as well as a pronounced decrease ofhyaluronic acid in the palmar aponeurosis from the same patients.Level of Evidence III, Case-Control Study.

Isolation and Quantification of Glycosaminoglycans from Human Hair Shaft

BACKGROUND: There is evidence that glycosaminoglycans (GAGs) are present in the hair shaft within the follicle but there are no studies regarding GAGs isolation and measurement in the human hair shaft over the scalp surface, it means, in the free hair shaft. OBJECTIVE: The purpose of our research was to isolate and measure the total GAGs from human free hair shaft. METHODS: Seventy-five healthy individuals participated in the study, 58 adults, men and women over the age of 50 and 17 children (aged 4~9). GAGs in hair samples, received from the parietal and the occipital areas, were isolated with 4 M guanidine HCl and measured by the uronic acid-carbazole reaction assay. RESULTS: GAGs concentration was significantly higher in the occipital area than in the parietal area, in all study groups. GAG levels from both areas were significantly higher in children than in adults. GAG levels were not associated with gender, hair color or type. CONCLUSION: We report the presence of GAGs in the human free hair shaft and the correlation of hair GAG levels with the scalp area and participants' age.

Glycomics expression analysis of sulfated glycosaminoglycans of human colorectal cancer tissues and non-neoplastic mucosa by electrospray ionization mass spectrometry / Análise da expressão de glicosaminoglicanos sulfatados no tecido humano neoplásico colorretal e na mucosa não neoplásica por espectrometria de massa por ionização por electrospray

ABSTRACT Objective To determine the presence of glycosaminoglycans in the extracellular matrix of connective tissue from neoplastic and non-neoplastic colorectal tissues, since it has a central role in tumor development and progression. Methods Tissue samples from neoplastic and non-neoplastic colorectal tissues were obtained from 64 operated patients who had colorectal carcinoma with no distant metastases. Expressions of heparan sulphate, chondroitin sulphate, dermatan sulphate and their fragments were analyzed by electrospray ionization mass spectrometry, with the technique for extraction and quantification of glycosaminoglycans after proteolysis and electrophoresis. The statistical analysis included mean, standard deviation, and Student’st test. Results The glycosaminoglycans extracted from colorectal tissue showed three electrophoretic bands in agarose gel. Electrospray ionization mass spectrometry showed characteristic disaccharide fragments from glycosaminoglycans, indicating their structural characterization in the tissues analyzed. Some peaks in the electrospray ionization mass spectrometry were not characterized as fragments of sugars, indicating the presence of fragments of the protein structure of proteoglycans generated during the glycosaminoglycan purification. The average amount of chondroitin and dermatan increased in the neoplastic tissue compared to normal tissue (p=0.01). On the other hand, the average amount of heparan decreased in the neoplastic tissue compared to normal tissue (p= 0.03). Conclusion The method allowed the determination of the glycosaminoglycans structural profile in colorectal tissue from neoplastic and non-neoplastic colorectal tissue. Neoplastic tissues showed greater amounts of chondroitin sulphate and dermatan sulphate compared to non-neoplastic tissues, while heparan sulphate was decreased in neoplastic tissues.

RESUMO Objetivo Determinar a presença de glicosaminoglicanos na matriz extracelular do tecido conjuntivo colorretal neoplásico e não neoplásico, tendo em vista seu papel central no desenvolvimento e na progressão dos tumores. Métodos Amostras de tecidos colorretais neoplásicos e não neoplásicos foram obtidas de 64 pacientes operados com carcinoma colorretal sem metástases a distância. As expressões de heparan sulfato, sulfato de condroitina e sulfato de dermatan e seus fragmentos foram analisadas por espectrometria de massa por ionização por electrospray, com técnica de extração e quantificação de glicosaminoglicanos após proteólise e eletroforese. Para análise estatística, utilizaram-se média, desvio padrão e teste t de Student. Resultados Em gel de agarose, os glicosaminoglicanos extraídos de tecido colorretal mostraram três bandas eletroforéticas. A espectrometria de massa por ionização por electrospray mostrou fragmentos de dissacarídeos característicos de glicosaminoglicanos e indicou sua característica estrutural. Alguns picos na espectrometria de massa por ionização por electrospray não foram caracterizados como fragmentos de açúcares, sugerindo a presença de fragmentos de proteínas estruturais dos proteoglicanos, formadas durante a purificação dos glicosaminoglicanos. A quantidade média de condroitina e dermatan aumentou no tecido neoplástico em relação ao tecido normal (p=0,01). Por outro lado, a quantidade média de heparan foi menor no tecido neoplásico em relação ao tecido normal (p=0,03). Conclusão O método empregado permitiu determinar o perfil estrutural dos glicosaminoglicanos nas amostras. Tecidos neoplásicos apresentaram maiores quantidades de sulfato de condroitina e sulfato de dermatan em comparação com os não neoplásicos, enquanto o sulfato de heparan foi encontrado em menores quantidades nos tecidos neoplásicos.

Proteoglycan-induced arthritis model in mice: Research progress / 第二军医大学学报

Proteoglycan-induced arthritis (PGIA) is a unique systemic autoimmune disease model of spondyloarthritis (SpA) induced by intraperitoneal immunization of either BALB/c or certain C3H colonies with cartilage proteoglycan. This model is currently the only systemic autoimmune mice model with axial skeleton manifestation, which has become a focus of study in SpA modeling. Here in this review we summarize the methods of animal model building, phenotype identification, immunological characteristics and pathways involved in bone remodeling of PGIA based on previous studies, hoping to provide references for study on SpA animal model.

Evaluation of influence of proteoglycans on hydration of articular cartilage with the use of ultrasound / 解放军医学杂志

Objective To monitor the changes in hydration behaviour of articular cartilage induced by degradation of proteoglycans, and to explore the effect of proteoglycans on hydration behaviour of articular cartilage by using high-frequency ultrasound. Methods Twelve porcine patellae with smooth cartilage surface were prepared and equally divided into two groups: normal group without any enzyme treatment, and trypsin group they were treated with 0.25% trypsin for 8h to digest proteoglycan in the cartilage. The hydration behaviour of the cartilage tissue was scanned by high-frequency ultrasound system with a central frequency of 25MHz. Parameters including cartilage hydration strain and cartilage thickness were measured. The histopathological changes in the articular cartilage were observed under a light microscope. Results It took approximately 20min to reach equilibrium during the hydration process in the normal cartilages, while proteoglycan-degraded cartilage took only about 5min to achieve equilibrium. The equilibrium strain of normal cartilage was 3.5%±0.5%. The degradation of proteoglycans induced a significant decrease in equilibrium strain (1.8%±0.2%, P0.05). Conclusion Proteoglycans play an important role in hydration behaviour of articular cartilage. The degradation of proteoglycans could induce degeneration of cartilage structure and decrease in hydration behaviour after dehydration.

Effect of chondroitin sulfate proteoglycans on the repair of nerve system injury / 中国生化药物杂志

Chondroitin sulfate proteoglycans ( CSPGs) is one kind of proteins that covalently bind with chondroitin sulfate.CSPGs play important roles in the growth and development of the central nervous system and the pathological reaction of nervous injury.This article reviews the functional and mechanism studies of CSPGs in the repair of nerve system injury.

From DCPD to NTCP: The long journey towards identifying a functional hepatitis B virus receptor

Hepatitis B virus (HBV) is the prototype of hepatotropic DNA viruses (hepadnaviruses) infecting a wide range of human and non-human hosts. Previous studies with duck hepatitis B virus (DHBV) identified duck carboxypeptidase D (dCPD) as a host specific binding partner for full-length large envelope protein, and p120 as a binding partner for several truncated versions of the large envelope protein. p120 is the P protein of duck glycine decarboxylase (dGLDC) with restricted expression in DHBV infectible tissues. Several lines of evidence suggest the importance of dCPD, and especially p120, in productive DHBV infection, although neither dCPD nor p120 cDNA could confer susceptibility to DHBV infection in any cell line. Recently, sodium taurocholate cotransporting polypeptide (NTCP) has been identified as a binding partner for the N-terminus of HBV large envelope protein. Importantly, knock down and reconstitution experiments unequivocally demonstrated that NTCP is both necessary and sufficient for in vitro infection by HBV and hepatitis delta virus (HDV), an RNA virus using HBV envelope proteins for its transmission. What remains unclear is whether NTCP is the major HBV receptor in vivo. The fact that some HBV patients are homozygous with an NTCP mutation known to abolish its receptor function suggests the existence of NTCP-independent pathways of HBV entry. Also, NTCP very likely mediates just one step of the HBV entry process, with additional co-factors for productive HBV infection still to be discovered. NTCP offers a novel therapeutic target for the control of chronic HBV infection.

Expression of Hyaluronidase-4 in a Rat Spinal Cord Hemisection Model

STUDY DESIGN: Examination of hyaluronidase-4 (Hyal-4) expression in a rat spinal cord hemisection model. PURPOSE: To determine the status of Hyal-4 expression after hemisection of the spinal cord, and the relationship between its expression and that of chondroitin sulfate proteoglycans (CSPGs). OVERVIEW OF LITERATURE: CSPGs are expressed at the site of spinal cord injury and inhibit axon regeneration. Administration of exogenous chrondroitinase ABC (ChABC), derived from bacteria, digested CSPGs and promoted axonal regrowth. Using a rat hemisection model, we have demonstrated peak CSPGs levels at by 3 weeks after injury but then decreased spontaneously. Could there be an endogenous enzyme similar to ChABC in the spinal cord? It has been suggested that Hyal-4 is involved in CSPG degradation. METHODS: A rat hemisection model was prepared and spinal cord frozen sections were prepared at 4 days and 1, 2, 3, 4, 5, and 6 weeks post-cordotomy and stained for CSPGs and Hyal-4 and subjected to Western blotting. RESULTS: CSPGs appeared at the injury site at 4 days after hemisection, reached a peak after 3 weeks, and then decreased. Hyal-4 was observed around the injury site from 4 days after cordotomy and increased until after 5-6 weeks. Double staining showed Hyal-4 around CSPGs. Western blotting identified a band corresponding to Hyal-4 from 4 days after hemisection. CONCLUSIONS: Hyal-4 was expressed in a rat hemisection model in areas surrounding CSPGs, and as its peak was delayed compared with that of CSPGs. These results suggest the involvement of Hyal-4 in the digestion of CSPGs.

MicroRNA181b/c inhibits the metastasis of breast cancer by targeting glycoprotein serglycin / 中国医师杂志

Objective To investigate the relationship of glycoprotein serglycin (SRGN) expression with invasion and metastasis of breast cancer cells,and the role of microRNA in the regulation of SRGN expression.Methods Real-time quantitative polymer ase chain reaction (PCR) and Western blot were used to detect the differences in SRGN expression between higher metastasis Michigan cancer foundation-7 (MCF-7)/5-Fu breast cancer cell lines and weaker metastasis MCF-7 cell line.The siRNA interference experiment and in vitro Transwell experiment were used to detect effect of SRGN on the ability of invasion and metastasis of breast cancer cells.Bioinformatics software was used to predict miRNAs targeting SRGN,and integrated microRNA differentially expressed chip data between breast cancer cell MCF-7 versus MCF-7/5-Fu.The miRNA quantitative PCR was used to determine the differences of candi date miRNA expression.After transfection of microRNA minics,Western blot was used to test candidate microRNA target SRGN.Transwell experiment was used to test the effects of candidate microRNAs on tumor cell invasion and metastasis.Results SRGN was increased significantly in MCF-7/5-Fu cells,and the invasion and metastasis of tumor cells were inhibited when SRGN was interfered.In addition,miR181 b/c expressed in MCF-7/5-Fu cells was reduced significantly,negatively correlated with SRGN expression,and targeted SRGN expression.It inhibited invasion and metastasis of tumor cells.Conclusions MicroRNA181b/c inhibits metastasis of breast cancer by targeting SRGN.

Application value of combined detection of AFP-L3,GP73 and GPC3 in diagnosing primary hepatic cancer / 国际检验医学杂志

Objective To study the clinical value of the combination detection of serum tumor markers alpha fetal protein hetero-geneity-L3(AFP-L3) ,Golgi protein 73(GP73) and phosphatidylinositol proteoglycan-3(GPC-3) in the diagnosis of primary hepatic cancer .Methods The serum AFP-L3 ,GP73 and GPC-3 levels were measured in 34 patients with primary hepatic cancer (PHC) ,20 patients with liver cirrhosis ,20 patients with chronic hepatitis B ,37 patients with other tumors by the enzyme linked immunosor-bent assay(ELISA) .Meanwhile ,20 individuals with healthy physical examination were selected as the control group .The expression situation of various indexes were compared and analyzed .Results The expression levels of AFP-L3 ,GP73 and GPC-3 in the PHC group were significantly higher than those in the control group and the other tumors groups ,difference was statistically significant (P<0 .05) .The areas under the receiver operating characteristic (ROC) curve of PHC in the single item detection of 3 markers were 0 .909 ,0 .832 and 0 .817 respectively .The areas of the ROC curve in the combination examinations of two items were 0 .935 , 0 .945 and 0 .912 respectively .The area of the ROC curve in the 3-item combined detection of PHC could be up to 0 .960 .Conclusion The combined detection of AFP-L3 ,GP73 and GPC-3 can increase the detective rate of PHC and has important clinical significance to early diagnosis of PHC .