Increased expression of matrix metalloproteinases 2 and 9 as poor prognosis factor for Hodgkin's lymphoma patients

Abstract Objective: The aim of our study was to evaluate the expression of MMP-2 and MMP-9 as a prognostic factor in patients diagnosed with Hodgkin Lymphoma (HL). Methods: In the present study, 45 paraffin biopsies from patients up to 19 years old diagnosed with HL were used in two referral hospitals in the state of Pernambuco, Brazil. Risk groups were classified into favorable and unfavorable, according to Ann Arbor. The expression of matrix metalloproteinases 2 and 9 and their inhibitors was performed by immunohistochemistry (IHC). Data were analyzed using the GraphPad Prism 5 program. Results: MMP-2 intensity pattern was stronger (> 10% of the total field) in patients with stage III/ IV and B symptoms. MMP-2 showed an association with the risk group (p = 0.0388). That is, the stronger the MMP-2 marking, the greater the unfavorable risk. However, for MMP-9 there was no difference in the stronger intensity pattern in relation to stages I/II and III/IV, only in the presence of B symptoms. MMP-9 showed an association with B Symptoms (p = 0.0411). Therefore, patients with B symptoms have higher MMP-9 expression. Conclusion: Our results suggest that MMP-2 expression is associated with HL progression. While MMP-9 expression is related to the clinical worsening of these patients. However, further studies are needed to evaluate the exact role of these proteins in hematologic malignancies.

Application of proanthocyanidin in the treatment and repair of deep caries / 口腔疾病防治

@#Proanthocyanidin (PA), as a kind of natural plant polyphenol, have a variety of biological functions, such as promoting remineralization, inducing collagen cross-linking, inhibiting protease activity and inhibiting bacteria. Therefore, PA could be broadly used in the clinical application of treatment and repair of deep caries in the future; for example, PA could promote dentin remineralization, improve resin-dentin bonding durability and improve the dentin acid erosion effect. This application potential of PA arises from several features, firstly, PA can not only promote dentin remineralization on its own or with other remineralizers but also exhibits antibacterial effects, which can inhibit acid production while reducing the formation of cariogenic pathogens and their biofilms. Based on the above features, PA can reduce the incidence of caries disease; thus, PA improves deep caries and long-term effects after treatment. In addition, PA added to adhesives or etch agents can improve the etching and bonding effect of dentin by inducing collagen cross-linking and inhibiting protease activity, thus achieving the ultimate goal of improving the bonding performance of deep caries. This paper summarizes recent progress of research on PA for the treatment and repair of deep caries, including the promotion of dentin remineralization and antibacterial activity as well as the improvement in dentin bonding and acid etching effect, to provide a more comprehensive reference for treating and restoring deep caries in clinical practice.

The Effect of Low Tidal Volume Ventilation on Inflammatory Cytokines During Cardiopulmonary Bypass

Abstract Introduction: Halting ventilation during cardiopulmonary bypass (CPB) is implemented to operate in a less bleeding setting. It sustains a better visualization of the operation area and helps to perform the operation much more comfortably. On the other hand, it may lead to a series of postoperative lung complications such as atelectasis and pleural effusion. In this study, we investigated the effects of low tidal volume ventilation on inflammatory cytokines during CPB. Methods: Twenty-eight patients undergoing cardiovascular surgery were included in the study. Operation standards and ventilation protocols were determined and patients were divided into two groups: patients ventilated with low tidal volume and non-ventilated patients. Plasma samples were taken from patients preoperatively, perioperatively from the coronary sinus and postoperatively after CPB. IL-6, IL-8, TNF-α and C5a levels in serum samples were studied with enzyme-linked immunosorbent assay (ELISA) kits. Results: C5a, IL-6, IL-8 and TNF-α were similar when compared to the low tidal volume ventilated and non-ventilated groups (P>0.05) Comparing the groups by variables, IL-6 levels were increased during CPB in both groups (P=0.021 and P=0.001), and IL-8 levels decreased in the ventilation group during CPB (P=0.018). Conclusion: Our findings suggest that low tidal volume ventilation may reduce the inflammatory response during CPB. Although the benefit of low tidal volume ventilation in CPB has been shown to decrease postoperative lung complications such as pleural effusion, atelectasis and pneumonia, we still lack more definitive and clear proofs of inflammatory cytokines encountered during CPB.

Phosphoric acid containing proanthocyanidin enhances bond stability of resin/dentin interface

Abstract Proanthocyanidin (PA) is a promising dentin biomodifier due to its ability to stabilize collagen fibrils against degradation by matrix metalloproteinases (MMPs); however, the most effective protocol to incorporate PA into bonding procedures is still unclear. This study evaluated the effect of dentin biomodification with a PA acid etchant on MMP activity, adhesive interface morphology and resin-dentin microtensile bond strength. Sound extracted human molars were flattened to expose dentin and acid-etched for 15 s according to the groups: EXP - experimental phosphoric acid; EXP+PA - experimental phosphoric acid 10% PA; TE - total-etching system; SE - self-etching system. Samples were restored with composite resin and stored in distilled water (37ºC). MMP activity and interface morphology were analyzed after 24 h by in situ zymography (n=6) and scanning electron microscopy (n=3), respectively. The resin-dentin microtensile bond strength (μTBS) was evaluated after 24 h and 6 months storage (n=6). Significantly higher MMP activity was detected in etched dentin compared with untreated dentin (p<0.05), but no difference among acid groups was found. Resin tags and microtags, indicative of proper adhesive system penetration in dentinal tubules and microtubules, were observed along the hybrid layer in all groups. There was no difference in μTBS between 24 h and 6 months for EXP+PA; moreover, it showed higher long-term μTBS compared with TE and EXP (p<0.05). The results suggest that 15 s of biomodification was not sufficient to significantly reduce MMP activity; nonetheless, EXP+PA was still able to improve resin-dentin bond stability compared with total- and self-etching commercial systems.

Resumo A proantocianidina (PA) é um biomodificador dentinário promissor devido a sua capacidade de estabilizar as fibrilas colágenas contra a degradação por metaloproteinases da matriz (MMPs); no entanto, o protocolo mais eficaz para a incorporação de PA em procedimentos adesivos ainda não está claro. Este estudo avaliou o efeito da biomodificação da dentina com um condicionador ácido contendo PA na atividade de MMPs, morfologia da interface adesiva e resistência à microtração resina-dentina. Molares humanos extraídos foram lixados para exposição da dentina e condicionados com ácido por 15 s de acordo com os grupos: EXP - ácido fosfórico experimental; EXP+PA - ácido fosfórico experimental com 10% PA; TE - sistema total-etch; SE - sistema self-etch. As amostras foram restauradas com resina composta e armazenadas em água destilada (37ºC). A atividade de MMP e morfologia da interface foram analisadas após 24 h por zimografia in situ (n=6) e microscopia eletrônica de varredura (n=3), respectivamente. A resistência à microtração resina-dentina (μTBS) foi avaliada após 24 horas e 6 meses de armazenamento (n=6). Atividade de MMP detectada na dentina condicionada foi significativamente maior em comparação com a dentina não tratada (p <0,05), mas não houve diferenças entre os diferentes ácidos. Tags e microtags de resina, indicativos de uma penetração adequada do sistema adesivo nos túbulos e microtúbulos dentinários, foram observadas ao longo da camada híbrida em todos os grupos. Não houve diferença entre os valores de μTBS de 24 h e 6 meses para EXP+PA; além disso, EXP+PA apresentou maiores valores de μTBS após 6 meses em comparação com TE e EXP (p <0,05). Os resultados sugerem que a biomodificação por 15 s não foi suficiente para reduzir significativamente a atividade de MMP; apesar disso, EXP + PA foi capaz de melhorar a estabilidade da interface resina-dentina em comparação com sistemas total- e self-etch comerciais.

Perfil inflamatório e microbiológico de pacientes com infecção endodôntica primária e periodontite apical: estudo observacional / Inflammatory and microbiological profile of patients with primary endodontic infection and apical periodontitis: observational study

O objetivo deste estudo foi avaliar o perfil microbiológico e identificar padrões de agregação bacteriana, avaliar a associação desses padrões com os níveis de mediadores inflamatórios, MMPs e sinais e sintomas clínicos e ainda correlacionar os mediadores inflamatórios entre si e com os sinais e sintomas clínicos em dentes com infecção endodôntica primária e periodontite apical (PA). Para isso, 40 dentes uniradiculares, de pacientes com infecção endodôntica primária e PA, foram submetidos à avaliação clínica, com registro de sinais e sintomas, e tomográfica através do cálculo de volumetria das lesões periapicais com o software ITK Snap. Em seguida os dentes foram submetidos ao tratamento endodôntico. Logo após a abertura coronária, amostras foram coletadas de cada canal radicular utilizando cones de papel (S1) e submetidas ao método Checkerboard DNA-DNA hybridization para investigação de espécies bacterianas presentes. Foi realizado o preparo biomecânico (PBM) e em seguida o fluído intersticial da PA foi coletado (SF1) para a quantificação dos mediadores inflamatórios (IL1-ß, IL-6, TNF-α, IL-10, MMPs -2 e -9) através do ensaio multiplex e a quantificação de RvD2 através de ensaio imunoenzimático ELISA. Para a análise de correlação o teste de correlação de Spearman foi utilizado. A análise fatorial foi usada para identificar padrões de agregação bacteriana e regressão linear foi realizada para associar os escores fatoriais, mediadores e características clínicas dos pacientes (P=0,05). Correlação positiva foi encontrada entre MMP-2, -9, IL-10, IL-1ß, IL-6 e TNF-α, e entre RvD2, MMP-9, IL-10, IL-1ß, IL-6 e TNF-α (P<0,05), e correlação negativa foi encontrada entre IL-1ß e sensibilidade a percussão (P<0,05). A análise microbiológica revelou presença de DNA bacteriano em 100% das amostras analisadas com presença de pelo menos 2 das 40 espécies bacterianas investigadas (média = 24,62) por canal. As espécies mais frequentemente detectadas foram P. gingivalis, E. nodatum, F. nucleatum spp. vicentii, S. mitis, L. bucallis e A. actinomycetemcomitans. Das 6 espécies mais detectadas, 4 delas eram gram negativas, destacando o predomínio de gram-negativos das infecções endodônticas primárias com PA. A análise fatorial determinou 2 padrões de associação bacteriana e os resultados de regressão para o fator 1 revelaram uma associação com aumento de dor a percussão (coeficiente (coef) ß=0,788) e redução de dor a palpação (coefß=-0,753) e exsudato (coefß=- 0,479). Níveis mais altos de exsudato e menores de dor a palpação foram associados a bactérias do fator 2 (coefß=0,460 e -0,546, respectivamente). Concluímos que os mediadores inflamatórios formam uma rede inter-relacionada e que as periodontites apicais assintomáticas e sintomáticas tem uma etiologia bacteriana heterogênea e com combinações de espécies diferentes (AU)

The aim of this study was to evaluate the microbiological profile and identify patterns of bacterial aggregation, evaluate the association of these patterns with the levels of inflammatory mediators, MMPs and clinical signs and symptoms, and also correlate the inflammatory mediators with each other and with the clinical signs and symptoms in teeth with primary endodontic infection and apical periodontitis (AP). For this, 40 single-rooted teeth, from patients with primary endodontic infection and PA, were submitted to clinical evaluation, with registration of signs and symptoms, and tomography through the calculation of volumetry of periapical lesions with the ITK Snap software. Then the teeth were submitted to endodontic treatment. Immediately after the coronal opening, samples were collected from each root canal using paper cones (S1) and submitted to the Checkerboard DNA-DNA hybridization method to investigate bacterial species present. Biomechanical preparation (BMP) was performed and then the AP interstitial fluid was collected (SF1) for the quantification of inflammatory mediators (IL1-ß, IL-6, TNF-α, IL-10, MMPs -2 and -9) by the multiplex assay and the quantification of RvD2 by enzyme immunoassay ELISA. For correlation analysis, Spearman's correlation test was used. Factor analysis was used to identify bacterial aggregation patterns and linear regression was performed to associate factor scores, mediators and patients' clinical characteristics (P=0.05). Positive correlation was found between MMP-2, -9, IL-10, IL-1ß, IL-6 and TNF-α, and between RvD2, MMP-9, IL-10, IL-1ß, IL-6 and TNF- α (P<0.05), and a negative correlation was found between IL-1ß and sensitivity to percussion (P<0.05). The microbiological analysis revealed the presence of bacterial DNA in 100% of the samples analyzed with the presence of at least 2 of the 40 bacterial species investigated (mean = 24.62) per root canal. The most frequently detected species were P. gingivalis, E. nodatum, F. nucleatum spp. vicentii, S. mitis, L. buccallis and A. actinomycetemcomitans. Of the 6 most detected species, 4 of them were gram negative, highlighting the predominance of gram-negative primary endodontic infections with AP. Factor analysis determined 2 patterns of bacterial association and regression results for factor 1 revealed an association with increased pain on percussion (coefficient (coef) ß=0.788) and reduced pain on palpation (coefß=-0.753) and exudate (coefß=-0.479). Higher levels of exudate and lower levels of pain on palpation were associated with factor 2 bacteria (coefß=0.460 and -0.546, respectively). We conclude that inflammatory mediators form an interrelated network and that asymptomatic and symptomatic apical periodontitis has a heterogeneous bacterial etiology and combinations of different species (AU)

The polymorphism of metalloproteinases 1 and 13 and posttraumatic elbow stiffness / O polimorfismo das metaloproteinases 1 e 13 e a rigidez articular pós-traumática do cotovelo

ABSTRACT Introduction To evaluate the relationship between the genetic polymorphism of matrix metalloproteinases 1 and 13 and posttraumatic elbow stiffness, as well as the association of other risk factors with this condition. Materials and methods We evaluated 20 patients with posttraumatic elbow stiffness and 12 controls with traumatic elbow disorders without contracture. Deoxyribonucleic acid (DNA) was obtained from buccal mucosa epithelial cells of the volunteers. The MMP-1 and MMP-13 genotypes were determined using PCR-restriction fragment length polymorphism assays. Results We did not find any significant differences in the frequency of genotypes and alleles between the test and control groups for the polymorphism of metalloproteinases 1 and 13. We observed that genotypes 1G/2G and 2G/2G of MMP-1 were present in 65% (13/20) of patients with articular stiffness and 50% (6/12) of controls (p = 0.599). Genotypes A/A and A/G of MMP-13 were obtained in 95% (19/20) of patients and 91.6% (11/12) of controls (p = 0.491). Among the prognostic factors for elbow stiffness, only immobilization time correlated positively. The mean immobilization time for cases and controls were 16 ± 10 days and 7 ± 7 days, respectively (p = 0.017). Conclusion The genetic polymorphism of MMP-1 at position -1607 and MMP-13 at position -77 was not associated with post-traumatic elbow stiffness. Level of Evidence III; Prognosis Study; Case-Control Study.

RESUMO Introdução Avaliar a relação entre o polimorfismo genético das metaloproteinases 1 e 13 da matriz e a rigidez pós-traumática do cotovelo, assim como a associação de outros fatores de risco com essa condição. Material e método Foram avaliados 20 pacientes com rigidez pós-traumática do cotovelo e 12 controles com distúrbios traumáticos do cotovelo sem contratura. O ácido desoxirribonucleico (DNA) de voluntários foi obtido a partir de células epiteliais da mucosa bucal. Os genótipos MMP-1 e MMP-13 foram determinados usando ensaios de polimorfismo de comprimento de fragmento de restrição de PCR. Resultados Não encontramos diferença significativa na frequência de genótipos e alelos entre os grupos teste e controle para o polimorfismo das metaloproteinases 1 e 13. Observamos que os genótipos 1G/2G e 2G/2G de MMP-1 estavam presentes em 65% (13/20) dos pacientes com rigidez articular e 50% (6/12) dos controles (p = 0,599). Os genótipos A/A e A/G da MMP-13 foram obtidos em 95% (19/20) dos pacientes e 91,6% (11/12) dos controles (p = 0,491). Dentre os fatores prognósticos para rigidez de cotovelo, apenas o tempo de imobilização se correlacionou positivamente. O tempo médio de imobilização para casos e controles foi de 16 ± 10 dias e 7 ± 7 dias, respectivamente (p = 0,017). Conclusões O polimorfismo genético de MMP-1 na posição -1607 e MMP-13 na posição -77 não foi associado à rigidez pós-traumática do cotovelo. Nível de Evidência III; Estudos Prognósticos; Estudo de Caso-Controle.

Regulation of liver fibrosis by matrix metalloproteinase/tissue inhibitor of metalloproteinase and research advances in related therapeutic drugs / 临床肝胆病杂志

Liver fibrosis is the common consequence of various chronic liver injuries and is mainly characterized by the imbalance between the production and degradation of extracellular matrix, which leads to the accumulation of interstitial collagen and other matrix components. Matrix metalloproteinases (MMPs) and their specific inhibitors, i.e., tissue inhibitors of metalloproteinases (TIMPs), play a crucial role in collagen synthesis and lysis. Through a literature review, this article reviews the experimental studies of liver fibrosis based on MMPs/TIMPs, summarizes the components that may exert an anti-liver fibrosis effect by affecting the expression or activity of MMPs/TIMPs, and attempts to clarify the mechanism of MMPs/TIMPs in regulating collagen homeostasis, so as to provide support for the development of anti-liver fibrosis drugs.

Progress in the study of intracranial arterial dolichoectasia / 中华神经科杂志

Intracranial arterial dolichoectasia (IADE), also known as dilated cerebral artery disease, is manifested as an increase in the length and diameter of one or more intracranial arteries, the affected arteries being enlarged and dilated significantly, or even with winding and tortuosity. The diagnosis and prognostic evaluation of IADE are mainly based on the diameter and curvature of the intracranial artery. IADE can be manifested as ischemic stroke, transient ischemic attack, hydrocephalus, and hemorrhagic stroke. The review focuses on IADE, including the concept, diagnostic criteria, etiology, pathogenesis, pathology, clinical manifestations, treatment, and prognosis.

Effect of propofol on proliferation, invasion and migration of human melanoma cells and role of COX-2/PGE2/MMP signaling pathway / 中华麻醉学杂志

Objective:To evaluate the effect of propofol on proliferation, invasion and migration of human melanoma cells and role of cyclooxygenase-2 (COX-2)/prostaglandin E2 (PGE2)/matrix metalloproteinase (MMP) signaling pathway.Methods:SKMEL-5 cells were cultured in vitro and divided into 4 groups ( n=36 each) using the random number table method: control group (group C), propofol group (group P), COX-2 overexpression group (group COX-2), and COX-2 overexpression plus propofol group (group COX-2+ P). Propofol at the final concentration of 60 μmol/L was added in group P. The COX-2 overexpression plasmid pcDNA3.1-COX-2 was transfected into SKMEL-5 cells in group COX-2 and group COX-2+ P, and propofol at the final concentration of 60 μmol/L was added in group COX-2+ P.After incubation for 48 h, the cell proliferation rate was determined by CCK-8 method, the cell invasion and migration ability was determined by Transwell assay, the expression of COX-2 in cells was detected by Western blot, the expression of COX-2 mRNA in cells was detected by quantitative real-time polymerase chain reaction, and the concentrations of serum PGE2, MMP-2 and MMP-9 were determined by enzyme-linked immunosorbent assay. Results:Compared with group C, the cell proliferation rate was significantly decreased, the number of cell invasion and migration was decreased, the expression of COX-2 protein and mRNA was down-regulated, and the concentrations of PGE2, MMP-2 and MMP-9 in the supernatant were decreased in group P, and the cell proliferation rate was significantly increased, and the number of cell invasion and migration was increased, the expression of COX-2 protein and mRNA was up-regulated, and the concentrations of PGE2, MMP-2 and MMP-9 in the supernatant were increased in group COX-2 ( P<0.05). Compared with group P, the cell proliferation rate was significantly increased, and the number of cell invasion and migration was increased, the expression of COX-2 protein and mRNA was up-regulated, and the concentrations of PGE2, MMP-2 and MMP-9 in the supernatant were increased in group COX-2+ P ( P<0.05). Conclusions:Propofol can inhibit the proliferation, invasion and migration of human melanoma cells, and the mechanism may be related to inhibition of the COX-2/PGE2/MMP signaling pathway.

Research progress on the factors influencing the bonding effect of universal adhesives on tooth tissues / 口腔疾病防治

@#Universal adhesives have been widely used in dentistry due to their easy application process and wide range of applications. In the literature, the etching modes of universal adhesives, coating methods of universal adhesives, pretreatment of adhesive, and other factors were reported to have an impact on the bonding performance of universal adhesives. This review focused on the factors affecting the bonding performance of universal adhesives and aimed to provide evidence-based recommendations for clinical practice. Current research suggests that the etch-and-rinse mode can achieve a better adhesive strength of enamel and that the etch-and-rinse mode and the self-etch mode exhibit a similar adhesive performance in bonding dentin. The bond strength would be improved by prolonging the application time, applying a double layer of adhesives, and wetting enamel and dentin with water or ethanol before adhesive procedures. Chlorohexidine (0.2%) and metal nanoparticles can inhibit matrix metalloproteinases (MMPs) and improve the long-term bond strength of dentin. Finally, universal adhesives have excellent stability and saliva pollution resistance, although they lack blood pollution resistance. In the clinic, dentists should pay attention to the etching modes of universal adhesives, coating methods of universal adhesives, pretreatment of adhesive, and other factors that may affect the effect strength. universal adhesives will be improved in bonding strength of enamel in self-etch mode and bonding performance in demineralized dentals around caries.

Efeito do tratamento com antileucotrieno em contratura capsular: estudo experimental / Effect of antileukotriene treatment on capsular contracture: an experimental study

RESUMO Introdução: 40% dos pacientes submetidos à radioterapia após reconstrução de mama por implante de prótese de silicone podem desenvolver encapsulamento da prótese. Diversas estratégias já foram testadas para prevenir a contratura da cápsula com resultados insatisfatórios. Este estudo analisou o efeito do antileucotrieno (AL) tópico na formação de contratura capsular em ratos com implantes de silicone associados à irradiação. Métodos: Foram implantados blocos de silicone na região dorsal em 20 ratas fêmeas, espécie Wistar com peso variando de 200-250g. Os animais foram divididos em dois grupos: controle (injeção de solução fisiológica 0,9% no tecido ao redor do implante) e grupo intervenção (injeção de 10mg de AL no tecido ao redor do implante). Imediatamente após a cirurgia os animais foram irradiados com dose única de 10Gy. Após dois meses, coletamos amostras de cápsulas para análise histológica e análise da expressão gênica dos seguintes biomarcadores: iNOS, VEGF-a e MMP-9. Resultados: A densidade vascular foi menor no grupo AL quando comparado ao grupo controle (55,4±30,0 vs. 81,8±26,7, p=0,05, respectivamente). Da mesma forma, o VEGF-a teve o mesmo comportamento (grupo controle - 0,34±0,1 vs. grupo Al - 0,02±0,001, p=0,04). Conclusão: Este estudo sugeriu que o tratamento com AL diminui a angiogênese em animais submetidos a implantes de silicone e submetidos à radioterapia

ABSTRACT Introduction: 40% of patients undergoing radiotherapy after breast reconstruction by silicone prosthesis implant may develop prosthesis encapsulation. Several strategies have already been tested to prevent capsule contracture with unsatisfactory results. This study analyzed the effect of topical antileukotriene (AL) on capsular contracture formation in rats with silicone implants associated with irradiation. Methods: Silicone blocks were implanted in the dorsal region in 20 female rats Wistar with weights ranging from 200-250g. The animals were divided into two groups: control (injection of 0.9% saline solution into the tissue around the implant) and intervention group (injection of 10mg of AL into the tissue around the implant). Immediately after surgery, the animals were irradiated with a single dose of 10Gy. After two months, we collected capsule samples for histological analysis and gene expression analysis of the following biomarkers: iNOS, VEGF-a and MMP-9. Results: Vascular density was lower in the AL group when compared to the control group (55.4±30.0 vs. 81.8±26.7, p=0.05, respectively). Similarly, VEGF-a had the same behavior (control group - 0.34±0.1 vs. group Al - 0.02±0.001, p=0.04). Conclusion: This study suggested that treatment with AL decreases angiogenesis in animals submitted to silicone implants and underwent radiotherapy.

Expression of metalloproteinases 2 and 9 and plasma zinc concentrations in women with fibroadenoma

SUMMARY OBJECTIVE: This study aims to investigate the association between the immunohistochemical expression of matrix metalloproteinase-2 and matrix metalloproteinase-9 and plasma zinc in women with fibroadenoma. METHODS: This cross-sectional study included 37 premenopausal women with fibroadenoma. Waist circumference and body mass index of the participants were measured. Plasma zinc concentrations were determined using atomic flame absorption spectrophotometry. Fragments of breast tissue were fixed and incubated with primary mouse monoclonal antibodies (monoclonal antibodies matrix metalloproteinase -2 -507 and monoclonal antibodies matrix metalloproteinase -9-439). Semi-quantitative analysis of matrix metalloproteinase-2 and matrix metalloproteinase-9 immunoreactivity was performed. Spearman's test and Friedman's test were used for statistical analyses. The p<0.05 were considered statistically significant. RESULTS: The average age of the participants was 32.81±9.51 years. The body mass index and waist circumference values were within the normal range. The mean plasma zinc concentration was 42.73±13.84 µg/dL, with 94.6% inadequacy. A statistically significant difference was found between the positive expression of matrix metalloproteinase-2 and matrix metalloproteinase-9 (p=0.0184). There was no significant correlation between the matrix metalloproteinase expression and the plasma zinc levels. CONCLUSIONS: Women with fibroadenoma had hypozincemia and positive expression of metalloproteinases.

The study of Yangyinyiqi mixture anti bleomycin-induced pulmonary fibrosis on rats by intervening matrix metalloproteinase-9 and tissue inhibitors of matrix metallopreoteinases-1 / El estudio de la mezcla Yangyinyiqi en fibrosis pulmonar inducida por anti-bleomicina en ratas por intervención en la matriz metaloproteinasa-9 e inhibidores de tejido de matriz metaloproteinasa-1

To investigate effectsof Yangyinyiqi Mixture on pulmonary fibrosis caused by bleomycin. SD ratswere divided randomly into: model group(distilled water,1 mL·0.1 kg-1), dexamethasone acetate group (dexamethasone acetate, the dosage was reduced gradually), low-dose group (Yangyinyiqi Mixture, 11 g·kg-1), moderate-dose group (Yangyinyiqi Mixture, 22 g·kg-1), high-dose group (Yangyinyiqi Mixture, 44 g·kg-1) and control group (distilled water, 1 mL·0.1 kg-1). Yangyinyiqi Mixture and dexamethasone acetate were intragastrically administrated. Lung tissue was collected for histopathological examination. Compared with control group, collagen markedly increased and HYP content significantly increased on 7th day in model group (p<0.01). On 28th day, collagen was diffusely deposited, alveolar was destroyed, and HYP content significantly increased (p<0.01). Compared with model group, bleomycin-induced suffering injury caused MMP-9 expression levels to rapidly increase (7and 14 days, p<0.01). TIMP-1 markedly increased (7and 14 days, p<0.01) and stayed at a high level to28th day. Yangyinyiqi Mixture exerted an effect against pulmonary fibrosis, which could involved prevention of collagen deposition through inhibitingMMP-9 and TIMP-1 expression.

El trabajo investiga los efectos de la mezcla Yangyinyiqi sobre la fibrosis pulmonary causada por bleomicina. Ratas SD se dividieron aleatoriamente en: grupo modelo (agua destilada, 1 mL·0.1 kg-1), grupo acetate de dexametasona (acetate de dexametasona, la dosis se redujo gradualmente), grupo de dosis baja (mezcla Yangyinyiqi, 11 g·kg-1), grupo de dosis moderada (mezcla Yangyinyiqi, 22 g·kg-1), grupo de dosis alta (mezcla Yangyinyiqi, 44 g·kg-1) y grupo control (agua destilada, 1 Ml·0.1 kg-1). La mezcla de Yangyinyiqi y el acetate de dexametasona se administraron por vía intragástrica. Se recolectó tejido pulmonary para examen histopatológico. En comparación con el grupo control, el colágeno aumentó notablemente y el contenido de HYP aumentó significativamente el séptimo día en el grupo modelo (p<0.01). El día 28, el colágeno se depositó difusamente, se produjo destrucción alveolar y el contenido de HYP aumento significativamente (p<0.01). En comparación con el grupo modelo, la lesión inducida por bleomicina causó que los niveles de expression de MMP-9 aumentaron rápidamente (7 y 14 días, p<0.01). TIMP-1 aumentó notablemente (7 y 14 días, p<0.01) y se mantuvo en un nivel alto hasta el día 28. La mezcla Yangyinyiqi ejerció un efecto contra la fibrosis pulmonary, lo que podría implicar la prevención del deposito de colágenio mediante la inhibición de la expression de MMP-9 y TIMP-1.

Proteolytic activity and degradation of bovine versus human dentin matrices

Abstract Non-human teeth have been commonly used in research as replacements for human teeth, and potential dissimilarities between the dental tissues should be considered when interpreting the outcomes. Objective: To compare the proteolytic activity and degradation rate of bovine and human dentin matrices. Methodology: Dentin beam specimens were obtained from human molars (n=30) and bovine incisors (n=30). The beams were weighed hydrated and after complete dehydration to obtain the mineralized wet and dry masses. Then, the beams were demineralized in 10 wt% phosphoric acid. Next, 15 beams from each substrate were randomly selected and again dehydrated and weighed to obtain the initial demineralized dry mass (DM). Then, the beams were stored in saliva-like buffer solution (SLBS) for 7, 14 and 21 days. SLBS was used to evaluate hydroxyproline (HYP) release after each storage period. The remaining beams of each substrate (n=15) were tested for initial MMP activity using a colorimetric assay and then also stored in SLBS. DM and MMP activity were reassessed after 7, 14 and 21 days of incubation. The data were subjected to two-way ANOVA tests with repeated measures complemented by Bonferroni's tests. Unpaired two-tailed t-tests were also used (p<0.05). Results: Similar water and inorganic fractions were found in human and bovine dentin, while human dentin had a higher protein content. The most intense proteolytic activity and matrix deterioration occurred short after dentin was demineralized. Both substrates exhibited a sharp reduction in MMP activity after seven days of incubation. Although human dentin had higher MMP activity levels, greater HYP release and DM loss after seven days than bovine dentin, after 14 and 21 days, the outcomes were not statistically different. Conclusion: Bovine dentin is a suitable substrate for long-term studies involving the degradation of dentin matrices.

Polymorphisms of matrix metalloproteinase-7 and -9 are associated with oral tongue squamous cell carcinoma

Abstract Matrix degradation is an important event in the progression, invasion and metastasis of malignant head and neck lesions. Imbalances, mutations and polymorphisms of MMPs and their inhibitors are observed in several cancer subtypes. The aim of this study was to evaluate the association of the MMP-7 gene promoter (181 A/G) and MMP-9 (-1562 C/T) polymorphisms in oral tongue squamous cell carcinoma (OTSCC). MMP-7 (rs11568818) and MMP-9 (rs3918242) single-nucleotide polymorphisms (SNPs) were genotyped by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis in 71 cases of OTSCC. Normal tissue specimens were obtained from 60 healthy volunteers to serve as the control. The MMP-7 G allele and MMP-9 T allele were more frequent in the OTSCC group than the control group, but only when these two SNPs were taken together was a significant association found with the nodal metastasis of OTSCC (p < 0.001). Based on our results, SNPs in the promoter region of MMP-7 and MMP-9 appear to be associated with greater risk of developing OTSCC, and with a higher propensity to form metastatic tumors. In this respect, molecular studies investigating polymorphisms may be useful in predicting tumor behavior.

Análise in vitro da capacidade de soluções contendo fluoreto e/ou trimetafosfato na remineralização da dentina e na inibição da atividade de metaloproteinases da matriz dentinária / In vitro analysis of the capacity of solutions containing fluoride and/or trimetaphosphate in the remineralization of dentin and in the inhibition of the activity of dentin matrix metalloproteinases

Objetivo: O objetivo deste estudo foi investigar o potencial anti-proteolítico contra MMP-2 e MMP-9 e a capacidade de induzir a remineralização da dentina, através de soluções contendo diferentes concentrações de TMP (na sua forma cíclica), diferentes concentrações de F e associação TMP/F. Métodos: Blocos de dentina radicular (6mmx4mmx2mm, n = 130) foram selecionados e divididos aleatoriamente em 13 grupos/soluções experimentais (n = 10): 1) Placebo (sem F/TMP); 2) 0,3% TMP hidrolisado; 3) 1% TMP hidrolisado; 4) 3% TMP hidrolisado; 5) 0,3% TMP; 6) 1% TMP; 7) 3% TMP; 8) 250 ppm F; 9 500 ppm F; 10) 1100 ppm F; 11) 250 ppm F associado a 0,3% TMP; 12) 500 ppm F associado a 1% TMP e 13) 1100 ppm F associado a 3% TMP. A avaliação do potencial anti-proteolítico das soluções experimentais contra as metaloproteinases da matriz dentinária (- 2 e -9) foi realizado por meio da análise zimográfica. Para as análises mecânicas, três áreas foram determinadas para cada espécime: 1- controle (sem tratamento); 2- desmineralizado (cárie artificial); 3- tratado (desmineralizado e submetido a ciclagem de pH por 7 dias, e tratado por 1 min com as soluções experimentais). Os espécimes de dentina foram analisados quanto à porcentagem de recuperação de dureza superficial (%SHR), em dureza transversal (%CSHR) e por microtomografia computadorizada (IMC). Para os dados de dureza e Micro-CT, os resultados foram analisados por ANOVA de medidas repetidas seguida do teste de Student-Newman-Keuls (p <0,05). Resultados: A análise zimográfica mostrou que 1100 ppm F + 3% de TMP promoveu inibição completa da atividade gelatinolítica (MMP-2; MMP-9). Os grupos com TMP não hidrolisados apresentaram efeito remineralizador (% SHR e % CSHR) superior aos grupos hidrolisados. O grupo 1100F + TMP promoveu a maior %SHR e %CSHR entre todos os grupos (p <0,001), sendo respectivamente 15,4 e 10,5%, superior ao grupo 1100F. Os grupos contendo 1100F e 1100F + 3%TMP apresentaram maior concentração mineral. Conclusão: Com base nos resultados, 3% TMP atua como um agente antiproteolítico contra metaloproteinases da matriz dentinária. Além disso, quando suplementado com 1100F, 3% TMP potencializa a remineralização, aumentando significativamente as propriedades mecânicas da dentina tratada. Os tratamentos com TMP não hidrolisado e associado ao F maior que as soluções fluoretadas sem o TMP. Significância clínica: Dessa forma, uma potencial estratégia preventiva e terapêutica pode ser considerada na clínica odontológica, principalmente na terapia da cárie radicular, como também para método preventivo de lesões iniciais de cárie dentinária ou no pré-tratamento da dentina, utilizando-os como agentes cross-linker em procedimentos restauradores(AU)

Objective: The aim of this study was to investigate the anti-proteolytic potential against MMP-2 and MMP-9 and the ability to induce dentin remineralization, through solutions containing different concentrations of TMP (in its cyclic form), different concentrations of F and TMP / F association. Methods: Root dentin blocks (6mmx4mmx2mm, n = 130) were selected and randomly divided into 13 groups / experimental solutions (n = 10): 1) Placebo (without F / TMP); 2) 0.3% hydrolyzed TMP; 3) 1% hydrolyzed TMP; 4) 3% hydrolyzed TMP; 5) 0.3% TMP; 6) 1% TMP; 7) 3% TMP; 8) 250 ppm F; 9,500 ppm F; 10) 1100 ppm F; 11) 250 ppm F associated with 0.3% TMP; 12) 500 ppm F associated with 1% TMP and 13) 1100 ppm F associated with 3% TMP. The evaluation of the antiproteolytic potential of experimental solutions against dentin matrix metalloproteinases (- 2 and -9) was carried out by means of zymographic analysis. For mechanical analysis, three areas were determined for each specimen: 1- control (without treatment); 2- demineralized (artificial caries); 3- treated (demineralized and subjected to pH cycling for 7 days, and treated for 1 min with the experimental solutions). The dentin specimens were analyzed for the percentage of recovery of superficial hardness (% SHR), in crosssectional hardness (% CSHR) and by computed microtomography (∆IMC). For the hardness and Micro-CT data, the results were analyzed by repeated measures ANOVA followed by the Student-Newman-Keuls test (p <0.05). Results: The zymographic analysis showed that 1100 ppm F + 3% TMP promoted complete inhibition of gelatinolytic activity (MMP-2; MMP-9). The groups with non-hydrolyzed TMP showed a remineralizing effect (% SHR and % CSHR) superior to the hydrolyzed groups. The 1100F + TMP group promoted the highest % SHR and % CSHR among all groups (p <0.001), being 15.4 and 10.5%, respectively, higher than the 1100F group. The groups containing 1100F and 1100F + 3% TMP showed higher mineral concentration. Conclusion: Based on the results, 3% TMP acts as an antiproteolytic agent against dentinal matrix metalloproteinases. In addition, when supplemented with 1100F, 3% TMP enhances remineralization, significantly increasing the mechanical properties of the treated dentin. Treatments with non-hydrolyzed TMP and associated with F greater than fluoridated solutions without TMP. Clinical significance: Thus, a potential preventive and therapeutic strategy can be considered in the dental clinic, mainly in the treatment of root caries, as well as for the preventive method of initial lesions of dental caries or in the pretreatment of dentin, using them as agents cross-linker in restorative procedures(AU)

Lentinula edodes extract inhibits matrix metalloproteinase expression and increases typeⅠprocollagen expression via the p38 MAPK/c-Fos signaling pathway in ultraviolet A and B-irradiated HaCaT keratinocytes

Objective:To determine the effect of Lentinula edodes extract on ultraviolet (UV) A and UVB-induced changes in matrix metalloproteinase (MMP) and typeⅠprocollagen expression using human immortalized HaCaT keratinocytes. Methods:Lentinula edodes ethanol extract (LEE) was obtained by extraction with 80％ ethanol for 4 h at 80 ℃. Effect of LEE on UV-induced alteration on the expression and production of MMPs and typeⅠprocollagen in keratinocytes was investigated using ELISA, RT-PCR, and Western blotting assay. To determine the underlying mechanism of LEE-mediated effects, mitogen-activated protein kinase (MAPK) and activator protein 1 signaling pathways were analysed by Western blotting assay. Results:LEE significantly inhibited the expression of MMP-1 and MMP-9 and increased the expression of typeⅠprocollagen in UVA and UVB-irradiated HaCaT keratinocytes. The phosphorylation levels of p38 were significantly inhibited by LEE whereas it did not affect c-Jun N-terminal kinase and extracellular signal-regulated kinase phosphorylation. Suppression of p38 phosphorylation was also accompanied by downregulation of UVA and UVB-induced increase in c-Fos. Conclusions:LEE effectively inhibits the expression of MMP-1 and MMP-9 and increases typeⅠprocollagen production through the p38 MAPK/c-Fos signaling pathway in UVA and UVB-irradiated HaCaT keratinocytes. This findings suggest that Lentinula edodes may be developed as a cosmetic material to suppress UV exposure-mediated skin aging.

P-I metalloproteinases and L-amino acid oxidases from Bothrops species inhibit angiogenesis

Snake venoms are composed of pharmacologically active proteins that are evolutionarily diverse, stable and specific to targets. Hence, venoms have been explored as a source of bioactive molecules in treating numerous diseases. Recent evidences suggest that snake venom proteins may affect the formation of new blood vessels. Excessive angiogenesis has been implicated in several pathologies including tumours, diabetic retinopathy, arthritis, inter alia. In the present study, we have examined the effects of P-I metalloproteinases isolated from Bothrops moojeni (BmMP-1) and Bothrops atrox (BaMP-1) and L-amino acid oxidases (LAAO) isolated from B. moojeni (BmLAAO) and B. atrox (BaLAAO) on biochemical and functional aspects of angiogenesis. Methods: P-I metalloproteinases and LAAO were purified from venom by molecular size exclusion and ion-exchange chromatography and subsequently confirmed using mass spectrometry. The P-I metalloproteinases were characterized by azocaseinolytic, fibrinogenolytic and gelatinase activity and LAAO activity was assessed by enzyme activity on L-amino acids. Influence of these proteins on apoptosis and cell cycle in endothelial cells was analysed by flow cytometry. The angiogenic activity was determined by in vitro 3D spheroid assay, Matrigel tube forming assay, and in vivo agarose plug transformation in mice. Results: P-I metalloproteinases exhibited azocaseinolytic activity, cleaved α and partially β chain of fibrinogen, and displayed catalytic activity on gelatin. LAAO showed differential activity on L-amino acids. Flow cytometry analysis indicated that both P-I metalloproteinases and LAAO arrested the cells in G0/G1 phase and further induced both necrosis and apoptosis in endothelial cells. In vitro, P-I metalloproteinases and LAAO exhibited significant anti-angiogenic properties in 3D spheroid and Matrigel models by reducing sprout outgrowth and tube formation. Using agarose plug transplants in mice harbouring P-I metalloproteinases and LAAO we demonstrated a marked disruption of vasculature at the periphery. Conclusion: Our research suggests that P-I metalloproteinases and LAAO exhibit anti-angiogenic properties in vitro and in vivo.(AU)

Research progress of urine TIMP-2×IGFBP-7 in early diagnosis, risk stratification and prognosis of acute kidney injury / 中华检验医学杂志

Acute kidney injury (AKI) is a common complication in critically ill patients with complex etiology and high morbidity, which is closely related to the patient's mortality rate, hospital stay and long-term poor outcomes. Therefore, timely detection of AKI in the early reversible stage is particularly important to prevent its progression to renal failure and initiating renal replacement therapy. Therefore, exploring the relevant biomarkers in the occurrence and development of acute kidney injury has important clinical significance for the diagnosis and treatment of the disease. Tissue inhibitor of metalloproteinases-2 (TIMP-2) and insulin-like growth factor-binding protein-7 (IGFBP-7) are used as cell cycle arrest proteins, which has shown certain advantages in the early diagnosis, risk stratification, prognosis judgment, and treatment effect of acute kidney injury. Cell cycle arrest protein [TIMP-2]×[IGFBP-7] plays a role in acute kidney injury caused by various reasons and can be used as a reference index for disease prognosis.

Evaluación de la resistencia de unión a dentina humana de un sistema adhesivo universal con clorhexidina utilizado en modo de grabado total y autocondicionante / Avaliação da resistência de união à dentina humana de um sistema adesivo universal com clorexidina utilizado no modo de condicionamento total e autocondicionante Romina Tessore1 / Human dentin bond strength of a chlorhexidine containing universal adhesive system used in total-etch and self-etch mode

Objetivos: Evaluar la resistencia de unión a la microtracción en dentina humana de un sistema adhesivo universal con clorhexidina en su composición, en modo de grabado y lavado en 2 pasos, y en modo de autograbado. Metodología: 20 terceros molares divididos aleatoriamente en 4 grupos según el tipo de sistema adhesivo utilizado (Single Bond Universal®, 3MESPE y Peak Universal Bond®, Ultradent Products) y modo de uso (grabado total y autograbado). Se confeccionaron cuerpos de prueba sometidos al ensayo de microtracción utilizando una máquina de ensayos universales. Resultados: No se encontraron diferencias estadísticamente significativas entre los cuatro grupos estudiados. Conclusiones: La resistencia de unión de una resina compuesta a dentina humana no fue afectada por el uso de un sistema adhesivo universal que contiene clorhexidina en su composición aplicado en los modos de grabado total y autograbado

Objectives: To evaluate the microtensile bond strength to human dentin of chlorhexidine containing universal adhesive system applied in the total-etch and selfetch modes. Methods: Twenty third molars were randomly divided into four groups according to the universal adhesive system (Single Bond Universal ®, 3M ESPE and Peak Universal Bond®, Ultradent) and application mode used (total-etch and self-etch). Specimens were prepared and subjected to a microtensile bond strength using a MTS SANS universal testing machine. Results: No statistically significant differences were found in the microtensile bond strength between the four groups studied. Conclusions: The bond strength of a composite resin to human dentin was not affected by the use of a universal adhesive system with chlorhexidine in its composition applied in the total-etch and self-etch modes.

Objetivos: Avaliar a resistência de união á microtração na dentina humana de um sistema adesivo universal com clorexidina na sua composição, no modo de condicionamento total em dois passos e no modo autocondicionante. Metodologia: 20 terceiros molares foram divididos aleatoriamente em 4 grupos, de acordo com o tipo de sistema adesivo utilizado (Single Bond Universal®, 3MESPE e Peak Universal Bond®, Ultradent Products) e modo de uso (condicionamento total e autocondicionante). Os corpos de prova criados foram submetidos ao teste de microtração utilizando uma máquina de ensaios universal. Resultados: Não foram encontradas diferenças estatisticamente significativas entre os quatro grupos estudados. Conclusões: A resistência de união da resina composta à dentina humana não foi afetada pelo uso de um sistema adesivo universal contendo clorexidina em sua composição, aplicado nos modos de condicionamento total ou autocondicionamento.