Reduction of oxidative stress improves insulin signaling in cardiac tissue of obese mice / Redução do estresse oxidativo melhora a sinalização da insulina em tecido cardíaco de camundongos obesos

ABSTRACT Objective To evaluate the effects of oxidative stress on insulin signaling in cardiac tissue of obese mice. Methods Thirty Swiss mice were equally divided (n=10) into three groups: Control Group, Obese Group, and Obese Group Treated with N-acetylcysteine. After obesity and insulin resistance were established, the obese mice were treated with N-acetylcysteine at a dose of 50mg/kg daily for 15 days via oral gavage. Results Higher blood glucose levels and nitrite and carbonyl contents, and lower protein levels of glutathione peroxidase and phosphorylated protein kinase B were observed in the obese group when compared with their respective control. On the other hand, treatment with N-acetylcysteine was effective in reducing blood glucose levels and nitrite and carbonyl contents, and significantly increased protein levels of glutathione peroxidase and phosphorylated protein kinase B compared to the Obese Group. Conclusion Obesity and/or a high-lipid diet may result in oxidative stress and insulin resistance in the heart tissue of obese mice, and the use of N-acetylcysteine as a methodological and therapeutic strategy suggested there is a relation between them.

RESUMO Objetivo Avaliar os efeitos do estresse oxidativo sobre a sinalização da insulina em tecido cardíaco de camundongos obesos. Métodos Utilizaram-se 30 camundongos Swiss subdivididos igualmente (n=10) em três grupos: Grupo Controle, Grupo Obeso e Grupo Obeso Tratado com N-acetilcisteína. Após estabelecidas a obesidade e a resistência à insulina, os camundongos obesos foram tratados diariamente, durante 15 dias, via gavagem oral, com N-acetilcisteína na dose de 50mg/kg. Resultados Observaram-se maiores níveis de glicose sanguínea, conteúdos de nitrito e carbonil, e menores níveis proteicos de glutationa peroxidase e proteína quinase B fosforilada no Grupo Obeso quando comparado a seu respectivo controle. Por outro lado, o tratamento com N-acetilcisteína se mostrou eficiente em diminuir os níveis glicêmicos, os conteúdos de nitrito e carbonil, e aumentar significativamente os níveis proteicos de glutationa peroxidase e proteína quinase B fosforilada, quando comparados ao Grupo Obeso. Conclusão Obesidade e/ou dieta hiperlipídica levam a estresse oxidativo e à resistência à insulina no tecido cardíaco de camundongos obesos, e o uso da N-acetilcisteína como estratégia metodológica e terapêutica sugeriu haver relação entre ambos.

Losartan improves alveolar bone dynamics in normotensive rats but not in hypertensive rats

Abstract Hypertension is one of the main causes of premature death in the world; also, it is associated with several bone alterations. Preclinical studies have demonstrated delayed alveolar bone healing in hypertensive rats. However, losartan has been favorable for consolidation of bone grafts and reduction in active periodontitis. Therefore, losartan is suggested to be effective in bone formation stages, as well as in the synthesis of matrix proteins and mineralization. Objectives: To evaluate the alveolar bone dynamics in hypertensive rats treated with losartan by laser confocal microscopy and histological analysis. Methodology: Thirty-two rats, 16 spontaneously hypertensive rats (SHR) and 16 Wistar albinus rats, treated or not with losartan (30 mg/kg/day) were used. Calcein fluorochrome at 21 days and alizarin red fluorochrome at 49 days were injected in rats (both 20 mg/kg). The animals were submitted to euthanasia 67 days after treatment, and then the right maxilla was removed for laser confocal microscopy analysis and the left maxilla for histological analysis. Results: This study showed a greater calcium marking in normotensive animals treated with losartan in relation to the other groups. Laser confocal microscopy parameters showed higher values of bone volume formed, mineralized surface, active surface of mineralization and bone formation rate in normotensive animals treated with losartan. However, a smaller mineralized surface was observed in all hypertensive animals. Conclusion: Losartan can improve bone mineralization parameters under normal physiological conditions, but the same anabolic effect does not occur under hypertension.

Cryotherapy: biochemical alterations involved in reduction of damage induced by exhaustive exercise

When exercises are done in intense or exhaustive modes, several acute biochemical mechanisms are triggered. The use of cryotherapy as cold-water immersion is largely used to accelerate the process of muscular recovery based on its anti-inflammatory and analgesic properties. The present study aimed to study the biochemical effects of cold-water immersion treatment in mice submitted to exercise-induced exhaustion. Swiss albino mice were divided into 4 treatment groups: control, cold-water immersion (CWI), swimming exhaustive protocol (SEP), and SEP+CWI. Treatment groups were subdivided into times of analysis: 0, 1, 3, and 5 days. Exhaustion groups were submitted to one SEP session, and the CWI groups submitted to one immersion session (12 min at 12°C) every 24 h. Reactive species production, inflammatory, cell viability, and antioxidant status were assessed. The SEP+CWI group showed a decrease in inflammatory damage biomarkers, and reactive species production, and presented increased cell viability compared to the SEP group. Furthermore, CWI increased acetylcholinesterase activity in the first two sessions. The present study showed that CWI was an effective treatment after exercise-induced muscle damage. It enhanced anti-inflammatory response, decreased reactive species production, increased cell viability, and promoted redox balance, which could decrease the time for the recovery process.

Endoscopic repair of cerebrospinal fluid rhinorrhea / Reparo endoscópico de fístulas liquóricas nasais

Abstract Introduction: Nasal liquorrhea indicates a cerebrospinal fluid fistula, an open communication between the intracranial cerebrospinal fluid and the nasal cavity. It can be traumatic and spontaneous. Objective: The aim of this study was to assess the outcome of endoscopic repair of cerebrospinal fluid fistula using fluorescein. Methods: This retrospective study included 30 patients of both sexes, with a mean age of 48.7 years, treated in the period from 2007 to 2015. All patients underwent lumbar administration of 5% sodium fluorescein solution preoperatively. Fistula was closed using three-layer graft and fibrin glue. Results: Cerebrospinal fluid fistulas were commonly located in the ethmoid (37%) and sphenoid sinus (33%). Most patients presented with traumatic cerebrospinal fluid fistulas (2/3 of patients). The reported success rate for the first repair attempt was 97%. Complications occurred in three patients: one patient presented with acute hydrocephalus, one with reversible encephalopathy syndrome on the fifth postoperative day with bilateral loss of vision, and one patient was diagnosed with hydrocephalus two years after the repair of cerebrospinal fluid fistula. Conclusion: Endoscopic diagnosis and repair of cerebrospinal fluid fistulas using fluorescein intrathecally has high success rate and low complication rate.

Resumo Introdução: A liquorreia nasal indica uma fístula liquórica (FL), uma comunicação aberta entre o líquido cerebrospinal intracraniano e a cavidade nasal. Pode ser traumática e espontânea. Objetivo: Avaliar o desfecho do reparo endoscópico da fístula liquórica nasal com o uso de fluoresceína. Método: Este estudo retrospectivo incluiu 30 pacientes de ambos os sexos, com idade média de 48,7 anos, tratados de 2007 a 2015. Todos os pacientes foram submetidos à administração lombar de solução de fluoresceína de sódio a 5% no pré-operatório. A fístula foi fechada com enxerto de três camadas e cola de fibrina. Resultados: As fístulas de líquido cerebrospinal foram comumente localizadas no seio etmoidal (37%) e esfenoidal (33%). A maioria dos pacientes apresentou fístulas liquóricas traumáticas (2/3). A taxa de sucesso relatada para a primeira tentativa de reparo foi de 97%. Complicações ocorreram em três pacientes: um apresentou hidrocefalia aguda; um, síndrome reversível de encefalopatia no quinto dia de pós-operatório com perda bilateral da visão; e um foi diagnosticado com hidrocefalia dois anos após o reparo de fístula liquórica. Conclusão: O diagnóstico endoscópico e o reparo de fístulas liquóricas nasais com uso de fluoresceína intratecal têm alta taxa de sucesso e baixo índice de complicações.

Obstrução congênita de vias lacrimais: série de casos de sondagem e intubação / Congenital lacrimal obstruction: case series of probing and intubation

RESUMO Objetivo: Avaliar a relação entre a idade da intervenção cirúrgica em pacientes portadores de obstrução congênita das vias lacrimais e o sucesso obtido pelos mesmos. Métodos: Foram estudados 94 prontuários de pacientes com diagnóstico de obstrução congênita de vias lacrimais, assistidos no setor de oculoplástica do Hospital de Olhos do Paraná, no período de janeiro de 2006 a dezembro de 2012. Para diagnóstico e inclusão na pesquisa foram realizados a anamnese, exames oftalmológico geral e específico utilizando o Teste de Milder e o Teste de Obtenção de Fluoresceína na Orofaringe. Para determinar o tipo de intervenção (sondagem ou intubação) foram utilizados os critérios do serviço. Na análise estatística foram utilizados o teste de qui-quadrado e o teste t de Student e adotado o nível de significância de 5%. Resultados: Quando comparados os resultados por idade, independente do tratamento, não houve associação significativa (p=0,223) entre a taxa de melhora e a idade. Conclusão: No presente estudo não houve diferença significativa entre a intervenção cirúrgica e as faixas etárias abordadas.

ABSTRACT Objective: To evaluate the relationship between age at surgery in patients with congenital lacrimal obstruction and success achieved by them. Methods: We studied 94 medical records of patients diagnosed with congenital lacrimal obstructions, assisted at the oculoplastic sector of the Hospital de Olhos do Paraná, in the period january 2006 to december 2012. For diagnosis and inclusion in the study, the clinical history were made, overall and specific ophthalmologic examination, with the tests: Milder and Observation Test of Fluorescein in the Oropharynx. To determine the type of intervention (probing or intubation) service criteria were used. In the statistical analysis, the chi-square and Student’s t test were used. Significance level of 5 % was adopted. Results: When comparing the results by age, regardless of treatment, there was no significant association (p = 0.223) between the rate of improvement and age. Conclusions: In the present study shows, there is no significant difference between the surgical interventions within the age groups addressed.

A method for combining Fluoro-Jade B staining and immunofluorescent staining / 南方医科大学学报

<p><b>OBJECTIVE</b>To explore a method for combining Fluoro-Jade B (FJB) staining with immunofluorescent staining in rats with focal cortical infarction.</p><p><b>METHOD</b>Permanent distal middle cerebral artery occlusion (dMCAO) was induced in rats by electrocoagulation. The rat models were randomized into two groups, and frozen sections of the brain tissues from each group were stained with FJB followed by immunofluorescent staining or in the reverse order.</p><p><b>RESULTS</b>FJB staining followed by immunofluorescence staining clearly visualized both FJB-positive and immunofluorescence-positive cells in the frozen sections, but the staining protocol in the reverse sequence failed to clearly show the immunofluorescence-positive cells.</p><p><b>CONCLUSION</b>FJB staining prior to immunofluorescence staining does not affect the staining effect of protein immunofluorescent staining and better visualizes the positive cells.</p>

Effects of intracellular Porphyromonas gingivalis on proliferation and osteogenic differentiation of human periodontal ligament cells in vitro / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the ability of Porphyromonas gingivalis to invade human periodontal ligament cells (hPDLCs) and the effect of intracellular P. gingivalis on cell proliferation and osteogenic differentiation in vitro.</p><p><b>METHODS</b>The invasion ability of P. gingivalis in hPDLCs was tested using an antibiotic protection assay at the multiplicity of infection (MOI) of 10 and 100. The proliferation of the infected cells was detected using a CFDA-SE kit, and the cells were sorted by fluorescence-activated cell sorting (FACS) followed by alizarin red staining for detecting mineralization nodules deposition; real-time PCR was used to examine the expression of Runx2 mRNA in the cells.</p><p><b>RESULTS</b>P. gingivalis actively invaded hPDLCs, and the internalized P. gingivalis was able to resist antibiotic treatment. The cells infected by P. gingivalis exhibited no significant suppression of cell proliferation, but showed significantly lowered capacity for osteogenic differentiation, down-regulated RUNX2 mRNA expression, and reduced mineral deposition.</p><p><b>CONCLUSION</b>Intracellular P. gingivalis does not significantly affect the proliferation of hPDLCs but inhibits osteogenic differentiation of the cells.</p>

Mitochondrial division inhibitor Mdivi-1 ameliorates angiotensin II-induced endothelial dysfunction / 生理学报

Mitochondrial fission can occur via activation of dynamin-related protein 1 (Drp1), which participates in the mitochondrial membrane scission process. The present study was designed to investigate the effect of angiotensin II (AngII) on mitochondrial fission and fusion in human umbilical vascular endothelial cells (HUVECs). And we further inquire into whether Mdivi-1, a newly identified pharmacological inhibitor of Drp1, can prevent endothelial dysfunction induced by AngII. The HUVECs were treated with AngII alone or in combination with Mdivi-1. Western blot was used to detect protein expressions of Drp1, endothelial nitric oxide synthase (eNOS) and apoptosis-related enzymes. MitoTracker Red and JC-1 dye were used to detect mitochondrial morphology and membrane potential, respectively. DCFH-DA probe was used to access intracellular reactive oxygen species (ROS) generation. Transwell assay was used to evaluate cell migration. Annexin V/PI staining was used to assess cellular apoptosis. The results showed that, in cultured HUVECs, AngII (1 × 10mol/L, 12 h) treatment significantly upregulated the expression of Drp1 followed by increased apoptosis and decreased eNOS expression. The treatment of AngII resulted in a change in mitochondrial morphology from elongated to uniformly punctate organelles, which was accompanied by decreased mitochondrial membrane potential. Furthermore, Mdivi-1 significantly protected against AngII-induced endothelial dysfunction, as shown by increased mitochondrial membrane potential and eNOS expression, reduced ROS level, decreased apoptosis and migration ability. Taking together, our data suggest that inhibition of Drp1 with Mdivi-1 can restore AngII-induced endothelial dysfunction.

Injurious effect of zinc deficiency on cardiomyocytes / 生理学报

The aim of the present study was to investigate the effect of zinc deficiency on cardiomyocyte survival and the underlying mechanisms. Simulated zinc deficiency model was developed in H9c2 cardiac cells with zinc chelator N, N, N', N'-tetrakis (2-pyridylmethyl) ethylenediamine (TPEN). MTT assay was used to evaluate cell viability. Morphological changes of the cells were observed by optical microscope. Lacate dehydrogenase (LDH) levels of the cells were determined with LDH assay kit. Mitochondrial membrane potential (ΔΨ) was measured with confocal microscope using JC-1 dye. Intracellular reactive oxygen species (ROS) levels were determined by DCFH-DA staining. PD98059 (an inhibitor of ERK), SNAP, which can activate ERK, and the ROS scavenger, MPG, were respectively used to investigate mechanism of signal transduction. The phosphorylation of ERK was detected by Western blot. The results showed that TPEN significantly induced the cell morphological damage and the loss of ΔΨ, increased LDH leakage, and promoted ROS generation. In the H9c2 cells, TPEN significantly inhibited ERK phosphorylation and decreased cell viability, which was potentiated by PD98059, whereas both SNAP and MPG reversed the inhibitory effects of TPEN. These data suggest that zinc deficiency leads to the injury in H9c2 cardiac cells through down-regulating ERK pathway. Increased intracellular ROS may account for the effect of zinc deficiency.

Fluorescein Diacetate Microplate Assay in Cell Viability Detection / 中国医学科学院学报

Objective To investigate the application of the fluorescein diacetate (FDA) microplate assay in cell viability detection. Methods Cells were seeded in a 96-well culture plate until detection. After incubated with FDA,the plate was detected by fluorescence microplate analyzer. The effects of FDA incubation duration,concentration,and other factors on the assay's accuracy and stability were assessed. We also compared the results of FDA with methyl thiazolyl(MTT) in terms of cell numbers and HOinjury. Results Within 0-30 minutes,the fluorescence-cell number coefficient of FDA assay increased with duration and reached 0.99 in 27-30 minutes. The optimum concentration of final FDA in this study was 10-30 μg/ml. On cell viability detection,the result of FDA method was equivalent to MTT method. As to HOinjury assay,the sensitivity of FDA method was superior to MTT on the higher concentration HOtreatment due to a relative shorter duration for detection. Conclusion As a stable and reliable method,FDA is feasible for cell variability detection under varied conditions.

Chemical and cytological analysis of cerebral spinal fluid after intrathecal injection of hypodense fluorescein / Análise química e citológica do líquor após injeção intratecal de solução hipodensa de fluoresceína

ABSTRACT INTRODUCTION: Intrathecal fluorescein has been effective for topographic diagnosis of rhinoliquorrhea. Nonetheless, there are no reports on the study of cerebral spinal fluid (CSF) after use of intrathecal fluorescein. OBJECTIVE: A prospective study attempting to evaluate CSF through chemical and cytological analysis, after injection of fluorescein. METHODS: Prospective analysis of 24 samples of CSF after intrathecal injection of fluorescein for topographic diagnosis of CSF fistulae, collected at the time of puncture and after 24 and 48 h, divided by cellularity: Group 1, up to five cells, and Group 2, with more than five cells. RESULTS: The yellow-greenish color of CSF remained after 48 h in 36%, evidencing permanence of fluorescein. No changes in protein and glucose levels were observed between 0-24 h and 0-48 h. In group 2, an increase in cell count was observed between 24 h and 48 h (p = 0.019). In both groups, there was an increase of neutrophils between 0 and 48 h (p = 0.048) and a decrease between 24 and 48 h (p = 0.05). CONCLUSION: Intrathecal fluorescein provoked discreet meningeal reactions, such as an increase of cells between 24 and 48 h and an increase of neutrophils at 24 h, with a subsequent decrease at 48 h with no correlation with symptomatology.

RESUMO Introdução: A fluoresceína intratecal tem sido efetiva no diagnóstico topográfico da rinoliquorréia. Entretanto, não há estudos no líquor após o uso de fluoresceína intratecal. Objetivo: Estudo prospectivo visando avaliar o líquor, através de análise química e citológica, após injeção de fluoresceína. Método: Análise prospectiva de 24 punções após injeção intratecal de fluoresceína para diagnóstico topográfico de fístula liquórica, coletado no momento da punção, 24 e 48 horas, divididos pela celularidade: grupo 1, com até 5 células e grupo 2 com mais de 5 células. Resultado: A coloração amarelo-esverdeada do líquor permaneceu após 48 horas em 36%, evidenciando permanência de fluoresceína. Observou-se ausência de mudanças no nível de proteína e glicose entre 0-24 horas e 0-48 horas. No grupo 2, um aumento na contagem celular foi observado entre 24 e 48 horas (p = 0,019). No dois grupos juntos, observou-se um aumento de neutrófilos entre 0 e 48 horas (p = 0,048) e uma diminuição entre 24 e 28 horas (p = 0,05). Conclusão: Fluoresceína intratecal provocou discretas reações meníngeas, como o aumento de células entre 24 e 48 horas e aumento dos dos neutrófilos em 24 horas com uma subsequente dimi nuição em 48 horas sem correlação com sintomas.

Resección microquirúrgica de glioblastoma guiada con fluoresceína intraoperatoria: evaluación retrospectiva / Microsurgical resection of glioblastoma guided with intraoperative fluorescein: a retrospective evaluation

Evaluar la influencia del uso de fluoresceína sódica (FLS-Na) en la cirugía del glioblastoma (GB) sobre el grado de resección tumoral y la supervivencia en pacientes atendidos en el Instituto Nacional de Enfermedades Neoplásicas. Materiales y métodos. Se revisó un total de 238 casos de GB atendidos entre los años 2008 y 2013 y se seleccionó 150 casos de GB sometidos a resección quirúrgica, con información clínico-patológica y seguimiento adecuado. Resultados. La media de edad fue 51 años, el 58,7% de casos presento Karnofsky de al menos 90. Se administró FLS-Na en 80 casos (53,3%) y se obtuvo una resección subtotal y total en 69 (46%) y 81 (54%) de los casos, respectivamente. El grupo que recibió FLS-Na obtuvo mayores tasas de resección total que el grupo operado solo con luz blanca (77,5 vs 27,1%, p<0,001). La mediana de sobrevida global (SG) fue mayor en el grupo sometido a resección total que a subtotal (17 vs 7 meses, p<0,001). La mediana de SG en los que recibieron FLS-Na fue mayor que en los que no la recibieron (15,0 vs 8 meses, p=0,003). Otros factores que afectaron la SG fueron la edad (p=0,002), el Karnofsky (p=0,052) y la administración de radioterapia (p=0,016) y quimioterapia (p=0,011). Conclusiones. La técnica microquirúrgica con administración de FLS-Na se asoció con un aumento en la tasa de resecciones totales y de supervivencia...

To evaluate the influence of the use of sodium fluorescein (FLS-Na) in surgery of glioblastoma (GB) on the degree of tumor resection and survival in patients treated at the National Institute of Neoplastic Diseases. Materials and methods. A total of 238 cases of GB treated between 2008 and 2013 were reviewed and 150 cases of GB who underwent surgical resection with clinicopathological information and adequate follow-up were selected. Results. The mean age was 51 years, 58.7% of the cases presented a Karnofsky score of at least 90. FLS-Na was administered in 80 cases (53.3%) and a subtotal and total resection was obtained in 69 (46%) and 81 (54%) cases, respectively. The group that received FLS-Na obtained higher rates of total resection than the group operated with white light alone (77.5 vs 27.1%, p<0.001). The median overall survival (OS) was higher in the group subject to total compared to subtotal resection (17 vs 7 months, p<0.001). The median OS in those who received FLS-Na was higher than in those who did not (15.0 vs 8 months, p=0.003). Other factors affecting OS were age (p=0.002), the Karnofsky score (p=0.052) and radiation therapy (p=0.016) and chemotherapy (p=0.011). Conclusions. The microsurgical technique with administration of FLS-Na was associated with an increase in the rate of total resection and survival...

Neuroprotective effects of paeonol in a cell model of Parkinson disease / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To investigate the effects of paeonol on neuron cell model of Parkinson disease (PD).</p><p><b>METHODS</b>The cell model of Parkinson disease was induced by treatment of 1-Methyl-4-phenylpyridinium (MPP+) in PC12 cells, the PD model cells were treated with 1 μmol/L, 3 μmol/L or 9 μmol/L paeonol for 24h, respectively. Cell viability and LDH leakage were detected by MTT and lactate dehydrogenase (LDH) assay; the apoptosis of PC12 cells was assessed by Hoechst 33258 staining and flow cytometry; reactive oxygen species (ROS) production was detected by DCFH-DA method; and the ratio of Bax/Bcl-2 and activation of caspase-3 were determined by Western blotting.</p><p><b>RESULTS</b>MPP+ treatment significantly reduced cell viability, increased LDH leakage, enhanced the proportion of apoptotic cells and ROS production. In addition, MPP+ treatment dramatically increased the Bax/Bcl-2 ratio, and the activation of caspase-3. Compared to PD model group, paeonol treatment significantly enhanced cell viability, decreased LDH leakage, inhibited the proportion of apoptotic cells and ROS production, reduced the Bax/Bcl-2 ratio and the activated caspase-3 protein.</p><p><b>CONCLUSION</b>Paeonol can prevent PC12 cells from apoptosis induced by MPP+, and the mechanism may be associated with the down-regulation of ROS production, Bax/Bcl-2 ratio and Caspase-3 activation.</p>

Involvement of oxidative stress in embelin-induced cell death in leukemia HL-60 cells / 中华血液学杂志

<p><b>OBJECTIVE</b>To evaluate the effects of Embelin on HL-60 cells by the impact of oxidative stress on DNA double-strain breaks (DSBs).</p><p><b>METHODS</b>HL-60 cells were treated with Embelin in different concentration (3, 10, 30, 100, and 300 μg/ml) for 24 h, and inhibitory effects was examined by CCK-8 assay. Reactive oxygen species (ROS) levels were evaluated by flow cytometry using DCFH-DA. Comet assay was used to detect the extent of DSBs.</p><p><b>RESULTS</b>Embelin inhibited proliferation of HL-60 cells in a dose-dependent manner. At the concentration of 10, 30, 100, and 300 μg/ml, the inhibition rate was (12.74 ± 2.27)%, (23.49 ± 1.96)%, (30.30±1.89)%, and (57.55 ± 3.59)% (P<0.05). Embelin also lead to high level of intracellular ROS and deterioration of DNA damage (P<0.05). When HL-60 cells were pretreated with ROS scavenger N-acetyl-l-cysteine (NAC) for 2 h and then treated with 300 μg/ml Embelin for 24 h, the intracellular ROS level declined and DSBs relieved (P<0.05). Meanwhile, embelin-induced cell viability significantly declined to (32.75 ± 2.70)% (P<0.05).</p><p><b>CONCLUSION</b>Embelin induced the death of HL-60 cells by increasing the generation of intracellular oxidation and the oxidative stress, which drived the damage of DNA double-strand.</p>

Correlation between Fluorescein Angiographic Findings and Visual Acuity in Behcet Retinal Vasculitis

PURPOSE: To identify significant fluorescein angiographic (FA) characteristics associated with visual acuity (VA) in Behcet retinal vasculitis. MATERIALS AND METHODS: Retrospective review of 86 eyes of 48 patients (age: 35.6+/-10.2 years) with Behcet retinal vasculitis were performed. VA and FA findings as well as correlation between them were assessed. RESULTS: The mean initial VA of eyes with posterior pole-involved vasculitis (63 eyes; 73.3%) was significantly worse than that of those with peripheral vasculitis (23 eye; 26.7%) (logarithm of the minimum angle of resolution VA: 0.554+/-0.572 vs. 0.078+/-0.148; p<0.0001). Subgroup analysis revealed a more severe and diffuse pattern of vascular leakage in posterior pole-involved vasculitis compared to peripheral vasculitis (p<0.0001). Retinal vascular leakage (beta=0.345; p<0.0001), optic disc hyperfluorescence (beta=0.147; p=0.032), and macular leakage (beta=0.107; p=0.047) were significantly associated with worse initial VA. During the follow up (mean: 33.3+/-17.9 months), the change of leakage showed no significant correlation with change of VA in posterior pole-involved vasculitis (tau=0.199, p=0.092). CONCLUSION: Posterior pole involvement, the degree of retinal vascular leakage, optic disc hyperfluorescence, and macular leakage are significantly associated with VA in Behcet retinal vasculitis.

In vivo Cell CFSE Fluorescence Negative Staining for Detection of Super Paramagnetic Iron Oxide Nanoparticles Phagocytosed by Mouse Mononuclear Macrophage Leukemia Cells-RAW264.7 / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To explore the feasibility and fluorescence characteristics of CFSE negative staining for in vivo cell imaging of super paramagnetic iron oxide nanoparticles (SPIO) phagocytosed by mouse mononuclear macrophage leukemia cells-RAW264.7.</p><p><b>METHODS</b>After labeled with SPIO, the RAW264.7 macrophages were stained with Prussian blue stain and CFSE fluorescence negative stain step by step. Furthermore, trypan blue staining was used to evaluate cell viability of cells which stained with CFSE. At last, laser scanning confocal microscope was used to measure SPIO in cells through CFSE fluorescence negative stain method.</p><p><b>RESULTS</b>SPIO within RAW264.7 macrophages showed blue in Prussian's blue staining, while showed negative area in CFSE negative staining. Good consistencies between Prussian's blue staining and CFSE negative staining were observed. In addition, RAW264.7 macrophages showed high viability after SPIO/CFSE dual-labeled method, proved by typan stain.</p><p><b>CONCLUSION</b>The CFSE fluorescence negative staining may be used for detecting SPIO that phagocytosed by RAW264.7 macrophages and it is showed good consistency that confirmed one another when compared to classic Prussian' blue staining.</p>

Síndrome de Fraser: relato de caso nas vias lacrimais / Fraser Syndrome: case report in lacrimal system

A síndrome de Fraser é uma condição sistêmica caracterizada por criptoftalmo, sindactilia e anomalia da genitália, podendo se associar com alterações dos rins, do ouvido, do nariz, da laringe e do esqueleto. O criptoftalmo pode representar um achado isolado, representado por herança autossômica dominante, associado a outras anomalias congênitas, relatado como herança autossômica recessiva. Criança do sexo feminino, 9 meses, avaliada no ambulatório de vias lacrimais da Universidade Federal de São Paulo. Filha de pais consanguíneos. Ao exame, foram observados criptoftalmo total à esquerda, epífora em olho direito associada à secreção mucopurulenta, nariz em sela, implantação baixa das orelhas, malformação de conduto auditivo, aumento de grandes lábios e sindactilia de mãos e pés. A tomografia de crânio evidenciou braquicefalia ausência de septo pelúcido, proeminência dos ventrículos laterais, importante falha óssea na calota craniana, presença de afilamento do manto tecidual cerebral, fossa posterior pequena, desorganização do segmento anterior, afacia e descolamento total da retina.

Fraser syndrome is a systemic condition characterized by cryptophthalmos, syndactyly and abnormal genitalia, which may be associated with urinary tract, ear, nose, larynx and skeletal abnormalities. Cryptophthalmos can be an isolated finding (that has been reported as an autosomal dominant trait) or associated with other congenital anomalies (reported as an autosomal recessive disorder). Child, female, nine month of life, evaluated in the lacrimal setor of Federal University of São Paulo. Child of consanguineous parents. Her physical examination showed total unilateral cryptophthalmos (left side), epiphora (right side) with mucopurulent discharge, depressed nasal bridge, low set ears, atresia of the external auditory canal, prominent labia majora and syndactyly of the fingers and toes. Ocular ultrasonography showed brachycephaly, absence of septu pellucidum prominence of the lateral ventricles, a major bone defect in the skull, the presence of thinning of the mantle tissue of the brain,a reduced anterior-posterior ocular diameter, anterior segment disorganization, absence of the lens and total retinal detachment in the left eye.

Modification by wheat germ agglutinin delays the ocular elimination of liposome / 药学学报

The purpose of this study is to explore the feasibility of wheat germ agglutinin (WGA) modified liposome as a vehicle for ophthalmic administration. Liposome loaded with 5-carboxyfluorescein (FAM) was prepared by lipid film hydration method. WGA was thiolated and then conjugated to the surface of the liposome via polyethylene glycol linker to constitute the WGA-modified and FAM-loaded liposome (WGA-LS/FAM). The amount of thiol groups on each WGA molecule was determined, and the bioactivity of WGA was estimated after it was modified to the surface of liposome. The physical and chemical features of the WGA-modified liposome were characterized and the ocular bioadhesive performance was evaluated in rats. The result showed that each thiolated WGA molecule was conjugated with 1.32 thiol groups. WGA-LS/FAM had a mean size of (97.40 +/- 1.39) nm, with a polydispersity index of 0.23 +/- 0.01. The entrapment efficacy of FAM was about (2.95 +/- 0.21)%, and only 4% of FAM leaked out of the liposome in 24 h. Erythrocyte agglutination test indicated that after modification WGA preserved the binding activity to glycoprotein. The in vivo ocular elimination of WGA-LS/FAM fitted first-order kinetics, and the elimination rate was significantly slower than that of the unmodified liposome, demonstrating WGA-modified liposome is bioadhesive and suitable for ophthalmic administration.

Análisis comparativo de marcadores de lesión en modelos de isquemia cerebral focal y global en ratas / Injury markers in two models of cerebral ischemia

Introducción. Los indicadores espacio-temporales de lesión son esenciales en el estudio neuropatológico y terapéutico de la isquemia cerebral. Objetivo. Optimizar la técnica de dos modelos de isquemia cerebral (focal y global) y hacer un análisis comparativo de la progresión del daño cerebral, mediante marcadores de neurodegeneración. Materiales y métodos. Se sometieron ratas Wistar a oclusión temporal de la arteria cerebral media o a oclusión de cuatro vasos, y se evaluaron comparativamente el tiempo quirúrgico, la tasa de supervivencia y la recuperación neurológica. Se utilizó trifenilo de tetrazolio para establecer la distribución del infarto y tinción con Fluoro - Jade B ® como marcador de neurodegeneración. La inmunorreacción de la astroglía se evaluó con el anticuerpo contra la proteína acídica fibrilar de la glía ( Glial Fibrillary Acidic Protein, GFAP) y el anticuerpo AT-8 contra la proteína tau hiperfosforilada, 24, 48 y 72 horas después de la isquemia. Resultados. Los modelos de isquemia utilizados requirieron menor tiempo quirúrgico y hubo menor riesgo de muerte, respecto a estudios previos. En el modelo focal, las células positivas con Fluoro - Jade B ® y los astrocitos reactivos, se evidenciaron en corteza e hipocampo a las 24 horas después de la isquemia. En el modelo global, se observó tinción Fluoro - Jade B ® positiva a las 24 horas, aumentando significativamente la reacción de la GFAP a las 72 horas en corteza y a las 48 horas en el hipocampo. La reacción contra la proteína tau hiperfosforilada aumentó progresivamente y fue máxima a las 72 horas en ambos modelos. Conclusiones. Los dos modelos de isquemia cerebral, oclusión temporal de la arteria cerebral media y oclusión de cuatro vasos, fueron optimizados. En estos modelos, los marcadores la tinción Fluoro - Jade B ® y la GFAP permitieron detectar procesos de neurodegeneración 24 horas después de la isquemia, en tanto el marcador de proteína tau hiperfosforilada (AT-8) incrementó progresivamente su reacción hasta las 72 horas, lo cual sugiere la propagación de la excitotoxicidad y la alteración de enzimas implicadas en la fosforilación de proteínas del citoesqueleto.

Introduction: Spatio-temporal indicators of injury are essential for the study of neuropathological processes and for developing therapeutic approaches for stroke. Objective: This study sought to optimize the techniques of two cerebral ischemia models (focal and global) and to comparatively evaluate the progression of brain damage by analyzing markers of neurodegeneration. Materials and methods: Wistar rats were subjected to temporary occlusion of the middle cerebral artery (t-MCAO) or four-vessel occlusion (4-VO), and surgical time, survival rate and neurological recovery were comparatively evaluated. Triphenyl tetrazolium was used to determine the distribution of the infarction, and Fluoro-Jade B was used as a marker of neurodegeneration. Astroglial immunoreactivity was assessed with an anti-glial fibrillary acidic protein (GFAP) antibody, and an anti-AT-8 antibody was used to detect hyperphosphorylated tau protein at 24, 48 and 72 hours post-ischemia. Results: The cerebral ischemia models employed (t-MCAO and 4-VO) required less surgical time and presented less of a death risk compared to those in previous studies. In the focal model, Fluoro-Jadepositive cells and reactive astrocytes were observed in the cerebral cortex and the hippocampus at 24 hours post-ischemia. In the global model, we observed Fluoro-Jade-positive cells at 24 hours, and a significant increase in the reactivity of GFAP was observed at 72 hours in the cortex and at 48 hours in the hippocampus. The immunoreactivity of hyperphosphorylated tau protein increased progressively, reaching a maximum at 72 hours post-ischemia in both models. Conclusions: These results suggest that in the t-MCAO and 4-VO ischemia models, the expression of Fluoro-Jade and GFAP indicates early neurodegeneration at 24 hours post-insult. In contrast, the immunoreactivity of the hyperphosphorylated tau protein marker (AT-8) progressively increases until 72 hours post-insult, which suggests that the progression of excitotoxicity and alteration of enzymes involves the phosphorylation of cytoskeletal proteins.

Systemic administration of low dosage of tetanus toxin decreases cell proliferation and neuroblast differentiation in the mouse hippocampal dentate gyrus / 한국실험동물학회지

In the present study, we investigated the effect of Tetaus toxin (TeT) on cell proliferation and neuroblast differentiation using specific markers: 5-bromo-2-deoxyuridine (BrdU) as an exogenous marker for cell proliferation, Ki-67 as an endogenous marker for cell proliferation and doublecortin (DCX) as a marker for neuroblasts in the mouse hippocampal dentate gyrus (DG) after TeT treatment. Mice were intraperitoneally administered 2.5 and 10 ng/kg TeT and sacrificed 15 days after the treatment. In both the TeT-treated groups, no neuronal death occurred in any layers of the DG using neuronal nuclei (NeuN, a neuron nuclei maker) and Fluoro-Jade B (F-J B, a high-affinity fluorescent marker for the localization of neuronal degeneration). In addition, no significant change in glial activation in both the 2.5 and 10 ng/kg TeT-treated-groups was found by GFAP (a marker for astrocytes) and Iba-1 (a marker for microglia) immunohistochemistry. However, in the 2.5 ng/kg TeT-treated-group, the mean number of BrdU, Ki-67 and DCX immunoreactive cells, respectively, were apparently decreased compared to the control group, and the mean number of each in the 10 ng/kg TeT-treated-group was much more decreased. In addition, processes of DCX-immunoreactive cells, which projected into the molecular layer, were short compared to those in the control group. In brief, our present results show that low dosage (10 ng/kg) TeT treatment apparently decreased cell proliferation and neuroblast differentiation in the mouse hippocampal DG without distinct gliosis as well as any loss of adult neurons.