Prediction of protein subcellular localization based on multilayer sparse coding / 生物工程学报

In order to provide a theoretical basis for better understanding the function and properties of proteins, we proposed a simple and effective feature extraction method for protein sequences to determine the subcellular localization of proteins. First, we introduced sparse coding combined with the information of amino acid composition to extract the feature values of protein sequences. Then the multilayer pooling integration was performed according to different sizes of dictionaries. Finally, the extracted feature values were sent into the support vector machine to test the effectiveness of our model. The success rates in data set ZD98, CH317 and Gram1253 were 95.9%, 93.4% and 94.7%, respectively as verified by the Jackknife test. Experiments showed that our method based on multilayer sparse coding can remarkably improve the accuracy of the prediction of protein subcellular localization.

Effects of Hedan Tablet () on lipid profile, proprotein convertase subtilisin/kexin type 9 and high-density lipoprotein subfractions in patients with hyperlipidemia: A primary study / 中国结合医学杂志

<p><b>OBJECTIVE</b>To investigate the effects of Hedan Tablet () on serum lipid profile, proprotein convertase subtilisin/kexin type 9 (PSCK9) and high-density lipoprotein (HDL) subfractions in patients with hyperlipidemia.</p><p><b>METHODS</b>Thirty-seven patients with hyperlipidemia were randomized to treatment with Hedan Tablet 4.38 g/day as Hedan group (18 cases) or placebo (19 cases) as control group for 8 weeks. The lipid profile, PCSK9 and HDL subfractions were determined at day 0 and week 8 in both groups respectively.</p><p><b>RESULTS</b>Hedan treatment for 8 weeks mildly decreased serum low-density lipoprotein cholesterol (LDL-C) levels, while no changes were found in total cholesterol (TC), triglycerides (TG) and PCSK9 concentrations. Furthermore, Hedan treatment increased the concentration of large high-density lipoprotein cholesterol (HDL-C) and the percentage of large HDL subfraction, while decreased the concentration of small HDL-C and the percentage of small HDL subfraction without changing serum HDL-C levels in patients with hyperlipidemia.</p><p><b>CONCLUSION</b>Hedan treatment of 4.38 g per day for 8 weeks could confer a favorable effects on serum LDL-C concentration as well as HDL subfractions.</p>

Scientific use of the finite element method in Orthodontics

INTRODUCTION: The finite element method (FEM) is an engineering resource applied to calculate the stress and deformation of complex structures, and has been widely used in orthodontic research. With the advantage of being a non-invasive and accurate method that provides quantitative and detailed data on the physiological reactions possible to occur in tissues, applying the FEM can anticipate the visualization of these tissue responses through the observation of areas of stress created from applied orthodontic mechanics. OBJECTIVE: This article aims at reviewing and discussing the stages of the finite element method application and its applicability in Orthodontics. RESULTS: FEM is able to evaluate the stress distribution at the interface between periodontal ligament and alveolar bone, and the shifting trend in various types of tooth movement when using different types of orthodontic devices. Therefore, it is necessary to know specific software for this purpose. CONCLUSIONS: FEM is an important experimental method to answer questions about tooth movement, overcoming the disadvantages of other experimental methods. .

INTRODUÇÃO: o Método de Elementos Finitos (MEF) é um recurso da Engenharia empregado para calcular o estresse e a deformação de estruturas complexas, e tem sido amplamente utilizado nas pesquisas em Ortodontia. Apresenta a vantagem de ser um método não-invasivo e preciso, que fornece dados quantitativos e detalhados acerca das reações fisiológicas que podem ocorrer nos tecidos. OBJETIVO: esse artigo pretende realizar uma revisão da literatura sobre as etapas para realização do Método de Elementos Finitos, bem como de sua aplicabilidade na Ortodontia. RESULTADOS: o MEF é capaz de avaliar a distribuição do estresse na interface entre o ligamento periodontal e o osso alveolar, bem como a tendência de deslocamento em diversos tipos de movimentos dentários, quando utilizados diferentes tipos de aparelhos. Para tanto, é necessário conhecimento de softwares específicos para esse fim. CONCLUSÕES: o MEF é um importante método experimental que pode esclarecer questionamentos acerca da movimentação dentária, superando as desvantagens de outros métodos experimentais. .

Adaptation and validation of the Caregiver Burden Inventory for use with caregivers of elderly individuals / Adaptação e validação do Inventário de Sobrecarga do Cuidador para uso em cuidadores de idosos / Adaptación y validación del Inventario de Sobrecarga del Cuidador para uso en cuidadores de ancianos

OBJECTIVE: to adapt and validate the Caregiver Burden Inventory for use with caregivers of older adults in Brazil. METHOD: methodological study involving initial translation, synthesis of translations, back translation, expert committee review, pre-testing, submission of the final version to the original authors, and assessment of the inventory's psychometric properties. The inventory assesses five dimensions of caregiver burden: time-dependence, developmental, physical, social and emotional dimensions. RESULTS: a total of 120 family caregivers took part in the study. All care-receivers were older adults dependent on assistance to perform activities of daily living, and lived in the central region of the city of Porto Alegre, RS, Brasil. Cronbach's alpha value for the inventory was 0.936, and the Pearson correlation coefficient for the relationship between the scores obtained on the Caregiver Burden Inventory and the Burden Interview was 0.814. The intraclass correlation coefficient was 0.941, and the value of Student's T-test comparing test and retest scores was 0.792. CONCLUSION: the instrument presented adequate reliability and the suitability of its items and factors was confirmed in this study. .

OBJETIVO: adaptar e validar o Inventário de Sobrecarga do Cuidador para uso em cuidadores de idosos no Brasil. MÉTODO: estudo metodológico envolvendo tradução inicial, síntese das traduções, retrotradução, revisão por comitê de especialistas, pré-teste, envio da versão final para apreciação dos autores da versão original, e avaliação de suas propriedades psicométricas. O inventário avalia cinco dimensões de sobrecarga do cuidador: tempo dependente, sobrecarga à vida pessoal, física, social e emocional. RESULTADOS: participaram do estudo 120 cuidadores familiares. Todos os indivíduos sob cuidado destes cuidadores eram idosos que dependiam de assistência para realizar atividades da vida diária e residiam na região central da cidade de Porto Alegre, RS, Brasil. O valor alfa de Cronbach encontrado para o inventário foi de 0,936 e o coeficiente de correlação Pearson para a relação entre os escores obtidos no Inventário de Sobrecarga do Cuidador e na escala Burden Interview foi de 0,814. O coeficiente de correlação intraclasse foi de 0,941 e o valor do teste t de Student que comparou os escores do teste e reteste foi de 0,792. CONCLUSÃO: o instrumento apresentou confiabilidade apropriada e a adequação de seus itens e domínios foi confirmada neste estudo. .

OBJETIVO: adaptar y validar el Inventario de Sobrecarga del Cuidador para uso en cuidadores de ancianos en Brasil. MÉTODO: estudio metodológico con traducción inicial, síntesis de las traducciones, retrotraducción, revisión por comité de especialistas, preprueba, envío de la traducción final para apreciación de los autores del instrumento original, y evaluación de sus propiedades psicométricas. El Inventario evalúa cinco dominios de sobrecarga del cuidador: tiempo dependiente, sobrecarga en la vida personal, física, social y emocional. RESULTADOS: participaron del estudio 120 cuidadores familiares. Todos los individuos bajo el cuidado de estos cuidadores eran ancianos que dependían de asistencia para realizar actividades de la vida diaria y residían en la región central de la ciudad de Porto Alegre, RS, Brasil. El valor Alfa de Cronbach encontrado para el Inventario fue 0,936 y el coeficiente de correlación de Pearson para la relación entre los puntajes obtenidos entre el Inventario de Sobrecarga del Cuidador y el Burden Interview fue 0,814. El coeficiente de correlación intraclase fue 0,941 y el valor de la prueba t de Student que comparó los puntajes de la prueba y reprueba fue de 0,792. CONCLUSIÓN: el instrumento presentó confiabilidad apropiada y la adecuación de sus ítems y dominios fue confirmada en este estudio. .

Cultural adaptation, content validity and inter-rater reliability of the "STAR Skin Tear Classification System" / Adaptação cultural, validade de conteúdo e confiabilidade interobservadores do "STAR Skin Tear Classification System" / Adaptación cultural, validez de contenido y confiabilidad interobservadores del "STAR Skin Tear Classification System"

AIMS: to perform the cultural adaptation of the STAR Skin Tear Classification System into the Portuguese language and to test the content validity and inter-rater reliability of the adapted version. METHODS: methodological study with a quantitative approach. The cultural adaptation was developed in three phases: translation, evaluation by a committee of judges and back-translation. The instrument was tested regarding content validity and inter-rater reliability. RESULTS: the adapted version obtained a regular level of concordance when it was applied by nurses using photographs of friction injuries. Regarding its application in clinical practice, the adapted version obtained a moderate and statistically significant level of concordance. CONCLUSION: the study tested the content validity and inter-rater reliability of the version adapted into the Portuguese language. Its inclusion in clinical practice will enable the correct identification of this type of injury, as well as the implementation of protocols for the prevention and treatment of friction injuries. .

OBJETIVOS: realizar a adaptação cultural do STAR Skin Tear Classification System, para a língua portuguesa e testar a validade de conteúdo e a confiabilidade interobservadores da versão adaptada. MÉTODOS: estudo metodológico com abordagem quantitativa. A adaptação cultural foi desenvolvida em três fases: tradução, avaliação por comitê de juízes e retrotradução. O instrumento foi testado quanto à validade de conteúdo e confiabilidade interobservadores. RESULTADOS: a versão adaptada obteve um nível regular de concordância quando aplicada por enfermeiros em fotografias de lesões por fricção. Quando aplicada na prática clínica, a versão adaptada obteve nível moderado e estatisticamente significativo de concordância. CONCLUSÃO: o estudo atestou a validade de conteúdo e a confiabilidade interobservadores da versão adaptada para a língua portuguesa. Sua inclusão na prática clínica possibilitará a correta identificação desse tipo de lesão, além da implementação de protocolos para a prevenção e tratamento das lesões por fricção. .

OBJETIVOS: realizar la adaptación cultural del STAR Skin Tear Classification System, para el idioma portugués y comprobar la validez de contenido y la confiabilidad interobservadores de la versión adaptada. MÉTODOS: estudio metodológico con abordaje cuantitativo. La adaptación cultural fue desarrollada en tres fases: traducción, evaluación por comité de jueces y retrotraducción. El instrumento fue comprobado en lo que se refiere a su validez de contenido y confiabilidad interobservadores. RESULTADOS: la versión adaptada obtuvo un nivel regular de concordancia cuando fue aplicada por enfermeros utilizando fotografías de lesiones por fricción. Cuando fue aplicado en la práctica clínica, la versión adaptada obtuvo un nivel moderado y estadísticamente significativo de concordancia. CONCLUSIÓN: el estudio comprobó la validez de contenido y la confiabilidad interobservadores de la versión adaptada para el idioma portugués. Su inclusión en la práctica clínica posibilitará la correcta identificación de ese tipo de lesión, además de la implementación de protocolos para la prevención y tratamiento de las lesiones por fricción. .

Mitochondrial ATPase activity and membrane fluidity changes in rat liver in response to intoxication with Buckthorn (Karwinskia humboldtiana)

BACKGROUND: Karwinskia humboldtiana (Kh) is a poisonous plant of the rhamnacea family. To elucidate some of the subcellular effects of Kh toxicity, membrane fluidity and ATPase activities as hydrolytic and as proton-pumping activity were assessed in rat liver submitochondrial particles. Rats were randomly assigned into control non-treated group and groups that received 1,1.5 and 2 g/Kg body weight of dry powder of Kh fruit, respectively. Rats were euthanized at day 1 and 7 after treatment. RESULTS: Rats under Kh treatment at all dose levels tested, does not developed any neurologic symptoms. However, we detected alterations in membrane fluidity and ATPase activity. Lower dose of Kh on day 1 after treatment induced higher mitochondrial membrane fluidity than control group. This change was strongly correlated with increased ATPase activity and pH gradient driven by ATP hydrolysis. On the other hand, membrane fluidity was hardly affected on day 7 after treatment with Kh. Surprisingly, the pH gradient driven by ATPase activity was significantly higher than controls despite an diminution of the hydrolytic activity of ATPase. CONCLUSIONS: The changes in ATPase activity and pH gradient driven by ATPase activity suggest an adaptive condition whereby the fluidity of the membrane is altered.

Cell accumulation and antileishmanial effect of exogenous and endogenous protoporphyrin IX after photodynamic treatment / Acumulación celular y efecto anti- Leishmania de la protoporfirina IX exógena y endógena después del tratamiento fotodinámico

Introduction: Photodynamic therapy (PDT) using 5-aminolevulinic acid-induced protoporphyrin IX (ALA-PpIX) constitutes an interesting alternative for cutaneous leishmaniasis treatment. Objective: To evaluate the production of PpIXbased on the administration of ALA and MAL and the effect of ALA-PDTat cellular level on non-infected and infected THP-1 cells using Leishmania ( Viannia ) panamensis or Leishmania ( Leishmania ) infantum (syn Leishmania chagasi ) parasites. Materials and methods: Protoporphyrin IX (PpIX) production and mitochondrial colocalization were evaluated by confocal microscopy. Cell toxicities were evaluated after treatment with the compounds, followed by light irradiation (597-752 nm) at 2.5 J/cm 2 fluency using a colorimetric MTT assay for THP-1 cells and a standard microscopic analysis of parasites. Results were expressed as compound concentration activity against 50% of cells or parasites (CC 50 or IC 50 ). Results: ALA or MAL induced an endogenous PpIX with a red fluorescence localized mainly in the mitochondria inside human cells. ALA and MAL-PDT induced a similar range of toxicities on THP-1 cells (CC 50 0.16±0.01mM and 0.33±0.019 mM, respectively) without any apparent inhibition of intracellular parasites in the infected cells as compared to untreated controls. Exogenous PpIX-PDT was toxic to THP-1 cells (CC 50 0.00032±0.00002 mM), L. (L.) infantum (IC 50 0.003±0.0001 mM) and L. (V.) panamensis (IC 50 0.024±0.0001 mM) promastigotes. Conclusions: Despite the effectiveness of exogenous PpIX on promastigotes and the production of PpIX by human infected cells, treatment with ALA or MAL before irradiation was unable to completely destroy L. (L.) infantum or L. (V.) panamensis intracellular amastigotes.

Introducción. El tratamiento fotodinámico con ácido 5-aminolevulínico como inductor de la protoporfirina IX (ALA-PpIX) constituye una alternativa interesante en el tratamiento de la leishmaniasis cutánea. Objetivo. Evaluar la producción de protoporfirina IX (PpIX) a partir de la administración de ALA o MAL y el efecto de la PDT con ALA a nivel celular en células THP-1 no infectadas e infectadas con Leishmania ( Viannia ) panamensis o Leishmania ( Leishmania ) infantum (syn. Leishmania chagasi ). Materiales y métodos. La producción de protoporfirina IX y su ‘colocalización´ mitocondrial se evaluaron mediante microscopía ‘confocal´. Se evaluó la toxicidad celular después del tratamiento con los compuestos y la aplicación de irradiación de luz (597-752 nm) en una fluencia de 2,5 J/cm 2 mediante el empleo de la prueba colorimétrica con metil-tiazol-tetrazolio (MTT) en las células, y de métodos microscópicos estándar en los parásitos. Los resultados se expresaron como la concentración del compuesto activo en el 50 % de las células o parásitos (CC 50 o CI 50 ). Resultados. El ácido aminolevulínico o el metil-5-aminolevulinato indujeron la protoporfirina IX endógena en células humanas, y se observó fluorescencia de color rojo en las mitocondrias. La actividad del ácido aminolevulínico y del metil-5-aminolevulinato utilizados con terapia fotodinámica fue similar en las células THP-1 (CC 50 0,16±0,01 mM y 0,33±0,019 mM, respectivamente) y, aparentemente, no inhibió los parásitos en las células infectadas, en comparación con los controles. El tratamiento exógeno con protoporfirina IX y terapia fotodinámica fue tóxico para las células THP-1 (CC 50 0,00032 ±0,00002 mM) y para los promastigotes de L. (L .) infantum (IC 50 0,003±0,0001 mM) y L. ( V .) panamensis (CI 50 0,024±0,0001 mM). Conclusiones. A pesar de la ‘fotoactividad´ del tratamiento con protoporfirina IX en promastigotes y de su producción después del tratamiento con ácido aminolevulínico y metil-5-aminolevulinato en las células infectadas con Leishmania , no se observó daño en los amastigotes presentes en las células de L. ( L .) infantum o L . ( V .) panamensis .

Manual and Rotary Instrumentation Ability to Reduce Enterococcus faecalis Associated with Photodynamic Therapy in Deciduous Molars

This aim of this study was to assess the ability of manual or rotary instrumentation associated with photodynamic therapy (PDT) to reduce Enterococcus faecalis using three combinations of light/photosensitizers: toluidine blue O/laser, fuchsin/halogen light and fuchsin/LED. Twenty deciduous molars were selected and contaminated with Enterococcus faecalis (McFarland 0.5 scale). Working length determination was performed by visual method. The teeth were randomly divided into two groups: G1 (n=10): manual instrumentation (Kerr-type files) and G2 (n=10): rotary instrumentation (ProTaper system). The bacteria were collected three times using sterile paper cones compatible with the anatomic diameter of the root canal for 30 s before and after instrumentation and after PDT. The samples were diluted in peptone water, seeded on blood agar plates and incubated in an oven at 37 °C for colony-forming units counting. The decrease of E. faecalis counts after instrumentation and after PDT was compared using the Wilcoxon test, t-test and Kruskal Wallis test. A significant reduction of E. faecalis occurred after manual and rotary instrumentation and after PDT using the three combinations of light/photosensitizer (p<0.05). It may be concluded that both rotary and manual instrumentation reduced E. faecalis. Fuchsin with halogen light or LED irradiation and toluidine blue O with laser irradiation can be used to reduce E. faecalis in root canals of primary molars. PDT can be used as an adjuvant to conventional endodontic treatment.

O objetivo do presente estudo foi avaliar a redução de Enterococcus faecalis após instrumentação manual ou rotatória associada à terapia fotodinâmica (PDT) utilizando 3 combinações luz/fotossensibilizante: azul de toluidina O/laser, fucsina/luz halógena e fucsina/LED. Foram selecionados 20 molares decíduos que foram contaminados com Enterococcus faecalis (escala 0,5 de McFarland). A odontometria foi feita através do método visual. Os dentes foram divididos aleatoriamente em dois grupos: G1 (n=10): instrumentação manual (limas tipo Kerr) e G2 (n=10): instrumentação rotatória (sistema ProTaper). Foram realizadas coletas com cone de papel estéril compatível com o diâmetro anatômico do canal durante 30 s antes e após a instrumentação e a PDT. As amostras foram diluídas em água peptonada, semeadas em placas de agar-sangue e incubadas em estufa a 37 °C para contagem das unidades formadoras de colônias. As comparações antes da redução de E. faecalis após a instrumentação e após a realização da PDT foram realizadas pelo teste de Wilcoxon, teste t e Kruskal Wallis. Houve redução significante de E. faecalis após a instrumentação manual ou rotatória e após realização da PDT com as três combinações de luz/fotossensibilizante (p<0,05). Pode-se concluir que a instrumentação rotatória e manual acarretou a redução de E. faecalis. A fucsina irradiada com luz halógena ou led e o azul de toluidina irradiado com laser podem ser utilizados para redução de E. faecalis do sistema de canais radiculares de molares decíduos. A terapia fotodinâmica pode ser utilizada como coadjuvante ao tratamento endodôntico convencional.

Changes in physiological and behavioral parameters of preterm infants undergoing body hygiene: a systematic review / Las variaciones de los parámetros fisiológicos y de comportamiento de los recién nacidos pretérminos sometidos a baño de inmersión: una revisión sistemática / Variações nos parâmetros fisiológicos e comportamentais de recém-nascidos pré-termo submetidos à higienização corporal: revisão sistemática

Objective To verify the effect of bathing on the body temperature of preterm infants (PTI). Method Systematic review conducted in the following bibliographic electronic sources: Biblioteca Virtual em Saúde/Lilacs (BVS), Cumulated Index of Nursing and Allied Health Literature (CINAHL), Cochrane Library, Google Scholar, PubMed, SCOPUS and Web of Science, using a combination of search terms, keywords and free terms. The review question was adjusted to the PICO acronym (Patient/population, Intervention, Control/comparative intervention, Outcome). The selected publications were evaluated according to levels of evidence and grades of recommendation for efficacy/effectiveness studies, as established by the Joanna Briggs Institute. Results Eight hundred and twenty four (824) publications were identified and four studies met the inclusion criteria, of which three analyzed the effect of sponge baths and the effect of immersion baths. Conclusion Sponge baths showed a statistically significant drop in body temperature, while in immersion baths the body temperature remained stable, although they studied late preterm infants. .

Objetivo Determinar el efecto del baño en la temperatura corporal del recién nacido prematuro. Método Revisión sistemática realizada en las fuentes bibliográficas electrónicas BVS, CINAHL, Cochrane Library, Google Scholar, PubMed, Scopus y Web of Science. Las búsquedas fueron realizadas mediante combinación de descriptores, palabras clave y términos libres y se ajustó la cuestión de la revisión a la estrategia PICO. Las publicaciones seleccionadas se evaluaron de acuerdo con los niveles de evidencia y grados de recomendación para los estudios de eficacia/efetividad establecidos por el Instituto Joanna Briggs. Resultados Se identificaron 824 publicaciones y cuatro atendieron a los criterios de inclusión, de los cuales, tres analizaron el efecto del baño de esponja y uno el efecto del baño de inmersión. Conclusión El baño de esponja mostró una disminución estadísticamente significativa en la temperatura corporal, en cuanto que el baño de inmersión, la temperatura corporal se mantuvo estable, aunque el estudio haya sido realizado con recién nacidos prematuros tardíos.

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Objetivo Verificar o efeito do banho na temperatura corporal de recém-nascidos pré-termo (RNPT). Método Revisão sistemática realizada nas fontes eletrônicas bibliográficas BVS/Lilacs, Cumulative Index of Nursing and Allied Health (CINAHL), Cochrane Library, Google Scholar, PubMed, SCOPUS e Web of Science, utilizando a combinação de descritores, palavras-chave e termos livres. A pergunta da revisão foi ajustada ao acrônimo PICO (Paciente/população, Intervenção, Intervenção controle/comparativa, Desfecho analisado). As publicações selecionadas foram avaliadas de acordo com os níveis de evidência e graus de recomendação para estudos de eficácia/efetividade estabelecidos pelo Instituto Joanna Briggs. Resultados Foram identificadas 824 publicações e quatro estudos atenderam aos critérios de inclusão, dos quais três analisaram o efeito do banho de esponja e um o efeito do banho de imersão. Conclusão O banho de esponja mostrou queda da temperatura corporal estatisticamente significante, enquanto no banho de imersão a temperatura corporal permaneceu estável, embora tenham sido estudados RNPT tardios. .

Robertsonian chromosomes and the nuclear architecture of mouse meiotic prophase spermatocytes

BACKGROUND: The nuclear architecture of meiotic prophase spermatocytes is based on higher-order patterns of spatial associations among chromosomal domains from different bivalents. The meiotic nuclear architecture depends on the chromosome characteristics and consequently is prone to modification by chromosomal rearrangements. In this work, we consider Mus domesticus spermatocytes with diploid chromosome number 2n = 40, all telocentric, and investigate a possible modification of the ancestral nuclear architecture due to the emergence of derived Rb chromosomes, which may be present in the homozygous or heterozygous condition. RESULTS: In the 2n = 40 spermatocyte nuclei random associations mediated by pericentromeric heterochromatin among the 19 telocentric bivalents ocurr at the nuclear periphery. The observed frequency of associations among them, made distinguishable by specific probes and FISH, seems to be the same for pairs that may or may not form Rb chromosomes. In the homozygote Rb 2n = 24 spermatocytes, associations also mediated by pericentromeric heterochromatin occur mainly between the three telocentric or the eight metacentric bivalents themselves. In heterozygote Rb 2n = 32 spermatocytes all heterochromatin is localized at the nuclear periphery, yet associations are mainly observed among the three telocentric bivalents and between the asynaptic axes of the trivalents. CONCLUSIONS: The Rb chromosomes pose sharp restrictions for interactions in the 2n = 24 and 2n = 32 spermatocytes, as compared to the ample possibilities for interactions between bivalents in the 2n = 40 spermatocytes. Undoubtedly the emergence of Rb chromosomes changes the ancestral nuclear architecture of 2n = 40 spermatocytes since they establish new types of interactions among chromosomal domains, particularly through centromeric and heterochromatic regions at the nuclear periphery among telocentric and at the nuclear center among Rb metacentric ones.

Interaction of human genes WT1 and CML28 in leukemic cells / 华中科技大学学报（医学）（英德文版）

The molecular pathogenesis of leukemia is poorly understood. Earlier studies have shown both Wilms' tumor 1 suppressor gene (WT1) and CML28 abnormally expressed in malignant diseases of the hematopoietic system and WT1 played an important role in leukemogenesis. However, the relationship between molecular CML28 and WT1 has not been reported. Here we described the use of small interfering RNA (siRNA) against WT1 and CML28 in leukemic cell line K562 to examine the interaction between CML28 and WT1. WT1 and CML28 gene expression in transfected K562 cells was detected by using RQ-PCR and Western blotting. K562 cells transfected with WT1-siRNA could greatly decrease both mRNA and protein expression levels of WT1 and CML28. In contrast, CML28-siRNA did not exert effect on WT1. Further, subcellular co-localization assay showed that the two proteins could co-localize in the cytoplasm of K562 cells, but WT1/CML28 complexes were not detected by using immunoprecipitation. It was suggested that there exists the relationship between CML28 and WT1. CML28 may be a downstream target molecule of WT1 and regulated by WT1, which will provide important clues for further study on the role of CML28 and WT1 in leukemic cells.

Extraction of cytoskeletons and associated proteins using subcellular proteome fractionation technique / 南方医科大学学报

<p><b>OBJECTIVE</b>To develop a method for extracting cytoskeletons and cytoskeleton-associated proteins for proteomic analysis.</p><p><b>METHODS</b>A subcellular sequential proteome extraction method was exploited. The extraction procedure was optimized and controlled according to observed cell morphology changes and one- and two-dimensional electrophoresis images. The extraction efficiency and selectivity were evaluated by Western blotting and mass spectrometry.</p><p><b>RESULTS</b>Four extracted fractions clearly displayed distinct patterns. Western blotting detected the fraction-marker proteins FAK, integrin-β1, histone H1 and cytokeratin 19 only in their expected fractions. About 90% of the protein spots in the cytoskeleton fraction were identified by mass spectrometry as cytoskeleton and/or its associated proteins.</p><p><b>CONCLUSION</b>The subcellular proteome sequential fractionation method facilitates the detection of proteins of low abundance and shows a high reproducibility and selectivity, and thus can serve as an ideal pre-fractionation method prior to two-dimensional electrophoresis.</p>

In vitro reduction of zearalenone to beta-zearalenol by rainbow trout [Oncorhynchus mykiss] hepatic microsomal and post-mitochondrial subfractions

Mycoestrogen zearalenone [ZEA] is found in human foods and animal feeds. Its estrogenic potency mainly depends on its biotransformation fate. The hepatic biotransformation of ZEA in rainbow trout was investigated in this study. Various concentrations of ZEA were separately incubated with the hepatic microsomal and post-mitochondrial sub-fractions in the presence of NADPH, and the metabolites were determined by means of HPLC. Moreover, the rate of glucuronidation for ZEA and its reduced metabolites were estimated in the presence of uridine diphosphate glucuronic acid. beta-zearalenol [beta-ZOL] was found to be the major metabolite of ZEA by both sub-cellular fractions. The enzymatic kinetics analyses indicated that the alpha-ZOL and beta-ZOL production by microsomal fraction were 8- and 2-fold higher than those by post-mitochondrial fraction, respectively. High percentages of ZEA and its metabolites are conjugated with glucuronic acid at the lower concentrations. Data suggest that the hepatic biotransformation of ZEA in rainbow trout resulted in its detoxification as the main metabolite tends to be beta-ZOL with weak estrogenic property. Moreover, at certain concentrations, the produced metabolites are entirely conjugated with glucuronic acid, which may consequently cause a prolonged duration of action due to entero-hepatic cycle

Carboxymethylpachymaran enhances immunologic function of dendritic cells cultured in two kinds of hepatoma carcinoma cell line's supernatant via nuclear factor κ B/Rel pathway / 中国结合医学杂志

<p><b>OBJECTIVE</b>To study the immunologic function of dendritic cells (DCs) cultured in two kinds of hepatoma cell line's supernatant and the enhancing effects of carboxymethylpachymaran (CMP) on DCs.</p><p><b>METHODS</b>DCs were harvested after stimulation by granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin (IL)-4 from umbilical cord blood using density-gradient centrifugation method. Cultured supernatant of two hepatoma cell lines (HepG2 and HepG2.2.15) were collected for condition medium (CM) according to a volume ratio of supernatant to incomplete RPMI-1640 medium, which was 3:1. CMP was dissolved in incomplete RPMI-1640 medium. Experimental groups were divided according to the culture medium, either CM or with CMP in it. DCs subsets CD83, CD86, CD1a, and d-related human leukocyte antigens (HLA-DR) were analyzed by flow cytometry. The proliferation ability of allogeneic T cells in mixed lymphocyte reaction (MLR) stimulated by DCs was examined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) analysis. IL-12p70, interferon-γ (IFN-γ), and nuclear factor κB (NF-κB) were detected by enzyme-linked immunosorbent assay analysis.</p><p><b>RESULTS</b>The proliferation of lymphocytes and secreting level of IL-12 and expression of phenotype of DCs cultured in two kinds of CM were lower than those of normal group (P <0.01). Compared with the normal group, groups treated with CMP showed a higher expression level of DCs subsets, lymphocyte reproductive activity, as well as IL-12 and IFN-γ secretion levels. Groups treated with CMP also demonstrated higher levels of DCs phenotype expression and IL-12 and IFN-γ secretion in supernatant of MLR and higher lymphocyte reproductive activity compared with CM group (P <0.05). Compared with the normal group, the expression level of NF-κB in DCs nuclear was higher in CMP groups but lower in two CM groups (P <0.05). After CMP was added, the NF-κB expression levels of two CM groups were increased compared with levels before CMP was added (P <0.05). However, there was no significant difference between the two CM groups (P >0.05).</p><p><b>CONCLUSIONS</b>Two kinds of hepatoma cell line's supernatant can inhibit the immunologic function of DCs. This suppressive effect may be related to the inhibition of NF-κB/Rel pathway. CMP may up-regulate the DCs function by activating the NF-κB/Rel pathway.</p>

Subcellular localization and resistance to Botrytis cinerea of a new type lipid transfer protein AtDhyPRP1 from Arabidopsis thaliana / 生物工程学报

Genetic transformation was adopted to analyze the subcellular localization and the resistance to fungal pathogens of Arabidopsis lipid transfer protein AtDHyPRP1. The coding sequence of AtDHyPRP1 amplified by PCR from Ws ecotype was used to construct the plant binary expression vector pRI101-AN-AtDHyPRP1 and the fusion expression vector pCAMBIA1302-AtDHyPRP1-GFP. Transgenic tobacco and Arabidopsis plants were produced by leaf disc and floral dip protocols, respectively. AtDHyPRP1 could improve the resistance of tobacco to Botrytis cinerea remarkably and the infection sites on transgenic tobacco leaves accumulated large amounts of H2O2. Observation under laser scanning confocal microscope showed that AtDHyPRP1 was localized to cell surface. It suggested that AtDHyPRP1 might play special function after secretion to outside of the cell and was involved in plant defense system against pathogens.

Effect of chinese herbal drug-containing serum for activating-blood and dispelling-toxin on ox-LDL-induced inflammatory factors' expression in endothelial cells / 中国结合医学杂志

<p><b>OBJECTIVE</b>To investigate the effects of drug-containing serum of Chinese herbal compound, Xiongshao Capsule (, XS, for activating-blood) and Huanglian Capsule (, HL, for dispellingtoxin) on the oxidized low-density lipoprotein (ox-LDL)-induced inflammatory factors in human umbilical vein endothelial cells (HUVECs).</p><p><b>METHODS</b>Thirty-two rats were randomly divided into four groups: the blank control group treated with distilled water, the positive control group treated with simvastatin (1.8 mg/kg), the test group I treated with Chinese herbal compound of XS (0.135 g/kg), and the test group II treated with Chinese herbal compound of XS (0.135 g/kg) and HL (0.135 g/kg). All the treatments were administered for 7 successive days by gastrogavage. Rats' blood serum was harvested 1 h after the last administration to prepare respective drugcontaining serum. HUVECs were exposed to ox-LDL (100 μg/mL) to induce cell injury model and incubated with corresponding drug-containing serum for 24 h. Untreated HUVECs were set for blank control. Levels of interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), and soluble intercellular adhesion molecule-1 (sICAM-1) in supernatant of cultured HUVECs were determined by enzyme-linked immunosorbent assay (ELISA). HUVEC surface expressions of ICAM-1 and E-selectin were determined by flow cytometry.</p><p><b>RESULTS</b>Levels of IL-6, TNF-α, and sICAM-1 in the supernatant of HUVECs as well as the cell surface expressions of ICAM-1 and E-selectin significantly increased after 24-h ox-LDL stimulation (P<0.01), while the abnormal elevations, except sICAM-1 in the test group I, were all reduced in the treated groups (the positive control and the two test groups) significantly (P<0.01 or P<0.05). Besides, the effect in the test group II seemed somewhat higher than that in the test group I but with no statistical significance (P>0.05).</p><p><b>CONCLUSION</b>Drug-containing serum of XS plus HL has a certain inhibitory effect on the vascular endothelial inflammation response induced by ox-LDL.</p>

Salvianolic acid B inhibits the TLR4-NFκB-TNFα pathway and attenuates neonatal rat cardiomyocyte injury induced by lipopolysaccharide / 中国结合医学杂志

<p><b>OBJECTIVE</b>To investigate the role of the TLR4-NFκB-TNFα inflammation pathway on: lipopolysaccharide (LPS)-induced neonatal rat cardiomyocyte injury and the possible protective effects of salvianolic acid B (Sal B).</p><p><b>METHODS</b>Wistar rat (1-2 days old) cardiomyocytes were isolated and cultured. Sal B 10(-5)mol/L, 10(-6)mol/L and 10(-7)mol/L were pre-treated for 6 h in the culture medium. LPS (1 μg/mL) was added to mol/the culture medium and kept for 6 h to induce inflammation injury. The concentration of lactate dehydrogenase (LDH) in the supernatant was detected by spectrophotometry. The concentrations of tumor necrosis factor α (TNFα) and heat shock protein 70 (HSP70) in the supernatant were detected by enzyme linked immunosorbent assay. The protein expressions of toll, such as receptor 4 (TLR4) and nuclear factor kappa B (NFκB) were detected by immunohistochemistry. The mRNA expressions of TLR4 and NFκB were detected by real-realtime reverse transcription polymerase chain reaction (RT-PCR).</p><p><b>RESULTS</b>(1) The concentrations of LDH and: TNFα in the LPS control group were significantly higher than those in the control group (561.41±67.39 U/L and 77.94±15.08 pg/mL, versus 292.13±26.02 U/L and 25.39±16.53 pg/mL, respectively, P<0.01, P<0.05). Compared with the LPS control group, the concentrations of LDH and TNFα were significantly decreased in the Sal B 10(-5)mol/L pre-treated group (451.76±83.96 U/L and 34.00±10.38 pg/mL, respectively, P<0.05). (2) The TLR4 and NFκB protein expression area in the LPS control group were significantly higher than those in the control group (1712.41±410.12 μm(2) and 2378.15±175.29 μm(2), versus 418.62±24.42 μm(2) and 1721.74±202.87 μm(2), respectively, P<0.01). The TLR4 and NFκB protein expression internal optical density (IOD) values in the LPS control group were also significantly higher than those in the control group (3.06±0.33 and 7.20±1.04, versus 0.91±0.21 and 4.24±0.48, respectively, P<0.05 and P<0.01). Compared with the LPS control group, the TLR4 and NFκB protein expression areas were significantly decreased in the Sal B 10(-5)mol/L pre-treated group (1251.54±133.82 μm(2) and 1996.37±256.67 μm(2), respectively, P<0.05), the TLR4 and NFκB protein expression IOD values were also significantly decreased in the Sal B 10(-5)mol/L pre- mol/pretreated group (1.92±0.28 and 5.17±0.77, respectively, treated P<0.05). (3) The TLR4 and NFκB mRNA expressions (2(-ΔΔ)CT value) in the LPS control group were significantly higher than those in the control group (3.16±0.38 and 5.03±0.43 versus 1.04±0.19 and 1.08±0.21, respectively, P<0.01). Compared with the LPS control group, the TLR4 and NFκB mRNA expressions (2(-ΔΔ) -CT value) were significantly decreased in the Sal B 10(-5)mol/L pre- mol/pretreated group (1.34±0.22 and 1.74±0.26, respectively, treated P<0.05). The concentration of HSP70 did not show any <statistical differences in all groups (P>0.05).</p><p><b>CONCLUSIONS</b>The TLR4-NFκB-TNFα pathway was quickly activated: and was independent of HSP70 in the early phase of neonatal cardiomyocyte injury induced by LPS. The protective effects of Sal B may be through inhibiting the TLR4-NFκB-TNFα pathway and are dose-dependent.</p>

Effects of Yinian Jiangya Decoction containing serum on cytokines secretion of vascular endothelium of spontaneously hypertensive rats / 中国结合医学杂志

<p><b>OBJECTIVE</b>To observe the effects of Yinian Jiangya Decoction (YNJYD) on cytokine secretion in spontaneoulsy hypertensive rats (SHRs) vascular endothelium.</p><p><b>METHODS</b>Aortic endothelial cells (ECs) were primarily cultured from SHRs; male SD rats were treated with different doses (high, medium, and low doses) of YNJYD, the blood was collected on the 21st day, and then, the serum was separated. ECs were cocultured with the serum for different time courses, and the culture supernatant concentrations of endothelin (ET)-1, nitric oxide (NO), tissue-type plasminogen activator (t-PA), and plasminogen activator inhibitor (PAI-1) were determined by ABC-ELISA methods.</p><p><b>RESULTS</b>ET-1, NO, t-PA, and PAI-1 levels in endothelial cell culture supernatant were increased in a time-dependent manner; YNJYD could significantly elevate NO and t-PA expressions in ECs, while ET-1 and PAI-1 expressions were dramatically decreased; these effects of YNJYD were in a concentration-dependent manner.</p><p><b>CONCLUSION</b>The therapeutic effect of YNJYD on hypertension is attributed to its effect on regulating vessel dilation and blood coagulation, in which ET-1/NO and PAI-1/t-PA are two pairs of pivotal mediators.</p>

Qushi Huayu Decoction (祛湿化瘀方) inhibits protein and gene expression of cathepsin B in HepG2 cells induced by free fatty acids / 中国结合医学杂志

<p><b>OBJECTIVE</b>To study the experimental efficacy of Qushi Huayu Decoction (祛湿化瘀方,QHD) on, protein and gene expression of cathepsin B (ctsb) in HepG2 cells induced by free fatty acids (FFAs).</p><p><b>METHODS</b>The model of HepG2 steatosis and tumor necrosis factor-α (TNF-α) secretion was induced by long-chain FFAs. HepG2 cells were divided into 4 groups: control group (group C), model group (group M), low-dose QHD group (group L) and high-dose QHD group (group H). Long-chain FFAs were added to groups M, L and H. The 10% blank-control serum was added to group C and M, while 5% and 10% QHD-containing sera were added to group L and H, respectively. The levels of serum TNF-α and cellular triglyceride (TG) were detected. Cellular p-IκB and ctsb expression were detected using Western blot and PCR. The expression and distribution of ctsb were observed by immunofluorescence.</p><p><b>RESULTS</b>After incubating with FFA for 24 h, TG deposition in HepG2, TNF-α content in cell supernatant, the protein expression of cellular ctsb and P-IκB, as well as mRNA expression of ctsb increased markedly in group M compared with group C (P<0.05, P<0.01). Compared with group M, TG deposition, the expression of cellular ctsb, P-IκB and ctsb mRNA in groups L and H, as well as TNF-α content in group H, decreased significantly (P<0.05). Cell immunochemical fluorescence studies showed that ctsb was released from lysosomes and distributed in the cytoplasm extensively and diffusedly after being stimulated with FFA. In this study, these above-mentioned changes were inhibited markedly in groups L and H.</p><p><b>CONCLUSION</b>QHD might have a direct inhibitory effect on the ctsb target in the FFA-ctsb-TNFα pathway of hepatic lipotoxicity.</p>

Apoptosis as pathogenic mechanism of infection with vesicular stomatitis virus: evidence in primary bovine fibroblast cultures

To determine whether fibroblasts from Blanco Orejinegro cattle, exhibit any level of resistance to infection against vesicular stomatitis virus (VSV) serotypes Indiana (VSV-I) or New Jersey (VSV-NJ), 30 fibroblast cultures were phenotyped to evaluate their resistance/susceptibility. Thirty three % of Blanco Orejinegro fibroblast cultures were classified as very resistant, 50% as resistant, and 17% as susceptible to VSV-I infection, whereas 20% were classified as very resistant, 50% as resistant and 30% as susceptible to VSV-NJ infection. Therefore, there appears to be a large variation in phenotypic polymorphism among the fibroblasts to infection by VSV. To elucidate the mechanisms responsible for this diversity, we searched for a possible relationship between resistance/ susceptibility and production of factors wi th antiviral activity; however fibroblasts did not secrete factors with antiviral activity. We examined also whether apoptosis where induced by infection and its correlation with the polymorphism of resistance/susceptibility to VSV. Using morphological analyses, hypoploidy measurements, and level of phosphatidyl serine expression, high levels of apoptosis were measured in VSV infected fibroblasts. However, no correlation exists between apoptosis and the category of resistance/susceptibility to infection, indicating that apoptosis is a pathogenic mechanism of VSV.