Protective effects of total saponins from Panax japonicus against high-fat diet-induced testicular Sertoli cell junction damage in mice / 南方医科大学学报

OBJECTIVE@#To investigate the protective effects of total saponins from Panax japonicus (TSPJ) against high-fat dietinduced testicular Sertoli cell junction damage in mice.@*METHODS@#Forty male C57BL/6J mice were randomized into normal diet group, high-fat diet group, and low-dose (25 mg/kg) and high-dose (75 mg/kg) TSPJ treatment groups (n=10). The mice in the normal diet group were fed a normal diet, while the mice in the other groups were fed a high-fat diet. After TSPJ treatment via intragastric administration for 5 months, the testes and epididymis of the mice were collected for measurement of weight, testicular and epididymal indices and sperm parameters. HE staining was used for histological evaluation of the testicular tissues and measurement of seminiferous tubule diameter and seminiferous epithelium height. The expression levels of ZO-1, occludin, claudin11, N-cadherin, E-cadherin and β-catenin in Sertoli cells were detected with Western blot, and the localization and expression levels of ZO-1 and β-catenin in the testicular tissues were detected with immunofluorescence assay. The protein expressions of LC3B, p-AKT and p-mTOR in testicular Sertoli cells were detected using double immunofluorescence assay.@*RESULTS@#Treatment with TSPJ significantly improved high-fat diet-induced testicular dysfunction by reducing body weight (P < 0.001), increasing testicular and epididymal indices (P < 0.05), and improving sperm concentration and sperm viability (P < 0.05). TSPJ ameliorated testicular pathologies and increased seminiferous epithelium height of the mice with high-fat diet feeding (P < 0.05) without affecting the seminiferous tubule diameter. TSPJ significantly increased the expression levels of ZO-1, occludin, N-cadherin, E-cadherin and β-catenin (P < 0.05) but did not affect claudin11 expression in the testicular tissues. Immunofluorescence assay showed that TSPJ significantly increased ZO-1 and β-catenin expression in the testicular tissues (P < 0.001), downregulated LC3B expression and upregulated p-AKT and p-mTOR expressions in testicular Sertoli cells.@*CONCLUSION@#TSPJ alleviates high-fat diet-induced damages of testicular Sertoli cell junctions and spermatogenesis possibly by activating the AKT/mTOR signaling pathway and inhibiting autophagy of testicular Sertoli cells.

Vitamin D maintains E-cadherin intercellular junctions by downregulating MMP-9 production in human gingival keratinocytes treated by TNF-α

PURPOSE: Despite the well-known anti-inflammatory effects of vitamin D in periodontal health, its mechanism has not been fully elucidated. In the present study, the effect of vitamin D on strengthening E-cadherin junctions (ECJs) was explored in human gingival keratinocytes (HGKs). ECJs are the major type of intercellular junction within the junctional epithelium, where loose intercellular junctions develop and microbial invasion primarily occurs. METHODS: HOK-16B cells, an immortalized normal human gingival cell line, were used for the study. To mimic the inflammatory environment, cells were treated with tumor necrosis factor-alpha (TNF-α). Matrix metalloproteinases (MMPs) in the culture medium were assessed by an MMP antibody microarray and gelatin zymography. The expression of various molecules was investigated using western blotting. The extent of ECJ development was evaluated by comparing the average relative extent of the ECJs around the periphery of each cell after immunocytochemical E-cadherin staining. Vitamin D receptor (VDR) expression was examined via immunohistochemical analysis. RESULTS: TNF-α downregulated the development of the ECJs of the HGKs. Dissociation of the ECJs by TNF-α was accompanied by the upregulation of MMP-9 production and suppressed by a specific MMP-9 inhibitor, Bay 11-7082. Exogenous MMP-9 decreased the development of ECJs. Vitamin D reduced the production of MMP-9 and attenuated the breakdown of ECJs in the HGKs treated with TNF-α. In addition, vitamin D downregulated TNF-α-induced nuclear factor kappa B (NF-κB) signaling in the HGKs. VDR was expressed in the gingival epithelium, including the junctional epithelium. CONCLUSIONS: These results suggest that vitamin D may avert TNF-α-induced downregulation of the development of ECJs in HGKs by decreasing the production of MMP-9, which was upregulated by TNF-α. Vitamin D may reinforce ECJs by downregulating NF-κB signaling, which is upregulated by TNF-α. Strengthening the epithelial barrier may be a way for vitamin D to protect the periodontium from bacterial invasion.

Skin Barrier and Calcium

Epidermal barrier formation and the maintenance of barrier homeostasis are essential to protect us from the external environments and organisms. Moreover, impaired keratinocytes differentiation and dysfunctional skin barrier can be the primary causes or aggravating factors for many inflammatory skin diseases including atopic dermatitis and psoriasis. Therefore, understanding the regulation mechanisms of keratinocytes differentiation and skin barrier homeostasis is important to understand many skin diseases and establish an effective treatment strategy. Calcium ions (Ca²⁺) and their concentration gradient in the epidermis are essential in regulating many skin functions, including keratinocyte differentiation, skin barrier formation, and permeability barrier homeostasis. Recent studies have suggested that the intracellular Ca²⁺ stores such as the endoplasmic reticulum (ER) are the major components that form the epidermal calcium gradient and the ER calcium homeostasis is crucial for regulating keratinocytes differentiation, intercellular junction formation, antimicrobial barrier, and permeability barrier homeostasis. Thus, both Ca²⁺ release from intracellular stores, such as the ER and Ca²⁺ influx mechanisms are important in skin barrier. In addition, growing evidences identified the functional existence and the role of many types of calcium channels which mediate calcium flux in keratinocytes. In this review, the origin of epidermal calcium gradient and their role in the formation and regulation of skin barrier are focused. We also focus on the role of ER calcium homeostasis in skin barrier. Furthermore, the distribution and role of epidermal calcium channels, including transient receptor potential channels, store-operated calcium entry channel Orai1, and voltage-gated calcium channels in skin barrier are discussed.

The role of ubiquitination and deubiquitination in the regulation of cell junctions

Maintenance of cell junctions plays a crucial role in the regulation of cellular functions including cell proliferation, permeability, and cell death. Disruption of cell junctions is implicated in a variety of human disorders, such as inflammatory diseases and cancers. Understanding molecular regulation of cell junctions is important for development of therapeutic strategies for intervention of human diseases. Ubiquitination is an important type of post-translational modification that primarily regulates endogenous protein stability, receptor internalization, enzyme activity, and protein-protein interactions. Ubiquitination is tightly regulated by ubiquitin E3 ligases and can be reversed by deubiquitinating enzymes. Recent studies have been focusing on investigating the effect of protein stability in the regulation of cell-cell junctions. Ubiquitination and degradation of cadherins, claudins, and their interacting proteins are implicated in epithelial and endothelial barrier disruption. Recent studies have revealed that ubiquitination is involved in regulation of Rho GTPases' biological activities. Taken together these studies, ubiquitination plays a critical role in modulating cell junctions and motility. In this review, we will discuss the effects of ubiquitination and deubiquitination on protein stability and expression of key proteins in the cell-cell junctions, including junction proteins, their interacting proteins, and small Rho GTPases. We provide an overview of protein stability in modulation of epithelial and endothelial barrier integrity and introduce potential future search directions to better understand the effects of ubiquitination on human disorders caused by dysfunction of cell junctions.

A N-glicosilação do EGFR afeta a sinalização intracelular e a estabilidade das Junções Tight em células de câncer colorretal / [ EGFR N-glycosylation affects intracellular signaling and stability of Tight Junctions in Colorectal Cancer Cells]

Introdução: O câncer colorretal (CCR) constitui um importante problema de saúde pública no Brasil e no mundo. Durante a progressão de tumores epiteliais, como o CCR, é frequentemente observada uma desestabilização do complexo juncional apical, composto pelas junções tight (JT) e junções aderentes (JA). O aumento dos níveis de expressão das claudinas, principais proteínas transmembranas constituintes das JT, induz a perda da sua função de barreira e aumenta o potencial maligno das células do CCR. Embora cada vez mais estudos estejam focados em compreender melhor os aspectos moleculares que regulam a estabilidade das TJ, o papel desempenhado por N-glicanos neste processo continua sendo pouco explorado. Objetivo: Assim, o objetivo do presente estudo foi investigar o papel dos N-glicanos na regulação da estabilidade das JT em CCR. Materiais e Métodos: Utilizando a linhagem celular HCT-116 (carcinoma colorretal humano), foi avaliado o efeito do tratamento com dois inibidores de biossíntese de N-glicanos, swainsonina (SW) e tunicamicina (Tun), sobre a estabilidade das JT mediante: citometria de fluxo, Western blot, microscopia eletrônica de transmissão, co-imunoprecipitação e imunofluorescência. A expressão de ácido siálico na superfície celular também foi inibida utilizando análogo fluorinado de ácido siálico (Sia-F). Além disso, foram avaliados o grau de N-glicosilação do Receptor do Fator de Crescimento Epidérmico (EGFR) e os níveis de expressão de claudina-3 (cld-3) em tecidos de pacientes com CCR (n = 21) por imunohistoquímica e Western blot (Protocolo-CEP nº: 84/04). Para este estudo foram consultadas bases de dados internacionais (NCBI e TCGA) a fim de identificar os sítios de N-glicosilação (Asn-XSer/Thr) do EGFR e relatos de mutações nestes sítios em CCR. Utilizando um algoritmo classificador de subtipos de CCR foi avaliada também a expressão de glicogenes e de cld-3 em quatro subtipos moleculares de CCR. Ministério da Saúde Instituto Nacional de Câncer Coordenação de Pós-graduação Resultados: Primeiramente, foi avaliado o efeito de SW e Tun na expressão proteica de vários constituintes das TJ. Foi observado que apenas o tratamento com Tun diminuiu os níveis de expressão de cld-3. A Tun também promoveu, concomitantemente, diminuição dos níveis de fosforilação de proteínas na faixa de peso molecular da cld-3 (22kDa) e reorganização da sua localização celular do citoplasma à membrana. Os resultados mostraram que, após o tratamento com Tun, as células HCT-116 desenvolveram contatos celulares mais estabelecidos. Além disso, a deglicosilação do EGFR promoveu redução dos seus níveis de expressão na membrana celular e também diminuiu tanto os seus níveis de ativação quanto a fosforilação das proteínas downstream ERK e AKT. Foi observado que o tratamento com Sia-F diminuiu tanto a ativação de EGFR quanto os níveis de expressão de cld-3. Os resultados mostraram também que tumores de pacientes com CCR, que expressavam maiores níveis proteicos de cld-3 do que os respectivos tecidos normais adjacentes, exibiram também um maior grau de N-glicosilação do EGFR. Finalmente, não foram encontrados relatos de mutações em sítios de N-glicosilação do EGFR em CCR, porém, foram encontradas diferenças na expressão de glicogenes e de cld-3 entre os quatro subtipos moleculares de CCR. Conclusão: Conjuntamente, os resultados demonstram que a N-glicosilação do EGFR desempenha um papel na regulação da cld-3, contribuindo para uma melhor compreensão da biologia do CCR. Estes achados também sugerem um potencial significado clínico de alterações no padrão de N-glicosilação do EGFR.

Substrate roughness induces the development of defective E-cadherin junctions in human gingival keratinocytes

PURPOSE: The entry of bacteria or harmful substances through the epithelial seal of human gingival keratinocytes (HGKs) in the junctional epithelium (JE) is blocked by specialized intercellular junctions such as E-cadherin junctions (ECJs). However, the influence of roughened substrates, which may occur due to apical migration of the JE, root planing, or peri-implantitis, on the development of the ECJs of HGKs remains largely unknown. METHODS: HGKs were cultured on substrates with varying levels of roughness, which were prepared by rubbing hydrophobic polystyrene dishes with silicon carbide papers. The activity of c-Jun N-terminal kinase (JNK) was inhibited with SP600125 or by transfection with JNK short hairpin RNA. The development of intercellular junctions was analyzed using scanning electron microscopy or confocal laser scanning microscopy after immunohistochemical staining of the cells for E-cadherin. The expression level of phospho-JNK was assessed by immunoblotting. RESULTS: HGKs developed tight intercellular junctions devoid of wide intercellular gaps on smooth substrates and on rough substrates with low-nanometer dimensions (average roughness [Ra]=121.3±13.4 nm), although the ECJs of HGKs on rough substrates with low-nanometer dimensions developed later than those of HGKs on smooth substrates. In contrast, HGKs developed short intercellular junctions with wide intercellular gaps on rough substrates with mid- or high-nanometer dimensions (Ra=505.3±115.3 nm, 867.0±168.6 nm). Notably, the stability of the ECJs was low on the rough substrates, as demonstrated by the rapid destruction of the cell junction following calcium depletion. Inhibition of JNK activity promoted ECJ development in HGKs. JNK was closely associated with cortical actin in the regulation of ECJs in HGKs. CONCLUSIONS: These results indicate that on rough substrates with nanometer dimensions, the ECJs of HGKs develop slowly or defectively, and that this effect can be reversed by inhibiting JNK.

Expression Patterns of the Proteins Associated with Cell Junctions in Mouse Testes / 中国医学科学院学报

<p><b>OBJECTIVE</b>To study on the expression patterns of proteins associated with cell junctions in the developing mouse testes.</p><p><b>METHOD</b>The expression levels of reproductive related cell lines spermatogonia cell line GC1 spg, spermatocyte cell line GC2 spg, leydig cell line TM3, and sertoli cell line TM4, primary sertoli cells, and 1-6-week mouse testes were analyzed using Western blot.</p><p><b>RESULTS</b>The sertoli cell junction-associated membrane proteins adhesion molecule A, Occludin and Claudin, and the sertoli-germ cell junction-associated membrane proteins junctional adhesion molecule C, Nectin-3, and E-cadherin were stage-specific in the seminiferous tubules in the mouse testes. The adaptor proteins associated with cell juctions zonula occludens-1, zonula occludens-2, Afadin, Β-catenin, and CD2-associated protein were not stage-specific in the seminiferous tubules in the mouse testes.</p><p><b>CONCLUSIONS</b>In the seminiferous tubules in the mouse testes, the membrane proteins associated with cell junctions are stage-specific. However, the expressions of adaptor proteins associated with cell junctions do not obviously change.</p>

Importância das comunicações intercelulares para o desenvolvimento de folículos ovarianos / Intercellular communications in ovarian follicles

Durante a foliculogênese em mamíferos, ocorre um longo e complexo processo no qual o oócito adquire a competência necessária para a fecundação. Nesse processo ocorre uma comunicação metabólica bidirecional entre os oócitos e as células somáticas dentro do folículo que garante substratos para o oócito em desenvolvimento. Essa comunicação é mediada pelas junções celulares (junções comunicantes e junções aderentes) presentes nas projeções transzonais. As junções celulares e moléculas de adesão são responsáveis principalmente por promover a adesão entre as células foliculares; mas podem atuar em vias de sinalização celular e na regulação da transcrição gênica nas células somáticas e oócitos. Além disso, as junções comunicantes (junções gap) são canais intermembranares que intermediam a comunicação entre essas células através da passagem de pequenas moléculas. Essas junções comunicantes são compostas por proteínas denominadas conexinas; as conexinas 37 e 43 são as predominantes nos folículos ovarianos. Dessa forma, o conhecimento acerca das junções celulares é de extrema importância para o estudo da foliculogênese. A presente revisão teve como objetivo abordar os principais tipos de junções celulares existentes entre as células foliculares, com destaque para as junções gap e as principais proteínas de membranas (conexinas) presentes nos diferentes estágios do desenvolvimento folicular.

During the mammalian folliculogenesis, a long and complex process occurs, which the oocyte acquires the necessary competence for fecundation. In this process there is a metabolic bidirectional communication among the oocyte and somatic cells inside the follicle, which provides substrates for the oocyte developmental competence. This communication is mediated by cellular junctions (occlusions, adherens and gap junctions) localized in the transzonal projections. Cellular junctions and adhesion mollecules are responsable mainly for promoving the adhesion among follicular cells, however they can act in cellular signaling pathways and in regulation of genic transcription in the follicular cells and oocyte. Moreover, the communication junctions (gap junctions) are intermembrane channels that intermediate the communication among these cells through the passage of small molecules. These gap junctions are composed by connexins, of which the connexins 37 and 43 are the most frequently found in the ovarian follicle. Thus, knowledge of these cellular junctions are of great importance for studying the folliculogenesis process. The aim of this review was to report the main types of cellular junctions localized among the follicular cells, especially the gap junctions and the main membrane proteins (connexins) found in different stages of the follicular development.

Physiological understanding of host-microbial pathogen interactions in the gut

The gut epithelial barrier, which is composed of the mucosal layer and the intestinal epithelium, has multiple defense mechanisms and interconnected regulatory mechanisms against enteric microbial pathogens. However, many bacterial pathogens have highly evolved infectious stratagems that manipulate mucin production, epithelial cell-cell junctions, cell death, and cell turnover to promote their replication and pathogenicity in the gut epithelial barrier. In this review, we focus on current knowledge about how bacterial pathogens regulate mucin levels to circumvent the epithelial mucus barrier and target cell-cell junctions to invade deeper tissues and increase their colonization. We also describe how bacterial pathogens manipulate various modes of epithelial cell death to facilitate bacterial dissemination and virulence effects. Finally, we discuss recent investigating how bacterial pathogens regulate epithelial cell turnover and intestinal stem cell populations to modulate intestinal epithelium homeostasis.

Adequação da assistência pré-natal segundo as características maternas no Brasil / Adequacy of prenatal care according to maternal characteristics in Brazil

OBJETIVO: Verificar o grau de adequação da assistência pré-natal no Brasil e sua associação com características sociodemográficas das mulheres. MÉTODOS: Este estudo nacional de base hospitalar foi realizado com 23 894 mulheres em 2011 e 2012. Os dados foram obtidos a partir de entrevistas com a puérpera e dos cartões de pré-natal. Considerou-se assistência pré-natal adequada aquela iniciada até a 12 semana gestacional, com realização de no mínimo seis consultas (número de consultas corrigido para a idade gestacional no momento do parto), registro no cartão de pré-natal de pelo menos um resultado de cada um dos exames preconizados na rotina de pré-natal e recebimento de orientação para maternidade de referência. Realizou-se regressão logística multivariada para verificar a associação entre características maternas e o grau de adequação da assistência pré-natal. RESULTADOS: Início precoce da atenção pré-natal foi observado em 53,9% das gestantes, número adequado de consultas em 73,2%, registro de pelo menos um exame preconizado em 62,9%, orientação para maternidade de referência em 58,7% e assistência pré-natal global adequada em 21,6%. Menor adequação do pré-natal foi observada em mulheres mais jovens, de pele preta, multíparas, sem companheiro, sem trabalho remunerado, com menos anos de estudo, de classes econômicas mais baixas e residentes nas regiões Norte e Nordeste do país. Após ajuste para características maternas, não foram observadas diferenças entre serviços públicos e privados quanto ao grau de adequação do cuidado pré-natal. CONCLUSÕES: A assistência pré-natal no Brasil alcançou cobertura praticamente universal, mas persistem desigualdades regionais e sociais no acesso a um cuidado adequado. Estratégias para facilitar o ingresso precoce no pré-natal são essenciais.

OBJECTIVE: To verify the degree of adequacy of prenatal care in Brazil and to determine whether it is associated with sociodemographic characteristics of women. METHODS: This nationwide hospital-based study was performed with 23 894 women in 2011 and 2012. Data were obtained from interviews with puerperal women and from the prenatal card recording prenatal care appointments. Adequate prenatal care was defined as that started no later than the 12th gestational week, with performance of at least six consultations (with number of consultations adjusted for gestational age at delivery), record in the prenatal card of at least one result for each of the recommended routine prenatal tests, and guidance regarding the maternity hospital for delivery. Multivariate logistic regression was performed to verify the association between maternal characteristics and the adequacy of prenatal care. RESULTS: Early onset of prenatal care was observed in 53.9% of participants, adequate number of consultations in 73.2%, record of at least one of each recommended test in 62.9%, guidance regarding maternity hospital in 58.7%, and overall adequate prenatal care in 21.6%. Less adequate prenatal care was observed in women who were younger, black, multiparous, who did not have a partner, without paid employment, having fewer years of formal schooling, belonging to lower socioeconomic classes, and living in the North and Northeast of Brazil. After adjustment of maternal characteristics, no differences were observed between public or private health care services regarding adequacy of prenatal care. CONCLUSIONS: Even though the coverage of prenatal care is virtually universal in Brazil, regional and social differences in the access and adequacy of care still persist. The implementation of strategies to facilitate early access to prenatal care is essential.

Inhibitory effect of dutasteride on the expressions of epididymal Claudin1 and β-catenin in male rats / 中华男科学杂志

<p><b>OBJECTIVE</b>To explore the molecular mechanism of dutasteride inhibiting fertility by studying its effects on the expressions of the epididymal epithelial junction proteins Claudin1 and β-catenin in rats.</p><p><b>METHODS</b>Sixteen 3-month-old SD male rats were equally divided into an experimental and a negative control group to be treated intragastrically with dutasteride at 40 mg/kg per day and the same dose of solvent, respectively, for 14 consecutive days. Then, the sperm motility and morphology of the rats were detected by computer-assisted sperm analysis, the serum levels of testosterone (T) and dihydrotestosterone (DHT) measured by ELISA, changes in the tight junction of epididymal cells observed under the transmission electron microscope, the protein and gene expressions of Claudin1 and β-catenin determined by RT-PCR and immunohistochemistry, and the conception rate of the mated female rats calculated.</p><p><b>RESULTS</b>Dutasteride significantly suppressed the serum DHT level, sperm motility, and fertility of the rats (P <0.05). Interspaces between epididymal epithelial cell tight junctions were observed, the volume of epididymal fluid obviously increased, and the expressions of Claudin1 and β-catenin gene and protein remarkably downregulated in the experimental rats (P <0.05).</p><p><b>CONCLUSION</b>Dutasteride can significantly inhibit the fertility of male rats by reducing the serum DHT level, suppressing Claudin1 and β-catenin expressions, and damaging epididymal epithelial cell junctions.</p>

A low-protein diet during pregnancy prevents modifications in intercellular communication proteins in rat islets

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Uterine expression of tight junctions in the Canine uterus / 동물의과학연구지

Tight junctions (TJs) form continuous intercellular contacts in intercellular junctions. TJs involve integral proteins such as occludin (OCLN) and claudins (CLDNs) as well as peripheral proteins such as zona occludens-1 (ZO-1) and junctional adhesion molecules (JAMs). TJs control paracellular transportation across cell-to-cell junctions. Although TJs have been studied for several decades, comparison of the transcriptional-translational levels of these molecules in canine organs has not yet been performed. In this study, we examined uterine expression of CLDNs, OCLN, junction adhesion molecule-A, and ZO-1 in canine. Expression levels of canine uterine TJ proteins, including CLDN1, 2, 4, 5, JAM-A, ZO-1, and OCLN, were measured using reverse transcription PCR, real-time PCR, and Western blotting, whereas TJs distribution was determined by immunohistochemistry. The mRNA and protein expression levels of OCLN, CLDN-1, 4, JAM-1, and ZO-1 were identified in the uterus. Immunohistochemistry demonstrated that TJs were localized to the endometrium and/or myometrium of the uterus. Our results show that canine TJ proteins, including CLDNs, OCLN, JAM-A, and ZO-1, were expressed in the canine uterus. Taken together, these proteins may perform unique physiological roles in the uterus. Therefore, these findings may serve as a basis for further studies on TJ proteins and their roles in the physiological or pathological condition of the canine uterus.

O custo de soluções alcalinas em sessões de hemodiálise ambulatorial: uma análise sobre o desperdício a partir do controle dos processos / The cost of alkaline solutions in ambulatory hemodialysis: an analysis about wasteful from the processes control

Introdução: São escassos estudos dos custos dos insumos consumidos em hemodiálise e, dentre estes gastos, os compostos que compõem o dialisato estão entre os valores considerados como representativos nessa terapia. Contudo, não foram encontrados estudos que orientem sobre o comportamento de custos dessas soluções. Objetivo: O objetivo do artigo é avaliar se há desperdício no consumo de soluções alcalinas em hemodiálise ambulatorial e, consequentemente, a possibilidade de redução no custo a partir da simulação de padronização no processo de estabelecimento do fluxo do dialisato nos períodos entre turnos em sessões de hemodiálise ambulatorial. Métodos: Partindo de um estudo observacional analítico, foi realizada uma simulação de 20 cenários, sendo 10 estabelecidos pela padronização dos processos de controle no fluxo do dialisato nos intervalos das sessões. A combinação dos dados foi realizada tomando por base os preços de três fornecedores de soluções alcalinas líquidas ou em pó. Resultados: Observou-se, dentre os cenários com processos padronizados, uma variação entre 7,7% e 33,3% de economia no custo da solução alcalina (em pó ou líquida), pela redução do desperdício. Conclusão: É possível refrear o desperdício no uso de soluções alcalinas, tanto em pó quanto líquidas e, consequentemente, seus custos, a partir da padronização na redução do fluxo de dialisato durante os intervalos verificados entre os turnos na hemodiálise ambulatorial. Todavia, estes resultados estão condicionados ao comprometimento de profissionais de saúde, principalmente no que tange ao exercício da supervisão e controle das atividades ...

Introduction: There are few studies about costs of inputs used in hemodialysis and among these expenditures, the compounds that make up the dialysate are one of the values considered as representative of this therapy. However, there aren’t costs studies that guiding solutions. Objective: The objective of this article is discuss whether there is wasteful of alkaline solutions in ambulatory hemodialysis and hence the possibility of reduction in cost from the standardization process simulation of establishment of dialysate flow in periods between shifts in hemodialysis outpatients. Methods: Starting from an observational analytic, a simulation was performed twenty case scenarios, which ten cases established by standardizing processes control on the dialysate flow in recession. The combination of data was performed using as a basis the prices of three suppliers of alkali liquid or powder. Results: It was observed among the scenarios with standardized processes, ranging between 7.7% and 33.3% savings in the alkaline solution cost (powder or liquid), by reducing waste. Conclusion: It is possible to restrain the wasteful use of alkaline solutions, both powder and liquid. Consequently, its cost from the patterning on reducing the flow of dialysate during the intervals between shifts observed in the outpatient hemodialysis. However, these results are conditional upon the commitment of health professionals, mainly to supervision exercise and control of activities in quality function deployment. .

Xihuang Pill () induces mesenchymal-epithelial transition and inhibits loss of apical-basal polarity in colorectal cancer cell through regulating ZEB1-SCRIB loop / 中国结合医学杂志

<p><b>OBJECTIVE</b>To investigate the antiproliferative and anti-metastasis effect of Xihuang Pill (, XP) on human colorectal cancer cell and to explore the molecular mechanism by which it produces the effects.</p><p><b>METHODS</b>Highly metastatic human colorectal cancer cell line LoVo was treated with low-, medium-, and highdose XP-containing serum (XP-L, XP-M, XP-H) groups for 48 h, cells intervened with no drug rat serum and PD98059 [extracellular signal-regulated kinase (ERK) inhibitor] as negative and positive controls (NC and PC) groups. Cell proliferation assay was made using cell counting kit-8 (CCK8). The 8 μm pore-size transwell chamber and 4', 6-diamidino-2-phenylindole (DAPI) staining were applied to examine the ability of invasion and migration of the cells. The protein expression of ERK1/2, zinc fifi nger E-box-binding homeobox 1 (ZEB1), Scrib and lethal giant larvae homolog 2 (Lgl2) was detected by Western blotting while the relative mRNA quantity of E-cadherin, N-cadherin, Occludin and junctional adhesion molecule-1 (JAM1) was measured by realtime fluorescent quantitative polymerase chain reaction (RT-qPCR).</p><p><b>RESULTS</b>XP induced a dose-dependent suppression on the proliferation of LoVo cells (P <0.05 or P<0.01), with the inhibition rates varied from 27.30% to 31.08%. Transwell assay showed that when preprocessed with PD98059 and XP-containing serum, the number of cells that passed the filter decreased significantly compared with that of NC group (P <0.05 or P<0.01). Moreover, XP inhibited the protein expression of ERK1/2 and ZEB1 (P <0.05); and up-regulated the protein expression of Scrib and Lgl2 (P <0.05). The mRNA levels of E-cadherin, Occludin and JAM1 of the XP intervened groups and PC group markedly ascended (P <0.05) while that of N-cadherin showed a descending tendency (P>0.05).</p><p><b>CONCLUSION</b>XP intervention suppressed the ability of proliferation, invasion and migration of the LoVo cells. Regulating ZEB1-SCRIB Loop so as to recover epithelial phenotype and apical junctional complex might be one of the mechanisms by which XP produces the anti-metastasis effect.</p>

Transplantation of pedicled autologous sinoatrial node tissue for treatment of complete atrioventricular block in dogs / 南方医科大学学报

<p><b>OBJECTIVE</b>To observe the changes of surface ECG and cell couplings between sinoatrial node cells and myocardial cells following transplantion of pedicled autologous sinoatrial node tissue graft into the right ventricle of a canine model of complete atrioventricular block.</p><p><b>METHODS</b>Ten healthy dogs were randomized into transplantation group and control group. Pedicled autologous sinoatrial node tissue grafts were transplanted into the right ventricle in the transplantation group, while the sinoatrial nodes were only excised in the control group after placement of temporary myocardial pacing wires. The changes of surface ECG were observed at 1, 2, 3 and 4 weeks postoperatively. At 4 weeks, complete atrioventricular block was induced in the dogs by radiofrequency ablation of the His bundle. The heart rate of the dogs in both groups were recorded after the injection of isoproternol (ISO) from the femoral vein, and the transplanted tissue graft was observed under optical and transmission electron microscopes.</p><p><b>RESULTS</b>No significant changes occurred in the surface ECG. All the dogs showed ECG waveforms specific of complete heart block after the ablation, and the ventricular heart rates were similar between the two groups (P>0.05). The ventricular heart rate did not undergo obvious changes after ISO injection (P>0.05). The transplanted pedicled autologous sinoatrial node survived in the dogs and the sinoatrial node cells established desmosome junctions with the myocardial cells, but the number of junctions was not sufficient to support heart pacing.</p><p><b>CONCLUSION</b>Desmosome junction can occur between ventricular myocardial cells and sinoatrial node cells at the edge of transplanted pedicled autologous sinoatrial node tissue.</p>

Role of Ultrastructural Alterations of Intercellular Junction and Tight-junction Proteins in Patients With Gastroesophageal Reflux Disease

No abstract available.

Relevance of Ultrastructural Alterations of Intercellular Junction Morphology in Inflamed Human Esophagus

BACKGROUND/AIMS: Detailed characterization of the ultrastructural morphology of intercellular space in gastroesophageal reflux disease has not been fully studied. We aimed to investigate whether subtle alteration in intercellular space structure and tight junction proteins might differ among patients with gastroesophageal reflux disease. METHODS: Esophageal biopsies at 5 cm above the gastroesophageal junction were obtained from 6 asymptomatic controls, 10 patients with reflux symptoms but without erosions, and 18 patients with erosions. The biopsies were morphologically evaluated by transmission electron microscopy, and by using immunohistochemistry for tight junction proteins (claudin-1 and claudin-2 proteins). RESULTS: The expressions of tight junction proteins did not differ between asymptomatic controls and gastroesophageal reflux disease patients. In patients with gastroesophageal reflux disease, altered desmosomal junction morphology was only found in upper stratified squamous epithelium. Dilated intercellular space occurred only in upper stratified squamous epithelium and in patients with erosive esophagitis. CONCLUSIONS: This study suggests that dilated intercellular space may not be uniformly present inside the esophageal mucosa and predominantly it is located in upper squamous epithelium. Presence of desmosomal junction alterations is associated with increased severity of gastroesophageal reflux disease. Besides dilated intercellular space, subtle changes in ultrastructural morphology of intercellular space allow better identification of inflamed esophageal mucosa relevant to acid reflux.

A cell junction pathology of neural stem cells leads to abnormal neurogenesis and hydrocephalus

Most cells of the developing mammalian brain derive from the ventricular (VZ) and the subventricular (SVZ) zones. The VZ is formed by the multipotent radial glia/neural stem cells (NSCs) while the SVZ harbors the rapidly proliferative neural precursor cells (NPCs). Evidence from human and animal models indicates that the common history of hydrocephalus and brain maldevelopment starts early in embryonic life with disruption of the VZ and SVZ. We propose that a "cell junction pathology" involving adherent and gap junctions is a final common outcome of a wide range of gene mutations resulting in proteins abnormally expressed by the VZ cells undergoing disruption. Disruption of the VZ during fetal development implies the loss of NSCs whereas VZ disruption during the perinatal period implies the loss of ependyma. The process of disruption occurs in specific regions of the ventricular system and at specific stages of brain development. This explains why only certain brain structures have an abnormal development, which in turn results in a specific neurological impairment of the newborn. Disruption of the VZ of the Sylvian aqueduct (SA) leads to aqueductal stenosis and hydrocephalus, while disruption of the VZ of telencephalon impairs neurogenesis. We are currently investigating whether grafting of NSCs/neurospheres from normal rats into the CSF of hydrocephalic mutants helps to diminish/repair the outcomes of VZ disruption.

AKAP12 regulates vascular integrity in zebrafish

The integrity of blood vessels controls vascular permeability and extravasation of blood cells, across the endothelium. Thus, the impairment of endothelial integrity leads to hemorrhage, edema, and inflammatory infiltration. However, the molecular mechanism underlying vascular integrity has not been fully understood. Here, we demonstrate an essential role for A-kinase anchoring protein 12 (AKAP12) in the maintenance of endothelial integrity during vascular development. Zebrafish embryos depleted of akap12 (akap12 morphants) exhibited severe hemorrhages. In vivo time-lapse analyses suggested that disorganized interendothelial cell-cell adhesions in akap12 morphants might be the cause of hemorrhage. To clarify the molecular mechanism by which the cell-cell adhesions are impaired, we examined the cell-cell adhesion molecules and their regulators using cultured endothelial cells. The expression of PAK2, an actin cytoskeletal regulator, and AF6, a connector of intercellular adhesion molecules and actin cytoskeleton, was reduced in AKAP12-depleted cells. Depletion of either PAK2 or AF6 phenocopied AKAP12-depleted cells, suggesting the reduction of PAK2 and AF6 results in the loosening of intercellular junctions. Consistent with this, overexpression of PAK2 and AF6 rescued the abnormal hemorrhage in akap12 morphants. We conclude that AKAP12 is essential for integrity of endothelium by maintaining the expression of PAK2 and AF6 during vascular development.