Effect of different obturation techniques on sealer penetration into dentinal tubules after final irrigation with XP-Endo finisher file

The first objective of this research was to evaluate the effectiveness of XP-Endo Finisher on dentinal tubule penetration of irrigation solution using confocal laser scanning microscopy. The main purpose of this research was to compare the effect of cold lateral condensation, continuous wave obturation and core-carrier based techniques on sealer penetration. Sixty mandibular premolars were prepared and allocated into two experimental groups (n=30) as the final irrigation technique and obturation technique experiment. In the final irrigation technique experiment, final irrigation was performed with XP-Endo Finisher, passive ultrasonic irrigation (PUI) and conventional needle irrigation (CNI) (n=10). The roots in the obturation technique experiment were also assigned into 3 groups and obturated with cold lateral condensation, continuous-wave obturation and core-carrier techniques (n=10). The most effective activation method, which emerged as a result of the first part of this study, was used as the final irrigation method in the obturation technique experiment. Then, all roots were sectioned in 1-mm-thick slices at 3mm from the apex for scanning. In terms of depth and percentage of material penetration, CNI exhibited significantly the lowest values and no significant difference was found between others. Also, there was no significant difference among obturation methods. In conclusion, XP-Endo Finisher and PUI are more effective than CNI on irrigant penetration. Sealer penetration into dentinal tubules is independent of obturation techniques.

El objetivo principal de esta investigación fue evaluar la eficacia de XP- Endo Finisher en la penetración de la solución de irrigación en los túbulos dentinarios mediante microscopía de láser confocal. Se prepararon sesenta premolares mandibulares y se distribuyeron en dos grupos experimentales (n=30) según el tipo de método de evaluación utilizado. En el experimento de la técnica de irrigación final, la irrigación final se realizó con XP-Endo Finisher, irrigación ultrasónica pasiva (PUI) e irrigación con aguja convencional (CNI) (n=10). Las raíces en el experimento de la técnica de obturación también se asignaron en 3 grupos y se obturaron con técnicas de condensación lateral fría, obturación de onda continua y portador de núcleo (n=10). El método de activación más eficaz, que surgió como resultado de la primera parte de este estudio, se utilizó como método de irrigación final en el experimento de la técnica de obturación. Luego, todas las raíces se seccionaron en muestras de 1mm de espesor. En términos de profundidad y porcentaje de penetración del material, CNI exhibió significativamente los valores más bajos y no se encontraron diferencias significativas entre los demás. Además, no hubo diferencias significativas entre los métodos de obturación. En conclusión, XP-Endo Finisher y PUI son más efectivos que CNI en la penetración del irrigante. La penetración del sellador en los túbulos dentinarios es independiente de las técnicas de obturación.

Localization of Diacylglycerol Kinase ζ in the Kidney of Adult Rats: Its Dominant Expression in Collecting Tubules and its Selective Lack in Proximal Tubules / Localización de Diacilglicerol Quinasa ζ en el Riñón de Ratas Adultas: su Expresión Dominante en Túbulos Colectores y su Carencia Selectiva en Túbulos Proximales

SUMMARY: Diacylglycerol kinase (DGK) exerts balancing the intracellular level between two-second messengers, diacylglycerol and phosphatidic acid, by its phosphorylation activity. DGK ζ is often localized in cell nuclei, suggesting its involvement in the regulation of intranuclear activities, including mitosis and apoptosis. The present immunohistochemical study of rat kidneys first revealed no detection levels of DGK ζ -immunoreactivity in nuclei of most proximal tubule epithelia in contrast to its distinct occurrence in cell nuclei of collecting and distal tubules with the former more dominant. This finding suggests that DGK ζ is a key factor regulating vulnerability to acute kidney injury in various renal tubules: its low expression represents the high vulnerability of proximal tubule cells, and its distinct expression does the resistance of collecting and distal tubule cells. In addition, this isozyme was more or less localized in nuclei of cells forming glomeruli as well as in endothelial nuclei of peritubular capillaries and other intrarenal blood vessels, and epithelial nuclei of glomerular capsules (Bowman's capsules) and renal calyces, including intrarenal interstitial cells.

La diacilglicerol quinasa (DGK) ejerce el equilibrio del nivel intracelular entre dos segundos mensajeros, diacilglicerol y ácido fosfatídico, por su actividad de fosforilación. La DGK ζ a menudo se localiza en los núcleos celulares, lo que sugiere su participación en la regulación de las actividades intranucleares, incluidas la mitosis y la apoptosis. El presente estudio inmunohistoquímico en riñones de rata no reveló niveles de detección de inmunorreactividad de DGK ζ en los núcleos de la mayoría de los epitelios de los túbulos proximales, en contraste a la detección en los núcleos celulares de los túbulos colectores y distales, siendo el primero más dominante. Este hallazgo sugiere que DGK ζ es un factor clave que regula la vulnerabilidad a la lesión renal aguda en varios túbulos renales: su baja expresión representa la alta vulnerabilidad de las células del túbulo proximal, y su expresión distinta hace a la resistencia de las células del túbulo colector y distal. Además, esta isoenzima estaba más o menos localizada en los núcleos de las células que forman los glomérulos, así como en los núcleos endoteliales de los capilares peritubulares y otros vasos sanguíneos intrarrenales, y en los núcleos epiteliales de las cápsulas glomerulares (cápsulas de Bowman) y los cálices renales, incluidas las células intersticiales intrarrenales.

Clinicopathological features analysis of focal segmental glomerulosclerosis after kidney transplantation / 器官移植

Objective To investigate the clinicopathological features of recurrent and de novo focal segmental glomerulosclerosis (FSGS) after kidney transplantation. Methods Thirty-four recipients pathologically diagnosed with FSGS by renal allograft biopsy were enrolled in this clinical trial. According to the detection of primary diseases of renal allografts and circulating permeability factors, 34 recipients were divided into the recurrent FSGS group (n=12) and de novo FSGS group (n=22). The differences of clinical indexes and the degree of pathological injury of renal allografts were compared between two groups. Results There was no significant difference in the mesangial hyperplasia score, glomerulosclerosis rate, renal tubular atrophy score, interstitial fibrosis score and podocyte proliferation rate between two groups (all P > 0.05). In the recurrent FSGS group, segmental glomerulosclerosis rate of the recipients was 0.10 (0.08, 0.27), lower than 0.19 (0.13, 0.33) in the de novo FSGS group (P < 0.05). No significant difference was found in the incidence of antibody-mediated rejection, drug-induced renal tubular injury and BK virus infection between two groups (all P > 0.05). The incidence of T cell-mediated rejection in the recurrent FSGS group was 17%, lower than 55% in the de novo FSGS group (P < 0.05). Immunohistochemical staining showed that the infiltrating inflammatory cells in the renal allografts were mainly T lymphocytes. The positive rates of C4d deposition in peripheral capillaries between the recurrent and de novo FSGS groups were 33% (4/12) and 32% (7/22), with no significant difference (P > 0.05). Immunofluorescence results revealed IgM deposition in the segmental glomerulosclerosis area of renal allografts in most cases. Electron microscopy showed extensive fusion or segmental distribution of podocytes in the glomerulus of renal allografts. Conclusions The degree of renal functional injury and the incidence of T cell-mediated rejection in the recurrent FSGS group are lower than those in the de novo FSGS group. Comprehensive analysis of preoperative and postoperative clinical manifestations, laboratory testing and pathological examination of kidney transplant recipients contribute to early diagnosis and treatment of recurrent and de novo FSGS.

Three-dimensional visualization on the relationship between tubules after macula densa and afferent arterioles in mouse kidney / 解剖学报

Objective To establish the spatial course of distal tubule and afferent arterioles after macula densa, and to locate and detect the proteins in the adjacent parts by using three-dimensional visualization technology of microstructure. Methods C57 BL/6J mice were fixed by perfusion and embedded in epon 812. Tissue blocks were cut perpendicular to the longitudinal axis of the kidney. And a total of 720, 2. 5 μm-thick consecutive sections were obtained from the renal capsule to the outer stripe of the renal outer medulla. After aligning the digital microscopic images through computer registration procedures, the tubules and vessels were traced by 3D reconstruction program edited by C Language. Selecting the tissue sections of the contact site and applying the improved immunoperoxidase staining method to detect H

The effectiveness of insulin therapy in the functional and morphological recovery of leydig cells in rats submitted to Diabetes mellitus

The effects of systemic insulin administration at different concentrations on the testicular tissue of diabetic adult rats, induced by streptozotocin, are evaluated by the morphological analysis of spermatogenic process. Twenty-four adult male rats were divided into 1) Control Group: they received citrate buffer, by intraperitoneal injection; 2) Diabetic Group: induced by intraperitoneal injection of streptozotocin (60 mg. kg-1 of body weight); 3) Insulin 50%: induced diabetes treated with half of standard dosage of insulin; 4) Insulin 100%: induced diabetes treated with standard dose of insulin. After eight weeks, animals were weighted and anesthetized; testicles were removed and processed in resin. Body and testicular weight of diabetic rats decreased when compared to that of control. Parameters increased with insulin therapy. Testosterone levels were low in diabetic animals but rates recovered after insulin therapy. Nuclear diameter and volume of Leydig cells decreased in diabetic rats although they significantly increased after insulin therapy. Results showed that the administration of insulin in diabetic rats promoted a protective effect of testicular parenchyma, enhancing efficient recovery on testosterone levels and increase in daily sperm production.

Control of sodium and potassium homeostasis by renal distal convoluted tubules

Distal convoluted tubules (DCT), which contain the Na-Cl cotransporter (NCC) inhibited by thiazide diuretics, undergo complex modulation to preserve Na+ and K+ homeostasis. The lysine kinases 1 and 4 (WNK1 and WNK4), identified as hyperactive in the hereditary disease pseudohypoaldosteronism type 2, are responsible for activation of NCC and consequent hypokalemia and hypertension. WNK4, highly expressed in DCT, activates the SPAK/OSR1 kinases, which phosphorylate NCC and other regulatory proteins and transporters in the distal nephron. WNK4 works as a chloride sensor through a Cl- binding site, which acts as an on/off switch at this kinase in response to changes of basolateral membrane electrical potential, the driving force of cellular Cl- efflux. High intracellular Cl- in hyperkalemia decreases NCC phosphorylation and low intracellular Cl- in hypokalemia increases NCC phosphorylation and activity, which makes plasma K+ concentration a central modulator of NCC and of K+ secretion. The WNK4 phosphorylation by cSrc or SGK1, activated by angiotensin II or aldosterone, respectively, is another relevant mechanism of NCC, ENaC, and ROMK modulation in states such as volume reduction, hyperkalemia, and hypokalemia. Loss of NCC function induces upregulation of electroneutral NaCl reabsorption by type B intercalated cells through the combined activity of pendrin and NDCBE, as demonstrated in double knockout mice (KO) animal models, Ncc/pendrin or Ncc/NDCBE. The analysis of ks-Nedd-4-2 KO animal models introduced the modulation of NEDD4-2 by intracellular Mg2+ activity as an important regulator of NCC, explaining the thiazide-induced persistent hypokalemia.

Evaluation of the dentinal tubule penetration of an endodontic bioceramic sealer after three final irrigation protocols

Introduction: This study aimed to evaluate the dentinal tubule penetration of an endodontic bioceramic sealer, Sealer Plus BC, after three final irrigation protocols. Methods: Thirty distobuccal roots of maxillary molars were selected. Root canal preparation was performed up to an #40.06 instrument (X1 Blue) under 2.5% sodium hypochlorite irrigation. Specimens were randomly divided into three groups (n=10), according to the final irrigation protocol: G-NaOCl (2.5% sodium hypochlorite + PUI), G-SS (0.9% saline solution + PUI) and G-H20 (Deionized water + PUI). After final irrigation protocols, all specimens were irrigated with phosphate buffer solution. Root canal obturation was performed using the single cone technique and Sealer Plus BC, stained with a specific fluorophore. Specimens were transversely sectioned and each root third was evaluated in a confocal scanning laser microscopy. Images obtained were analyzed for sealer penetration in the dentinal tubules. Results: Dentinal tubule penetration of Sealer Plus BC was not observed in any root third, regardless of the final irrigation protocol investigated. Conclusions: Sealer Plus BC dentinal tubule penetration was not observed after none of the protocols tested. Dentinal tubule penetrability of Sealer Plus BC may be related to other factors rather than the final irrigation protocol.

Capacidad de Penetración y Difusión de la Medicación, Intraconducto en Túbulos Dentinales, Conductos Laterales e Istmos. Una Revisión Sistemática

RESUMEN: El objetivo de la terapia endodóntica consiste en tratar o prevenir la periodontitis apical. La efectiva eliminación o disminución de microbiota es esencial para la reparación periapical. Debido a esto, se utiliza el empleo de medicamentos intraconductos para tratar químicamente el sistema de conductos radiculares, y cobra relevancia la capacidad de penetración y difusión de dicho medicamento para entrar en contacto directo con los microorganismos que penetren dentro de los túbulos dentinales, conductos laterales e itsmos. Se realizó una revisión de la literatura en bases de datos electrónicas con los buscadores "Pubmed, Scielo y Google Académico", utilizando palabras clave para identificar la literatura publicada relevante en inglés. El análisis y los criterios de elegibilidad se documentaron de acuerdo con los elementos de informe para revisiones sistemáticas y meta-análisis (Preferred Reporting Items for Systematic Reviews and Meta-Analysis, PRISMA). No se impusieron restricciones sobre la fecha de publicación. Se recogieron datos sobre capacidad de difusión de medicación intraconductos en túbulos dentinales, conductos laterales e itsmos radiculares y se sometieron a análisis descriptivo de datos. La búsqueda bibliográfica en bases de datos electrónicas según los criterios de inclusión arrojó 45 títulos y resúmenes. Entre ellos, 7 artículos fueron seleccionados. Se determinó un rango de penetración del medicamento muy variable, entre 27,7 - 2350 μm y rango de difusión de 200-500 μm. Hay pocos estudios publicados que reporten la capacidad de penetración y difusión de la medicación intraconductos. El vehículo empleado y diámetro de las partículas en la preparación del medicamento y la eliminación de la capa de desecho aumentaría la capacidad de penetración/difusión del medicamento intraconductos dentro del túbulo dentinal, conductos laterales e itsmo.

ABSTRACT: The objective of endodontic therapy is to treat or prevent apical periodontitis. The effective elimination or reduction of the microbiota is essential for periapical repair. Due to this, the use of intracanal drugs is used to chemically treat the root canal system, and the penetration and diffusion capacity of said drug becomes relevant to come into direct contact with the microorganisms that penetrate inside the dentinal tubules, lateral canals and isthmus. A literature review was carried out in electronic databases with the search engines "Pubmed, Scielo and Google Academic", using keywords to identify the relevant published literature in English. The analysis and eligibility criteria were documented according to the reporting items for systematic reviews and meta-analyzes (Preferred Reporting Items for Systematic Reviews and Meta-Analysis, PRISMA). No restrictions were placed on the date of publication. Data on intracanal drug diffusion capacity in dentinal tubules, lateral canals and root isthmus were collected and subjected to descriptive data analysis. The bibliographic search in electronic databases according to the inclusion criteria yielded 45 titles and abstracts. Among them, 7 articles were selected. A highly variable drug penetration range was determined, between 27.7 - 2350 μm and a diffusion range of 200-500 μm. There are few published studies that report the penetration and diffusion capacity of intracanal medication. The vehicle used and the diameter of the particles in the preparation of the drug and the elimination of the waste layer would increase the penetration / diffusion capacity of the drug within the dentinal tubule, lateral canals and isthmus.

Three-dimensional visualization of the renal proximal tubules in developing mice / 解剖学报

Objective Adult proximal tubule ( PT ) is not only the segment most frequently involved in acute renal tubule injury, but also the easiest to repair. It may be consistent with the rapid growth and differentiation mechanism of this segment during the development of the kidney, while the developing information is insufficient. Therefore, we three- dimensional visualized the developing PT to analysis its spatiotemporal morphogenesis. Methods The kidneys were obtained from mice at various developing time point, embryonic day ( E ), postnatal day ( P ). The volume density of Claudin-2 positive PT in the cortex was measured using a stereological method in paraffin sections. After image recording and alignment of the serial sections, the spatial courses of the developing PT were traced and visualized in three dimensions using computer-assisted program. The length of the developing PT was calculated at the same time. Results The volume density of PT in the cortex of PI mice was significantly higher than that in the embryonic stage. Then it experienced a decline ( P3, P5 ), an increase ( start at P7 ) to a stable adult level ( P28 ). The tubular tracing showed that the lengths of developing PT and the number of convolutions of their convoluted part increased with the maturation, but lower than that of adultin E14. 5, E17. 5 and P5 PT in E14. 5 and E17. 5 mice were similar to that of adult with respect to general spatial courses. They were, however, significantly different from adult in the initial direction of PT and the arrangement of the straight part of PT in the medullary rays. While, it was in P5 that the spatial pattern of some PT was gradually approaching to the adult model. Conclusion This study demonstrated that the development of PT was consistent with the kidney development in terms of its volume density in cortex, length and spatial course. It started at the S-shaped body, kept throughout the embryonic period and continued to postnatal, ended at kidney maturation ( P28 ).

Morphological aspects of epididymal microscopy and rete testis in greater Rhea Americana / Aspectos morfológicos da microscopia do epitélio do epidídimo da Rhea americana

ABSTRACT: The purpose of this research was to study the histology and describe the microscopy findings of the epididymis epithelium of greater Rhea americana at three time periods: November 2005 (n=14), December 2006 (n= 20), and May 2007 (n= 20), to observe and compare the differences that occurred. We studied the epididymis from 54 rheas, bred in Santa Maria, Rio Grande do Sul, Brazil. The epididymis were collected during commercial slaughter and fixed in bouin. Optical microscopy was used to measure the cellular structure, types of cells, tubules, and stereological values like the epididymis epithelium diameters, lumen, thickness, and relative volume of the tissue structure. Additionally, electron microscopy was studied. In December 2006 and May 2007, the means of the epididymis tubular diameter were: 79.1 and 58.1 µm, epithelium thickness: 24.0 and 52.2 µm, and lumen diameter: 55.0 and 5.8 µm, respectively. Regarding the volumetric proportion, we reported the following values: epithelium volume 36.2 and 80.4%, lumen without spermatozoon 19.6 and 3.0%, lumen with spermatozoon 5.4 and 0.0%, interstitium 35.4 and 12.0%, blood vessels 3.5 and 4.6%, structures in cellular superficies 1.4 and 0%, lamina 1.4 and 3.2%, and artifacts 0.3 and 1.3%, respectively. The epididymis ducts had a circular form in transverse sections with spermatozoon only in November 2005 and December 2006. The Rhea's epididymis morphology was found to be similar to ostriches, roosters, and Japanese quail. Here, we present data from stereological microscopy (tubular diameter, epithelium thickness, and lumen diameter), volumetric proportion (epithelium, lumen without spermatozoon, lumen with spermatozoon, interstitium, blood vessels, structures in cellular superficies; cilium, estereocilium, and lamina) in this species during the repose and sexual activity period (reproductive season).

RESUMO: O objetivo desta pesquisa foi estudar a histologia e descrever os achados microscópicos do epitélio epidídimo de ema em três períodos: novembro/2005 (n = 14), dezembro/2006 (n = 20) e maio/2007 (n = 20), para observação e comparação das diferenças que ocorreram nesses tempos distintos. Estudamos o epidídimo de 54 animais, criados em Santa Maria, RS. Durante o abate comercial, os epidídimos foram coletados e fixados em Bouin. A microscopia óptica mede a estrutura celular, tipos de células, túbulos e valores estereológicos, como os diâmetros do epitélio do epidídimo, lúmen, espessura, volume relativo da estrutura do tecido. Em dezembro/2006 e maio/2007, as médias do diâmetro tubular do epidídimo foram: 79,1 e 58,1µm, espessura do epitélio: 24,0 e 52,2µm, diâmetro do lúmen: 55,0 e 5,8µm, respectivamente. Em relação à proporção volumétrica, encontramos os seguintes valores: volume epitelial 36,2 e 80,4%, lúmen sem espermatozoide 19,6 e 3,0%, lúmen com espermatozoide 5,4 e 0,0%, intestínio 35,4 e 12,0%, vasos sanguíneos 3,5 e 4,6%, estruturas celulares superfícies 1,4 e 0%, lâmina 1,4 e 3,2%, artefatos 0,3 e 1,3%, respectivamente. Os ductos do epidídimo apresentaram forma circular em cortes transversais com espermatozoide apenas em novembro/2005 e dezembro/2006. A morfologia do epidídimo de Rhea foi semelhante ao avestruz, galo e codornas japonesas. Apresentamos dados de microscopia estereológica (diâmetro tubular, espessura do epitélio, diâmetro do lúmen), proporção volumétrica, em porcentagem (epitélio, lúmen sem espermatozóide, lúmen com espermatozoide, intestório, vasos sanguíneos, estruturas em superfícies celulares - cílio - estereocílio, lâmina e artefatos) nesta espécie nos períodos de repouso e atividade sexual (estação reprodutiva).

Acetaminophen induces alterations to the renal tubular ultrastructure in a rat model of acute nephrotoxicity protected by resveratrol and quercetin / El acetaminofeno induce alteraciones en la ultraestructura tubular renal en un modelo de rata con nefrotoxicidad aguda protegida por resveratrol y quercetina

Acetaminophen (also called paracetamol, or APAP) induced nephrotoxicity is reported after accidental or intentional ingestion of an overdose of the drug. Renal tubular ultrastructural alterations induced by APAP overdose associated with the induction of biomarkers of kidney injury have not been investigated before. Also, we investigated whether the combined polyphenolic anti-inflammatory and antioxidants agents, resveratrol (RES) and quercetin (QUR) can protect against APAP-induced acute kidney injury. The model group of rats received a single dose of APAP (2 g/kg), whereas the protective group of rats was pre-treated for 7 days with combined doses of RES (30 mg/kg) and QUR (50 mg/kg) before being given a single dose of APAP. All rats were then sacrificed one day post APAP ingestion. Harvested kidney tissues were prepared for transmission electron microscopy (TEM) staining and blood samples were assayed for urea, creatinine, and biomarkers of inflammation and oxidative stress. TEM images and blood chemistry analysis showed that APAP overdose induced kidney damage as demonstrated by substantial alterations to the proximal convoluted tubule ultrastructure, and a significant (p<0.05) increase in urea, creatinine, tumor necrosis factor-alpha (TNF-a), and malondialdehyde (MDA) blood levels, which were protected by RES+QUR. These findings indicate that APAP induces alterations to the renal tubular ultrastructure, which is inhibited by resveratrol plus quercetin, which also decreases blood levels of kidney injury biomarkers.

El objetivo de este trabajo fue estudiar la nefrotoxicidad inducida por acetaminofeno (también llamado paracetamol o APAP) después de la ingestión accidental o intencional de una sobredosis de la droga. Las alteraciones ultraestructurales tubulares renales inducidas por sobredosis de APAP asociadas con la inducción de biomarcadores de daño renal no se han investigado. Además, estudiamos si los agentes combinados antiinflamatorios y antioxidantes polifenólicos, el resveratrol (RES) y la quercetina (QUR) pueden proteger contra la lesión renal aguda inducida por APAP. El grupo modelo de ratas recibió una dosis única de APAP (2 g / kg), mientras que el grupo protector de ratas se trató previamente durante 7 días con dosis combinadas de RES (30 mg / kg) y QUR (50 mg / kg) antes de recibir una dosis única de APAP. Todas las ratas se sacrificaron un día después de la ingestión de APAP. Los tejidos renales fueron preparados para el análisis a través de la microscopía electrónica de transmisión (MET). En las muestras de sangre se determinaron la urea, creatinina y los biomarcadores de inflamación y estrés oxidativo. Las imágenes MET y el análisis químico de la sangre mostraron que la sobredosis de APAP inducía daño renal, como lo demuestran las alteraciones sustanciales en la ultraestructura del túbulo contorneado proximal, y además, de un aumento significativo (p <0,05) de la urea, creatinina, factor de necrosis tumoral alfa y niveles sanguíneos de malondialdehído, protegidos por RES + QUR. Estos hallazgos indican que APAP induce alteraciones en la ultraestructura tubular renal, inhibida por el resveratrol más quercetina, que también disminuye los niveles sanguíneos de biomarcadores de daño renal.

The role of renal tubular Injury in the transition from acute kidney injury to chronic kidney disease / 医学研究生学报

Acute kidney injury (AKI) is an acute and critical kidney disease that seriously threatens human health and even life. It can lead to the chronic kidney disease (CKD) and end-stage kidney disease (ESRD), causing global public health problems. At present, the transition mechanism from AKI to CKD has not been fully elucidated, and renal tubular injury may play an important role in it. This paper reviews the role of renal tubular injury in the transition from AKI to CKD including cell dedifferentiation, inflammatory cell infiltration, hypoxia and vascular density decline, mitochondrial dysfunction and oxidative stress, G2/M cell cycle arrest, autophagy, TGFβ signaling pathway and Wnt signaling pathway.

Effect of Fushengong Prescription on Wnt/β-catenin Signaling Pathway in Kidney of Rats with Chronic Renal Failure / 中国实验方剂学杂志

Objective:To observe the effects of Fushengong prescription on secretory glycoprotein （Wnt）/β-serial protein （β-catenin） signaling pathway in kidney of rats with chronic renal failure （CRF），and to further explore its mechanism of releasing the aggregation of extracellular matrix（ECM），inhibiting renal tubule interstitial fibrosis （TIF） and prolonging the progression of CRF. Method:A total of 55 SD male rats were randomly divided into the normal group，the model group，and the low， medium and high dose groups of Fushengong prescription，with 11 rats in each group.The normal group was routinely reared and the other 4 groups of rats were used to establish CRF model with 0.5% adenine fodder， fed them continuously for 21 d. After successful modeling，all model rats were switched to conventional feed. Normal saline （NS） was given the normal group and the model group by 20 mL·kg-1·d-1， the low， middle and high dose groups rats of Fushengong prescription were given intragastric administration Fushengong prescription according to the body weight of 4， 8， 16 g·kg-1，once a day，continuous gavage for 30 d. After the experiment，the pathomorphism change of renal tissues of rats was measured by Masson staining， the expression of Wnt4 and β-catenin mRNA in the kidney tissues were observed by the method of Real-time fluorescence quantitative polymerase chain reaction（Real-time PCR）， the expression of Wnt4，β-catenin and matrix metalloproteinase-7（MMP-7） protein of renal tissues were detected by the methods of Western blot. The expression of Wnt4， β-catenin protein of renal tissues were detected by the methods of immunohistochemistry （IHC）. Result:Compared with normal group，renal tubule interstitial fibrosis of renal tissues increased distinctly and the expression of Wnt4，β-catenin and MMP-7 protein increased significantly in the model group. Wnt4 and β-catenin mRNA also increased significantly in model group（P<0.01）. Compared with model group， the expression of Wnt4， β-catenin and MMP-7 protein in the Fushengong prescription groups decreased obviously （P<0.05）. The expression of Wnt4 and β-catenin mRNA in Fushengong prescription groups also decreased obviously. Conclusion:The mechanism of Fushengong prescription can release the aggregation of ECM，inhibit TIF and delay the progression of CRF，which may be related with the activation of Wnt/β-catenin signal pathway.

Palbociclib induces cell cycle arrest and senescence of human renal tubular epithelial cells / 南方医科大学学报

OBJECTIVE@#To investigate the effect of palbociclib on cell cycle progression and proliferation of human renal tubular epithelial cells.@*METHODS@#Human renal tubular epithelial cell line HK-2 was treated with 1, 5, 10, and 20 μmol/L of palbociclib, and the changes in cell proliferation and viability were examined by cell counting and CCK8 assay. EDU staining was used to assess the proliferation of HK-2 cells following palbiciclib treatment at different concentrations for 5 days. The effect of palbociclib on cell cycle distribution of HK-2 cells was evaluated using flow cytometry. SA-β-Gal staining and C12FDG senescence staining were used to detect senescence phenotypes of HK-2 cells after palbociclib treatment at different concentrations for 5 days. The relative mRNA expression levels of P16, P21, and P53 and the genes associated with senescence-related secretion phenotypes were detected by RT-PCR, and the protein expressions of P16, P21 and P53 were detected by Western blotting.@*RESULTS@#Palbociclib inhibited HK-2 cell proliferation and induced cell cycle arrest in G1 phase. Compared with the control cells, HK-2 cells treated with high-dose (10 μmol/L) palbociclib exhibited significantly suppressed cell proliferation activity, and the inhibitory effect was the most obvious on day 5 (@*CONCLUSIONS@#Palbociclib induces HK-2 cell senescence by causing cell growth arrest and delaying cell cycle progression.

Influence of root canal filling techniques on sealer penetration and gutta percha/sealer ratio: a confocal laser scanning microscopy analysis / Influência das técnicas de obturação do canal radicular na penetração do cimento e na relação guta-percha / cimento: uma análise por microscopia de varredura confocal a laser

Objective: The influence of four root canal filling techniques on the penetration of an endodontic sealer into dentinal tubules and the gutta percha/ sealer ratio (GP/SR) in root canals was evaluated using confocal laser scanning microscopy (CLSM). Material and Methods: Roots of the maxillary central incisors (n=40) were prepared with ProTaper Universal files up to file F5 and assigned to five groups: continuous wave condensation, lateral condensation, single cone, Thermafill®, and negative control group. After root canal filling with gutta-percha and AH26, along with the addition of 0.01% fluorescein, the roots were cut into 2-mm slices. Using CLSM, the specimens were transversely sectioned at 3, 6, and 10 mm from the apex. Results: Sealer penetration was deeper and more frequent at 10 mm than at the 6mm and 3mm for all obturation technique. Penetration was not significantly affected by obturation techniques except single master cone tecnique. Single cone technique demonstrated the lowest sealer penetration at all levels. However, sealer thickness was strongly dependent on obturation technique. Termafill® demostrated superior GP ratio followed by continuous wave condensation, lateral condensation and single cone. Conclusion: In conclusion, the single cone technique resulted in lower sealer penetration than the other techniques, which did not differ significantly from each other. However, sealer thickness was strongly dependent on obturation technique. Termafill® demostrated superior GP ratio followed by continuous wave condensation, lateral condensation and single cone. (AU)

Objetivo: A influência de quatro técnicas de obturação do canal radicular na penetração de um cimento endodôntico nos túbulos dentinários e a relação gutapercha / cimento (GP / CIM) em canais radiculares foram avaliadas por microscopia de varredura confocal a laser (MVCL). Material e Métodos: As raízes de incisivos centrais superiores (n = 40) foram preparadas com limas ProTaper Universal até a lima F5 e distribuídas em cinco grupos: condensação de onda contínua, condensação lateral, cone único, Thermafill® e grupo de controle negativo. Após o preenchimento do canal radicular com guta-percha e AH26, juntamente com a adição de 0,01% de fluoresceína, as raízes foram cortadas em fatias de 2 mm. Usando MVCL, as amostras foram seccionadas transversalmente a 3, 6 e 10 mm do ápice. Resultados: A penetração do cimento foi mais profunda e mais frequente em 10 mm do que nos 6 mm e 3 mm para todas as técnicas de obturação. A penetração não foi significativamente afetada pelas técnicas de obturação, exceto pela técnica de cone mestre único. A técnica de cone único demonstrou a menor penetração do cimento em todos os níveis. No entanto, a espessura do cimento foi fortemente dependente da técnica de obturação. O Termafill® demonstrou uma relação superior de GP, seguida por condensação de onda contínua, condensação lateral e cone único. Conclusão: Em conclusão, a técnica de cone único resultou em menor penetração do cimento do que as outras técnicas, que não diferiram significativamente uma da outra. No entanto, a espessura do cimento foi fortemente dependente da técnica de obturação. O Termafill® demonstrou uma relação superior de GP, seguida por condensação contínua das ondas, condensação lateral e cone único. (AU)

Deficiency of the transcription factor PLAG1 results in aberrant coiling and morphology of the epididymis / 亚洲男科学杂志(英文版)

Mice deficient in the transcription factor pleomorphic adenoma gene 1 (PLAG1) exhibit reproductive issues that are characterized, in part, by decreased progressive sperm motility in the male. However, the underlying cause of this impairment is unknown. As epididymal transit is critical for sperm maturation and motility, the morphology of the epididymis of Plag1-deficient mice was investigated and the spatial expression patterns of PLAG1 protein and mRNA were identified. Using X-gal staining and in situ hybridization, PLAG1 was shown to be widely expressed in both the epithelium and stroma in all regions of the mouse epididymis. Interestingly, the X-gal staining pattern was markedly different in the cauda, where it could be suggestive of PLAG1 secretion into the epididymal lumen. At all ages investigated, the morphology of epididymides from Plag1 knockout (KO) mice was aberrant; the tubule failed to elongate and coil, particularly in the corpus and cauda, and the cauda was malformed, lacking its usual bulbous shape. Moreover, the epididymides from Plag1 KO mice were significantly reduced in size relative to body weight. In 20% of Plag1-deficient mice, the left testicle and epididymis were lacking. The impaired morphogenesis of the epididymal tubule is likely to be a major contributing factor to the fertility problems observed in male Plag1-deficient mice. These results also establish PLAG1 as an important regulator of male reproduction, not only through its involvement in testicular sperm production, but also via its role in the development and function of the epididymis.

Histological evaluation of nephroprotective effect of Solanum nigrum fruit extract against gentamicin induced nephrotoxicity in experimental rats

Background: Drug induced nephrotoxicity, one of the most common renal problem, is a challenge to deal with especially in patients with renal dysfunction. It is responsible for 20% cases of acute renal failure in the community. Modern medicines are costly and have minimal nephroprotection. Solanum nigrum fruit extract, a cheaper drug, have antioxidant property and may help in nephroprotection.Methods: Total 54 rats were randomised in 3 groups named G10, G20 and G30 according to 10, 20 and 30 days of treatment. In each groups, rats were randomly assigned to any of the three subgroups i.e., control C group [received normal saline (2 ml/100 gm/day) orally consecutively for test duration], gentamicin treated (GT) group [received normal saline (2 ml/100 gm/day) orally consecutively for test duration and intraperitoneal gentamicin (40 mg/kg) once daily for last five days] and S. nigrum treated (SNT) group [received S. nigrum orally (200 mg/kg/day) for the test duration and intraperitoneal gentamicin (40 mg/kg) once daily for last five days]. Rats were sacrificed 24 hours after the last dose of gentamicin injection (on 11th, 21st and 31st day). Excised kidneys were weighted and prepared for histological examination.Results: The mean weight of kidneys in GT group was significantly higher than the SNT group in all test durations suggestive of decrease in inflammation in SNT group. This was also reflected histologically as SNT group kidney showed less amount of tubular destruction as compared to GT group.Conclusions: S. nigrum extract provide nephroprotection against gentamicin induced nephrotoxicity.

Preliminary study on clinicopathological features of renal mucinous tubular and spindle cell carcinoma / 肿瘤

Objective: To explore the clinicopathological features and prognosis of mucinous tubular and spindle cell carcinoma (MTSCC) of the kidney in order to improve the understanding of the tumor. Methods: The clinical and pathological information of 15 patients with MTSCC in 7 hospitals were retrospectively collected and analyzed from July 2010 to July 2018. The sections were reviewed by two high-seniority pathologists. The EnVision two-step immunohistochemical staining technique was used to detect the expressions of villin, cytokeratin 7 (CK7), epithelial membrane antigen (EMA), alpha-methylacyl-CoA racemase (AMACR), transducin-like enhancer of split 1 (TLE1), hepatocyte nuclear factor-1p (HNF-1p)and kidney specific calcium binding protein (Ksp-cadherin). The fluorescence in situ hybridization (FISH) was used to detect the synovial sarcoma translocation (SYT)-synovial sarcoma X chromosome breakpoint (SSX) fusion gene in the patients with sarcomatoid changes and positive immunohistochemical staining of TLE1. Finally, the prognostic data of all patients and the relevant literature were reviewed. Results: Among 15 patients with MTSCC, seven of the patients were male and the other eight were female, with an average age of 62 years (ranging from 48 to 75 years). The tumors were found by chance in 12 patients during physical examination, and the other 3 patients developed clinical symptoms such as frequent urine pain or hematuria, including 1 patient with a history of renal calculi for 15 years. The cut surface of tumor is firm and grey or yellow. Except for 2 cases, the majority of tumors were well-circumscribed. Microscopically, 1 case showed neoplastic necrosis, 13 cases showed a mixture of mucinous stroma, tubules and spindle cells, 1 case was mainly composed of spindle cells and mucus, and 1 case was mainly composed of tubule and mucus. Some tumors were with obvious clear cytoplasmic changes, and two cases were accompanied by sarcomatoid differentiation. The immunohistochemical results showed that the positive rates of villin, CK7, EMA, AMACR, TLE1, HNF-1 p and Ksp-cadherin were 20.0% (3/15), 80.0% (12/15), 93.3% (14/15), 80.0% (12/15), 20.0% (3/15), 20.0% (3/15), 93.3% (14/15) and 13.3% (2/15), respectively; the result of FISH excluded synovial sarcoma. Eight patients were followed up wihout other treatment after operation. Bone metastasis occurred in half a year after operation in one patient with follow-up information, while no evidence of local recurrence or distant metastases was identified in the other 7 patients until now. Conclusion: A few cases of MTSCC can metastasize and belong to malignant tumors. The positive expressions of AMACR, CK7 and villin in some cases suggests that the tumor has both proximal and distal renal tubular origins. The positive expression of HNF-ip is correlated with the histological characteristics of MTSCC clear cytoplasm.

Construction of stable cell lines expressing large conductance Ca2+-activated K+ channel α-subunit and molecular mechanism of potassium excretion in this channel / 上海交通大学学报（医学版）

Objective: To construct stable cell lines expressing the large conductance Ca2+-activated K+ channel (MaxiK or BK) α-subunit and to explore the mechanism of potassium excretion via BKα channel. Methods:The BKα plasmid with Myc tag was constructed and transfected into HEK293 cell lines by lipofectamine 2000. The positive monoclonal cell lines were screened by G418, and the expression of BKα was detected by Western blotting and the location of BKα by immunofluorescence. The stable cell lines expressing BKα protein was cultured on slides to form a single cell layer, which was perfused with different potassium ion concentrations of 5 mmol/L and 100 mmol/L, and the single channel patch clamp recorded the ion flux of BKα. Wild type and mutants (G77R, G130R, C140R and R297C) of the inwardly rectifying potassium channel (Kir4.1) were transfected into HEK293 cells stably transfected with BKα, and then the membrane protein was extracted. The expression of BKα was detected by Western blotting. Results:Stable cell lines expressing BKα channel were selected from HEK293 cells after transfection and cellular immunofluorescence verified the expression of BKα channel and its expression on the cell membrane. The channel open frequency (Npo) of BKα increased rapidly when perfused with 100 mmol/L potassium. After being transfected with wild type or mutants of Kir4.1, the membrane expression of BKα in the stable cell lines showed significant difference among these groups (P<0.05). Conclusion:The HEK293 cell lines stably expressing BKα have been successfully constructed. BKα channel can be activated by high potassium solutions. The function of the BKα subunit can be related to Kir4.1 channel, which may be attributed to the depolarization of the cells transfected by Kir4.1 mutants.

Research progress in laser treatment of dentin hypersensitivity / 口腔疾病防治

@#Dentin hypersensitivity is mainly associated with abrasion, wear, acid etching, cracking, wedge-shaped defects, enamel hypoplasia, caries, a lack of neck enamel, and cementum coverage accompanied by gingival recession, resulting in direct exposure of dentin tubules to the oral environment. Dentin hypersensitivity is mainly treated by sealing the exposed dentin tubules and reducing the excitability of the pulpal nerves. Laser therapy, as a safe, fast and convenient treatment measure, has achieved good results both alone and in combination with other medicines and has attracted increasing attention. This paper reviews the research progress in the treatment of dentin hypersensitivity using several common laser therapy mechanisms with parameter selections in various scopes of application. A review of the literature shows that the Nd:YAG laser has a strong penetrating power and a large thermal effect and can denature and coagulate proteins in dentin tubules in a very short time; while the CO2 laser causes little damage to the dental pulp and has an obvious immediate curative effect by high absorption of water molecules in hydroxyapatite and the dentin surface is melted and recrystallized; the Er:YAG laser has high water absorption and a large thermal effect and can block the tubules by evaporating the fluid in the tubules and depositing salts; Er, Cr:YSGG laser energy can be fully absorbed by hydroxyapatite and water and result in desensitization by cutting hard tissues and melting periodontal dentin of the tube simultaneously; The Ga-Al-As semiconductor laser and He:Ne laser are low energy. The laser dosage mainly changes the permeability of nerve fibers to potassium and sodium ions and depolarizes the nerve fibers, producing an analgesic effect. The combination of a laser with a desensitizer can improve the clinical efficacy of treating dentin hypersensitivity.