Porphyromonas gingivalis infection causes umbilical vein endothelial barrier dysfunction in vitro by down-regulating ZO-1, occludin and VE-cadherin expression / 南方医科大学学报

OBJECTIVE@#To explore the molecular mechanisms of Porphyromonas gingivalis infection-induced umbilical vein endothelial barrier dysfunction in vitro.@*METHODS@#Human umbilical vein endothelial cells (HUVECs) were cultured in vitro, and after the formation of the endothelial barrier, the cells were infected with P. gingivals at a multiplicity of infection (MOI). The transepithelial electrical resistance (TEER) of the cell barrier was measured, and FITC-dextran trans-endothelial permeability assay and bacterial translocation assay were performed to assess the endothelial barrier function. The expression levels of cell junction proteins including ZO-1, occludin and VE-cadherin in the cells were examined by qRT-PCR and Western blotting.@*RESULTS@#In freshly seeded HUVECs, TEER increased until reaching the maximum on Day 5 (94 Ωcm2), suggesting the formation of the endothelial barrier. P. gingivals infection caused an increase of the permeability of the endothelial barrier as early as 0.5 h after bacterial inoculation, and the barrier function further exacerbated with time, as shown by significantly lowered TEER, increased permeability of FITC-dextran (40 000/70 000), and increased translocation of SYTO9-E. coli cross the barrier. MTT assay suggested that P. gingivals infection did not significantly affect the proliferation of HUVECs (P>0.05), but in P. gingivalsinfected cells, the expressions of ZO-1, occludin and VE-cadherin increased significantly at 24 and 48 h after bacterial inoculation (P < 0.05).@*CONCLUSION@#P. gingivals may disrupt the endothelial barrier function by down-regulating the expressions of the cell junction proteins (ZO-1, occludin, VE-cadherin) and increasing the permeability of the endothelial barrier.

Curcumin-rich extract from Central Brazil Curcuma longa protects human umbilical vein endothelial cells under oxidative stress / Extrato rico em Curcumina do Brasil Central Curcuma longa protege células endoteliais de veias umbilicais humanas sob estresse oxidativo

INTRODUCTION: In vascular diseases, the interruption of the local blood flow and the subsequent reperfusion of oxygen can cause deleterious oxidative effects on the cells. Turmeric (Curcuma longa L.) presents the capacity to neutralize free radicals along with preventive and therapeutic effects for several diseases. OBJECTIVE: To analyze the bioactive compounds and the antioxidant capacity of the ethanolic extract of Curcuma (EEC), to evaluate its effect on human umbilical vein endothelial cells, and to analyze its effect on cellular signaling pathways. METHODS: Cells were exposed to different concentrations of EEC for 24, 48, and 72 h. Folin-Ciocalteau test, HPLC-Fluorescence analysis, and DPPH method were used to determine the phenolic compounds, curcumin content, and antioxidant action, respectively; the tetrazolium salt reduction to obtain cell viability, cytotoxicity, and the concentration that inhibits 50% of cell viability; and the immunocytochemistry technique to analyze the expression of caspase3, SIRT1, and mTOR. RESULTS: We found the presence of polyphenols in the classes of phenolic acids and curcuminoids in EEC, with 16.7% curcumin content. The number of antioxidants needed to reduce the initial DPPH concentration by 50% was 18.1 µmol/g. The extract mitigated cell damage at a dosage of 100 µg/ml, decreased the immunoexpression of caspase3, and promoted the signaling of the SIRT1 and mTOR survival pathways. CONCLUSION: EEC had a protective effect on human umbilical vein endothelial cells, subjected to oxidative stress, with decreased apoptosis (caspase3) at lower concentrations, cytoprotection by maintaining essential cell functions (mTOR), and signaling of the survival pathway (SIRT1).

INTRODUÇÃO: Em doenças vasculares, a interrupção do fluxo sanguíneo locale subsequente reperfusão de oxigênio pode causar efeitos deletérios e danos irreparáveis às células. Curcuma (Curcuma longa L.) neutraliza radicais livres além de apresentar efeitos preventivos e terapêuticos. OBJETIVO: Caracterizar os compostos bioativos e a capacidade antioxidante do extrato etanólico de cúrcuma (EEC); avaliar seu efeito nas células endoteliais da veia umbilical humana, e analisar a expressão de vias de sinalização celular. MÉTODOS: As células foram expostas a diferentes concentrações de EEC por 24, 48 e 72 horas. Utilizamos o teste de Folin-Ciocalteau, análise por HPLC-Fluorescência e método DPPH para determinar os compostos fenólicos, conteúdo de curcumina e ação antioxidante, respectivamente; o método de redução de tetrazólio para viabilidade celular, a citotoxicidade e a concentração que inibe 50% da viabilidade celular; e a técnica de imunocitoquímica para analisar a expressão de caspase3, SIRT1 e mTOR. RESULTADOS: Observou-se presença de polifenóis nas classes de ácidos fenólicos e curcuminóides no EEC, com teor de curcumina de 16,7%. A quantidade de antioxidante necessária para reduzir a concentração inicial de DPPH em 50% foi de 18,1 µmol/g. O extrato mitigou o dano celular na dosagem de 100 µg/ml, diminuiu a imunoexpressão da caspase3 e promoveu a sinalização das vias de sobrevivência SIRT1 e mTOR. CONCLUSÃO: O EEC teve efeito protetor nas células endoteliais de veia umbilical humana, submetidas ao estresse oxidativo, com diminuição da apoptose (caspase3) em concentrações mais baixas, citoproteção pela manutenção das funções celulares essenciais (mTOR) e sinalização da via de sobrevivência (SIRT1).

Ketamine ameliorates hypoxia-induced endothelial injury in human umbilical vein endothelial cells

OBJECTIVES: Hypoxia leads to endothelial cell inflammation, apoptosis, and damage, which plays an important role in the complications associated with ischemic cardiovascular disease. As an oxidoreductase, p66Shc plays an important role in the regulation of reactive oxygen species (ROS) production and apoptosis. Ketamine is widely used in clinics. This study was designed to assess the potential protective effect of ketamine against hypoxia-induced injury in human umbilical vein endothelial cells (HUVECs). Moreover, we explored the potential mechanism by which ketamine protected against hypoxia-induced endothelial injury. METHODS: The protective effects of ketamine against hypoxia-induced injury was assessed using cell viability and adhesion assays, quantitative polymerase chain reaction, and western blotting. RESULTS: Our data showed that hypoxia reduced HUVEC viability, increased the adhesion between HUVECs and monocytes, and upregulated the expression of endothelial adhesion molecules at the protein and mRNA levels. Moreover, hypoxia increased ROS accumulation and upregulated p66Shc expression. Furthermore, hypoxia downregulated sirt1 expression in HUVECs. Alternatively, ketamine was shown to reverse the hypoxia-mediated reduction of cell viability and increase in the adhesion between HUVECs and monocytes, ameliorate hypoxia-induced ROS accumulation, and suppress p66Shc expression. Moreover, EX527, a sirt1 inhibitor, reversed the protective effects of ketamine against the hypoxia-mediated reduction of cell viability and increase in adhesion between HUVECs and monocytes. CONCLUSION: Ketamine reduces hypoxia-induced p66Shc expression and attenuates ROS accumulation via upregulating sirt1 in HUVECs, thus attenuating hypoxia-induced endothelial cell inflammation and apoptosis.

Absceso hepático neonatal asociado a cateterización de vasos umbilicales / Neonatal liver abscess associated with an umbilical catheter

INTRODUCCIÓN. Los abscesos hepáticos piógenos en neonatos son una entidad rara con menos de cien casos reportados en la literatura mundial. El diagnóstico en esta edad es difícil porque tanto signos y síntomas son inespecíficos y se relacionan con sepsis. OBJETIVOS. Identificar la clínica y características patológicas del absceso hepático en neonatos y correlacionar al posicionamiento de catéter umbilical en hígado como factor de riesgo asociado para desarrollar absceso hepático en neonatos. MATERIALES Y MÉTODOS. Estudio retrospectivo, descriptivo, informes de casos, con una muestra y población conocida, de 5 pacientes recién nacidos. Criterios de Inclusión: neonatos de ambos sexos con edad gestacional mayor a las 34 semanas de gestación, con antecedentes de mal posicionamiento de catéteres umbilicales documentado mediante un examen de imagen "radiografía". Criterios de Exclusión: pacientes sin el antecedente de cateterización umbilical y/o que no se haya documentado mediante estudio de imagen con la evidencia del catéter colocado de manera errónea en tejido hepático. Se utilizó la base de datos registrada en los archivos del Servicio de Gastroenterología Pediátrica de las interconsultas solicitadas del Servicio de Neonatología y Unidad de Cuidados Intensivos Neonatales del, Hospital Civil de Guadalajara "Dr. Juan I. Menchaca". Jalisco- México, en un tiempo de 5 años, desde Enero 2013 a 2018. RESULTADOS. Fueron cinco pre términos (n=5) entre 34 y 36 semanas, con edad gestacional media de 35 semanas y 2039 gr de peso al nacer. El promedio de días para el diagnóstico fue de 7,8. Cuatro pacientes tuvieron hepatomegalia 80,0% (4;5), el volumen de los abscesos fueron entre 2 y 12mm en lóbulo hepático derecho. CONCLUSIÓN. El absceso hepático es poco común pero su clínica debe sospecharse en un recién nacido con sepsis y antecedente de cateterismo umbilical. El manejo y evolución clínico favorable en el 100,0 % (5;5), de los neonatos y no se requirió resolución quirúrgica.

INTRODUCTION. Pyogenic liver abscesses in neonates are a rare entity with less than one hundred cases reported in the world literature. Diagnosis at this age is difficult because both signs and symptoms are nonspecific and are related to sepsis. OBJECTIVES. Identify the clinical and pathological characteristics of liver abscess in neonates and correlate to the positioning of umbilical catheters in the liver as an associated risk factor for developing liver abscess in neonates. MATERIALS AND METHODS. Retrospective, descriptive study, case reports, with a sample and known population, of 5 newborn patients. Inclusion Criteria: neonates of both sexes with gestational age over 34 weeks of gestation, with a history of poor positioning of umbilical catheters documented by an "x-ray" imaging exam. Exclusion Criteria: patients without a history of umbilical catheterization and / or who have not been documented by imaging study with evidence of the catheter incorrectly placed in liver tissue. The database registered in the archives of the Pediatric Gastroenterology Service of the requested interconsultations of the Neonatal and Neonatal Intensive Care Unit of the Civil Hospital of Guadalajara "Dr. Juan I. Menchaca ". Jalisco- Mexico, in a time of 5 years, from January 2013 to 2018. RESULTS. There were five pre terms (n = 5) between 34 and 36 weeks, with an average gestational age of 35 weeks and 2039 grams of birth weight. The average number of days for diagnosis was 7,8. Four patients had 80,0% hepatomegaly (4; 5), the volume of abscesses was between 2 and 12mm in the right hepatic lobe. CONCLUSION. Hepatic abscess is uncommon but its clinic should be suspected in a newborn with sepsis and a history of umbilical catheterization. Management and favorable clinical evolution in 100,0% (5; 5) of infants and no surgical resolution was required.

The types of connections between the portal sinus and main portal vein in foetuses / Tipos de conexiones entre el seno portal y la vena porta principal en fetos

In the prenatal period, the three types of connections between the portal sinus and main portal vein have been published in the literature: T, X and H-shaped. The T type is the most frequent in the literature, and the aim of our study is to define the percentage of the connection types during the prenatal period in our population. In this prospective study, 237 women between 20 and 38 weeks of pregnancy without a foetal anomaly or pregnancy-related complications were included, and the precordial veins of the foetuses were examined using a wide-band color Doppler technique. The types of connections were determined by two specialists according to the shape of the colour coded vessels in Doppler examinations. The criteria of Czubalski & Aleksandrowicz (2000) were used. All of the connection types in patients were confirmed using video clips and were stored in the picture archiving and communication system. In 237 patients, the types of connection were determined by the first specialist as 189 foetuses (79.7 %) with the X-shaped or side-to-side connection, 16 foetuses (6.8 %) with the T-shaped or end-to-side type and 32 foetuses (13.5 %) with the H-shaped or parallel-coursed vessels connected with a short segment. The most common types of connections between the portal sinus and main portal vein in foetuses are X shaped or side-to-side, which is contrary to previous studies.

En el período prenatal, se han publicado en la literatura los tres tipos de conexiones entre el seno portal y la vena porta principal: en forma de T, X y H. El tipo T es el más frecuente, y el objetivo de nuestro estudio fue definir el porcentaje de tipos de conexión durante el período prenatal en nuestra población. En este estudio prospectivo, se incluyeron 237 mujeres entre 20 y 38 semanas de embarazo, sin anomalías fetales o complicaciones relacionadas con el embarazo, y se examinaron las venas precordiales de los fetos utilizando una técnica Doppler de banda ancha. Los tipos de conexiones fueron determinados por dos especialistas según la forma de los vasos codificados por color en los exámenes Doppler. Se utilizaron los criterios del estudio de Czubalski & Aleksandrowicz. Todos los tipos de conexión en los pacientes se confirmaron mediante videoclips y se almacenaron en el sistema de comunicación y en archivo de imágenes. En 237 pacientes, el primer especialista determinó en 189 fetos (79,7 %) la conexión en forma de X o de lado a lado; en 16 fetos (6,8 %) la forma de T o Tipo de extremo a lado; y en 32 fetos (13,5 %) los vasos en forma de H o paralelos, conectados con un segmento corto. Los tipos más comunes de conexiones entre el seno portal y la vena porta principal en los fetos son en forma de X o de lado a lado, lo que es contrario a estudios anteriores.

Inhibiting miR-186 expression alleviates mitochondrial damage in hypoxic human umbilical vein endothelial cells / 南方医科大学学报

OBJECTIVE@#To investigate the effect of miR-186 inhibition on the expression of hypoxia-inducible factor-1α (HIF-α) and mitochondrial function in hypoxic vascular endothelial cells.@*METHODS@#Human umbilical vein endothelial cells (HUVECs) cultured in routine or hypoxic conditions for 6 h were examined for the expression of miR-186. A miR-186 inhibitor was transfected in the HUVECs, and the cells were subsequently cultured in hypoxic condition for 6 h to observe the changes in the mitochondrial structure under an electron microscope. The changes in the mRNA and protein expressions of HIF-1α in response to miR-186 interference were tested using real-time fluorescent quantitative PCR and Western blotting.@*RESULTS@#The expression of miR-18 was mildly increased in HUVECs after hypoxic exposure for 6 h (=0.0188). Interference of miR-186 expression obviously promoted the mRNA and protein expressions of HIF-1α in HUVECs. In hypoxic conditions, miR-186 interference significantly reduced mitochondrial damage in HUVECs as observed under electron microscope (=0.0297).@*CONCLUSIONS@#Inhibition of miR-186 protects vascular endothelial cells against hypoxic injuries by promoting HIF-α expression to lessen mitochondrial damage, suggesting the possibility of targeted miR-186 interference for treatment of hemorrhagic shock.

Docosahexaenoic acid reduces adenosine triphosphate-induced calcium influx via inhibition of store-operated calcium channels and enhances baseline endothelial nitric oxide synthase phosphorylation in human endothelial cells

Docosahexaenoic acid (DHA), an omega-3-fatty acid, modulates multiple cellular functions. In this study, we addressed the effects of DHA on human umbilical vein endothelial cell calcium transient and endothelial nitric oxide synthase (eNOS) phosphorylation under control and adenosine triphosphate (ATP, 100 µM) stimulated conditions. Cells were treated for 48 h with DHA concentrations from 3 to 50 µM. Calcium transient was measured using the fluorescent dye Fura-2-AM and eNOS phosphorylation was addressed by western blot. DHA dose-dependently reduced the ATP stimulated Ca²⁺-transient. This effect was preserved in the presence of BAPTA (10 and 20 µM) which chelated the intracellular calcium, but eliminated after withdrawal of extracellular calcium, application of 2-aminoethoxy-diphenylborane (75 µM) to inhibit store-operated calcium channel or thapsigargin (2 µM) to delete calcium store. In addition, DHA (12 µM) increased ser1177/thr495 phosphorylation of eNOS under baseline conditions but had no significant effect on this ratio under conditions of ATP stimulation. In conclusion, DHA dose-dependently inhibited the ATP-induced calcium transient, probably via store-operated calcium channels. Furthermore, DHA changed eNOS phosphorylation suggesting activation of the enzyme. Hence, DHA may shift the regulation of eNOS away from a Ca²⁺ activated mode to a preferentially controlled phosphorylation mode.

Portal Hypertension of a Delayed Onset Following Liver Abscesses in a 12-Month-Old Infant: A Case Report and Review of the Literature / 대한소아소화기영양학회지

We report a 12-month-old female infant who had a history of neonatal sepsis with liver micro-abscesses that resolved with intravenous antibiotics during neonatal period. During her neonatal admission period, no umbilical vein catheter was inserted. Also, she did not undergo any abdominal surgeries or had a postnatal history of necrotizing enterocolitis. However, the child developed upper gastrointestinal bleeding in form of hematemesis and melena secondary to esophageal varices at the age of 12 months with an extra-hepatic portal vein obstruction with cavernous transformation and portal hypertension subsequently. The child underwent a successful endoscopic injection sclerotherapy. She is now 20-month-old and has portal hypertension but otherwise asymptomatic. We are proposing the possibility of a delayed-onset portal hypertension as a complication of liver abscess and neonatal sepsis.

Aspectos ultrassonográficos dos componentes umbilicais de bezerros da raça Holandesa durante o processo de involução fisiológica / Ultrasonographic aspects of umbilical components of Holstein calves during the physiological involution process

Afecções umbilicais são comumente encontradas nos bezerros durante o período neonatal, podendo ocasionar graves complicações. Seu diagnóstico muitas vezes não é preciso pela palpação abdominal, sendo a ultrassonografia um valioso exame complementar, pois permite precisão na localização e na extensão das onfalopatias intra-abdominais. Diante disso e da raridade de pesquisas com estabelecimento de padrões ultrassonográficos do umbigo, o presente estudo propôs padronizar os aspectos das imagens ultrassonográficas dos componentes umbilicais em decorrência de sua involução. Foram avaliados 23 bezerros Holandeses, do nascimento até os 30 dias de vida, em cuja região umbilical se usou como antisséptico tintura de iodo em diferentes concentrações. Os resultados evidenciaram que veia e artérias umbilicais perdem suas características de vasos, assumindo aspecto de ligamento por proliferação de tecido fibroso. Nesse processo, o tecido fibroso, inicialmente presente na região interna da parede do vaso, segue, com a involução, em direção à luz, sendo observado mais precocemente em porções dos vasos mais distantes do umbigo externo, não havendo distinção de comportamento determinada pela antissepsia. Para aproveitamento do exame ultrassonográfico, é importante o conhecimento dos aspectos das imagens durante o processo de involução dos componentes umbilicais, de acordo com cada fase, sendo assim possível o diagnóstico das diferentes alterações nessas estruturas.(AU)

Umbilical diseases of calves happen during neonatal period and may lead to severe complications. The diagnosis is usually through abdominal palpation although it is not very accurate, thus ultrasound provides a valuable complementary exam to establish a precise diagnosis of location and extent of intra-abdominal umbilical diseases. Given those facts and the lack of established standards for umbilical ultrasound imaging the present study proposal was to standardize the physiological aspects of umbilical components during involution. Ultrasound images were obtained for 23 Holstein calves, from birth until 30 days of life. Iodine tincture of different concentrations was used for umbilical region antisepsis. Results show that umbilical vein and arteries lose their vessel characteristics, becoming similar to ligament, due to the proliferation of fibrous tissue. The growth pattern of the fibrous tissue was from the vessel walls growing toward vascular lumen. The involution process begins at the most distant part and did not vary with antiseptic concentrations. To obtain a reliable ultrasound exam it's important to know the aspects of imaging patterns according to each phase of umbilical involution, thus leading to an accurate diagnosis of structural variations and umbilical diseases.(AU)

Persistent right umbilical vein: a study using serial sections of human embryos and fetuses / 대한해부학회지

Persistent right umbilical vein (PRUV) is a common anomaly of the venous system. Although candidates for future PRUV were expected to occur more frequently in earlier specimens, evaluation of serial horizontal sections from 58 embryos and fetuses of gestational age 5–7 weeks found that only two of these embryos and fetuses were candidates for anomalies. In a specimen, a degenerating right umbilical vein (UV) joined the thick left UV in a narrow peritoneal space between the liver and abdominal cavity, and in the other specimen, a degenerating left UV joined a thick right UV in the abdominal wall near the liver. In these two specimens, the UV drained into the normal, umbilical portion of the left liver. These results strongly suggested that, other than the usual PRUV draining into the right liver, another type of PRUV was likely to consist of the right UV draining into the left liver.

Adiponectin is protective against endoplasmic reticulum stress-induced apoptosis of endothelial cells in sepsis

Endoplasmic reticulum (ER) stress is a critical molecular mechanism involved in the pathogenesis of sepsis. Hence, strategies for alleviating this stress may be essential for preventing cardiovascular injuries under sepsis. Adiponectin is secreted by adipocytes and its levels are decreased in sepsis. The purpose of this study was to investigate the protective effects of adiponectin treatment on endothelial cells and its mechanism. Male Wistar rats underwent cecal ligation and puncture (CLP) before being treated with adiponectin (72 and 120 μg/kg). The levels of malondialdehyde (MDA) in plasma, histological structure, and apoptosis of endothelial cells were evaluated. In vitro, human umbilical vein endothelial cells (HUVECs) were treated with adiponectin at 10 and 20 μg/mL for 24 h after stimulation by lipopolysaccharide (LPS). The levels of reactive oxygen species (ROS), ultrastructure, rate of apoptosis, the expression of inositol-requiring enzyme 1α (IRE1α) protein, and its downstream molecules (78 kDa glucose-regulated protein (GRP78), C/EBP homologous protein (CHOP), and caspase-12) were detected. The results showed that the levels of MDA and ROS induced by CLP or LPS stimulation were increased. Furthermore, endothelial cell apoptosis was increased under sepsis. The IRE1α pathway was initiated, as evidenced by activated IRE1α, increased GRP78, and up-regulated CHOP and caspase-12 in HUVECs. Following treatment with adiponectin, the number of apoptotic endothelial cells was markedly decreased. These findings demonstrated that treatment with adiponectin decreased apoptosis of endothelial cells caused by sepsis by attenuating the ER stress IRE1α pathway activated by oxidative stress.

Accuracy of chest radiography for positioning of the umbilical venous catheter / Acurácia da radiografia de tórax para o posicionamento do cateter venoso umbilical

Abstract Objectives: To evaluate the accuracy of the simultaneous analysis of three radiographic anatomical landmarks - diaphragm, cardiac silhouette, and vertebral bodies - in determining the position of the umbilical venous catheter distal end using echocardiography as a reference standard. Methods: This was a cross-sectional, observational study, with the prospective inclusion of data from all neonates born in a public reference hospital, between April 2012 and September 2013, submitted to umbilical venous catheter insertion as part of their medical care. The position of the catheter distal end, determined by the simultaneous analysis of three radiographic anatomical landmarks, was compared with the anatomical position obtained by echocardiography; sensitivity, specificity, positive predictive value, negative predictive value, and accuracy were calculated. Results: Of the 162 newborns assessed by echocardiography, only 44 (27.16%) had the catheter in optimal position, in the thoracic portion of the inferior vena cava or at the junction of the inferior vena cava with the right atrium. The catheters were located in the left atrium and interatrial septum in 54 (33.33%) newborns, in the right atrium in 26 (16.05%), intra-hepatic in 37 (22.84%), and intra-aortic in-one newborn (0.62%). The sensitivity, specificity and accuracy of the radiography to detect the catheter in the target area were 56%, 71%, and 67.28%, respectively. Conclusion: Anteroposterior radiography of the chest alone is not able to safely define the umbilical venous catheter position. Echocardiography allows direct visualization of the catheter tip in relation to vascular structures and, whenever possible, should be considered to identify the location of the umbilical venous catheter.

Resumo Objetivos: Avaliar a acurácia da análise simultânea dos três marcos anatômicos radiográficos - diafragma, silhueta cardíaca e corpos vertebrais - na determinação da posição da extremidade distal do cateter venoso umbilical com a ecocardiografia como padrão de referência. Métodos: Estudo transversal, observacional, com inclusão prospectiva de dados de todos os neonatos nascidos em uma maternidade pública de referência, entre abril de 2012 e setembro de 2013, submetidos à inserção de cateter venoso umbilical como parte do atendimento clínico. A posição da extremidade distal do cateter, determinada pela análise simultânea dos três marcos anatômicos radiográficos, foi comparada com a posição anatômica obtida pela ecocardiografia. Sensibilidade, especificidade, valor preditivo positivo, valor preditivo negativo e acurácia foram calculados. Resultados: Dos 162 recém-nascidos avaliados por ecocardiografia, somente 44 (27,16%) estavam com o cateter em posição ótima, na porção torácica da veia cava inferior ou na junção da veia cava inferior com o átrio direito. Os cateteres foram localizados no átrio esquerdo e no septo interatrial em 54 (33,33%), no átrio direito em 26 (16,05%), no intra-hepático em 37 (22,84%) e na aorta em um recém-nascido (0,62%). A sensibilidade, especificidade e acurácia da radiografia para detectar cateter na zona-alvo foram de 56%, 71% e 67,28%, respectivamente. Conclusão: A radiografia anteroposterior de tórax isolada não é capaz de definir com segurança a posição do cateter venoso umbilical. A ecocardiografia permite a visibilização direta da ponta do cateter em relação às estruturas vasculares e, sempre que possível, deve ser considerada para localização do cateter venoso umbilical.

Studying the Effect of Downregulating Autophagy-Related Gene LC3 on TLR3 Apoptotic Pathway Mediated by dsRNA in Hepatocellular Carcinoma Cells / Journal of the Korean Cancer Association, 대한암학회지

PURPOSE: The purpose of this study is to examine the role of the double-stranded RNA (dsRNA) activated Toll–interleukin-1 receptor domain-containing adaptor inducing interferon β (TRIF) signal pathway in triggering apoptosis in hepatocellular carcinoma (HCC) cells. MATERIALS AND METHODS: First, siRNA targeted autophagy–related gene LC3 (pU6H1-LC3 siRNA and siLC3) and a dsRNA used as a Toll-like receptor 3 (TLR3) ligand was constructed and synthesized, respectively. Then, a human HCC cell line was transfected with dsRNA, siLC3, and cotransfected with siLC3 and dsRNA (siLC3+dsRNA), respectively. Finally, quantification real-time polymerase chain reaction, western blotting, and immunofluorescence staining were used in the HCC line (SMMC7721), and MTT assay, flow cytometry, terminal deoxynucleotidyl transferase-mediated dUTP nick-end-labeling, and transmission electron microscopy were used in an HCC xenograft model of nude mice. Human umbilical vein endothelial cell tube forming assay, color Doppler ultrasonographic flow image examination, and CD34-positive microvessel density were used in vitro and in vivo. RESULTS: Compared with untreated cells, the protein and mRNA expression of TLR3 and TRIF was up-regulated, in order, siLC3+dsRNA, dsRNA, and siLC3. Expression of LC3 was obviously down-regulated and the autophagosomes were significantly decreased in siLC3+dsRNA and siLC3, whereas in dsRNA (p < 0.05). LC3 and TRIF colocation was observed in HepG2 cells. Decreased cell viability, increased apoptosis, decrease in xenograft tumor volume, and angiogenesis potential were also observed in order (p < 0.05). CONCLUSION: Suppression of intracellular autophagy resulted in decreased degradation of TRIF protein, which can promote triggering of apoptosis by the TLR3-TRIF pathway. dsRNA and siLC3 could play anticancer roles in coordination.

Rol de los marcadores ecográficos secundarios de primer trimestre para la detección de trisomía 21 / Role of first trimester secondary ultrasound markers for trisomy 21 detection

OBJETIVO: Analizar los resultados de los marcadores ecográficos secundarios (hueso nasal, onda a del ductus venoso y regurgitación tricuspídea) y valorar su efectividad para la detección de trisomía 21 y su utilidad para la reducción del número de pruebas invasivas. MÉTODOS: Tras la realización del test combinado de primer trimestre a toda paciente con un riesgo entre 1/101-1/1000 se realizó la valoración de los marcadores secundarios. RESULTADOS: Desde Enero de 2014 a Mayo de 2015 se realizaron 2.660 test combinados del primer trimestre valorándose la edad materna, la traslucencia nucal y la PAPP-A y ßhCG, teniendo una sensibilidad del 90% y una tasa de falsos positivos del 3,2%. Hubo 10 fetos con trisomía 21. La sensibilidad de hueso nasal, ductus venoso y regurgitación tricuspídea fue del 22,2%, 50% y 50% y la especificidad del 99,8%, 96,9% y 98,8% respectivamente. La sensibilidad global del test contingente fue del 90%, con una reducción de la tasa de falsos positivos al 1,6%, lo que se reduciría de 171 a 148 el número de amniocentesis. CONCLUSIÓN: El test contingente es una buena herramienta para reducir la tasa de falsos positivos respecto al test combinado sin disminuir la tasa de detección y con ello reducir la tasa de pruebas invasivas.

AIMS: To analyze the results of the secondary sonographic markers (nasal bone, wave ductus venosus and tricuspid regurgitation) and evaluate its effectiveness for the detection of trisomy 21 and thus reduce the number of invasive tests. METHODS: After completing the first trimester combined test, all patients with a risk between 1/101-1/1000 were evaluated the secondary sonographic markers. RESULTS: From January 2014 to May 2015 2660 combined test being assessed maternal age, nuchal translucency and PAPP-A and ßhCG were performed, with a sensitivity of 90% and a false positive rate of 3.2%. 10 fetuses with trisomy 21 were observed. The sensitivity of nasal bone, ductus venosus and tricuspid regurgitation was 22.2%, 50% and 50% and specificity was 99.8%, 96.9% and 98.8% respectively. The overall sensitivity of contingent test was 90%, with a reduction in false positive rate to 1.6%, which would decrease the number of amniocentesis from 171 to 148. CONCLUSION: The contingent test is a good tool to reduce the rate of false positives with respect to the combined test without decreasing the detection rate and thereby reduce the rate of invasive testing.

The Association between N-terminal Pro-Brain Natriuretic Peptide Levels in the Umbilical Vein and Amniotic Fluid Volume Abnormalities / Associação entre níveis de peptídeo natriurético prócerebral N-terminal na veia umbilical e as anormalidades do volume de líquido amniótico

Abstract Purpose The amniotic fluid volume (AFV) is known as a predictor for the wellness of a fetus. We aimed to investigate whether N-terminal pro-brain natriuretic peptide (NTproBNP) levels reflect AFV abnormalities in otherwise normal fetuses. Methods We recruited 24 women with isolated oligohydramnios, 23 women with isolated polyhydramnios, and 36 women with normal AFV at a tertiary referral center. NT-proBNP levels in umbilical venous samples and the individual characteristics of the three groups were compared. One-way ANOVA and Kruskal-Wallis analysis of variance were used for multi-group comparisons of continuous variables. When a significant difference was detected, the Scheffe test was performed as a post-hoc analysis. Proportions were compared using the Chi-square (2) test. Results Maternal age, body mass indices, weight gained in pregnancy and NT-proBNP levels were similar among the three groups. Apgar scores at 1 and 5 minutes significantly correlated with NT-proBNP levels in all newborns (Spearman's r = 0.23 ; p = 0.03 and Spearman's r = 0.24; p = 0.02, respectively). The umbilical venous NTproBNP levels did not differ between newborns who needed mechanical ventilation and those who didn't (p = 0.595). Conclusions NT-proBNP is a biomolecule that may provide insights into the pathogenesis of fetal circulatory problems and subsequent renal failure. Further investigations are warranted.

Resumo Objetivo Investigar se os níveis de peptídeo natriurético pró-cerebral N-terminal (NTproBNP) refletem anormalidades no volume de líquido amniótico (VLA) em fetos normais. Métodos Reunimos 24 mulheres com oligoidrâmnios isolados, 23 com poli-hidrâmnios isolados, e 36 com VLA normal em um centro de referência. Comparamos os níveis de NT-proBNP em amostras venosas umbilicais e características individuais em três grupos. Usamos análise de variância simples (One-way ANOVA) e a análise de variação Kruskal-Wallis para comparação de variáveis contínuas em múltiplos grupos. Quando identificada uma diferença significativa, o teste de Scheffe foi aplicado como uma análise post-hoc. Comparamos proporções usando o teste Qui-quadrado (2). Resultados Idade fértil, índice de massa corporal, ganho de peso na gestação e níveis de NT-proBNP foram similares nos três grupos. Apgar em 1 e 5 minutos correlacionaram significativamente com os níveis de NT-proBNP em todos os recém-nascidos (Spearman's r = 0,23; p = 0,03 e Spearman's r = 0,24; p = 0,02, respectivamente). Os níves de NT-proBNP venoso umbilical não se distinguiram entre os recém-nascidos que precisaram de ventilação mecânica e aqueles que não precisaram (p = 0,595). Conclusões NT-proBNP é um candidato biomolecular que pode contribuir na patogênese de problemas circulatórios fetais e subsequente insuficiência renal. São necessárias futuras investigações.

Effects of Danhong Injection (丹红注射液) and its main components on anticoagulation and fibrinolysis in cultured vein endothelial cells / 中国结合医学杂志

<p><b>OBJECTIVE</b>To observe the effects of Danhong Injection (丹红注射液) and its main components, including daiclzein and hydroxysafflor yellow A (HSYA), on the anticoagulation, fibrinolysis, anti-apoptosis in hypoxia model of vein endothelial cells (VECs).</p><p><b>METHODS</b>VECs were prepared and were put in a hypoxia environment, which consisted of mixed gas of 95% N and 5% CO mixed gas, when reached confluent culture. Five groups used different treatments, including normal control group, hypoxia group, daiclzein group, HSYA group and Danhong Injection group. The VECs were identified by fluorescence double labeling methods. The morphology was observed by a phase contrast microscopy. The effects of Danhong Injection, daiclzein and HSYA on 6 keto prostaglandin F1α (6-keto-PGF1α) level was measured by the method of radioimmunoassay (RIA). Superoxide dismutase (SOD) activity was tested by water soluble tetrazolium salt. The content of malondialdehyde (MDA) was measured by thiobarbituric acid. The activities of tissue-type plasminogen activator (t-PA) and plasminogen activator inhibitor (PAI) were measured by the method of chromogenic substrate. The contents of endothelin (ET) and nitric oxide (NO) were detected by non-equilibrium RIA and enzymelinked immunosorbent assay. Cells apoptosis rate was determined by flow cytometry.</p><p><b>RESULTS</b>Compared with the normal control group, the floating cells number, PAI activity, ET and MDA contents, and cells apoptosis rate in the culture solution of hypoxia group were all significantly increased, whereas the 6-keto-PGF1α and NO contents, and t-PA and SOD activities were decreased significantly (P<0.01). Compared with the hypoxia group, Danhong Injection markedly increased the 6-keto-PGF1α content and SOD activity, regulated PAI and t-PA activities, ET and NO contents, and decreased MDA content and cells apoptosis rate (P<0.05 or P<0.01).</p><p><b>CONCLUSIONS</b>Danhong Injection and its main components played an important role in protecting primary VECs from hypoxic damage by regulating the secretion and vasomotor function of VECs. The function of Danhong Injection was most remarkable.</p>

Y-27632 reduces the MMP2 and MMP9 expression in endothelial cell via inhibition of ROCK signal pathway / 中南大学学报(医学版)

OBJECTIVE@#To explore the effect of ROCK inhibitor Y-27632 on the matrix metalloproteinase 2 and 9 (MMP2 and MMP9) gene expression and activity in tumor necrosis factor α (TNF-α)-treated human umbilical vein endothelial cell (HUVEC).
@*METHODS@#HHUVEC was divided into 3 groups, a control group, a TNF-α group, and a TNF-α plus Y-27632 group. The expressions of vascular cell adhesion molecule-1 (VCAM-1), intercellular adhesion molecule-1 (ICAM-1), MMP2 and MMP9 were examined by real-time PCR. The MMP2/9 activity was measured by gelatin zymography.
@*RESULTS@#Compared to the control group, the mRNA expressions of ICAM-1, VCAM-1, MMP2 and MMP9 were increased TNF-α-treated cells, which were suppressed by ROCK inhibitor (P<0.01). The MMP2/9 activity was elevated in TNF-α-treated cells, which was reversed by ROCK inhibitor (P<0.05).
@*CONCLUSION@#ROCK inhibitor can suppress TNF-α-induced inflammation in endothelial cells through down-regulation of MMP2/9.

Restricted Usage of Prophylactic Antibiotics in Extremely Low Birth Weight Infants for Early-Onset Sepsis as Quality Improvement Program

PURPOSE: The prevalence of antibiotics resistant bacterial infection among preterm infants has been increased due to indeliberate use of prophylactic broad spectrum antibiotics. The objective of this study was to assess the effectiveness of restricted usage of prophylactic antibiotics by comparing the incidence of culture proven early onset sepsis (EOS). METHODS: This was a retrospective cohort study for extremely low birth weight infants who were born in Seoul National University Children's Hospital during 2009-2014. The groups were divided into two periods, from 2009 to 2011 (period I) and from 2012 to 2014 (period II) based on the implementation on quality improvement activity since 2012. The indication of prophylactic antibiotics were; 1) umbilical vein catheter (UVC) insertion for resuscitation in delivery room, 2) prolonged preterm premature rupture of membrane >18 hours, 3) maternal fever during labor or sustained septic amniotic fluid. The incidence of EOS and the rate of empirical antibiotics usage were compared between two periods. RESULTS: A total of 245 infants were admitted to the neonatal intensive care unit during the study period. Baseline demographics and clinical characteristics were similar between two periods except UVC insertion rate. The rate of empirical antibiotics usage significantly decreased in period II (71.1% for period I vs. 56.4% for period 2, P=0.022). Incidence of EOS was not different between two periods whether prophylactic antibiotics use or not. CONCLUSION: Quality improvement for reducing prophylactic antibiotics use may be effective to reduce a use of antibiotics without increasing EOS.

Aspirin-Triggered Resolvin D1 Inhibits TGF-β1-Induced EndMT through Increasing the Expression of Smad7 and Is Closely Related to Oxidative Stress

The endothelial-mesenchymal transition (EndMT) is known to be involved in the transformation of vascular endothelial cells to mesenchymal cells. EndMT has been confirmed that occur in various pathologic conditions. Transforming growth factor β1 (TGF-β1) is a potent stimulator of the vascular endothelial to mesenchymal transition (EMT). Aspirin-triggered resolvin D1 (ATRvD1) has been known to be involved in the resolution of inflammation, but whether it has effects on TGF-β1-induced EndMT is not yet clear. Therefore, we investigated the effects of AT-RvD1 on the EndMT of human umbilical vein vascular endothelial cells line (HUVECs). Treatment with TGF-β1 reduced the expression of Nrf2 and enhanced the level of F-actin, which is associated with paracellular permeability. The expression of endothelial marker VE-cadherin in HUVEC cells was reduced, and the expression of mesenchymal marker vimentin was enhanced. AT-RvD1 restored the expression of Nrf2 and vimentin and enhanced the expression of VE-cadherin. AT-RvD1 did also affect the migration of HUVEC cells. Inhibitory κB kinase 16 (IKK 16), which is known to inhibit the NF-κB pathway, had an ability to increase the expression of Nrf2 and was associated with the inhibition effect of AT-RvD1 on TGF-β1-induced EndMT, but it had no effect on TGF-β1-induced EndMT alone. Smad7, which is a key regulator of TGF-β/Smads signaling by negative feedback loops, was significantly increased with the treatment of AT-RvD1. These results suggest the possibility that AT-RvD1 suppresses the TGF-β1-induced EndMT through increasing the expression of Smad7 and is closely related to oxidative stress.

Multidetector computed tomography in the preoperative staging of gastric adenocarcinoma / Tomografia computadorizada de múltiplos detectores no estadiamento pré-operatório do adenocarcinoma gástrico

Objective: To evaluate the role of multidetector computed tomography in the preoperative investigation of tumor invasion depth and lymph node and metastatic involvement according to the TNM classification, in patients with gastric adenocarcinoma. Materials and Methods: Fifty-four patients with biopsy-confirmed gastric cancer underwent preoperative staging with 64-channel multidetector computed tomography. Two independent radiologists analyzed the images and classified the findings. Sensitivity, specificity, accuracy and overall accuracy were calculated for each observer. The interobserver agreement was also evaluated. Results: The accuracy in the classification of categories T ranged from 74% to 96% for observer 1 and from 80% to 92% for observer 2. The overall accuracy was 70% for both observers. The weighted kappa index was 0.75, consistent with a significant interobserver agreement. The accuracy in the classification of lymph node involvement (category N) ranged from 55% to 79% for observer 1 and from 73% to 82% for observer 2. The evaluation of metastatic involvement showed an overall accuracy of 89.6% for both observers. Conclusion: 64-channel multidetector computed tomography demonstrated clinically relevant accuracy in the preoperative staging of gastric adenocarcinoma as regards invasion depth (T category) and metastatic involvement (M category). .

Objetivo: Avaliar a tomografia computadorizada com múltiplas fileiras de detectores na análise pré-operatória da profundidade de invasão tumoral, acometimento linfonodal e metastático, de acordo com a classificação TNM, em pacientes com adenocarcinoma gástrico. Materiais e Métodos: Cinquenta e quatro pacientes com câncer gástrico foram submetido a estadiamento pré-operatório com tomografia computadorizada de 64 canais de detectores. Dois radiologistas analisaram, independentemente, as imagens e classificaram os achados. A sensibilidade, especificidade, acurácia e acurácia global para cada avaliador foram calculadas. A concordância interobservador também foi avaliada. Resultados: A acurácia na classificação das categorias T variou entre 74% e 96% para o observador 1 e entre 80% e 92% para o observador 2. A acurácia global foi 70% para ambos os observadores. O kappa ponderado foi 0,75, consistente com uma concordância interobservador substancial. A acurácia na classificação do acometimento linfonodal (categoria N) variou entre 55% e 79% para o observador 1 e entre 73% e 82% para o observador 2. A avaliação do acometimento metastático mostrou acurácia global de 89,6% para ambos os observadores. Conclusão: A tomografia computadorizada com 64 canais de detectores demonstrou acurácia clinicamente relevante no estadiamento pré-operatório do adenocarcinoma gástrico em relação à profundidade de invasão e acometimento metastático. .