Interpretation of standards on training and evaluation of urine analyzers in grass-roots medical unit / 医疗卫生装备

Objective To analyze the training and evaluation standards on urine analyzers in the grass-roots medical unit.Methods The applicability,requirements,subject setup and items of the standards as well as the requirements of evaluation scoring chart were expounded in detail,and the related elementary knowledge was analyzed including the structure,working principle,installation and adjustment,operation,application,maintenance and etc of the analyzer.Combined personnel with the equipment was suggested to ensure the accuracy and high efficiency of the test result.Results The interpretation of the standards contributed to enhancing the quality of medical equipment training and the ability of practical support,and facilitated the examiner to perform annual evaluation with the scoring chart.Conclusion It is necessary to grasp accurately the connotation of the standards,to improve the quality of medical equipment training and to enhance combat support capability for the grass-roots medical unit and examiner.

Evaluation of a fungal collection as certified reference material producer and as a biological resource center

Abstract Considering the absence of standards for culture collections and more specifically for biological resource centers in the world, in addition to the absence of certified biological material in Brazil, this study aimed to evaluate a Fungal Collection from Fiocruz, as a producer of certified reference material and as Biological Resource Center (BRC). For this evaluation, a checklist based on the requirements of ABNT ISO GUIA34:2012 correlated with the ABNT NBR ISO/IEC17025:2005, was designed and applied. Complementing the implementation of the checklist, an internal audit was performed. An evaluation of this Collection as a BRC was also conducted following the requirements of the NIT-DICLA-061, the Brazilian internal standard from Inmetro, based on ABNT NBR ISO/IEC 17025:2005, ABNT ISO GUIA 34:2012 and OECD Best Practice Guidelines for BRCs. This was the first time that the NIT DICLA-061 was applied in a culture collection during an internal audit. The assessments enabled the proposal for the adequacy of this Collection to assure the implementation of the management system for their future accreditation by Inmetro as a certified reference material producer as well as its future accreditation as a Biological Resource Center according to the NIT-DICLA-061.

Effects of berberine and cinnamic acid on palmitic acid-induced intracellular triglyceride accumulation in NIT-1 pancreatic β cells / 中国结合医学杂志

<p><b>OBJECTIVE</b>To investigate the effects of berberine (BBR) and cinnamic acid (CA), the main active components in Jiaotai Pill (, JTP), on palmitic acid (PA)-induced intracellular triglyceride (TG) accumulation in NIT-1 pancreatic β cells.</p><p><b>METHODS</b>Cells were incubated in culture medium containing PA (0.25 mmol/L) for 24 h. Then treatments with BBR (10 μmol/L), CA (100 μmol/L) and the combination of BBR and CA (BBR+CA) were performed respectively. Intracellular lipid accumulation was assessed by Oil Red O staining and TG content was measured by colorimetric assay. The expression of adenosine monophosphate-activated protein kinase (AMPK) protein and its downstream lipogenic and fatty acid oxidation genes, including fatty acid synthase (FAS), acetyl-coA carboxylase (ACC), phosphorylation acetyl-coA carboxylase (pACC), carnitine acyl transferase 1 (CPT-1) and sterol regulating element binding protein 1c (SREBP-1c) were determined by Western blot or real time polymerase chain reaction.</p><p><b>RESULTS</b>PA induced an obvious lipid accumulation and a significant increase in intracellular TG content in NIT-1 cells. PA also induced a remarkable decrease in AMPK protein expression and its downstream targets such as pACC and CPT-1. Meanwhile, AMPK downstream lipogenic genes including SREBP-1c mRNA, FAS and ACC protein expressions were increased. Treatments with BBR and BBR+CA, superior to CA, significantly reversed the above genes changes in NIT-1 pancreatic β cells. However, the synergistic effect of BBR and CA on intracellular TG content was not observed in the present study.</p><p><b>CONCLUSION</b>It can be concluded that in vitro, BBR and BBR+CA could inhibit PA-induced lipid accumulation by decreasing lipogenesis and increasing lipid oxidation in NIT-1 pancreatic β cells.</p>

Estudio epidemiológico de la pediculosis en escuelas básicas del extremo norte de Chile / Epidemiological study of pediculosis in elementary schools of Arica, northern Chile

Introducción: La Pediculosis es una ectoparasitosis que ha recibido poca atención en el norte de Chile, a pesar de que es recurrente en la población infantil escolar. Nuestro objetivo fue determinar la prevalencia de Pediculus humanus capitis, en la población escolar básica de la Provincia de Arica. Pacientes y Método: Se estudió un grupo de 665 alumnos de enseñanza básica del sistema municipalizado, de la Provincia de Arica, durante agosto a noviembre de 2010. Se seleccionaron 467 niños que cumplían con los criterios de inclusión. Previo consentimiento informado de los alumnos y apoderados, el diagnóstico se llevó a cabo a través de la revisión a ojo descubierto de la cabeza de los escolares participantes, en dependencias de cada establecimiento, registrando la presencia de Pediculus humanus capitis, grado de infestación y estadio del ectoparásito. Resultados: De la población analizada, el 40,3 por ciento (188/467) presentó Pediculosis, afectando a las seis escuelas estudiadas. Las mujeres presentaron Pediculosis en un 55,2 por ciento (138/250) y los hombres un 23 por ciento (50/217). Conclusión: La población escolar de la Provincia de Arica está significativamente afectada por Pediculosis y su prevalencia es similar a otras regiones del país.

Pediculosis is an ectoparasitosis infestation that has not received much attention in northern Chile despite it is a common reoccurring condition among school-age children. The objective of this study is to determine the prevalence of Pediculus humanus capitis infestation in elementary schools of Arica. Patients and Method: A group of 665 elementary school students from the Province of Arica were screened between August and November 2010; 467 children who met the inclusion criteria were selected. After parents and students signed an informed consent, the diagnosis was carried out through examination of the head with naked eye in each participating school, recording the presence of Pediculus humanus capitis, infestation level and parasite stage. Results: 40.3 percent (188) of the children presented Pediculosis; the six schools studied were affected. Pediculosis infestation in girls reached 55.2 percent (138/250) and 23 percent (50/217) in boys. Conclusion: The school population of Arica is significantly affected by Pediculosis and its prevalence is similar to other regions.

Variabilidad genética de poblaciones en cautiverio de Crocodylus moreletii (Crocodylia: Crocodylidae) mediante el uso de marcadores microsatelitales / Genetic variability in captive populations of Crocodylus moreletii (Crocodylia: Crocodylidae) using microsatellites markers

Genetic variability in captive populations of Crocodylus moreletii (Crocodylia: Crocodylidae) using microsatellites markers. Crocodylus moreletii, an extinction threatened species, represents an emblem for tropical ecosystems in Mexico. Surprisingly, there is a lack of information about their genetic constitution, which should be evaluated for a proper management ex situ and for making decisions on the release of crocodiles into natural habitats. The aim of this study was to characterize and compare the genetic variability of four populations of C. moreletii (two wild versus two born ex situ). Through PCR were amplified seven microsatellite polymorphic loci, however a heterozygote deficit, diminished by the presence of null alleles, was found in the populations (average H O=0.02). The AMOVA indicated that the highest proportion of genetic variability is within populations, and a limited genetic differentiation among populations (average F ST=0.03), probably due to high inbreeding index (average F IS=0.97). When comparing the genetic variability between and within other crocodilian species, we found that in C. moreletii is well below those reported. We concluded that the limited genetic variability in ex situ born populations is probably due to a founder effect derived from the social structure of their progenitors, and by the bottleneck effect, inferred by the limited effective population size, that historically characterizes their natural distribution in wild populations.

Crocodylus moreletii representa un emblema para los ecosistemas tropicales de México pero actualmente está amenazada por extinción. Sorprendentemente, hay una falta de información de su constitución genética, que debe ser evaluada para un manejo apropiado ex situ y para toma de decisiones en la liberación de cocodrilos a su hábitat natural. El objetivo del estudio fue caracterizar y comparar la variabilidad genética de cuatro grupos poblacionales de C. moreletii (dos silvestres y dos nacidas ex situ). Mediante PCR se amplificaron siete loci de microsatélites polimórficos, sin embargo se encontró déficit de heterocigotos en las poblaciones (promedio H O=0.02) mermado por la presencia de alelos nulos. El AMOVA indicó que la mayor proporción de variabilidad genética se encuentra dentro de las poblaciones y una limitada diferenciación genética entre poblaciones (promedio F ST =0.03), probablemente debida al alto índice de endogamia (promedio F IS=0.97). Al comparar la variabilidad genética inter e intra especies de cocodrilianos, encontramos que en C. moreletii está muy por debajo de los reportados. Se concluye que la limitada variabilidad genética de las poblaciones nacidas ex situ probablemente se debe al efecto fundador derivado de la estructura social de sus progenitores, y de las poblaciones silvestres, por el efecto cuello de botella, inferido por el limitado tamaño efectivo de población que presentó históricamente en su distribución natural.

Vegetative compatibility and heterokaryon formation between different isolates of Colletotrichum lindemuthianum by using the nit mutant system

Colletotrichum lindemuthianum, the causative agent of bean anthracnose, is one of the most common pathogens leading to expressive damage to plants beyond presenting noticeable variability. The knowledge on vegetative compatibility groups (VCGs) is of particular interest in asexual fungi as they subdivide the population in groups that can exchange genetic information via heterokaryosis and the parasexual cycle. Among the techniques used in studies about vegetative compatibility groups, the obtainment of nit mutants is apparent. This paper is aimed at obtaining heterokaryons between different isolates of C. lindemuthianum, grouping them in VCGs and evaluating their genetic variability by using the nit mutants system. Nit mutants were obtained from 20 single spore isolates. The mutants were phenotypically classified and paired for complementation and formation of heterokaryons so as to group them in VCGs. Seventeen mutants from the different phenotypic-rates were recovered: nit1, nit2, nit3 and nitM. At the same time, 10 mutants were selected for pairing and division of the anastomosis groups. Nine heterokaryons were obtained and the isolates were divided into 9 vegetative compatibility groups. In the combinations for the formation of anastomosis, 31 compatible combinations and 24 incompatible combinations were observed. It was concluded that the methodology used to select nit mutants in C. lindemuthianum made it possible to determine the vegetative compatibility groups and that such a technique was adequate to prove genetic variability.

Effects of low-dose lipopolysaccharide on cell apoptosis, proliferation, and insulin secretion in NIT-1 β-cells / 中华内分泌代谢杂志

ObjectiveTo investigate the effects of lipopolysaccharide ( LPS ) on cell apoptosis,proliferation, and insulin secretion in a β-cell line, NIT-1. MethodsNIT-1 cells were stimulated with 1 μg/ml LPS for 0-120 h. Cell apoptosis was evaluated by Hochest33342 staining and Annexin V/PI flow cytometry. Cell proliferation was evaluated by CCK-8 and BrdU assay. Intracellular insulin content, basal insulin secretion, and glucose-stimulated insulin secretion(GSIS) were detected by RIA. The IRS-2 tyrosine phosphorylation was determined by Western blot. ResultsCell apoptosis was not significantly changed by treatment with LPS for 120 h. Cell proliferation was stimulated by LPS before 48 h, and inhibited after 96 h. Intracellular insulin content or GSIS was not altered, but basal insulin secretion was decreased significantly by LPS after 48 h ( all P＜0.01 ). LPS decreased the tyrosine phosphorylation level of IRS-2 ( 0. 45 ± 0. 08 vs 0. 22 ± 0. 06, P ＜ 0. 05 ) and stimulated IκBα phosphorylation. Pretreatment with a specific IκBα phosphorylation inhibitor, Bay1 1-7082 for 1 h, remarkably blunted the LPS-induced phosphorylation of IκBα and cell proliferation( both P＜0.01 ). ConclusionsLow-dosages of LPS regulate proliferation and basal insulin secretion of NIT-1 β-cells, in which activation of NF-κB and inhibition of IRS2 tyrosine-phosphorylation may be involved.

Abnormalities in glucose metabolism and pancreatic endocrine function induced by 5-fluorouracil in vitro and in vivo / 肿瘤

Objective: To explore the effects of 5-fluorouracil (5-FU) on glucose metabolism and the endocrine function of pancreatic beta-cells. Methods: The NIT-1 cells were treated with different concentrations (5.0, 10.0, 20.0 and 40.0 mg/L) of 5-FU. The insulin concentration in the culture medium was determined by radioimmunoassay. Twenty Wistar rats were divided into 5-FU group (intraperitoneally administered with 5-FU at a daily dose of 20 mg/kg for 5 days) and the control group (intraperitoneally administered with normal saline). Glucose tolerance test was performed on d 2 and d 7 following 5-FU administration, and the plasma insulin concentration was determined by radioimmunoassay. Results: The high glucose (16.7 mmol/L)-induced release of insulin from NIT-1 cells was inhibited by 5.0 to 20.0 mg/L of 5-FU in a dose-dependent manner (P0.05). Postprandial plasma insulin level at 60 min following a glucose load on d 7 was significantly lower than that of the control group on d 2 (25.32 ± 6.07 vs 33.98 ± 4.94, P<0.05). The peak secretion time of plasma insulin was delayed to 120 min. Conclusion: Hyperglycemia following 5-FU treatment appears to be related to a relative deficiency in insulin secretion response to glucose stimulation. Copyright© 2011 by the Editorial Board of Tumor.

Use of oval conidia as a tool to assess the genetic transfer among Colletotrichum sublineolum mutants

The aim of this study was to assess oval conidia production in mutants for the reductase nitrate gene in C. sublineolum and verify the possibility of using them as a tool to transfer the genetic material in this species. The mutants used in the present study lost the ability to form falcate conidia but all produced oval conidia. The number of nuclei by conidium was evaluated. Oval conidia were efficient in the heterokaryon formation and these heterokaryons in liquid culture medium they also produced oval conidia in abundance. Recombinants were obtained and the genetic exchanges were confirmed by the RAPD analyses.

O fungo Colletotrichum sublineolum agente causal da antracnose em sorgo mostra dimorfismo conidial, produzindo conídios falcados ou ovais dependendo das condições de cultivo. Conídios falcados têm sido usados para obter heterocários entre mutantes complementares. Muitas linhagens de C. sublineolum não produzem conídios falcados ou produzem em quantidades insuficientes para analises. O objetivo do presente estudo foi avaliar a produção de conídios ovais em mutantes para o gene da nitrato redutase e verificar a possibilidade de sua utilização como ferramentas na transferência de material genético nesta espécie. Os mutantes usados neste estudo perderam a capacidade de produzir conídios falcados, porém todos produziram conídios ovais. O número de núcleos por conídio foi determinado. Conídios ovais foram eficientes na formação de heterocários e os heterocários quando cultivados em meio líquido também produziram conídios ovais em abundância. Recombinantes foram obtidos e as trocas genéticas foram confirmadas pelas análises de RAPD.

Effect of 11β-hydroxysteroid dehydrogenase type 1 gene silencing on glucose stimulated insulin secretion of pancreatic β cell line NIT-1 / 基础医学与临床

Objective To investigate the effect of small interference RNA (siRNA) targeting at 11β-hydroxysteroid dehydrogenase type 1 on the glucose-stimulated insulin secretion (GSIS) in pancreatic β cell line NIT-1 cell.Methods siRNA plasmid vectors specifically targeting at 11β-HSD1 gene were constructed,named as olig886,oligo866 and scrabble control for oligo886,then tansfected into NIT-1 cells.The expression of 11β-HSD1 was detected by RT-PCR and Western blot.O1igo886 vector was transfected into the NIT-1 cells in 25 mmol/L glucose concentrations medium.The insulin secretion level was measured in GSIS test.Results After treatment with 11β-HSD1 siRNA,the mRNA level of 11β-HSD1 in NIT-1 cell was decreased by 78.1%±2.9% and 51.7% ±2.7% inolig886 and oligo866 group respectively.The protein of 11β-HSD1 were decreased by 82.2% ±2.1% and 56.5%±2.0 % respectively.After transfected by olig 8 8 6 vector,the insulin secretion increased in NIT -1 cell.Conclusion 11β-HSD1 gene silencing may improve GSIS in NIT-1 cell 11β-HSD1 regulate local glucocorticoid metabolism in pan-creatic islet and affect the function of insulin secretion.

Effect of globular adiponectin on the function of NIT-I cells under high glucose medium / 中华内分泌代谢杂志

The effect of globular adiponectiin (gAd)on the function of NIT-1 cells under high glucose medium was investigated. The results showed that gad could completely block the increase of NADPH oxidase components p47phox expression and recover mRNA expression of pancreatic duodenal homeobox-I ,paired box gene 6,glucose transpoter 2,and glucokinase except neurogenic differentiation factor 1 (P<0.05 or P<0.01). Whereas,impaired insulin secretion and mRNA expression at high glucose concentration were not significantly improved by gAd.

Effect of survivin gene-transfection on cytokine-induced apoptosis in NIT-1 cells / 中华内分泌代谢杂志

Overexpression of survivin may partly protect the NIT-1 cells(mouse insulin-secreting cells) from cytokine-induced apoptosis.In addition, NIT-1 cells transfected with survivin had an slightly improved response of insulin secretion to glucose stimulation.

Cierre de conducto arterioso persistente con dispositivo Nit-Occlud: Experiencia de 13 casos / Patent ductus arteriosus closure with a Nit-Occluded device: Experience with 13 cases

Se presenta la experiencia de 13 pacientes con diagnóstico de persistencia de conducto arterioso permeable tratados por vía percutánea con un dispositivo de nitinol (aleación de níquel y titanio), llamado Nit-Occlud. Fueron 10 mujeres y 3 hombres con edad promedio de 23.2 ±21.1 años. La angiografía en la aorta descendente previa al cierre reportó diámetro promedio del conducto arterioso de 3.8 ± 0.8 mm y la morfología del mismo fue de 10 conductos tipo A, 2 tipo E y 1 tipo C según la clasificación de Krichenko. El tamaño del oclusor seleccionado se basó en el diámetro de la ámpula aórtica y del sitio más estrecho del conducto. Se utilizaron 8 dispositivos 11x6 mm, 4 dispositivos 9x6 mm y 1 dispositivo 7x6 mm. Tres pacientes (25%) mostraron oclusión del 100% en forma inmediata (15 min después de la liberación del dispositivo), 7 (53%) presentaron fuga leve y difusa y sólo 2 pacientes (16.6%) mostraron fuga moderada a severa. Un paciente presentó migración del dispositivo a la rama izquierda de la arteria pulmonar (RIAP), se logró su rescate por vía percutánea enviándose el paciente posteriormente a cirugía. En el seguimiento ecocardiográfico a las 24 h 9 pacientes (69%) mostraron oclusión del 100%. A los 6 meses de seguimiento 10 tienen oclusión al 100% y sólo 2 pacientes tienen fuga residual leve (15%). Un paciente desarrolló cuadro de endocarditis infecciosa que finalmente provocó su deceso. Se concluye que el dispositivo Nit-Occlud es útil para el cierre de esta patología aunque hace falta mayor experiencia.

We report our experience with the percutaneous closure of patent ductus arteriosus with a Nit-Occlud device made of Nitinol (Nickel-Titanium alloy) in 13 patients; 10 women and 3 men, average age of 23.2 ±21.1 years. Average diameters were 3.8 + 0.8 mm and the morphologies, according to Krichenko's classification, were 10 type A, 2 type E, and 1 type C. The device was selected according to the aortic ampule and the narrowest part of the ductus. We implanted 8 occluders of 11 x 6 mm, 4 of 9 x 6 mm, and 1 of 7 x 6 mm. In three patients (25%) total occlusion was observed 15 minutes after implantation, in seven (54%) a trivial leak was observed, and in only two patients (16.6%) was the leak moderate to severe. In on patient, the occluder migrate to the pulmonary artery trunk and was successfully removed percutaneously; the patient was subjected then to surgical closure. Echocardiography follow-up 24 h later showed total occlusion in nine patients (69%). Six months after the procedure, two patient presented trivial leak (15%). One patient developed an endocarditic infection and died. We concluded that this occluder might be useful for this pathology, but further studies must be done.

Mechanisms of PGF_ (2?) on the glucose-induced insulin secretion in NIT-1? cells / 中国病理生理杂志

0.05). However, in the presence of 16.5 mmol/L glucose, PGF_ 2? increased significantly in insulin secretion (P

SELECTION AND IDENTIFICATION OF NITRATE NON-UTILIZING MUTANTS OF COLLECTOTRICHUM GLOEOSPORIOIDES / 微生物学通报

Eight isolates of Colletotrichum gloeosporioides isolated from host plants Cunninghamia lanceolata lanceolata and Euonymus japonichum, respectively, were cultured on MMC medium containing KCIO3 to select chlorate-resistant and nitrate non-utilizing mutants (Nit). All the Nit mutants obtained by this way belong to one of 3 kinds of the following phenotypes: the nitrate reductase structural locus (nit1), the nitrate-assimilation pathway-specific regulatory locus(nit3), and the molybdenum-containing cofactor locus(nitM). The higher mutation frequency on MMC amended with increasing concentration of KCIO3 was induced, and various nitrogen sources were able to influence production of the phenotypic classes. Seven of the 8 isolates tested were self-compatibility in which the two mutants with different phenotypes from the same isolate could genetically complement. The two isolates frail C. lanceolata belong to the same vegetative compatibility group (VCG), the other six isolates belong to distinctive VCGs.

Effects of PGF2? on the glucose-stimulated insulin secretion in NIT-1 beta cells / 中国药理学通报

Aim To investigate the effects of PGF_(2?) upon glucose-stimulated insulin secretion and the calcium response in NIT-1 beta cells.Methods Using the radioimmunoassay(RIA),the amount of PGF_(2?) augmentation of glucose-stimulated insulin secretion was determined in different conditions and the confocal laser scanning methods by Fluo-3AM as a fluorescent probe were used to analyze the NIT-1 beta cell intracellular calcium response in correlated various terms.Results In the presence of 16.5 mmol?L~(-1) glucose,PGF_(2?)(0.1,1,5 ?mol?L~(-1)) dose-dependently augmented glucose-induced insulin secretion in NIT-1 beta cells,especially at 5 ?mol?L~(-1)(P0.05).Meanwhile,Exposure of the NIT-1 cells to 5 ?mol?L~(-1) PGF_(2?) induced a rapid increase of intracellular calcium(P