Evaluation of immunocytochemistry on destained giemsa stained smears as an alternative to conventional technique

Introduction: Protocol for immunocytochemical (ICC) staining in May-Grünwald Giemsa (MGG)–stained smears has been difficult to establish. It is the need of the hour to be able to use prestained slides for ICC in specific cases to deliver timely diagnoses and reduce inconvenience to patients. Aims and Objectives: To evaluate and compare the use of MGG-stained smears for the purpose of ICC, after de-staining and saline rehydration to that of routine standard ICC. Materials and Methods: A prospective study was conducted on 40 FNAC samples: 25 cases of breast disease and 15 cases of reactive lymphoid hyperplasia known to express pancytokeratin and leukocyte common antigen (LCA)/CD45, respectively. Air-dried smears of each case were stained by standard MGG stain and after the report was dispatched, one smear was selected and sent for ICC. The smears were analyzed to determine the overall result and grade each smear semi-quantitatively with respect to staining-intensity, stain-localization, staining-uniformity, counter-staining, and background-staining. Observations and Results: The proposed protocol was inferior to conventional ICC in all the parameters, more pronounced in pancytokeratin than LCA/CD45. Only 8% of air-dried smears stained for pancytokeratin showed optimal stain intensity (as opposed to 44% of wet-fixed smears), whereas only 14.3% of air-dried smears were optimally stained for LCA (as opposed to 85.7% of wet-fixed smears). Conclusion: The proposed protocol of de-stained Giemsa smears as an alternative to conventional technique for ICC was unsuccessful in giving satisfactory results.

Cromosomopatías en pacientes con cáncer sometidos a radioterapia con cobalto 60

Objetivo: Describir las aberraciones citogenéticas que pueden ser observadas por medio de la técnica Giemsa en fluorescencia y encontradas en pacientes con cáncer antes y después de ser sometidos a tratamiento con radioterapia. Métodos: Se analizó un mínimo de 200 metafases en primera división mitótica antes y después del tratamiento de radioterapia en nueve pacientes que asistieron a la sección de radioterapia del Hospital San Juan de Dios Costa Rica. En cada caso se contabilizó cada tipo de cromosomopatía por medio de la prueba de Giemsa en fluorescencia y utilizando bromodeoxiuridina y naranja de acridina. Resultados: Las cromosomopatías producidas por radioterapia se observaron tanto antes como después del tratamiento sin embargo destacó el incremento en la frecuencia de los cromosomas dicéntricos y anillos céntricos una vez finalizada la terapia. La frecuencia de fracturas cromatídicas de asociaciones satelíticas y de alteraciones morfológicas no se ve afectada por la radioterapia. Uno de los participantes presentó un recuento mitótico bajo. Conclusión: La radioterapia aumenta significativamente la frecuencia de los cromosomas dicéntricos y dicéntricos más anillos en la muestra en estudio. Este trabajo es relevante por ser el primer estudio en Costa Rica en el que se analizan los cromosomas dicéntricos como biomarcadores de exposición a radiaciones ionizantes mediante la prueba de Giemsa en fluorescencia y utilizando bromodeoxiuridina y naranja de acridina.

Aim: The objective of this study was to describe the before and after cytogenetic aberrations found in current patients of radiotherapy. This can be observed through the technique called "Giemsa in fluorescence" Methods: A minimum of 200 metaphases were analyzed in the first mitotic division in 9 patients. The patients where observed before and after radiotherapy treatment at the San Juan de Dios Hospital in Costa Rica. In each case any type of chromosomopathy was counted using the "Giemsa in fluorescence" test as well as Bromodeoxyuridine and acridine orange. Results: The chromosomopathies are observed before and after treatment with radiotherapy. The treatment seems to change the frecuency increasing the dicentric chromosomes and centric rings after the treatment. The frequency of chromatid fractures satellite associations and morphological alterations were not affected by radiotherapy. Conclusion: The chromosomopathies produced by radiotherapy were observed both before and after treatment with variations in their frequency. After radiotherapy dicentric chromosomes and dicentric chromosomes plus rings frequencies increased significantly. A low mitotic count was present this could have been the result of radiation on the bone marrow or by the cell repair and apoptosis system. The standardized " Fluorescence Plus Giemsa" test using Bromodesoxyuridine and acridine orange was used for the fiesta time in Costa Reica. This allowed for the measurement of radiation exposure used in the treatment or detection of diseases and cancer in pacients.

Combined rapid urease test and histology for the diagnosis of helicobacter pylori infection

Significance@#Accurate detection of Helicobacter pylori (HP) is essential for the diagnosis of HP infection. The use of antibiotics and proton pump inhibitors (PPI) may result in false-negative rapid urease test (RUT) results. We aimed to determine the sensitivity and specificity of RUT compared with histology and assess the detection rate of combined RUT and histology for HP infection. @*Methodology@#Retrospective data collection was performed on 192 patients who were tested for both RUT and histology at the time of upper endoscopy from 2017 to 2018. At least two gastric biopsies (1 from corpus, 1 from antrum) were taken each for RUT and histology. The endoscopy was performed by a single gastroenterologist and a single pathologist was responsible for interpreting the histology with hematoxylin and eosin (H&E) and Giemsa stain. The gold standard test for the diagnosis of HP infection was histology. Demographic profile, RUT and histology results were reviewed. Tests for diagnostic accuracy were computed using SPSSv23. @*Results@#192 patients were tested for RUT and histology. 52(27.1%) were males and 140(72.9%) were females with a mean age of 54±17 years. Epigastric pain was the most common indication (42.7%). 24(12.5%) patients tested positive for HP infection. Among these; 16(8.3%) tested positive for both RUT and histology(true-positive), while 8(4.2%) tested negative for RUT but had positive histology(false-negative). 6 out of 8(75%) patients with false negative results had PPI use. The sensitivity and specificity of RUT for the diagnosis of HP infection were 66.7 and 98.2%, respectively. While the positive and negative likelihood ratio were 37.3 and 0.34, respectively with a diagnostic odds ratio of 110. @*Conclusion@#The HP detection rate of RUT combined with histology increased by 33% compared with RUT alone. RUT is a highly specific test for diagnosing HP infection. Given its modest sensitivity, histology plays an important role in the diagnosis of HP infection, especially in patients taking PPIs. We recommend doing histology when RUT is negative to increase the HP detection rate.

A prospective study of proportion of Helicobacter pylori infection in patients presenting with dyspeptic symptoms in S. Nijalingappa Medical College and H.S.K. Hospital, Bagalkot

Background: More than half of the world’s population is infected with Helicobacter pylori, the primary cause of chronic gastritis. Chronic gastritis is associated with peptic ulcer and in advanced stages with an increased risk of developing gastric adenocarcinoma. The aim of the study was to determine the prevalence of Helicobacter pylori infection in dyspeptic patients and its relationship with gastroduodenal pathologies using gastric biopsy histology.Methods: A cross sectional prospective study was conducted from 1st January 2018 to 30th June 2019. It included out-patients and in-patients, presenting with dyspeptic symptoms and undergoing upper GI Endoscopy, at S. Nijalingappa Medical College and HSK hospital, Bagalkot, Karnataka, India. Endoscopic impressions were noted. In case of any abnormal findings on upper gastrointestinal endoscopy, endoscopic biopsy was obtained. Histopathological assessment of gastric mucosa was done after staining with H and E stain and Giemsa stain. The histopathological diagnosis of Helicobacter pylori infection in biopsy specimen will be mainly done using the Modified Giemsa stain.Results: 105 patients with dyspepsia were studied in total. Out of which, 44 patients (41.9%) were detected positive for Helicobacter pylori infection.Conclusions: The burden of Helicobacter pylori infection in patients with dyspepsia was high. Early diagnosis and eradication of Helicobacter pylori not only improves symptoms but also help to prevent complications associated with Helicobacter pylori infections.

Geospatial Modeled Analysis and Laboratory Based Technology for Determination of Malaria Risk and Burden in a Rural Community

Introduction:Geographical Information System (GIS) has proven to be very useful for large scale mapping of ecosystems, land use and cover, disease prevalence, risk mapping and forecasting. GIS establish relationship or link between vector borne diseases and associated environmental factors thereby providing explanation for spatial distribution pattern, possible causes of diseases outbreak andimplications on the community.Aims and Objectives:Our approach in this study was to define and identify areas and places that are exposed to Malaria risk through proximity analysis and to compare geospatial risk with laboratory diagnosed malaria epidemiology. Methodology:Garmin GPS was used to capture the geographic coordinates of six (6) selected settlements and overlaid with georeferenced and processed satellite images in the study area. GIS modeling was performed on risk factors using weighted overlay technique to produce malaria risk map. A total of One hundred and thirty-five (135) vulnerable individuals were diagnosed for Malaria with light Olympus microscope and rapid diagnostic kit (RDT). Data were entered and analyzed using R-Package for Statistical Computing and Graphics.Results:Proximity to malaria risk follows relatively the order Apodu > Central Malete > Elemere > KWASU Campus > Gbugudu. Apodu being the largest place with proximity to malaria risk, within 500m radius. The risk index increases as one move away from the center of the settlement. The possible explanation for this high risk could be the presence of pond / lake in Apodu. This is a good breeding site for mosquito couple with dense vegetation as one move away from the centre of the settlements. Unlike Apodu, Gbugudu was at medium risk at 100m buffer (60%) but the risk index decreases as one move away from the settlement centre. The absence of thick vegetation and presence of numerous open farms and partly cultivated farmlands on the eastern part could have been responsible for reduction in risk index. Dense vegetation and ponds were observed within Apodu, while Central Malete was built up with dense vegetation are possible reasons for the high-risk index, while settlements within 1 km radius around KWASU campus recorded lower risk index possibly dueto low vegetation. The geospatial malaria risk analysis correlates with the laboratory-based test results. RDT kits and light microscopy results showed Apodu having the highest malaria prevalence with 46% and 58.7% followed by Elemere 41% and 30.3% respectively. When calculating prevalence by aggregating results across all communities, Apodu still had the highest malaria prevalence for the whole region. RDT and light microscopy results combined for all communities had Apodu with malaria prevalence of 21.48% and 27.4% followed by Elemere with 11.85% and 12.5% respectively. Gbugudu had the least malaria prevalence within the region with 3.7% and 7.4% respectively.Discussion and Conclusion:Findings of this study showed dense vegetation and ponds within Apodu, Elemere and Central Malete served as good breeding site for mosquitoes and were responsible for the high-risk index at these areas. Settlements within 1 km radius around KWASU campus recorded lower index possibly due to low vegetation. Results from this study indicate that the degree of malaria parasitaemia in the three major settlements correlates directly with the remote sensing data

Perspectivas en investigación: células HEp-2 infectadas con EB's de Chlamydia trachomatis serovar 2 (VR-902B) / Chlamydia trachomatis: Research Perspectives. HEp-2 Cells Infected with EB's from Chlamydia trachomatis serovar 2 (VR-902B): Research Perspectives

Resumen Chlamydia trachomatis (C. trachomatis) es una bacteria Gram negativa inmóvil, caracterizada por ser un microorganismo intracelular obligado y por poseer un ciclo reproductivo en el que puede distinguirse una forma infecciosa extracelular metabólicamente inerte (cuerpo elemental - EB's), y una forma no infecciosa intracelular y activa (cuerpo reticulado - RB's). C trachomatis se caracteriza por causar infección en humanos, está relacionada con enfermedades de transmisión sexual e infecciones oculares; por lo que puede conllevar a secuelas de interés, si no se da un tratamiento oportuno. El objetivo de este estudio fue optimizar el modelo de infección de C. trachomatis en células HEp-2 con cuerpos elementales (EB's) de C. trachomatis serovar L2. Inicialmente, se establecieron las condiciones para el crecimiento adecuado de las células HEp-2 en tiempo y con una confluencia del 90%, para continuar con la optimización de un protocolo de infección. La infección fue confirmada a partir de la coloración con Giemsa permitiendo evaluar características morfológicas tanto de las células HEp-2 sin infectar e infectadas, y así mismo, de los cuerpos elementales de C. trachomatis. Finalmente, se corroboró la infección con la técnica de inmunofluorescencia directa que detecta la proteína de membrana MOMP de C. trachomatis. Tras los ensayos realizados se evidenció la presencia de cuerpos elementales próximos y dentro del citoplasma celular, así como células vacuoladas y daño celular causado por la infección.

Abstract Chlamydia trachomatis (C. Trachomatis) is a Gram negative unmoving bacterium, characterized by being an obligate intracellular microorganism and having a reproductive cycle in which a metabolically inactive extracellular infectious form (elementary body - EB's) can be distinguished from an intracellular active and non-infectious form (reticulated body - RB's). C trachomatis is characterized by causing infection in humans, is related to sexually transmitted diseases and eye infections, so it can lead to sequelae of interest if timely treatment is not given. The objective of this study was to optimize the infection model of C. trachomatis in HEp-2 cells with elementary bodies (EB's) of C. trachomatis serovar L2. Initially, the conditions for the adequate growth of HEp-2 cells were established in time and with a confluence of 90%, to continue with the optimization of an infection protocol. The infection was confirmed from the staining with Giemsa allowing to evaluate morphological characteristics of both uninfected and infected HEp-2 cells and also of the elementary bodies of C. trachomatis. Finally, the infection was corroborated with the direct immunofluorescence technique, that detects the C. trachomatis MOMP membrane protein. After the tests were performed, the presence of elementary bodies nearby and within the cellular cytoplasm was evidenced, as well as vacuolated cells and cellular damage caused by the infection.

Evaluación clínica y coprológica en sujetos sintomáticos y asintomáticos con infección por Blastocystis spp / Clinical and coprological assessment in symptomatic and asymptomatic subjects with infection with Blastocystis spp

Para evaluar los aspectos morfológicos, clínicos y de laboratorio, en sujetos sintomáticos y asintomáticos con infección por Blastocystis spp. se analizaron 250 muestras fecales de individuos del Hospital Universitario "Antonio Patricio de Alcalá", Cumaná, estado Sucre, previo consentimiento informado. Cada espécimen fecal fue analizado mediante examen directo, la cuantificación del cromista se hizo en diez campos consecutivos de 40X, el análisis morfométrico mediante frotis fecales teñidos con Giemsa observados a 100X, midiendo las estructuras con un micrómetro ocular. Del total de individuos evaluados 32,00% presentó infección por el cromista, 31,25% con monoinfección con más de 5 parásitos por campo de 400X, de los cuales 71,25% de ellos fueron sintomáticos. El morfotipo mayormente identificado fue el de cuerpo central, aunque en una sola muestra de consistencia líquida, se identificó también el de resistencia, hallazgo que no fue posible en el examen directo. En lo concerniente al análisis morfométrico, al evaluar los diámetros de los morfotipos de cuerpo central en pacientes con y sin síntomas, mediante la prueba no paramétrica W Mann-Whitney, se evidenció correspondencia entre ellos (W= 5360,0; p<0,05). El estudio morfológico, la cuantificación del número de parásitos por campo y la utilización de técnicas de tinción complementarias, pueden contribuir a optimizar el diagnóstico de laboratorio.

To evaluate morphological, clinical and laboratory aspects in symptomatic and asymptomatic subjects with Blastocystis spp. 250 faecal samples were analyzed from individuals of the University Hospital "Antonio Patricio de Alcalá", Cumaná, Sucre state, with prior informed consent. Each fecal specimen was analyzed by direct examination, Ritchie and Kinyoun method; The quantification of the chromist was done in ten consecutive 40X fields, the morphometric analysis by fecal smears stained with Giemsa observed at 100X, measuring the structures by an ocular micrometer. Of the total of individuals evaluated 32.00% presented infection by the chromist, 31.25% with monoinfection with more than 5 parasites per field of 400X, of which 71.25% of them were symptomatic. The most identified morphotype was that of the central body, although in a single of liquid consistency, resistance morphotype was also identified, a finding that was not possible in the direct examination. Regarding the morphometric analysis, when evaluating the diameters of the morphotypes of the central body in patients with and without symptoms, by means of the non-parametric W Mann-Whitney test, the correspondence between them was evidenced (W= 5360,0; p<0,05). The morphological study, the quantification of the number of parasites per field and the use of complementary staining techniques, may contribute to optimize the laboratory diagnosis.

Comparison of HE and Giemsa staining in diagnosis of bacterial and allergic conjunctivitis in children / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM:To explore the diagnostic effect of hematoxylin-eosin staining(HE)and Giemsa staining in the diagnosis of bacterial and allergic conjunctivitis in children. <p>METHODS:Totally 422 children with conjunctivitis diagnosed by conjunctivitis from the ophthalmology department of our hospital during 2016-10/2019-10 as the research objects. HE and Giemsa staining methods were used to stain the conjunctival scratches, and the staining results were used to diagnose bacterial/allergic conjunctivitis. Observe the positive detection rate of the two staining results for bacterial/allergic conjunctivitis and the staining situation. <p>RESULTS: The positive rate(33.0%)and coincidence rate(63.6%)of HE staining for the diagnosis of bacterial conjunctivitis were significantly lower than Giemsa staining(90.7% and 88.8%, <i>P</i><0.001), while the positive rate of allergic conjunctivitis was not significantly different 90.8% <i>vs </i>87.2%, <i>P</i>>0.05).<p>CONCLUSION: The Giemsa staining method can accurately diagnose bacterial conjunctivitis in children and the method is simple. Both HE and Giemsa staining methods have good diagnostic effects on allergic conjunctivitis, which can provide a basis for improving the clinical diagnosis efficiency and early treatment options.

Evolutionary analysis of genus Channa based on karyological and 16S rRNA sequence data

A wide range of diploid number of chromosomes and the body size of Channa congeners are useful combination of characters for studying the factors controlling the body size. In this study, the karyological information was superimposed on the evolutionary tree generated by 16S rRNA mitochondrial gene sequences. Here, the metaphase chromosome complements stained with Giemsa, AgNO3 and CMA3 were prepared from six snakehead murrel fish species collected from northeast India. The diploid chromosome numbers and the fundamental arms of C. aurantimaculata (2n = 52, NF = 98), C. gachua (2n = 56, NF = 84), C. marulius (2n = 44, NF = 58), C. orientalis (2n = 52, NF = 74), C. punctata (2n = 32, NF = 60) and C. striata (2n = 40, NF = 48) were calculated by the analysis of metaphase chromosome complements. Both methods of nucleolar organizer region (NOR) localization, silver nitrate and chromomycin A3, revealed NOR pairs of 1, 2, 3, 1, 4 and 3 in C. aurantimaculata, C. gachua, C. marulius, C. orientalis, C. punctata and C. striata, respectively. The subject species showed primitive type of asymmetrical chromosomes, except the C. punctata. The variation in 2n for C. orientalis (2n = 52, 78) and C. gachua (2n = 52, 78, 104) of a complete haploid set indicates the possibility of either ploidy change in . orientalisC and C. gachua, if we consider 2n = 52 or the Robertsonian rearrangements in different populations of these two species. The chromosome evolution tree was constructed on 16S rRNA ML-phylogenetic tree using ChromEvol 1.3. The analysis of chromosome evolution explained the loss or gain of chromosome, duplications or semiduplications mechanism. For time scaling the chromosomeevolution, the node age of available 16S rRNA gene of Channa species were estimated, which was also used for estimating the time when chromosomal changes occurred in context of geological time-scale.

Ultrasound-guided fine-needle aspiration cytology of gallbladder masses – A solution to diagnostic dilemmas

Background: Advances in medical technology has revolutionized patient care. Ultrasound (USG)-guided fine-needle aspiration cytology (FNACs) is enlarging the realm of diagnostic cytology. This merger has increased the diagnostic accuracy especially in inaccessible body sites like the gall bladder (GB) masses. Impalpable, deep, and small lesions are now no longer a dilemma, thanks to image-guided FNACs. Objectives: The objective of the study was to study the ultrasound-guided FNACs of GB masses for a 10 year period and evaluate the efficacy of ultrasound-guided FNACs in GB lesions. Materials and Methods: USG-guided FNACs were collected and analyzed over a 10 year period (2007–2016) in the Department of Pathology North Eastern Indira Gandhi Regional Institute of Health and Medical Sciences (NEIGRIHMS), Shillong. This is a retrospective study and is in accordance with the ethical standards as laid by the Helsinki Declaration, 2000. Descriptive statistics were used for analysis. Results: The age ranges from 27 to 83 years and the mean age was 53.2 years. The majority were females which constituted around 94.3% (50) and males constituted around 5.7% (3). Out of all the adequate cases (42 in number) that were evaluated, 37 (88%) were malignant and 5 (12%) were benign and inflammatory; 11 out of 53 cases (20.8%) were inadequate. Conclusion: Although FNAC is established as a reliable diagnostic method, our study of 10 years has further proven that this test has stood the test of time, and with our high diagnostic yield and in the hands of a dedicated radiologist and cytopathologist, it will prove to surpass its expectations.

Comparison of different stains in exfoliated oral mucosal cell micronucleus of potentially malignant disorders of oral cavity

Background: Micronucleus (MN) has been proved to be an important biomarker of genomic damage. Leishman Giemsa (LG) cocktail, being a relatively new staining technique, has not been used in exfoliative cytology. The aim of this study is to observe and compare the micronuclei (MN) frequency in potentially malignant disorders (PMDs) and also to compare the staining efficacy of May-Grünwald Giemsa (MGG), LG cocktail, and Papanicolaou (PAP) for micronuclei in exfoliated oral mucosal cells. Materials and Methods: Three smears were prepared from each 30 controls (buccal mucosa) and 120 patients (40 oral submucous fibrosis, 40 lichen planus, and 40 leukoplakia) clinically diagnosed with having one of the PMDs of the oral cavity stained with PAP, MGG, and LG cocktail stains. MN frequency (No. of MN/1000 cells) was evaluated and compared between the cases and the controls. Comparison between the three different stained smears was also made to determine the clarity and efficacy of the stains. Results: LG cocktail gave comparatively better results followed by PAP and MGG. Statistically significant results (P < 0.05) were obtained, using Mann–Whitney test for comparison of MN frequency between cases and controls. Conclusion: LG cocktail is an easy, cost- effective, and one step technique comparable to PAP staining; however, it warrants further study in its potential application in screening of oral cancer.

Comparative study of metachromatic staining methods in assessing the exfoliative cell types during oestrous cycle in Sprague-Dawley laboratory rats / Estudio comparativo de los métodos de tinción metacromática en la evaluación de los tipos de células exfoliativas durante el ciclo estral en ratas Sprague-Dawley de laboratorio

This study was aimed at comparing the commonly used metachromatic stains viz., Papanicolaou stain, WrightGiemsa, Toluidine blue and Methylene blue in the assessment of cell types of the oestrous cycle in rats. Eight female Sprague-Dawley rats aged 8-9 weeks were used for this assessment. Cotton Swabs were gently inserted in the animals vagina to obtain cells from which they were then transferred to glass slides for staining and evaluation under microscopy. The different cell types were compared for their morphological features and clarity of cellular detail under all four stains. The application, advantages and limitations of all stains were then discussed. It was concluded that the selection of the most effective stain in the assessment of vaginal cytology depends on their application to clinical or research which was based on the cellular detail of interest, time, cost and availability of each staining procedure.

El presente estudio tuvo como objetivo comparar las tinciones metacromáticas comúnmente utilizadas, Wright's-Giemsa, azul de toluidina, azul de metileno y tinción de Papanicolaou, en la evaluación de los tipos de células del ciclo estral en ratas. El estudio se realizó en ocho ratas hembras SpragueDawley, con edades entre 8 y 9 semanas, y se usaron hisopos vaginales de algodón para preparar portaobjetos. Los diferentes tipos de células se compararon por sus características morfológicas y claridad en las cuatro tinciones. La aplicación, ventajas y limitaciones de todas las tinciones fueron discutidas. Se concluye que la selección de la tinción más efectiva en la evaluación de la citología vaginal depende de su uso, es decir, clínico o de investigación, el detalle celular de interés, tiempo, costo y disponibilidad.

Morphoquantitative study of Rattus norvegicus submucosal plexus by different neuronal evidentiation histochemical techniques / Estudio morfocuantitativo del plexo submucoso de Rattus norvegicus por medio de diferentes técnicas histoquímicas para verificar características neuronales

Enteric nervous plexuses have been the object of several studies, specially the myenteric plexus whose studies describe its organization, functions and alterations. On the other hand, the submucosal plexus has been less studied and still needs descriptive studies. To analyze morphologically and quantitatively submucosal neurons of the jejunum of 90-day-old healthy rats using different techniques for neuronal staining as a way to provide normality data to compare with future experimental studies. Whole mount preparations of the jejunum were submitted to Giemsa, NADH-diaphorase and NADPH-diaphorase techniques to stain the total neuronal population, more metabolically active subpopulation and subpopulation of nitrergic neurons, respectively. Neurons of the submucosal plexus of adult rats are mainly organized in ganglia with varied sized and shapes. Giemsa technique stained 243.93 ± 7.68 neurons per mm2. Regarding the total population stained by Giemsa, NADH- diaphorase positive (139.09 ± 11.14/mm2) neurons represented 57 % and NADPH-diaphorase positive (18.17 ± 0.28/mm2) represented 7.5 %. The area of the cell body was bigger in nitrergic neurons (412.29 ± 150.22) than in the ones stained by Giemsa (254.71 ± 63.32) and NADH-diaphorase positive (243.98 ± 123.82).

El plexo nervioso entérico ha sido objeto de varios estudios, especialmente el plexo mientérico, cuyos estudios consisten en describir su organización, funciones y alteraciones. Por otro lado, el plexo submucoso ha sido menos investigado y todavía necesita estudios descriptivos. Para analizar morfológica y cuantitativamente las neuronas de la submucosa del yeyuno de ratas de 90 días de edad, se realizaron diferentes técnicas de tinción neuronales, en animales sanos, como una forma de proporcionar datos de normalidad y compararlo con futuros estudios experimentales. Se realizaron montajes con preparados enteros del yeyuno que fueron sometidos a las técnicas de Giemsa, de NADPH-diaforasa y NADH-diaforasa para teñir la población total neuronal, subpoblación más activa metabólicamente y subpoblación de neuronas nitrérgicas, respectivamente. Las neuronas del plexo submucoso de ratas adultas se organizan principalmente en los ganglios con variaciones de tamaño y formas. Con la técnica de Giemsa se tiñeron 243.93±7.68 neuronas por mm2. Con respecto a la población total teñida con Giemsa, fueron positivas para NADH- diaforasa en 139.09 ±11.14 / mm2 neuronas, representando el 57% y fueron positivas para NADPH-diaforasa en 18,17 ± 0,28 / mm2 neuronas, lo que representó el 7,5%. El área del cuerpo celular fue mayor en neuronas nitrérgicas (412,29 ± 150.22) que en las teñidas con Giemsa (254,71 ± 63,32) y NADH-diaforasa positivas (243,98 ± 123,82).

Trichomonas vaginalis infection in a low-risk women attended in Obstetrics and Gynaecology Clinic, Universiti Kebangsaan Malaysia Medical Centre

Objective: To investigate the presence of trichomoniasis among women attending the Obstetrics and Gynaecology Clinic, Universiti Kebangsaan Malaysia Medical Centre. Methods: A total of 139 high vaginal swabs were taken from the subjects and sent to the laboratory in Amies gel transport media. The specimens were examined for the presence of Trichomonas vaginalis using wet mount, Giemsa staining and cultured in Diamond's medium. Sociodemographic characteristics and gynaecological complaints were obtained in private using structured questionnaire applied by one investigator. Results: The median age was 32 years, with an interquartile interval of 9.96. Most of the subjects were Malays (76.9%) and the remaining were Chinese (15.1%), Indians (2.2%) and other ethnic groups (5.8%). One hundred and thirty eight (99.3%) of the women were married and 98.6%had less than 6 children. More than half (75.5%) of the women's last child birth was less than 6 years ago. Forty seven percent of them were involved in supporting administrative work and 64.7% of the women gave a history of previous or current vaginal discharge. Conclusions: The present study reported zero incidence rate of trichomoniasis. The low incidence rate was postulated due to all women who participated in this study were categorized into a low-risk group.

Protective effects of LiCl on synaptic atrophy induced by okadaic acid in differentiated SK-N-SH neuroblastoma cells / 中国药理学通报

Aim To explore the protective effects of lithium chloride ( LiCl ) on neurous injuries and phos-phorylation of tau protein at serine262 induced by okada-ic acid( OA) . Methods The neuroblastoma SK-N-SH cells were differentiated by all-trans-retinoic acid ( AT-RA) . The differentiated SK-N-SH cells were treated with OA to establish the Alzheimer′s disease cellular model. SK-N-SH cells′ viability and proliferation were measured by SRB test. Giemsa staining was used to observe cell morphology. The neurite length of SK-N-SH cells was measured by Image-Proplus software. Syn-aptophysin and phosphorylated tau protein at serine262 expression levels were tested by Western blot. Results The SK-N-SH cells which were treated with 10 μmol ·L-1 ATRA for 7 days displayed mature neuronal fea-tures. The synaptic length of SK-N-SH cells became longer. And the levels of serine262 phospho-tau was sig-nificantly elevated. 20~100 nmol·L-1 OA effectively inhibited the viability of differentiated SK-N-SH cells in a concentration-dependent manner and in a time-de-pendent manner. The OA treatment induced obvious synaptic atrophy in differentiated SK-N-SH cells. And the phosphorylation level of tau protein serine262 also greatly increased. The pretreatment with 10 mmol · L-1 LiCl significantly ameliorated the synaptic atrophy, the decrease of synaptophysin expression and the in-crease of tau phosphorylation at serine262 induced by OA in differentiated SK-N-SH cells. Conclusion LiCl could effectively inhibit OA-induced synaptic atro-phy in differentiated SK-N-SH cells, and it could also result in the increase of synaptophysin expression and the decrease of the phosphorylation of tau protein at serine262 .

Trichomonas vaginalis infection in a low-risk women attended in Obstetrics and Gynaecology Clinic, Universiti Kebangsaan Malaysia Medical Centre

Objective To investigate the presence of trichomoniasis among women attending the Obstetrics and Gynaecology Clinic, Universiti Kebangsaan Malaysia Medical Centre. Methods A total of 139 high vaginal swabs were taken from the subjects and sent to the laboratory in Amies gel transport media. The specimens were examined for the presence of Trichomonas vaginalis using wet mount, Giemsa staining and cultured in Diamond's medium. Sociodemographic characteristics and gynaecological complaints were obtained in private using structured questionnaire applied by one investigator. Results The median age was 32 years, with an interquartile interval of 9.96. Most of the subjects were Malays (76.9%) and the remaining were Chinese (15.1%), Indians (2.2%) and other ethnic groups (5.8%). One hundred and thirty eight (99.3%) of the women were married and 98.6% had less than 6 children. More than half (75.5%) of the women's last child birth was less than 6 years ago. Forty seven percent of them were involved in supporting administrative work and 64.7% of the women gave a history of previous or current vaginal discharge. Conclusions The present study reported zero incidence rate of trichomoniasis. The low incidence rate was postulated due to all women who participated in this study were categorized into a low-risk group.

Efficacy of FNAC in early diagnosis of prostatic carcinoma

Background: Prostatic carcinoma is one of the most important causes of mortality in elderly men mainly because of the late detection despite of the fact that it is a potentially curable disease. Fine needle aspiration cytology (FNAC) is an easy to perform outpatient procedure requiring no expensive equipment or anesthesia. Objectives: The present study was carried out in an attempt to evaluate the fine needle aspiration cytology in the diagnosis of carcinoma prostate. Materials and methods: The present study was performed on 27 patients admitted in the surgical wards, with complaints suggestive of prostatic disease, in whom there was found to be a suspicion of malignancy of the prostate gland. Results: Among 27 patients, 14 patients were diagnosed as prostatic malignancy on per rectal FNAC whereas, 19 patients were confirmed with prostatic cancer histologically. Out of 19 histologically confirmed cancer cases, 16 were also positive on FNAC i.e. 84.21% accuracy of FNAC in detecting prostatic malignancy. Conclusion: Fine needle aspiration cytology is easily available and inexpensive procedure. It is a reliable method in the diagnosis of prostatic cancer. Its positive results are relatively more reliable than the negative ones. It is an effective method in follow up of the cancer cases.

Histological identification of H. pylori stained by hematoxylin-eosin and Giemsa: review for quality control / Identificação histológica do H. pylori corado por hematoxilina-eosina e Giemsa: revisão para controle de qualidade

Introduction: Several special staining methods are available for H. pylori (Hp) identification in histological sections of chronic gastritis (CG), including the routine hematoxylin-eosin (HE) method. Some reports suggest that ancillary stains are not always needed to establish the diagnosis of Hp infection. In addition, the benefit of using them, when biopsies show minimal inflammation, is not clear. Objective: We performed a retrospective study to compare the usefulness of HE with Giemsa method for the histopathological diagnosis of Hp in tissue sections. Methods: Histological sections from 390 consecutive patients were reviewed. The patients were registered in the histopathology laboratory of Instituto Alfa de Gastroenterologia, Hospital das Clínicas da Universidade Federal de Minas Gerais (UFMG), Belo Horizonte, Brasil. They were divided in 4 groups according to the gastric inflammatory changes as follows: Group I, gastric mucosa with normal morphology or minimal inflammatory changes (n = 146); Group II, chronic gastritis (CG) with mild inflammatory activity (n = 101); Group III, CG with patent inflammatory activity (n = 123); Group IV, patients with atrophic body gastritis (n = 20). All histological sections were carefully evaluated by 2 examiners at the oil immersion objective (1000×). Results: The identification of Hp was positive by Giemsa and HE, respectively at: Group III, 111 (90.2%) and 93 (75.6%) patients (p < 0.01); Group II, 43 (42.6%) and 29 (28.7%) patients (p < 0.05). Hp was negative in Groups I and IV. Conclusion: The results show that Giemsa stain is superior to HE for histological identification of Hp in CG. Although Hp could be identified by HE stain in the majority of CG cases, a significant number of infected patients may be neglected, regardless the intensity of the inflammatory response. .

Introdução: Diversos métodos de coloração especial estão disponíveis para a identificação da bactéria H. pylori (Hp) em cortes histológicos. Entretanto, questiona-se a utilidade desses métodos na rotina diária dos laboratórios de patologia diagnóstica. Objetivos: Comparar a utilidade da coloração por hematoxilina-eosina (HE) com a do Giemsa para diagnóstico histopatológico do Hp. Materiais e métodos: Foram revistos os cortes histológicos de 390 pacientes consecutivos cadastrados no Laboratório de Histopatologia do Instituto Alfa de Gastroenterologia, Hospital das Clínicas da Universidade Federal de Minas Gerais (UFMG), Belo Horizonte, Brasil. Os pacientes foram divididos em quatro grupos: Grupo I - mucosa gástrica apresentando morfologia normal (n = 146); Grupo II - gastrite crônica (GC) com atividade inflamatória discreta (n = 101); Grupo III - CG com atividade inflamatória patente (n = 123); Grupo IV - pacientes com gastrite atrófica corpo (n = 20). Todos os cortes histológicos foram cuidadosamente avaliados por dois examinadores no aumento microscópico de imersão (1000×). Resultados: A identificação do Hp foi positiva pelo Giemsa e pela HE em, respectivamente, 111 (90,2%) e 93 (75,6%) pacientes do Grupo III (p < 0,01) e em 43 (42,6%) e 29 (28,7%) pacientes (p < 0,05) do Grupo II, e negativa nos grupos I e IV. Conclusão: Os resultados mostram que a coloração pelo Giemsa é superior à pelo HE para identificação do Hp em cortes histológicos. Conclui-se que o Hp pode ser visualizado pelo HE na maioria dos casos de GC, contudo, um número significativo de pacientes infectados pode ser negligenciado, independentemente da intensidade da resposta inflamatória. .

Effect of Isoliquiritigenin on C6 glioma cell proliferation and differentiation / 中国药理学通报

Aim To investigate the effects of isoliquiri-tigenin(ISL)on C6 glioma cell proliferation and differ-entiation.Methods C6 glioma cells’viability and proliferation were respectively measured by SRB test. Colony formation of C6 glioma cells from different groups was assayed.After culturing the cells from each group,giemsa staining was used to observe cell mor-phology.RT-PCR was applied to detect mRNA expres-sion of GFAP.Western blot was applied to detect the expression of GFAP.Results ISL effectively inhibited the viability of C6 glioma cells when compared with the control group in a concentration-dependent manner (P<0.01).The morphological observation under light mi-croscope showed that:in the control group,most of the undifferentiated C6 cells showed long fusiform and po-lygonal shape.Compared to the control group,the C6 cells treated with ISL revealed alteration in morphology such as astrocytes with smaller smooth,round body and much finer longer,tapering processes.The cloning for-mation rate detection revealed that:the colonies in the control group semerged earlier and were larger than those experimental ones,the cloning formation rate was higher,while almost no effective cells colony emerged in ISL treated groups(P <0.01 ).Western blot and RT-PCR analysis showed that GFAP expression in the ex-perimental groups increased(P <0.01).Conclusion ISL may inhibit the proliferation of C6 glioma cells and induce their differentiation.

Diagnoses of Giardia lamblia / 中国比较医学杂志

Objective Giardia lamblia is an important pathogen of zoonosis giardiasis , it poses a potential threat to the quality of SPF (specific pathogen-free) laboratory animals cannot be ignored.The aim of this study is to establish the method of rapid diagnosis of Giardia lamblia, and analyze the test results of 516 batches form 17 manufactures.Methods Direct microscopy, Giemsa-fast staining and multiplex polymerase chain reaction (multiplex PCR) were applied to detect Giardia lamblia.Results Numerous of Giardia lamblia trophozoites and cysts were detected in SPF laboratory animals by using direct microscopy and Giemsa-fast staining, and multiplex PCR were performed to identify 18S rDNA,β-giardin, TPI and GDH genes of DNA extracted from these trophozoites and cysts identified Giardia lamblia.Direct microscopy, Giemsa-fast staining, and multiplex PCR methods can be used to detect Giardia lamblia.Of the 2562 SPF laboratory animals studied, 22.9%(586/2562) were positive for Giardia lamblia.Conclusions Direct microscopy , Giemsa-fast staining , and multiplex PCR were effective techniques with high sensitivity and specificity for rapid diagnosis of Giardia lambliain.It is not satisfactory that the results of Giardia lamblia examination in 516 batches form 17 manufactures failed to meet the requirements 100%.