Distribution features of resident CD8 + T cells in human esophageal cancer tissues and its effects on the prognosis / 中华消化杂志

Objective:To explore the distribution features of resident CD8 + T cells infiltration in human esophageal cancer tissues and its clinical significance. Methods:Data from the Cancer Genome Atlas database were retrieved, the correlation between CD103 + CD8 + T cells and infiltration degree of conventional type 1 dendritic cell (cDC1), conventional type 2 dendritic cell (cDC2), type 3 dendritic cell(DC3) was investigated. From January 2006 to December 2008, 78 esophageal cancer tissues and 75 adjacent normal tissues from 78 esophageal cancer patients were collected by Shanghai Outdo Biotechnology Co., Ltd, the clinical data of patients was followed up by telephone until July 2015. The distribution of CD8 + T cells and CD103 + CD8 + T cells in cancer tissues and adjacent normal tissues was detected by multi-color labeling techniques and multispectral tissue imaging. The differences of the number and the ratio of CD8 + T cells and CD103 + CD8 + T cells in cancer tissues and adjacent normal tissues were compared. The Kaplan-Meier survival curves of patients with tissue infiltration of CD8 + T cells and CD103 + CD8 + T cells at different levels were drawn through the R language " survminer" package, and the best cut-off value was obtained. TNM stage, pathological stage and other clinical parameters of patients with high and low infiltration of CD8 + T cells, CD103 + CD8 + T cells were compared. Wilcoxon rank sum test, chi-square test, log-rank test and Cox proportional risk regression model statistical analysis were used to evaluate the prognostic value of the above indicators. Spearman correlation analysis was used for correlation analysis. Results:In the cancer tissues of patients with esophageal cancer, the infiltration degree of CD103 + CD8 + T cells was positively correlated with the infiltration degree of cDC1 cells, cDC2 cells and DC3 cells ( r=0.67, 0.53 and 0.47, all P<0.001). The percentage of CD8 + T cells in all cells in the whole tissue core of tumor tissues (63.09% (42.14%, 76.21%)) was higher than that of adjacent normal tissues (2.56% (1.68%, 5.38%)), and the difference was statistically significant ( U=41.00, P<0.001). The proportion of CD103 + CD8 + T cells in all cells in the whole tissue core of tumor tissues (7.92% (1.60%, 20.61%)) was higher than that of adjacent normal tissues (0.04% (0.01%, 0.10%)), and the difference was statistically significant ( U=857.50, P<0.001). The percentage of high CD8 + T cells infiltration in esophageal cancer tissues of patients with pathological stage Ⅰ+ Ⅱ was lower than that of patients with stage Ⅲ+ Ⅳ (57.9%, 33/57 vs. 85.7%, 18/21); the percentage of high CD103 + CD8 + T cells in CD8 + T cells in esophageal cancer tissues of patients with TNM stage Ⅰ+ Ⅱ was lower than that of patients with stage Ⅲ+ Ⅳ (21.6%, 8/37 vs. 48.8%, 20/41), and the differences were both statistically significant ( χ2=5.25 and 6.23, P=0.022 and 0.013). The results of Kaplan-Meier survival analysis and univariate Cox proportional risk regression model showed that the overall survival (OS) of patients with high CD8 + T cell infiltration was longer than that of patients with low CD8 + T cell infiltration ( HR=0.57, 95% confidence interval (95% CI) 0.34 to 0.96, P=0.034). There was no significant difference in OS between patients with high CD103 + CD8 + T cell infiltration and patients with low CD103 + CD8 + T cell infiltration ( HR=0.66, 95% CI 0.40 to 1.08, P>0.05). Conclusion:The high infiltration of CD103 + CD8 + T cells in esophageal cancer tissues are expected to be used as a prognostic predictor for patients with esophageal cancer, which is an important component of anti-tumor immune response in tumor microenvironment of esophageal cancer.

Desquamative Gingivitis as the First Clinical Sign of Pemphigus Vulgaris: A Case Report

ABSTRACT@#Desquamative gingivitis is characterised by desquamation of the gingiva with painful erosion and ulceration. It is predominantly a manifestation of several vesiculobullous diseases. Delayed diagnosis or misdiagnosis often led to disease progression. Pemphigus vulgaris is a chronic, life-threatening autoimmune disease resulting in blistering of the mucosa and skin. Oral lesions normally preceded skin lesions. Early diagnosis and treatment are important to prevent involvement of the skin, as the treatment and prognosis varies with extraoral involvement. Clinical, histopathological examination and direct immunofluorescent are necessary for the diagnosis of pemphigus vulgaris. Treatment of desquamative gingivitis involves improving oral hygiene, reduce irritation to the lesions and specific therapy to the underlying disease. This paper describes a case of a patient with desquamative gingivitis for one year, whom is ultimately diagnosed as having pemphigus vulgaris.

Seroprevalence, spatial analysis and risk factors of infection with Neospora caninum in cattle in Brazil's northern Pantanal wetland / Soroprevalência, análise espacial e fatores de risco da infecção por Neospora caninum em bovinos no Pantanal norte do Brasil

Abstract Neosporosis is caused by an obligate intracellular protozoan, Neospora caninum . It is considered one of the most widespread and frequent causes of abortion in cattle worldwide. To evaluate the prevalence of anti-N. caninum antibodies and associated risk factors, serum samples were collected from 2,452 bovines at 262 farms in the northern Pantanal, state of Mato Grosso, Brazil. Each farmer was asked to fill out a questionnaire for subsequent epidemiological data analysis. Anti-N. caninum antibodies were detected by means of the indirect immunofluorescent assay (IFA), using a cut-off dilution of 1:100. The overall anti-N. caninum antibodies prevalence was 25.44% (Confidence Interval - CI 95%; 20.10%; 30.78%), and the anti-N. caninum antibodies prevalence per herd was 76.72% (CI 95%; 71.60%; 81.84%). The presence of dogs, occurrence of abortion in cows, and sale of cattle for breeding were statistically associated with seropositivity in herds, while the risk of females being seropositive for N. caninum was higher in animals ≤ 6-years-old and in the presence of dogs. A spatial analysis indicated that the relative risk of the disease is spatially constant and that the farms with the highest prevalence of anti-N. caninum antibodies are located south of the region under study.

Resumo Neosporose é causada por um protozoário intracelular obrigatório, Neospora caninum. É considerada uma das causas mais comuns e frequentes de aborto em bovinos em todo o mundo. Para avaliar a prevalência de anticorpos anti- N. caninum e fatores de risco associados, amostras de soro foram coletadas de 2.452 bovinos em 262 fazendas no Pantanal norte, estado de Mato Grosso, Brasil. Cada fazendeiro preencheu um questionário para posterior análise dos dados epidemiológicos. Anticorpos anti-N. caninum foram detectados por meio da reação de imunofluorescência indireta (RIFI), utilizando um ponto de corte de 1:100. A prevalência total de anticorpos anti-N. caninum foi de 25,44% (Intervalo de Confiança - IC 95%; 20,10%; 30,78%) e a prevalência por rebanho foi de 76,72% (IC 95%; 71,60%; 81,84%). A presença de cães, a ocorrência de abortamento em vacas e a venda de bovinos para reprodução estiveram estatisticamente associadas à soropositividade em rebanhos, enquanto, o risco de fêmeas serem soropositivas para N. caninum foi maior em animais com idade ≤ 6 anos e na presença de cães. A análise espacial indicou que o risco relativo da doença é espacialmente constante e que as fazendas com maior prevalência de anticorpos anti-N. caninum estão localizados ao sul da região em estudo.

Diagnosis and treatment of graft-versus-host-disease after kidney transplantation: A Case Report and Literature Review / 中华器官移植杂志

Objective To investigate the diagnosis and treatment of graft-versus-host-disease (GVHD) after solid organ transplant (SOT) and the possible mechanism.Methods In this study,we retrospectively reported a rare case of GHVD after kidney transplantation and performed a literature review.This 51 year old male patient presented with over 10-day history of sporadic skin rash on postoperative day 50.Skin biopsy examination revealed GVHD.For further clear diagnosis,patient's peripheral blood was collected immediately for short tandem repeats (STR) enriched for CD3+ cells and we also performed immunofluorescent staining for patient's skin with specific HLA-A11 antibody,one of donor specific HLA sites.Meanwhile,the decreased immunosuppression was used for treatment.Results In our report,chimerism analysis by STR revealed no chimerism in patient's peripheral blood.However,immunofluorescent staining of patient's skin demonstrated abundant donor-derived lymphocytic infiltration existed.GVHD was definitely made in this case.After treatment with decreased immunosuppression and a low dose of methylprednisolone (MP),his clinical symptom was quickly alleviated.Now the patient was discharged and the renal function was normal.Conclusion Combined with literature review and our case report,we thought that chimerism analysis by STR and immunofluorescent staining by donor-specific HLA antibody were very useful for diagnosis of GVHD after SOT.Furthermore,GVHD referred to over-immunosuppression and prognosis of GVHD after kidney transplantation is usually satisfactory.

Seroprevalence, spatial analysis and risk factors of infection with Neospora caninum in cattle in Brazils northern Pantanal wetland

Abstract Neosporosis is caused by an obligate intracellular protozoan, Neospora caninum . It is considered one of the most widespread and frequent causes of abortion in cattle worldwide. To evaluate the prevalence of anti-N. caninum antibodies and associated risk factors, serum samples were collected from 2,452 bovines at 262 farms in the northern Pantanal, state of Mato Grosso, Brazil. Each farmer was asked to fill out a questionnaire for subsequent epidemiological data analysis. Anti-N. caninum antibodies were detected by means of the indirect immunofluorescent assay (IFA), using a cut-off dilution of 1:100. The overall anti-N. caninum antibodies prevalence was 25.44% (Confidence Interval - CI 95%; 20.10%; 30.78%), and the anti-N. caninum antibodies prevalence per herd was 76.72% (CI 95%; 71.60%; 81.84%). The presence of dogs, occurrence of abortion in cows, and sale of cattle for breeding were statistically associated with seropositivity in herds, while the risk of females being seropositive for N. caninum was higher in animals 6-years-old and in the presence of dogs. A spatial analysis indicated that the relative risk of the disease is spatially constant and that the farms with the highest prevalence of anti-N. caninum antibodies are located south of the region under study.

Resumo Neosporose é causada por um protozoário intracelular obrigatório, Neospora caninum. É considerada uma das causas mais comuns e frequentes de aborto em bovinos em todo o mundo. Para avaliar a prevalência de anticorpos anti- N. caninum e fatores de risco associados, amostras de soro foram coletadas de 2.452 bovinos em 262 fazendas no Pantanal norte, estado de Mato Grosso, Brasil. Cada fazendeiro preencheu um questionário para posterior análise dos dados epidemiológicos. Anticorpos anti-N. caninum foram detectados por meio da reação de imunofluorescência indireta (RIFI), utilizando um ponto de corte de 1:100. A prevalência total de anticorpos anti-N. caninum foi de 25,44% (Intervalo de Confiança - IC 95%; 20,10%; 30,78%) e a prevalência por rebanho foi de 76,72% (IC 95%; 71,60%; 81,84%). A presença de cães, a ocorrência de abortamento em vacas e a venda de bovinos para reprodução estiveram estatisticamente associadas à soropositividade em rebanhos, enquanto, o risco de fêmeas serem soropositivas para N. caninum foi maior em animais com idade 6 anos e na presença de cães. A análise espacial indicou que o risco relativo da doença é espacialmente constante e que as fazendas com maior prevalência de anticorpos anti-N. caninum estão localizados ao sul da região em estudo.

Preparation of VEGFRⅡ-mediated Targeted Microbubble Image Agent and its Cancer Targeting Ability in vitro / 中国药师

Objective:To prepare vascular endothelial growth factor receptorⅡ ( VEGFRⅡ)-mediated targeted sulphur hexafluo-ride microbubble image agent and evaluate its targeting ability in human breast cancer MCF-7 in vitro. Methods:The sulphur hexafluo-ride microbubble image agent was prepared by a membrane dispersion-ultrasonic method and optimized by Box-Behnken design using design-expert 8. 0. 6 software. VEGFRⅡ-mediated targeted microbubble image agent was prepared by an absorption method. VEGFRⅡ-mediated targeted microbubble image agent was characterized by the morphology, particle size distribution and zeta potential. The targeting ability of the image agent in human breast cancer MCF-7 in vitro was detected by an immunofluorescent assay. Results:The particle size, polydispersion index and zeta potential of VEGFRⅡ-mediated targeted microbubble image agent was (3. 81 ± 0. 32) μm, 0. 261 ± 0. 037 and ( -25. 7 ± 2. 8) mV, respectively. The microbubbles were small and spherical with smooth surface as seen under a transmission electron microscope. VEGFRⅡ-mediated targeted microbubble image agent could combine with human breast cancer MCF-7 specifically in vitro. Conclusion: The absorption preparation technology of the targeted microbubble image agent is feasible, and the microbubble image agent has strong and especial targeting function in vitro.

Immunofluorescence probe based on fluorescent carbon dots and its application to the special recognition of Escherichia coli / 军事医学

Objective Carbon dots (CDs) are an emerging carbon nano-material which is environmentally-friendly, economical , efficient and stable .Their fluorescence properties can match the semiconductor quantum dot .Moreover , CDs have more excellent biocompatibilities .The purpose of this experiment is to apply CDs to the fluorescent immune probe to make them a new label , which can replace the traditional fluorescent dyes .Methods Using microwave heating method , the high strength fluorescent carbon dots were prepared .Wtih the EDC coupling method , the high strength fluorescent car-bon dots could bond with Escherichia coli antibodies to form a complex immune fluorescent probe .Specific recognition exper-iments were carried out in the model of E.coli O157∶H7.Results CDs were successfully applied to immune recognition of E.coli O157∶H7 and multicolor fluorescence was observed .Conclusion CDs can serve as a label of the fluorescent im-mune probe , and are expected to become a new type of low toxicity biosensor with independent intellectual property rights .

Frequency of Toxoplasma gondii antibodies in bovines in the state of Pernambuco, Brazil / Frequência de anticorpos anti-Toxoplasma gondii em bovinos do estado de Pernambuco, Brasil

Toxoplasmosis is a parasitic disease caused by Toxoplasma gondii that affects homeothermic animals, including humans. Felines are considered the definitive host of this parasite, while other animals act as intermediate hosts. The purpose of this study was to assess the frequency of anti-T. gondii IgG antibodies in bovines in the state of Pernambuco, Brazil. Serum samples (n = 427) from animals in 13 municipalities of the coastal forest/plantation region of the state were analyzed using the immunofluorescent antibody test (IFAT). The overall results revealed a prevalence rate of 16.63% (27/427). High percentages of positivity were found among animals aged 25 to 36 months (28.57%; 30/42) and in males (22.22%; 2/9). The present findings suggest that bovine toxoplasmosis is endemic in the area under study.

Toxoplasmose é uma doença parasitária que infecta todos os animais homeotérmicos, incluindo o homem. Na epidemiologia da infecção, os felinos são os hospedeiros definitivos, e outros animais são os hospedeiros intermediários do Toxoplasma gondii. O objetivo deste trabalho foi determinar a frequência de anticorpos IgG anti-Toxoplasma gondii em bovinos do Estado de Pernambuco. Para tanto, foram coletadas 427 amostras de soro sanguíneo de bovinos provenientes de 13 municípios localizados na Zona da Mata do Estado de Pernambuco. Tais amostras foram analisadas pela técnica de Imunofluorescência Indireta para pesquisa de anticorpos anti-Toxoplasma gondii. A frequência observada foi de 16,63% (27/427). Anticorpos anti-Toxoplasma gondii foram mais frequentes (28,57%) em animais com idade entre 25-36 meses de idade. Considerando-se por sexo, observou-se uma maior positividade entre os machos que apresentaram 22,22% (2/9). Os resultados demonstram que a toxoplasmose é endêmica nessa região.

Development of the mouse spinal cord and neuroapoptosis / 解剖学报

Objective To investigate the neural proliferation , differentiation and apoptosis of the developing spinal cord of the mouse and to discuss the mechanism of spinal cord ’ s development .Methods 5-Bromodeoxyuridine ( BrdU) assay was used to mark the proliferative neural stem cells , and the immunofluorescent stainings ( DCX, NeuN and Caspase8) were carried out to visualize the newborn neurons , mature cells and apoptotic cells in the spinal cord with 173 mice arrange from E18 to P90.Results BrdU positive neural stem cells appeared evenly in the spinal cord at early days . With age increasing , the neural stem cells differentiated into neuroglial cells and neurons .The newborn neurons in the subventricular zone migrated toward the intermediate zone ( putative gray matter ) and differentiated into mature neurons gradually .With neurons ’ concentrating towards the center , the gray matter formed an “H” shape .In the meantime , with neural differentiation , some apoptotic neurons appeared among the newborn neurons and mature neurons . Double immunostaining showed that most apoptotic neurons were newborn neurons , suggesting the neuroapoptosis more likely occurred in newborn neurons .The statistical data showed that the number of DCX , NeuN and Caspase-8 positive cells reduced with age increasing , suggesting neural differentiation and neuroapoptosis decreased during spinal cord ’ s development .Conclusion Neural proliferation , neural differentiation and neuroapoptosis occur in developing spinal cord . They work together to regulate the formation and development of the spinal cord .

Effects of Jagged1 on hippocampal radial glial cells’ proliferation and neuronal differentiation / 解剖学报

Objective To investigate the effect of Jagged1 on hippocampal radial glial cells (RGCs) proliferation and neuronal differentiation in vitro.Methods Hippocampal RGCs were cultured in vitro, the agonist Jagged1 and(or) inhibitor DAPT of Notch signaling were added into the culture medium , and then the cells were divided into control group , Jagged1 group, Jagged1 combined with DAPT group and DAPT group .CCK-8 regent was used to detect cells ’ vitality;immunofluorescent was used to detect the number of BLBP /Ki67 double labeled cells and differentiated microtubule associated protein-2(MAP-2) positive cells.Results Cell vitality in Jagged1 group was obviously higher than that of the other groups .The number of BLBP/Ki67 double labeled cells and differentiated MAP-2 positive cells were more than other groups.Conclusion Jagged1 promotes the proliferation and neuronal differentiation of hippocampal RGCs in vitro.

Investigation on Ehrlichia chaffeensis and Anaplasma phagocytophilum infection among farmers and domestic animals in rural areas of Beijing, China / 中华流行病学杂志

Objective To investigate the status of Ehrlichia (E.)chaffeensis and A naplasma (A.) phagocytophilum infection among farming populations and domestic animals in the rural area of Beijing,China.Methods Blood samples from 562 farmers and 163 blood samples including 90 goats,71 ox and 2 dogs,were collected.Specificity of IgG antibodies against E.chaffeensis and A.phagocytophilum were tested by micro-indirect immunofluorescent assay (mIFA).16S rRNA genes of A.phagocytophilum were amplified from the domestic animal blood samples and products were sequenced and analyzed by nested PCR.Results The positive rates of E.chaffeensis and A.phagocytophilum antibody were 16.5％ and 14.0％ respectively for farmers.The total positive rates of A.phagocytophilum were 2.3％ and 0 for both goats and oxen respectively.No antibody was found for the 2 tested dogs.The PCR positive rates were 48.9％ and 23.9％ for goats and oxen respectively.Three dominant varieties of A.phagocytophilum were demonstrated in goats and oxen.Conclusion The prevalence rates of E.chaffeensis and A.phagocytophilum were identified in the rural areas of Beijing.

IgG Avidity ELISA Test for Diagnosis of Acute Toxoplasmosis in Humans

Serum samples, 100 in the total number, were collected from different laboratories in Tehran, Iran and tested for anti-Toxoplasma specific IgG and IgM antibodies using indirect immunofluorescent antibody test (IFAT). Using the IgG (chronic) and IgM (acute) positive samples, the IgG avidity test was performed by ELISA in duplicate rows of 96-well microtiter plates. One row was washed with 6 M urea and the other with PBS (pH 7.2), then the avidity index (AI) was calculated. Sixteen out of 18 (88.9%) sera with acute toxoplasmosis showed low avidity levels (AI60). Six sera had borderline ranges of AI. The results showed that the IgG avidity test by ELISA could distinguish the acute and chronic stages of toxoplasmosis in humans.

IgG Avidity ELISA Test for Diagnosis of Acute Toxoplasmosis in Humans

Serum samples, 100 in the total number, were collected from different laboratories in Tehran, Iran and tested for anti-Toxoplasma specific IgG and IgM antibodies using indirect immunofluorescent antibody test (IFAT). Using the IgG (chronic) and IgM (acute) positive samples, the IgG avidity test was performed by ELISA in duplicate rows of 96-well microtiter plates. One row was washed with 6 M urea and the other with PBS (pH 7.2), then the avidity index (AI) was calculated. Sixteen out of 18 (88.9%) sera with acute toxoplasmosis showed low avidity levels (AI60). Six sera had borderline ranges of AI. The results showed that the IgG avidity test by ELISA could distinguish the acute and chronic stages of toxoplasmosis in humans.

A sensitive and specific ES-31 antigen detection based fluorometric assay for confirmation of Mycobacterium tuberculosis in cell culture.

Confirmation of presence of M. tuberculosis bacilli on microscopic examination is very important in diagnosis of tuberculosis. The present study was undertaken to find the usefulness of mycobacterial ES-31 serine protease as a marker to detect tuberculosis bacilli using fluorescein isothiocyanate conjugated anti-ES-31 serine protease antibody. This immunofluorescence method was compared with Ziehl-Neelsen and auramine-O staining methods for detection of tuberculosis bacilli. Slides were prepared for each serially diluted tuberculosis H37Ra bacilli (1×107 bacilli/ml to 5 bacilli/ml). Slides for each dilution group were stained by ZN method, auramine-O and immunostaining methods using fluorescein isothiocyanate conjugated anti-ES-31 serine protease antibody. ZN staining method showed efficacy for detection of M. tuberculosis H37Ra upto 1×104 bacilli/ml while auramine-O method showed upto 1×102 bacilli/ml. The presence of bacilli was indicated by green fluorescence on immunostaining using anti-ES-31 antibody conjugate and this method was effective upto 10 bacilli/ml. The slides which were negative for ZN (1×103 cells/ml) and auramine-O (100 cells/ml) method showed positivity on restaining with immunofluorescent staining method. The results of this preliminary study showed that immunofluorescent staining method using specific anti-ES-31 antibody conjugate was more sensitive for detection of tuberculosis bacilli than ZN and auramine-O methods in samples of laboratory strain. The utility of this method will be studied further in clinical specimens.

Detecção de anticorpos anti-Leishmania infantum syn chagasi em cães de São José do Rio Preto, São Paulo / Anti-Leishmania infantum syn chagasi antibodies in dogs from São José do Rio Preto, São paulo

A leishmaniose visceral (LV) é uma zoonose que vem se expandido por todo o território paulista desde 1998, quando foi identificado o primeiro caso canino autóctone, no município de Araçatuba. O objetivo do presente estudo foi determinar a ocorrência de anticorpos anti-Leishmania infantum syn chagasi em amostras de soro de 584 cães de São José do Rio Preto, São Paulo, área não endêmica para a doença. Cinco cães (0,86%) foram soro reagentes pela técnica de ELISA e um (0,17%) por imunocromatografia. A reação de imunofluorescência indireta, realizada em 138 animais que possuíam densidades ópticas acima ou próximas ao ponto de corte do ELISA evidenciou dois cães (1,45%) com títulos acima de 1:40. Apenas um animal foi sororeagente nas três técnicas sorológicas. Apesar deste cão não apresentar histórico de deslocamento para áreas endêmicas, havia sido adquirido em região com casos caninos e humanos de LV. Os resultados obtidos no presente estudo fazem supor que não existiam casos autóctones de LV canina na população estudada.

Visceral leishmaniasis (VL) has been a widespread zoonosis in São Paulo since 1998, when the first autochthonous canine case was identified in Araçatuba. The aim of this study was to determine the occurrence of anti–Leishmania infantum syn chagasi antibodies in serum samples of 584 dogs from São José do Rio Preto, São Paulo, a non endemic area for the disease. Five dogs (0.86%) seroconverted by ELISA and one (0.17%) by immunochromatography. The indirect immunofluorescent reaction, carried out in 138 animals whose optical densities were above or close to ELISA’s cutt-off point, evidenced two dogs (1.45%) with titers above 1:40. Only one dog was serum-reactive on the three techniques. Although there was not a history of displacing this animal to endemic areas, the dog had been acquired in a region with canine and human cases of VL. These results suggests that there were no autochthonous cases of canine VL in this population.

Modulation of TNF on the expression of protease activated receptors in P815 mast mells / 中华微生物学和免疫学杂志

Objective To investigate the potential influence of TNF on the expression of protease activated receptor (PAR)-1,2,3 and 4 by using P815 mast cells. Methods After being challenged with various concentrations of TNF for 2 h, 6 h and 16 h, the P815 mast cells were treated with or without Triton X-100 and the PAR expressions were detected by flow cytometry and immunofluorescent microscopy. Results Compared with the corresponding controls, TNF concentration-dependently upregulated expressions of PAR-2 and PAR-4 both in Triton X-100-treated and the untreated groups, but had no significant effect on the expression of PAR-1 and PAR-3. Moreover, no significantly different expressions of TNF-induced PAR-1, 2, 3 were observed between Triton X-100-treated and the untreated groups, whereas Triton X-100-treated PAR-4 expressions were significantly enhanced by TNF compared with the Triton X-100-untreated ones. Conclusion TNF can up-regulate PAR-2, 4 expression of P815 mast cells but has little effect on the expression of PAR-1, 3 correspondingly. And Triton X-100 treatment had no significant effect on TNF-modulated expression of PARs in P815 mast cells.

Analysis of etiological agent in 771 children with lower respiratory infection / 国际儿科学杂志

Objective To detect and analyze viral pathogens and the etiological agent distribution of hospitalized children with lower respiratory infection. Methods We adopted direct immunofluorescence to detect the seven frequent respirovirus antigens from nasopharyngeal secretion in 771 hospitalized patients with lower respiratory infection.Results Virus detection of 243 cases was positive in 771, the total positives ratio was 31.52%. Respirovirus infection had obviously relation with age and season. The positive rate of virus infection was highest in asthma, followed in bronchiolilis and asthmatic bronchitis, which were 58.80% and 35.56% respectively. Conclusion Different ages and different seasons presented different viral infection and different virus types. Infection rate in infant was higher obviously than that in the other ages, respiratovy syncytial virus and parain- fluenza virus were the major virus in lower respiratory infection. Viral infection was important reason to cause children asthma.

3º Consenso Brasileiro para pesquisa de autoanticorpos em células HEp-2 (FAN): recomendações para padronização do ensaio de pesquisa de autoanticorpos em células HEp-2, controle de qualidade e associações clínicas / Third Brazilian Consensus for autoantibodies screening in HEp-2 cells (ANA): recommendations for standardization of autoantibodies screening trial in HEp-2 cells, quality control and clinical associations

OBJETIVO: O 3º Consenso Brasileiro para pesquisa de autoanticorpos em Células HEp-2 (FAN) teve como propósito avaliar as dificuldades de implantação do 2º Consenso ocorrido no ano de 2002, discutir estratégias para controlar a qualidade do ensaio e promover a atualização das associações clínicas dos diversos padrões. MÉTODOS: Participaram do encontro em Goiânia nos dias 13 e 14 de abril de 2008 pesquisadores e especialistas de diversos centros universitários e laboratórios clínicos de diferentes regiões do Brasil, com o propósito de discutir e aprovar as recomendações que visam à melhor padronização, interpretação e utilização do ensaio pelos clínicos. Representantes comerciais de diferentes empresas produtoras de insumos para realização do teste de FAN foram convidados como ouvintes. RESULTADOS E CONCLUSÕES: O 3º Consenso enfatizou a necessidade do controle de qualidade em imunofluorescência dada a heterogeneidade de microscópios e reagentes disponíveis no mercado, promoveu adequações na terminologia utilizada para classificar os diferentes padrões e, finalmente, atualizou as associações clínicas com finalidade de facilitar cada vez mais o melhor uso do ensaio pelos clínicos.

OBJECTIVE: The Third Brazilian Consensus for autoantibodies Screening in HEp-2 cells had as purpose the evaluation of difficulties in the accomplishment of the 2nd Consensus recommendations that took place in the year of 2002, the discussion of strategies for quality control of the assay and the promotion of an update of the clinical associations of the several immunofluorescent patterns. METHODS: Several ANA experts from university centers and private laboratories in different areas in Brazil joined the workshop in Goiânia on 2008 April 13 and 14 with the purpose of discussing and approving the recommendations for standardization, interpretation and use of the test by physicians. Commercial representatives of different ANA slide brands were also invited as listeners to the workshop. RESULTS AND CONCLUSIONS: The 3rd Consensus emphasized the need for quality control in indirect immunofluorescent since there is a considerable heterogeneity of available microscopes and reagents. It also promoted adaptations in the previously approved terminology used to classify the different patterns and finally updated the clinical associations of the several patterns with the purpose of providing guidance for interpretation of the assay by clinical pathologists and assistant physicians.

Experiência da adoção do I e II Consensos Brasileiros de Fator Antinuclear por Imunofluorescência Indireta em Células HEp-2 em um hospital universitário / Experience of an university hospital on the implementation of I and II Brazilian Consensuses for Standardization of ANA in HEp-2 Cells

OBJETIVO: Analisar a prevalência, padrões e títulos do Fator Antinuclear (FAN) por imunofluorescência indireta (IFI) em células HEp-2 em um hospital universitário após a adoção do I e II Consensos Nacional para Padronização dos Laudos de FAN em Células HEp-2. MÉTODO: Foi realizado um estudo transversal, em que foram revisados os laudos de FAN por IFI originários de solicitações encaminhadas ao Serviço de Patologia Clínica do Hospital de Clínicas de Porto Alegre (SPC/HCPA) entre 2002 e 2005. RESULTADOS: Foram analisados 12.095 testes de FAN no período entre 2002 e 2005. As solicitações com resultado reagente foram de 2.577 (21,30 por cento), com média anual de 644±233). Houve um aumento significativo na proporção de resultados reagentes posterior à adoção dos Consensos (p < 0,001). A Reumatologia foi a especialidade que mais solicitou exames por paciente atendido, mas houve um declínio nesse número nos anos posteriores à adoção do Consenso, ocorrido em 2004 (p < 0,001). O padrão de imunofluorescência de FAN mais frequentemente encontrado foi o padrão nuclear pontilhado fino, presente em 52,3 por cento dos resultados reagentes (453/866), e os títulos mais encontrados foram 1/80 e 1/160 (27,8 por cento e 29,4 por cento, respectivamente). CONCLUSÃO: Após a adoção do Consenso Nacional de Padronização de Laudos de FAN, observou-se um aumento de exames com resultados reagentes, na sua maioria, com títulos baixos e padrão nuclear pontilhado fino. Na Reumatologia, observou-se uma diminuição no número de solicitações desse exame. As potenciais causas para essas observações são discutidas, mas seu real impacto sobre a situação clínica do paciente e seu tratamento merece ser mais bem estudado.

OBJECTIVE: To evaluate the prevalence of patterns and titers of antinuclear antibodies (ANA) detected by indirect immunofluorescence (IIF) technique on HEp-2 cells in a university hospital following the introduction of I and II Brazilian Consensuses for Standardization of ANA in HEp-2 Cells. METHODS:A transversal study was performed between 2002 and 2005 during which all ANA orders to Serviço de Hospital de Clínicas de Porto Alegre (SPC/HCPA) and cognate results were reviewed. RESULTS:12.095 tests of ANA were revised. The number of positive results during this period was 2.577 (21.30 percent), annual mean 644 (SD: 233). A marked increase in the number of positive results was observed following the introduction of the Consensuses (p < 0.001). Rheumatology was the medical specialty which requested the highest number of ANA testing per patient although a significant decrease of these numbers was observed after the introduction of the Consensus in 2004 (p < 0,001). Nuclear fine speckled immunofluorescence labeling was the most frequently ANA pattern observed, 52.3 percent (453/866), and low ANA titers (1/80 and 1/160) more commonly detected (27.8 percent and 29.4 percent, respectively). CONCLUSION: Following the introduction of the Brazilian Consensus for standardization of ANA in HEp-2 cells an increased number of positive results was observed, mostly in low titers and with nuclear fine speckled immunofluorescence pattern. Moreover, there were decreasing numbers of ANA orders by rheumatologists in the same period. Potential causes for these observations are discussed but the real impact in the clinical condition of the patient and therapy deserves to be better studied.

Expression and Distribution of BDNF (Brain Derived Neurotrophic Factor) in the Rat Hypothalamus / 대한해부학회지

BDNF belongs to the neurotrophin family and important molecular mediator of functional and structural plasticity. The highest levels of BDNF are found in the hippocampus and hypothalamus of the adult rat. Hypothalamus is important because of its high degree of plasticity, but little is known about distribution of BDNF in hypothalamic nuclei. Therefore, it is necessary to study distribution and expression pattern of BDNF in each hypothalamic nuclei to understand changes of BDNF through various neural damages including spinal cord injury. Through this experiment, we found specific BDNF expression pattern in some regions of hypothalamus and the results are as follows. 1) BDNF expressions were found in median eminence, arcuate nucleus, supraoptic nucleus, and periventricular nucleus of rat hypothalamus. 2) BDNF immunoreactive cells and nerve fibers were of various shapes and sizes. 3) Glial cells also express BDNF in certain hypothalamic nuclei. These results seem to be useful for future investigations of neurochemical changes in the hypothalamus induced by various neural trauma or degenerative changes