Habilidades procedimentales invariantes para el diagnóstico y el tratamiento de la peritonitis posoperatoria

Introducción: Las habilidades procedimentales invariantes en las peritonitis posoperatorias, les permite a los cirujanos el diagnóstico temprano y la reoperación abdominal urgente. Objetivo: Diseñar las habilidades procedimentales invariantes de atención diagnóstica y terapéutica a la peritonitis posoperatoria dirigida a los cirujanos. Método: Se realizó una investigación cualitativa en el Hospital Clínico Quirúrgico "Ambrosio Grillo" de Santiago de Cuba en el trimestre mayo-julio de 2022. La población de estudio estuvo constituida por 33 profesionales tratantes de la peritonitis posoperatoria: 22 médicos (12 cirujanos y 10 intensivistas) y 11 licenciadas en Enfermería. Se utilizó la sistematización como método teórico y como método empírico la revisión documental con la revisión de las historias clínicas y la observación con la práctica atencional-docente de la autora principal. Resultados: Se diseñaron las habilidades procedimentales invariantes para el diagnóstico temprano y la terapéutica de la peritonitis posoperatoria dirigida a los cirujanos con especial importancia en el método clínico. La autopreparación del tema en cuestión posibilita la complementación de conocimientos y habilidades aprendidas, así como la actualización de saberes que garantiza la atención clínica quirúrgica a este complejo enfermo. Conclusiones: El diseño de las habilidades procedimentales invariantes para el diagnóstico temprano y la terapéutica de la peritonitis posoperatoria en la necesaria superación permanente y continuada de los cirujanos representa una herramienta asistencial-docente que contribuye al mejoramiento del desempeño profesional ante este tipo de enfermo.

Introduction: Invariant procedural skills in postoperative peritonitis allow surgeons an early diagnosis and the performance of urgent abdominal resurgery. Objective: To design invariant procedural skills to be implemented for surgeons on the diagnostic and therapeutic care of postoperative peritonitis. Method: A qualitative research was conducted at the Hospital Clínico Quirúrgico "Ambrosio Grillo" of Santiago de Cuba in the quarter period of May-July 2022. The study population consisted of 33 specialists on postoperative peritonitis: 22 physicians (12 surgeons and 10 intensivists) and 11 graduated bachelor in nursing. Systematization was used as the theoretical method and the empirical methods used were the document analysis supported with the review of clinical histories and the observation with the main author's attentional-teaching practice. Results: It was designed an invariant procedural skills for the early diagnosis and therapy of postoperative peritonitis with special emphasis on the clinical method. The self-preparation on the subject makes possible the complementation of both knowledge and skills learned, as well as an update of knowledge that guarantees the surgical clinical care of these complex patients. Conclusions: The design of invariant procedural skills for the early diagnosis and therapy of postoperative peritonitis in the necessary permanent and continuous knowledge improvement of surgeons represents a care-teaching tool that contributes to improve the professional performance at the time to attend to this type of patient.

Introdução: As habilidades processuais invariantes na peritonite pós-operatória permitem aos cirurgiões o diagnóstico precoce e a reoperação abdominal urgente. Objetivo: Projetar as habilidades processuais invariantes de atenção diagnóstica e terapêutica para peritonite pós-operatória destinadas a cirurgiões. Método: Uma pesquisa qualitativa foi realizada no Hospital Clínico Cirúrgico "Ambrosio Grillo" em Santiago de Cuba no trimestre maio-julho de 2022. A população do estudo consistiu em 33 profissionais que tratam de peritonite pós-operatória: 22 médicos (12 cirurgiões e 10 intensivistas) e 11 graduados em enfermagem. Utilizou-se como método teórico a sistematização e como métodos empíricos a revição documental com revisão de histórias clínicas e a observação com a prática docente-atencional da autora principal. Resultados: Desenharam-se competências processuais invariantes para o diagnóstico precoce e terapêutica da peritonite pós-operatória dirigidas a cirurgiões com especial importância no método clínico. O autopreparo do sujeito em questão possibilita a complementação dos conhecimentos e habilidades aprendidas, bem como a atualização de conhecimentos que garantem o cuidado clínico cirúrgico desta complexa doença. Conclusões: O desenho de habilidades processuais invariantes para o diagnóstico precoce e tratamento da peritonite pós-operatória no necessário aperfeiçoamento permanente e contínuo dos cirurgiões representa uma ferramenta cuidado-ensino que contribui para o aperfeiçoamento da atuação profissional frente a este tipo de paciente.

Immune responses of mucosal-associated invariant T cells in infectious diseases / 中华微生物学和免疫学杂志

Mucosal-associated invariant T (MAIT) cells are highly conserved immune cells that could participate in innate and adaptive immune responses after being activated by major histocompatibility complex class 1-related molecule (MR1) pathway or cytokine pathway. At present, it has been confirmed that a large number of MAIT cells exist in human peripheral blood and specific tissues, and play an important role in infectious diseases. This review focused on the role of MAIT cells in immune responses to different pathogens. Additionally, the therapeutic methods and challenges of targeting MAIT cells in infectious diseases were also discussed.

Changes in the frequency, phenotype, and function of mucosal-associated invariant T lymphocytes in peripheral blood of children with metabolic associated fatty liver disease / 临床肝胆病杂志

Objective To investigate the change in mucosal-associated invariant T (MAIT) lymphocytes in peripheral blood of children with metabolic associated fatty liver disease (MAFLD) and its clinical significance. Methods A total of 18 children with MAFLD who attended Hunan Children's Hospital from March to May, 2022, were enrolled as MAFLD group, and 20 normal children who attended the hospital during the same period of time were enrolled as control group. Peripheral blood samples were collected, and flow cytometry was used to measure the percentages of MAIT lymphocytes (CD3 + CDl61 + TCRVα7.2 + cells), different MAIT lymphocyte subsets (CD4 + CD8 - MAIT cells, CD4 - CD8 - MAIT lymphocytes, CD4 - CD8 + MAIT lymphocytes, and CD4 + CD8 + MAIT lymphocytes), and MAIT lymphocytes expressing PD-1, CD69, perforin, CD107α, CXCR3, CXCR6, and CCR6. The correlation of MAIT lymphocyte frequency with liver inflammation, fat content, and fibrosis degree was analyzed. The t -test was used for comparison of normally distributed continuous data between two groups, and the Mann-Whitney U test was used for comparison of non-normally distributed continuous data between two groups. The Spearman correlation analysis was used for correlation analysis. Results Compared with the control group, the MAFLD group had significant increases in the percentage of MAIT lymphocytes in peripheral blood, the percentages of MAIT cells expressing PD-1, CD69, CD107α, CXCR3, CXCR6 and CCR6, and the percentages of CD4 + CD8 - MAIT cells and CD4 + CD8 + MAIT lymphocytes among MAIT cell subsets (all P < 0.05), as well as a significant reduction in the percentage of CD4 - CD8 + MAIT cells among MAIT cell subsets ( P < 0.001). The percentages of CD4 + CD8 + MAIT lymphocytes and CD107α-positive MAIT lymphocytes were negatively correlated with alanine aminotransferase ( r =-0.474 and -0.550, P =0.047 and 0.018). Conclusion The migration of peripheral blood MAIT lymphocytes to the liver plays a protective role against liver inflammation in children with MAFLD.

Invariant Natural Killer T-cells and their subtypes may play a role in the pathogenesis of endometriosis

Abstract Objective To evaluate the frequencies of iNKT cells and their subsets in patients with deep endometriosis. Methods A case-control study was conducted between 2013 and 2015, with 73 patients distributed into two groups: 47 women with a histological diagnosis of endometriosis and 26 controls. Peripheral blood, endometriosis lesions, and healthy peritoneal samples were collected on the day of surgery to determine the frequencies of iNKT cells and subtypes via flow cytometry analysis. Results The authors observed a lower number of iNKT (p= 0.01) and Double-Negative (DN) iNKT cells (p= 0.02) in the blood of patients with endometriosis than in the control group. The number of DN iNKT IL-17+ cells in the secretory phase was lower in the endometriosis group (p= 0.049). There was an increase in the secretion of IL-17 by CD4+ iNKT cells in the blood of patients with endometriosis and severe dysmenorrhea (p= 0.038), and severe acyclic pelvic pain (p= 0.048). Patients with severe dysmenorrhea also had a decreased number of CD4+ CCR7+ cells (p= 0.022). Conclusion The decreased number of total iNKT and DN iNKT cells in patients with endometriosis suggests that iNKT cells play a role in the pathogenesis of endometriosis and can be used to develop new diagnostic and therapeutic agents.

Mucosal-associated invariant T cells expression and its clinical significance in ankylosing spondylitis / 中华风湿病学杂志

Objective:To investigate the expression of Mucosal-associated invariant T (MAIT) cells in the peripheral blood of ankylosing spondylitis (AS) patients, and to analyze its clinical significance.Methods:Sixty-four AS patients from September 2020 to September 2022 in Fujian Provincial Hospital and 60 healthy controls were enrolled. The expression of MAIT cells and CD69 +MAIT cells were detected by immunofluorescence. Spondylarthritis research consortium of Canada (SPARCC) scores of sacroiliac joint magnetic resonance imaging (MRI) were calculated and were analyzed based on clinical data. The changes of MAIT cells were observed in 7 patients with AS after treatment. t test, variance analysis and Pearson correlation analysis were used for statistical analysis. Results:The expression of MAIT cells in the peripheral of AS patients was significantly lower than that in the controls [(2.81±0.27)% and (4.46±0.86)% respectively, t=2.33, P=0.022], while CD69 +MAIT cells were significantly higher [(12.0±1.0)% and (7.8±1.2)% respectively, t=2.31, P=0.024]. The level of CD69 +MAIT cells were significantly positively correlated with erythrocyte sedimentation rate (ESR) ( r=0.39, P=0.010), C-reactive protein (CRP) ( r=0.36, P=0.012), ASDAS ( r=0.35, P=0.013) and SPARCC scores ( r=0.38, P=0.006) of AS patients. ASDAS scores and CD69 +MAIT cells obviously decreased in 7 treated AS patients ( F=7.62, P=0.007 and F=4.97, P=0.027 respectively). Conclusion:The expression of peripheral MAIT cells in AS patients is lower than that in healthy controls, but the number of the activated MAIT cells is increased. Levels of activated MAIT cells may in some extent reflect the inflammatory state and disease activity of AS, as well as inflammatory destruction of sacroiliac joint, and therapeutic effect. MAIT cells may partially participate in the inflammatory response and play a role in the pathogenesis of AS.

The mechanism of iNKT2 cell improving lipid deposition in the liver of nonalcoholic fatty liver mice / 中华内分泌代谢杂志

Objective:To investigate the mechanism of invariant natural killer T (iNKT)2 cell improving hepatic fat deposition in nonalcoholic fatty liver (NAFL).Methods:NAFL model was established by feeding C57BL/6J mice with high fat diet. The levels of serum total cholesterol, triglyceride, high density lipoprotein-cholesterol (HDL-C), low density lipoprotein-cholesterol (LDL-C), alanine aminotransferase (ALT), and aspartate aminotransferase (AST) in the peripheral blood of mice were analyzed using automatic biochemical analyzer. The pathological changes of liver were observed with HE staining. The cell frequencies of iNKT, iNKT1, and iNKT2 in liver were detected by flow cytometry. Western blotting was used to detect the expression of sterol regulatory element binding protein 1c (SREBP-1c), peroxisome proliferator activated receptor (PPAR)-α, and nuclear factor-κB (NF-κB) in liver tissues.Results:Compared with control group, the body weight of NAFL mice increased, the levels of total cholesterol, HDL-C, LDL-C, ALT, and liver fat deposition increased, the protein expression of SREBP-1c and PPAR-α in liver increased as well as the the protein phosphorylation level of NF-κB. After intraperitoneal injection of α-galactosylceramide (α-GalCer), the levels of total cholesterol, HDL-C, and LDL-C, liver fat deposition decreased, liver SREBP-1c was down-regulated, PPAR-α expression was up-regulated, and the proportion of liver iNKT2 subgroup increased in NAFL mice.Conclusion:iNKT2 cells improve NAFL liver fat deposition, which is related to the down-regulation of SREBP-1c and up-regulation of PPAR-α.

Predictive Role of Circulating Immune Cell Subtypes Early after Allogeneic Hematopoietic Stem Cell Transplantation in Patients with Acute Leukemia

BACKGROUND AND OBJECTIVES: Cells of innate immunity normally recover in the first weeks to months after allogenenic hematopoietic stem cell transplantation (allo-HSCT). Their relevance in terms of graft-versus-host disease (GVHD) and graft-versus-leukemia (GVL) effect is largely unknown. The predictive role of early recovery in the immune cells on acute GVHD and GVL effect after allo-HSCT was investigated in patients with acute leukemia who achieved the first complete remission. METHODS: Peripheral blood samples were taken at the median of 14 days (range, 12~29 days) after allo-HSCT. A cohort including 119 samples and characteristics of patients were analyzed. Immune cell populations were identified by flow cytometry. RESULTS: The median age was 49.0 years (range, 21~69) at transplantation. Univariate analysis showed that age less than 40 years old, lower frequencies of CD8+ T cells, invariant natural killer T (iNKT) cells, monocytic myeloid derived suppressor cells (M-MDSCs) and higher frequency of immature MDSCs were associated with occurrence of grade III–IV acute GVHD. Multivariate analyses showed that iNKT cells (hazard ratio (HR), 0.453, 95% CI, 0.091~0.844, p=0.024) and M-MDSCs (HR, 0.271, 95% CI, 0.078~0.937, p=0.039) were independent factors. Combination of higher frequencies of both cell subsets was associated with lower incidence of grade III–IV acute GVHD, whereas patients with lower frequency of iNKT cells and higher frequency of M-MDSCs showed significant higher probability of relapse. CONCLUSIONS: iNKT cells and M-MDSCs could be relevant cell biomarkers for predicting acute GVHD and/or relapse in acute leukemia patients treated with allo-HSCT.

Mucosal-associated invariant T cells and tumors / 中国药理学通报

Mucosal-assooiated invariant T(MAIT) cells are an evolutionarily highly conserved T lymphocyte subsets with the innate functions similar to innate natural killer T(iNKT) cells. MAIT cells are defined by their invariant T cell receptors (TCR)-alpha chain and restrictive major histocompatibility complex (MHC) related protein-! (MR1) , and identify antigens through the MR1, secreting a variety of cytokines after being activated, directly or indirectly involved in the body's immune re-sponses. MAIT cells are also abundant in human peripheral blood and many tissues. They are closely related to the occurrence and development of various infectious diseases, autoimmune diseases and malignant tumors. This article mainly reviews the research on MAIT cells in tumor diseases.

The clinical significance of invariant natural killer T cells in patients with human immunodeficiency virus infection / 中华传染病杂志

Objective@#To explore the changes of the peripheral invariant natural killer T (iNKT) cells in patients with human immunodeficiency virus (HIV) infection. @*Methods@#A total of 101 patients with HIV infection including 52 asymptomatic patients and 49 acquired immunodeficiency syndrome (AIDS) patients were enrolled in the study from June 2016 to July 2017. Flow cytometry was used to detect iNKT cells, CD4+ T cells and CD8+ T cells, and the relationship among them and HIV RNA was studied. At same time, 12 healthy persons were enrolled as control group. T test or variance analysis, rank sum test, Chi-square test and Fisher exact test were used for statistical analysis. @*Results@#In HIV infected asymptomatic patients, AIDS patients and healthy controls, iNKT cells were 0.135% (0.066%, 0.228%), 0.058% (0.034%, 0.100%) and 0.385% (0.205%, 0.600%), respectively, and the difference was statistical significant (Z=40.113, P<0.01). CD4+ T cell counts in the three groups were (340.82±119.26) cells/μL, (72.73±61.84) cells/μL and (555.17±229.43) cells/μL, respectively, and the difference was statistical significant (t=113.79, P<0.01); CD8+ T cell counts in the three groups were (842.29±423.68) cells/μL, (540.43±257.85) cells/μL and (875.92±516.45) cells/μL, respectively, and the difference was statistical significant (t=9.423, P<0.01). Ratios of CD4+ /CD8+ T cells in the three groups were 0.490 (0.240, 0.695), 0.120 (0.030, 0.210) and 0.600 (0.475, 0.895), respectively, and the difference was statistical significant (Z=53.603, P<0.01). iNKT cell counts in patients with or without hepatitis B virus infection, pneumocystis pneumonia, oral mold infection, treponema pallidum, latent tuberculosis or EB virus infection were not significantly different (Z=0.244, 2.325, 2.393, 0.168, 1.183 and 0.454, respectively, all P>0.05). There were correlations between iNKT cells and CD4+ T cells, CD4+ /CD8+ T cells (r=0.513 and 0.261, respectively, both P<0.05), and no relationship was found between iNKT cells and CD8+ T cells (r=0.155, P=0.126). In HIV infected asymptomatic patients and AIDS patients, iNKT cells was not associated with HIV RNA (r=-0.113 and -0.111, respectively, both P>0.05). @*Conclusions@#The level of peripheral iNKT cells in HIV infected patients decreases with the disease progression. To certain extent, iNKT cells can reflect the severity of immune damaging in HIV infected patients.

Deficiencies of Circulating Mucosal-associated Invariant T Cells and Natural Killer T Cells in Patients with Multiple Trauma

Mucosal-associated invariant T (MAIT) cells and natural killer T (NKT) cells are known to play important roles in autoimmunity, infectious diseases and cancers. However, little is known about the roles of these invariant T cells in multiple trauma. The purposes of this study were to examine MAIT and NKT cell levels in patients with multiple trauma and to investigate potential relationships between these cell levels and clinical parameters. The study cohort was composed of 14 patients with multiple trauma and 22 non-injured healthy controls (HCs). Circulating MAIT and NKT cell levels in the peripheral blood were measured by flow cytometry. The severity of injury was categorised according to the scoring systems, such as Acute Physiology and Chronic Health Evaluation (APACHE) II score, Simplified Acute Physiology Score (SAPS) II, and Injury Severity Score (ISS). Circulating MAIT and NKT cell numbers were significantly lower in multiple trauma patients than in HCs. Linear regression analysis showed that circulating MAIT cell numbers were significantly correlated with age, APACHE II, SAPS II, ISS category, hemoglobin, and platelet count. NKT cell numbers in the peripheral blood were found to be significantly correlated with APACHE II, SAPS II, and ISS category. This study shows numerical deficiencies of circulating MAIT cells and NKT cells in multiple trauma. In addition, these invariant T cell deficiencies were found to be associated with disease severity. These findings provide important information for predicting the prognosis of multiple trauma.

Clinical implications of circulating mucosal associated invariant T (MALT) cells in septic patients / 中国感染与化疗杂志

Objective To investigate the number and activation status of circulating mucosal associated invariant T (MAIT) cells in patients with sepsis.Methods Flow cytometric method was used to determine the number and percentage of MAIT cells and their expression of activation molecule CD69.Results The circulating MAIT cells dropped significantly at early stage of sepsis in septic patients.The MAIT cells in the sepsis group continued to express high levels of CD69 on day 1 and day 3 after admission to ICU.Conclusions Remarkable decrease of circulating MAIT cells may portend the possibility of sepsis in patients with severe infection.

Effects of invariant natural killer T cell activation on prognosis in early sepsis / 第二军医大学学报

Objective To explore the effects of invariant natural killer T (iNKT) cell activation on inflammation and severity of early sepsis. Methods Healthy C57BL/6 male mice were randomly divided into 3 groups: sham group, cecal ligation and puncture (CLP)-induced sepsis model group (CLP group) and CLP-induced sepsis model plus treatment with anti-CD1d blocking antibody group (anti-CD1d group). Flow cytometry was used to determine the frequencies of the iNKT cells in the peripheral blood, spleen and thymus of mice after 24 h of CLP operation. Meanwhile, the ELISA assay was used to detect the serum levels of tumor necrosis factor-α (TNF-α), interleukin (IL)-6, interferon-γ (IFN-γ) and IL-4. The correlation of iNKT cells and cytokine was analyzed. The bacterial colonies in peripheral blood and peritoneal lavage fluid were determined. The 10-day survival rates of mice after CLP operation were observed. Twenty patients with confirmed sepsis and 20 healthy volunteers aging 18-70 years were selected. The frequency of peripheral blood iNKT cells and serum cytokine levels of all patients and healthy volunteers were detected. Correlations of iNKT cells and cytokines, Acute Physiology and Chronic Health Evaluation Ⅱ (APACHE Ⅱ) scores in septic patients were analyzed. Results Compared with the sham group, the mice in the CLP group showed significantly more iNKT cells, significantly higher levels of TNF-α, IL-6, IFN-γ and IL-4 (P<0.05, P<0.01), and significantly higher levels of bacterial colonies in peripheral blood and peritoneal lavage fluid (P<0.01). The mice in the CLP group all died within 7 days after operation. In the mice of the anti-CD1d group, iNKT cell proportions in peripheral blood, spleen and thymus, the serum levels of TNF-α, IL-6, IFN-γ and IL-4 in peripheral blood, and the bacterial burden in the abdominal cavity were decreased compared with the CLP group (P<0.05, P<0.01). Four mice in anti-CD1d group survived at 10 days of post-operation. Compared with healthy volunteers, septic patients displayed significantly higher frequency of iNKT cells and significantly higher levels of TNF-α, IL-6, IFN-γ and IL-4 (P<0.05, P<0.01). Moreover, there was a positive correlation between frequency of iNKT cells with serum IFN-γ and IL-4 and the score of APACHE Ⅱ (P<0.05). Conclusion Increase of peripheral blood iNKT cells in early sepsis is associated with the severity and prognoses of sepsis, which suggests iNKT cells may play an important role in the inflammation of sepsis.

Research of functional segments of chicken B-FA molecule binding with Ii chain / 中国免疫学杂志

Objective: To research the functional segments of B-FA molecule binding invariant chain and their characters. Methods:The DNA segments (α1α2, sα1α2 and α3TC ) of B-FA genes were respectively cloned and inserted into prokaryotic or eukaryotic expression plasmids,then they were singly or co-transfected with Ii gene into the engineering bacteria E. coli (BL-21)or 293T cells. After induction of expression,affinity chromatography and SDS-PAGE identification,the binding between B-FA segments and Ii molecule and co-localization in cells were observed with Pull-down and Western blot. Results:First three recombinant prokaryotic expression plasmids and four recombinant eukaryotic expression plasmids were constructed. The single molecules expressed by B-FA segments were observed after an affinity chromatography. Secondly the complexes of Ii/B-FA-α1α2 and Ii/B-FA-sα1α2 were detected by a Pull-down from the co-transfected corresponding prokaryotic expression plasmids,but no complex of Ii andα3TC,also in the western blot it was detected that B-FA-α1α2 or B-FA-sα1α2 as functional segment could bind Ii to form complex. Finally in eukaryotic expression 293T cells B-FA-sα1α2 kept localization, the same as B-FA. Conclusion: Chicken B-FA-α1α2 is function segment to bind with Ii molecule and keeps the location characters same as B-FA. The results of this research first time provide experimental evidence about B-FA functional region binding segment to Ii molecule.

Effcets on immunoregulation of iNKT cells in RA by novel synthetic immunos-timulator CH1 b / 中国免疫学杂志

Objective:To investigate effects of a novel synthetic immunostimulator CH1b containing thiazolidin-4-one on the immunoregulation funotion of iNKT ( invariant nature killer T ) cells in active RA patients in vitro.Methods: Peripheral blood mononuclear cells( PBMCs) isolated from active RA patients were cultured with stimulation of α-Galcer and IL-2 in vitro and iNKT cells were then separated by using magnetic activated cell sorting( MACS) method with iNKT isolation kit.The cells were divided into three groups:control group (IL-2),α-Galcer group (IL-2+α-Galcer),CH1b group(IL-2 +CH1b).The effects of CH1b on the proliferation of iNKT cells in active RA patients were analyzed by using MTT assay.MILLIPLEX MAP Human Cytokine/Chemokine kit was used to evaluate the secretion of IFN-γand IL-4 in iNKT cells culture media.The expressions of IFN-γmRNA and IL-4 mRNA in iNKT cells were analyzed by RT-PCR.Results: Compared with control and α-Galcer group,the proliferation of iNKT cells of CH1b group were significantly higher( P<0.05).Compared with control,the ratio of IFN-γ/IL-4 in iNKT cells culture media in active RA patients of CH1b group were significantly lower (P<0.05).Compared with control,expressions of IFN-γmRNA and IL-4 mRNA were higher inα-Galcer group;compared with control,expressions of IL-4 mRNA were higher in CH1b group,while there were no obvious difference on expressions of IFN-γmRNA.Conclusion:CH1b was found to significantly stimulate the actived iNKT cells in active RA patients proliferation,promote the secretion of IL-4,and increase the ratio of IFN-γ/IL-4,promote the expression of IL-4 mRNA in iNKT cells in active patients.

Os princípios invariantes da noção de medida investigados a partir da perspectiva de sentido numérico / The invariant principles of measurement examined in the perspective of number sense / Los principios invariantes de la noción de medida investigados a partir de la perspectiva de sentido numérico

O presente estudo investiga o conhecimento de crianças sobre medidas a partir da noção de princípios invariantes e sentido numérico. Quarenta crianças (6 a 8 anos) foram solicitadas a fazer julgamentos acerca de situações envolvendo medidas de volume, massa, distância e comprimento. A Tarefa 1 examinou a habilidade de identificar unidades apropriadas para medir diferentes grandezas, e a Tarefa 2 examinou a compreensão acerca das relações inversas entre o tamanho da unidade e o número de unidades necessário para medir uma dada magnitude de um objeto. Os resultados mostraram que as crianças apresentam um sentido sobre medida o qual não se manifesta igualmente em relação a diferentes grandezas. Observou-se, também, que as crianças tinham uma melhor compreensão acerca das relações inversas entre o tamanho da unidade e o número de unidades do que acerca da relação entre unidade e a magnitude do objeto a ser medida.

This study investigates children's knowledge about measurements based on the idea of invariants principles, and of number sense. Forty children (6 and 8 years old) made judgments about situations involving measurements of volume, mass, distance and length. Task 1 examined the ability to identify the appropriate unit to measure different magnitudes, and Task 2 the understanding of the inverse relation between the size of the unit and the number of units needed to measure a magnitude of an object. Results showed that children have a sense of measurement and that this sense does not manifest itself equally in relation to different magnitudes. Also, children showed a better understanding about the inverse relation between the size of the unit and the number of units than of the relation between unit and the magnitude of the object to be measured.

El presente estudio investiga el conocimiento de los niños sobre medidas a partir de la noción de principios invariantes y sentido numérico. A cuarenta niños (6 a 8 años) les fue solicitado hacer juicios sobre situaciones que implicaban medidas de volumen, masa, distancia y longitud. La Tarea 1 examinó la habilidad para identificar unidades apropiadas para medir diferentes magnitudes y la Tarea 2 examinó la comprensión acerca de las relaciones inversas entre el tamaño de la unidad y el número de unidades necesario para medir una determinada magnitud de un objeto. Los resultados muestran que los niños presentan un sentido de medida que no se manifiesta de la misma forma en relación con diferentes magnitudes. También se observó que los niños tenían una mejor comprensión de las relaciones inversas entre el tamaño de la unidad y el número de unidades de que una comprensión de la relación entre unidades y la magnitud del objeto a ser medida.

Percentage and functions of natural killer T cells in peripheral blood and bone marrow of children with severe aplas-tic anemia / 临床儿科杂志

Objective To analyze the percentage and functional changes of natural killer T (NKT) cells in peripheral blood and bone marrow of severe aplastic anemia (SAA) children before immunosuppressive therapy (IST) comparing to that of healthy children.Methods Ten children with severe aplastic anemia were included in the study and ten healthy children at the same age were selected as the control group. By lfow cytometry, the percentage of CD3+CD1d tetramer+ NKT cell in peripheral blood and bone marrow were detected from March 2014 to December 2014 in our hospital. Immune magnetic bead separation was used to isolate and purify iNKT cells .The puriifed iNKT cells were cultured in the OCH(50 ng/ml,100 ng/ml or 200 ng/ml)+rhIL-2+rhG-CSF culture systems. The ampliifcation of iNKT cells after cultured in different systems were calculated. Elispot method was used to analyze the spotting form cells (SFCs) of IFN-γ or IL-4 expressed by activated iNKT cells.Results The percentage of CD3+CD1d tetramer+ NKT cells in peripheral blood of SAA group(0.72±0.03)% was signiifcantly lower than that of the control group(0.92±0.02)%(P=0.000). The percentage of CD3+CD1d tetramer+ NKT cells in bone marrow of SAA group(0.82±0.02)% was signiifcantly lower than that of the control group(1.05±0.05)%(P=0.000).In vitro iNKT cell ampliif-cation ability of bone marrow in SAA group was signiifcantly lower than the control group, and in medium concentration(50±6) and high concentration OCH group(52±6), the ampliifcation ability was higher than that in low concentration OCH group(30±5) (P<0.05). The secretion of IFN-γ in the iNKT cells of SAA bone marrow was signiifcantly lower in medium concentration(33±3) and high concentration(35±3)OCH group than that of the low concentration(50±3)OCH group(P<0.01). The secretion of IL-4 in the iNKT cells of SAA bone marrow was signiifcantly higher in medium concentration(50±3)and high concentration(75±3) OCH group than that of the low concentration(33±3) OCH group(P<0.01).Conclusions The quantity and function of NKT cells from children with SAA are lower than that of the healthy children.In vitro, they had better ampliifcation ability and could improve IL-4/IFN-γ imbalance in medium concentration and high concentration OCH group than in low concentration OCH group.

Mucosal-associated Invariant T cells: A New Player in Innate Immunity

Mucosal-associated invariant T (MAIT) cells are evolutionarily conserved T cells that are restricted by the non-classical major histocompatibility complex class-1b molecule MR1. MAIT cells recognize riboflavin (vitamin B2) derivatives in a MR1-dependent manner. Following antigen recongnition, MAIT cells rapidly produce Th1/Th17 cytokines, such as interferon-gamma and interleukin-17, in an innate-like manner. MAIT cells maintain an activated phenotype throughout the course of an infection, secrete inflammatory cytokines, and have the potential to directly kill infected cells, thus, playing an important role in controlling the host response. In this review, we discuss current knowledge regarding the role of MAIT cells in infectious diseases, cancers, and autoimmune diseases.

Deficiencies of Circulating Mucosal-associated Invariant T Cells and Natural Killer T Cells in Patients with Acute Cholecystitis

Mucosal-associated invariant T (MAIT) cells and natural killer T (NKT) cells are known to play crucial roles in a variety of diseases, including autoimmunity, infectious diseases, and cancers. However, little is known about the roles of these invariant T cells in acute cholecystitis. The purposes of this study were to examine the levels of MAIT cells and NKT cells in patients with acute cholecystitis and to investigate potential relationships between clinical parameters and these cell levels. Thirty patients with pathologically proven acute cholecystitis and 47 age- and sex-matched healthy controls were enrolled. Disease grades were classified according to the revised Tokyo guidelines (TG13) for the severity assessment for acute cholecystitis. Levels of MAIT and NKT cells in peripheral blood were measured by flow cytometry. Circulating MAIT and NKT cell numbers were significantly lower in acute cholecystitis patients than in healthy controls, and these deficiencies in MAIT cells and NKT cell numbers were associated with aging in acute cholecystitis patients. Notably, a reduction in NKT cell numbers was found to be associated with severe TG13 grade, death, and high blood urea nitrogen levels. The study shows numerical deficiencies of circulating MAIT and NKT cells and age-related decline of these invariant T cells. In addition, NKT cell deficiency was associated with acute cholecystitis severity and outcome. These findings provide an information regarding the monitoring of these changes in circulating MAIT and NKT cell numbers during the course of acute cholecystitis and predicting prognosis.

Effects of a novel synthetic immunostimulator CH2b on iNKT cells isolated from patients with active rheumatoid arthritis / 中华微生物学和免疫学杂志

Objective To investigate the effects of a novel synthetic immunostimulator CH2b containing thiazolidin-4-one on the function of invariant nature killer T (iNKT) cells isolated from patients with active rheumatoid arthritis (RA).Methods Peripheral blood mononuclear cells (PBMCs) isolated from patients with active RA were in vitro cultured with α-Galcer and IL-2.The iNKT cells were separated by using magnetic activated cell sorting (MACS) method.The effects of CH2b on the proliferation of iNKT cells were analyzed by using MTT assay.MILLIPLEX MAP Human Cytokine/Chemokine kit was used to measure the levels of IFN-γ and IL-4 in the supernatants of iNKT cell culture.The expressions of IFN-γand IL-4 at mRNA level in iNKT cells were analyzed by RT-PCR.Western blot assay was used to detect the levels of T-bet and GATA-3 in iNKT cells.Results CH2b significantly enhanced the proliferation of IL-2 activated iNKT cells isolated from the patients with active RA.CH2b promoted the secretion of IL-4,resulting in a decrease in the ratio of IFN-γ/IL-4.Moreover,CH2b promoted the expressions of GATA-3 and IL-4 at mRNA level in iNKT cells.Conclusion The novel immunostimulator,CH2b,might enhance the immunoregulatory effects of iNKT cells by promoting the GATA-3 pathway-mediated secretion of Th2-1ike cytokines and inducing the differentiation of Th0 to Th2 cells.

Characters of location in cells and binding activity of chicken Ii and B-L gene ex-pressed products / 中国免疫学杂志

Objective:To study characters of the location in cells and binding activity of chicken Ii and B-L gene expressed products.Methods:The cloned gene segments of chicken Ii,B-LA and B-LB were respectively inserted into prokaryotic or eukaryotic expression plasmids,and then these recombinant plasmids were respectively alone transfected or cotransfected into engineering bacteria, Rosetta(DE3) or 293T cells.All of the recombinant bacteria were induced to express and their products then were renatured.The singly expressed products were detected to their immunogenicity with Western blot, and the co-expressed products were tested their binding with pull-down method and ( SDS-) PAGE.Results:First six of prokaryotic and eukaryotic recombinant expression plasmids were con-structed.The eukaryotic expressed products of Ii, B-LA and B-LB genes located in cellular plasma membrane.The single protein molecules were achieved from prokaryotic expressed products, which were renatured and purified with a Ni-column respectively.Secondly the prokaryotic expression Ii,B-LA and B-LB molecules could respectively induce mouse to product specific anti-bodies, which could recognize the corresponding products in eukaryotic expression.This indicated that they retained their antigenicity.Finally with pull-down from the products in co-expression in engineering bacteria and renaturation the Ii/B-LA or Ii/B-LB complexes were purified and the Ii,B-LA or B-LB monomers were dissociated from these complexes after a SDS treatment.Conclusion:The prokaryotic or eukaryotic expressed products of chicken Ii and B-L genes could retain their antigenicity,and chicken Ii could bind B-L molecules after prokaryotic coexpression and renaturation.These results provide a useful method to study the relation between Ii and MHC molecules.