Metformin Inhibits ROS/TNF-alpha Axis-Mediated Chronic Kidney Disease Induced by TAA Independent of Leukocyte Infiltration in Association with the Inhibition of Kidney Injury Biomarkers / La Metformina Inhibe la Enfermedad Renal Crónica Mediada por el Eje ROS/TNF-alfa Inducida por TAA Independientemente de la Infiltración de Leucocitos en Asociación con la Inhibición de Biomarcadores de Lesión Renal

SUMMARY: The toxic effects of thioacetamide (TAA) and carbon tetrachloride on the human body are well recognized. In this study, we examined whether TAA intoxication can induce kidney leukocyte infiltration (measured as leukocyte common antigen CD45) associated with the augmentation of the reactive oxygen species (ROS)/tumor necrosis factor-alpha (TNF-α) axis, as well as biomarkers of kidney injury with and without metformin treatment. Rats were either injected with TAA (200 mg/kg; twice a week for 8 weeks) before being sacrificed after 10 weeks (experimental group) or were pre-treated with metformin (200 mg/kg) daily for two weeks prior to TAA injections and continued receiving both agents until the end of the experiment, at week 10 (protective group). Using basic histology staining, immunohistochemistry methods, and blood chemistry analysis, we observed profound kidney tissue injury such as glomerular and tubular damage in the experimental group, which were substantially ameliorated by metformin. Metformin also significantly (p0.05) increase in kidney expression of CD45 positive immunostaining cells. In conclusion, we found that TAA induces kidney injury in association with the augmentation of ROS/TNF-α axis, independent of leukocyte infiltration, which is protected by metformin.

Son bien conocidosos los efectos tóxicos de la tioacetamida (TAA) y el tetracloruro de carbono en el cuerpo humano. En este estudio, examinamos si la intoxicación por TAA puede inducir la infiltración de leucocitos renales (medida como antígeno leucocitario común CD45) asociada con el aumento de las especies reactivas de oxígeno (ROS)/factor de necrosis tumoral-alfa (TNF-α), así como biomarcadores de daño renal con y sin tratamiento con metformina. A las ratas se les inyectó TAA (200 mg/kg; dos veces por semana durante 8 semanas) antes de sacrificarlas a las 10 semanas (grupo experimental) o se les pretrató con metformina (200 mg/kg) diariamente durante dos semanas antes de las inyecciones de TAA y continuaron recibiendo ambos agentes hasta el final del experimento, en la semana 10 (grupo protector). Usando tinción histológica básica, métodos de inmunohistoquímica y análisis químico de la sangre, observamos una lesión profunda del tejido renal, como daño glomerular y tubular en el grupo experimental, que mejoraron sustancialmente con la metformina. La metformina también inhibió significativamente (p0,05) en la expresión renal de células de inmunotinción positivas para CD45. En conclusión, encontramos que el TAA induce la lesión renal en asociación con el aumento del eje ROS/TNF-α, independientemente de la infiltración de leucocitos, que está protegida por metformina.

Suppression of diabetes-induced renal inflammatory cell infiltration by metformin associated with the amelioration of renal injury biomarkers / Supresión de la infiltración de células inflamatorias renales de diabetes-inducida, por metformina asociada con la mejora de los biomarcadores de lesión renal

SUMMARY: Diabetes and hypertension account for the majority of chronic kidney injury cases that can lead to renal failure. The link between the leukocytes common antigen (CD45) and diabetic kidney disease (DKD) with and without metformin incorporation in an animal model has not been investigated before. Therefore, we sought to assess the extent of leukocytes infiltration into kidney tissues 10 weeks following the induction of diabetes in rats treated with metformin. In addition, we monitored blood and urine parameters associated with diabetes. The model group of rats received streptozotocin (STZ; 50 mg/kg) injection after being fed for 14 days on a high-fat diet (HFD) and continuously fed a HFD until they were culled, at week 12. The protective group was treated in the same way except that these animals were put from day 1 on metformin (200 mg/kg) until being culled, on week 12. Kidneys were immunostained with CD45 as a marker of leukocytes infiltration and examined by light microscopy. Urine samples were tested for urine albumin and collected blood was analyzed for sugar, urea, creatinine, and oxidative stress and antioxidants biomarkers. Kidney injury secondary to diabetes was developed as demonstrated by (i) increased blood glucose, urea, and malondialdehyde (MDA) as a marker of lipid peroxidation; and (ii) kidney tissue damage and marked increase in kidney tissues expressing CD45 positive cells. The above markers were inhibited (p0.0006) by metformin. Also, a significant correlation was observed between CD45 score and glycemia, urea, MDA, and the antioxidant superoxide dismutase (SOD). Thus, our data demonstrate an association between the infiltration of CD45+ inflammatory cells into kidney tissues and biomarkers of kidney damage in a rat model of DKD, which was effectively protected by metformin.

RESUMEN: La diabetes y la hipertensión representan la mayoría de los casos de lesión renal crónica que pueden provocar insuficiencia renal. El vínculo entre el antígeno común de los leucocitos (CD45) y la enfermedad renal diabética (DKD) con y sin incorporación de metformina en un modelo animal no se había anteriormente investigado. El objetivo fue evaluar el grado de infiltración de leucocitos en los tejidos renales 10 semanas después de la inducción de diabetes en ratas tratadas con metformina. Además, monitoreamos los parámetros de sangre y orina asociados con la diabetes. El grupo modelo de ratas recibió una inyección de estreptozotocina (STZ; 50 mg/kg) después de ser alimentadas durante 14 días con una dieta alta en grasas (HFD) y continuamente alimentadas con un HFD hasta que fueron sacrificadas, en la semana 12. El grupo protector fue tratado de la misma manera excepto que estos animales fueron recibieron desde el día 1 metformina (200 mg/kg) hasta ser sacrificados, en la semana 12. Los riñones fueron inmunoteñidos con CD45 como marcador de infiltración de leucocitos y examinados por microscopía óptica. Las muestras de orina se analizaron en busca de albúmina y la sangre recolectada se analizó en busca de glucosa, urea, creatinina y biomarcadores de estrés oxidativo y antioxidantes. La lesión renal secundaria a la diabetes se desarrolló como lo demuestra (i) el aumento de la glucosa en sangre, la urea y el malondialdehído (MDA) como marcador de la peroxidación lipídica; y (ii) daño del tejido renal y marcado aumento en los tejidos renales que expresan células positivas para CD45. Los marcadores anteriores fueron inhibidos (p≤0.0006) por metformina. Además, se observó una correlación significativa entre la puntuación de CD45 y la glucemia, la urea, la MDA y la superóxido dismutasa antioxidante (SOD). Por lo tanto, nuestros datos demuestran una asociación entre la infiltración de células inflamatorias CD45+ en los tejidos renales y biomarcadores de daño renal en un modelo de rata con DKD, que fue protegido de manera efectiva por metformina.

Paracetamol poisoning induces acute liver injury in rats: inhibition of miR-155/cd45 axis-mediated antioxidant depletion and hepatotoxicity using quercetin and resveratrol / La intoxicación por paracetamol induce daño hepático agudo en ratas: inhibición de la depleción de antioxidantes mediado por el eje miR-155/cd45 y hepatotoxicidad usando quercetina y resveratrol

SUMMARY: Ingestion of an overdose of paracetamol (also called acetaminophen, or APAP) induces hepatotoxicity that can lead to liver failure. The link between the pro-inflammatory microRNA-155 (miR-155) and leukocyte infiltration (CD45) in APAP- antioxidant depletion and liver toxicity with and without the natural polyphenolic compounds, quercetin (QUR) plus resveratrol (RES) has not been previously studied. Therefore, acute hepatic injury was induced in rats by 2 g/kg APAP (single dose, orally) and another group started QUR (50 mg/kg) plus RES (30 mg/kg) treatment one week prior to APAP ingestion. Animals were culled 24 hours post the paracetamol treatment. APAP overdose induced hepatic and blood levels of miR-155 expression, CD45 (leukocyte common antigen) immunostaining, degenerated hepatocytes, and hepatic injury enzymes; alanine aminotransferase (ALT) and aspartate aminotransferase (AST), which were markedly decreased by QUR+RES. Whereas, APAP intoxication ameliorated liver tissue levels of the antioxidants, glutathione peroxidase and superoxide dismutase that were augmented by QUR+RES. Moreover, a significant (p<0.05) correlation between miR-155/CD45 axis and liver tissue injury was observed. These findings show that paracetamol intoxication augments miR- 155/CD45 axis-mediated modulation of antioxidants and liver injury in rats, and is protected by QUR+RES.

RESUMEN: La ingestión de una sobredosis de paracetamol (también llamado acetaminofeno o APAP) induce hepatotoxicidad que puede provocar insuficiencia hepática. El vínculo entre el microARN-155 proinflamatorio (miR-155) y la infiltración de leucocitos (CD45) en el agotamiento de APAP- antioxidante y la toxicidad hepática con y sin los compuestos polifenólicos naturales, quercetina (QUR) más resveratrol (RES) no ha sido previamente investigado. En este estudio, se indujo daño hepático agudo en ratas con 2 g/kg de APAP (dosis única, por vía oral) y otro grupo comenzó el tratamiento con QUR (50 mg/ kg) más RES (30 mg/kg) una semana antes de la ingestión de APAP. Los animales se sacrificaron 24 horas después del tratamiento con paracetamol. La sobredosis de APAP indujo niveles hepáticos y sanguíneos de expresión de miR-155, inmunotinción de CD45 (antígeno leucocitario común), degeneración de los hepatocitos y daño hepático enzimático; alanina aminotransferasa (ALT) y aspartato aminotransferasa (AST), disminuyeron notablemente con QUR+RES. Mientras que la intoxicación con APAP mejoró los niveles de antioxidantes, glutatión peroxidasa y superóxido dismutasa en el tejido hepático los que aumentaron con QUR+RES. Además, se observó una correlación significativa (p<0,05) entre el eje miR-155/CD45 y la lesión del tejido hepático. Estos hallazgos muestran que la intoxicación por paracetamol aumenta la modulación mediada por el eje miR-155/CD45 de los antioxidantes y la lesión hepática en ratas, y está protegida por QUR+RES.

CD45 y leucemia linfoide aguda pediátrica / CD45 and childhood acute lymphoblastic leukemia

Introducción: El CD45 se expresa en las células hematopoyéticas, su determinación es indispensable para la clasificación inmunofenotípica de la leucemia linfoide aguda (LLA). Objetivo: Evaluar la expresión del antígeno CD45 en los blastos de pacientes pediátricos con LLA y su relación con las características biológicas, morfológicas y clínicas al inicio de la enfermedad, la respuesta al tratamiento y la supervivencia global (SG) de los enfermos. Métodos: Se estudiaron 160 pacientes con LLA entre diciembre del 2012 y diciembre del 2017, tratados con el protocolo ALL-IC BFM-SG 2009. El inmunofenotipaje celular de la médula ósea se realizó por citometría de flujo. Resultados: El fenotipo B CD45+ predominó en los menores de seis años de edad y en los mayores de diez, el fenotipo T CD45+. Se encontró diferencia significativa entre la ausencia de adenopatías mediastínicas, el fenotipo leucémico y la ausencia de CD45 (p=0.004); entre la respuesta a la prednisona en sangre periférica al día ocho, el fenotipo leucémico y la ausencia de CD45 (p=0.001). Se encontraron diferencias significativas entre la respuesta a la prednisona en sangre periférica el día ocho y la respuesta en médula ósea el día 33, según fenotipo leucémico (p=0.009) y la presencia en los blastos del antígeno CD45 (p=0.02). Se encontró diferencia significativa entre la SG de los enfermos, según fenotipo leucémico y la ausencia del antígeno CD45 (p=0.017). Conclusión: La expresión o ausencia del antígeno de CD45 en los blastos tiene relación con la respuesta al tratamiento y la SG de pacientes pediátricos con LLA(AU)

Introduction: CD45 is expressed in hematopoietic cells, its determination is essential for the immunophenotypic classification of acute lymphoid leukemia (ALL). Objective: To evaluate the expression of the CD45 antigen in the blasts of pediatric patients with ALL and its relationship with the biological, morphological and clinical characteristics at the onset of the disease, the response to treatment and the overall survival (OS) of the patients. Methods: 160 patients with ALL were studied between December 2012 and December 2017, treated with the ALL-IC BFM-SG 2009 protocol. Bone marrow cellular immunophenotyping was performed by flow cytometry. Results: Patients with the CD45 + B phenotype predominated in those under six years of age, while those with a CD45 + T phenotype in those older than ten. A significant difference was found between the absence of mediastinal lymph nodes, the leukemic phenotype and the absence of CD45 (p = 0.004). A significant difference was found between the response to prednisone in peripheral blood at day eight, the leukemic phenotype and the absence of CD45, p = 0.001. Significant differences were found between the response to prednisone in peripheral blood on day eight and the response in bone marrow on day 33, according to leukemic phenotype and the presence in blasts of the CD45 antigen (p = 0.009 and p = 0.02, respectively). A significant difference was found between the OS of patients, according to leukemic phenotype and the absence of the CD45 antigen, p = 0.017. Conclusion: The expression or absence of the CD45 antigen in blasts is related to the response to treatment and OS of pediatric patients with ALL(AU)

Subsets of T lymphocytes in the lesional skin of pityriasis rosea

Abstract: Background: Pityriasis rosea is a common papulosquamous disorder. However, its etiology and pathogenesis remain unclear. Objective: We investigate the types of inflammatory cells infiltrating the lesional skin of pityriasis rosea and demonstrate whether T-cell-mediated immunity is involved in the pathogenesis of this condition or not. Methods: The biopsies were taken from the lesional skin of 35 cases of patients diagnosed with pityriasis rosea. The specimens were prepared in paraffin sections, then submitted to routine immunohistochemistry procedures using monoclonal antibodies directed against CD3, CD4, CD8, CD20 and CD45RO and horseradish peroxidase-labeled goat anti-human antibodies. The positive sections were determined by the ratio and staining intensity of positive inflammatory cells. Results: The mean score of positive CD3, CD4, CD8, and CD45RO staining was respectively 3.74±3.88, 5.67±4.40, 2.94±3.42 and 7.68±4.33 in these pityriasis rosea patients (P<0.001). The percentage of positive staining was 54.29% (19/35), 69.7% (23/33), 40% (14/35) and 79.41% (27/34) (P<0.05). However, the staining of CD20 was negative in all samples. The mean score of CD3 staining in patients with time for remission ≤60 days (4.90±4.21) was higher than that in patients with time for remission >60 days (2.00±2.5) (P<0.05), whereas no statistical difference in the mean score of CD4, CD8 and CD45RO staining was observed. study liMitations: The sample size and the selected monoclonal antibody are limited, so the results reflect only part of the cellular immunity in the pathogenesis of pityriasis rosea. Conclusion: Our findings support a predominantly T-cell mediated immunity in the development of pityriasis rosea.

Prognostic Significance of CD45CD117 Cells in Patients with Acute Myeloid Leukemia after Complete Remission / 中国实验血液学杂志

OBJECTIVE@#To investigate the predictive value of CD45CD117 phenotype-abnormal cells (hereinafter referred to as "abnormal cells") for relapse and prognosis in adult patients with acute myeloid leukemia (AML) within 2 weeks after the first complete remission (CR1).@*METHODS@#The clinical data of patients with newly diagnosed AML (non-acute promyelocytic leukemia) admitted in our department from July 1, 2014 to June 30, 2017 were analyzed retrospectively, and the relationship between clinical features at the initial diagnosis and the abnormal phenotype cells of CD45CD117 within 2 weeks after CR1 with the prognosis were analyzed.@*RESULTS@#A total of 91 patients with CD45CD117 abnormal cells were detected. The median age was 51 years old, the median WBC count was 11.60×109/L, and the median ratio of bone marrow blast cells was 0.35 at initial diagnosis. According to the FAB classification, 1 (1.1%), 7 (7.7%), 38 (41.7%), 20 (22.0%), 21 (23.1%) and 4 (4.4%) patients were classifice as M0, M1, M2, M4, M5, and M6, respectively. According to the NCCN risk stratification, 30 (33.0%), 51 (56.0%), and 10 (11.0%) patients were determined as good, moderate, and poor prognosis, respectively. The median ratio of abnormal cells within 2 weeks after CR1 was 1.8500 (0.0236-8.0000)%. The median time from initiation of induction therapy to the acquisition of CR was 46 days, median recurrence-free survival time was 319 days, and median overall survival time was 352 days. A total of 45 patients relapsed, of which 14 died; 46 patients did not relapse, of which 3 died. The cutoff of abnormal cells by receiver operating characteristic curve (ROC) analysis was 2.055% (Se=0.733，Sp=0.761). The abnormal cell ratio was＞2.055% in 44 patients, the median ratio of abnormal cells was 3.075%, among which 33 patients relapsed and 12 patients died; the abnormal cell ratio was ＜2.055% in 47 patients, the median ratio of abnormal cells was 1.150%, 12 patients relapsed and 5 patients died. Regression analysis showed that WBC count＞50×10/L and abnormal cell ratio＞2.055% were independent risk factors for recurrence. The abnormal cell ratio＞2.055% group had a 2-year RFS rate of 54.3% and a 2-year OS rate of 52.8%. The abnormal cell ratio＜2.055% group had a 2-year RFS rate of 86.6% (P=0.018), and a 2-year OS rate of 85.3% (P＜0.05).@*CONCLUSION@#For adult AML patients, CD45CD117 phenotypical abnormal cells ratio＞2.055% within 2 weeks after CR1 is an independent risk factor for recurrence, which also is an dverse factor for RFS and OS.

Antígenos MICA y trasplante / MICA antigens and transplantation

Los antígenos leucocitarios humanos (HLA, del inglés human leukocyte antigens), codificados por los genes del complejo principal de histocompatibilidad (MHC, del inglés m ajor histocompatibility complex), actúan como inductores de las respuestas inmunitarias en el trasplante; sin embargo, los productos de los genes relacionados a cadenas MHC clase I (MIC, del inglés MHC class I chain-related genes), constituyen también uno de los blancos del rechazo. La familia de los genes MIC consta de siete miembros, de los cuales solo MICA y MICB son funcionales. Los transcriptos son glicoproteínas de superficie celular de 62 kDA que presentan homología en su secuencia con las moléculas HLA clase I y cuya función está relacionada con la inmunidad innata. En los órganos trasplantados ocurre un incremento en la expresión de los antígenos MICA como una señal temprana de "peligro" debido al trauma quirúrgico y la isquemia. Esta sobrexpresión antigénica puede llevar al rechazo mediado por anticuerpos anti-MICA que activan el complemento y por un incremento de la citotoxicidad debido a la estimulación en los linfocitos citolíticos naturales (NK, del inglés natural killer) y los linfocitos CD8+ &+ αß y γδ, del receptor conocido como NKG2D (NK grupo 2 miembro D(AU)

Human leukocyte antigens (HLA), encoded by major histocompatibility complex (MHC) genes, act as inducers of immune responses in transplantation. However, the products of the genes related to MHC class I chains (MIC) are also one of the targets of rejection. The family of MIC genes consists of seven members, of which only MICA and MICB are functional. Transcripts are cell surface glycoproteins of 62 kDa that exhibit homology in sequence with HLA class I molecules and whose function is related to innate immunity. In transplanted organs an increase in the expression of MICA antigens occurs as an early sign of "danger" due to surgical trauma and ischemia. This antigenic overexpression can lead to rejection mediated by complement-activating anti-MICA antibodies and by increased cytotoxicity due to stimulation in natural killer (NK) lymphocytes and CD8 + + αß and γδ lymphocytes. Receptor known as NKG2D (NK group 2 member D)(AU)

Expresión del antígeno CD45 en la Leucemia Linfoide Aguda Pediátrica / Evaluation of CD45 antigen expression in acute lymphoblastic leukemia

Introducción: la leucemia linfoide aguda (LLA) es la neoplasia más frecuente en la infancia. La determinación del antígeno CD45 discrimina entre los blastos y las células reactivas en la médula ósea (MO). Objetivo: evaluar la expresión del antígeno CD45 sobre los blastos de pacientes con LLA, según los distintos subtipos inmunológicos, su posible relación con las características biológicas y clínicas de presentación de la enfermedad y la respuesta al tratamiento antileucémico. Métodos: se estudiaron 150 pacientes con LLA procedentes de varios servicios oncohematológicos del país, entre enero del 2008 y mayo del 2015. El inmunofenotipaje celular de la MO se realizó por citometría de flujo. Resultados: el antígeno CD45 mostró una gran heterogeneidad de expresión sobre los linfoblastos. Del total de enfermos estudiados, 19,3 por ciento no expresaron sobre los blastos el antígeno CD45, 36,7 por ciento presentaron una expresión moderada y 44 por ciento mostraron una alta densidad de expresión. Se encontró diferencia significativa al comparar el fenotipo leucémico con la expresión del antígeno CD45 sobre los blastos (p = 0,000). Ningún enfermo presentó adenopatías mediastinales, con diferencias significativas (p = 0,000), según el fenotipo y la expresión de CD45. Los pacientes con LLA-T cuyos blastos no expresaron CD45 tuvieron una mala respuesta al tratamiento anti-leucémico los días 8 y 15 en sangre periférica y MO, respectivamente. Conclusión: la expresión de CD45 sobre los blastos, pudiera ser considerada como un factor pronóstico adicional para la estratificación en diferentes grupos de riesgos, de la LLA en el niño(AU)

Introduction: Acute lymphoblastic leukemia (ALL) is the most frequent neoplasia in infancy. Determination of CD45 antigen discriminates between blasts and reactive cells in the bone marrow (MO). Objective: To evaluate the expression of the CD45 antigen on the blasts of patients with ALL, according to the different immunological subtypes, their possible relation with the biological and clinical characteristics of the disease and the response to antileukemic treatment. Methods : 150 patients with ALL were studied from various onco-hematological services of the country, between January 2008 and May 2015. The cellular immunophenotyping of the MO was performed by flow cytometry. Results : The CD45 antigen showed a great heterogeneity of expression on the lymphoblasts. Of the total number of patients studied, 19.3 percent did not express the CD45 antigen on the blasts, 36.7 percent presented moderate expression and 44 percent showed a high expression density of it.A significant difference was found when comparing the leukemic phenotype with the expression of the CD45 antigen on the blasts (p = 0.000). No patient had mediastinal lymphadenopathy, with significant differences (p = 0.000), according to the phenotype and CD45 expression. Patients with T-ALL whose blasts did not express CD45 had a poor response to anti-leukemic treatment on days 8 and 15 in peripheral blood and MO, respectively. Conclusion: CD45 expression on blasts could be considered as an additional prognostic factor for stratification in different risk groups of ALL in children(AU)

Immunophenotypic Analysis of Acute Promyelocytic Leukemia / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the immunophenotype of leukemia promyelocytes (LP) in bone marrow of patients with acute promyelocytic leukemia (APL) and to explore their characteristics and significance.</p><p><b>METHODS</b>The immunophenotypes of leukemia cells in 43 patients with APL were analyzed by means of 4 color immunophenotypes; the cell population in which CD45 strength localized at 10(2) and the SSC strength locatized at 10(2) was defined as R3, the cell population in which CD45 strength localized at 10(3) and the SSC strength localized at 10(2) was defined as R5, moreover the ratio of positive cells >80% was defined as strong positive expression, the ratio of positive cells between 20%-80% was difined as weak positive expression, the ratio of positive cells <20% was difined as negative by gating method of CD45/SSC.</p><p><b>RESULTS</b>There was a abnormal cell population (R3) in 79.07% cases; the immunophenotypes of R3 was cheracteried by high SSC, weaker expression of CD45, the rate of CD38, CD9 and CD13 all was 100%, moreover their bright expression (>80%) was 86.05%, 90.70% and 86.05%, respectively; the positive expression rate of CD33, CD117 and CD64 was 97.67%, 95.35% and 83.80% respectively, moreover thier bright expression was 84.04%, 69.77% and 30.23% respectively; the CD15 was weakly expressed in 39.53% cases, the CD34 and HLA-DR were weakly expression in 16.28% and 6.98% cases respectively. All the cases did not express CD116. There were 2 cell populations (R3 and R5) in 20.93% cases, the immunophenotypic features of R3 were cosistant with above mentioning, while the immunophenotypes of R5 were lower than those of R3 SSC; the fluorescence intensity of CD45 was higher, but lower than that in normal lymphycytes, the positive rate of CD9, CD13, MPO was 100%, moreover thier fluorescence intensity was high; they did not expressed CD123, CD25, CD22, CD4, CD64 and CD14. Thereby it can be concluded that the typical immunophenotypes is characterized by CD13(+) CD9(+) CD38(+) CD33(+) CD117(+) CD64(+) CD11b(-) CD34(-) HLA-DR(-) in APL. There was a special immunophenotype in the APL with basophilic granules. Conclusoin: APL has a characteristic immunophenotypic profile, whose typical immunophenotype is characterized by CD13(+) CD9(+) CD38(+) CD33(+) CD117(+) CD64(+) CD11b(-) CD34(-) HLA-DR(-). The special immunophenotype exists in the APL with basophilic granules. Flow cytometric immunophenotyping may be a useful for rapid recognition of APL and has significant for prognosis.</p>

Transformation of patient-derived tumor xenografts into lymphomas: characteristics, influence factors and precautions / 中华胃肠外科杂志

The patient-derived tumor xenografts (PDX) model is an animal model established by directly engrafting fresh tumor tissue of patients into immunodeficiency mice after surgery or biopsy, which plays an important role in the study of tumor biology. However, the transformation of PDX into lymphoma limits the application of this model. The characters of this transformation include that epithelial tumors origin, predorminance of B-cell lymphomas, lost of architectural feature of primary tumor, absence of epithelial tumor markers, and CD45 and CD20 expression. That were characteristics of human B lymphocytes, and possible infection of Epstein-Barr virus(EBV). The biology of primary tumor, EBV infection, inflammation infiltration in primary tumors and the host immune status are the main related factors in this transformation. Therefore, selective xenograft by the detection of EBV infection and inflammation infiltration in primary tumors may be effective methods to prevent lymphomagenesis.

Intravenous transplantation of very small embryonic like stem cells in treatment of diabetes mellitus

Diabetes Mellitus [DM], simply known as diabetes, refers to a group of metabolic diseases in which there are high blood sugar levels over a prolonged period. In this study, the feasibility and safety of intravenous transplantation of Very Small Embryonic Like stem cells [VSELs] were investigated for diabetes repair, and finally the migration and distribution of these cells in hosts were observed. Mouse bone marrow VSELs were isolated by Fluorescent Activating Cell Sorting [FACS] method by using fluorescent antibodies against CD45, CXCR4 and Sca1 markers. Sorted cells were analyzed for expression of oct4 and SSEA1 markers with immunocytochemistry staining method. To determine multilineage differentiation, sorted cells were differentiated to Schwann, osteocyte and beta cells. Ten days after the establishment of a mouse model of pancreas necrosis, Dil-labeled VSELs were injected into these mice via tail vein. Pancreases were harvested 4 weeks after transplantation and the sections of these tissues were observed under fluorescent microscope. It was proved that CD45-, CXCR4+, and Sca1+ sorted cells express oct4 and SSEA1. Our results revealed that intravenously implanted VSELs could migrate into the pancreas of hosts and survive in the diabetic pancreas. In treated groups, blood glucose decreased significantly for at least two month and the weights of mice increased gradually. This study provides a strategy for using VSELs for curing diabetes and other regenerative diseases, and the strategy is considered an alternative for other stem cell types

Cloning of human CD45 gene and its expression in Hela cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To clone human CD45 gene PTPRC and establish Hela cells overexpressing recombinant human CD45 protein.</p><p><b>METHODS</b>The intact cDNA encoding human CD45 amplified using RT-PCR from the total RNA extracted from peripheral blood mononuclear cells (PBMCs) of a healthy donor was cloned into pMD-18T vector. The CD45 cDNA fragment amplified from the pMD-18T-CD45 by PCR was inserted to the coding region of the PcDNA3.1-3xflag vector, and the resultant recombinant expression vector PcDNA3.1-3xflag-CD45 was transfected into Hela cells. The expression of CD45 in Hela cells was detected by flow cytometry and Western blotting, and the phosphastase activity of CD45 was quantified using an alkaline phosphatase assay kit.</p><p><b>RESULTS</b>The cDNA fragment of about 3 900 bp was amplified from human PBMCs and cloned into pMD-18T vector. The recombinant expression vector PcDNA3.1-3xflag-CD45 was constructed, whose restriction maps and sequence were consistent with those expected. The expression of CD45 in transfected Hela cells was detected by flow cytometry and Western blotting, and the expressed recombinant CD45 protein in Hela cells showed a phosphastase activity.</p><p><b>CONCLUSION</b>The cDNA of human CD45 was successfully cloned and effectively expressed in Hela cells, which provides a basis for further exploration of the functions of CD45.</p>

Gastric Perforation Caused by Primary Gastric Diffuse Large B Cell Lymphoma / 대한소화기학회지

Spontaneous gastric perforation is a rare complication of gastric lymphoma that is potentially life threatening since it can progress to sepsis and multi-organ failure. Morbidity also increases due to prolonged hospitalization and delay in initiating chemotherapy. Therefore prompt diagnosis and appropriate treatment is critical to improve prognosis. A 64-year-old man presented to the emergency department with severe abdominal pain. Chest X-ray showed free air below the right diaphragm. Abdominal CT scan also demonstrated free air in the peritoneal cavity with large wall defect in the lesser curvature of gastric lower body. Therefore, the patient underwent emergency operation and primary closure was done. Pathologic specimen obtained during surgery was compatible to diffuse large B cell lymphoma. Fifteen days after primary closure, the patient received subtotal gastrectomy and chemotherapy was initiated after recovery. Patient is currently being followed-up at outpatient department without any particular complications. Herein, we report a rare case of gastric lymphoma that initially presented as peritonitis because of spontaneous gastric perforation.

Effect of Shen warming Pi strengthening method on the expression of serum T cell subsets in IBS-D rats / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To observe the effect of Shen warming Pi strengthening method on expressions of serum T cell subsets (C045+%, C03+%, and C04 +/COB+) in diarrhea-predominant irritable bowel syndrome (IBS-0) rats. Methods An IBS-0 rat model was established referring to AL-Chaer's modeling method combined with tail clamp and intragastric administration of sanna leaf. After modeling 30 SO rats were randomly divided into 6 groups according to random digit table, i.e., the model group, the high, middle, low dose Wenshen Jianpi Recipe (WJR) groups, and the Sishen Pill control group, 6 in each group. A normal control group consisting of 6 SO rats were also set up. Rats in high, middle, low dose WJ R groups were administered by gastrogavage with boil-free WJ R at the daily dose of 3. 100, 1. 550, 0. 775 g/kg, respectively. Rats in the Sis hen Pill control group were administered by gastrogavage with boil-free Sis hen Pill at the daily dose of 0. 736 g/kg. Equal volume of normal saline was given by gastrogavage to rats in the model group and the normal control group. All medication lasted for 2 successive weeks. Rats' general state, expressions of T cell subsets (CD45+%, CD3+%, and CD4+ /CDB+) changes were observed.</p><p><b>RESULTS</b>Compared with the normal control group, expressions of CD45+% and CD3+% increased, but CD4+ /CDB+ decreased with statistical difference (P < 0. 05). Compared with the model group, expressions of CD45+% and CD3+% decreased, but CD4+ ICDB+ increased with statistical difference in high, middle, low dose WJR groups, and the Sis hen Pill control group (P <0. 05). Compared with the Sis hen Pill control group, there was statistical difference in all indices except CD45+ value in the low dose SWPSM group (P <0. 05). Compared with the low dose WJ R group, the expression of CD3+% decreased in high and middle dose WJR groups, and the Sis hen Pill control group; CD4+ /CD8+ increased in the Sishen Pill control group and the high dose SWPSM group (all P < 0. 05).</p><p><b>CONCLUSIONS</b>WJR showed better treatment effect. The mechanism of Shen warming Pi strengthening method might be achieved by regulating expressions of CD45+% and CD3+%, and CD4+ /CD8+ ratios.</p>

Significance of C-myc expression in T-lymphoblastic lymphoma/leukemia and its relation with prognosis / 中华病理学杂志

<p><b>OBJECTIVE</b>To study the C-myc gene and protein in T lymphoblastic lymphoma/leukemia (T-LBL/ALL) and its relationship to prognosis.</p><p><b>METHODS</b>60 cases of T-LBL/ALL with follow-up data were studied by using immunohistochemical EnVision method for CD1a, CD3, εCD3, CD7, CD10, CD34, CD43, CD45RO, CD99, TDT, CD20, CD23, MPO, Ki-67 and C-myc. 20 cases of reactive lymph nodes were selected as normal control group of C-myc gene and protein. Fluorescence in-situ hybridization (FISH) for C-myc gene (located on chromosome8q24) was performed to detect its breakage and gain.</p><p><b>RESULTS</b>Among the 60 cases of T-LBL/ALL, immunohistochemistry results showed:the percentages of tumor cells expression of CD1a, CD3, εCD3, CD7, CD10, CD34, CD43, CD45RO, CD99 and TDT were 38.3% (23/60), 75.0% (45/60), 45.0% (27/60), 95.0% (57/60), 36.7% (22/60), 23.3% (14/60), 60.0% (36/60), 41.7% (25/60), 96.7% (58/60) and 93.3% (56/60). Separately, while CD20, CD23 and MPO were all negative. A figure of Ki-67 expression ≤ 80% was found in 36 cases and > 80% was found in 24 cases. The positive rate of C-myc protein was 66.7% (40/60) in 60 cases of T-LBL/ALL, was 0% (0/20) in 20 cases of reactivated lymphoid tissue (χ² = 26.67, P < 0.05). C-myc protein expression was positively correlated with the mediatinal width and Ki-67 index (P < 0.05). Fluorescence in-situ hybridization results showed that among the 60 cases of T-LBL/ALL, C-myc gene with breakage of 8q24 was detected in 6 cases (10.0%), and gains in 11 cases (18.3%). 20 cases of reactive lymph nodes were not occurred breakage and gains of C-myc gene. It is not significant between C-myc gene and protein expression (P > 0.05). In addition, in 60 cases of T-LBL/ALL, 12(20.0%) cases of C-myc protein and genetic abnormalities coexist. Log-rank analysis results: The prognosis of C-myc protein positive group was worse than negative group (P < 0.05). The relationship of C-myc gene and prognosis was not significant (P > 0.05). C-myc protein and genetic abnormality coexist is related with worse prognosis (P < 0.05). COX analysis results show that the C-myc protein positive group may be a independent poor prognosis factors (P < 0.05).</p><p><b>CONCLUSIONS</b>C-myc may play an important role on the development of T-LBL/ALL. It may be a independent prognosis factors.</p>

Distribution of CD45RO⁺ T and CD45RA⁺ T Cells in Peripheral Blood of Patients with Peripheral T Cell Lymphoma / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the clinical significance of memory T cells (CD45RO⁺ T) and the initial T cells (CD45RA⁺ T) distribution in peripheral blood of patients with peripheral T-cell lymphoma (PTCL).</p><p><b>METHODS</b>A total of 27 patients diagnosed as PTCL in our hospital from February 2010 to February 2014 were collected in this study; whereas 30 healthy people were enrolled as control. The distribution of CD45RO⁺ T and CD45RA⁺ T cells were detected seperately in each group, and the results were analysed further.</p><p><b>RESULTS</b>The expression of T cell antigens in lymphnodes of PTCL patients were diverse: the CD4⁺ T cells were the main immune phenotype, while the B cell-related antigen was not expressed. The CD4⁺/CD8⁺, CD4⁺ CD45RO⁺ T in the peripheral blood of PTCL patients were significantly lower than that in normal group (P < 0.05); while the CD4, CD45RA⁺ T, CD8⁺ CD45RA⁺ T and CD8⁺ CD45RO⁺ T were significantly higher than that in normal group (P < 0.05). The patients in stage I/II had higher CD4⁺/CD8⁺, CD4⁺ CD45RO⁺ T than those in stage III/IV (P < 0.05), whereas the CD4⁺ CD45RA⁺ T, CD8⁺ CD45RA⁺ T and CD8⁺ CD45RO⁺ T were significantly lower than those in the stage III/IV patients (P < 0.05).</p><p><b>CONCLUSION</b>The distributions of CD45RA⁺ T and CD45RO⁺ T in PTCL patients are quite different, and the corresponding treatment might be adopted according to the different distribution of these cells, so as to improve the diagnosis and prognosis of PTCL.</p>

The characteristics of 62 cases of CD20-positive multiple myeloma / 中华血液学杂志

<p><b>OBJECTIVE</b>To explore the clinical and laboratory characteristics of mutiple myeloma patients with CD20 expression.</p><p><b>METHODS</b>Review the data of mutiple myeloma patients and analyze the clinical and laboratory characteristics of CD20 positive patients, compared with CD20 negative patients.</p><p><b>RESULTS</b>(1)Totally 465 cases of newly-diagnosed MM were collected with CD20 expression status detected by multi-color flow cytometry. Sixty two patients (13.3%) were CD20 positive and the others were negative. (2)No statistical differences were found between CD20 positive and negative groups about the sex ratio, age predominance, D-S staging, ISS staging, renal insufficiency rate, platelet count, LDH level and classifications by paraprotein(all P value>0.05). (3)Compared with those of CD20 negative patients, the hemoglobulin value(74.5 g/L vs 83.5 g/L, P=0.021), extramedullary involvement rate (3.5% vs 13.7%, P=0.029), CD56-positive rate(36.7% vs 68.8%,P=0.000), t(4;14)translocation rate(2.4% vs 24.0%, P=0.001) in CD20 positive patients were lower statistically. (4)Compared with those of CD20 negative patients, the percentage of plasma cells (0.400 vs 0.295, P=0.045) by marrow smear differential counting, the percentage of myeloma cells(20.0% vs 6.8%, P=0.000) by multi-color flow cytometry analysis, CD45-positive rate(12.1% vs 4.5%, P=0.018), CD79a-positive rate(9.8% vs 1.5%, P=0.013) and t(11;14) translocation rate(60.5% vs 14.4%, P=0.000)in CD20 positive patients were higher statistically. (5)There was no statistical differences about the overall response rate (ORR), complete response rate (CRR), TTP(time to progression), PFS(progression free survival) and overall survival (OS) between CD20 positive and negative groups.</p><p><b>CONCLUSION</b>CD20 positive rate is 13.3% in multiple myeloma pateints according to our data. CD20 poaitive myeloma were prone to residing in bone marrow and affecting erythropoiesis. Atypical immunophenotypes were more common, and the incidence of t(11;14) were increased markedly while that of t(4;14)were rare for CD20 positive multiple myeloma.</p>

Expression and clinical significance of CD45RO in laryngeal carcinoma tissue / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To investigate the role and significance of CD45RO in occurance and development in laryngeal squamous carcinoma, and to provide some valuable clues for searching new approaches to assess prognosis and theoretical basis for tumor biotherapy.@*METHOD@#The expression of CD45RO protein in 50 cases of laryngeal squamous carcinoma and 10 cases normal mucos was detected by immunohistochemical S-P method.@*RESULT@#The positive rate of CD45RO was 30% and 86% respectively in normal tissue and laryngeal squamous cell carcinoma tissue. The expresion of CD45RO was significantly and negatively associated with local metastatic of lymph nodes 0.713, P < 0.05) and tumor sites (r = -0.750, P < 0.05), but it have no notable difference with pathology differentiation, age, infiltrating depth and clinical stages in 50 cases of laryngeal squamous cell cancer.@*CONCLUSION@#(1) The expresion of CD45RO in laryngeal squamous cell cancer is more than that in normal tissue. (2) It is possible that overexpresion of CD45RO in laryngeal squamous cell carcinoma cut local metastatic lymph nodes. (3) It is probable that overexpresion of CD45RO in laryngeal squamous cell cancer made for prognosis of patients. (4) Other than UICC-TNM stage, pathology differentiation, it provide valuable clues for searching new approaches to assess prognosis of laryngeal squamous cell carcinoma.

Nodular histiocytic/mesothelial hyperplasia: a clinicopathologic analysis of 7 cases / 中华病理学杂志

<p><b>OBJECTIVE</b>To analyze the clinicopathologic and immunohistochemical features of nodular histiocytic/mesothelial hyperplasia (NHMH) and to improve the knowledge of this disease.</p><p><b>METHODS</b>Seven cases of NHMH were collected and the clinicopathologic and immunohistochemical data were analyzed with review of the literature.</p><p><b>RESULTS</b>Seven male patients aged from 1.5 to 5.0 years (mean 2.8). The main clinical symptom was an inguinal mass.Grossly, main pathological changes were the mural nodule or free nodule in lumen, with diameter of 0.1-0.5 cm.Histologically, the tumor cell morphology was relatively single, cohesive polygonal or oval cells which were arranged in solid sheets or nests, usually with ovoid or deeply grooved nuclei and a moderate amount of pale pink cytoplasm in the nodular collection area. The nuclei had delicate chromatin and no obvious atypia, and mitosis was incidentally found. A few scattered lymphocytes were found in the stroma. The cyst wall was lined by a single layer of mesothelial cells.Immunohistochemically, the most cells in nodular lesion were strongly positive for the histiocytic marker CD68, vimentin and α1-antichymotrypsin, while lining mesothelial cells on the wall were positive for calretinin, MC, WT1, CK5/6, CKpan and EMA.</p><p><b>CONCLUSIONS</b>NHMH is a rare and benign tumor-like lesion, and easy to be misdiagnozed, which should be distinguished from neuroendocrine tumors, Langerhans cell histiocytosis, seminoma, mesothelioma and so on. The correct diagnosis of this lesion depends on the clinical characteristics, morphology and immunohistochemistry.</p>

Immunophenotypic analysis of abnormal plasma cell clones in bone marrow of primary systemic light chain amyloidosis patients / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Primary systemic light chain amyloidosis (AL) is a rare plasma cell disease, our purpose was to analyze the immunophenotypic characteristics of the plasma cells in bone marrow in AL patients, and explore whether the detection of abnormal plasma cell clones in bone marrow by flow cytometry (FCM) could be used as an important indicator of AL diagnosis.</p><p><b>METHODS</b>Fresh bone marrow samples were collected from 51 AL, 21 multiple myeloma (MM), and 5 Waldenström's macroglobulinemia (WM) patients. The immunophenotype of bone marrow cells were analyzed and compared by FCM using a panel of antibodies including CD45, CD38, CD138, CD117, CD56, and CD19.</p><p><b>RESULTS</b>In AL, light chain restriction could be identified in 31 cases (60.9%), in which the λ light chain restriction was found in 24 cases (77.4%). In MM, κ light chain restriction was found in 13 cases (61.9%), and λ light chain restriction in eight cases. CD45 on abnormal plasma cells was negative to weakly positive in both AL and MM, but was positive to strongly positive in WM. In the bone marrow plasma cells of the 51 AL, 78.4% were CD56+, 68.6% were CD117+, and 88.2% were CD19-. While in the 21 MM cases, 66.7% were CD56+, 38.1% were CD117+, and 90.4% were CD19-. The plasmacytoid lymphocytes in the five WM patients were CD19+ and CD56-, CD117-.</p><p><b>CONCLUSION</b>Detection of abnormal plasma cell clones in bone marrow by FCM is valuable for the diagnosis of AL.</p>