Expression of CD63 in Lung Tissue of Guinea Pigs Dying of Anaphylactic Shock / 法医学杂志

Objective To study the protein expression of cluster of differentiation 63 （CD63） in lung tissues of guinea pigs that died of anaphylactic shock and discuss the diagnostic value of CD63 for death from anaphylactic shock. Methods Twenty guinea pigs were randomly divided into control group, anaphylactic shock immediate death group, cold storage group （4 ℃ for 48 h） and frozen group （-20 ℃ for 7 d）. The animal model of guinea pigs that died of anaphylactic shock was established with human mixed serum injection. The expression changes of CD63 protein and CD63 mRNA in lung tissues were detected by hematoxylin-eosin （HE） staining, immunohistochemical staining, Western blotting, enzyme-linked immunosorbent assay （ELISA） and real-time RT-PCR. Results HE staining results showed congestion, and edema of lung tissues, and eosinophil infiltration in the anaphylactic shock groups. Western blotting analysis results showed that the expression of CD63 protein in the lung tissues of guinea pigs that died of anaphylactic shock was significantly higher than that in the control group （P<0.05）. Comparison between the anaphylactic shock groups was made, and the differences had no statistical significance. The results of immunohistochemical staining and real-time RT-PCR were consistent with that of Western blotting. ELISA results showed that CD63 protein expression in the immediate death group was higher than that in the control group （P<0.05）. Conclusion The expression of CD63 protein and CD63 mRNA in the lung tissues of guinea pigs that died of anaphylactic shock is significantly enhanced. Animal carcasses which were put in cold storage for 48 h and frozen for 7 d do not affect the examination of the above indicators. CD63 protein is expected to become an auxiliary diagnostic indicator of death from anaphylactic shock.

The apoptotic, angiogenic and cell proliferation genes CD63, S100A6 e GNB2L1 are altered in patients with endometriosis / Os genes apoptóticos, angiogênicos e de proliferação celular CD63, S100A6 e GNB2L1 estão alterados em pacientes com endometriose

Abstract Objective The aim of the present study was to analyze the expression of the CD63, S100A6, and GNB2L1genes, which participate in mechanisms related to the complex pathophysiology of endometriosis. Methods A case-control study was conducted with 40 women who were diagnosed with endometriosis, and 15 fertile and healthy women. Paired samples of eutopic endometrium and endometriotic lesions (peritoneal and ovarian endometriotic implants) were obtained from the women with endometriosis in the proliferative (n = 20) or secretory phases (n = 20) of the menstrual cycle. As controls, paired endometrial biopsy samples were collected from the healthy women in the proliferative (n = 15) and secretory (n = 15) phases of the samemenstrual cycle.We analyzed the expression levels of the CD63, S100A6, and GNB2L1 genes by real-time polymerase chain reaction. Results An increase in CD63, S100A6, and GNB2L1 gene transcript levels was observed in the ectopic implants compared with the eutopic endometrium of the women with and without endometriosis, regardless of the phase of the menstrual cycle. Conclusion These findings suggest that the CD63, S100A6, and GNB2L1 genesmay be involved in the pathogenesis of endometriosis, since they participate in mechanisms such as inhibition of apoptosis, angiogenesis and cell proliferation, which lead to the loss of cell homeostasis in the ectopic endometrium, thus contributing to the implantation and survival of the tissue in the extrauterine environment.

Resumo Objetivo O objetivo do presente estudo foi analisar a expressão dos genes CD63, S100A6 e GNB2L1, que participam em mecanismos relacionados à complexa fisiopatologia da endometriose. Métodos Um estudo caso-controle foi realizado com 40 mulheres diagnosticadas com endometriose e 15 mulheres férteis e saudáveis. Amostras pareadas de endométrio eutópico e de lesões endometrióticas (implantes endometrióticos peritoneais e ovarianos) foram obtidas de mulheres com endometriose nas fases proliferativa (n = 20) ou secretora (n = 20) do ciclo menstrual. Como controle, amostras pareadas de biópsia endometrial foram coletadas de mulheres saudáveis nas fases proliferativa (n = 15) e secretora (n = 15) nomesmo ciclomenstrual. Foram analisados os níveis de expressão dos genes CD63, S100A6 e GNB2L1 por reação em cadeia da polimerase em tempo real. Resultados Foi observado um aumento nos níveis de transcritos dos genes CD63, S100A6 e GNB2L1 em implantes ectópicos quando comparado ao endométrio eutópico de mulheres com e sem endometriose, independente da fase do ciclo menstrual. Conclusão Estes achados sugerem que os genes CD63, S100A6 e GNB2L1 podem estar envolvidos na patogênese da endometriose, pois participam de mecanismos como inibição de apoptose, angiogênese e proliferação celular, os quais levam à perda da homeostase celular no endométrio ectópico e, portanto, contribuem para o implante e a sobrevivência do tecido no ambiente extrauterino.

Effect of clopidogrel on platelet CD markers in normal individuals and in patients with untreated type 2 diabetes mellitus

The objective of this study was to evaluate the effect of clopidogrel response in patients with untreated type 2 diabetes mellitus as compared with normal individuals. One hundred and seven subjects i.e. 32 normal and 75 patients with untreated type 2 diabetes mellitus were enrolled in this study. In the first step, normal subjects as well as diabetic patients were selected and tested for various laboratory parameters and platelets flow cytometry. In the second step, an antiplatelet drug [clopidogrel] was administered for 10 days to each individual enrolled in the study. After 10 days blood samples were collected for platelets flow cytometry. CD41 and CD61 did not show any change after the administration of clopidogrel in resting and activated platelets. CD63 and CD62p positivity was increased in normal and in diabetic patients' platelets after activation with ADPbefore clopidogrel. It was decreased in normal resting and ADPstimulated platelets after clopidogrel treatment. CD63 and CD62p positivity in resting and ADP stimulated patients' platelets was also decreased after clopidogrel treatment. The change was, however, not as marked as in normal subjects

Diagnostic Significance of BAT in Anaphylaxis to Non-ionic Contrast Media / 法医学杂志

OBJECTIVE@#To investigate the diagnostic significance of basophil activation test (BAT) in anaphylaxis to non-ionic contrast media through testing the content of CD63, mast cell-carboxypeptidase A3 (MC-CPA3), and terminal complement complex SC5b-9 of the individuals by testing their levels in the normal immune group and the anaphylaxis groups to β-lactam drugs and non -ionic contrast media.@*METHODS@#The CD63 expression of basophilic granulocyte in blood was detected by flow cytometry. The levels of MC-CPA3 in blood serum and SC5b-9 in blood plasma were detected by ELISA.@*RESULTS@#The CD63 expression of basophilic granulocyte in blood, the levels of MC-CPA3 and SC5b-9 of anaphylaxis to non-ionic contrast media and β-lactam drugs were significantly higher than that in normal immune group (P < 0.05).@*CONCLUSION@#There is activation of basophilic granulocytes, mast cells and complement system in anaphylaxis to non-ionic contrast media. BAT can be used to diagnose the anaphylaxis to non-ionic contrast media.

Immunologic Evaluation of Immediate Hypersensitivity to Cefaclor

PURPOSE: Cefaclor is widely prescribed for various infectious diseases. As its consumption increases, the number of hypersensitivity reactions to cefaclor has increased. This study aimed to evaluate the immunologic findings of immediate hypersensitivity to cefaclor. MATERIALS AND METHODS: We enrolled 47 patients with immediate hypersensitivity to cefaclor from Ajou University Hospital and Asan Medical Center. Serum specific IgE, IgG1, and IgG4 antibodies to cefaclor-human serum albumin (HSA) conjugate were measured by enzyme-linked immunosorbent assay (ELISA). RESULTS: The most common phenotype was anaphylaxis (Group I, 78.7%), followed by urticaria (Group II, 21.3%). The detection of specific IgE, IgG1, and IgG4 to cefaclor-HSA conjugate by ELISA tended to be higher in Group I (40.5%, 41.7%, 21.6%) than in Group II (20.0%, 20.0%, 0%) with no statistical significance. Significant associations were found between specific IgE and IgG1 or IgG4 (p<0.001, p=0.019). ELISA inhibition tests showed significant inhibitions by both free cefaclor and cefaclor-HSA conjugate. For basophil activation tests in patients having no specific IgE antibody, the CD63 expression level on basophils increased with incubations of free cefaclor. CONCLUSION: The most common manifestation of immediate hypersensitivity to cefaclor was anaphylaxis, most of which was mediated by IgE; however, a non-IgE mediated direct basophil activation mechanism was suggested in a subset of anaphylaxis patients.

Basophil CD63 expression in the blood of the anaphylactic shock rat / 法医学杂志

OBJECTIVE@#To explore the value of flow cytometry in anaphylactic shock diagnosis by CD63 expression being detected using flow cytometry to conform the activation of basophils.@*METHODS@#Sixteen rats were randomly divided into two groups: control group and anaphylactic shock group. The model of anaphylactic shock rat with ovalbumin injection was established. CD63, CD45 and CD203c antibody combination, flow cytometry was employed to detected blood basophil CD63 expression. Immunofluorescence method was employed to observe the CD63 immunofluorescence staining in the rat lung tissue.@*RESULTS@#(1) Pure basophils were obtained by CD45 and CD203c gating. (2) The percentages of basophils CD63 were (17.34 +/- 2.04)% and (1.52 +/- 0.35)% in the experimental and control group, respectively. The differences between two groups were statistically significant (P < 0.01). (3) Compared with the control group, the expression of CD63 in basophils increased in anaphylactic shock lung tissue.@*CONCLUSION@#The detection of CD63 by flow cytometry could be the supplement of vivo allergic reactions and have good clinical value.

Real-time detection of mast cell degranulation in anaphylactoid reaction / 中国中药杂志

<p><b>OBJECTIVE</b>To establish a new, real time, dynamic and direct optical detection method for mast cell degranulation caused by anaphylactoid reaction.</p><p><b>METHOD</b>A CD63-GFP plasmid was constructed and introduced steadily into rat basophilic leukemia (RBL-2H3) cells. The movements of CD63-GFP, which was located on both the granule membranes and the plasma membranes of RBL cells stimulated by Compound 48/80, were studied by confocal laser scanning microscope (CLSM) and total internal reflection fluorescence microscope (TIRFM) both inside and on the surface of living RBL-2H3 cells.</p><p><b>RESULT</b>Before antigen stimulation, most granules with CD63-GFP hardly moved in RBL cells. However, after antigen stimulation, the granules moved dramatically. They reached the plasma membranes in a few minutes and fused with them instantaneously. The velocity of the granule movement toward the plasma membranes on antigen stimulation was calculated to be 0.05 micron x s(-1).</p><p><b>CONCLUSION</b>Analysis of the movement of each granule provided a new insight into the elementary process of degranulation. The method is rapid, sensitive and reliable, which could be used as a new detection method for anaphylactoid reaction in vitro.</p>

Significance of serum CD62p and CD63 levels in patients with head injury / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To determine the serum levels of CD62p (alpha-granular membrane protein) and CD63 (lysosome intact membrane protein) in patients with head injury and to observe its relation to injury severity.</p><p><b>METHODS</b>Fifty-three patients with head injury were divided into 3 groups; Group A patients with mild head injury; Group B with moderate head injury; and Group C with severe head injury. The serum levels of CD62p, CD63 were measured on 12 h, d 1, 3, 5 and 7 after injury.</p><p><b>RESULT</b>The serum levels of CD62p and CD63 in Group B and Group C were higher than those in Group A and control (P<0.05). The serum level of CD62p in Group C was higher than that in Group B (P<0.05). The serum levels of CD62p in Group C on d 1, 3, 5 after injury were higher than those on 12 h (P<0.05). The serum level of CD63 in Group B on d 3 after injury were higher than that on 12 h (P<0.05). The serum levels of CD63 in Group C on d 1, 3, 5 after injury were higher than those on 12 h (P<0.05).</p><p><b>CONCLUSION</b>The serum levels of CD62p and CD63 in patients with head injury may be helpful for identifying the severity of injury, and CD62p seems to be more sensitive.</p>

The relationship between the peripheral blood of CD61, CD63, PAC-1 and the transplant kidney function / 中华外科杂志

<p><b>OBJECTIVES</b>To explore the relationships between the peripheral blood levels of CD61, CD63, PAC-1 and the incidence of acute rejection and tubular necrosis after renal transplantation, and recovery of the graft function.</p><p><b>METHODS</b>The peripheral blood levels of CD61, CD63, and PAC-1 of 86 patients with uremia in different stages before and after transplantations were analyzed by flow cytometry. The patients were divided into three groups: (1) twenty-nine patients with normal grafts function, (2) hirty with acute rejection and (3) twenty-seven with acute tubular necrosis. The patients with acute rejection were randomly divided into treatment group with anticoagulants and cntrol group.</p><p><b>RESULTS</b>The peripheral blood levels of CD61, CD63 and PAC-1 significantly increased (P < 0.05) in the patients with acute rejection, in comparison with those with normal grafts function and those with acute tubular necrosis. The peripheral blood levels of CD61, CD63 and PAC-1 in patients with acute rejection in anticoagulants therapy was lower, recovery time of the grafts function was shorter, one-year survival rates of patients and grafts were higher, as compared with those of controls.</p><p><b>CONCLUSIONS</b>The patients with acute rejection have significantly high peripheral blood levels of CD61, CD63 and PAC-1 before transplantation, however, these values in patients with acute tubular necrosis are not high, this suggesting that acute rejection might relate to platelet activation, while acute tubular necrosis might not relate to it. After anticoagulants therapy in patients with acute rejection, the grafts function might recover faster and their one-year survival rates and grafts might be higher in those with CD61, CD63 and PAC-1 decreasing remarkably.</p>

Detection of platelet Ca2+(i), CD62P, CD63 and plasma CD62P in cirrhosis patients / 中华肝脏病杂志

<p><b>OBJECTIVES</b>To study on the relationship between platelet Ca2+(i), CD62P, CD63, serum CD62P (SCD62P) and cirrhosis patients.</p><p><b>METHODS</b>Platelet CD62P, CD63 were determined with flow cytometry, SCD63P with ELISA, and Ca2+(i) in platelet was determined with fluorophotometry.</p><p><b>RESULTS</b>Platelet Ca2+(i), CD62P, CD63, and SCD62P levels in cirrhosis patients were (103.1+/-22.2)nmol/L, (47.6+/-20.0)%, (47.1+/-24.6)%, and (67.6+/-37.6)microg/L, and in controls were (57.6+/-13.1)nmol/L, (3.1+/-0.7)%, (2.5+/-0.7)%, and (24.0+/-6.5)microg/L, respectively. The levels in the former were higher than those in the latter (t > or = 6.148, P<0.05). The above levels in upper gastrointestinal haemorrhage group were much higher than those in the non-haemorrhage group (120.3nmol/L+/-18.8nmol/L vs 91.1nmol/L+/-14.3nmol/L, 64.9%+/-14.7% vs 34.6%+/- 11.9%, 70.9%+/-14.5% vs 30.2%+/-14.4%, and 103.6microg/L+/-14.9microg/L vs 40.8microg/L+/-24.0microg/L, respectively, t > or = 5.380, P<0.05). But the numbers of platelet between the two groups were no obvious difference.</p><p><b>CONCLUSIONS</b>Platelet in the cirrhosis patients is greatly active, and the detection of platelet CD62P, CD63, SCD62P has a certain value in judging the degree of cirrhosis.</p>