OJECTIVE: To explore the changes of serum IgE and tryptase caused by anaphylactic shock rats and discuss the relation to PMI and preservative environment of corpse and specimen. METHODS: Rats were used for establishing anaphylactic shock models and randomly divided into room temperature group, refrigeration group, frozen group, manual hemolysis group, specimen preservation group. And the control group was also established. The blood samples were collected after rats were sacrificed. The degree of hemolysis was graded according to the color of the upper layer of the serum. The mass concentration of IgE and tryptase in each group was detected by ELISA. RESULTS: The levels of serum IgE and tryptase in anaphylactic shock dead rats were higher than that of the control group. Room temperature and frozen made obviously differences on the levels of serum IgE and tryptase with various PMI. The levels of serum IgE and tryptase in refrigeration group showed relatively stable. The levels of serum tryptase and IgE were elevated with differently increasing hemolysis. The levels of serum IgE and tryptase showed no obvious changes during the specimen kept under different temperature conditions for 25 days. CONCLUSION Serum IgE and tryptase obviously increased in anaphylactic shock rats. However, the levels were influenced by PMI and environmental temperature, especially under the conditions of room temperature and frozen.
Anaphylaxis/blood* ; Animals ; Disease Models, Animal ; Enzyme-Linked Immunosorbent Assay ; Immunoglobulin E/blood* ; Rats ; Temperature ; Tryptases/blood*

PURPOSE: The purpose of this study is to induce the differentiation of mast cells from human umbilical cord blood. METHODS: Mononuclear cells and CD34+ cells were obtained from cord blood and were cultured in the presence of stem cell factor, IL-3 and IL-6 in liquid suspension culture for 8 weeks. Mast cell was confirmed by Wright-Giemsa staining, immuno-histochemistry for tryptase and flowcytometry. RESULTS: When mononuclear cells were cultured for 4 weeks, the percentage of CD34-, CD117+ cells increased up to 8% in the presence of SCF only and 6.6% in the presence of SCF and IL-6. After 8 weeks of culture of CD34+ cells, the percentage of CD34-, CD117+ cells was highest at an average of 14.8% when cultured with SCF only, although absolute number of CD34-, CD117+ cells was higher when cultured in the presence of SCF, IL-3 and IL-6. CONCLUSION: We developed human mast cells from umbilical cord blood. However, some other factors such as combination or concentration of cytokines should be considered to enhance the efficiency of mast cell culture. In addition, mature cultured mast cells should be evaluated by flowcytometry as well as a special staining including immunohistochemistry.
Cytokines ; Fetal Blood* ; Humans* ; Immunohistochemistry ; Interleukin-3 ; Interleukin-6 ; Mast Cells* ; Stem Cell Factor ; Tryptases ; Umbilical Cord*

The detection of drug-induced anaphylactoid reactions remains a global challenge,still lacking mature and reliable animal models or test methods. Therefore,the purpose of this paper is to explore and establish the test methods and evaluation standards for anaphylactoid reactions that apply to injection drugs. Based on the anaphylactoid reaction symptoms of mice induced by intravenous injection drugs C48/40 and Tween 80,a list of systemic anaphylactoid reaction symptoms in mice was sorted out and an evaluation standard of anaphylactoid reactions symptoms was established by applying symptom intensity coefficient K( that can represent these verity of anaphylactoid reaction symptoms) and its calculation formula Accordingly,histamine,tryptase,and Ig E were selected as blood indicators of anaphylactoid reactions,so that a test method combining symptoms evaluation and blood makers detection was established.This test method could be used to evaluate the characteristics of anaphylactoid reactions: coefficient K,blood histamine levels were highly and positively correlated with C48/80 and Tween 80 dose; The log value of histamine was highly and positively correlated with K; tryptase level may rise,or remain steady,or drop,possibly associated with the characteristics of the tested object and time for blood taking; and Ig E level would drop or remain steady,but it would not rise,which can be clearly distinguished from type I allergic reactions. On this basis,tiohexol,iopromide,paclitaxel,Xuesaitong Injection,Shuanghuanglian Injection and Shengmai Injection were used to investigate the applicability. The testing results showed a high degree of consistency with the actual clinical situation. The results suggest that the method of systemic anaphylaxis test in mice has high sensitivity,specificity and good consistency with clinical practice.It is suggested to be further validated and popularized.
Anaphylaxis ; chemically induced ; diagnosis ; Animals ; Disease Models, Animal ; Drugs, Chinese Herbal ; toxicity ; Histamine ; blood ; Immunoglobulin E ; blood ; Injections, Intravenous ; Mice ; Shock ; chemically induced ; diagnosis ; Toxicity Tests ; Tryptases ; blood

OBJECTIVE: To investigate the relationship between tryptase in serum and anaphylaxis. METHODS: The concentrations of tryptase in the sera of heart blood in three persons died from anaphylaxis shock were detected by ELISA. The first sample was obtained from a man, aged 38, died of injecting Amikacin. The second sample was obtained from a man, aged 42, died of injecting Cephradine. The third sample was from a woman, aged 39, died of injecting Lincomycin. All samples were stored in -20 degrees C. RESULTS: The concentrations of tryptase in sera were 52 ng/ml, 121 ng/ml and 0.73 ng/ml. It was unknown why the concentration of tryptase in the third sample was normal. CONCLUSION In fetal anaphylaxia reaction tryptase measurement is a useful indicator, but the diagnosis is not to be based on the test alone.
Adult ; Anaphylaxis/enzymology* ; Biomarkers/blood* ; Cause of Death ; Enzyme-Linked Immunosorbent Assay ; Female ; Forensic Medicine ; Humans ; Male ; Mast Cells/enzymology* ; Postmortem Changes ; Retrospective Studies ; Serine Endopeptidases/blood* ; Tryptases

PURPOSE: Chronic rejection is the enemy in the battle for long term survival after renal allografts. Interstitial fibrosis is known to be the important finding in renal allografts with chronic rejection. Mast cells secrete a large number of fibrogenic factors and have been involved in chronic inflammation and tissue fibrosis. In this study the authors evaluated the relationship between mast cells and fibrosis in renal allografts with chronic rejection. METHODS: The authors evaluated 42 biopsied specimens of renal allografts. Immunohistochemistry using anti-mast cell tryptase (Dako, 1 : 200) and an LSAB kit (Dako) was applied to detect mast cells. The mean number of mast cells (MNM) per 10 high power fields was counted. RESULTS: MNM of implantation biopsies was 0.640+/-0.537, of acute rejection -1.969+/-1.216, of chronic rejection -6.0+/-3.133 (P<0.01), of acute tubular necrosis -1.360+/-0.899, and of acute cyclosporine nephrotoxicity -1.000+/-0.600. MNM according to donor source was 3.267+/-3.479 vs. 2.376+/-1.900 (living donors vs. cadaveric donors). MNM was significantly correlated with donor sex (male : female ratio of = 2.319+/-1.739 : 4.014+/-4.286, P<0.01), and cholesterol (hypercholesterolemia vs. non-hypercholesterolemia, 4.125+/-5.497 vs. 2.60+/-1.916, P<0.01). However, MNM according to blood pressure was not statistically significant (hypertension : non-hypertension ratio of=3.189+/-3.05 : 1.200+/-1.226, P>0.05). CONCLUSION: Our data show that the number of mast cells in renal allograft was significantly associated with chronic rejection, donor sex and hypercholesterolemia.
Allografts* ; Biopsy ; Blood Pressure ; Cadaver ; Cholesterol ; Cyclosporine ; Female ; Fibrosis ; Humans ; Hypercholesterolemia ; Immunohistochemistry ; Inflammation ; Kidney Transplantation ; Mast Cells* ; Necrosis ; Tissue Donors ; Tryptases

OBJECTIVEMast cells and eosinophil have been found to play important roles not only in the development of anaphylactic inflammation but also in the chronic progression of organ reconstruction. But their role in the pathogenesis of Henoch-Schonlein purpura nephritis (HSPN) has not been fully understood. The present study was conducted to observe the serum levels of eosinophil cationic protein (ECP) and renal infiltration of mast cells in HSPN in order to elucidate their role in the development and progression of HSPN in children.

METHODSThe serum ECP levels were determined in 46 children with HSPN by fluoro-enzyme immunoassay (FEIA) using the Pharmacia CAP System. The distribution of mast cells infiltration was detected by immuno-enzyme-histological staining of tryptase (a marker for mast cell activation) and their relation with pathological changes was analyzed in 32 children with HSPN.

RESULTSThe serum ECP levels were 16.3 +/- 6.5 microg/L in the active stage of HSPN, significantly higher than that in remission stage (3.9 +/- 1.4 microg/L, P < 0.01) and that in control group (3.1 +/- 1.7 microg/L, P < 0.01). The number of mast cells in renal interstitium was 4.4 +/- 2.4 cells/mm2 in normal kidney, and significantly increased to 27.2 +/- 19.2 cells/mm2 in children with HSPN ISKDC grade II (P < 0.01) and 42.1 +/- 16.4 cells/mm2 in grade III (P < 0.05 when compared with grade II), 77.9 +/- 15.0 cells/mm2 in grade IV (P < 0.05 when compared with grade III).

CONCLUSIONThe serum ECP level could reflect disease activity of HSPN, and mast cell infiltration in kidney correlated significantly with renal histological severity in HSPN. Mast cells and eosinophil may play important roles in the development and progression of HSPN.

Biomarkers ; blood ; Child ; Disease Progression ; Eosinophil Cationic Protein ; blood ; Female ; Fluoroimmunoassay ; Humans ; Immunohistochemistry ; Kidney ; metabolism ; pathology ; Male ; Mast Cells ; metabolism ; pathology ; Nephritis ; blood ; metabolism ; pathology ; Purpura, Schoenlein-Henoch ; blood ; metabolism ; pathology ; Severity of Illness Index ; Tryptases ; metabolism

OBJECTIVE: To increase the death rate of fatal anaphylaxis in guinea pigs and the detectahie level of the tryptase of mast cell in hlood serum. METHODS: Seventy-four guinea pigs were randomly divided into five groups: original model group, original model control group, improved model group, improved model control group, improved model with non-anaphylaxis group. Using mixed human serum as the allergen, the way of injection, sensitization and induction were improved. ELISA was used to detect the serum mast cell tryptase and total IgE in guinea pigs of each group. RESULTS: The death rate of fatal anaphylaxis in original model group was 54.2% with the different degree of hemopericardium. The severe pericardial tamponade appeared in 9 guinea pigs in original model group and original model control group. The death rate of fatal anaphylaxis in improved model group was 75% without pericardial tamponade. The concentration of the serum total IgE showed no statistically difference hetween original model group and original model control group (P > 0.05), hut the serum mast cell tryptase level was higher in the original model group than that in the original model control group (P > 0.05). The concentration of the serum total IgE and the serum mast cell tryptase level were significantly higher in improved model group than that in the improved model control group (P < 0.05). CONCLUSION The death rate of the improved model significantly increases, which can provide effective animal model for the study of serum total IgE and mast cell tryptase.
Allergens/immunology* ; Anaphylaxis/pathology* ; Animals ; Cause of Death ; Disease Models, Animal ; Enzyme-Linked Immunosorbent Assay ; Forensic Pathology ; Guinea Pigs ; Humans ; Immunoglobulin E/blood* ; Larynx/pathology* ; Lung/pathology* ; Male ; Mast Cells/immunology* ; Serum/immunology* ; Tryptases/blood*

OBJECTIVETo investigate the effect of sodium cromoglycate on experimental endometriosis in rats.

METHODSEndometriosis model was established in 36 unpregnant female SD rats by transplanting autologous fragments of endometrium to the inner surface of the abdominal wall. The endometriotic lesions were measured by a second laparotomy 2 weeks after surgery. Then the rats were randomly divided into four groups (n=8 in each group) to receive intraperitoneal injection of different doses of sodium cromoglycate for 2 weeks: high-dose group (20 mg·kg⁻¹·d⁻¹); low-dose group (10 mg·kg⁻¹·d⁻¹); the negative control group and the blank control group. The animals were sacrificed and the size of the lesions were measured. The endometriosis model of SD rats was identified by HE staining and immunohistochemical staining of keratin and vimentin. The total number of mast cells and their degranulation were measured by Toluidine blue staining; the concentrations of TNF-α in serum were measured by enzyme linked immunosorbent assay; the concentrations of estradiol in serum were measured by enzyme immunoassay; the expression of tryptase and nerve growth factor (NGF) were measured by immunohistochemical staining.

RESULTSThe number of activated mast cells (MC) by Toluidine blue staining in high-dose group was significantly lower than that in negative control group (P<0.05), and its ratio of degranulation/total number of MC was significantly lower than that in negative control group or blank control group (P<0.05). The serum TNF-α levels and tryptase expression in tissues in high-dose group were significantly lower than those in negative control group or blank control group (P<0.05). However, no significant difference in the size of endometriotic lesions and expression of NGF was found among groups (P>0.05).

CONCLUSIONSodium cromoglycate can stabilize mast cells from degranulation, which may relieve the clinical symptoms of endometriosis by reducing TNF-α and tryptase levels.

Animals ; Cromolyn Sodium ; pharmacology ; Disease Models, Animal ; Endometriosis ; drug therapy ; Endometrium ; pathology ; Female ; Mast Cells ; drug effects ; Nerve Growth Factor ; metabolism ; Rats ; Rats, Sprague-Dawley ; Tryptases ; metabolism ; Tumor Necrosis Factor-alpha ; blood

BACKGROUNDAtherosclerosis (AS) is an inflammatory disease. Inflammation was considered to play a role in the whole process of AS. This study aimed to analyze the relationships of inflammatory factors and risk factors with different target organ damages (TOD) in essential hypertension (EH) patients and to explore its clinical significance.

METHODSA total of 294 EH patients were selected and divided into four groups according to their conditions of TOD. Forty-eight healthy subjects were selected as control. The clinical biochemical parameters, serum amyloid A, serum tryptase, and lipoprotein-associated phospholipase A2 (Lp-PLA2) in each group were detected, and the related risk factors were also statistically analyzed.

RESULTSFibrinogen (Fbg) was the most significant independent risk factor in acute coronary syndrome (ACS) group (odds ratio [OR]: 22.242, 95% confidence interval [CI]: 6.458-76.609, P< 0.001) with the largest absolute value of the standardized partial regression coefficient B' (b': 1.079). Lp-PLA2 was the most significant independent risk factor in stroke group (OR: 13.699, 95% CI: 5.236-35.837, P< 0.001) with b' = 0.708. Uric acid (UA) was the most significant independent risk factor in renal damage group (OR: 15.307, 95% CI: 4.022-58.250, P< 0.001) with b' = 1.026.

CONCLUSIONSFbg, Lp-PLA2, and UA are the strongest independent risk factors toward the occurrence of ACS, ischemic stroke, and renal damage in EH patients, thus exhibiting the greatest impacts on the occurrence of ACS, ischemic stroke, and renal damage in EH patients, respectively.

1-Alkyl-2-acetylglycerophosphocholine Esterase ; Aged ; Antihypertensive Agents ; therapeutic use ; Enzyme-Linked Immunosorbent Assay ; Essential Hypertension ; blood ; complications ; drug therapy ; physiopathology ; Female ; Humans ; Kidney Diseases ; blood ; etiology ; physiopathology ; Logistic Models ; Male ; Middle Aged ; Renal Insufficiency, Chronic ; blood ; etiology ; physiopathology ; Risk Factors ; Serum Amyloid A Protein ; metabolism ; Stroke ; blood ; etiology ; physiopathology ; Tryptases ; blood

AIMTo investigate the role of fexofenadine, a mast cell blocker, on semen quality in the treatment of infertile men.

METHODSThe study included 16 Turkish idiopathic infertile men with azoospermia or oligozoospermia who underwent testicular biopsy to examine mast cells containing tryptase. In all patients, a complete medical history, clinical examination, semen analysis and serum hormone assay were carried out. The biopsy specimens were immunohistochemically stained with antihuman tryptase for mast cells. The number of total mast cells per seminiferous tubule was calculated and recorded as mast cell index. The patients were divided into two groups according to their mast cell index: the higher (> or =1, n=9) and the lower (<1, n=7) index groups. Fexofenadine was administered orally at a dose of 180 mg/day for 4 to 9 months. Pre- and post-treatment semen parameters, including total motile sperm counts (TMC) were recorded and compared. Spontaneous pregnancies after the treatment were registered.

RESULTSThere was no statistically significant difference in TMC between the pre-treatment and post-treatment values in patients with higher and lower mast cell index (P> or =0.05). In both groups, nobody had a significant response to the treatment and there was no spontaneous pregnancy after the treatment.

CONCLUSIONAlthough testicular dysfunction is closely associated with increased number of testicular mast cells, fexofenadine, a mast cell blocker, appears not having any benefit in the treatment of Turkish infertile men with a significant increase in testicular mast cells.

Adult ; Biopsy, Needle ; Follicle Stimulating Hormone ; blood ; Histamine H1 Antagonists ; therapeutic use ; Humans ; Infertility, Male ; drug therapy ; pathology ; Male ; Mast Cells ; drug effects ; enzymology ; pathology ; Middle Aged ; Semen ; drug effects ; physiology ; Serine Endopeptidases ; metabolism ; Sperm Count ; Sperm Motility ; drug effects ; Terfenadine ; analogs & derivatives ; therapeutic use ; Testis ; anatomy & histology ; pathology ; Tryptases ; Turkey