The differential roles of caspase family members in mediating PF4-induced breast cancer apoptosis

@#Introduction: This study focused on PF4 effects on caspase-3,-6, -7, -8 and -9 which regulate the apopotosis process in breast cancer. Materials and Methods: Breast tumours were induced in forty 21-day-old female Sprague Dawley rats (SDRs) using MNU until tumour size reached 14.5 mm (SD: 0.5 mm). The rats were then divided into two groups: Group 1 (control injected with 0.9% saline; n = 20), and Group 2 (platelet factor 4 (PF4); n = 20). PF4 was administered through focal intralesional injection at 20 µg/lesion dose. Following 5-day treatment, the SDRs were sacrificed. Subsequently, representative sections from the tumour were obtained for haematoxylin and eosin (H&E) staining. The expressions of caspase-3, -6, -7, -8 and -9 were evaluated using immunohistochemistry (IHC) staining. Results: The majority of breast tumour specimens were of aggressive types [ncontrol = 13 (65%); nPF4 = 12 (60%)]. Invasive ductal carcinoma not otherwise specified (IDC-NOS) was the most commonly observed breast tumour histology for control and PF4 groups (n = 8 (40%) in respective groups). PF4-treated group exhibited significant differences in the caspase-3, -6 and -8 expression levels compared to the control group (all p < 0.001). There were no significant differences in caspase-7 (p = 0.347) and caspase-9 (p = 0.373) expression levels between both groups. Conclusion: This study found that PF4 acts via the caspase-mediated extrinsic apoptosis pathway without the involvement of the intrinsic pathway.

Tratamiento exitoso con fondaparinux en trombocitopenia inducida por heparina y trombosis / Successful treatment with fondaparinux in heparin-induced thrombocytopenia and thrombosis

La trombocitopenia inducida por heparina (TIH) es una reacción adversa inmunológica caracterizada por trombocitopenia y la asociación paradojal de trombosis arterial o venosa. Es causada por la formación de anticuerpos IgG contra el complejo multimolecular de heparina-factor plaquetario 4 (FP4). Fondaparinux es un inhibidor selectivo del factor Xa que tiene escasa afinidad por el FP4 y posee un menor potencial para inducir una respuesta inmunológica, haciendo del mismo un agente potencialmente útil en el tratamiento de la TIH. Se presenta el caso de una mujer de 73 años con TIH asociada a fenómenos trombóticos arteriales y venosos, que recibió exitosamente fondaparinux, con normalización del recuento plaquetario y sin progresión trombótica.

Heparin induced thrombocytopenia (HIT) is an immune-mediated adverse reaction characterized by thrombocytopenia and paradoxical arterial or venous thrombosis, due to the formation IgG antibodies directed to a multimolecular complex of heparin-platelet factor 4 (PF4). Fondaparinux is a selective factor Xa inhibitor with little affinity for PF4 and thus less likely to induce an immune response, making fondaparinux a potentially useful drug for the treatment of HIT. Herein we report the case of a 73 years old woman with HIT associated with arterial and venous thrombosis that was successfully treated with fondaparinux, with normalization of the platelet countand without progression of thrombosis.

Effects of simvastatin on lung tissue angiogenesis and the gene expression of VEGF and PF4 of rats with pulmonary fibrosis / 实用医学杂志

Objective The study was designed to observe influence of simvastatin on lung tissue angiogenesis and the gene expression of vascular endothelial growth factor(VEGF) and platelet factor 4(PF4) of rats with bleomycin (BLM)-induced pulmonary fibrosis. Methods Ninety-six healthy male SD rats were divided into four groups by random number table, including normal control group (A), bleomycin group (B), prednisone acetate treatment group (C) and simvastatin treatment group (D). Lung tissue of rats in each group was detected as specimens. HYP was detected by digestion method. Angiogenesis, VEGF and PF4 protein expression were determined by immunohistochemical method (SP). Expression of VEGF and PF4 mRNA were respectively detected by RT-PCR assay. Results (1)HYP content of group C, D was lower than the group B, which was statistical significance (P <0.01). (2)MVD and the expression of VEGF in group B, C and D was higher than that in group A. PF4 expression of group B, C and D were lower than that of group A (P < 0.01). MVD and the expression of VEGF of group D were lower than those of group B, the expression of PF4 of group D was higher than that in group B (P < 0.05). Conclusion Mechanism of simvastatin on pulmonary fibrosis may be related to regulate the expression of VEGF and PF4 in lung tissue, inhibit pathological angiogenesis.

Application of ELISA in Detecting Heparin-Induced Thrombocytopenia Antibodies / 现代检验医学杂志

Objective To develop an ELISA method for determination of heparin-induced thrombocytopenia (HIT)antibody. Methods The compound formed between human platelet factor 4 (PF4)and heparin was used as the coating antigen,incu-bating the patients plasma with the coating antigen in the well,after washing,the second antibody labeled HRP was added in the well to incubate and washing again,the chromogenic substrates was added in the well to incubate,when the stop reaction was finished,the absorbance A450/A630 was detected,and the test results were judged according to standard,this method was compared with IBL method and was optimized and evaluated the performance.Results An indirect ELISA method was de-velop with the purified human PF4,the optimal dilution of sample and second antibody were 1∶100 and 1∶1 500 which de-tected by the orthogonal test,the intra-and inter-assay average coefficients of variation were 7.66% and 7.76%(<10%) respectively that detected by repeated measurement the three positive standard plasma.Through measureing the 100 healthy human plasm with no history of using heparin,the positive and negative predictive reference values were 0.304 and 0.456. IBL and this method detected 100 hemodialysis patients samples at the same time,and the result of statistical analysis was that,the sensitivity,speciality and accuracy of this method were 90%,97.78% and 90%,respectively.The negative and posi-tive predictive value were 81.8% and 98.88% respectively,and the difference was statistically significant [K=0.84(0.81~1)and Pexac=0.012<0.05].The difference was statistically significant,consistency was optimal,95% confidence interval was 92.59%~92.59%.Conclusion Comparing with the IBL,the method reported by this article had the similar perform-ance and good consistency,and it could satisfy the clinical detection and diagnosis of HIT patients.

Positive rate of PF4/H antibody, its risk factors and association with thrombosis in maintenance hemodialysis patients / 第二军医大学学报

Objective To study the positive rate of PF4/H antibodies in maintenance hemodialysis patients, and to analyze its risk factor and association with thrombosis. Methods A total of 157 maintenance hemodialysis patients were recruited in the present study. A crose-sectional and a longitude study were designed, and statistical analysis was used to analyze the risk factors of PF4/H antibody level and its association with thrombosis event. Results We found that 40.8% (64/ 157) of the patients were positive for PF4/H antibody. Past-thrombosis events, heparin types, duration and weekly dialysis hours were significantly different between antibody-positive group and antibody-negative group (P<0.05). Incidence of thrombosis event in antibody-positive group was significantly lower than that in the antibody-negative group (P<0.05). The risk ratio of antibody positivity for thrombosis event was 2.349. The incidence of thrombotic event in antibody-positive patients who took anti-platelet agents was lower than that in those who did not take. Conclusion The positive rate of PF4/H antibody has been found to be 40.8% in the present group. Thrombosis events, heparin types, duration and weekly dialysis hours are the risk factors of PF4/H antibody level. PF4/H antibody can serve as a marker for thrombotic events, and anti-platelet agents are effective for preventing from thrombotic events in PF4/H antibody positive patients.

Positive rate of PF4/H antibody, its risk factors and association with thrombosis in maintenance hemodialysis patients / 第二军医大学学报

Objective To study the positive rate of PF4/H antibodies in maintenance hemodialysis patients, and to analyze its risk factorand association with thrombosis. Methods A total of 157 maintenance hemodialysis patients were recruited in the present study. A cross-sectional and a longitude study were designed, and statistical analysis was used to analyze the risk factors of PF4/H antibody level and its association with thrombosis event. Results We found that 40. 8% (64/ 157) of the patients were positive for PF4/H antibody. Past-thrombosis events, heparin types, duration and weekly dialysis hourswere significantly different between antibody-positive group and antibody-negative group (P<0. 05). Incidence of thrombosis event in antibody-positive group was significantly lower than that in the antibody-negative group(P<0. 05). The risk ratio of antibody positivity for thrombosis event was 2. 349. The incidence of thrombotic event in antibody-positive patients who took anti-platelet agentswas lower than that in those who did not take. Conclusion The positive rate of PF4/H antibody has been found to be 40. 8% in the present group. Thrombosis events, heparin types, duration and weekly dialysis hours are the risk factors of PF4/H antibody level. PF4/H antibody can serve as a marker for thrombotic events, and anti-platelet agents are effective for preventing from thrombotic events in PF4/H antibody positive patients.

The Effect of Saffron in Patients with Autoimmune Diseases / 日本東洋医学雑誌

Saffron (<i>Crocus sativus</i> L.) is classified as a beneficial herb in the treatment of “Oketsu,” eliminating blood stagnation. The clinical symptoms of “Oketsu” include feeling cold and arthralgia. The pathological condition of “Oketsu” is considered to be increased blood viscosity and microvascular disorders.However, no useful biomarker has been reported for evaluation of the degree of “Oketsu.” Here, we investigated the clinical effect of saffron on the symptoms of “Oketsu” in patients with autoimmune diseases. At the same time, we measured the plasma levels of platelet factor 4 (PF-4) and beta-thromboglobulin (β-TG) as platelet activation markers. Seventy-one patients (66 women and 5 men, mean age 52.3 ± 16.1) were studied. They were administered saffron (300mg∼900mg) with traditional Kampo medicine. The clinical symptoms of “Oketsu” improved (80.9%, n=38/47) and we measured PF-4 and β-TG in pre-and post-saffron treatment periods. The plasma levels of PF-4 and β-TG significantly decreased after saffron therapy (PF-4 : before 49.6 ± 29.8, after 24.0 ± 19.6ng/ml, β-TG : before 117.5 ± 64.0, after 64.6 ± 47.1ng/ml;paired t-test, p < 0.0001, respectively). These results suggest that saffron is effective in treatment of the symptoms of “Oketsu” in allergy and collagen disease patients. Moreover, PF-4 and β-TG may be useful biomarkers of the degree of “Oketsu.”

A Case of Heparin Induced Thrombocytopenia with Circulating Anti-PF4/Heparin Antibody

Heparin-induced thrombocytopenia (HIT) is usually caused by anti-platelet factor 4 (PF4)/heparin antibodies, leading to intravascular platelet activation. Circulating anti-PF4/heaprin antibody (IgG, IgA and IgM) was detected by ELISA (Asserachrom HPIA, Diagnostia Stago, Asnieres, France) in a 61-yearold man with coronary artery disease and dyspnea on exercise. He had undergone a coronoary angiography using 2,000 unit of heparin before a procedure. On admission, laboratory testing revealed a platelet count of 296x10(9)/L and aPTT (activated partial thromboplastin time) of 38.1 sec. The fall in the platelet count was progressive, resulting in 42% and 53% of platelet counts on the 6th and 12th days after intravenous heparin administration, respectively. He was discharged after coronary artery bypass graft. On discharge, platelet count was normalized to be 212x10(9)/L. On the 7th day after dischage, anti-PF4/heaprin antibody was detected by ELISA (Asserachrom HPIA, Diagnostia Stago, Asnieres, France).

Heparin-Induced Thrombocytopenia during Continuous Veno-Venous Hemodiafiltration after Total Aortic Arch Replacement: A case report / 대한마취과학회지

Heparin-induced thrombocytopenia (HIT) is an immunologically mediated complication of heparin therapy resulting in the consumption of platelets and a catastrophic thromboembolism. Both the clinical and laboratory features are important for a diagnosis of HIT. There have been 3 case reports of suspected HIT in Korea. However none have satisfied the laboratory features. We experienced a case of HIT, which satisfied both clinical and laboratory features, in a patient who received heparin during continuous veno-venous hemodiafiltration (CVVHD) used to treat acute renal failure that developed after a total aortic arch replacement with a cardiopulmonary bypass. The decreased platelet count and obstruction of extracorporeal filter of CVVHD by the blood clot was observed while receiving unfractionated heparin. The serum from the patient contained the anti heparin-platelet factor 4 antibody, and the condition was thus diagnosed as HIT. Argatroban, which is a direct thrombin inhibitor, was used to treat the thrombosis.

Effects of piretanide on plasma fibrinolytic activity, platelet aggregation and platelet factor-4 release in man.

Piretanide, 4-phenoxy-3-(pyrrolidinyl)-5-sulphamoyl benzoic acid, apart from being an efficient diuretic, enhances endogenous plasma fibrinolytic activity after a single dose of 6 mg administered by oral route. After ingestion of the drug, acceleration of fibrinolytic acitivity became manifest within 1 h, reached its peak in 3 h and was associated with a fall in fibrinogen and diminished urokinase excretion. Piretanide did not cause lysis of fibrin in vitro. Primary platelet aggregation, induced by adenosine-diphosphate, was inhibited by piretanide. In in vitro experiments piretanide led to effective inhibition of adenosine-diphosphate-induced platelet aggregation with complete inhibition at 5 mM concentration. Piretanide led to a highly significant decrease of platelet factor-4 release.

INFLUENCE OF ENDOSTATIN AND PF-4 ON LYMPHANGIOGENESIS / 解剖学报

Objective Try to discover the safe and effective inhibitors of lymphangiogenesis. Methods Lymphatic endotheial cells from pig thoracic duct were isolated and cultured. The observation of specimens by LM and EM was made. The control group, endostatin and PF-4 experimental groups were set. Methods of the scraped line and MTT were used to examine their inhibitive effect on the lymphangiogenesis. Results The P value is