0 5).Conclusions The measurement of platelet membrane GPⅡbⅢa complexes by FCM can provide a simple, sensitive and reliable method for G T diagnosis,and also can be applied to analysis of GT family pedigree.

BACKGROUND: There still was not any report about inducing stem cells into matured cells to form products in vitro.OBJECTIVE: To induce CD34+ cells of umbilical cord blood to differentiate into mature megakaryocytes, and to investigate the mechanism of production of platelets.DESIGN, TIME AND SETTING: This cytology in vitro study was conducted at the Central Laboratory of Xiangya Hospital and Xiangya Third Hospital from 2004 to 2006. MATERIALS: Umbilical cord was collected from healthy full-term pregnant puerperants at the Xiangya Hospital.METHODS: The CD34+ cells were isolated from umbilical cord blood by magnetic activated cell sorting (MACS) and then cultured in 24-well culture plate at 5x107/L in StemPro-34 serum-free medium, supplemented with L-glutamine, saturated human transferrin, CaCl2, insulin, deionized bovine serum albumin and recombinant human thrombopoietin at 37℃, under 0.05 volume fraction CO2 saturated humidity to be differentiated into megakaryocytes for 14-21 days. Cell medium was absorbed, and centrifuged to obtain supernatant. Samples were centrifuged again, and then supernatant was removed. The remaining was platelet-like particles in cell culture plate. Platelet was isolated from normal platelet-rich plasma.MAIN OUTCOME MEASURES: The following parameters were measured: morphological changes in cultured cells and platelet-like particles in supematant; results of immunohistochemistry; observation results under a microscope; platelet aggregation; CD41 expression.RESULTS: At day 10, silk-like substances were found in megakaryocyte culture medium, with the presence of platelet-sized particles. The production of platelet-sized particles reached a peal at day 16. Cultured cells were strongly positively for platelet-specific antigen GP Ⅱb Ⅲa. Under the optical microscope, mature megakaryocytes were detected, with the presence of some immature megakaryocytes, and platelet-sized particles were found surrounding megakaryocytes. Under the electron microscope, a majority of mature megakaryocytes and a few apoptotic megakaryocytes were detected, and platelet-sized particles in the supernatant had the same size and structure with the platelet in the platelet-rich plasma. Some platelet surfaces were smooth or irregular. Platelet-sized particles in the supematant aggregated in response to thrombin as platelets in normal platelet-rich plasma. Flow cytometry demonstrated that the cultured platelets had the same high expression rate of CD41 as the platelets from platelet rich plasma.CONCLUSION: Umbilical cord blood CD34+ cells can be induced to differentiate into pudfied and mature megakaryocytes and platelets in vitro.

Objective To explore the anatomic characteristics of the peroneal perforator branches and its clinical application as vascularized flap transfer. Methods Twenty fresh cadaver specimen with 40 sides lower limbs were used in this study.Lead oxide gelatin was injected to the whole body,lower extremity radiaograph, spiral CT scan was then used to construct three demention visual model. The peroneal artery and its perforators were dissected,number of peferators,distance to fibular head,diameter and the length of the vascular pedicles were measured and analyzed. From July 2005 to October 2009, forty-three cases with skin defects were performed vascularized transfer in our study,surviving rate and postoperative function were followed up for 6 months to 2 years.Results Perforators were seen most at (9.80 ± 0.93)cm,(13.40 ±0.90) cm,(17.20 ± 1.13)cm,and (21.30 ± 0.77)cm beneath the fibular head with the artery branch diarneter(1.33 ± 0.39) mm,(1.30 ± 0.46)mm,(1.17 ± 0.30)mm,and (1.22 ± 0.23)mm,respectively,while the pedicle length was (5.87 ± 0.73)cm,(5.83 ± 1.73)cm,(5.44 ± 1.09)cm,and (5.10 ± 1.93) cm respectively.In clinic,42/43 free flaps survived.Postoperative outlook were satisfied except in 7 cases,the flaps looked bulky and needed secondary revision.All the donor calves showed good apperaence and function.Conclusion There are 4 regular perferators in lateral calf, while perforators in the middle 1/3 are bigger with relatively longer vascular pedicles which are appropriate for vascularized transfer.

Objective To prepare the water soluble chrysin-hydroxypropyl-β-cyclodextrins inclusion compound and widen the administration path of chrysin .Methods The cogrinding method had been used to prepare chrysin-hydroxypropyl-β-cyclodextrins in-clusion compound .The PXRD, DSC and IR techniques had been used to characterize the inclusion compound .Results Chrysin and hydroxypropyl-β-cyclodextrins had formed the inclusion compound , and the formation of the inclusion compound could increase solubili-ty by 120.7 times.Conclusion The inclusion compound preparation method was simple and available , which was suitable to improve the bioavailability .

This study examined the sodium nitroprusside (SNP) toxicity to central nervous system (CNS) in treatment of patients with aortic dissection (AD). The medical records of 191 AD patients who were admitted to Tongji Hospital, China, from Jan. 1998 to Feb. 2009 were retrospectively analyzed. There were 140 cases of hypertension (73.3%) and 13 cases of Marfan syndrome (6.8%) among the 191 AD patients. A total of 157 patients were given SNP treatment. The toxic reactions of CNS occurred in 18 subjects (11.5%). Most of the adverse reactions occurred on the fifth day following SNP injection. SNP infusion rate was significantly higher in patients who developed CNS toxicity. It was suggested that systemic hypertension is the most common predisposing factor for AD. The combination of SNP with a β-receptor blocker is a medical therapy commonly used in patients with AD. Cyanide and thiocyanate toxicity from SNP treatment is always the consequence of prolonged drug infusion or relatively high dose administration.

OBJECTIVE: To determine the expression of DLK1 gene in acute leukemias (AL) and its function in erythroid differentiation of K562 cells. METHODS: We detected the expression of DLK1 gene in 65 different acute leukemia categories (a test group) and 34 normal bone marrow controls (a control group) with RT-PCR. DLK1 protein in 20 out of the 65 AL patients and 13 of the 34 controls was assayed by Western blot. The K562 cell line was induced to erythroid differentiation by hemin. We observed the relationship between its expression and erythroid differentiation. RESULTS: Both leukemia cells and normal marrow cells expressed DLK1. The expression of DLK1 mRNA in patients in the test group was higher than that in the control group (P=0.018), while there was no significance between acute lymphoblastic leukemia and acute myelogenous leukemia (P>0.05).The expression of DLK1 mRNA in the test group at onset had no relation with the WBC and platelet count in the total peripheral blood, and the same was true for blast cell rates in bone marrow cells.The level of DLK1 protein in the test group was higher than that in the control group, which was consistent with the mRNA expression (P=0.042). The expression of DLK1 mRNA decreased gradually with K562 cells towards hemin-induced erythroid differentiation. CONCLUSION DLK1 gene may be involved in leukemia,but the mRNA level of DLK1 has no relation with some clinical characteristics of AL patients at onset. DLK1 may inhibit the erythroid differentiation of K562 cells.
Acute Disease ; Adolescent ; Adult ; Aged ; Calcium-Binding Proteins ; Case-Control Studies ; Cell Differentiation ; drug effects ; genetics ; Cell Transformation, Neoplastic ; genetics ; Child ; Child, Preschool ; Erythroid Cells ; pathology ; Erythroid Precursor Cells ; pathology ; Female ; Humans ; Intercellular Signaling Peptides and Proteins ; genetics ; metabolism ; K562 Cells ; Leukemia ; genetics ; metabolism ; Male ; Membrane Proteins ; genetics ; metabolism ; Middle Aged ; RNA, Messenger ; genetics ; metabolism ; Young Adult

This study examined the sodium nitroprusside (SNP) toxicity to central nervous system (CNS) in treatment of patients with aortic dissection (AD). The medical records of 191 AD patients who were admitted to Tongji Hospital, China, from Jan. 1998 to Feb. 2009 were retrospectively analyzed. There were 140 cases of hypertension (73.3%) and 13 cases of Marfan syndrome (6.8%) among the 191 AD patients. A total of 157 patients were given SNP treatment. The toxic reactions of CNS occurred in 18 subjects (11.5%). Most of the adverse reactions occurred on the fifth day following SNP injection. SNP infusion rate was significantly higher in patients who developed CNS toxicity. It was suggested that systemic hypertension is the most common predisposing factor for AD. The combination of SNP with a β-receptor blocker is a medical therapy commonly used in patients with AD. Cyanide and thiocyanate toxicity from SNP treatment is always the consequence of prolonged drug infusion or relatively high dose administration.
Adult ; Aged ; Aged, 80 and over ; Aorta ; surgery ; Central Nervous System ; drug effects ; Female ; Humans ; Male ; Middle Aged ; Nitroprusside ; adverse effects ; therapeutic use ; Young Adult

Objective The recurrence of perimitral atrial tachycardia is common after initial ablation of persistent atrial fibrillation (AF). The aim of the study is to explore a preferable ablation approach for perimitral atrial tachycardia in the redo ablation of persistent AF. Methods Seventy-four patients with perimitral atrial tachycardia after initial ablation for persistent AF were included in our study. Patients were distinguished into either the group of having ablation during tachycardia (Group A) or the group having ablation after cardioversion to sinus rhythm (Group B) according to the different ablation strategies. The procedural endpoints were pulmonary vein isolation and bidirectional conduction block of all the ablated lines. The primary endpoint of the study was freedom from atrial tachyarrhythmia recurrence during the follow-up period. Results There were statistical differences in baseline clinical data between the 2 groups. During the redo procedure, conduction recovery rate across the mitral isthmus (MI),cavotricuspid isthmus and left atial roofline were 100％, 40.5％ and 48.6％ respectively. The procedural time, fluoroscopy time, mapping time were longer in the patients of group A. During a mean follow-up of (16.9±6.3) months, 31 (72.1％) patients in group A and 21(67.7％) patients in group B maintained in sinus rhythm in the absence of antiarrhythmic durgs (P =0.771) . Conclusion In patients with perimitral atrial tachycardia after initial ablation for persistent AF,ablation in sinus rhythm is a more simplified method and as effective as ablation during tachycardia.

Objective To identify the electrophysiological charateristics and cause of ridge gap related reentry after MI ablation in atrial ifbrillation patients. Methods Activation and entrainment mapping was performed in 82 redo cases for OAT recurrence in whom MI was ablated during the index produre. Once ridge gap related reentry was conifrmed, detailed mapping was performed in MI and ridge region. In addition, in 36 cases undergoing MI ablation and fulfilling criterion for bidirectional block, differential pacing was repeated at the ridge to identify a ridge gap. Results Out of 82 redo cases for OAT recurrence in whom MI was ablated during the index produre, 7 (8.5%) was found to be ridge gap related reentry. TCL was (247.9±19.2) ms, and the left atrial endocardial activation time was (145.4±17.7) ms, accounting for (58.5±3.2)%of TCL. However, wide double potential was recorded along the previous ablated MI line where PPI was (34.3±6.6) ms longer than TCL, while PPI was signiifcantly shorter at the ridge[PPI-TCL (11.4±3.9) ms, P<0.001]. Tachycardia was terminated at the ridge in 6 cases and at the corresponding site in coronary sinus in 1 case. No recurrence was found during follow-up for (11.1±4.5) months. In addition, in 36 patients undergoing MI ablation in whom criterion of bi-directional block was fuliflled, conduction gap located at the ridge was found in 5 (13.9%) cases. Conclusions MI ridge gap related reentry is a distinctive OAT, in which the ridge was used as the critical isthmus, whereas the previous ablated MI line is not part of the reentry. MI pseudo-block due to the ridge gap may lead to this type of recurrent tachycardia.

Objective Cryoballoon ablation of pulmonary vein (PV) ostia often induces a vagal response.This prospective study was designed to assess the effectiveness of prophylactic intravenous administration of atropine on hemodynamic impairment induced by cryoballoon ablation in patients with atrial fibrillation.Methods Twenty-five patients with paroxysmal atrial fibrillation undergoing cryoballoon ablation were prospectively enrolled and assigned to either the trial group on the control group.First twelve patients (the trial group) were administered 1 mg of atropine before deflation of the cryoballoon,while the following 13 patients (the control group) were given atropine only after the onset of the hemodynamic variation (decrease in heart rate and/or blood pressure).Treatment was considered effective when the hemodynamic variations were restored.Results In the trial group,three patients with transient hypotension did not require further supportive care throughout the procedures and one patient with hypotension required supportive management.In the control group,hypotension,bradycardia and mixed bradycardia with hypotension requiring supportive care occurred in six,three,and three patients,respectively.Overall,the rate of marked vagal responses was significantly lower when prophylactic atropine was administrated (4/12 vs.12/13 patients,P ＜ 0.01).Conclusions Atropine is effective in the prevention of all types of vasovagal responses induced by cryoballoon ablation in patients with atrial fibrillation.