Objective To analyze the influence of different factors and their relating correlation results on platelet transfusion during the bone marrow empty window period on the patients who have undergone allogeneic peripheral blood stem cell transplantation (allo-HSCT) with retrospective analysis of case-control data.Methods Clinical data of 153 cases were collected by the clinical blood management and evaluation information system with discharge diagnosis of allo-HSCT in the hematology department of The First Affiliated Hospital of Nanchang University within a time frame from January 2014 to December 2016.A total of 90 cases were considered valid for retrospective analysis according to the case exclusion criteria.The average transfusion dose for patients with allo-HSCT during the bone marrow empty window period was defined as the threshold value which divided the 90 cases into the observation group of 38 cases receiving more than 6 Units of platelet transfusion and the control group of 52 cases with less than 6Units of platelet transfusion.The amount of platelets transfused during the bone marrow empty window period,clinical indexes include Hb,ANC,Plt,SF before pretreatment,platelet engraftment time and the number of mononuclear cells implanted were compared and analyzed by Logistic regression.Results (1) There was no significant difference between the two groups in gender,age,primary diagnosis,HLA matching,Hb before pretreatment and the number of mononuclear cells implanted (P＞0.05).The ANC(×109/L) (1.24±0.57 vs 3.36±1.33) and Plt(×109/L) (43.55±68.29 vs 126.62±84.73) counts before pretreatment in the observation group were significantly lower than those in the control group(P＜0.05).SF(μg/L) (2351.05 ± 1 587.96 vs 1 000.96± 362.97)before pretreatment and P LT recovery time (d) (16.84± 2.47 vs 12.73 ± 1.65)was significantly higher than that in the control group(P＜0.05).Donor-recipient ABO blood group typing incompatibility (15 vs 10) was significantly higher than the control group (P＜0.05);(2) Single factor Logistic regression analysis showed that ABO blood group matching,clinical indexes include ANC,Plt,SF before pretreatment,PLT recovery time were statistically significant,Only ANC before pretreatment and PLT recovery time had significant effect on the platelet transfusion during bone marrow empty window period in patients with allo-HSCT in multivariate Logistic regression analysis(P＜0.05).Condusion The ANC before pretreatment and PLT recovery time are independent factors for platelet transfusion of the bone marrow empty window period in patients with allo-HSCT.The PLT recovery time is an independent risk factor,which indicates that the longer the duration of PLT implantation,the greater the amount of platelet transfusion will be needed.Besides,the ANC before pretreatment is the independent protective factor,which indicates that the greater the ANC,the smaller the amount of platelet transfusion is required.

This study examined the effect of cholic acid (CA) on cultured cardiac myocytes (CMs) from neonatal rats with an attempt to explore the possible mechanism of sudden fetal death in intrahepatic cholestasis of pregnancy (ICP). Inverted microscopy was performed to detect the impact of CA on the beating rates of rat CMs. MTT method was used to study the effect of CA on the viability of CMs. CMs cultured in vitro were incubated with 10 μmol/L Ca(2+)-sensitive fluorescence indicator fluo-3/AM. The fluorescence signals of free calcium induced by CA were measured under a laser scanning confocal microscope. The results showed that CA decreased the beating rates of the CMs in a dose-dependent manner. CA could suppress the activities of CMs in a time- and dose-dependent manner. CA increased the concentration of intracellular free calcium in a dose-dependent manner. Our study suggested that CA could inhibit the activity of CMs by causing calcium overload, thereby leading to the sudden fetal death in ICP.

Objective To observe the effect of Kangnaoye on angiogene-sis in rats after focal cerebral ischemia reperfusion.Methods The male SD rats were randomly divided into 6 groups: sham operation group , model group, Kangnaoye high, middle and low dosages groups (24,12,6 g· kg -1· d-1), and nimodipine group(1 mg· kg -1· d -1).The mod-els of cerebral ischemia reperfusion were established in rats by methods of middle cerebral artery occlusion ( MCAO).Immunohistochemical method was used to observe the change of the microvessel density ( MVD ) after cerebral ischemic and the expression of vascular endothelial growth ( VEG), HGF and CD31 protein of rats which were killed at the 0.5, 3, and 7 d of reperfusion injury respectively 2 h after cerebral ischemia.The nervous function deficit scores were evaluated at the 2 , 24 , and 48 h af-ter reperfusion.Results Compared with model group , the neurological behavior performance in Nimodipine and Kangnaoye treatment groups were significantly improved , with an increased number of MVD , and en-hanced vascular endothelial growth factor ( VEGF) and hepatocyte growth factor( HGF) protein levels ( P<0.05 or P<0.01 ) , especially in the Kangnaoye middle dosage group.Conclusion Kangnaoye has neuropro-tective effect against focal cerebral ischemic injury by improving the ex-pression of VEGF and HGF , which is one of possible anti -ischemic mechanisms.

OBJECTIVETo analyse the clinical outcome and the prognostic factor of childhood acute myeloid leukemia (AML).

METHODSDisease-free survival (DFS), event-free survival (EFS) and overall survival (OS) rates were estimated by Kaplan-Meier method and prognostic factors were evaluated by Cox regression with SPSS in 141 childhood AML in our hospital from August 1995 to July 2004. The patients were divided into 2 groups: acute promyelocytic leukemia (APL) as group A and AML other than APL as group B.

RESULTSOf the 90 group B patients, 54.4% (49/90) achieved complete remission (CR) after one course chemotherapy , with a total CR rate of 76.7%. The cumulative 5 year DFS and OS rate for group B patients were (28.4 +/- 9.0)% and (35.5 +/- 6.3)%, the 51 group A patients were (94.3 +/- 4.0)% and (81.4 +/- 5.7)%, and for total 141 AML patients were (56.9 +/- 6.3)% and (53.3 +/- 4.8)% respectively. Multivariate analysis demonstrated that higher bone marrow blast cell percentage at diagnosis, CR after more than one course of chemotherapy and less than six courses of consolidation chemotherapy were risk prognostic factors in childhood AML other than APL (P < 0.05).

CONCLUSIONThe prognosis of childhood APL is better, while of childhood t(8;21) AML is no better than other FAB subtypes.

Adolescent ; Bone Marrow Cells ; cytology ; Child ; Child, Preschool ; Disease-Free Survival ; Female ; Humans ; Infant ; Leukemia, Myeloid, Acute ; mortality ; therapy ; Male ; Prognosis ; Regression Analysis ; Retrospective Studies ; Treatment Outcome

To evaluate the impact of trisomy 8 on cytobiological and clinical features of acute myelomonocytic and monocytic leukemia (M(4), M(5)), a total of 56 cases of acute myelomonocytic and monocytic leukemia were investigated. Karyotypes were analyzed by G-banding or R-banding. The immunotypes in all cases were detected by flow cytometry. And the clinical characteristics at the first visit were analyzed retrospectively. The results showed that thirty-four of 56 (60.7%) patients had normal cytogenetics; 10 (17.9%) patients had trisomy 8 in their karyotypes, including 3 (5.4%) patients with trisomy 8 as the sole aberration; and 12 (21.4%) patents had other cytogenetic abnormalities (except trisomy 8). All trisomy 8 cases demonstrated a increased expression frequency of surface markers of myeloid progenitor cells CD34 (P < 0.01) and CD117 (P < 0.05) and a decreased expression frequency of surface markers of mature monocytes CD11c (P < 0.01) and CD14 (P < 0.05), compared with normal cytogenetics cases. Patients with trisomy 8 were slightly older (P < 0.05), which had lower percentages of peripheral blasts (P < 0.05) and lower WBC (P < 0.05) than the patients without trisomy 8. Patients with trisomy 8 had a shorter disease-free survival time than that of patients with normal cytogenetics (P < 0.05). It is concluded that trisomy 8 may play an important role in the pathogenesis and progression of acute myelomonocytic/monocytic leukemia (M(4)/M(5)), whic seems to be related with a block in differentiation of monocytes. Therefore, trisomy 8 may be an adverse prognostic factor for patients with M(4) or M(5).
Adolescent ; Adult ; Aged ; Antigens, CD34 ; analysis ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Bone Marrow Transplantation ; CD13 Antigens ; analysis ; Chromosome Banding ; Chromosomes, Human, Pair 8 ; genetics ; Female ; Flow Cytometry ; HLA-DR Antigens ; analysis ; Humans ; Immunophenotyping ; Karyotyping ; Leukemia, Monocytic, Acute ; genetics ; immunology ; therapy ; Leukemia, Myelomonocytic, Acute ; genetics ; immunology ; therapy ; Male ; Middle Aged ; Peripheral Blood Stem Cell Transplantation ; Prognosis ; Trisomy

Objective To study the thermal insulation properties of vacuum insulated panel(VIP)used as materials for blood transportation kits.Methods A VIP transportation kit was taken as test group,and a conventional transportation kit was taken as control.Phase change cool storage materials of the same amount and 4℃water bags were put into both kits.A recordable electronic thermometer was used to record the temperature within the two kits,and free hemoglobin(FHb)and potassium ion concentration were measured at 0, 4, and 7 days after blood storage.Results The temperature in the conventional transportation kit increased faster after 36 h, and was significantly higher than in the VIP transportation kit until 60 h.Besides,the VIP transportation kit kept the temperature under 10℃ for(60.7 ±0.6)h, compared to (54.2 ±3.0)h in the conventional transportation kit.FHb concentration was significantly lower in the VIP transportation kit[(605.26 ±74.63)mg/L]than in the conventional transportation kit[(1327.60 ±187.41)mg/L]at day 7(d 7),so was the potassium ion concentration in the VIP transportation kit[(22.7 ±0.4)mmol/L]compared to[(24.6 ±0.6) mmol/L]in the transportation kit at d 7.Conclusion The VIP transportation kit keeps the temperature under 10℃ for a longer time,and the blood quality of preservation is better than that of the conventional transportation kit.Novel heat preservation material can improve health support ability,and is of great value and significance.

The aim was to study the apoptotic induction effect of thapsigargin on leukemia cell line K562 and its possible mechanism. After the treatment of leukemia cell line K562 by thapsigargin, morphological change of apoptotic cells was investigated by AO/EB fluorescent staining under fluorescent microscope; apoptosis rate was determined with annexin V-FITC/PI double staining by flow cytometry; intracellular calcium concentrations ([Ca(2+)]i) were measured by fluorescence spectrophotometer with calcium sensitive fluorescence indicator Fura-2/AM; mitochondrial transmembrance potentials (Delta Psi m) was detected on flow cytometry through staining of Rhodamine (Rh123); the changes of caspase-3, -7, -9, -12, cytochrome C, GRP78 proteins were detected by Western blot. The results showed that K562 cells cultured in 4 micromol/L thapsigargin for 48 hours exhibited typical morphological changes of apoptotic cells under fluorescent microscope, including shrinkage of cell, condensation of chromatin, breakage of nuclear, formation of apoptotic bodies, fluorescence of yellow green and pellet observed in early apoptoyic cells and hyacinth fluorescence of chromatin showed in late apoptotic cells. 24 and 48 hours after exposure to 1, 2, 4, 8 micromol/L thapsigargin, the apoptotic rates of K562 were respectively 7.51%, 11.65%, 23.22%, 30.56% and 12.85%, 20.27%, 31.51%, 44.16%, in dose-dependent manner, and were statistically significant when compared with the controls (P < 0.05). The apoptotic rate of K562 was dose- and time-dependent in experiment range. The enhancement of [Ca(2+)]i and the decrease of the Delta Psi m in K562 cells were induced by thapsigargin and were dose-dependent in experiment range, compared with control, P < 0.05. Western blot results indicated that cleavage and activation of caspase-3, -7, -9, -12, releasing of cytochrome C from mitochondria, upregulation of GRP78 expression at the endoplasmic reticulum were induced in K562 cells after 24 hours exposure of 4 micromol/L thapsigargin. It is concluded that thapsigargin induces endoplasmic reticulum stress-induced apoptosis in K562 cells. Endoplasmic reticulum is a novel important initiatory site of apoptosis in cells; the cleavage and activation of caspase-3, -7, -9, -12 play very important role in endoplasmic reticulum stress-induced apoptosis of K562 cells and is one of the important mechanisms for thapsigargin-induced apoptosis. Thapsigargin-induced apoptosis in K562 cells is associated closely with the disruption of the Delta Psi m and the release of cytochrome C from mitochondria, mitochondria participates in endoplasmic reticulum stress-induced apoptosis in K562 cells.
Apoptosis ; drug effects ; Calcium-Transporting ATPases ; antagonists & inhibitors ; Caspase 3 ; metabolism ; Caspase 7 ; metabolism ; Cytochromes c ; metabolism ; Endoplasmic Reticulum ; drug effects ; enzymology ; Enzyme Inhibitors ; pharmacology ; Heat-Shock Proteins ; metabolism ; Humans ; K562 Cells ; Leukemia ; pathology ; Mitochondria ; drug effects ; Molecular Chaperones ; metabolism ; Thapsigargin ; pharmacology

Acute monocytic leukemia is a distinct subtype of acute myeloid leukemia (AML) with characteristic biology and clinical features. This study was designed to compare the immunophenotypical features and clinical manifestations of the patients with AML-M(5a) to that of patients with AML-M(5b), and to identify differences between M(5a) and M(5b) and to explore their relations. A total of 58 cases of de novo adult patients with AML M(5) were investigated. Immunofluorescence analysis by flow cytometry was performed to determine the immunophenotype of the leukemic cells in all cases. Meanwhile, clinical data of these cases were studied retrospectively. The results showed that the immunophenotypes of monocytic leukemic cells in patients with AML M(5) were heterogeneous, and CD68 and CD11b were expressed higher in patients with AML M(5a), compared with that in patients with AML M(5b) (P < 0.01). The significant differences in sex, extramedullary infiltration, WBC counts of peripheral blood, complete remission rate and disease-free survival (DFS > 300 days) between the patients with AML M(5a) and M(5b) did not exist (P > 0.05). It is concluded that the special individual immunophenotype features can be detected in patients with either of AML M(5a) or M(5b), and that expressions of CD68 and CD11b were much higher in M(5a). It seems that the complete remission rate and disease-free survival of patients with M(5a) and M(5b) are not different from that of currently available therapy.
Adolescent ; Adult ; Antigens, CD ; analysis ; Antigens, Differentiation, Myelomonocytic ; analysis ; CD11b Antigen ; analysis ; Female ; Fluorescent Antibody Technique ; Humans ; Immunophenotyping ; Leukemia, Monocytic, Acute ; classification ; immunology ; Male ; Middle Aged ; Prognosis

This study was done for investigating the frequency and proliferative feature of mesenchymal stem/progenitor cells (MSPC) in human umbilical cord blood (CB) and for searching a new seed cell for tissue engineering. Mononuclear cells was separated by Ficoll-Hypaque from cord blood and suspended in DMEM culture medium supplemented by 2% fetal bovine serum. The adherent CB cells were cultured and expanded at same medium. The results showed that the frequency of CB-MSPC was 0.5 x 10(-6) [(0.2 - 0.8) x 10(-6)]. The CB-MSPC showed a fibroblast-like morphology and retained their morphological feature at least after 20 sub-passages, and could extensively be expanded by about 1.3 x 10(7) times as much. The conclusion is that low serum DMEM culture could maintain the proliferation and differentiation potential of CB-MSPC. CB-MSPC might be a favorable seed cell for tissue engineering and regeneration.
Cell Adhesion ; Cell Division ; Fetal Blood ; cytology ; Humans ; Mesenchymal Stromal Cells ; physiology ; Tissue Engineering

To investigate the value of apoptosis of the allo-antigen specific T cells induced by Fas/FasL pathway in order to prevent GVHD in allo-transplant, the CD34(+) cells were transfected with FasL or not, used as effector cells, mixed with allo-antigen specific T lymphocytes with presence or absence of IFN-gamma or IL-2. After 5 days, apoptosis of T cells was detected by TdT nick end mediated dUTP labeling (TUNEL) and flow cytometry. The effects of IFN-gamma or IL-2 on apoptosis of CD34(+) cells of graft induced by Fas/FasL pathway observed as controls. The apoptosis incidence of T cells was (12.1 +/- 1.5)% when CD34(+) cells transfected with FasL were used as effector cells, that was much higher than that T cells with CD34(+) cells non-transfected (p < 0.01). In the presence of IFN-gamma or IL-2, apoptosis incidence reached to (20.1 +/- 2.3)% or (17.6 +/- 1.3)% respectively (p < 0.01). When sFasL was added to CD34(+) cells freshly isolated or induced with IFN-gamma or IL-2, the incidence or apoptosis of CD34(+) cells was (7.8 +/- 0.8)%, (18.7 +/- 1.6)% (p < 0.01) or (7.9 +/- 1.0)% (P > 0.05) respectively. The results suggest that it is possible to induce apoptosis of the allo-antigen specific T cells in grafts activated by allo-antigen by exogenous Fas ligand expressed on receptor cells and that may hopefully provide a new method to prevent GVHD
Animals ; Antigens, CD34 ; analysis ; Apoptosis ; Cell Communication ; physiology ; DNA, Complementary ; genetics ; Fas Ligand Protein ; Humans ; Interferon-gamma ; pharmacology ; Membrane Glycoproteins ; genetics ; physiology ; Mice ; T-Lymphocytes ; cytology ; physiology ; Transfection ; fas Receptor ; biosynthesis