0.05);in treating gradeⅢto IV aGVHD,the effectiveness of FK506 with MP was significantly better than that of CsA with MP(P

Objective To assess the value of three-dimensional contrast-enhanced ultrasonography (3D-CEUS) in evaluation of hepatic arteries variants.Methods Both two-dimensional contrast-enhanced ultrasonography (2D-CEUS) and 3D-CEUS were used to assess 30 patients including living donor candidates and patients with upper abdomen tumors.The reference standard was operation or CTA or DSA or MRA,and the accuracy for detecting hepatic artery variants provided by the two methods was evaluated.Arterial anatomic types were defined by using Michels classification.Results The total accuracy for detecting hepatic artery anatomy types by 2D-CEUS was 40.0% (12/30),while 83.3% (25/30) by 2D-CEUS.For convention anatomy types the accuracy on 2D-CEUS and 3D-CEUS were 40.9%(9/22)and 90.9%(20/22),respectively.The difference was statistically significant (P<0.05).For anatomy variants types the accuracy on 2D-CEUS and 3D-CEUS were 37.5%(3/8)and 62.5%(5/8),respectively.No significant difference between these two methods was observed.Conclusion 3D-CEUS was a new method in diagnosis of hepatic arteries anatomy types with practical clinical value in evaluation of the living liver donors.

Objective To investigate the effects of ZMS,the active component of Zhimu,on amyloid precursor protein(APP) and ?-site APP cleaving enzyme 1(BACE1) in HEK293sw cells. Methods HEK293sw cells were pretreated with different concentrations of ZMS in routine culture medium for 24 h,and then serum-free medium with ZMS was used for further culture for 48 h.Cells were examined for expression of APP and ?-secretase BACE1 with Western blotting and RT-PCR,and were then examined for BACE1 activity with fluorescence method. Results 1 ?mol/L and 10 ?mol/L ZMS significantly decreased the expression of BACE1 mRNA and protein,while had no effect on the expression of APP mRNA and protein.It was indicated by fluorescence analysis that 10 ?mol/L ZMS significantly decreased the ?-secretase activity.Conclusion ZMS significantly decreases the activity and expression of ?-secretase BACE1,while has no effect on the expression of APP in HEK293sw cells.

Objective To explore the changes of humoral immunity in neonates with hypoxic-ischemic encephalopathy(HIE)and observe the influence of perinatal factors on humoral immune function.Methods Sixty-two neonates with HIE were enrolled and 30 healthy neonates were selected as control group.The percentages of CD19,CD25 of B lymphocyte were detected by Flow Cytometr.The levels of IgG,IgM,IgA and complement C3,C4 were detected by immune rate nephewlometry.The results were analyzed by SPSS 11.5 software.Results 1.The percentages of CD19+(17.93?3.10)%,CD19+CD25+(0.64?0.42)% and the levels of IgM(0.13?0.05)g/L,IgA(0.14?0.07)g/L and complement C3(0.62?0.12)g/L,C4(0.10?0.03)g/L in neonates with HIE were significantly lower than those in the healthy term neonates(Pa0.05).2.There were dignificant diffe-rences of the percentages of CD19+,CD19+CD25+ and the levels of IgM,IgA,complement C3,C4 among the different stages of HIE(Pa

OBJECTIVETo detect the expression of surface molecule CD96 on stem cell (LSC) in children with acute leukemia, and to explore its clinical significance.

METHODSBone marrow mononuclear cells were isolated in 69 children with newly diagnosed acute leukemia. CD34(+)CD38(-)CD123(+) LSCs were separated from these cells by flow cytometry (FCM) and then cultured, and CD96 expression on LSCs was detected by FCM. R-banding technique was used to analyze the karyotypes of the 69 children, and the data of their routine blood and immunological tests were collected.

RESULTSCD96 was mainly expressed in children with acute myelogenous leukemia, and expressed to a lesser extent in those with acute lymphoblastic leukemia (P<0.05). The median expression level of CD96 in Uyghur children was 23.4%, versus 21.2% in Han children (P>0.05). The majority of children with CD96-positive children presented poor-prognosis karyotypes. Compared with CD96-negative children, children with CD96-positive children had a significantly lower complete remission rate (P<0.05) and significantly higher infection and relapse rates after chemotherapy (P<0.05).

CONCLUSIONSChildren with acute leukemia who have CD96-positive LSCs have a poor prognosis. CD96 may be a new indicator of prognosis in children with acute leukemia.

Adolescent ; Antigens, CD ; analysis ; Child ; Child, Preschool ; Humans ; Infant ; Leukemia, Myeloid, Acute ; drug therapy ; genetics ; immunology ; pathology ; Neoplastic Stem Cells ; chemistry ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; drug therapy ; genetics ; immunology ; pathology ; Prognosis

AIMTo analyse the impurities of gatifloxacin.

METHODSThe impurity of gatifloxacin were analysized and determinated by RP-HPLC/electrospray ionization mass spectrometry with a Zorbax SB-C18(4.6 mm x 150 mm ID, 5 microns). The mobile phase was 3% acetic acid/acetonitrile-3% acetic acid/water (15:85). The two compounds were synthesized: 1-cyclopropyl-6-fluoro-1, 4-dihydro-8-methoxy-7-(1-piperazinyl)-4-oxo-3-quinolinecarboxylic acid (DMP) and 1-cyclopropyl-6-fluoro-1, 4-dihydro-8-hydro-7-(3-methy-1-piperazinyl)-4-oxo-3-quinolinecarboxylic acid (DMO). Their liquid chromatogram, UV, MS were compared with those of the impurity of gatifloxacin.

RESULTSThe mass of the impurity was 14 less than that of gatifloxacin. It means the impurity was CH2 less than gatifloxacin. The tR (HPLC), UV and MS of DMP were the same as those of the impurity of gatifloxacin.

CONCLUSIONBased on the tR (HPLC), UV and MS, the impurity of gatifloxacin is confirmed as DMP.

Anti-Infective Agents ; analysis ; chemistry ; Chromatography, High Pressure Liquid ; Drug Contamination ; Fluoroquinolones ; analysis ; chemistry ; isolation & purification ; Molecular Structure ; Spectrometry, Mass, Electrospray Ionization

OBJECTIVETo investigate the effect of p65 gene inhibited by siRNA on neuronic differentiation in the marrow mesenchymal stem cells (MSCs).

METHODSThe MSCs were transfected with Rn-p65-siRNA. Fasudil hydrochloride induced MSCs differentiating into neurons. The non-transfected group and negative control group (transfected with negative control siRNA marked by Cy3) were used as controls. The fluorescence expressed by transfected MSCs were observed under inverted fluorescence microscope at 24 h,48 h and 72 h after transfected with negative control siRNA. The viability of MSCs was detected by MTT at 24 h, 48 h and 72 h after transfected with Rn-p65-siRNA. The expressions of p65 mRNA and protein in MSCs were detected by RT-PCR and Western blot respectively. The expressions of p65 protein, NSE, MAP-2 and glial fibrillary acidic protein (GFAP) were detected by immunocytochemical method after transfection for 6 h.

RESULTSThe fluorescence of MSCs was mostly displayed after transfection of 72 hours and the efficiency of transfection was up to 83.3% ± 3.8%. Meanwhile, the p65 mRNA and p65 protein expressed by MSCs of transfected group were significantly decreased (P < 0.05); MTT displayed that the viability of MSCs was also significantly reduced (P < 0.05). The best efficiency of induction was observed in the transfected group. There were higher expressions of NSE and MAP-2 than the other group (P < 0.05).

CONCLUSIONThe p65 gene inhibited by siRNA can promote the marrow mesenchymal stem cells to differentiate into neurons.

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine ; analogs & derivatives ; Animals ; Cell Differentiation ; Glial Fibrillary Acidic Protein ; metabolism ; Mesenchymal Stromal Cells ; cytology ; Neurons ; cytology ; RNA, Messenger ; RNA, Small Interfering ; Rats ; Transcription Factor RelA ; antagonists & inhibitors ; metabolism ; Transfection

The study was purposed to investigate the differentiation ability of mesenchymal stem cells (MSCs) into myocardial cells in vitro. Rat bone marrow-derived MSCs were labeled and co-cultured with neonatal rat cardiomyocytes (CM) for 5 - 7 days. The expression of cell surface antigens was detected by flow cytometry, and the expression of muscle-specific marker myosin and troponin T in labeled cells was detected by immunofluorescence. The results showed that in vitro cultured MSCs expressed CD90, CD44, CD105, CD54, not expressed CD34, CD45, CD31. After co-cultured with neonatal rat CM, labeled MSCs differentiated into cardiomyocyte-like cells expressing myosin and troponin T. It is concluded that MSCs can differentiate into cardiomyocyte-like cells when co-cultured with neonatal myocardial cells in vitro. In co-culture of two kind of cells in ratio of four to one showed obvious efficacy differentiating MSCs into CMs.
Animals ; Bone Marrow Cells ; cytology ; Cell Differentiation ; Cells, Cultured ; Coculture Techniques ; Mesenchymal Stromal Cells ; cytology ; Myocytes, Cardiac ; cytology ; Rats ; Rats, Wistar

In order to analyze the incidence and high-risk factors of invasive fungal infections among recipients of allogeneic hematopoietic stem cell transplantation (allo-HSCT), 180 cases of allo-HSCT were enrolled in this study. The incidence and risk factors of IFI were analyzed by method of Kaplan-Meier and Cox regression model. The results showed that an incidence of IFI in 35 cases (19.5%) were detected, with 1 case proven and 34 cases probably diagnosed, which was composed of 18 cases (51.4%) of aspergillosis and 17 cases (48.6%) of candidosis. There was significant difference in one-year overall survival rate between patients with (34.3%) or without (53.8%) IFI. In univariate analysis, risk factors of IFI included: pretransplant fungal infection or colonization, unrelated donor (peripheral blood or bone marrow stem cell) transplantation, acute GVHD, extensive chronic GVHD and the use of methylprednisolone. In multi-variate analysis, the following risk factors of IFI were found:unrelated donor for allogeneic peripheral blood or bone marrow stem cell transplantation, acute GVHD and pretransplant fungal infection or colonization acute GVHD (RR: 2.399, 1.589, and 0.836). It is concluded that IFI is a frequent complication and one of the leading causes of mortality among recipients of allo-HSCT. As for patients with higher risk of IFI, early interventions should be taken.
Adolescent ; Adult ; Aspergillosis ; epidemiology ; etiology ; Candidiasis ; epidemiology ; etiology ; Child ; Child, Preschool ; China ; epidemiology ; Female ; Graft vs Host Disease ; complications ; Hematologic Neoplasms ; therapy ; Hematopoietic Stem Cell Transplantation ; adverse effects ; Humans ; Incidence ; Male ; Middle Aged ; Risk Factors ; Young Adult

This study was purposed to investigate the effect of prostaglandin E2 (PGE2) on proliferation of peripheral blood T lymphocytes, and to evaluate the regulatory role of PGE2 on immunological balance between Th1/Th2 and Tc1/Tc2 lymphocytes. The peripheral blood mononuclear cells (PBMNC) were stimulated by anti-human CD3 monoclonal antibody (mAb) and anti-human CD28 mAb, and were cultured in the presence of different concentration of PGE2 for 120 hours. The proliferation of peripheral blood T lymphocytes was assayed according to the manufacture protocol of BrdU Kit; the IFN-gamma and IL-4 levels in supernatants cultured for 24, 48, 72 and 120 hours were detected by ELISA; the ratios of CD4+IL-4+ T cells/CD4+ IFN-gamma+ T cells and CD8+IL-4+ T cell/CD8+IFN-gamma+ T cells were determined by flow cytometry. The cells cultured without PGE2 were used as control. The results indicated that (1) with the raising of concentration of PGE2, the inhibitory rate of T cell proliferation in vitro significantly increased (p=0.001). There was significant positive correlation between inhibitory rate of T cells and PGE2 concentration (correlation coefficient=0.889, p=0.000). (2) the difference between the IFN-gamma concentrations in supernatant cultured for 120 and 72 hours in test groups had no statistical significance (p=0.917). The IFN-gamma concentration increased continually with prolonging of culture time in control group (p=0.046). The IFN-gamma concentrations produced at different times in test group were significantly lower compared with those in control group (p<0.05). The IL-4 concentrations produced at different time had no significant change in test groups (p=0.400). The IL-4 concentration in 24 hours in control group was significantly higher than that at 48, 72 and 120 hours in control group (p=0.007, 0.003 and 0.002). After cultured for 24 hours the IL-4 concentration in test group was significantly lower than that in control group (p=0.037), but after cultured for 48, 72 and 120 hours, the IL-4 concentration in test group did not show statistical difference in comparison with control group (p>0.05). (3) the proportions of CD4+IFN-gamma+T cells in test group and in control group had no significant difference (p=0.767). The proportion of CD4+IL-4+T cells in test group was slightly higher than that in control group (p=0.051). The ratio of CD4+IL-4+T cells to CD4+IFN-gamma+ T cells in test group was significantly higher than that in control group (p=0.011). The proportions of CD8+IFN-gamma+ T cells in test group and in control group had no statistical difference (p=0.441). The proportion of CD8+IL-4+T cells in test group was significantly higher than that in control group (p=0.015). The ratio of CD8+IL-4+ T cells to CD8+IFN-gamma+ T cells in test group were obviously higher than that in control group(p=0.038). It is concluded that the PGE2 inhibits the proliferation of T lymphocytes in vitro. PGE2 influences the production of IFN-gamma and IL-4, and significantly influences peak appearance of IFN-gamma produced by T lymphocyte. PGE2 can continuously inhibit the production of IFN-gamma, but its continuous effect on IL-4 is no significant. PGE2 enhances the ratio of CD4+IL-4+T lymphocytes to CD4+IFN-gamma+T lymphocytes and the ratio of CD8+IL-4+T lymphocytes to CD8+IFN-gamma+T lymphocytes, and regulates development of T cells toward Th2/Tc2 cells.
Cell Proliferation ; drug effects ; Dinoprostone ; pharmacology ; Flow Cytometry ; Humans ; Lymphocyte Activation ; drug effects ; Lymphocyte Count ; T-Lymphocytes, Cytotoxic ; drug effects ; immunology ; Th1 Cells ; drug effects ; immunology ; Th2 Cells ; drug effects ; immunology