In the light of requirements of medical equipment simulation system, the overall framework of joint control system of medical equipment integrated support in the theater is designed in this paper. Such modules are developed with Delphi as communication module, module for equipment basic information, module for equipment video information and module for medical faculty information. A joint control system suiting integrated support simulation sand table for medical equipment is complteted,which provides a multidimensional informational platform for integrated demonstration of medical equipment.

Objective To investigate the expression of peroxisome proliferators-activated receptor ? (PPAR?) in trophoblast and relation between PPAR? ligands and trophoblast invasion.Methods We examined the expression of PPAR? by immunohistochemistry,immunocytochemistry and real time quantitative PCR.We next examined,using the cytotrophoblast culture model,the biological role of PPAR? ligands in vitro.Results PPAR? was mainly localized in the nuclei of villous cytotrophoblast and extravillous cytotrophoblast of cell islands and cell columns.In villous tissue and cultured trophoblast from early first trimester,the level of expression of PPAR? mRNA and protein was 36.0?5.1,13.4?3.1 and 1.35?0.08,1.13?0.11;from late first trimester it was 23.3?5.5,6.1?1.3 and 1.17?0.03,0.86 ?0.05,and the expression of PPAR? was obviously decreased (P

Objective To construct a novel enhanced green fluorescent protein (EGFP) and glial cell line-derived neurotrophic factor (GDNF) recombinant baculovirus. Methods The target gene(EGFP and GDNF) was cloned into baculovirus transfer vector pFastBacDual, pFB-EGFP-GDNF was constructed and restriction enzyme analysis was conducted. pFB-EGFP-GDNF was transposited with baculovirus shuttle vector (Bacmid) into DH10Bac competent cells, and recombination baculovirus vector Bacmid-EGFP-GDNF was constructed. The plasmid was extracted and PCR was performed for identification. Bacmid-EGFP-GDNF was transfected with Sf9 insect cell package virus by liposomal transfection method. Immunofluorescent staining was employed to detect the expression of EGFP and GDNF protein in St9 cells. Results The target gene fragment was correctly cloned into pFastBaeDual vector, and recombinant Bacmid was constructed. Bacmid-EGFP-GDNF was successfully transfected, and higher virus titer was obtained. The coexpression of GDNF and EGFP protein in Sf9 cells was identified by immunofluorescent staining. Conclusion The recombinant baculovirus Bacmid-EGFP-GDNF can be successfully constructed, and the protein of EGFP and GDNF is coexpressed in St9 cells, which paves a way for the research of GDNF gene therapy.

OBJECTIVETo investigate the cell composition in granulocyte colony-stimulating factor One (rhG-CSF) primed bone marrow grafts (G-BM) from donors with different characteristics.

METHODShundred and fifty healthy donors were injected subcutaneously rhG-CSF (5 microg x kg(-1) x d(-1)) in five consecutive days. Bone marrow and peripheral blood stem cells were harvested at day 4 and day 5. The number of CD3, CD3+ CD4+, CD3+ CD8+, CD14+ , CD34+ and CD3+ CD4- CD8- cells were determined by multicolor flow cytometry. Cell composition of G-BM from donors with different characteristics, including sex, age, weight, pregnancy were analysed.

RESULTSThe absolute number of nuclear cells (NCs), lymphocytes, CD3+, CD3+ CD4+, CD3+ CD8+, CD14+, CD34+ and CD3+ CD4- CD8- cells in G-BM were 31 050 (12 200 -58 100), 2122 (506 - 6618), 1344 (307 - 4791), 692 (145 - 3038), 616 (112 - 2575), 986 (265 -2958), 63 (11 -505) and 83 (9 -390) per microliter, respectively. The number of NCs [33 800 (18 600 - 57 100)], CD34+ cells [76 (22 -505)], and CD3+ CD4+ CD8- regulatory T cells [97 (11 - 380)] harvested from younger donors (aged < or = 38 years) were significantly higher than those from older ones (aged > 38) [28000 (12200-58100), 49 (11-220), and 65 (9 - 389) per microliter (P = 0.027, < 0.001 and = 0.001, respectively)]. Compared with that in donors with peripheral blood monocytes (PBMs) < or = 0.37 x 10(9)/L, higher number of NCs in G-BM [27 150 (13 800 - 58 100) vs 33 550 (12 200 - 57 100), P = 0.005] were collected in donors with PBMs > 0.37 x 10(9)/L. Multivariate analysis showed that donors age (< or = 38 vs > 38 years; P = 0.01) and monocyte number in peripheral blood (< or = 0.37 x 10(9)/L vs > 0.37 x 10(9)/L; P = 0. 003) were factors predicting for NC yields, and donors age ( < or = 38 vs > 38 years; P < 0.001) was factor predicting for yields of CD34+ cells (P < 0.001) and CD3+ CD4- CD8- regulatory T cells (P = 0.001) collection.

CONCLUSIONDonor age is a factor for the yields of NCs, CD34+ cells, and CD3+ CD4- CD8- regulatory T cells collection in G-BM, and donor's PBMs more than 0.37 x 10(9)/L, is another factor affecting NCs harvests.

Adolescent ; Adult ; Age Factors ; Antigens, CD34 ; Bone Marrow Cells ; immunology ; Female ; Granulocyte Colony-Stimulating Factor ; therapeutic use ; Hematopoietic Stem Cell Mobilization ; Humans ; Male ; Middle Aged ; Recombinant Proteins ; T-Lymphocyte Subsets ; immunology ; Tissue Donors ; Young Adult

OBJECTIVETo investigate the optimal time for second allogeneic peripheral blood stem cell grafts (PBSC) harvest from healthy donors after in vivo recombinant human granulocyte colony-stimulating factor application (rhG-CSF).

METHODSThirty-eight healthy donors of second collection (group A) were treated with subcutaneous rhG-CSF \[5 microgxkg(-1)xd(-1)\] for five consecutive days and followed by leukapheresis on day 5 and 6. The control group (group B) was thirty-eight healthy donors who had received a first PBSC collection previously. Group A was reclassified as group C (< or = 9 months) and group D (> 9 months) according to the 75% quantile of interim time between first and second collection. The quantities of lymphocytes of CD3(+), CD3(+)CD4(+), CD3(+)CD8(+), CD14(+), CD34(+) cells and CD3(+)CD4(-)CD8(-) T cells were determined by multi-color flow cytometry.

RESULTSThe median number of CD3(+)CD8(+) (25.51 x 10(8)) and CD34(+) cells (0.51 x 10(8)) in group A were significantly lower than that (31.55 x 10(8) and 0.70 x 10(8) respectively) in group B (P < 0.05), and so did the CD3(+)CD8(+) (23.42 x 10(8)) and CD34(+) cells (0.42 x 10(8)) in group C than that in group B (P < 0.05). There was no statistical difference in median numbers of T cell subsets, monocytes, and CD34(+) cells between group B and group D (P > 0.05). The cell ratios of CD4(+)/CD8(+), CD14(+)/CD3(+) and CD3(+)CD4(-)CD8(-) T/CD3(+) in PBSC in group A, group C, and group D were similar to that in group B (P > 0.05). Sperman analysis showed a positive correlation between the total CD34(+) cells in second collection and the interval time from first to second collection (r = 0.357, P = 0.028).

CONCLUSIONNine months after the first collection maybe an optimal time for the second PBSC collection. For those who undergo second PBSC collection within 9 months, more circulation blood should be extracted to ensure enough immunological and hematopoietic compositions.

Adolescent ; Adult ; Cytapheresis ; methods ; Female ; Granulocyte Colony-Stimulating Factor ; therapeutic use ; Hematopoietic Stem Cell Mobilization ; Humans ; Male ; Middle Aged ; Peripheral Blood Stem Cell Transplantation ; Recombinant Proteins ; Time Factors ; Tissue Donors ; Transplantation, Homologous ; Young Adult

The study was purposed to investigate the effect of bone marrow graft collection on the composition of peripheral blood stem cell grafts in healthy donors after recombinant human granulocyte colony-stimulating factor (rhG-CSF) application in vivo. Sixty-two healthy donors were treated with rhG-CSF [5 microg/(kg.d)] injected subcutaneously for five consecutive days. Donors were divided into groups A and B, and 31 donors were there in each group. Bone marrow grafts and peripheral blood stem cell grafts were harvested on the day 4 and 5 respectively in group A. In group B, only peripheral blood grafts were collected on both the day 4 and 5. The quantities of the cell components, CD3(+), CD3(+)CD4(+), CD3(+)CD8(+), CD14(+), CD34(+) cells and CD3(+)CD4(-)CD8(-) T cells were determined by multi-color flow cytometry. The results showed that median counts of nuclear cells (1.56 x 10(5)), lymphocytes (8.56 x 10(4)), CD3(+) (6.12 x 10(4)), CD3(+)CD4(+) (3.38 x 10(4)), CD3(+)CD8(+) (2.27 x 10(4)), CD14(+) (3.83 x 10(4)), CD34(+) cells (744) and CD3(+)CD4(-)CD8(-) T cells (3588) per microliter of peripheral blood stem cell grafts in group A were similar to counts of nuclear cells (1.40 x 10(4)), lymphocytes (7.34 x 10(4)), CD3(+) (5.32 x 10(4)), CD3(+)CD4(+) (3.06 x 10(+)), CD3(+)CD8(+) (1.83 x 10(4)), CD14(+) (3.21 x 10(4)), CD34(+) cells (554) and CD3(+)CD4(-)CD8(-) T cells (3120) in group B (p > 0.05). There were no difference in the ratios of CD4(+) cells/CD8(+) cells, CD14(+) cells/CD3(+) cells and CD3(+)CD4(-)CD8(-) T cells/CD3(+) cells in peripheral blood stem cell grafts between group A [1.52 (0.54 - 2.87)], [0.57 (0.15 - 1.64)], [0.064 (0.018 - 0.673)] and group B [1.68 (0.31 - 3.35)], [0.59 (0.18 - 1.25)], [0.063 (0.021 - 0.136)] (p > 0.05). It is concluded that no effect of bone marrow graft collection on the composition of peripheral blood stem cell grafts in the same donor is found after rhG-CSF application in vivo, bone marrow grafts and peripheral blood stem cell grafts can be collected respectively or simultaneously.
Adolescent ; Adult ; Bone Marrow Cells ; cytology ; Bone Marrow Transplantation ; Female ; Granulocyte Colony-Stimulating Factor ; therapeutic use ; Hematopoietic Stem Cell Mobilization ; Humans ; Male ; Middle Aged ; Peripheral Blood Stem Cell Transplantation ; Recombinant Proteins ; Tissue Donors ; Young Adult

The study was aimed to analyze the difference of naive and memory CD4(+) and CD8(+) T-cell subsets between steady-state bone marrow (SS-BM) and recombinant human granulocyte colony-stimulating factor (rhG-CSF) mobilized peripheral blood grafts (G-PB). Four CD4(+) and CD8(+) T-cell subsets classified according to the expression of CD45RA and CD62L were determined by three-color flow cytometry. The results showed that the percentage of CD4(+), CD8(+) T-cell subsets and the ratios of CD4/CD8 in G-PB were significantly higher than those in SS-BM (p < 0.05). The percentage of CD4(+) naive T-cells in G-PB was significantly lower than that in SS-BM (p < 0.001). As compared with SS-BM, the percentage of CD4(+) effector memory T-cells was significantly high in G-PB (p < 0.001). There were no significant differences in the percentages of the four CD8(+) T-cell subsets between SS-BM and G-PB (p > 0.05). The percentage of CD4(+)CD62L(+) T-cells in G-PB was significantly lower than that in SS-BM (p = 0.001). The absolute numbers of CD4(+) and CD8(+) T-cell subsets, the eight naive and memory CD4(+) and CD8(+) T-cell subsets were significantly higher in G-PB than those in SS-BM (p < 0.001). It is concluded that the difference of naive and memory CD4(+) and CD8(+) subsets between G-PB and SS-BM may partially explain why the incidence and severity of acute graft-versus-host disease (GVHD) was similar and the incidence of chronic GVHD was different after transplantation with SS-BM or G-PB.
Adolescent ; Adult ; Aged ; Bone Marrow Transplantation ; CD4-CD8 Ratio ; CD4-Positive T-Lymphocytes ; immunology ; CD8-Positive T-Lymphocytes ; immunology ; Female ; Graft vs Host Disease ; etiology ; Granulocyte Colony-Stimulating Factor ; therapeutic use ; Hematopoietic Stem Cell Mobilization ; Humans ; Male ; Middle Aged ; Peripheral Blood Stem Cell Transplantation ; Recombinant Proteins ; Young Adult

OBJECTIVETo investigate the regular pattern of Cytomegalovirus (CMV)-specific T cells (CTL) immune reconstitution after human leukocyte antigen (HLA) matched sibling donor allogeneic bone marrow(BM) plus peripheral blood hematopoietic stem cell (PBSC) transplantation.

METHODSCTL from seventeen patients after transplantation was detected by flow cytometry, the IFN-γ secretion ability of CTL by enzyme-linked immunospot (ELISPOT) assay, and clonal analysis of TCR Vβ subfamily by gene scan assays. The relationship between CTL reconstitution and CMV infection was studied.

RESULTSBoth number and function of recipients CTL reached to normal control level at 30 d post-transplantation. The recipients achieved a high frequency CTL with IFN-γ response and restoration of T-cell receptor β (TCR Vβ) repertoire at one year post-transplantation. CTL with the central memory CD45RO(+)CD62L(+) cell phenotype expanded in PB when CMV was reactivated. The incidence of CMV reactivation was 35.83% (17.91% - 63.10%) after transplantation, and none of them developed CMV disease.

CONCLUSIONAfter HLA matched related donor transplantation using mixed grafts, immune recovery to CMV seems to be early and fast. The incidence of CMV infection and disease are low.

Adult ; Cytomegalovirus ; immunology ; Female ; HLA Antigens ; immunology ; Hematologic Diseases ; immunology ; surgery ; Hematopoietic Stem Cell Transplantation ; Humans ; Male ; Middle Aged ; Peripheral Blood Stem Cell Transplantation ; Siblings ; T-Lymphocytes, Cytotoxic ; immunology ; Tissue Donors ; Young Adult

Objective To clarify the seasonal and geographical difference and pathogen patterns so as to provide reference of prevention and control of the disease through analyzing the epidemical characteristics of reported hand-foot-mouth disease(HFMD)cases in China,2008-2009.Methods We analysed the epidemiological data of HFMD from the Chinese national notifiable infectious diseases reporting system in 2008 and 2009.Results There were 488 955 and 1155525 reported HFMD cases in 2008 and 2009,in China,of which 1165 and 13810 were severe and 126 and 353 were fatal,respectively.The notification rates were 50.09/10 million,68.47/10 million and 59.04/10 million in high,medium and low latitudes areas,respectively.The epidemic periods in Medium and Low latitude were from 12 to 24 weeks in 2009,and in high latitude it was from 23 to 35 weeks.HFMD cases were concentrated mainly in 5 year-old or even younger children,accounted for 92.23% of the total cases.The incidence rate of two years old appeared to be the highest.The features of severe and death case concentrated in lower age groups were more evident,and the proportion of severe case and case fatality rate under 1 vears old was higher than that in other age groups.We also noticed that with the increasing of age,the proportion of severe case and case fatality rate had a decreasing trend.There was a difference between the pathogens seen.The relative risk(RR)for an human enterovirus 71(HEV71)isolate was higher among severe case than in common cases(RR=1.82),whereas the RR for an EV71 isolate was higher among the death cases than in common cases (RR=2.11).There was seasonal variation of pathogen composition.There were 477 clusters of cases from 2008 to 2009,of which 389 found in preschools,47 in rural villages,outbreaks of clusters were mainly from April to July. Conclusion The HFMD epidemics was increasing in 2009.The epidemic of HFMD in different latitudes area and seasons was different.Children of five year old or under were the major population groups at risk,of HFMD.The younger ones had higher risk of becoming severe and death cases.HEV71 and coxsackievirus A16 were both the major etiologic agents of HFMD.The preschool and rural villages were the main senings of clusters of cases.

This study was purposed to investigate the relation of monocyte counts in peripheral blood (PB) at the first collection of allograft to the amount of CD34(+) cells in the mixture of recombinant human granulocyte colony-stimulating factor (rhG-CSF)-primed bone marrow graft (G-BM) and rhG-CSF mobilized peripheral stem cell grafts (G-PB). 70 healthy donors were treated with rhG-CSF [5 microg/(kg.d)] injected subcutaneously for 5 consecutive days. Bone marrow grafts and peripheral blood grafts were harvested on the 4th and 5th days respectively. Blood cell counts at the first collection of allografts were determined by blood analyzer XL2100, the amount of CD34(+) cells in G-BM and G-PB were determined by flow cytometry. The results showed that the monocyte counts in the PB of the 70 healthy donors were (1.15 +/- 0.60) x 10(9)/L. The amount of total CD34(+) cells in the G-BM, G-PB and mixture grafts were (5.85 +/- 2.93) x 10(7), (1.33 +/- 0.77) x 10(8), and (1.92 +/- 0.86) x 10(8) respectively. Pearson and Spearman correlation analysis indicated that the counts of monocyte at the first collection of allografts correlated positively with the amount of total CD34(+) cells in the G-BM (correlation coefficient, r = 0.265, p = 0.027), G-PB (r = 0.340, p = 0.004) and mixture grafts (r = 0.398, p = 0.001). Stepwise Linear Regression Model analysis also showed that the counts of monocytes in the PB correlated positively with the amount of CD34(+) cells in the G-BM, G-PB and mixture grafts (p = 0.027, 0.004 and 0.001 respectively). The sensitivity and specificity of the monocyte counts to predict the amount of CD34(+) cells in the mixture grafts were 71% and 70% respectively (p = 0.007). In conclusion, the monocyte counts in the PB at the first collection of allografts after rhG-CSF treatment of healthy donors may be a simple and practical indicator for harvest of CD34(+) cell amount in the mixture grafts.
Adolescent ; Adult ; Antigens, CD34 ; immunology ; Blood Cell Count ; Bone Marrow Transplantation ; Female ; Flow Cytometry ; Granulocyte Colony-Stimulating Factor ; Hematopoietic Stem Cell Mobilization ; Humans ; Male ; Middle Aged ; Monocytes ; cytology ; immunology ; Peripheral Blood Stem Cell Transplantation ; Recombinant Proteins ; Tissue Donors ; Young Adult