OBJECTIVE:To observe clinical effects of alprostadil combined with nitric oxide in the treatment of congenital heart disease complicated with pulmonary hypertension and its effects on immune function. METHODS:In retrospective study,40 cases of congenital heart disease complicated with pulmonary hypertension in thoracic and cardiovascular surgery department of our hospital during Aug. 2009 to Feb. 2015 were selected and divided into nitric oxide group(single group)and alprostadil+nitric oxide group(combination group)according to therapy method,with 20 cases in each group. Both groups received conventional surgery. Single group was given extrinsic inhalation of nitric oxide(volume fraction of 20×10-6)after surgery;combination group was addi-tionally given intravenous pump of alprostadil 5 ng/(kg·min) for consecutive 72 h,on the basis of combination group. The heart rate,left ventricular ejection fraction(LVEF),mean arterial pressure,pulmonary arterial pressure,T lymphocyte and immunoglob-ulin(Ig)of children before and after treatment,and the occurrence of ADR were compared between 2 groups. RESULTS:Before treatment,there was no statistical significance in above indexes between 2 groups (P>0.05). After treatment,heart rate,LVEF, mean arterial pressure,the levels of CD3+,IgA,IgM and IgG in 2 groups had no significant change,but CD4+,CD4+/CD8+,Treg/CD4+ were increased significantly;pulmonary arterial pressure,the levels of CD8+,Th1,Th2,Th1/Th2 and Th17/CD4+,supple-ment C3 and supplement C4 were decreased significantly,compared to before treatment. Above indexes in the combination group was significantly better than the single group,with statistical significance (P<0.05). No obvious ADR was found in 2 groups. CONCLUSIONS:Alprostadil combined with nitric oxide could significantly reduce the pulmonary hypertension in children with congenital heart disease complicated with pulmonary hypertension,improve the level of T lymphocytes,and decrease the levels of supplement C3 and supplement C4 with good safety.

BACKGROUND:Some studies have demonstrated that red blood celllysate added into the bone marrow can increase the efficiency of isolating and purifying mesenchymal stem cells, in order to obtain high-purity and high-quantity bone marrow mesenchymal stem cells.
OBJECTIVE:To isolate and culture bone marrow mesenchymal stem cells of rabbit with red blood celllysis in vitro for exploring proliferation characteristics and performing the biological identification of cells.
METHODS:Bone marrow suspension was col ected by puncturing the tibia with medul o-puncture needle. Red blood celllysis was added to the bone marrow suspension, and bone marrow mesenchymal stem cells were isolated and cultured. Numbers of primary cells were recorded at 4, 7, 10, and 13 days later And Growth curves of the cells at passages 2-5 were drawn for comparison of proliferative characteristics. Inverted phase contrast microscope was used to observe the morphological changes of cells. CD34, CD44 antigens of bone marrow mesenchymal stem cells were identified by flow cytometry and immunity fluorescence.
RESULTS AND CONCLUSION:The adherent bone marrow mesenchymal stem cells mainly showed s spindle shape, with homogeneous nuclei, prominent nucleoli, and rich cytoplasm, which were positive for CD44 antigen and negative for CD34 antigen. The primary cells exhibited an“S”shape. Passage 4 cells had a better proliferative ability, rapider growth and more counting of cells as compared with other generations. These findings indicate that red blood celllysis method is a feasible ways of culturing bone marrow mesenchymal stem cells 0 in vitro, and passage 4 cells have the strongest proliferation capacity.