BACKGROUND:Erythropoietin and progenitor cel transplantation both have therapeutic effects on lower extremity arterial occlusive disease.
OBJECTIVE:To investigate the erythropoietin modification effect and magnetic resonance imaging feasibility of superparamagnetic iron oxide (SPIO)-labeled endothelial progenitor cel s in vitro.
METHODS:Rat bone marrow-derived endothelial progenitor cel s at logarithmic growth phase were randomized into four groups:endothelial progenitor cel group, SPIO labeled transfection group (pcDNA3-EPO transfection fol owed by SPIO labeling), SPIO labeled empty vector group (empty plasmid transfection fol owed by SPIO labeling), and SPIO labeling group (only SPIO labeling). 4.7T MRI was used to observe SPIO-labeled endothelial progenitor cel s. Cel proliferation, cel cycle distribution, and expression of erythropoietin protein in the four groups were measured.
RESULTS AND CONCLUSION:MRI findings showed with the increasing cel number, gradual y lowered signal intensity on T1-weighted imaging (T1WI), T2WI and T2*WI was seen, and the reduction in the signal intensity was the maximum on T2*WI sequence and the minimum on T1WI sequence. For T1WI, T2WI and T2*WI sequences, the minimum number of cel s was 2×104, 1×104 and 0.5×104, respectively. Cel proliferation and cel cycle distribution showed no significant difference among three SPIO labeling groups. In addition, the expression of erythropoietin protein was only found in the SPIO-labeled transfection group. These findings showed that under SPIO labeling, erythropoietin gene-modified endothelial progenitor cel s show no changes in cel proliferation and cel cycle, and the 4.7T MR is capable of imaging SPIO-labeled erythropoietin gene-modified endothelial progenitor cel s in vitro.

Objective To investigate the care measures of microspheres arterial embolization in the treatment of primary liver cancer after TACE.Methods The chemotherapy drugs were injected through the catheter superselectively inserted to the left and right hepatic arteries,microspheres and super liquefied iodipin were used for tumor embolization,preoperative and postoperative care were provided to patients.Results Through the full nursing care of 50 cases of liver cancer patients,the occurrence of adverse effects and complications after surgery were reduced,enthusiasm of patients with treatment were confirmed,and the effect of the treatment was improved.Conclusions Treatment of liver cancer by hepatic artery perfusion chemo-embolization is an effective means,using scientific methods of effective care for patients is very important.

Objective To explore the transfection efficiency of primary lymphocytes from human peripheral blood by different methods to acquire the method with higher transfection efficiency.Methods Mononuclear cells from human peripheral blood were isolated using Ficoll-Hypaque.Cell viability was detected by Trypan blue staining.Suspending lymphocytes were sucked out and were incubated in 24-well plate after cultured in 6-well plate for 2 h.Activated lymphocytes were transfected by electroporation with plasmid(PEGFP-N1).Resting or activated lymphocytes were transfected by lentivirus vector(LVGFP) single infection or repeated infection,respectively.Green fluorescence protein (GFP) was detected under the fluorescence microscopy and percentage of positive cells was checked by flow cytometry at different time points after infection.At the same time,the effectiveness of lentivirus infection was compared under different conditions.Results Purity of mononuclear cells isolated by Ficoll-Hypaque was 95 ％ and its viability was over 95 ％.The percentage of lymphocytes obtained with a uniform shape was 90 ％-95 ％.Scattered fluorescence was observed by electroporation under the conditions of voltage 2 100 V,pulse width 10 ms,pulse number 1 for lymphocyte,while fluorescent became weaker over time and no green fluorescent was observed after transfection for 72 h.After resting lymphocytes were infected once for 48 h by lentivirus vector,green fluorescent was not found and positive cells were less than 1％.1％-5 ％ of activated lymphocytes could express GFP after single lentivirus infection and the expression levels were enhanced with concentration increasing,while 5 ％-10 ％ of activated lymphocytes showed strong green fluorescent by repeated lentivirus infection.In contrast with electroporation,the fluorescent with lentivirus infection was stronger over time.Conclusion Repeated lentivirus infection could efficiently transfect exogenous genes into activated lymphocytes for stable expression.

AIM: It is found that some active priciples of ginseng can enhance and activate body immune system, and have many bioactivities such as anti-tumor, anti-aging and anti-radiation. This study examined the effect of ginsenoside-Rh2 (GS-Rh2) on proliferation and apoptosis in human myeloid leukemia cell strain HL60, and analyzed the dose- and time-dependent manners of GS-Rh2. METHODS: Experiments were performed at the Department of Clinical Laboratory of Jilin Medical College from July to August in 2006. ①Rh2 was purchased from Hongjiu Biotech Co., Ltd. (batch number 050801), and prepared into 50 g/L stock solution by dissolving in pH 7.4 phosphate buffer saline. The HL60 cell strain was purchased from Shanghai Institute of Cell Biology of Chinese Academy of Sciences. ②HL60 cells in logarithmic growth phase were inoculated into 3?108 L-1 cell suspension. After the cells were cultured for 6 hours, 100 ?L GS-Rh2 at different concentrations (5,10,20,40,80 mg/L) was added respectively. After the cells were administrated for 48 hours, cell inhibition ratio (IR) was evaluated by MTT colorimetric assay. The 50% inhibitory concentration (IC50) was worked out. HL60 cell was acted with this concentration for different time (6, 12, 24, 48 and 72 hours), cell inhibition ratio (IR) at different time points was evaluated by MTT colorimetric assay, and compare it to the control. After the IC50 of GS-Rh2 acted for 48 hours, HL60 cells were observed with an inverted microscope. HL60 cell was stained by Giemsa, and the typical apoptosis cells were discovered. RESULTS: ①Dose-effect relationship: When the concentration of GS-Rh2 was 5,10,20,40 mg/L, the IR of GS-Rh2 to the growth of HL60 cells was increased gradually in obviously dose-dependent manner. The IR was similar between 80 mg/L and 40 mg/L. After the cells were administrated for 48 hours, the IC50 value was 13.0 mg/L. ②Time-effect relationship: After the concentration of IC50 of GS-Rh2 (13.0 mg/L) acting on HL60 line at different time points (6, 12, 24, 48 and 72 hours), cell IR was increased gradually (F=9.32,P

Objective To investigate whether the B vitamins supplements would lower total homocysteine and reduce the risk of recurrent stroke in patients with recent ischemic stroke.Methods A prospective,open,case-controlled clinical trial.One thousand ischcmic stroke patients with hyperhomocysteinemia were followed up.They were assigned to receive either a daily dose of B vitamins (folic acid 2.5 mg,B6 25 mg,B12 500 μg,treatment group,n =500) or not ( control group,n =500) for a period of 2 years.Total homocysteine level,demographic information and traditional risk factors were collected as well as recurrent cerebral infarction were noted.Results Homocysteine levels were significantly reduced in the active treatment group,reduction of total homocysteine was 14.7％ at 3 months and 19.2％ at 24 months ( F =94.39,P ＜0.05 ).The risk of ischemic recurrent stroke with clinical sign within 2 years was 13.6％ for the active treatment group and 14.0％ for the control group ( risk ratio =0.99,95％ CI 0.68—1.42).The risk of recurrent stroke with only MRI or CT brain scan evidence was 4.8％ for both groups (risk ratio =1.11,95％ CI 0.62—2.02 ).The risk of total recurrent stroke was 18.4％ for the treatment group and 18.8％ for the control group ( risk ratio =0.96 ; 95％ CI 0.73—1.26 ),but these effects were not significant.Conclusion A significant benefit of secondary prevention with long-term reductions in blood homocysteine levels with B vitamins supplementationin during the 2 years of follow-up is not yet proven.

Objective To evaluate the clinical effect of therapy of the trisacryl gelatin microspheres combinee the gelatin sponge particle on embolize the bronchial artery in acute massive hemoptic patients. Methods One huneree cases with massive hemoptysis were selectee as our subjects ane eivieee into control ane research group(n = 50 for each group). Patients in control group were given only gelatin sponge particle,ane in research group were given the trisacryl gelatin microspheres combinee the gelatin sponge particle to embolize the bronchial artery. All cases were followee up for more than 12 months. Ane the effect of therapy was recoreee. Results In research group,42 cases(84. 0% ,42 / 50)were got the bleeeing stop immeeiately after embolization,7 cases in 72 h(14. 0% ,7 / 50),ane the effective rate of hemostasis was 98. 0%(49 / 50). In the control group,41 case(82. 0% ,41 / 50)were got the stop bleeeing immeeiately,8 cases in 72 h(16. 0% , 8 / 50),ane the effective rate of hemostasis was 98. 0%(49 / 50). There was no statistic eifference between two groups(P > 0. 05). After more than one year follow-up,3 cases(6. 12% )were reoccurree in the therapy group ane 15 cases(30. 61% )was in the control group. The eifference was significant between two groups after surgery for one year( χ2 = 9. 801,P < 0. 01 ). There was no serious complication in patients of two groups. Conclusion The operation of BAE is effective therapy for the massive hemoptoe,ane it is provee to be a safe,effective ane lower rate of recurrence approach of the trisacryl gelatin microspheres combinee the gelatin sponge particle for eouble embolzation the bronchial artery.

Objective To investigate the effects of B lymphocyte-induced maturation protein-1 ( Blimp-1) on the number and function of splenic lymphocytes.Methods The mice with defective Blimp-1 in T cells were generated by cross-breeding B6.Blimp-1flox/flox mice with B6.Lck-Cre mice.The mononuclear lymphocytes isolated from spleen of T cell conditional Blimp-1 knockout (Blimp-1CKO) mice and wild type ( WT) C57/B6 mice were comparatively analyzed.Alterations of CD4+T and CD8+T cell subsets, the secre-tion of cytokines as well as the expression of C-C chemokine receptor type 7 ( CCR7 ) and Sphingosine-1-phosphate receptor 1 (S1P1) in mice from the two groups were analyzed by flow cytometry.The changes of CD19+B cell subsets were also detected.Results Compared with WT mice, the total numbers of mononu-clear cells, T and B lymphocytes were all significantly increased in Blimp-1CKO mice ( P<0.05) .The ab-solute numbers of CD4+T, CD8+T and CD19+CD5+CD1d+B cells in mice form Blimp-1CKO group were higher than those of the control group (P<0.05), however, no significant differences with the percentages of these cell populations were observed between two groups.Higher numbers and percentages of CD19+CD5+B cells were detected in mice from Blimp-1CKO group (P<0.01).The Blimp-1CKO mice showed increased secretion of IFN-γ, TNF-α, IL-17 and IL-2, but decreased expression of CCR7 on CD8+T cells as com-pared with WT mice (P<0.05).No significant differences with the changes of S1P1 were found between the two groups.Conclusion Blimp-1 played an important role in the maintenance of number, phenotype and function of T cells.Furthermore, not only T cells but also B cell subsets in mice were affected by the dele-tion of Blimp-1 in T cells.

Objective To investigate the relationship between total homocysteine (tHcy) and carotid intima-media thickness (CIMT) in brain infarction patients. Methods Sixty patients with fasting plasma tHcy levels ≤10μmol/L (non-Hhcy group), 60 patients with fasting plasma tHcy levels>10μmol/L and≤15μmol/L (H1 group), and 60 patients with fast-ing plasma tHcy levels>15μmol/L (H2 group) were chosen in hospitalized patients with acute cerebral infarction. Values of CIMT were detected in three groups of patients. The clinical biochemical indicators including triglyceride (TG), total choles-terol (TC), high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C), fasting blood sugar (FBS), folic acid (FA), Vitamin B12 (VitB12) and glycated hemoglobin (HbA1c) were also detected. Results There was signifi-cant difference in CIMT between three groups (P<0.01). The value of CIMT increased in H2 group [0.98(0.90, 1.05)mm] com-pared with that of non-Hhcy group [0.85(0.80, 0.95)mm]. The value of CIMT increased in H2 group compared with H1 group [0.98(0.90, 1.05)mm vs 0.85(0.85, 0.95)mm], P<0.05). There were significant differences in tHcy, FA and VitB12 between three groups. Based on the log-transformed values of CIMT as the dependent variable, multiple stepwise linear regression showed significant associations of the following variables with increased CIMT: increasing age, the history of smoking, the history of diabetes, higher LDL-C and tHcy levels. Conclusion Brain infarction in patients with higher tHcy level often have lower levels of FA and VitB12, and increased CIMT. When the level of tHcy >15 μmol/L, there is more significantly higher level of CIMT. The increased CIMT level was associated with some cerebrovascular risk factors in patients with brain infarction.