Objective:To research PHA-CIK cells' phenotype and cytotoxicity. Methods: Using phytohemagglutinin(PHA)to stimulate peripheral blood mononuclear cells(PBMNCs) for 24 h,then cultured like incubating cytokine induced killers by traditional method. On 15th day examined its immunophenotype by flow cytometry and using MTT method to evaluate cytotoxicity. Results: The ratio of CD3+,CD8+, CD3+ CD8+ and CD3+ CD56+ cells were high.PHA-CK cells cytotoxicity was strong in some degree.Conclusion:PHA-CIK cells had strong cytotoxicity.The result provides an experimental basis for biotherapy.

AIM: To study the characteristic of liver X receptor alpha (LXRα), its target gene expression and cholesterol efflux in human macrophages treated with atorvastatin. METHODS: Human monocyte-derived macrophages were collected and cultured. Macrophages were treated with or without atorvastatin. Apolipoprotein A-I mediated human monocyte-derived macrophage cholesterol efflux was detected by liquid scintillation counting method. Reverse transcription-polymerase chain reaction (RT-PCR) was used to evaluate the mRNA expression of LXRα and some of its target genes ABCA1, SREBP2, CETP, PLTP, apoE, MMP-9 and MIP-1α. The protein expression of LXRα, ABCA1, MMP-9 and MIP-1α was determined by Western blotting. RESULTS: Pre-incubation of human monocyte-derived macrophages with atorvastatin dose dependently (1-2 μmol/L) stimulated cholesterol efflux mediated by apolipoprotein A-I. Atorvastatin also increased the mRNA expression of LXRα, ABCA1, SREBP2, CETP, PLTP, and protein expression of LXRα, ABCA1, but decreased the expression of MMP-9 and MIP-1α at both mRNA and protein levels. CONCLUSION: Atorvastatin enhances the cholesterol efflux, upregulates LXR and some genes associated with cholesterol metabolism and inhibits inflammatory responses in macrophages, indicating that statins may affect the formation of foam cells by activating LXR signaling pathway.

Objective To study the effects of Wortmannin, inhibitor of PI3K and SB202190, inhibitor of P38 MAPK and PD98059, inhibitor of ERK1/2 on the migration of epidermal growth factor (EGF)-induced vascular smooth muscle cells (VSMCs). Methods There were fives groups in this experiment, including control group, EGF group, PD98059 (PD) group, SB202190(SB) group and Wortnannin (WT) group. The migration rate of the VSMCs was measured by wound healing assay. Results At the 24th hours after wounding, there was obvious migration in EGF group compared to control group. The migration of VSMCs was significantly inhibited in PD group, SB group and WT group compared to the EGF group, but there were no significant difference among three inhibitor groups. At the 30th hours after woun-ding, there was still obvious migration in EGF group compared to control group. The migration of VSMCs was significantly inhibited in the three iuhibitors group compared to the EGF group, and there were significant difference among three inhibitor groups. Furthermore, inhibiting effect on VSMCs in SB group was more obvious compared to PD group and WT group. Conclusion These results suggested that the migra-tion of EGF-induced VSMCs may play a role through PI3K, P38 MAPK and ERKI/2 signal pathways, and the effect of P38 MAPK signal pathway is very important.

BACKGROUND:The CD4+CD25+FOXP3+Treg cells have immunosuppression effect, and it is speculated that these cells may restrain the occurrence of acute graft-versus-host disease. OBJECTIVE:To observe the variety of the CD4+CD25+FOXP3+Treg cells in the peripheral blood from donors before and after granulocyte colony stimulating factor mobilization, and study the relationship between CD4+CD25+FOXP3+Treg cells and acute graft-versus-host disease. METHODS:Ninety patients with malignant blood diseases who undertook al ogeneic hematopoietic stem celltransplantation and their donors were observed. Granulocyte colony stimulating factor 5μg/kg was injected subcutaneously into the donor per 12 hours for 5 days, and the stem cells were col ected before and after mobilization. The ratio of CD4+CD25+FOXP3+Treg cells in the peripheral blood was detected before and after mobilization with flow cytometry, and the ratio of these cells in the stem cellsuspension was measured by the same method. The patients were divided into two groups according to the CD4+CD25+FOXP3+Treg cells ratio:high dosage group (≥5%) and low dosage group (<5%). The incidence of acute graft-versus-host disease was observed in the two groups after transplantation. RESULTS AND CONCLUSION:The ratio of the CD4+CD25+FOXP3+Treg cells in the donor before and after mobilization were 11.3%and 1.5%,respectively, and there was a significant difference (P<0.05). The ratio of the CD4+CD25+FOXP3+Treg cells was 3.4%in the patients with acute graft-versus-host disease, and 15.7%in the patients with no acute graft-versus-host disease, showing a significant difference (P<0.05). After hematopoietic reconstitution, the incidence of acute graft-versus-host disease was 18.4%in the high dosage group and 48.1%in the low dosage group, and there was a significant difference between the two groups (P<0.05). Therefore, the granulocyte colony stimulating factor can lower the ratio of CD4+CD25+FOXP3+Treg cells in the human peripheral blood. The increase in CD4+CD25+FOXP3+Treg cells can restrain the occurrence of acute graft-versus-host disease.

ObjectiveTostudycellularimmunefunctioninpatientswithsyphilis.MethodsUsingflowcytometry,thecharacteristicofperipheralbloodlymphocyteimmophenotypes(PBLI)of23casesofsyphilis,andtherelationshipbetweenthePBLIandtherapeuticeffectwereanalyzed.Meanwhile,serumlevelofsIL-2Rofthepa-tientswasdetectedbyELISA.ResultsTheresultsshowedthatthepercentagesofpanT,TH,NKcellsandtheratioofTH/TSwerelower(P

Background:Gastric xanthelasma is a benign and uncommon lesion with unknown etiology. It has been noted thatatrophic gastritis is frequently seen in patients with gastric xanthelasma. Aims:To investigate the relationship betweengastric xanthelasma and atrophic gastritis and its clinical significance. Methods:A total of 10645 consecutive patients whohad undergone gastroscopy from Apr. 2015 to Mar. 2016 at the Shanxi Provincial People's Hospital were enrolled andanalyzed retrospectively with respect to their demographic,clinical,endoscopic and histological features. Results:Theprevalence of gastric xanthelasma in patients recruited in this study was 2. 9％ . The mean age and prevalence of atrophicgastritis (47. 9％ vs. 16. 6％ )were significantly higher and the gastric atrophy was more severe in patients with gastricxanthelasma than those without (P all < 0. 001). Multivariate Logistic regression analysis revealed that age ≥50 years(adjusted OR =1. 349,95％ CI:1. 042 ～ 1. 747,P = 0. 023)and atrophic gastritis (adjusted OR = 3. 892,95％ CI:3. 076 ～4. 924,P <0. 001)were independently related to the presence of gastric xanthelasma. Likewise,the prevalence ofgastric xanthelasma in patients with atrophic gastritis was significantly higher than those without (7. 9％ vs. 1. 8％ ,P <0. 001). Age- and sex-matched control analysis proved the correlation between gastric xanthelasma and atrophic gastritis(P <0. 001). In addition,multiple xanthelasma was more prevalent in gastric xanthelasma patients with atrophic gastritisthan those without (32. 0％ vs. 13. 8％ ,P <0. 001). Conclusions:Gastric xanthelasma is correlated with age and presenceand severity of atrophic gastritis. Atrophic gastritis might be one of the etiological factors of gastric xanthelasma,and gastricxanthelasma might be an atypical mucosal lesion caused by gastric atrophy.

This study examined the effect of ischemia-reperfusion injury on the expression of Pim-3 gene in myocardial tissues and their underlying mechanism. Rat models of myocardial ischemia-reperfusion injury were established by ligating the left anterior descending coronary artery of the rats. A total of 30 SD male adult rats were randomly divided into 5 groups: group A (sham operation, n=6); group B (in which the rats were subjected to 15 min of ischemia by ligation of the left anterior descending coronary artery, n=6); group C (in which the rats received 30 min of ischemia, n=6), group D and group E (in which the left anterior descending coronary artery of the rats were ligated for 30 min and then reperfused for 30 min or 120 min, n=6 in each). The left ventricular tissues were removed immediately after the ischemia-reperfusion injury. Neonatal cardiomyocytes were cultured and treated with different concentrations of H(2)O(2) (0, 5, 10, 20 μmol/L) or tumor necrosis factor-α (TNF-α, 0, 1, 5, 10 ng/mL). The mRNA and protein expression of Pim-3 gene was determined by using RT-PCR, western blotting and immunohistochemistry. Additionally, neonatal cardiomyocytes were transfected with Pim-3 siRNA, and induced to develop apoptosis by using H(2)O(2). The results showed that normal myocardial tissues expressed a quantity of Pim-3 gene mRNA and protein. Ischemia-reperfusion injury could up-regulate the mRNA and protein expression of Pim-3 gene in myocardial tissues. Furthermore, H(2)O(2) but not TNF-α up-regulated the Pim-3 gene expression in cultured cardiomyocytes. And Pim-3 silencing failed to strengthen the H(2)O(2)-inducing apoptosis in cardiomyocytes. It was concluded that ischemia-reperfusion injury up-regulated the Pim-3 gene expression through oxidative stress signaling pathway in myocardial tissues.

BACKGROUND: Transplantation of bone marrow mesenchymal stem cell and vascular endothelial growth factor(VEGF) can promote vascular regeneration and improve heart function. However, whether the combined application is superior to single application or not is still unclear.OBJECTIVE: To observe the effect of allogenetic bone manow stem cells transplantation combined with VEGF transfection on vascular regeneration and heart function of rats with acute myocardial infarction.DESIGN: Simple sample observation was used in culturing bone marrow mesenchymal stem cell of rats; Randomized controlled animal experiment was used in cell transplantation and gene transfection.SETTING: Department of Cardiology, Union Hospital of Huazhong University of Science and Technology; Cardiovascular Institute of Tongji Medical College MATERIALS: Totally 94 healthy male Wistar rats and expression vector PAdTrack/VEGF165 were used in this experiment.METHODS: This experiment was carried out at Cardiovascular Institute of Tongji Medical College between June 2004 and June 2005. ①Bone marrow mesenchymal stem cells of rats were isolated, purified and cultured in vitro, then labeled with bromodeoxyuridine(BrdU). ② Preparation , extraction, purification and identification of plasmid PAdTrack/VEGF165. ③Two weeks after coronary artery was ligated to create acute myocardial infarction model, rats were randomly divided into 4 groups (n=12 in each group): stem cell + plasmid group(50 μL BrdU-labeled bone marrow mesenchymal stem cell solution and 100 μL plasmid PAdTrack/VEGF165 were injected into the rats through multiple sites), stem cell group (50 μL bone marrow mesenchymal stem cell solution was injected through multiple sites), plasmid group (100 μL plasmid PAdTrack/VEGF165 was injected through multiple sites) , control group(100 μL serum-free DMEM was injected through multiple sites). ④ Immunohistochemistry andechocardiography were performed 4 weeks later.MAIN OUTCOME MEASURES: ①Immunohistochemical and haematoxylin-eosin stainings were conducted in the infarcted and ischemic areas of rats in each group; ② Blood vessel counts; ③Echocardiography.RESULTS: Totally 48 rats entered the stage of result analysis. ① BrdUlabeled transplanted cells could be seen at the infarcted and ischemic myocardium in the stem cell+plasmid group and stem cell group. Some transplanted cells at ischemic myocardium differentiated into vascular endothelial cells and formed newborn blood capillary. ②Density of Ⅷ factor positively-stained newborn blood capillary took stem cell +plasmid group ＞ plasmid group ＞ stem cell group ＞ control group in order (all P＜ 0.01).③Wall thickness and wall motion range improved after cell transplantation and gene transfection therapy. The increased range of ejection fraction took stem cell +plasmid group ＞ stem cell group ＞ plasmid group ＞ control group in order (all P ＜ 0.01) .CONCLUSION: Allogenic bone marrow mesenchymal stem cell transplantation and VEGF gene transfection could further boost vascular regeneration of infarcted ischemic area and improve wall thickness and heart function of rats.

In order to improve the efficacy of modified inferior method or middle method of radiofrequency catheter ablation (RFCA) in the treatment of atrioventricular node reentrant tachycardia (AVNRT), the clinical data of 325 cases of AVNRT from March 1992 to Feb. 2000 being subjected to the treatment of RFCA were retrospectively analyzed. The results showed that the successful rate was increased and recurrence was decreased year by year. In the recent 4 years the effective rate was up to 100 %. The complication of three grade of AVB occurred in 3 % and recurrent rate in 9.1 % before March 1996, but both of them were zero in the last 3 years. The time of RFCA procedure and X-ray exposure was significantly reduced. It was concluded that ablating more than 3 targets by modified inferior method or middle method with energy titrating and strict endpoint was the crux of obtaining satisfactory therapeutic effects and preventing recurrence.

In order to improve the efficacy of modified inferior method or middle method of radiofrequency catheter ablation (RFCA) in the treatment of atrioventricular node reentrant tachycardia (AVNRT), the clinical data of 325 cases of AVNRT from March 1992 to Feb. 2000 being subjected to the treatment of RFCA were retrospectively analyzed. The results showed that the successful rate was increased and recurrence was decreased year by year. In the recent 4 years the effective rate was up to 100 %. The complication of three grade of AVB occurred in 3 % and recurrent rate in 9.1 % before March 1996, but both of them were zero in the last 3 years. The time of RFCA procedure and X-ray exposure was significantly reduced. It was concluded that ablating more than 3 targets by modified inferior method or middle method with energy titrating and strict endpoint was the crux of obtaining satisfactory therapeutic effects and preventing recurrence.