Three compounds were isolated from solid culture of endophyte Myrothecium roridum IFB-E091 in Artemisia annua. Their structures were determined as (S)-(-)-N-[2-(3-hydroxy-2-oxo-2,3-dihydro-1H-indol-3-yl)-ethyl]-acetamide (1), N-(4-hydroxyphenethyl)acetamide (2) and asperfumoid (3), in which compound 1 was a new indole derivative. In cytotoxicity assay, the compound 1 had no obvious inhibition activity in human hepatoma cell line SMMC-7721 and human cervical carcinoma cell line HeLa.

Objective To evaluate the correlations of emphysema and airway wall thickness to chronic obstructive pulmonary disease(COPD) of airflow limitation by quantitative CT.Methods 40 COPD patients and other 40 normal controls underwent pulmonary function tests and following MSCT exams with inspiration.The square root of wall area of an airway with an internal area of 8 mm2 (Ai8) and the percentage of low attenuation volume(LAV%) of the whole lung and each lobes at the threshold of-950 HU were measured by a software of Thoracic VCAR.The Ai8 between the observation group and the control one was compared using SPSS2.2.The contributions of LAV% and Ai8 to predictions of FEV1/FVC and FEV1% were also evaluated.Results There was a significantly statistical difference in the Ai8 between the observation group and the control one.There were correlations between airflow limitation markers and all of LAV% as well as Ai8 (P<0.05 for all standardized coefficients).Only the Ai8 of right inferior lobar made a significant contribution to airflow limitation in the whole lung bronchus, and the LAV% of each lobes made a stronger contribution to airflow limitation than the Ai8 of right inferior lobe.Conclusion There is a significantly statistical difference in the Ai8 between the observation group and the control one.The LAV% may make a greater contribution to airflow limitation than Ai8 in COPD group.The influential factors of airflow limitation in order were LAV%, Ai8 of right inferior lobe and Ai8 of the other lobes.

Objective To investigate the role of glycogen synthase kinase -3beta(GSK3β)on invasion and metastasis of human osteosarcoma cells .Methods Expression and phosphorylation of GSK 3βwere examined in osteosarcoma cell lines and tumor tissue from osteosarcoma patients by Western blot .The effects of small mole-cule GSK3βinhibitors on cell metastasis were detected by cell invasion assay and cell migration assay .Results Osteosarcoma cell lines showed increased GSK 3βexpression and abnormal activity regulation .In tumor tissue of patients with osteosarcoma metastasis and non -metastasis,GSK3βwere detected expression and abnormal activi-ty,especially in tumor tissue of the patients with osteosarcoma metastasis .Inhibition of GSK3βactivity resulted in inhibiting cells invasion and migration in osteosarcoma cell line .Conclusion In this research,we demonstrated that GSK3βencouraged osteosarcoma cells metastasis .The result will open up a potential target for clinical treat-ment of osteosarcoma .

Objective To study the repair function of united endothelial progenitor cells (EPC)transplantation on injured liver endothelium by bone marrow transplantation (BMT) conditioning.Methods C57BL/6 mice were divided into four groups randomly: normal control group, without any treatment; irradiation alone group, administered a total body irradiation(TBI) pretreatment, without BMT; (3) BMT alone group: C57BL/6 mice were infused with bone marrow mononuclearcells (MNC) 5 × 106/only through caudal vein not more than 4 h after the same TBI pretreatment as the irradiation alone group; united transplantation group: receiving the same way as the BMT alone group, but C57BL/6 mice were infused with EPC 5 × 105/only at the same time. Two, 4, 7, 14, and 21 days after the TBI, the changes of the liver weight were observed regularly. The histopathological examination of liver was done at the 4th, 7th, 14th, and 21st day after the TBI. Results In irradiation alone group, BMT alone group and united transplantation group the liver weight began to increase significantly on the day 2 and peaked at 14th day after the TBI, and the peaks were respectively (1.65±0. 15) times (P＜0. 05), (1.61 ±0.06) times (P＜0.05), and (1.11 ±0.40)times (P＜0. 05) of those in normal control group. At the day 14, the liver weight in irradiation alone group, BMT alone group and united transplantation group began to decrease, and on the day 21 the liver weight in united transplantation group had been completely restored to normal level, however the liver weight in irradiation alone group and BMT alone group were still significantly heavier than that in normal control group (P＜0. 05). Liver histopathological examination revealed that there were obvious sinusoidal endothelial cells (SEC) injury, hepatocyte edema and severe inflammatory cell infiltration in irradiation alone group, and on the day 7 the hepatocyte edema and necrosis were significantly worse than before, and almost no alive SEC were found. On the day 14 the injury of SEC in BMT alone group was lighter than before, but on the day 21 the injury had not returned to normal. On the day 7 the injury of SEC, hepatocyte edema and necrosis were alleviated in united transplantation group as compared with irradiation alone group and BMT alone group, and on the day 14 the injury had returned to normal basically. Conclusion The transplantation conditioning could damage recipient liver endothelium and the injury would persist, and united EPC infusion could repair the injured SEC following BMT.

Objective To determine the conditioning regimen suitable for mice allogeneic hematopoietic stem cell transplantation (allo-HSCT).Methods Twelve BALB/c mice were randomly divided into 2 equal groups to undergo X-ray irradiation by linear accelerator at the dose of 7.0 Gy (pure X-ray group) or 60Co source irradiation at the dose of 7.0 Gy (pure γ-ray group).Thirty mice were randomly divided into 2 equal groups to undergo X-ray irradiation and then infusion of bone marrow from donor mice via caudal vein (X-ray + transplantation group) or γ-ray and then infusion of bone marrow via caudal vein (γ-ray + transplahtation group).3,5,7,10,15,20,and 30 d later peripheral blood samples were collected to calculate the number of white blood cells (WBCs) and detect the chimeric rates of lymphocytes by flow cytometry.5,10,and 20 d after irradiation 15 mice were killed with their lung,liver,small intestine,spleen,and femurs taken out to undergo pathological examination.Results The survival rates during the period 5-15 days of the γ-ray + transplantation group were all significantly higher than those of the X-ray + transplantation group.The pathological changes of organs of the X-ray +transplantation group were all more severe than those of the γ-ray + transplantation group.Since the fifth day after transplantation cells originating from the donor began to appear in the peripheral blood.The chimeric rate of the γ-ray + transplantation group 10 days after transplantation was (95.53± 2.57) %.The chimeric rates 5,10,and 20 days after transplantation of the γ-ray + transplantation group were all significantly higher than those of the X-ray + transplantation group (t = 15.263,3.256,P < 0.05).The WBC count of both irradiation groups decreased to the lowest level 5 d later and began to increase 10 days after transplantation and the WBC counts of the γ-ray + transplantation group 10 and 20 days aftertransplantation were both significantly higher than those of the X-ray + transplantation group (t = 3.624,6.695 ,P < 0.05).The chimeric rats of the peripheral lymphocytes 10 and 20 days after transplantation of the γ-ray + transplantation group were both significantly higher than those of the X-ray + transplantation group (t = 12.317,8.295,P < 0.05).The homogeneity rate of transplantation of the γ-ray +transplantation group was better than that of the X-ray + transplantation group.Conclusions As a conditioning regimen in allogeneic hematopoietic stem cell transplantation γ-ray irradiation causes milder injury and accelerated reconstitution of hematopoiesis and immunity,in comparison with X-ray irradiation.

Objective To study the relationship between graft-versus-host disease (GVHD) and endothelium injury following hematopoietic stem cells transplantation in mice. Methods C57BL/6 mice as donors and Balb/c mice as recipients were randomly divided into 4 groups: control group, bone marrow transplantation group, GVHD group, GVHD mitigation group. The clinical manifestations,circulating endothelial cells and tissue pathological changes were observed at different time points after transplantation. Results No manifestations of GVHD were found in each group at the day 5, while those were found in GVHD group at the day 9 and all died within 15 days. The counts of endothelial cells in peripheral blood showed no significant difference at the day 5 between GVHD group (7. 34 ±1.26 cells/μl) and bone marrow transplantation group (11.51 ± 7. 40 cells/μl) or GVHD mitigation group (7. 36 ± 0. 16 cells/μl), while among three groups there was statistically significant difference at the day 9 (GVHD group: 153. 64 ± 35. 35 cells/μl vs bone marrow transplantation group: 10. 49 ±5. 61 cells/μl and GVHD mitigation group: 47. 82 ± 4. 69 cells/μl). The scores of pathological aGVHD had no significant difference at the day 5 between GVHD group (4. 33± 1. 53) and bone marrow transplantation group (3. 33 ± 0. 58) or GVHD mitigation group (4. 00 ± 1.73), while among three groups there was statistically significant difference at the day 9 (GVHD group: 10. 0 vs bone marrow transplantation group: 3. 33 ± 1.15 or GVHD mitigation group: 4. 33 ± 0. 58) and at the day 14 (GVHD group: 10. 33 ± 2. 58 vs bone marrow transplantation group: 2. 33 ± 1.25 or GVHD mitigation group 3. 33 ± 1.15). Conclusion Occurrence of GVHD causes endothelial damage again and injured endothelium worsens the GVHD.

Objective To (e)xplore inhibition of endothelial progenitor cells (EPCs) against hepatic vein thrombosis after allogeneic bone marrow transplantation (BMT).Methods Balb/c mice were randomly divided into three groups: (1) BMT group [Balb/c mice were injected intravenously with 5 × 106 bone marrow cells after total body irradiation (TBI)]; (2) EPCs co-transfusion with bone marrow cells group: 5 × 105 EPCs were infused into recipient mice simultaneously; (3) Normal control group.Liver index was detected on the day 0,5,10,15 and 20 after transplantation.Hepatic vein thrombosis,hepatic cells and vascular endothelial damage were observed under the light microscopy after H&E staining.The injury of liver cells,liver veins,hepatic sinusoidal endothelial cells (SECs)and platelet adhesion conditions were observed under a transmission electron microscope (TEM).The proportion of activated platelets and TNF-α concentration in peripheral blood were detected by using flow cytometry.Results On the day 0,5,10,15 and 20 after transplantation,the proportion of activated platelets,liver index and TNF-α concentrations in BMT group and EPCs co-transfusion group showed an upward trend,peaked on the 15th day,and then decreased.However,they were still significantly higher than those in normal control group (P＜0.05).The above parameters in EPCs co-transfusion group at each time point were significantly lower than those in BMT group (P＜0.05).As compared with BMT group,platelet adhesion decreased,hepatic vein thromboses were reduced,hepatocyte swelling and necrosis were alleviated,and liver damage repaired rapidly in EPCs co-transfusion group.Conclusion EPCs co-transfusion with bone marrow cells could inhibit the hepatic veins thrombosis and ameliorate liver damage significantly.

Aim To investigate the effects of saponin of sea cucumber ( SSC ) on the blood pressure in obese mice. Methods C57BL/KsJ(db/db) mice were ran-domized into 3 groups ( 8 mice each ): model group, low-dose SSC group and high-dose SSC group. Normal C57BL/KsJ mice were used as control. The low and high SSC groups were fed on basal diets incorporated with 0. 02% and 0. 04% SSC. Different treatments were administered for 6 weeks and arterial pressure was measured in the third and sixth weeks. The abundance of renal ACE, ACE2 and REN mRNA was detected by real time PCR . Results Compared with control group, the blood pressure of model group mice was ob-viously raised ( P<0. 01 ) . Low-dose SSC group mice showed lower blood pressure than model group without statistically significant differences, and the blood pres-sure of high-dose SSC group mice was similar to that of control group and significantly lower than model group. ( P<0. 05 ) There were no remarkable differences a-bout ACE and REN mRNA among the groups, howev-er, ACE2 mRNA level was significantly increased in high-dose SSC group. Conclusion SSC plays a vital role in decreasing blood pressure, which probably re-lates to the regulating function of renin-angiotensin sys-tem( RAS) .

Objective To explore a proper dose of endothelial progenitor cells (EPCs)administration that can achieve optimal hematopoietic improving effectiveness in a murine allogeneic hernatopoietic stem cell transplantation (allo-HSCT) model.Methods Female Balb/c mice were lethally irradiated with 60Co source,and then were injected intravenously with 5 106 bone marrow cells from C57BL/6 mice (bone marrow transplantation group).In co-transfer experiments,5 × 104,1 ×105,5 × 105 or 1 × 106 donor EPCs (EPCs treated groups) were injected simultaneously with bone marrow cells.The recipients were monitored for survival,peripheral white blood cells,hematopoietic stem cells (HSCs) and bone marrow histology.Results Compared with bone marrow transplantation group,all EPCs treated groups had accelerated recovery of peripheral white blood cells (P＜0.05),platelets (P＜0.05) and HSCs (P＜0.05).When infused with less than 5 × 105 EPCs,these effective hernatopoietic improving phenomena showed a positive correlation with the administrated doses of EPCs.However,when infused with 1 × 106 EPCs,the mice showed lower survival rate (P＜0.05)and slower recovery of peripheral white blood cells (P＜0.05),platelets (P＜0.05) and HSCs (P＜0.05) than 5 × 105 EPCs treated grpup.Bone marrow histopathology analysis confirmed the above findings.Conclusion Co-transfer with donor EPCs can improve survival rate and hematopoietic reconstitution of recipient mice in allo-HSCT,and 5 × 105 EPCs should be a proper dose to achieve the best effectiveness.

Objective To assess the functional role of TH 17 cells in allogeneic hematopoietic stem cell transplantation (allyHSCT).Methods Bone marrow monocytes and splenic T cells were enriched from C57/BL6 donors.Recipient Balb/c mice were irradiated with 7.5 Gy total body irradiation (TBI) and injected with 5 105 splenic T cells and 5 106 bone marrow monocytes.Survival was monitored daily,clinical graft-versus-host disease (GVHD) was assayed three times a week,and detailed histopathologic analyses of lung were performed at day six after Allo-HSCT.Flow cytometry analysis was performed using CD3-FITC,CD4-PE,CD45-PerCP-CyS.5 monoclonal antibodies.Cells were stained for intracellular cytokines using mouse TH 1/TH2/TH 17 cytokine kit.Results All the experimental animals showed GVHD manifestations on the day 6 after transplantation.Animals from BMT and HF groups were scarified and histological analysis of lung was performed.Absence of TH 17 cells induced severe pathologic pulmonary lesions.The histopathology of the lung tissue was characterized by disorganization,epithelia cell damage,interstitial fibroplasias,and monocytes infiltration.The proportion of TH1 and TH 17 in BMT group was (5.53 ± 0.11 ) ％ and ( 1.04 ± 0.34)％ respectively,both significantly different from that in HF group.The levels of IL-17A and IFN-γin BMT group were (2.81 ±0.19) and (42.97 ± 0.23) pg/mL respectively.IL-17A could not be detected in HF group,yet the level of IFN-y was only (9.89 ± 0.51 ) pg/mL.IL-10 in both HF and BMT groups was not detectable.Conclusion Lung is on target of aGVHD.IL-17A may play a key role in the lung injury after transplantation.