Objective To investigate the eftect of VEGF gene transtection on endothelial progenitor cell derived from murine bone marrow.Methods Wistar rat's bone marrow was obtained, mononuclear cell isolated,and endothelial progenitor cells(EPS)were cultured in EGM-2MV.EPCs were identified by immunocytochemistry and electron microscope.EPCs were transfected by liposome mediated pcDNA3.0-hVEGF165.VEGF protein level was determined in the cultural medium supernatant after VEGF transfection by ELISA.Cultural medium supernatant was used to co-culture with ECV304,VEGF protein activity was evaluated by MTT.EPCs expression of vWF,VEGF,FLK-1 was detected by immunocytochemistry.Results EPCs were effectively enriched by EGM-2MV,and the EPCs obtained express the typical cell surface markers such as CD34,CD133,FLK-1.The concentration of VEGF protein in supernatant reaches 1280 pg/ml in the 7th day after pcDNA3.0-hVEGF transfection.No influence of EPCs proliferation could be found after transfeetion.The cell surface marker expression of VEGF,FLK-1, vWF became higher with time,and the ratios of positive cell were 88.52%,82.65% and 95.97% respectively.Conclusions pcDNA3.0-hVEGF165 transfeet EPCS mediated by liposome could excrete a high concentration of functional VEGF protein.It is helpful for EPC to maintain the characters of endothelial cell after VEGF gene transfection and differentiate to mature endothelial cell.

BACKGROUNDEndothelial progenitor cells (EPCs) have been used in both experimental studies and clinical treatments of limb ischemia, as well as in the construction of engineered vascular tissue. The objective of this study was to investigate the effects of transplanted bone marrow-derived EPCs on the vein microenvironment in a rat model of chronic vein thrombosis.

METHODSMononuclear cells were isolated from the bone marrow of immature rats by density gradient centrifugation, cultured, and then transplanted into experimentally induced thrombi into inferior vena cava through the femoral vein. Vascular endothelial growth factor (VEGF), angiopoietin-1 (ANG-1) and monocyte chemotactic protein-1 (MCP-1) mRNA and protein expression levels were measured by real-time quantitative polymerase chain reaction and Western blotting of thrombi and adjacent caval walls 28 days post-transplantation.

RESULTSLevels of VEGF, ANG-1, and MCP-1 mRNA in EPC-transplanted thrombi were 100%, 230.7%, and 212.5% of levels detected in the sham-operated group (P < 0.01), and 99.9%, 215.4%, and 177.8% of levels detected in the experimental control group (P < 0.01). VEGF, ANG-1 and MCP-1 protein levels exhibited a similar trend.

CONCLUSIONSTransplanted bone marrow-derived EPCs appear to alter the vein microenvironment in experimentally induced chronic vein thrombosis by upregulating cytokines associated with thrombic organization and recanalization.

Angiopoietin-1 ; analysis ; genetics ; Animals ; Bone Marrow Cells ; cytology ; Chemokine CCL2 ; analysis ; genetics ; Chronic Disease ; Disease Models, Animal ; Endothelial Cells ; cytology ; Fluorescent Antibody Technique ; Immunohistochemistry ; RNA, Messenger ; analysis ; Rats ; Stem Cell Transplantation ; Vascular Endothelial Growth Factor A ; analysis ; genetics ; Venous Thrombosis ; therapy

OBJECTIVETo assess the changes of somatosensory evoked potentials (SEPs) during spinal cord ischemia and reperfusion injury and the value of SEP monitoring in evaluating neurological functions in this setting.

METHODSSpinal cord ischemia-reperfusion injury was induced in 28 rabbits by clamping the infrarenal aorta for 45 min, and the SEPs were monitored before and at 5, 10, and 15 min after ischemia, and at 15, 30, and 60 min and 2, 24 and 48 h after reperfusion. The neurological function score (NFS) of the rabbits was evaluated at 6, 12, 24 and 48 h after reperfusion, and the pathological changes of the spinal cord were observed 48 h after reperfusion.

RESULTSSEPs P1-wave latency significantly increased 5 min after ischemia (P<0.01) and the wave amplitude decreased 8 min after ischemia (P<0.01). SEPs disappeared 10 min after ischemia and recovered 15 min after reperfusion, but the P1-wave latency still remained longer and P1-wave amplitude lower than the measurements before ischemia (P<0.01). P1-wave amplitude became normal 15 min after the reperfusion (P>0.05), and the P1-wave latency gradually recovered 30 min after reperfusion, but still significantly longer than the preischemic value (P<0.01). P1-wave amplitude decreased again at 24 and 48 h after reperfusion (P<0.01). The NFS gradually increased at 24 and 48 h after the reperfusion (P<0.01). The changes in P1-wave amplitude at 24 and 48 h after reperfusion showed an obvious correlation to NFS (r=-0.881 and -0.925, respectively, P<0.01). Hemorrhage, swelling, and degeneration and neutrophil infiltration occurred in the spinal cord tissue 48 h after the reperfusion.

CONCLUSIONThe changes of SEP P1-wave amplitude can better reflect the spinal cord function than the wave latency during spinal cord ischemia-reperfusion injury, and SEP monitoring provides reliable evidence for prognostic evaluation of the neurological function.

Animals ; Evoked Potentials, Somatosensory ; physiology ; Female ; Ischemia ; physiopathology ; Male ; Monitoring, Physiologic ; methods ; Rabbits ; Reperfusion Injury ; physiopathology ; Spinal Cord ; blood supply

BACKGROUNDMammalian target of rapamycin (mTOR) is involved in a caspase independent form of programmed cell death called autophagy. The aim of this research was to investigate the effects of rapamycin and 3-methyladenine (3-MA) on autophagy, proliferation, apoptosis, and cell-cycle parameters of rat bone marrow-derived endothelial progenitor cells (EPCs).

METHODSMononuclear cells isolated from rat bone marrow were treated with rapamycin (0.01, 0.1, 1, or 10 µg/L) or 3-MA (1.25, 2.5, 5, or 10 mmol/L) for 24 hours. Expression of the autophagy marker protein LC3-II was analyzed by Western blotting. Apoptosis and cell-cycle progression were analyzed by flow cytometry. Cell proliferation was measured using the MTT assay.

RESULTSRapamycin treatment of EPCs induced apoptosis and autophagy and inhibited proliferation and cell-cycle progression in a dose-dependent manner. Treatment with 5 mmol/L 3-MA promoted cell proliferation; in contrast, treatment with 10 mmol/L 3-MA promoted apoptosis and induced S-phase arrest.

CONCLUSIONSRapamycin treatment of EPCs induced apoptosis and autophagy. Low concentrations of 3-MA had no significant effect on the proliferation and apoptosis of EPCs; The 5 mmol/L group promoted cell proliferation, but had no effect on the apoptosis; the 10 mmol/L group inhibited the proliferation and promoted apoptosis through the cell cycle.

Adenine ; analogs & derivatives ; pharmacology ; Animals ; Apoptosis ; drug effects ; Autophagy ; drug effects ; Cell Cycle ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Rats ; Sirolimus ; pharmacology

Objective To study the mlationship between dietary soy isoflavones and blood lipids among residents of 40-65 years old,in Guangzhou.Methods Dietary soy isoflavones and other nutrients intakes were assessed with quantitative food frequency questionnaire(FFQ).Total cholesterol(TC),triglycerides(TG),HDL cholesterol(HDL-C)and LDL cholesterol(LDL-C)in plasma were measured with colorimetry.Results Ranges of dietary soy isoflavones intake among 134 males and 261 females were fxom 0 mg/day to 61.96 mg/day and 0 mg/day to 82.52 mg/day,with means of 11.95 mg/day,14.90 mg/day,respectively.After adjusted for total energy intake and fat percent energy,difiefences of TC,LDL-C in total population and TC in women were statistically significant between groups(P value was 0.002,0.008,0.004,respectively) and dose-effect relationships(P value was <0.001.0.012.0.001,respectively)were observed between dietary soy isoflavones intake and the upper mentioned three indices.Compared with the low-intake group,tbese three indices lowered 7.06%,10.13%and 7.48%,respectively in high-intake group.Critical significance of LDL-C was observed both in women and men between groups.Further controlled for age,BMI and WHR,no obvious change of the results was observed.Conclusion Moderate intakes of soy isoflavone as part of a regular diet seemed to be associated with favorable blood lipid levels.

Objective To analyze the motivation of Chinese patients with chronic myelogenous leukemia (CML) who have stopped the tyrosine kinase inhibitor (TKI). Methods Forty-seven CML patients who have stopped TKI provided informed consent prior to their participation in the study. These patients were divided into relapse and non-relapse group at the endpoint of the observation. None of the patients received any CML-associated therapies after TKI cessation. The reasons of withdrawal were analyzed statistically. Results The reasons for cessation included patient's request due to cost(59.57 %, 28/47), patient's plan to getting pregnant(8.52 %,4/47),side-effect of TKI(23.40 %,11/47)and other reasons(8.52 %,4/47).At the endpoint of observation, 23 patients suffered molecular relapse. Among them, 15 cases (65.22 %) were due to cost; 1 case (4.35 %) was due to getting pregnant, 5 cases (21.74 %) were due to side-effect and 2 cases (8.69 %) were due to other reasons. There was more frequency relapse in the group of insufficient cost. Conclusion The motivation of Chinese CML patients who have stopped TKI might show impact on the outcome,and the motivation is mainly related with history of drug reduction and withdrawal.

OBJECTIVETo study the analgesia mechanism of needle-knife lysis in spinal cord and other parts of central nervous system by comparing the changes of Leu-Enkephalin (L-ENK) content in different parts of centrium of rats undergone needle-knife lysis and electro-acupuncture respectively.

METHODSSixty healthy SD rats were randomly devided into normal control group, model group, needle-knife lysis (NKL) group and electro-acupuncture (EA) group. 4% papain solution mixed with 0.3 mol/L cysteine solutin in the ratio of 1:1, paused for 0.5 h,injected the mixture, 20 microl each time,into the left knee joint cavities of rats in model, NKL, EA groups at the 1st, 4th, 7th day. After 4 weeks in NKL group and EA group were treated with needle-knife lysis and electro-acupuncture, respectively. Three weeks after treatment, samples of spinal cord of the swollen part of rat waists and rat brains were taken from and the content of L-ENK of medulla oblongata, midbrain, pituitary gland, and hippocampus were measured.

RESULTSL-ENK content of model group increased higher than that of normal control group in spinal cord, hippocampus and midbrain (P < 0.01); there were no significant difference between normal control group and modle group on L-ENK in medulla oblongata and thalamus (P > 0.05). After intervening of NKL or EA, L-ENK content of NKL group increased higher in hippocampus than that of model group and EC group (P < 0.05); but L-ENK content of NKL group in midbrain was lower than that of model group (P < 0.05).

CONCLUSIONNeedle-knife lysis has characteristic of regulation for the L-ENK content in different parts of central nervous system of rats with knee osteoarthritis, and analgesic effect of needle-knife was possibly related with regulation of center L-ENK.

Acupuncture Therapy ; methods ; Animals ; Central Nervous System ; chemistry ; Electroacupuncture ; Enkephalin, Leucine ; analysis ; Female ; Male ; Osteoarthritis, Knee ; metabolism ; therapy ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo study the distribution of prevalence on hypercholesterolemia and borderline high cholesterol of Chinese residents aged 20 years and above.

METHODSTo detect the total fasting plasma cholesterol in 48 299 subjects among 20 year-olds and above.

RESULTSWithin a certain age range, the prevalence of both hypercholesterolemia and borderline high cholesterol increased with age, but there was a decreasing trend after 60-years of age in some regions. The prevalence of hypercholesterolemia increased to 12.53% in over 70 year-olds group from 1.36% in urban 20-year-olds, and increased to 5.15% from 60-year-olds group comparing to 0.98% of the 20-year-old group in rural areas. When comparing to the 20-year-old age group, the prevalence of borderline high cholesterol of the 70 age group in urban and rural increased by 4.4 and 4.2 times, respectively. The prevalence of hypercholesterolemia in each age group in urban were higher than that of in rural areas. In Rural I, it was higher than that of in Rural II , Rural III and Rural IV. The prevalence rates of both hypercholesterolemia and borderline high cholesterol of the males in the less than 50 year-olds group were higher than that in females, which was quite different from the group older than 50 years of age. As to borderline high cholesterol, similar result was also noticed.

CONCLUSIONWithin a certain age range, there was a trend that the prevalence of both hypercholesterolemia and borderline high cholesterol increased with age and related to regions which were classified by economic status. A distinct difference between males and females also existed.

Adult ; Age Factors ; Aged ; Asian Continental Ancestry Group ; China ; epidemiology ; Cholesterol ; blood ; Female ; Humans ; Hypercholesterolemia ; epidemiology ; Male ; Middle Aged ; Prevalence ; Rural Population ; Sex Factors ; Urban Population ; Young Adult

OBJECTIVETo investigate the role of mesenchymal stem cells (BMSCs) in multiple myeloma (MM) bone marrow (BM) microenrivonment and their effect on myeloma cells survival and bortezomib induced apoptosis.

METHODSBMSCs were derived from BM of untreated myeloma patients (MM-BMSCs) and healthy donors (HD-BMSCs), respectively. The phenotype, proliferation time and cytokine secretion of MM-BMSCs were detected and compared with HD-BMSCs. Then BMSCs were co-cultured with myeloma cell line NCI-H929 and bortezomib in vitro. The NCI-H929 cells proliferation and bortezomib induced cell apoptosis were investigated.

RESULTSMM-BMSCs and HD-BMSCs were isolated successfully. The phenotype of MM-BMSCs was similar to that of HD-BMSCs. Expressions of CD73, CD105, CD44 and CD29 were positive, but those of CD31, CD34, CD45 and HLA-DR (< 1%) negative. The proliferation time of MM-BMSCs was longer than that of HD-BMSCs (82 h vs 62 h, P < 0.05). Moreover, over-expressions of IL-6 and VEGF in MM-BMSCs culture supernatant were detected as compared with that in HD-BMSCs [(188.8 ± 9.4) pg/ml vs (115.0 ± 15.1) pg/ml and (1497.2 ± 39.7) pg/ml vs (1329.0 ± 21.1) pg/ml, respectively]. MM- BMSCs supported survival of the myeloma cells NCI-H929 and protected them from bortezomib induced cell apoptosis.

CONCLUSIONSMM-BMSCs is benefit for myeloma cells proliferation and against cell apoptosis induced by bortezomib. Over-expression of IL-6 and VEGF maybe play a critical role in these effects.

Apoptosis ; drug effects ; Bone Marrow Cells ; cytology ; Bortezomib ; Humans ; Mesenchymal Stromal Cells ; metabolism ; Multiple Myeloma ; metabolism

BACKGROUNDCatheter-directed thrombolysis (CDT) for deep venous thrombosis (DVT) of the lower extremity has good effect, but whether iliac vein stent placement after thrombolytic therapy is still controversial. The goal of this study was to evaluate the efficacy of stent placement in the iliac vein following CDT in lower extremity DVT.

METHODSThis was a single-center, prospective, randomized controlled clinical trial. After receiving CDT, the major branch of the distal iliac vein was completely patent in 155 patients with lower extremity DVT, and 74 of these patients with iliac vein residual stenosis of >50% were randomly divided into a control group (n = 29) and a test group (n = 45). In the test group, stents were implanted in the iliac vein, whereas no stents were implanted in the control group. We evaluated the clinical indicators, including patency of the deep vein, C in CEAP classification, Venous Clinical Severity Score (VCSS), and Chronic Venous Insufficiency Questionnaire (CIVIQ) Score.

RESULTSAll patients had postoperative follow-up visits for a period of 6-24 months. Venography or color ultrasound was conducted in subjects. There was a significant difference between the patency rate at the last follow-up visit (87.5% vs. 29.6%) and the 1-year patency rate (86.0% vs. 54.8%) between the test and control groups. The change in the C in CEAP classification pre- and post-procedure was significantly different between the test and control groups (1.61 ± 0.21 vs. 0.69 ± 0.23). In addition, at the last follow-up visit, VCSS and CIVIQ Score were both significantly different between the test and control groups (7.57 ± 0.27 vs. 0.69 ± 0.23; 22.67 ± 3.01 vs. 39.34 ± 6.66, respectively).

CONCLUSIONThe stenting of iliac vein obstruction following CDT in lower extremity DVT may increase the patency of the deep vein, and thus provides better efficacy and quality of life.

Adolescent ; Adult ; Aged ; Catheterization, Peripheral ; methods ; Female ; Humans ; Iliac Vein ; Lower Extremity ; pathology ; Male ; Middle Aged ; Stents ; Thrombolytic Therapy ; methods ; Venous Thrombosis ; therapy ; Young Adult