Objective To study the changes of biological characteristics of CD34~+/CD133~+/VEGFR-3~+ lymphatic endothelial progenitor cells(EPCs) isolated from umbilical cord blood after induction with VEGF-C and investigate the mechanisms of EPCs differentiation toward lymphatic endothelial cells. Methods Mononuclear cells were isolated from umbilical cord blood using density centrifugation with Percoll solution.CD34~+/CD133~+/VEGFR-3~+ cells were sorted with FACS.Differentiation of the cells was induced with VEGF-C.The ultrastructural changes of the cells were viewed under scanning and transmission electron microscopes.Changes in expression of markers of the cells were viewed under confocal laser scanning microscope. Results Lymphatic EPCs isolated from umbilical cord blood expressed CD34,CD133 and VEGFR-3.At 7 day after induction with VEGF-C,CD34~+/CD133~+/VEGFR-3~+ cells were shuttle-like,there were a lamellipodium,numerous filopodia and many short microvilli.Caveolae were observed.Mitochondrion and rough endoplasmic reticulum were rich in cytoplasm.At 14 day after induction,the cells demonstrated appearance of the endothelial cell and expressed lymphatic endothelial specific makers LYVE-1 and 5'-nucleotidase,the expression of CD133 disappeared.Weibel-Palade body was observed in the cytoplasm of the cell.Conclusion There are CD34~+/CD133~+/VEGFR-3~+ lymphatic EPCs in human umbilical cord blood.The cells may differentiate into lymphatic endothelial cells through VEGF-C/VEGFR-3 signaling pathway after induction with VEGF-C.

Objective To explore college students’ sleep and insomnia syndrome type of traditional Chinese medicine.Methods A questionnaire survey was conducted among 1 325 college students of independent colleges using the Pittsburgh sleep quality index and the standardization of TCM syndrome introduced by guiding principles of clinical research on treatment of insomnia with new drugs of traditional Chinese medicine (2002).The results underwent analysis.Results Of 1 325 college students in independent college,209 people had poor sleep quality,in which liver fire retarding type accounted for 129 people,hyperactivity of fire due to yin deficiency type accounted for 67 people,followed by the heart and spleen deficiency type two(3 people),phlegm heat disturbance type(4 people),Qi deficiency type (2 people),other type (4 people).Conclusions College students in sub-health state with insomnia showed two common types:liver depression transforming into fire,hyperactivity of fire due to yin deficiency type.We should give effective intervention based on dialectical theory of the Chinese medicine.

Objective To investigate the incidence of target organ damage in prehypertensive. Methods Two hundred and seventy-eight prehypertensive cases (prehypertensive group) and 136 normotensive cases(normotensive group) were involved in this study. The blood pressure,body height,weight and waist circumference were measured. Serum creatinine (SCr), urea nitrogen (BUN),uric acid(UA), fasting blood glucose (FBG), total cholesterol (TC), triglyceride(TG),low density lipoprotein cholesterol (LDL-C), high density lipoprotein cholesterol (HDL-C) and morning urine albumin concentration were detected by automatic biochemical analyzer. Left ventricular end-diastolic dimension (LVEDd), left ventricular posterior wall thickness (LVPWT), interventricular septal thickness (IVST), intima-media thickness (IMT) and plaque index (PI) were examined by ultrasonography. The incidence rates of plaque, left ventricular mass index (LVMI), relative wall thickness (RWT), glomerular filtration rate (GFR) and body mass index (BMI) were calculated. Results The proportion of ≥ 1, ≥2 and ≥3 kinds of risk factors was 71.9% (200/278),40.6%(113/278) and 13.7%(38/278) in prehypertensive group and 30.1%(41/136), 13.2% (18/136) and 5.9%(8/136) in normotensive group, and there was significant difference between the two groups (P＜0.05). Compared with normotensive group, the age,systolic pressure, diastolic pressure, FBG, waist, BMI, morning urine albumin, TG,TC,UA,LDL-C,IVST,LVPWT,RWT,LVMI,IMT,PI were higher, meanwhile the incidence rate of concentric reconstruction of left ventricular, concentric hypertrophy, eccentric hypertrophy and carotid plaque was higher, and the incidence rate of left ventricular normal configuration and GFR was lower (P＜0.01 or ＜0.05). Conclusion People with prehypertensive may accompany with many kinds of cardiovascular risk factors, and the incidence rate of target organ damage is significantly increased.

Objective To investigate effects of endothelial adhesion molecule platelet endothelial cell odhesion molecule-1(PECAM-1),intercellular adhesion molecule-3(ICAM-3) and CD44 on lymphangiogenesis. Methods The lymphatic endothelial cells of the canine thoracic ducts were isolated and incubated.PECAM-1,ICAM-3 and CD44 on the lymphatic endothelial cells were labeled and visualized under fluorescence microscope and confocal laser scanning microscope.The endothelial cells stimulated by TNF-? or LPS were treated with the blocking antibodies against PECAM-1, ICAM-3 and CD44 respectively.The proliferation and migration of cells were determined with cell counting. The three-dimensional collagen gel model of lymphangiogenesis was prepared, and then the tube formation was viewed. The total length and area of the tubes formed from lymphatic endothelial cells were measured and their characteristics were examined with a transmission electron microscope. Results PECAM-1, ICAM-3 and CD44 were expressed on the lymphatic endothelial cells. In the control, TNF-? and LPS groups, the percentage of the migrated cells decreased and the total length and area of the tubes reduced after PECAM-1, ICAM-3 and CD44 of the cells were blocked respectively. The proliferated cells reduced after PECAM-1 or CD44 of the cells were blocked. However, there were no significant changes in the numbers of proliferated cells when ICAM-3 of the cells was blocked. In the semi-ultrathin and ultrathin sections, the tube structures formed by lymphatic endothelial cells were similar to lymphatic capillary.Conclusion PECAM-1, ICAM-3 and CD44 are expressed on the cultured lymphatic endothelial cells. These adhesion molecules are involved in lymphangiogenesis that includes proliferation, migration and tube formation of endothelial cells.

The training of airway management plays a vital role in standardized residency training plan during the rotation in the department of anesthesiology.Problem based-learning and airway simulation practice should be applied step by step based on the different requirements of standardized residency training plan.The training should be started from mask ventilation to endotracheal intubation,meanwhile clinical thinking ability and teamwork ability should be cultivated for the residents so that they can master the technique of airway management skillfully whenever and wherever.

Objective Compared with the effect of thromboembolectomy assisted with intraoperative digital subtraction angiography (DSA) and traditional method,evaluate the value of intraoperative DSA on surgical procedures for acute lower limb arterial embolism. Methods Sixeight cases of traditional thromboembolectomy to take on the operation (group A) and 72 cases under the DSA operation (group B)were analyzed during January 2005 to December 2009, Comparing two sets of operation (time, the amount of bleeding and the ankle brachial index changes before and after the surgery , amputations rate). Results The operation time of group B [(76 ±17) min] was less than that of group A [(95 ± 22) min, t =5. 736, P ＜ 0. 01] ; the amount of bleeding of group B [(83 ± 35 ) ml] was significantly less than group A [(102 ± 58 ) ml, t = 2. 362, P ＜ 0. 05]; The difference of ABI in group B after the operation than before (0. 32 ±0. 08) than that in group A(0. 25 ±0. 12) had remarkably improved ( t =2. 33, P ＜0. 05) ;there were 8 patient amputations in group A and 2 in group B ( u = 2. 06 , P＜ 0. 05 ); there were 2 patients died in group A and one in group B ( P ＞ 0.05 ). Conclusion Thromboembolectomy assisted with DSA can be accurate assessment of the arteries embolism position, elevate the success rate in the operation ,reduce the rate of amputation saws.

Objective To investigate the effects of VEGF-C,SDF-1,MCP-1 and G-CSF on migration and mobilization of CD34~+/CD133~+/VEGFR-3~+ lymphatic endothelial progenitor cells(LEPCs).Methods Mononuclear cells were isolated from canine peripheral blood by non-continuous density centrifugation with Percoll solution.VEGFR-3~+ cells were sorted with flow cytometxy.Expression of specific marks CD34 and CD133 on VEGFR-3~+ cells were observed under a confocal laser scanning microscope.The effects of VEGF-C,SDF-1,MCP-1 and G-CSF on chemotatic migration of LEPCs were examined by transmigration assay.The morphological characteristics of the transmigrated cells were observed under scanning electron microscope.VEGF-C,SDF-1,MCP-1 and G-CSF were administrated to rats by subcutaneous injection.The number of LEPCs in peripheral blood was analyzed with flow cytometry and white blood cells were counted with blood cell analyzer.Results The percentage of the migrated cells through the membrane was greater in the VEGF-C,SDF-1,MCP-1 and G-CSF groups than that in the control group.After the cells were treated with blocking antibodies of VEGFR-3 and CXCR-4,the percentage of the migrated cells decreased obviously.After the injections of VEGF-C,SDF-1,MCP-1 and G-CSF,LEPCs in peripheral blood of rats increased obviously,while significant increase of white blood cells was observed.Conclusion VEGF-C,SDF-1,MCP-1 and G-CSF play a role in the chemotatic migration and mobilization of LEPCs.The signaling pathways of VEGF-C/VEGFR-3 and SDF-1/CXCR-4 have important regulating effects on the chemotatic migration and mobilization of LEPCs.

AIM:To investigate the changes of autophagy of macrophages after phagocytizing dust particles and explore the mechanisms of dust particle-induced autophagy of the cells.METHODS:The bronchopulmonary lymph nodes were collected from patients with lung operation.The paraffin sections were prepared and then stained with Wilder's method.The tissue structures were viewed.Peritoneal macrophages were harvested from rats and then treated with carbon particles.Influences of carbon particles in autophagic activities of macrophages were examined.The ultrathin sections of the lymph nodes and the cells phagocytized carbon particles were prepared.The structures and distribution of phagosomes,autophagosomes and lysosomes were viewed.The apoptotic cells in the dust cells of the lymph nodes and the cells having phagocytized carbon particles were examined using transmission electron microscope and TUNEL staining.RESULTS:In adult lymph nodes,dust particles were deposited significantly in macrophages,collagen fibres and density of microvessels increased.There were autophagosome precursors,autophagosomes,autophagolysosomes as well as phagosomes in the dust cells and the cells phagocytized carbon particles.In autophagosomes,mitochondrion,dust particle or carbon particle were usually observed.There were positive cells by TUNEL staining in the dust cells and the cells phagocytized carbon particles.Nuclear condensation or apoptotic body in the apoptotic cells were observed under transmission electron microscope.CONCLUSION:Deposition of dust particles induces enhancement of autophagic activities and apoptosis of macrophages.Autophagy plays an important role in cleaning dust particles and the injured mitochondria.

Objective To study the microstructural characteristics of white matter fiber tracts of visual pathways using diffusion tensor imaging (DTI)in the patients with primary glaucoma.Methods Twenty patients with primary bilateral glaucoma and 30 nor-mal adults were recruited and scanned with visual pathways magnetic resonance imaging (MRI)and DTI,obtaining T1 FLAIR ima-ges and directionally encoded color (DEC)images.On DEC images,ROIs were put in the anterior,middle and posterior part of bi-lateral optic nerve,optic tract and optic radiation,to calculate the average FA and MD value of each region.Results T1 FLAIR and DEC images could clearly show the optic nerve,optic tract and optic radiation.In patients with glaucoma,the FA values of optic nerve,optic tract and optic radiation were:0.372±0.040,0.340±0.036,0.31 5±0.026,while the MD values were:1.760±0.1 1, 1.831±0.09,1.927±0.10.The FA values of optic nerve,optic tract and optic radiation of normal adults were:0.538 ±0.073, 0.460±0.082,0.455±0.083,whilst the MD values were:0.995 ±0.27,1.312 ±0.40,1.347 ±0.37.Compared with the normal controls,FA values of all parts are deceased whereas the MD values of all parts are increased in patients.Conclusion Glaucoma is a central nervous system disease involving the whole visual pathways.Use DTI can evaluate the microstructural characteristics of white matter fiber tracts of visual pathways in the patients with glaucoma.

Objective: To study the pharmacology of Zhiloushu Pills (Radix et Rhizoma Rhei, Stiff silk worm and Flos Sophorae). WTHZ]MethodsMethods: Zhiloushu Pills, clysterred into animals. Results: Zhiloushu Pills can decrease the inflammatory caused by CMC. Conclusion: Zhiloushu Pills are helpful to heal the rabbits local infection caused by staphylococcus aureus. They can also shorten the blood coagulation time and have the function of invigorating the circulation of blood and removing the stasis of blood against stasis animals.