BACKGROUND: Vascular endothelial growth factor receptor 2 (VEGFR2) is primarily involved in vascular endothelial growth factor (VEGF)-mediated signal transduction and plays a critical role in the pathological angiogenesis that occurs in a number of diseases, including leukemia. Besides, VEGF secreted by leukemia cells also induces its own expression which leads to an enhanced production of VEGFR2 which contributes to the survival and proliferation of leukemia cells.OBJECTrVE: To evaluate the inhibitive effect of Lenti6/shVEGFR2 on the VEGFR2 expression and leukemia growth in mouse.DESIGN: A randomized, parallelized, controlled and open trial.SETTING: Department of Pediatrics, the Second Affiliated Hospital of Sun Yat-sen University; Biotechnology Research Center, Sun Yat-sen University.MATERIALS: The experiment had been done in the laboratories for Medical Research Center of the Second Affiliated Hospital, Sun Yat-sen University and Biotechnology Research Center, Sun Yat-sen University from May 2004 to January Lentiviral RNAi Expression System was purchased from Invitrogen, Co.,Ltd.; human VEGFR2 Mcb (PE) was purchased from R&D; CD31 immunohistochemistry kit was purchased from Boster, Co.,Ltd.; CD33-PE fluorescence labeled antibody was purchased from BD, Co.,Ltd.transiently and expression clone (Lenti6/shVEGFR2) was constructed, then cotransfected with ViraPowerTM Packaging Mix pU6/shVEGFR2 entry clone and transducting with Lenti6/shVEGFR2 expression clone, the effect on the development of intravenous xenograft leukemia mouse model, the distribution of microvessels in mouse bone marrow was observed after leukemia model mouse injected with recombinant lentivirus (group B); leukemia model mouse injected with recombinant lentivirus and endothelial cell (group C); leukemia model mouse injected with endothelial cell (group D). Through detecting changes of CD33 positive cells and microvessel density (MVD) in bone marrow, observing peripheral blood cell (PBC)smear and slice of liver, spleen, the effect of Lenti6/shVEGFR2 recombinant lentivirus on mouse leukemia was evaluated.mediated with lentivirus on VEGFNEGFR2 paracrine and autocrine loops in leukemia mouse.effective in inhibiting HL60 cell. pU6/shVEGFR2 entry clone constructed according to it had cell inhibitory rate as high as after transfection of pU6/shVEGFR2 entry clone and transduction of Lenti6/shVEGFR2 expression clone: 48 hours after transfection of pU6/shVEGFR2 entry clone and transduction of Lenti6/shVEGFR2 expression clone, the cell growth inhibitive rates were similar. However, the cell growth inhibitive rate of entry clone descended rapidly after 48 hours (P＜0.01); which of expression clone changed slowly, reaching the peak at 96 hours, dropped slightly, having no significance mouse: The amount of HL60 cells in bone marrow of groups A, B and C detected with flow cytometry were (25.8%±4.9)%, (14.3%±5.1)%, (8.4±2.6)%, respectively (P＜0.05); MVD in group C was obviously less than that in group D (P＜0.05); The amount of HL60 cells in leukemia model mouse injected with recombinant lentivirus and endothelial cell was the lowest as compared with the other groups.

Objective　To investigate the prognosis in risk of ventricular arrhythmia in coronary heart disease with heart rate variability (H RV), left ventricular ejection fraction (LVEF) and other clinical background dat a. Methods　A total of 81 patients were divided into ventricular premature beats (VPBs)≥30/h group and VPBs<30/h group. Their LVEF, HRV and cli nical data were studied and analyzed. Results　The age and blood pressure between 2 groups had no significant difference. LVEF, standard deviati on of all normal RR intervals (SDNN), SD of the average of NN interval (SDANN) a nd HRV triangular index (HRVI) were significant less in VPBs≥30/h group than in VPBs<30/h group (43.29±15.38 vs 67.33±11.47，P＜0.01；90.05±22.2 9 vs 117.90±30.32，P＜0.05；77.43±17.78 vs 105.69±28.79，P＜0.05 ；24.54±8.70 vs 32.70±10.87，P＜0.05, respectively). Incidence of myo cardial infarction (MI) was larger in VPBs≥30/h group than VPBs<30/h group. LVE F was the independent predictable factor in risk of ventricular arrhythmia with multinomial regression logistic analysis（B=0.119, P=0.032）. Co nclusion 　Our findings indicate that LVEF is an independent predictable factor i n risk of ventricular arrhythmia in coronary heart disease. Although HRV and MI history can not be used to predict VPB, significant difference is found between 2 groups. High-risk patients could be selected successfully when these data are considered in combination.

Objective To study the relationship between mean platelet volume(MPV)and severity of acute pancreatitis.Methods MPV was examined by hematology analyzer.Relationship between MPV and the severity of acute pancreatitis and the effect of sandostatin on MPV in acute pancreatitis were analyzed.Results The MPV of severe acute pancrcatitis was significantly higher than that of controls(P＜0.05),whereas the MPV of mild acute pancreafitis was not(P＞0.05).The APACHE Ⅱ Score and MPV of GH group was significantly lower than that of SS group and control(P＜0.05).Conclusion The MPV could reflect the severity of acute pancreatitis,evaluate the prognosis of acute pancreatitis.

Objective To observe the effects of low-frequency pulsed electromagnetic fields (LFPEMFs) on microcirculation angiogenesis in the hindlimbs of diabetic rats with acute ischemia. Methods Models of acute hindlimb ischemia were established in 60 male Sprague-Dawley diabetic rats. The diabetes model was established using 60 mg/kg intraperitoneal injections of streptozotocin (STZ). Fasting blood glucose levels were greater than 300 mg/dL. The rats were randomly divided into experimental and control groups. The rats in the experimental group were exposed to low-frequency pulsed electromagnetic fields for 2 hours each day, while the control group was not given any treatment. Laser-Doppler perfusion was used to measure blood flow in the ischemic hindlimb on days 0,7, 14 and 28 after the operation. The immunofluorescence of rat endothelial cell antigen-1 ( RECA-1) was used to evaluate the changes in angiogenesis. The levels of vascular endothelial growth factor (VEGF) and fibroblast growth factor-2 (FGF-2) were determined by both Western blotting and ELISA, and VEGFR2 and FGFR1 levels in the ischemic skeletal muscle were determined by Western blotting on days 7, 14 and 28 after the operation. Results The average perfusion ratio was significantly greater in the experimental group at days 14 and 28 compared with the control group. RECA-1 density in the tissues had increased significantly in the experimental group at the 14th and 28th day. The same was observed for FGF-2 and its receptor, but there was no significant difference for VEGF or its receptor in either group. Conclusions LFPMEFs can promote angiogenesis in acute hindlimb ischemia of diabetic rats by up-regulating FGF-2. This suggests that LFPMEFs may be useful for preventing and treating lower limb ischemia in diabetic humans.

Objective:To study the effect of IL-15 eukaryotic expression plasmid on the humoral immune responses to HBV surface antigen protein vaccine.Methods:Had constructed two of IL-15 eukaryotic expression plasmides.One is the eukaryotic expression plasmid of the IL-15 whole sequence,the other is the chimeric eukaryotic expression plasmid of the IL-2 signal peptide and IL-15 mature peptide sequence(IL-2s-15).The bioactivities of the expression products of the two plasmids were identified by CTLL-2 proliferation assay.Results:After IL-15 plasmid co-immunized with HBsAg,the titer of anti-HBsIgG was much higher than that of control( P 0.05),however,the anti-HBsIgG2a/IgG1 ratio was higher than that of control.Conclusion:IL-15 expression plasmids effect differently on the immune responses induced by protein vaccine.

AIM: To observe the effects of valproic acid(VPA) and HA14-1 on Bcl-2 overexpressed leukemia cells in vitro and in vivo.METHODS:(1) Cells were divided into control group,HA14-1 group,VPA group and HA14-1+VPA group.The apoptotic rate,the mean fluorescent index(MFI) of Bcl-2,the levels of caspase 3,8 and 9 were detected with FCM.(2) 24 h after transplantation with BALL-1,the NOD/SCID mice were divided into 4 groups as(1),then the survival time was compared.The expression of CD19 in the peripheral blood cells,bone marrows,livers,spleens and lungs of each group was detected.RESULTS:(1) The apoptotic rate in HA14-1+VPA group was(76.5?6.9)%,this was significantly elevated compared to the data in other groups(P

Objective To investigate the relationship between bone mineral density (BMD) and the changes of serum adiponectin (APN) level in elderly men.Methods A total of 240 elderly men was enrolled in this study.Measurement of BMD of lumbar spine with dual energy X-ray absorptiometry was performed.All subjects were divided into three groups (normal,osteopenia,and osteoporosis) according to the T value of BMD.Serum APN level was tested using enzyme-linked immunosorbent assay (ELISA),meanwhile were recorded as bone lurnover markers.Results (1) The level of serum APN in osteopenia group was lower than in normal group,the level of serum APN in osteoporosis group was significantly lower than in osteopenia group,while the level of serum APN in osteoporosis group was significantly lower than in normal group (all P ＜ 0.05).(2) Serum APN was positively correlated with T value of BMD (r =0.475,P ＜0.01).(3) The stepwise multiple linear regression analysis showed serum APN,25-hydroxyvitamin D,β-isomerized carboxyterminal propeptide (β-CTX),and osteocalcin could enter into the equation.Conclusions Serum APN might play an important role in pathogenesis of osteoporosis in elderly men.

Object To optimize the process for the extraction of the original recipe used in the treatment of eczema to give a new ECZEMA SPRAY dosage form Methods The extraction process was studied by orthogonal experimental design as guided by determining the content of paeonol and baicalin in the extract Results The optimal extraction process was to reflux the original recipe with 80% ethanol twice at a bath temperature of about 90 ℃ for 1 5 and 1 0 h respectively The amount of ethanol used for each extraction was 10 and 8 times of the original recipe respectively Conclusion The above extraction process gave the most rational and satisfactory results

This animal experiment was aimed at the questions whether high glucose concentration inhibits insulin secretion (glucose toxicity, GT) of beta-cell of islets from SHR and Wistar-Kyoto (WKY) rat and whether adrenomedullin (AM) enhances GT. Ten 6-week-old SHRs (test group) and ten 10-week-old Wistar-Kyoto rats (WKY) (control group) were selected. RAMI-1640 medium containing 5.6 mM glucose (normal glucose group) and 20 mM glucose (high glucose group) were applied. Various concentrations of AM (0, 10(-8), 10(-7), 10(-6) M) and RPMI-1640 medium containing high glucose were mixed, respectively. The isolated islets from rats were put into 12-well plates (90 islets/well). The islets were incubated in RAMI-1640 medium containing normal or high glucose for one hour. Then the supernatants from both incubations were determined by RIA for insulin. In SHR group, the insulin concentration in supernatants gained from high glucose group without AM was lower than that from normal glucose group (19.9+/-6.6 vs 60.9+/-33.6 mU/L, P<0.05). With the increment of the concentration of AM, insulin concentration in supernatants from islets incubated in high glucose and various concentrations of AM tended to be low further (19.9+/-6.6 vs 22.2+/-8.0 vs 21.5+/-5.6 vs 17.9+/-3.6 mU/L). The changing tendency in control group was the same as in SHR group. When the islets were incubated in normal glucose and high glucose medium, the insulin concentration in supernatant significantly decreased in SHR group compared with that in control group (P<0.01). The insulin secretion was inhibited by high glucose in beta-cell of islets from SHR and WKY. The results suggest GT to beta-cell of islets from SHR and WKY. AM tended to inhibit insulin secretion in a dose-dependent manner in beta-cell of islets from SHR and WKY. The inhibition of insulin secretion caused by high glucose in beta-cell of islets from SHR was more remarkable than from WKY. This may be related to secretion dysfunction in beta-cell of islets from SHR.
Adrenomedullin ; Animals ; Blood Glucose ; metabolism ; Cells, Cultured ; Female ; Glucose ; pharmacology ; Hyperglycemia ; blood ; Hypertension ; metabolism ; pathology ; Insulin ; secretion ; Insulin Resistance ; Islets of Langerhans ; drug effects ; pathology ; secretion ; Male ; Peptides ; pharmacology ; Rats ; Rats, Inbred SHR ; Rats, Inbred WKY

OBJECTIVETo study the chemical constituents from Aeschynanthus longicaullis.

METHODThe column chromatographic techniques were applied to isolate the constituents. The spectroscopie methods were used of EI-MS and NMR to identify the structures of the separated compounds.

RESULTSeven compounds were isolated from the ethyl acetate extract of A. longicaullis, whose structures were elucidated as cryptomeridiol (1), 4 (15) -eudesmene-1beta, 6alpha-diol (2), 2,5-bornanediol (3), isovanillic acid (4), vanillic acid (5), stigmast-5 (6) , 22 (23)-diene-3beta-ol (6) and beta-sitosterol (7).

CONCLUSIONCompounds 1-5 were isolated for the first time from Gesneriaceae and 6-7 were isolated for the first time from this plant.