Objective To screen the active cardiovascular components from Rhizoma Polygonati and to observe the effect on myocardial ischemia rat.Methods The active components from Rhizoma Polygonati were extracted by system solvent separation methods,and their cardiovascular effects on the rat models induced by subcutaneous injection of isoproterenol were tracked to find out the optimal active cardiovascular components.Results Among the extracts from Rhizoma Polygonati,the n-Butanol(EB)extract which consisted of glycosides and aglycons,was the best active cardiovascular component;aqueous extract which consisted of alkaloids,flavonoids,glycosides and aglycons,was the better component;the emulsion of Rhizoma Polygonati,which consisted of petroleum ether extract,chloroform extract and water-ethanol extract had less cardiovascular activity.Conclusions The parts with medium polarity from Rhizoma Polygonati have the best cardiovascular activities.

Objective To screen the proteins interact with cytoplasmic tail of NECL1.Methods The cytoplasmic tail sequence of human NECL1 gene was inserted into the "bait" vector pGBKT7 in frame.The recombinant pGBKT7-NECL1C was then transformed into yeast cells and followed by screening of human fetal brain cDNA library.Then the GST pull down assay was used to confirm the interactions between the proteins.Results Nine different sequences by sequencing the positive clones and five alternative proteins were obtained by the amino acid sequence blast.Using the GST pull down analysis,we confirmed that two of them can interact with the cytoplasmic tail of NECL1 in vitro.Conclusion The alternative proteins obtained with the yeast dual-system may interact with the cytoplasmic tail of NECL1.

Objective To study the effect of PcG member NSPc1 on proliferation of HeLa cells.Methods Using bioinfomatic analysis to design the siRNA sequence to knockdown NSPc1.Detecting the expression level of NSPc1 in HeLa cell line using semi-quantitative RT-PCR,Real-time PCR and Western blot after transfection of the designed siRNA.Transient transfecting pSUPER-NSPc1 into Hela cells and performing BrdU incorporation assay.Establishing NSPc1 stably knockdown cell line,comparing proliferation abilities with the control cells.Results(1)The designed siRNA did efficiently knockdown the expression of NSPc1;(2)Transient knockdown of NSPc1 could repress BrdU incorporation;(3) The established NSPc1-knockdown cell lines had a significantly lower proliferation rate than that of control cells.Conclusion The expression of NSPc1 is necessary for the normal proliferation of HeLa cells.The NSPc1 stably knockdown cell pool is a useful model for further study of pathway related to NSPc1.

Objective To express the human recombinant SUMO1 protein and prepare monoclonal antibody(mAb) against it.Methods The recombinant expression plasmid pET32a-HIS-SUMO1 was made and transformed into E.coli(BL21),then the recombinant fusion protein HIS-SUMO1 was expressed and purified.The BALB/c mice were immuned with pure protein HIS-SUMO1 as antigen.Monoclonal antibody against SUMO1 was prepared with standard hybridoma technology.The hybridoma cell lines were obtained by ELISA and Western blot screening procedure,the isotype of the mAbs were further identified by immune-double diffusion.Ascites were collected from one propagated hybridoma cell line and mAbs were purified by using the Kit of Millipore.The valence of mAb was detected by Western Blot.Results The recombinant protein HIS-SUMO1 is expressed and purified.Three hybfidmas producing antibodies against SUMO1 were obtained,the isotypes of three mAbs are IgG1,Western blot showed that the antibodies were specific for SUMO1.The antibody purified from the ascites has better specificity.Conclusion The SUMO1 mAb prepared by using recombinant SUMO1 protein as antigen can be used for detectingthe protein sumoylation.

Objective To investigate the clinical outcome of the feasible method of the free string type dorsalis pedis flap and anterior malleolus flap in the repairement of skin defects caused by penetrating wound of palm. Methods From May 2004 to July 2009, the free string-type dorsalis pedis flap and anterior malleolus flap were used to repair skin defects of 16 cases who sutained penetrating wounds of palm.Results All 32 flaps in the 16 cases were all survived. Follow-ups were done from 6 months to 2 years after operation. Both the appearances of the repaired palms and the functional recovery were satisfactory. The function assessment of the hand was excellent in 8 cases, good in 5 cases, fine in 2 cases and poor in 1 case.The eligible rate was 81.25%. Conclusion The anatomy of the dorsalis pedis flap and anterior malleolus flap is rare variant, which facilitate the dissection of the flaps during operation. The repairement of skin defects at two sides, both the palm and the back of hand, can be achieved via one operation. Therefore, the free string type dorsalis pedis flap and anterior malleolus flaps offer an ideal procedure to repair skin defects secondary to penetrating wound of palm.

Objective To investigate the outcome of the free double-skin paddle string-type composite fibular flap in the reconstruction of the combined defects of ulna and radium. Methods From June 2005 to July 2009, 5 cases with combined defects of ulna and radium were reconstructed using the free double-skin paddle string-type composite fibular flap. The length of fibular segment for the reconstruction of ulnar defect ranges from 4.5 to 7.5 cm. The length of fibular segment for the reconstruction of radial defect ranges from 5.5 to 7.0 cm. The size of the flap varies from 5.0 cm × 3.0 cm to 8.0 cm × 5.5 cm. At the 12 month follow-up, the function of reconstructed forearm was evaluated based upon Enneking scoring system.Results Ten flaps in the 5 cases all survived. The time for the transplanted fibula healed on the radium and ulna was 4-6 months. The 5 patients were followed up from 14 months to 2 years. The forearm rotation functions were excellent in 2 cases, good in 2 cases and poor in 1 case. The eligible rate was 80%. The average Enneking score was 24.8, which indicated an average of 81.3% recovery of limb function. Conclusion Bone graft with blood supply can ensure the activity of osteocytes, which facilitates the fracture union.Whilst, the procedure can reconstruct multi-location and multi-tissue defects in the forearm. Therefore, the double-skin paddle string-type composite fibular flap is an ideal alternative for the reconstruction of the combined defects of ulna and radium and the skin.

Objective To evaluate the relationship of clinical symptom to plasmic levels of D-dimer, activated factorⅦ (FⅦa) and tissue factor pathway inhibitor (TFPI)/X a in patients with urticaria. Methods A total of 27 patients with chronic urticaria (CU), 27 patients with acute urticaria (AU) and 26 normal human controls were included in this study. Symptom score was determined and disease course was surveyed in these patients. ELISA was used to detect the plasma levels of D-dimer, FⅦa and (TFPI)/Xa in patients and controls. The relation of clinical symptom and disease course to plasma levels of these parameters was assessed. Results In patients with AU and normal controls, the plasma level of D-dimer was 450.57± 242.13 ng/mL and 266.81±40.68 ng/mL, respectively, the level of FⅦa, 2.23± 0.74 ng/mL and 5.23±1.35 ng/mL, respectively, and the level of TFPI/Xa 0.87±0.13 nmol/L and 0.88 ~ 0.12 nmol/L, respectively. There was a significant difference in the level of both D-dimer and FⅦa (both P < 0.01 ), whereas no differ-ence was observed in that of TFPI/X a (P > 0.05) between patients with AU and normal controls. In addi-tion, increased level of D-dimer and decreased level of FⅦa were noticed in patients with CU compared with those in normal controls (593.80±294.04 ng/mL vs 266.81±40.68 ng/mL, 3.98±0.35 ng/mL vs 5.23± 1.35 ng/mL, both P < 0.01 ), but there was no significant difference in the plasma level of TFPI/Xa (0.87± 0.16 nmol/L vs 0.88±0.12 nmol/L, P > 0.05). Significant difference was observed in the plasma level of D-dimer and FⅦa between patients with AU and CU (450.57±242.13 ng/mL vs 593.80 ±294.04 ng/mL, P < 0.05; 2.23± 0.74 ng/mL vs 3.98± 0.35 ng/mL, P<0.01 ). The plasma level of D-dimer positively corre-lated to the symptom score of patients with CU and those with AU (r= 0.68, P< 0.01; r= 0.82, P< 0.01),but was independent of discase course (P> 0.05). Neither the level of FⅦa nor that of TFPI/Xa correlated to symptom score or disease course of patients (all P > 0.05). Conclusions There is an overactivation of coagulation cascade, consumption of blood coagulation factors and secondary fibrinolysis in patients with urticaria, suggesting that plasma D-dimer and FⅦa may be associated with the clinical symptoms of urticaria.

Objective To synthetize a novel MR molecular imaging probe named USPIO?PEG?tLyP?1,and to evaluate its value in detecting U87 cells by MR imaging. Methods USPIO?PEG?tLyP?1 was synthetized by conjugating USPIO?PEG with tLyP?1. Neuropilin?1 expression levels of glioma cell lines were detected by Western blot. The cytotoxicity of USPIO?PEG and USPIO?PEG?tLyP?1 were assessed by MTT colorimetric assay. The uptake efficiency of USPIO?PEG?tLyP?1 was measured by Prussian blue staining, transmission electron microscope and MR imaging in vitro. Results The novel MR molecular imaging probe was synthetized with an average diameter of 43.84 nm. U87 glioma cell line was screened as test subject for the highly expression of NRP?1(P<0.05). USPIO?PEG?tLyP?1 group showed much more intracellular blue particles than USPIO?PEG group after Prussian blue staining. After incubation,USPIO?PEG?tLyP?1 mainly existed in lysosme,endoplasmic reticulum and mitochondria. In vitro MRI showed that USPIO?PEG?tLyP?1 significantly enhanced the negative contrast effect compared with USPIO?PEG(P<0.01). Conclusion The decoration of tLyP?1 enhanced targeting ability of USPIO?PEG to glioma cells and MR molecular imaging can be a promising method for early diagnosis of gliomas.

AIM:TodetecthemoglobinA1c(HbA1c)andparametersofbloodglucosefluctuationinChinesenewlydiag-nosed type 2 diabetes mellitus (T2DM) patients, and further to specify the factors that were related to mean blood glucose (MBG) in this population.METHODS:Newly diagnosed T2DM patients (n=90) from 4 hospitals in Guangdong province were enrolled, and subjected to 3 d continuous glucose monitoring (CGM) after testing for HbA1c and other laboratory tests.Blood glucose data collected during CGM were used to calculate MBG and parameters of blood glucose fluctuation.RESULTS: Correlation analysis revealed that MBG was significantly related to all parameters of blood glucose fluctuation, HbA1c, fast plasma glucose ( FPG) and 2 h postprandial glucose (P<0.01), but not to sex, age or blood lipid profile.Further analysis utilizing step-wise general linear model showed that HbA1c, absolute means of daily difference ( MODD) , difference between maximal and minimal glucose ( DMMG) and FPG had the strongest relation to MBG.CONCLUSION: Factors affecting MBG of the newly diagnosed T2DMpatients in our country include HbA1c, FPG, DMMG and MODD, and thus it may be prone to misleading results that only HbA1c is applied to estimate MBG in this population.

Objective To discuss the correlation between aortic elasticity and coronary artery calcification by CT.Methods Totally 111 patients who were diagnosed of coronary artery disease underwent coronary artery CTA.The images were qualified for aortic elasticity measurement.All patients were divided into calcification negative group (n=43) and calcification positive group (n =68).The calcification positive group was further divided into light,medium,and serious groups according to their calcification scores.The ascending aortic images were reconstructed every 5 ％ R-R intervals.The cross-sectional areas and diameters of aortic in each R-R interval were measured automatically,then diameter variation rate (％ A0),aortic distensibility (A0D),aortic compliance (A0C) and aortic stiffness (A0SI) were calculated to evaluate aortic elasticity.Correlation between aortic elasticity and coronary artery calcification were analyzed.Results ％ A0,A0 D,A0C were lower and A0SI was higher in calcification positive group than those in calcification negative group (all P＜0.05).There was no significant differences in the four reference indexes of aortic elasticity among light,medium,and serious groups in calcification positive group (all P＞0.05).Correlation analysis demonstrated negative correlations between ％ A0,A0 D,A0 C and calcification scores,and a positive correlation between A0SI and calcification scores.Conclusion Aortic elasticity is correlated with coronary artery calcification,and the combination of them will be beneficial for the early diagnosis of coronary heart disease.