Objective To determine the changes of L-type calcium currents(I Ca-L ) of rabbit ischemic myocytes in peri-infarcted zone.Methods Rabbit AMI models were made by ligation of branches of left coronary artery. After 1 week and 2 months, single pericardial myocytes were isolated enzymatically from ischemic zone adjacent to infarcted area and the normal control cells from similar regions in normal rabbit hearts. I Ca-L were recorded by using patch clamp techniques in the whole cell configuration. Results Peak of I Ca-L density of ischemic cells from the peri-infarcted zone was significantly reduced in AMI onc week later ( 3 52 0 93 pA/pF, n=6) compared to control cells ( 5 58 1 53pA/pF,n=10), P0 05 The steady-state inactivation curves were shifted to the hyperpolarizing direction in ischemic cells,the half-maximal voltage dependence of inactivation(V 1/2 ) in ischemic cells was 25 9 7 0 mV in AMI 1W , 21 3 5 6mV in AMI 2M and 13 1 4 2mV in the control cells(compared with control group,P

Aim: To set a suitable route for efficient expression and purification of recombinant hyaluronan syn-thase from Pasteurella multocida (rPmH AS). Methods: Coding sequences were cloned and expressed in Esche-richia coli strain of BL-21( DE3). Then rPmHAS was purified through a simple Ni-affinity chromatography and identified by hyaluronic acid binding protein ( HABP) based ELISA assay. Results: High yield expression of functional rPmHAS exceeding the highest production reported hitherto was achieved, and the purity of rPmHAS was as high as 90%. Conclusion: In this study, an optimized system of the expression, purification and identifica-tion route for large scale preparation of rPmHAS was established, which will be helpful for accelerating further study of PmHAS and facilitating fine researches on peculiar hyaluronic acid with special properties.

Taking human umbilical vein endothelial cells (HUVEC)as experimental model which can simulate tumor-derived vascular endothelial cells;the effects of CPU-XT-008;an amino sugar derivative of combretastatin A-4 (CA-4);on HUVEC proliferation;cell cycle distribution;tubulin polymerization and the key regulatory pro-tein of cell cycle were studied.The effect of CPU-XT-008 on the proliferation of HUVEC was determined by MTT assay.The cell cycle distribution caused by CPU-XT-008 was detected by flow cytometry.Immunofluorescence was used to detect apoptosis and tubulin polymerization.The expressions of Cyclin B1;Cdc2 and β-tubulin were detected by Western blotting.Results demonstrated that CPU-XT-008 could target tubulin and inhibit the poly-merization ofβ-tubulin;and it could lead to G2/M cell cycle arrest in HUVEC by up-regulating Cyclin B1 expres-sion and down-regulating Cdc2 and p-Cdc2 expression;which resulted in inhibiting the proliferation of HUVEC and inducing its apoptosis.

In this study, the effect of heparin-derived oligosaccharide (HDO) on platelet-derived growth factor (PDGF) induced vascular smooth muscle cells (VSMCs) proliferation and the related signal transduction mechanisms were investigated. MTT assays were used to measure VSMCs proliferation. Cell cycle distribution was analyzed by flow cytometry. The level of key regulatory proteins in PKC, MAPK and Akt/PI3K pathways were determined by RT-PCR, Western blot and immunocytochemical methods. Meanwhile, mRNA expressions of some proto-oncogenes were assayed by RT-PCR method. Our data showed that HDO (0.01, 0.1 and 1 μmol · L(-1)) inhibited 30 ng · mL(-1) PDGF-induced VSMCs proliferation in a dose-dependent manner, blocked the G1/S transition and inhibited the level of key regulatory proteins and some proto-oncogenes (P < 0.05). The results showed that HDO may decrease the key regulatory proteins expression, hence suppress the transcription of proto-oncogene and G1/S transition, finally inhibiting VSMCs proliferation.

OBJECTIVE:To explore the effect and mechanism of ulinastatin on blood coagulation dysfunction of ICU sepsis pa-tients. METHODS:64 ICU sepsis patients were randomly divided into treatment and control groups,with 32 cases in each group. Control group received routine treatment,while treatment group was additionally given Ulinastatin injection on the basis of control group,100 000 u dissolved in 500 ml 15% Glucose injection or Sodium chloride injection intraveously,3 times/d,for consecutive 7 days. The mechanical ventilation time,ICU length of stay and survival rate within 30 d were analyzed statistically in 2 groups. The platelet count (PLT),prothrombin time (PT),activated partial thromboplastin time (APTT),fibrinogen (FIB),D-dimer (D-D), white blood cell count (WBC) and IL-6 were detected before treatment and on first,third and seventh day after treatment. RE-SULTS:After treatment,mechanical ventilation and ICU length of stay in treatment group were significantly shorter than in control group,and survival rate was significantly higher than control group,with statistical significance (P<0.05). The peripheral blood WBC and IL-6 level of treatment group were significantly lower than those of control group,with statistical significance(P<0.05). There was a significant difference in blood coagulation indicators between treatment group after 7 days of treatment and before treat-ment,control group after treatment(P<0.05). The blood coagulation indicators recovered to normal level after 7 days of treatment. CONCLUSIONS:Ulinastatin can improve blood coagulation of ICU sepsis patients by a mechanism of inhibiting the release of in-flammatory cytokines and corresponding blood coagulation factor function.

Objective To explore the role of cell proliferation and apoptosis, and the expression significance of involved protein as Bcl-2(B-cell lymphoma/leukemia-2) and Bax (Bcl-associate X protein) in the developing small intestine of human fetus. Methods The expression product of Bcl-2 and Bax was investigated with immunohistochemical methods in the 2nd, 3rd and 4th months of gestation respectively. Results In the second, the third and the fourth month of gestation, the Bcl-2 immunoreactive positive signals were found in the ganglion cells of intermuscular and submucous nerve plexus. Bax positive cells were observed in the cytoplasm of the simple columnar epithelium cells of the small intestinal mucous layer.Conclusion Bcl-2 and Bax proteins regulate the developing small intestine of human fetus.

Objective To observe the protective effect of?-asarone,the active components from Rhizoma Acori Tarinowii,on human umbilical venous endothelial cell strain ECV304 injury induced by oxidized low-density lipoprotein(ox-LDL)and to investigate its influence on proliferation of vascular smooth muscle cells(VSMC). Methods The well-grown ECV304 cells were randomized into 5 groups:normal group,control group,high-, middle-and low-dose?-asarone groups.Except the normal group,ECV in the other groups were co-cultured with ox-LDL 25?g/mL,and the?-asarone groups were additionally with?-asarone 30,15 and 7.5?g/mL added respectively.Flow cytometry was used to detect the cell apoptotic rate and the mitochondrial membrane potential (MMP)of ECV304 as well as the proliferation index and the DNA content in different cell phases of VSMC. Results In the model group,the apoptotic rate of ECV was increased,MMP was decreased,and VSMC proliferation was promoted(P

0.05, while the differences between 2 groups in cooling effects on the points of cooling 2.5 hours and 4.0 hours were significant, F

Objective To investigate the clinical and pathologic features of small gastrointestinal stromal tumors (small GISTs,d ＜ 2.0 cm).Methods Medical records of 95 patients undergoing surgery (endoscopic surgery,thoracoscopic/laparoscropic surgery and open surgery)and diagnosed as having GISTs by pathology and immunohistochemistry in Nanfang hospital from October 2003 to June 2014 were retrospectively analyzed.Based on clinical and pathological results,correlation analyses between risk factors for endoscopic ultrasonography(EUS) and Mitotic count(MI),clinicopathologic parameter and NIH risk classification were performed.Results Among 95 cases (104 lesions),88 were single,while 7 were multiple;81.7％ (85/104) small GISTs arose from stomach,including 87.1％ (74/85)in middle-upper stomach;5 cases (5.3％) presented calcification of different degrees,3 cases(3.2％) presented local necrosis and 2 cases (2.1％) with arrangement of epithelioid cells;88 cases (92.6％) were very low grade of NIH risk classification,6 cases (6.3％) were intermediate risk and 1 case(1.1％) was high risk.Positive rates of CD34 and CD117 were 95.8％ (91/95) and 96.8％ (92/95) respectively.The risk factors (border,mucosal surface,echo and heterogeneity) of EUS had no correlation with mitotic count(P＞0.05).The correlation analysis between clinicopathologic features and NIH risk classification revealed tumors more than 1.5 cm had a striking correlation with NIH risk classification (P＜ 0.05).Conclusion Most small GISTs,single or multiple,located at middle-upper stomach,were of very low or low risk,and have a favorable prognosis.But it has worse biological behavior and a higher grade risk when the diameter is more than 1.5 cm,intervention should be recommended.

Objective Three parameters include circumferential strain (CS),circumferential strain rate (CSr) and stiffness parameter (β) were measured to evaluate the elasticity of carotid artery using two-dimensional(2D) strain imaging and evaluate its clinical value in patients with type 2 diabetes mellitus (T2DM).Methods 60 patients with T2DM were collected and divided into carotid intima-midia thickness (CIMT) thickeness group(34 cases,1.0 mm＜CIMT≤1.2 mm) and plaque group(26 case,CIMT＞ 1.2 mm).33 normal people were supplied as control group.The systolic global peak CS and CSr of the carotid artery were obtained from short-axis view of the common carotid artery and the stiffness parameters β was measured using 2D strain imaging.All the parameters were compared among the three groups.Results The CS and CSr were decreased and β was increased in patients with T2DM (P ＜ 0.01),The CS and CSr obtained from the plaque group were less than that of the CIMT thickeness group(P ＜0.05),but the β was higher than that of the CIMT thickeness group (P ＜0.01).The CS and β were correlated significantly with CIMT respectively (r =-0.79,P ＜ 0.01 ; r =0.72,P ＜ 0.01).Conclusions 2D strain imaging is a technique for early assessing the carotid artery elasticity in patients with T2DM,the thicker the CIMT is,the smaller the deformation is and the larger the β is.