To assess the effects of low dose aprotinin on platelets during cardiopulmonary bypass(CPB), 45 Patients, aged 20 to 45 years, ASA grade Ⅱ to Ⅲ,undcrgoing elective operation of cardiac valve replacement under fentanyl-enflurane-atracurium anesthesia, were randomly allocated to receiving aprotinin 2 ?106 KIU (group I,n=23 ) or equivalent volume of normal saline as control (group Ⅱ,n= 22) res12ectively,at the same time as CPB began to work. The blood samples were collected after induction of anes- thesia. 5 mins prior to CPB. 5, 15, 30 and 60 mins following CPB,and immedialely before withdrawal of CPB.to measure such 1parameters of platelet as level of cytoplasmic free Ca2+ (CFCa2+),activities of phosphollpase A2 (PLA2) and cyclouoxygenase (PCO) in membrane,aggregation rate (AGR),adhesion rate (ADR ) and count Individually. The lotal blood volume of postoperative tho- racic drainage solution (TBV ) and the re(]uircd blood volume of transfusioll (RBVT) were recorded. The results ahoys(l that dur lug CP14,the levels of CFCa2+ and activitics of PLA2 and PCO increased significantly,and the AGR,ADR and count of plstclet decreased markedly in both groups (P

The study was designed to evaluate the in vitro effects of aprotinin on platelets functions and arachidonic acid metabolism, Venous blood was drawn from healthy volunteers, and aprotinin was added to the blood at three different dose of aprotinin:50KIU/ml,150KIU/ml(low-dose)and 300KIU/ml (high-dose), respectively. Results demonstrated that aprotinin inhibited significantly the rates of platelet adhesion and aggregation induced by ADP or thrombin. Both the inhibitions were reversible and there was not significantly difference between high-dose and low-dose. The ADP-induced activities ofphospholipase A_2 (PLA_2) and cyclo-oxygenase(CLO)of platelet membrane were significantly suppressed by prophylactic aprotinin,but were not by therapeutic aprotinin. It is suggested that administration of low-dose aprotinin during CPB is reasonable,especially gaved just before the start of CPB.

Objective: To explore the effect of resveratrol (Res) on angiogenesis of human umbilical vein endothelial cells (HUVEC) with the possible mechanisms in vitro.
Methods: The HUVECs were cultured in 6 groups.①Control group, HUVEC were cultured with high glucose in DMEM,②Res group, the cell were cultured with Res at different concentrations,③Res with PI3K blocker LY294002 (Res+Blocker 1) group, ④Blocker 1 group, HUVEC were cultured with LY294002 alone, ⑤Res with eNOS blocker L-NAME (Res +Blocker 2) group and ⑥Blocker 2 group. The effect of Res on HUVEC proliferation was detected by CCK-8 kit, the protein expressions of p-Akt, p-eNOS were examined by Westin blot analysis, nitric oxide (NO) level was measured by nitrate reduction method and the endothelial cell migration was assayed by transwell chamber method.
Results: ① Compared with Control group, HUVEC proliferation increased in Res (1, 5μmol/L ) group, P<0.01, the proliferation in Res (5μmol/L) group was higher than those in Res (0.2, 10, 20μmol/L) group, P<0.01, while Res (20 μmol/L) group could inhibit the proliferation P<0.01. ②Compared with Control group, Res (5μmol/L) group had the higher protein expressions of p-Akt, p-eNOS, P<0.05-0.01, higher NO level, P<0.05.③Compared with Res group, Res+Blocker 1 group had lower expressions of p-Akt, p-eNOS, P<0.01, lower NO, P<0.05; the expressions of p-Akt, p-eNOS and NO level were similar between Res+Blocker 1 group and Blocker 1 group, all P>0.05.④Compared with Control group, the cell migration and tubing formation were higher in Res (5μmol/L) group, P<0.01;compared with Res group, the cell migration and tubing formation were lower in Res+Block2 group, P<0.01.
Conclusion: Res could up-regulate NO level via activating PI3-K/Akt/eNOS signaling and therefore, improving the proliferation, migration and tubing formation of HUVEC in vitro.

Objective: To explore the effect of atorvastatin (Atv) on high glucose-induced oxidative stress injury in human umbilical vein endothelial cells (HUVECs) by SIRT1/NADPH oxidase pathway with the possible mechanisms.
Methods: HUVECs were cultured in low glucose medium and then divided into 6 experimental groups:①Normal group,②Osmotic pressure control group,③High glucose (HG) group,④HG+Atv (0.1, 1.0, 10.0)μmol/L group,⑤HG+sirtinol (SIRT1 inhibitor) group,⑥HG+apocynin (NOX4 inhibitor) group, and HUVECs were further cultured for 24 hours. The cell proliferation was examined by CCK-8 kit, ROS level was detected by lfow cytometry method, protein expressions of SIRT1 and NOX4 were measured by Western blot analysis.
Results: ① Compared with Normal group, HG group had decreased HUVECs proliferation, Atv improved the HG inhibited proliferation in a does dependent manner. ② HG group had the higher level of ROS, increased NOX4 protein expression and decreased SIRT1 protein expression. ③ In HG condition, Atv up-regulated SIRT1 expression and down-regulated ROS and NOX4 expressions in a does dependent manner.④In HG condition, sirtinol decreased SIRT1 expression, increased NOX4 expression, and apocynin decreased NOX4 expression, while it had no inlfuence on SIRT1 expression.
Conclusion: Atorvastatin could resist HG-induced oxidative stress injury in HUVECs, which might be related to up-regulated SIRT1 expression, and SIRTI plays the role in NADPH oxidase at upstream.

Traditional research ideas of miRNA-target gene-biological function have ignored the contact between the target genes and signaling pathways involved , making the integrity and relevance of miRNA regulatory mechanisms not be fully elucidated .Integrated with systematic and relevant way of thinking , summarization and analysis for the luciferase reporter assay validated miR-205 target genes and their related signaling pathways will pave the way for new research area for miR-205 , and , it will be helpful for breaking through the status quo and exploring the novel research areas for miRNA .

Objectiye To optimize the depth of the microchannel and the time point for sperm collection,and improve the efficiency of sperm screening on a microfluidic device. Methods Microchannels with four different depths of 25, 50, 100 and 200 μm were tested. Mice sperm were added to the inlet of the microchannel. The relative quantity and motility of sperm in the outlet were recorded at different collection times, i.e. ,5, 15, 30 and 60 min. Statistical method one-way ANOVA and appropriate post-hoc testing were applied to analyze differences between different groups, and further to select the best-fit depth of the microchannel and the time point for collection. Results In microchannels with depths of 25, 50, 100 and 200 μm, the sperm motilities measured in each outlet were (85.4 ± 2.3)%, (85.8 ± 5.8)%,( 87. 2 ± 2. 8 ) %, (76. 5 ± 2. 8 ) % respectively with statistical significance ( F = 5.8, P ＜ 0. 05 ). No obvious differences were found among 25-100 μm channels, however the motility dramatically decreased in the 200 μm group. The relative sperm quantities were (5.2 ±2.0)%, (7.2 ±2.5)%,(12.3 ±2.0)%,(7. 7 ± 1.1 ) % respectively with statistical significance ( F = 6. 9, P ＜ 0. 05), which increased with channel depth from 25 to 100 μm,while it decreased in the 200 μm channel Taking 2 indexes into account, 100 μmwas the most fit channel depth for sperm motility screening. The sperm motility in the outlet gradually decreased with time. At the time points of 5, 15, 30 and 60 min after adding sperm, the sperm motilities were (99. 6 ±0. 7)%, (87.2 ±2. 8)%, (79. 3 ±2. 2)% and (62. 6 ±8.0)% respectively with statistical significance ( F = 37. 3, P ＜ 0. 01 ). Yet the relative quantities of sperm in the outlet increased almost three times in this process. At the time points mentioned above, the relative quantities of sperm were (5.8±1.1)%, (10.6 ± 0.9)%, (12.1 ± 1.7)%, (17.9 ± 3.4)% respectively with statistical significance ( F = 17.8, P ＜ 0. 01 ). Thus 15-30 min was the ideal screening time. Conclusion An effective microdevice for sperm screening with optimized depth and collection time period is developed,which may contribute significantly for the screening of healthy sperm on microfluidic chips.

BACKGROUND:Currently, antisense oligonucleotides (AS-ODN) have a good prospect in gene therapy, but AS-ODN with smal molecular weight cannot easily enter into the cels, which is susceptible to nuclease degradation. Therefore, there is stil a lack of fundamental understanding about how to improve their transfection efficiency, and target-based transferring. OBJECTIVE:To investigate whether a weak cationic and phosphorylcholine-containing diblock copolymer (MPC30-DEA70) can act as a carrier system to deliver a chemicaly synthesized transforming growth factor-β1 (TGF-β1) AS-ODN into myocardial cels. METHODS: MPC30-DEA70 was compounded with TGF-β1 AS-ODN at various N/P ratios and the MPC30-DEA70/TGF-β1 AS-ODN complexes were characterized by DNA electrophoresis. MTT assay was used to observe the biocompatibility. Confocal laser scanning microscope was used to observe the distribution and location of MPC30- DEA70/TGF-β1 AS-ODN in cells. Flow cytometry was used to detect the transfection efficiency and fluorescence intensity of MPC30-DEA70/TGF-β1 AS-ODN in cells. Western blot and RT-PCR methods were employed to measure the expression of TGF-β1 in cells. RESULTS AND CONCLUSION: Cell growth inhibition showed that the MPC30-DEA70 had low cytotoxicity to myocardial cells within the effective transfection dosage range (< 20 mg/L). Data from the flow cytometry test indicated a clear trend of increasing transfection efficiency with the increasing of N/P ratios. At high N/P ratios, the expression levels of TGF-β1 mRNA and protein in myocardial cells were significantly lower. This study shows that MPC30-DEA70 can work as an effective transgenic vector in myocardial cells. TGF-β1 AS-ODN can silence the expression of TGF-β1 gene efficiently and specially, and may antagonize TGF-β1-mediated biological function.

AIM: To detect the effects of resveratrol (RSV) on the expression of microRNA-21 (miR-21) in primarily cultured neonatal rat atrial myocytes with electric remodeling induced by rapid electrical stimulation (RES).Furthermore, to find out the possible mechanism of miR-21 regulating electrical remodeling.METHODS: The neonatal rat atrial myocytes were isolated by double-enzyme (trypsin and collagenase I) digestion and differential adhesion method.The atrial fibrillation (AF) model was induced by RES.Atrial myocytes were randomly divided into 4 groups: control group, RSV group, RES group, and RSV+RES group.To further detect whether RSV regulated electric remodeling by miR-21, except the 4 groups, we add miR-21 over-expression group and miR-21 inhibitor group: RES+negative control (NC) group, RES+miR-21 mimics group, RES+miR-21 mimics+RSV group, RES+miR-21 inhibitor group, and RES+miR-21 inhibitor+RSV group.The optimal concentration and pretreatment time of resveratrol were determined by CCK-8 assay.The expression of miR-21 and the mRNA expression of L-type calcium channels CACNA1C and CACNB2 in atrial myocytes were detected by qPCR.The protein expression of L-type calcium channels Cav1.2 and Cavβ2 in the atrial myocytes was analyzed by Western blot.RESULTS: The expression of miR-21 in RES group was significantly increased compared with control group, while preconditioning with RSV decreased the expression of miR-21.Compared with RES+miR-21 mimics group, the expression of miR-21 in RES+miR-21 mimics+RSV group was significantly decreased.Meanwhile, the mRNA expression of CACNA1C and CACNB2, and the protein levels of Cav1.2 and Cavβ2 were increased (P<0.05).Compared with RES group, the expression of miR-21 in RES+miR-21 inhibitor group and RES+miR-21 inhibitor+RSV group was decreased, while the mRNA expression of CACNA1C and CACNB2, and the protein levels of Cav1.2 and Cavβ2 were increased.However, no difference of the expression of miR-21, the mRNA expression of CACNA1C and CACNB2, and the protein levels of Cav1.2 and Cavβ2 among RSV+RES, RES+miR-21 inhibitor and RES+miR-21 inhibitor+RSV groups was observed (P<0.05).CONCLUSION: In AF model induced by RES, RSV may reduce electric remodeling by inhibiting the expression of miR-21 and regulating the downstream target genes.

AIM:To investigate the effects of Akt1 gene transfection into myocardium after ischemia-reperfusion (I/R) on mitochondrial permeability transition. METHODS:Forty adult male SD rats were divided randomly into five groups with 8 rats each:control group,I/R group,Ad-gene group,Ad-blank group and Ad-inhibitor group. The rats in Ad-gene group were injected with 30 μL Lipofectamine 2000 solution including Akt1 gene to the myocardium 48 h before ischemia while those in control group and I/R group were injected with PBS of the same volume. Rats in Ad-blank group were injected with Lipofectamine 2000 of the same volume into myocardium. In Ad-inhibitor group 30 μL Lipofectamine 2000 and gene complexes with LY294002 were injected. Hemodynamics,apoptotic index,the concentrations of lactate dehydrogenase,creatine kinase,the expression of Akt1,cytosolic,mitochondrial cytochrome C and MPT were also measured. RESULTS:The lowest level of Akt1 protein expression was observed in control group. The protein expression of Akt1 in Ad-gene group was higher than that in I/R group,Ad-blank group and Ad-inhibitor group. The AI,LDH and CK in Ad-gene group were significantly lower than those in other groups except control group. Transfection of Akt1 markedly reduced the loss of mitochondrial cytochome C after I/R injury. Ad-gene transfection led to a significant increase in absorbance at 540 nm compared to I/R group,Ad - black group and Ad-inhibitor group (P<0.05). CONCLUSION:Akt1 gene prevents myocardial apoptosis after I/R injury. Akt1 gene also inhibits the opening of mitochondria permeability transition and protects mitochondrial functions of myocardium in I/R injury.

Objective To identify the prevalence and profile of psychological difficulties in a sample of women seeking the breast reconstruction, and to study the effect of the reconstructive surgery on their psychology. Methods Before and after the breast reconstruction, 50 breast-loss women were tested with the self-rating anxiety scale (SAS), the self-rating depression scale (SDS), the self-esteem scale (SES),the self- rating body dysmorphic disorder scale (SBDDS), and the Eysenck personality questionnaire (EPQ), respectively. The proportion of the abnormally psychological patients was counted, their personality characters were analyzed, and the postoperative psychological changes were obverved. Results It was found that the 64 percent of the breast-loss patients were in anxiety, 30 percent were in depression, and 18 percent were in body dysmorphic disorder. The characteristics of their personality were more extravert, sociable and easily emotional agitation. The scores in SES of the preoperative patients were 27.46±8.95, and those of the postoperative patients were 33.05±6.12 (P＜0.05). The scores in SBDDS of the preoperative patients were 25.74±13.23, and those of the postoperative patients were 18.22±8.08 (P＜0.05).Conclusion The proportion of the anxiety, depression and BDD is high in the breast-loss women, and the postoperative psychology of esteem and body dysmorphic disorder is improved effectively.