针对泉州地区杧果优良品种缺乏、成熟期较集中等问题,引进"贵妃杧"和"四季杧"在泉州地区试种.结果表明,"贵妃杧"长势较好,产量稳定,果实可食率高,纤维少,品质佳,具有较好的推广价值;"四季杧"植株具有速生快长的特性,周年开花结果且稳产丰产,果实品质佳,还可丰富杧果淡季销售市场,推广前景看好."贵妃杧"平均单果质量547.9 g,可食率82.3％,可溶性固形物含量15.3％,果肉细嫩爽口;"四季杧"正造果平均单果质量358.4 g,可食率77.3％,可溶性固形物含量17.5％,果汁较多;"四季杧"反季果平均单果质量313.4 g,可食率74.7％,可溶性固形物含量22.5％,果汁较少但甜度更高.这两个品种在株产、平均单果质量、可食率、纤维含量、果实品质等方面均优于本地杧果,适宜在泉州地区进行大面积推广种植.

为筛选适宜福建省龙眼园套种的绿肥品种.选取15个绿肥品种,观测其在龙眼园中的生物性状,覆盖率、攀援性、抗逆性和鲜草产量等.结果表明,爬地蓝、圆叶决明在长势、鲜草产量、抗逆性以及虫害发生情况等方面较其他绿肥品种表现优,其中爬地蓝为多年生,具根瘤,可多次割刈,更具应用价值.本研究为龙眼园套种绿肥的推广种植提供参考.

为研究黄蓝粘虫板在套种爬地蓝果园一天各时段不同高度下诱杀害虫的效果,以离地(爬地蓝匍匐层高度)0.5、1.0、1.5和2.0 m处悬挂黄色和蓝色粘虫板,测定黄蓝板诱集害虫的种类与数量.结果表明,不同悬挂高度黄板和蓝板对害虫诱杀效果不同,黄板在诱杀害虫的数量和种类上好于蓝板,以悬挂高度0.5 m诱杀效果最佳.一天各时段下,黄板对粉虱、实蝇引诱效果明显好于蓝板,蓝板对蓟马引诱效果优于黄板,表明粉虱、实蝇具有趋黄性,蓟马则具有趋蓝性.了解套种爬地蓝果园害虫一天各时段活动规律,可为生草果园虫害综合防治提供参考依据.

本文围绕龙眼果园生态化栽培,阐述豆科绿肥爬地蓝的生长特点及其在龙眼园间种的生态效应和栽培关键技术.

Chronic heart failure (CHF) may be associated with an energy deficit in cardiac muscle. As levo-carnitine (LC) is involved in the production of myocardial energy, it is hypothesized that LC supplementation may ameliorate CHF symptoms. This multicentre, randomized, double-blind, and placebo-controlled study included 265 patients with CHF. Patients were randomized to receive either LC or placebo, twice a day. Endpoints were measured after 7 days of treatment. Primary endpoint was a reduction of at least one NYHA class. Secondary endpoints were changes in 6-min walk distance (6-MWD) compared with baseline, either alone or in combination with NYHA class decrease, left ventricular ejection fraction, and NT-proBNP level, together with adverse events. The primary endpoint was reached in 60.9% of patients treated with LC, compared with only 44.7% of the placebo group ( P = 0.012). Among the secondary endpoints, 6-MWD, alone or in combination with NYHA class, improved significantly in the LC group compared with placebo ( P = 0.0497 and P = 0.003, respectively). l-Carnitine was well tolerated. The lowest baseline values of plasma-free carnitine were observed in patients with NYHA classes III and IV where the effect of LC supplementation was greatest ( P = 0.002). Treatment with LC significantly improved CHF symptoms in Chinese patients, probably by correcting a status of carnitine insufficiency.

Platelet adhesion and aggregation are critical for hemostasis and atherothrombosis. Interaction between the GPIb complex and VWF initiates platelet adhesion and contributes to complete vessel occlusion, particularly at sites of stenosis. However, no therapeutic targeting this pathway has been developed. Previously, we demonstrated in murine thrombosis models that Anfibatide, a snake venom-derived GPIbα antagonist, inhibited botrocetin-induced VWF-GPIbα binding and inhibited thrombus formation in vitro and in vivo, without significantly activating platelets or changing bleeding. We subsequently hypothesized that Anfibatide would be a safe and potent anti-thrombotic agent in humans. We tested Anfibatide with human platelets in vitro and conducted the first anti-GPIbα phase I clinical trial with 94 healthy volunteers. In vitro, Anfibatide inhibited ristocetin-induced and low dose thrombin-induced human platelet aggregation. Surface plasmon resonance studies showed that Anfibatide binds to GPIbα and blocks binding of both VWF A1 domain and thrombin. In ex vivo perfusion chambers, Anfibatide strongly inhibited human platelet adhesion, aggregation, and thrombus formation on a collagen-coated surface, especially at high shear flow conditions. Importantly, Anfibatide effectively dissolved the preformed thrombi in perfusion chambers. Anfibatide incubation with human whole blood did not significantly change coagulation parameters measured by thromboelastography. After treating volunteers with Anfibatide, no serious adverse events, premature discontinuations due to adverse events, or deaths occurred during the study. Anfibatide bound to 95% of GPIb and inhibited 90% of platelet aggregation without prolonging activated partial thromboplastin time, prothrombin time, thrombin time, or bleeding time. There was no bleeding tendency, no alteration of platelet count, and no detectable anti-Anfibatide antibodies. In conclusion, Anfibatide provides anti-thrombotic effects through inhibition of both GPIbα-VWF and GPIbα-thrombin interactions. These results demonstrate that Anfibatide is a safe and potent anti-platelet reagent with great potential for future anti-thrombotic therapy.

Background Platelet adhesion and subsequent aggregation at the site of vascular injury are critical for hemostasis and thrombosis. It has been well accepted that interaction between the GPIb complex and von Willebrand factor (VWF) plays a key role in initiation of platelet adhesion, particularly at high shear. Platelet surface integrin αIIbβ3, through interaction with fibrinogen or other ligands, then mediates platelet aggregation to form a stable hemostatic plug or thrombus. Recently, the indispensable role of the GPIb-VWF interaction in platelet aggregation at extremely high shear (e.g. > 10,000s-1; areas of stenosis following arteriosclerosis and/or thrombus growth) has been highlighted. Therefore, both the GPIb complex and αIIbβ3 are considered major targets for antithrombotic therapies. Interestingly, although several inhibitors of αIIbβ3 have been developed for antithrombotic therapies, no drug has been developed to target the GPIb complex even though there are limitations for anti-αIIbβ3 therapies. The GPIb complex is, therefore, an attractive target for anti-thrombotic therapy. Here, we evaluated the efficacy and safety in vitro and in vivo of Anfibatide, a novel GPIb antagonist, in mice and in a phase I clinical trial.Methods Anfibatide was purified from venom of the Agkistrodon acutus snake and its purity was analyzed by mass spectrometry. The effect of Anfibatide on murine platelet function was assessed by in vitro platelet aggregometry, ex vivo perfusion chamber, and two complementary in vivo intravital microscopy models. The effects of Anfibatide on human platelet aggregation and thrombus formation were studied in vitro, and thrombealastography (TEG) was also performed. Most importantly, we evaluated the safety and efficacy of Anfibatide on platelet function and coagulation in a total of 94 healthy human volunteers in a phase I clinical trial.Results MALDI-TOF mass spectrometry of Anfibatide showed only one peak and the mass to charge ratio is 29799.7. Anfibatide specifically inhibited ristocetin-induced human platelet aggregation. Interestingly, Anfibatide was not able to inhibit botrocetin-induced murine platelet aggregation in plate-rich plasma (PRP), suggesting that its binding site may differ from other snake venom-derived GPIb antagonists. We found Anfibatide did not affect ADP-, TRAP- or collagen-induced aggregation in PRP, suggesting its specificity to GPIb. In ex vivo perfusion, Anfibatide strongly inhibited murine and human platelet adhesion, aggregation, and thrombus formation on a collagen-coated surface at both high and low shear flow conditions although it is far more sensitive at high shear. Importantly, Anfibatide effectively dissolved the preformed thrombi when we continuously perfused Anfibatide-treated whole blood through perfusion chambers, demonstrating its potential as an anti-thrombotic therapy. In the mesenteric arteriole thrombosis model, Anfibatide strongly inhibited platelet adhesion, thrombus formation, and prevented vessel occlusion in response to FeCl3 injury (P<0.05). At sites of laser-injured cremaster arterioles, Anfibatide also dramatically inhibited platelet accumulation and thrombus growth. Anfibatide did not cause significant murine platelet activation in vitro and had no significant change in coagulation parameters in TEG when we treated human whole blood with Anfibatide, suggesting it had minimal side effects. In the phase I clinical trial, results showed that Anfibatide can occupy approximately 95% of GPIb and inhibit up to 90% of ristocetin specific platelet aggregation. The inhibitory effect was undetectable four hours after Anfibatide was withdrawn. There were no serious adverse events, or deaths that occurred during the study. Anfibatide did not significantly prolong bleeding time, activated partial thromboplastin time (APTT), prothrombin time (PT), or thrombin time (TT). There was also no spontaneous bleeding or bleeding from blood collection sites. Anfibatide did not significantly affect platelet count and no anti-Anfibatide antibodies were detected in the subjects, suggesting that Anfibatide is well-tolerated in healthy individuals.Conclusion These comprehensive studies in mice and human subjects and in the first clinical trial clearly demonstrated that Anfibatide is a safe and potent anti-platelet reagent with great potential for future anti-thrombotic therapy.Disclosures: Hou: Lee’s pharmaceutical holdings limited : Research Funding. Lei: Lee’s pharmaceutical holdings limited: Research Funding. Zhao: Lee’s pharmaceutical holdings limited: Research Funding. Shen: Lee’s pharmaceutical holdings limited: Research Funding. Zhou: Lee’s pharmaceutical holdings limited: Research Funding. Wang: Lee’s pharmaceutical holdings limited: Research Funding. Marshall: Lee’s pharmaceutical holdings limited: Research Funding. Ni: Lee’s pharmaceutical holdings limited: Research Funding.

Snake venoms contain various biologically active proteins including C-type lectins and -like proteins (CTLs), phospholipases, proteinases and metalloproteinases, that could target diverse cellular processes. Among these active components, CTLs have demonstrated antithrombotic effects through their platelet agglutination and coagulation activities. In our study, we have detected anti-angiogenesis activity from the venom of Deinagkistrodon acutus snake. The partial purified fraction of snake venom with anti-angiogenesis activity was isolated through a stepwise separation in anion-exchange and cation-exchange chromatography. At the same time, the monoclonal antibody was generated and used for further protein purification by affinity chromatography. This highly purified protein was named as ZK002. By MALDI-TOF mass spectrometry determination, the mass of ZK002 is about 30KD. By LC-MS/MS analysis, protein sequence of highly purified ZK002 was determined and compared with the Deinagkistrodon acutus snake protein database. ZK002 is a heterodimer of CTLs consisting of alpha and beta peptide chains. Using In vitro chicken embryo chorioallantoic membrane assay and human umbilical vein endothelial cell (HUVEC) tube formation assay, it showed that ZK002 exhibited an anti-angiogenesis effect. Moreover, ZK002 also demonstrated an anti-cancer effect as shown in the ApcMin/+ mouse model for colorectal cancer. Administration of ZK002 significantly suppressed the total number and the size of tumors and showed no observable adverse effect. To further elucidate the anti-angiogenesis functions of ZK002, we deduced the coding DNA sequences from the peptide sequences of the alpha- and beta-chains, then subcloned individual gene sequence into yeast and mammalian expression vectors. Results showed that the recombinant vectors express secreted peptides with the expected molecular weight. Transient transfection of the alpha- and beta-recombinant vectors individually or in combination inhibit tube formation in HUVEC cells, suggesting that both alpha- and beta-chains possess anti-angiogenesis activity. The success of cloning and expression of recombinant ZK002 would greatly facilitate the future development of ZK002 as anti-angiogenesis and anti-cancer drugs. (This work is supported by the Innovation and Technology Fund (UIM208) of Hong Kong Government) Citation Format: Shan Yu, Xiangrong Dai, William C.S. Tai, Roy C.Y. Choi, Chaofan Liang, Benjamin X.y. Li, Karl W.K. Tsim, Wendy W.L. Hsiao. Anti-angiogenesis and anti-cancer activities of ZK002, a C-type lectin-like protein isolated from snake venom. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 5077. doi:10.1158/1538-7445.AM2013-5077

Objective— To analyze the role of toll-like receptor 4 in modulating metabolism and endothelial function. Approach and Results— Type 2 diabetic mice with mutated toll-like receptor 4 (DWM) were protected from hyperglycemia and hypertension, despite an increased body weight. Isometric tension was measured in arterial rings with endothelium. Relaxations to acetylcholine were blunted in aortae and mesenteric arteries of Lepr db/db mice, but not in DWM mice; the endothelial NO synthase dimer/monomer ratio and endothelial NO synthase phosphorylation levels were higher in DWM preparations. These differences were abolished by apocynin. Contractions to acetylcholine (in the presence of L-NAME) were larger in carotid arteries from Lepr db/db mice than from DWM mice and were inhibited by indomethacin and SC560, demonstrating involvement of cyclooxygenase-1. The release of 6-ketoprostaglandin F 1α was lower in DWM mice arteries, implying lower cyclooxygenase-1 activity. Apocynin, manganese(III) tetrakis(1-methyl-4-pyridyl) porphyrin, catalase, and diethyldithiocarbamate inhibited endothelium-dependent contractions. The mRNA and protein levels of NADPH oxidase isoforms NOX1 and NOX4 were downregulated in DWM mice arteries. The in vivo and in vitro administration of lipopolysaccharide caused endothelial dysfunction in the arteries of wild-type, but not toll-like receptor 4–mutated mice. Conclusions— Toll-like receptor 4 plays a key role in obesity and diabetes-associated endothelial dysfunction by increasing oxidative stress.

The present experiments investigated whether endothelium-derived mediators modulate the effect of natriuretic peptides in porcine coronary arteries. Rings with and without endothelium were suspended in organ chambers for isometric tension recording. Concentration-relaxation curves to C-type natriuretic peptide (CNP) and atrial natriuretic peptide (ANP) were obtained during contractions to endothelin-1. Removal of the endothelium potentiated relaxations to both CNP and ANP. N(omega)-nitro-L-arginine methyl ester potentiated relaxations to natriuretic peptides only in arteries with endothelium. Sodium nitroprusside (SNP) inhibited the response to the natriuretic peptides only in the absence of the endothelium. In rings with endothelium, 1H-[1,2,4]oxadiazolo [4,3-a]quinoxalin-1-one (ODQ) and 4H-8-bromo-1,2,4-oxadiazolo[3,4-d]benz[b][1,4]oxazin-1-one (NS2028) potentiated CNP-mediated relaxations. Iberiotoxin (IBTX) reduced the response only in rings without endothelium. Glybenclamide inhibited the relaxations in both the presence and absence of endothelium. CNP-induced relaxations were reduced by 8-bromoguanosine 3',5'-cGMP (8-bromo-cGMP) to the same extent in rings with and without endothelium. There was no significant difference between the increased cGMP content caused by CNP in porcine coronary arteries with or without endothelium. In patch-clamp studies in porcine coronary arterial smooth muscle cells, the natriuretic peptide-mediated enhancement of the IBTX-sensitive big conductance calcium-activated potassium channel (BK(Ca)) amplitude was reversed by SNP and 8-bromo-cGMP. These findings demonstrate that, in the porcine coronary artery, the opening of BK(Ca) and ATP-dependent potassium channels of the vascular smooth muscle contributes to CNP-mediated relaxations. Endothelium-derived and exogenous NO inhibit the direct relaxing effect of natriuretic peptides by desensitizing the response of the BK(Ca)s of the vascular smooth muscle to the generation of cGMP.