Experimental studies on anti-atherosclerosis effects of Momordica charantia L in rabbits

AIM: Anti-atherosclerosis effects of Momordica charantia L was further studied in a New Zealand rabbit atherosclerotic model at the basis of anti-inflammation and antioxidant effects. METHODS: Animals were divided into 3 groups: normal group (normal rabbit diet), atherosclerosis group(diet containing 2% cholesterol), and Momordica charantia L group(diet containing 2% cholesterol and 1.5% sarcocarp of Momordica charantia L ). Ninety days later, all animals were sacrificed. The effect of Momordica charantia L on atherosclerosis was evaluated by measuring serum lipid and total cholesterol content of artery wall, observing fatty liver degree, aorta arteriosclerotic area, and the thickness of intima. RESULTS: The level of total serum cholesterol and LDL-C in Momordica charantia L treatment group were obviously lower than those in atherosclerosis group, so were the total cholesterol content of artery wall, fatty liver degree, atherosclerotic area, intima thickness and I/M ratio, but no significantly difference was found between the two groups in TG level. The level of HDL-C in Momordica charantia L treatment group was evidently lower than that in normal control group. CONCLUSION: Momordica charantia L has an anti-atherosclerosis action in rabbits.     

Role and mechanism of endothelial hydrogen sulfide in anti-atherosclerosis effect of estrogen

AIM：To investigate possible role and mechanism of endothelial hydrogen sulfide (H2S) in the anti-atherosclerosis effect of estrogen. METHODS：For in vitro experiment, cultured human umbilical vein endothelial cells (HUVECs) were treated with 17β-estradiol (E2), estrogen receptor α (ERα) agonist propylpyrazole triol (PPT), estrogen receptor β (ERβ) agonist diarylpropionitrile (DPN) and estrogen receptor antagonist ICI 182780 (ICI) for 5 min, and then the concentration of H2S in cell culture supernatants was detected by sensitive sulphur electrode assay. For in vivo experiment, high-fat diet-fed and ovariectomized (OVX) female ApoE–/– C57BL/6 mice were randomly divided into 3 groups and treated with placebo (OVX group), E2 (OVX+E2 group) and E2 plus cystathionine γ-lyase (CSE) inhibitor DL-propargylglycine (PPG) (OVX+E2+PPG group), respectively, and the blood concentration of H2S and the atherosclerotic plaque size were detected 8 weeks later. RESULTS：E2 significantly enhanced the release of H2S from HUVECs in a concentration-and time-dependant manner which could be blocked by the administration of ICI. Meanwhile, ERα agonist PPT, not ERβ agonist DPN, demonstrated similar effects to E2. Compared with OVX group, the atherosclerosis in mice was attenuated and the blood concentration of H2S was elevated in OVX+E2 group, while no significant changes were detected in OVX+E2+PPG group. CONCLUSION: Endothelial H2S can be rapidly released when membrane ERα is activated by estrogen, which plays a very important role in the anti-atherosclerosis effect of estrogen.     

丘陵地成龄低产山楂园改造技术

1997-2000年对12年生山楂低产园进行技术性质改造，重点抓好土壤深翻改良，增施肥料，精细修剪，病虫害防治等措施，使666.7m^2产量由1996年改造前的640kg增加到2000年的1986kg，效益显著提高。     

山楂酱的简易制作

山植酱酸甜可口，营养丰富，色释诱人，是佐餐佳品。且山楂来源广，价格较低。适合家庭制作，现将生产过程介绍如下：     

成龄山楂密植园低产改造技术

自2002年开始,对1985年定植、株行距2m×3m的低产山楂园进行改造,改造后产量逐年上升。2003年666.7m2产量1254kg,2004年2212kg,2005年达2635kg,比改造前(2002年)增产3倍多,经济收入增加6倍以上。主要技术措施:深翻扩穴,浅刨树盘,增施有机肥,及时追肥浇水,全园覆草,合理间伐,疏大枝,回缩冗长枝,冬剪结合培养紧凑枝组充实内膛,落头开心控制树高,刮树皮,疏花序,花期喷施山楂丰收素提高坐果率和及时防治病虫害等。     

山楂幼树的增枝促花技术

1　增加前期枝量的技术为增加山楂幼树的前期枝量 ,除轻剪缓放外 ,还可用拉枝、拉干、开张角度、摘心、刻芽、环割等促进增枝。拉枝开角 :此法用于直立旺长枝、角度小的骨干枝和辅养枝。这类枝生长快、发枝少、后部空。可在发芽前用绳将 1ｍ以上的枝条拉向空间较大的地方 ,或对一、二年生强旺枝从基部向先端捋、拿 1～ 2次。6月上中旬至 7月上中旬拉枝开角最适宜。具体做法是 :将 1ｍ以上的长枝慢慢弯曲 ,使梢端接触地面 ,用土压实。土压部位之前留 4～ 6片叶。压枝一个月后 ,待枝条角度基本固定 ,再从土中扒出。摘心 :主要对象是生长旺的当…     

山楂过氧化物酶基因的克隆及在烟草中异位表达分析

【目的】从山楂(Crataegus pinnatifida)中克隆过氧化物酶(POD)编码基因,通过转基因验证其在木质素合成中的作用,为揭示山楂内果皮形成分子机制奠定基础。【方法】利用RT-PCR技术从山楂克隆基因,以p RI 101-AN为构建基因过量表达载体的基础载体,通过农杆菌介导的遗传转化方法将目的基因导入烟草。【结果】从山楂中克隆出编码序列长度为996 bp的POD72基因,命名为Cp POD72。山楂POD72与木本和草本植物POD72的氨基酸序列一致性均较高。构建了Cp POD72基因的过量表达载体,通过农杆菌介导的转化获得了66个烟草转化植株。RT-PCR检测结果显示Cp POD72基因在烟草转基因植物中异位表达,组织化学染色分析表明转Cp POD72基因烟草植株中木质素含量增加。【结论】Cp POD72基因可能参与木质素合成。     

雾灵红山楂的主要性状及栽培技术要点

雾灵红为河北省兴隆县林业局于1988年选育并通过省级鉴定的山楂优良品种。该品种果大质优，丰产，盛果期666．7m^2产量达1500kg以上，适应性、抗寒性和抗病性强。最适宜在光照充足、排水良好、土层深厚肥沃的坡地栽培。建园时配好授粉树、盛花期及时疏花序可明显提高花朵坐果率，果大而整齐。整形期间多留辅养枝可显著增产。     

潍坊市的板栗、柿和山楂生产现状

阐述了目前山东省潍坊市的板栗、柿和山楂分布的情况和生产规模,以及在生产中存在的主要问题,提出了"十一五"期间的发展规划,并就此对潍坊市板栗、柿和山楂示范园与生产基地的建设提出几点建议.     

山楂白粉病的发生和防治技术

山楂白粉病以子囊壳在落叶上越冬，在山楂展叶期开始侵染危害，新梢生长，开花，坐果期是病害流行期，新梢停止生长，高温来临时，子囊壳开始形成，侵染危害渐减轻，防治山楂白粉病，在秋冬彻底清除落叶，杜绝病原的基础上，于新梢生长期喷布2－3次粉锈宁等即可控制危害。     

响应面优化虎杖固态发酵工艺条件的研究

应用产β-葡萄糖苷酶的黑曲霉固态发酵酶解虎杖中的虎杖苷,以提高虎杖中白藜芦醇的含量。采用均匀实验设计和响应面分析方法对影响虎杖中白藜芦醇含量的固态堆积发酵工艺条件(温度、湿度、菌龄和接种量)进行优化。结果表明:最佳的固态发酵工艺条件为:温度42℃,湿度1.85∶1、菌龄42h、接种量1.8%,在此条件下,发酵48h后,白藜芦醇含量提高为1.52%,是未进行固态发酵虎杖中白藜芦醇含量的4.47倍。优化固态发酵工艺条件能较大程度的提高从虎杖中获取白藜芦醇的得率。     

山楂属植物中苹果褪绿叶斑病毒的RT-PCR检测及病毒脱除方法

【目的】建立优化的山楂属植物苹果褪绿叶斑病毒(ACLSV)的RT-PCR检测体系,获得以山楂组培苗为试材的高效ACLSV脱除方法。【方法】基于转录组高通量测序解析的山楂属植物ACLSV基因组序列设计病毒检测引物,比较c DNA模板浓度、Taq DNA聚合酶种类及采样部位对病毒RT-PCR检测效果的影响。通过组培苗热处理和培养基中添加三氮唑核苷来脱除山楂植株中的ACLSV。【结果】基于转录组高通量测序获得的2个ACLSV山楂分离物的全基因组序列设计并筛选出适合于山楂属植物ACLSV的RT-PCR检测引物。建立了优化的山楂属植物ACLSV的RT-PCR检测体系。对80份山楂样品进行病毒检测,其中有18个样本检测出ACLSV,带毒率为22.5%。比较了热处理(37℃、30d)、化学处理(培养基中添加20 mg·L-1的三氮唑核苷,处理40 d)、热处理与化学处理结合方法(处理30 d)3种方法对山楂组培苗中ACLSV的脱除效率,表明热处理或热处理与化学处理结合的方法能够完全脱除山楂中的ACLSV,而单独化学处理的脱毒率不能达到100%。【结论】建立了优化的山楂属植物ACLSV的RT-PCR检测体系,表明组培苗热处理是脱除山楂植株中ACLSV的有效方法。     

山楂果实发育期矿质元素和色素含量的动态变化及相关关系

以大黄面楂、上口、左伏3号、星石榴、甜水5个山楂品种为试材,研究山楂果皮色泽形成与自身矿质元素含量的关系,结果表明,在果实发育过程中,叶绿素含量先上升后下降,类胡萝卜素含量持续下降,花青素含量除大黄面楂外,在果实着色期迅速上升。上口、左伏3号、大黄面楂N含量与果皮花青素含量相关性不显著,甜水和星石榴呈乘幂函数曲线极显著负相关;左伏3号和大黄面楂Ca含量与果皮花青素含量相关性不显著,甜水、星石榴、上口3个品种呈乘幂函数曲线显著或极显著负相关;左伏3号、大黄面楂Fe含量与果皮花青素含量相关性不显著,甜水、星石榴、上口呈乘幂函数曲线显著或极显著负相关;各个品种K元素含量和果皮花青素含量相关性均不显著。     

养根控冠促花技术对山楂树产量的影响试验

莱芜市地处鲁中地区,而我市的山楂园多种植在土地条件差的山丘薄地,水源奇缺,保肥水能力低。针对这一情况,我们在2004-2006年对6-8年生山楂树进行了小区试验,试验面积为667m2,株行距为3m×4m,分别进行了深翻扩穴、改良土壤,增肥养根,整形修剪,激素控冠等养根控冠促花技术,试验前的2003年小区平均产量667m2为976kg,试验期间的2004—2006三年平均667m2产2687kg,试验最后一年2006年平均667m2产3000kg,好果率达96%,达到丰产丰收,现将综合管理技术总结如下。1、深翻扩穴、改良土壤全园深翻扩穴,深度50-80cm,底层压20-30cm的杂草、麦秸等,上覆一层…     

PPARγ gene and protein expression in aortic plaque of different week-old apoE-knock out mice and the relationship with the composition of aortic plaque

AIM: To investigate the relationship of PPARγ gene expression with the composition of aortic plaque in apoE-knock out mice. METHODS: PPARγ gene and protein in aortic area of 20-week-old and 40-week-old apoE-knock out mice were investigated using RT-PCR and immunoblotting. The same aged wild type mice (C57BL/6J) were served as control (n=10). The composition of aortic plaques was analyzed by Movat method and oil red O staining. The expression of antigens such as PPARγ, SM-actin and MOMA-2 in aortic plaque were compared using immunohistochemistry. The relationship of PPARγ with macrophage, smooth muscle cells (SMC), lipid, elastic fiber, collagen and proteoglycan in aortic plaque were analyzed using immunofluorescence. RESULTS: PPARγ gene and protein in aortic wall and plaque of apoE-knock out mice were more significant than that in the same aged C57BL/6J mice (P<0.05). PPARγ expression at 40-week-old apoE-knock out mice was most significant and very low in C57BL/6J mice. More PPARγ expression of gene and protein at 20-week-old C57BL/6J mice than 40-week-old C57BL/6J mice were observed. Compared with 20-week-old apoE^-/- mice, the lipid pool in aortic plaque at 40-week-old apoE^-/- mice were increased remarkably, while elastic fiber, collagen and proteoglycan in plaque were decreased and aortic remodeling was very significant. Even, upregulation of MOMA-2 and downregulation of SM-actin were also detected in latter (P<0.05). In addition to SMC of aortic tunica media, PPARγ also expressed in SMC and macrophages in the aortic plaque of apoE^-/- mice. PPARγ was very enriched in lipid pool of the plaque. CONCLUSION: PPARγ expression level decreases with aging in C57BL/6J mice, while increases with plaque progression in apoE-knock out mice. There is positive correlation between PPARγ expression and lipid composition in plaque. The observed upregulation of PPARγ gene expression in aortic plaque may be a compensatory behavior and protective mechanism in apoE-knock out mice.     

Effects of herbs of activation blood on atherosclerotic plaque morphology in ApoE gene-deficient mice

AIM: To observe the effects of six common traditional Chinese herbs of activating blood, paeoniae rubra radix, salviae miltiorrhizae radix, ligustici, rhizome, notoginseng radix, pruni persicae semen and wine staemed radix et rhizome, on atherosclerotic plaque structure and stabilization in ApoE gene-deficient mice. METHODS: Four sections of the aortic root were choosen and stained with hematoxylin and masson. All sections were measured with Image-Pro Plus Version 4.5.1 (IPP) software. RESULTS: Compared with the model group, plaque area corrected by cross-sectional vessel wall area reduced significantly in salviae miltirrhizae radix treatment group, lipid core area reduced in paeoniae rubra radix group, pruni persicae semen and wine steamed radix et rhizome treatment group, minimum thickness of fibrous cap became thicker significantly in salviae miltiorrhizae radix, ligustici, rhizome, pruni persicae semen and wine steamed radix et rhizome treatment group. CONCLUSION: These Chinese herbs may stabilize the atherosclerotic plaques in ApoE gene-deficient mice by interfering their structure, but their effects do not parallel with their activating blood efficacy in traditional Chinese medicine.     

Experimental study on induction of atherosclerotic plaque instability in rabbits after transfer of wild-type p53 gene

AIM: To study the apoptotic role of wild-type p53 in induction of plaque instability in atherosclerotic rabbits. METHODS: Fifty-four New Zealand White rabbits underwent balloon-induced abdominal aortic wall injury and then were fed on a diet of 1% cholesterol. At the end of the eighth week, the rabbits were randomly divided into two groups: group A and group B. Recombinant adenovirus carrying p53 and β-galactosidase (LacZ) genes were injected in group A and B, respectively. Two weeks later, 10 rabbits each in group A and B was killed and the remaining rabbits all underwent pharmacological triggering with injection of Chinese Russell's viper venom and histamine. RESULTS: Compared with group B, p53 gene over-expression in group A resulted in a marked increase in number of positive apoptotic cells (2.5%±0.8% vs 1.0%±0.3%, P<0.05) and a significant decrease in vascular smooth muscle cells (47.5%±6.8% vs 80.4%±10.6%, P<0.01), the thickness of the fibrous cap ［(132.9±56.7）μm vs （181.8±59.7） μm, P<0.05］ and the cap/intima-media ratio (0.20±0.18 vs 0.21±0.11, P<0.05). CONCLUSION: Transfer of human wild-type p53 genes effectively promotes apoptosis of VSMCs in atherosclerotic plaques, which makes the plaques vulnerable to rupture.     

Effects of Shuxuetong combined with atorvastatin on LDL-C,TG, IMT and plaque area in transient ischemic attack patients

AIM:To observe the therapeutic effect of Shuxuetong combined with atorvastatin for treating transient ischemic attack(TIA). METHODS:We retrospect 46 cases of TIA patients from August 2009 to December 2010 in our hospital treated with Shuxuetong combined with atorvastatin. The clinical data such as coagulation function, blood lipid, fasting blood glucose and the carotid color Doppler ultrasonic examination before and after treatment were collected. All patients continued to take atorvastatin for 6 months and were followed up for 6 to 12 months. RESULTS:The total effectiveness of using Shuxuetong combined with atorvastatin for TIA in 46 cases was 91.30%. Five patients were found to suffer from recurrent TIA or the occurrence of stroke in the cured or effective 42 cases during follow-up, the incidence of which was 11.90%. Six months after treatment, low-density lipoprotein-cholesterol(LDL-C) and triglycerides(TG) were obviously lower than those before treatment(P<0.05), and the carotid intimal thickness(IMT) and the plaque area were decreased as compared with those before treatment(P<0.05). CONCLUSION:Treatment with Shuxuetong+atorvastatin for TIA has good therapeutic effect and reduces the recurrence of TIA and stroke incidence, which may be related to the reduction of LDL-C and TG levels, IMT and the plaques area.     

Changes of von Willebrand factor and vascular-cellular adhesion molecule-1 in patients with unstable angina

AIM: To explore the changes of von Willebrand factor (vWF) and soluble vascular-cellular adhesion molecule-1 (sVCAM-1)， and the correlation of vWF, sVCAM-1 with acute myocardial ischemia. METHODS: By means of enzyme linked immunosorbent assay (ELISA)， the changes of vWF and sVCAM-1 were measured in 73 patients with unstable angina (UA) during angina attack and relief period (24 h after attack) as well as in 50 controls. RESULTS: The levels of vWF and sVCAM-1 were significantly higher in UA patients than those in controls (P<0.01). The increase in vWF and sVCAM-1 in angina attack was more obvious than those in relief period (P<0.01). vWF correlated positively with sVCAM-1 (r=0.785， r=0.674，P<0.01). In different types of angina, there were significant difference of vWF，sVCAM-1 between angina attack and relief periods (P<0.01)， and increases in all the indicators in spontaneous angina patients were more obvious than those in exertional and postinfarction angina patients. CONCLUSION: Acute myocardial ischemia might be related to the changes of vWF and sVCAM-1.