活血丹嫩茎无性系建立的研究

为满足人们栽培活血丹对种苗的需求,以活血丹嫩茎为材料,进行了愈伤组织诱导、分化,试管苗的生根、扦插和移植的研究,成功地建立起活血丹嫩茎的无性系。结果证明:MS+BA1mg·L-1+CH10mg·L-1+2,4-D2.0-2.5mg·L-1是嫩茎愈伤组织诱导培养的理想培养基;MS+BA0.8mg·L-1+NAA0.1mg·L-1+AgN030.2mg·L-1是嫩茎愈伤组织分化培养的理想培养基;1/2MS+BA0.8mg·L-1+NAA0.1mg·L-1+GA30.4mg·L-1是不定芽继代分化培养的理想培养基;MS+IBA0.6mg·L-1+NAA0.1mg·L-1是生根培养的理想培养基。通过不定芽继代分化培养年繁殖量可达4.19.1个。移植到山林下的试管苗保持了野生活血丹的各种生物性状,且生长旺盛,当年开花结果。     

桃叶片再生不定芽的研究

以桃栽培品种曙光、金童5号和甜桃王试管苗叶片为外植体,研究了基本培养基、植物生长调节剂种类及质量浓度组合、基因型和试管苗继代次数等因素对叶片再生的影响。结果表明,适宜暗培养的培养基为LP+1.5mg·L-1TDZ+0.15mg·L-1NAA;光培养基为LP+0.5mg·L-1TDZ+0.3mg·L-1KT+0.3mg·L-1NAA和LP+0.6mg·L-1TDZ+0.3mg·L-1KT+0.4mg·L-12,4-D;金童5号和曙光具有较强的再生能力,再生率分别为21.8%和14.5%;曙光和金童5号试管苗继代第1～11次叶片的再生能力与继代前3次愈伤组织相比形成率较高;曙光再生苗在培养基1/2MS+0.5mg·L-1NAA上生根率为100%,平均生根条数为7.9。     

猕猴桃实生苗组织培养体系建立的研究

以饱满成熟的猕猴桃种子为起始材料,用 2. 5g·L-1赤霉素处理 5小时,再用次氯酸钠灭菌消毒后,将萌芽的种子接种到MS+蔗糖 20g·L-1 +肌醇 100mg·L-1和 0. 75%琼脂的培养基上,根、茎、叶生长表现良好,初步建立了猕猴桃实生苗组织培养体系。猕猴桃实生苗组织培养同利用茎段、茎尖组织培养一样,培养基中无须附加任何激素。     

白蔹的组织培养及无性系建立的研究

本文研究以白蔹为材料,进行了腋芽的生长、生长芽的分化和生根、试管苗移栽和扦插及移植于山坡栽培所需要的条件,建立白蔹腋芽的无性系。结果证明:1/2MS+IAA0.4mg·L-1是腋芽生长培养的理想培养基;MS+GA30.5mg.L-1+BA0.8mg·L-1+NAA0.1mg·L-1是生长芽分化培养的理想培养基;1/2MS+IAA0.2mg·L-1是试管苗生根培养的理想培养基;河沙与炉灰渣是白蔹试管苗移栽和扦插的理想基质;移植的试管苗具有生长旺盛、须根增加1倍左右、秋末落叶晚5-7d的特点。     

黑穗醋栗组织培养和遗传转化

通过对不同激素质量浓度配比实验,建立黑穗醋栗茎尖高频再生体系。结果表明,MS+6-BA1.0mg·L-1+NAA0.1mg·L-1培养基可高效诱导黑穗醋栗茎尖的分化,1/2MS培养基可快速诱导根的生成,形成再生植株。25mg·L-1卡那霉素可以抑制茎尖的分化,20mg·L-1卡那霉素可以抑制分化小苗的生根。利用农杆菌介导法将osMAPK4基因转化黑穗醋栗,获得了抗卡那霉素的再生植株。提取转基因植株基因组DNA,进行PCR检测,结果表明osMAPK4基因已经被整合到黑穗醋栗基因组中。     

满天星生根培养的研究

以满天星小花型品种‘仙女’作为试验材料 ,在MS培养基上加不同浓度的PP333和NAA组合 ,将不定芽平放或直立于培养基上进行培养 ,研究其生根情况。结果表明 :在所有NAA和PP333组合的培养基上 ,平放的不定芽生根速度、生根量和生根率均高于直立的不定芽 ;外源激素浓度以NAA 0 .0 1mg·L- 1 PP3330 .10mg·L- 1为最佳。     

三倍体西瓜组培快繁研究

本文探索三倍体无籽西瓜种子消毒工作程序并培养无菌苗 ,采用顶芽诱导腋芽丛生和近轴端子叶诱导以器官型再生方式产生不定芽两种途径增殖 ,诱导培养基配方为改良MS +6 -BA2 (mg·L-1,下同 ) +NAA 0～0 1,筛选出增殖培养基配方为改良MS +6 -BA1～ 2 +IBA 0 2 ,生根培养基配方为改良MS +IBA 0 3～ 0 5。采用添加KT2～ 5和GA3 1～ 2培养导致瓶内开花现象     

适宜北美花楸茎段不同组织培养阶段的培养基研究

本试验以MS作为基本培养基,添加不同种类、不同浓度的生长调节剂,寻求适宜于北美花楸茎段各组织培养阶段的培养基。研究结果表明:在初代培养阶段,其适宜的诱导培养基为MS+BA1.5+NAA1.0;继代增殖阶段,以MS+BA1.0+NAA0.5为培养基效果较好;生根阶段,最适宜的培养基为1/2MS+NAA0.1+活性炭4g/l。     

东北百里香组培再生体系的建立

 以中国特有地被植物东北百里香为试材,研究了植物生长调节剂组合对腋芽萌发和茎段、叶片外植体愈伤诱导和不定芽分化的影响。结果表明：百里香茎段腋芽可直接诱导萌发,在MS + 6-BA 0.5 mg · L-1 + NAA 0.1 mg · L-1培养基上萌发率最高,达74%,在MS + 6-BA 0.5 mg · L-1 + NAA 0.1 mg · L-1的培养基上增殖倍数为36.43。由茎段萌发的组培苗在1/2MS + IBA 0.5 mg · L-1的培养基中20 d后生根率100%,移栽成活率76.7%。继代培养中叶片外植体可以诱导出愈伤组织,但是不能进一步分化成苗。茎段愈伤诱导的最适培养基为MS + 6-BA 0.5 mg · L-1 + 2,4-D 1.0 mg · L-1,再分化培养基为MS + 6-BA（0.1 ~ 1.0）mg · L-1 + GA3（0.1 ~ 0.5）mg · L-1,分化率为33.3%。     

柿离体繁殖研究

以成年柿树休眠芽为外植体,研究了影响柿树离体繁殖的有关因素。结果表明：附加ZT0．5～2mg·1－1和IAA0．05～0．1mg·1-1或仅加ZT2mg·1-1的改良MS培养基对柿芽的增殖和生长最为适宜。在继代培养中,适当缩短每代培养时间可加快新梢的增殖速度。IAA诱导生根的效果优于IBA,用生长素溶液浸新梢基部后转入1／2MS中对生根更有利。在较低温度下,以腐殖土作基质,试管苗的移栽成活率可达88．6％。     

Effect of Sini decoction on GST expression in the ischemic myocardium

AIM:To detect the effect of Sini decoction on glutathione S-transferase (GST) mRNA expression in the ischemic myocardium. METHODS:Kunming mice were randomly divided into control group, ischemic group and Sini decoction group. Total RNA was extracted from the myocardium of mice in each group. The effect of Sini decoction on the expression of GST gene was detected by RT-PCR. RESULTS:The expression of GST mRNA in Sini decoction group was significantly up-regulated compared with the ischemic group and control group.CONCLUSION:Sini decoction can promote the expression of GST gene,which may be related to its protective effect on ischemic myocardium.     

Cortical gene expression pattern in rat focal cerebral ischemia

AIM: To investigate the genes differential expression in cortex during rat focal cerebral ischemia.METHODS: cDNA microarray chips containing numerous cDNAs were used to investigate the gene expression pattern between samples of focal cerebral ischemia and sham-control operation rats. RESULTS: Two hundred and eleven genes differentially expressed were screened out, among these genes, up-and down-regulated genes were 199 and 12, respectively. CONCLUSIONS: The analysis of gene expression pattern of focal cerebral ischemia based on cDNA microarray can realize high-throughput screening of the genes associated with the focal cerebral ischemia. The differential expression of genes may be related to the pathogenesis of focal cerebral ischemic diseases.     

Pretreatment with Sini decoction induces delayed preconditioning in rat heart and role of p38 MAP kinase

AIM: The present study was designed to determine whether Sini decoction (SND), a traditional Chinese medicine, induces delayed preconditioning-like effect in rat heart and the possible mechanism by which ischemia myocardium is protected. METHODS: Sprage－Dawleyt rats underwent three 5 min episodes of preconditioning ischemia 24 h prior to the global ischemia and reperfusion in ischemic preconditioning/second window of protection (IPC/SWOP) group or were pretreated with Sini decoction (5 mL·kg-1·d-1 for 3 days, the last treatment 24 h before global ischemia and reperfusion) in SND group. Myocardial infarct size, CK, LDH and NO were examined. p38 MAPK and PKC were determined by immunohistochemisty. RESULTS: Myocardial infarct size was significantly decreased, CK and LDH were decreased in the serum, NO2-/NO3- was increased in myocardial tissue in SND group as well as in IPC/SWOP group (there was no difference between the two groups). The expression of p38 MAPK and PKC were upregulated in myocardial tissue in SND and IPC/SWOP groups. CONCLUSION: These results suggest that Sini decoction induces delayed preconditioning-like effect in the rat heart, possibly via inducing p38 MAPK activation.     

《蔬菜学》

AIM: To investigate the mechanisms of the anti-myocardial ischemia of Sini decoction at the level of proteome. METHODS: The total proteins of left ventricular muscle were separated with two-dimensional electrophoresis. The 2-DE images were analyzed with the PDQuest7.1.1 software package. The peptide mass fingerprinting(PMF) was made with MALDI-TOF-MS, then the identification and function of the protein was made via bioinformatics. RESULTS: Sini decoction could regulate the expression of 13 proteins in the ischemic myocardium. The proteins could allocated into five groups: energy metabolism-associated proteins, signal-associated proteins ,self-repair-associated proteins, anti-free radical injury-associated proteins and myocardial contractile-associated proteins. CONCLUSION: Sini decoction could prevent the myocardium from ischemia via regulating expression of different proteins.     

Influence of ovariectomy and estrogen replacement on gene expression profile of myocardium in rats

AIM:To investigate the influence of ovariectomy and estrogen replacement treatment on profile of gene expression in myocardium by cDNA microarray,and to characterize the targeting genes of estrogen.METHODS:cDNA microarray containing 1 400 rat cDNAs was used to study the genes differentially expressed in myocardium between sham (Ⅰ),ovariectomy (Ⅱ,OVX) and estrogen replacement treatment (Ⅲ,OVX+E2) group.Then down-regulated genes in myocardium of OVX rats were further confirmed by RT-PCR.RESULTS:177 genes were differentially expressed in myocardium between sham and OVX rats,with 91 genes up-regulated and 86 genes down-regulated in OVX rats.164 genes were differentially expressed in myocardium between OVX and OVX+E2 rats,with 113 genes up-regulated and 54 genes down-regulated in OVX rats.There were 54 genes differentially expressed in OVX compared to sham and OVX+E2.They are involved in membrane channels and transporters (18),cell receptors (9),intracellular transducers/effectors/modulator (7) and metabolism (6).Most of the genes (45) were down-regulated in OVX rats and up-regulated in OVX+E2 rats.RT-PCR test confirmed the results of cDNA microarray.CONCLUSIONS:Long-term estrogen replacement may influence the expression of genes involved in membrane channels and transporters,cell receptors,intracellular transducers/effectors/ modulator and metabolism.Long-term estrogen replacement has some beneficial effects on ionic concentration and cardiac function which partially comes from the results of influence of expression on Na+,K+-ATPase and Na+/H+ exchanger.Estrogen has an inhibitory effect on the expression of dopamine receptor,which partially clarify the myocardial protection of estrogen.     

Effects of lovastatin on differentially expressed genes in HepG2 cells

AIM: To analyze the lovastatin-induced differential gene expression in HepG2 cells using a cDNA microarray assay. METHODS: Total RNA was extracted from the lovastatin-treated HepG2 cells and control group. cDNA was synthesized from RNA with Cy3/Cy5-labelled dCTP. Then the hybridization was conducted. The result was analyzed using Imagene and Genespring software. RT-PCR was carried to confirm the hybridization results. RESULTS: 30 genes were up-regulated while 11 genes were down-regulated in lovastatin-treated HepG2 cells, involved in some major functional areas including signal transduction, cell cycle regulation, tumor immunity, and so on. CONCLUSION: The analysis of differentially expressed genes in lovastatin-treated HepG2 cells is helpful to explore the mechanism of the anti-tumor activity of statins.     

Effect of Sini decoction on expression of caveolin-1 and eNOS in EAhy926 cells injured by homocysteine

AIM：To investigate the mechanism of Sini decoction in treating human vascular endothelial cell injury and the roles of caveolin-1 and nitric oxide (NO) system in this procedure. METHODS：Model of human umbilical vein endothelial EAhy926 cells injured by homocysteine (Hcy) was established. The protective effect of Sini decoction on the injured EAhy926 cells was observed, and the expression of caveolin-1 and endothelial nitric oxide synthase (eNOS) was detected by real-time fluorescence quantitative PCR and Western blotting. RESULTS：Compared with control group, the Hcy-treated EAhy926 cells showed reduced adherent cell number and NO concentration in culture supernatant, decreased expression of eNOS mRNA and protein, and increased expression of caveolin-1 mRNA and protein (all P<0.05). Compared with Hcy group, better growth of adherent cells, elevated NO concentration in culture supernatant, attenuated expression of caveolin-1 mRNA and protein, and enhanced expression of eNOS mRNA and protein in Sini decoction groups were observed (all P<0.05). CONCLUSION：Homocysteine may injure EAhy926 cells by enhancing the expression of caveolin-1 and suppressing the expression of eNOS, while Sini decoction may protect EAhy926 cells by suppressing the expression of caveolin-1 and enhancing the expression of eNOS.