应用生物反应器扩繁‘Casa Blanca’百合鳞茎

 在生物反应器内‘Casa Blanca’百合鳞茎的生长显著优于在固体培养基培养，培养16周后鳞茎鲜样质量为培养初期的22．4倍，但固体培养只达到16．5倍；5 L气球型生物反应器内接入400个小鳞茎时，可获得295个>1．1 g以上的鳞茎，多于其它培养密度；生物反应器内生产的鳞茎地上茎发生率高于在体培养基上培养的鳞茎，1．1 g以上的达到90％，但在固体培养基培养的鳞茎，2．1 g以上的也只达到65％。     

蝴蝶兰离体培养条件的筛选

应用正交设计法研究了基本培养基、生长物质和蔗糖对蝴蝶兰花梗腋芽丛生芽诱导、增殖和生根培养的影响。筛选出最佳基本培养基为1／2MS，最佳诱导培养基为“1／2MS BA5．0mg／L NAA0．5mg／L”，最佳增殖培养基为“1／2MS BA3．0mg／L NAA0．5mg／L十蔗糖30g／L”，最佳生根培养基为“1／2MS NAA0．2mg／L IBA0．1mg，L 蔗糖20g／L”。     

金耳的液体发酵研究

研究表明，金耳(Tremella aurantialba)液体发酵最适培养基组成为玉米粉2％，蔗糖3％，酵母膏0．1％，KH2PO40．1％，MgSO40．05％；培养基最佳初始pH、250mL三角瓶装量、接种量、种子菌龄分别是3．5、80mL、10％和120h；用10L发酵罐发酵时，最佳培养条件是种子种龄144h、28℃、接种量10％、通风量2．5L／min。     

河阴‘软籽石榴’组培快繁技术研究

以河阴‘软籽石榴’的当年生嫩枝茎段为外植体进行组织快繁试验。结果表明：河阴‘软籽石榴’的最佳诱导培养基为MS＋NAA0．3mg，L＋BA0．5mg／L，增殖培养基为MS＋BAl．0mg／L＋NAA0．2mg／L，生根培养基为1／2MS＋IBAl．5mg／L＋NAA0．2mg／L，生根率达到96％。     

利用间歇浸没式生物反应器进行香蕉(Musa AAA Cavendish var.Williams B6)组培快繁研究(英文)

以香蕉(MusaAAACavendishvar.WilliamsB6)茎尖诱导的组培苗为材料,开展了利用间歇浸没式生物反应器(TIBS)进行组培快繁技术体系的研究。结果表明,(1)TIBS系统可使香蕉组织培养一代增殖14倍以上,比传统方法提高2～3倍;(2)含2.0mg·L-16-BA的TIBS系统适合于增殖培养;(3)TIBS系统中,以第7代继代材料为宜,接种密度在每株100mL培养基最有利于香蕉组培苗的增殖和生长;(4)TIBS系统中浸没间歇频率在4min/3h时增殖和生长表现比较优异,而4min/6h时则有利于根的诱导和根的生长。     

果梅胚离体培养研究

本试验以优选果梅‘黔荔1号’单株成熟胚为材料，对其进行离体培养，并对外植体灭菌时问、胚萌发和增殖以及生根培养进行研究。结果表明：以0．1％HgCl2消毒8min效果最好，其存活率为75．0％；适合胚萌发和增殖的基本培养基为1／2MS＋2．0mg／L6-BA＋1．0mg／LNAA＋3．0mg／LGA3和1／2MS＋2．0mg／L6-BA＋0．5mg／LNAA＋1．0mg／LGA3；适合生根的培养基是1／2MS＋0．5mg／LIBA，生根率为91％，平均根数达到3．1。     

甜樱桃与中国樱桃杂种的胚抢救及杂种鉴定

 以甜樱桃为母本，中国樱桃为父本，进行种间杂交，建立了杂种胚抢救技术体系，并对部分 杂种进行了鉴定。结果表明：(1)母本中，‘养老’坐果率最高，‘养老’在授粉4周后胚发生败育，而‘那翁’与‘红灯’则是在第3周后败育；(2)最佳萌发与生长培养基为1／2 MS+BA 2 mg／L+IAA 1 mg／L+维生素c 10 mg／L，最佳继代增殖培养基为1／2 MS+BA 2 mg／L+IAA 1 mg／L+NAA 0．1 mg／L+维生素c 10 mg／L，最佳生根培养基为1／2 MS+IBA 0．2 mg／L+维生素C lOmg／L；(3)S-allele specific PCR、RAPD及叶片形态鉴定结果证实了杂种的真实性.     

‘火鸟’蝴蝶兰花梗组织培养技术的研究

试验目的在于选出合适的‘火鸟’蝴蝶兰花梗组织培养的培养基配方,从而建立其组织培养技术体系。试验采用带腋茅的花梗段来诱导丛生芽,继而进行继代增殖培养,最后诱导生根。对培养的各阶段培养基及激素配比进行研究,结果表明,较好诱导培养基为MS＋6-BA5．0mg／L＋NAA0．05mg／L,继代增殖培养基为MS＋6-BA2．0mg／L＋NAA0．2mg／L,生根培养基为1／2MS＋NAA0．3mg／L。     

不同激素对春石斛的组织培养影响研究初报

用春石斛（Dend robium nobile）1～3mm的茎尖为外植体，以1／2MS为基本培养基，调节不同激素水平诱导芽体萌发，进行繁殖培养。研究结果表明：萌芽诱导阶段：1／2MS＋6-BA0．5mg／L＋NAA0．5mg／L＋椰子汁200rnL／L＋蔗糖30g／L＋活性碳3g／L＋琼脂5g／L中萌芽早，生长快；继代和增殖培养阶段：以1／2MS＋6-BA0．5mg／L＋NAA2．0mg／L＋椰子汁200mL／L＋蔗糖15g/L＋活性碳3g／L＋琼脂5g／L效果最好；生根阶段培养基：1／2MS4-NAA0．1mg／L＋蔗糖15g／L＋活性碳3g／L＋琼脂5g／L每茎段可生根4条肉质健壮，在培养过程中达到了较好的效果。     

辣椒茎尖培养脱毒研究

以云南小米椒为试材，经无菌发芽，在2～3真叶时，经35℃处理4周，取无菌苗生长点，以MS为基本培养基，在其中分别添加6-苄基腺嘌呤(6-BA)、玉米素(ZT)、细胞激动素(KT)、赤霉素(GA3)、吲哚乙酸(IAA)和萘乙酸(NAA)等生长调节剂进行培养。经筛选，发现其中MS BA7．0mg／L IAA0．05mg／L比较适宜于茎尖培养。MS ZT8．0mg／L GA32．0mg／L比较适宜于茎的伸长。将长2cm以上的芽无根苗转入1／2MS IAA0．2mg／L NAA0．1mg／L的培养基上有利于根的生长。生根后移入土壤可正常生长。1～1．5mm长的茎尖成苗脱毒率达96．3％。0．1～0．5mm长的茎尖脱毒率达100％。     

Metallothionein protects rat hepatic nuclear nucleoside triphosphatase from hydroxyl radical-induced suppression

AIM:Metallothioneins (MTs) are cysteine-rich metal-binding proteins that exert cytoprotection during metal exposure and oxidative stress. The present study was designed to investigate whether MT can directly protect NTPase on nuclear envelope from damage induced by hydroxyl radical.METHODS:Isolated hepatic nuclei from rat liver were exposed to Fe²⁺/H₂O₂ with or without MT, and the NTPase activity on nuclei was assayed using ATP and GTP as substrate, respectively.RESULTS:Incubation of rat hepatic nuclei with the Fe²⁺/H₂O₂ (in μmol·L^-1/μmol·L^-1 : 0.1/0.5, 0.5/2.5, 1/5, 5/25) resulted in a concentration-dependent decrease in nuclear NTPase activities (P＜0.01). Incubation of hepatic nuclei with different concentrations of MT (10^-9-10^-4mol·L^-1)and Fe²⁺/H₂O₂ (1 μmol·L^-1/5 μmol·L^-1) for 10 min, nuclear NTPase activities were increased in a MT concentration-dependent fashion as compared with that of incubation with Fe²⁺/H₂O₂(1 μmol·L^-1/5 μmol·L^-1) alone. When MT was at 10^-4 mol·L^-1, TNPase activities reversed to (102±10) nmol·mg^-1 protein·min^-1(for ATP as substrated) and (131±12) μmol·g^-1 protein·min^-1(for GTP as substrate), which had no significant defferences from that of the controls (112±8 and 142±10 μmol·g^-1 protein·min^-1, respectively) (P＞0.05). In addition, incubation of hepatic nuclei with only MT had no effect on nuclear NTPase activity. CONCLUSION:These data demonstrate that hydroxyl radical generated from Fe²⁺/H₂O₂ might attack nuclear NTPase. MT antagonistically reduces toxicity of Fe²⁺/H₂O₂ system to the NTPase.     

Role and regulation of calcineurin in angiotensin Ⅱ-stimulated cardiac myocyte hypertrophy of rats

AIM: To study the role and regulation of calcineurin(CaN) in angiotensin II(AngⅡ)-stimulated cardiacmyocyte hypertrophy of rats. METHODS: Using AngⅡ to induce the cultured cardiac myocyte hypertrophy of rats, and investigating the effect of CaN inhibitor on [³H]-leucine incorporation of AngⅡ-stimulated cardiomyocytes and the regulation of various factors on CaN activity in cardiomyocytes.RESULTS: AngⅡ can stimulate the CaN activity in cultured neonatal rat cardiomyocytes in a dose- and time-dependent manner. In cardiac myocytes incubated with 10, 100, 1000 nmol·L^-1 of AngⅡ for 12h, the CaN activities increased respectively by 13%,57%(P＜0.05) and 228%(P＜0.01) compared with that in non-stimulated cardiomyocytes. The CaN activities in AngⅡ-stimulated cardiomyocytes were significantly inhibited by losartan(50 μmol·L^-1), H₇(50 μmol·L^-1)and Fura-2/AM(4 μmol·L^-1),while no effect was observed with PD98059(50 μmol·L^-1).The [³H]-leucine incorporation in AngⅡ-stimulated cardiomyocytes increased by 46%(P＜0.01) compared with that in control group, which was dramatically inhibited by cyclosporin A(0.5~5μg/mL). CONCLUSIONS: Calcineurin, a Ca²⁺/calmodulin-dependent protein phosphatase, may play an important role in AngⅡ-induced cardiac myocyte hypertrophy. The activation of CaN may dependent on the sustained increases of [Ca²⁺]i and be regulated by some protein kinases (such as PKC,etc.).     

Role of leukotriene D4 in promoting proliferation of cultured human airway smooth muscle cells

AIM: To investigate whether leukotriene D₄(LTD₄) would stimulates proliferation of cultured human airway smooth muscle (ASMC). METHOD: Human ASMC were isolated and subcultured, varying concentration of LTD₄ were added to the media. Cell counts were obtained, -thymidine([³H]-TdR) incorporation and inositol 1, 4, 5-trisphosphate (IP₃) accumulation were measured. RESULTS: LTD₄(0.1nmol·L^-1~10 nmol·L^-1) increased cell number and also increased incorporation of[³H]-TdR and accumulation of IP₃ in a concentration dependent manner(P＜0.01). The latter response was blocked by phospholipase C inhibition with neomycin (1 μmoL·L^-1(P＜0.01). However, neomycin had no effect on the promitogenic action of LTD₄. CONCLUSION: LTD₄ stimulates proliferation of cultured human ASMC and may play a role in airway remodeling of asthma.     

Effects of tetrandrine and fructose-1, 6-diphosphate on the elevated intrasynaptosomal [Ca2+]i induced by excitatory amino acids

AIM: To study the effects of tetrandrine(Tet) and fructose-1, 6-diphosphate(FDP) on the elevated intrasynaptosomal [Ca²⁺]_i induced by excitatory amino acids(EAA). METHODS: A rapid method for preparing synaptosomes was used, and intrasynaptosomal free calcium([Ca²⁺]_i) was measured by using the fluorescent indicator quin-2. RESULTS: L-glutamate(Glu, 100 μmol/L), aspartate(Asp, 100 μmol·L^-1), N-methy1-D-aspartate(100 μmol/L) and Glu(50 μmol/L) plus Asp(50 μmol/L) all elevated intrasynaptosomal [Ca²⁺]_i in a dose-dependent manner. Pretreatment with Tet(10, 30, 60 μmol/L), FDP(15, 30, 75, 150 μmol/L), MK-801(10, 20 μmol/L) and Tet(15, 30 μmol/L) plus FDP(15, 30 μmol/L) all attenuated the increase in intrasynaptosomal [Ca²⁺]_i induced by EAAs mentioned as above in a dose-dependent manner, and the effect of Tet plus FDP was most significant. CONCLUSION: Both Tet and FDP inhibited a rise in intrasynaptosomal [Ca²⁺]_i induced by EAAs, which may be one of mechanisms that Tet and FDP pretect cerebral tissues against ischemia injury.     

紫玉兰花干燥方法对其活性成分和抗氧化性的影响

以紫玉兰(Magnolia liliflora Desr)花为材料,采用晒干、阴干、热风干燥、微波干燥等不同的干燥方法进行处理,探索其对紫玉兰花黄酮、多酚含量及抗氧化性的影响。结果表明:不同干燥方法处理紫玉兰花,干燥速度快慢顺序为:微波干燥热风干燥晒干阴干。阴干、900 W微波干燥和50℃热风干燥样品的黄酮、多酚含量及抗氧化性较高,晒干处理对多酚、黄酮的分解损失最大,抗氧化活性最低。DPPH·清除率与黄酮含量(R2=0.7300)和多酚含量(R2=0.6675)显著相关,总还原力与多酚含量显著相关(R2=0.8234)。     

Expression of peroxisome proliferator-activated receptor α in rat fatty liver

AIM:To investigate the role of expression of peroxisome proliferator-activated receptor α(PPAR α) in pathogenesis of rat fatty liver.METHODS:The rats were treated with a low dose of carbon terachloride (CCl₄) and fed a high fat diet to produce fatty liver. We determined the concentrations of triglyceride (TG), total cholesterol (TC), free fatty acid (FFA) in liver and the alanine aminotransferase (ALT) activity, tumor necrosis factor-α (TNF-α), FFA in serum and the degree of hepatocytic steatosis. Total RNA of liver was extracted, and the expression of PPAR α were analyzed by semi-quantitative RT-PCR method.RESULTS:In model group, the hepatocytic PPAR α mRNA expression decreased to 0.41±0.28, compared to 1.41±0.29 in the control group (P＜0.01). The contents of TG, TC, FFA in model rat liver were (1.88±0.20) mmol·L^-1, (11.03±1.12) mmol·L^-1 and (1 260.38±151.27) μmol·L^-1, respectively, compared to (0.53±0.10) mmol·L^-1, (1.25±0.25) mmol·L^-1 and (334.30±27.09) μmol·L^-1 in the control group (P＜0.01). The activity of ALT, concentrations of TNF-α and FFA in serum were also increased remarkably in model group.CONCLUSION:Oxidation of fatty acid and utilization of lipids in liver are affected by reducing the expression of PPAR α, which result lipid accumulation in liver.     

光强对黑豆芽苗菜生长和营养品质的影响

采用发光二极管（light emitting diode，LED）精确调制光强，研究不同光强对黑豆芽苗菜生
长和营养品质的影响。结果表明：与黑暗培养相比，光强为3、9、15 μmol·m^-2·s^-1 时黑豆芽苗菜的下胚
轴直径显著增加；光强为3 μmol·m^-2·s^-1 时黑豆芽苗菜的VC 含量显著增加；光强为9 μmol·m^-2·s^-1
时黑豆芽苗菜的可溶性蛋白、可溶性糖和蔗糖含量显著增加， 并且POD 活性显著提高。在3～15
μmol·m^-2·s^-1 的光强范围内，黑豆芽苗菜的叶绿素a、叶绿素b、叶绿素a+b 及类胡萝卜素含量均显著
高于黑暗培养，并且各色素含量均随着光强的增大而显著增加。总体而言，3～9 μmol·m^-2·s^-1 的光强
培养有利于黑豆芽苗菜的生长和部分营养品质的改善。     

Effects of cardiac remodeling on atrial arrhythmogenesis in aged rabbit left atrium

AIM: To investigate the effects of myocardial remodeling of aged left atrium (LA) on atrial arrhythmogenesis in rabbits.METHODS: The male New Zealand rabbits were divided into young LA and aged LA groups. By observing the changes of monophasic action potential (MAP) and burst-pacing in LA of the rabbits in vivo, the main cardioelectrophysiological parameters such as resting membrane potential (RMP), action potential amplitude (APA), ma-ximum rise veloctiy of action potential (dv/dt_max), plateau potential and action potential duration at 30%, 50% and 90% (APD₃₀, APD₅₀ and APD₉₀), as well as the inducibility and duration of atrial arrhythmias were recorded. L-type calcium current (I_Ca,L) was analyzed via whole-cell patch-clamp technique in enzymatically dissociated single rabbit LA myocytes. The myocardial collagen content was quantified after Masson staining, and the ultrastructure of the LA cells was observed under scanning electron microscope. The expression of Cav1.2 in LA tissue of the 2 groups was detected by Western blot.RESULTS: Compared with young LA group, dv/dt_max and plateau potential were significantly decreased, APD₃₀ and APD₅₀ were shortened, and APD₉₀ was notably prolongated in aged LA group (P<0.01). The inducibility or duration of atrial arrhythmias was severely increased or prolongated in aged LA group (P<0.01). With voltage clamp model, the density of I_Ca,L in aged LA group was significantly decreased, and current-voltage curve was notably moved upward compared with young LA group. When the clamp potential was +20 mV, the density of I_Ca,L was notably modified from (11.72±1.39) pA/pF in young LA group to (6.08±0.98) pA/pF in aged LA group (P<0.01). Compared with young LA group, the protein level of collagen was significantly increased (P<0.01), and the arrange of atrial myocytes was irregular in LA of rabbits in aged LA group. The atrial myocytes of the LA wall in aged LA group exhibited abnormal ultrastructural alterations, such as karyopyknosis, irregular and swelling mitochondria with the presence of vacuoles, and mild and severe sarcomere degeneration. Compared with those in LA tissues of young rabbits, the expression levels of Cav1.2 in the LA tissues of aged rabbits were severely reduced (P<0.01), and had a significant positive correlation with the reduction of I_Ca,L (r=0.83, P<0.01).CONCLUSION: The pathophysiological characteristics of aged LA are significantly altered, and might contribute to vulnerability and susceptibility of occurrence of atrial fibillation in aged rabbits. The mechanisms might completely attribute to the notable reduction of I_Ca,L, abnormal alterations of ultrastructures and obvious decrease in the expression of Cav1.2 in the aged LA of aged rabbits.     

Effects of schisandrin B on apoptosis of lens epithelial cells treated with H2O2

AIM: To investigate the effects of Schisandrin B (Sch B) on apoptosis of lens epithelial cells (LEC) treated with H₂O₂. METHODS: Eyes in SDrats were excised and lenses were separated under operating microscope in sterilized condition. Lenses were divided randomly into four groups with different treatment: control group, hydrogen peroxide group (H₂O₂), pirenoxine sodium group (PS) and schisandrin Bgroup (Sch B). Lenses were incubated in CO₂ incubator for 24 h with 300 μmol·L^-1 H₂O₂ and with or without 0.5 mmol·L^-1 Sch B. LECaoptosis and apoptosis rate were measured by TUNELmethod. Ultrastructure changes and apoptosis bodies of LECwere observed via transmitted electron microscope. RESULTS: (1) Apoptosis rate in H₂O₂ group (92.0±2.6) was significantly higher than that in control group (3.5±1.8). Apoptosis rate in Sch Bgroup (13.8±3.27) was remarkably lower than that in H₂O₂ group and PSgroup. (2) Ultrastructure observation indicated that apoptosis cells occurred in most LEC in H₂O₂ group and the changes were severe presenting different stages. While a few apoptosis cells were observed in Sch Bgroup, the changes were slight and most of them were in early and middle stages. CONCLUSION: These data indicated that Sch Bsignificantly inhibited apoptosis of LECduring experimental oxidative injury, the effects were stronger than PS.     

Antagonistic effects of cyproheptadine and anisodamine on [Ca2+]i elevation induced by TNFα in endothelial cell strains

AIM: To study the effects of cyproheptadine (Cyp) and anisodamine (Ani) on the changes of intracellular free Ca²⁺ concentration ([Ca²⁺]_i) induced by tumor necrosis factor (TNFα) in single endothelial cells, and to explore the mechanisms of TNFα mediated shock and antishock actions of Cyp and Ani. METHODS: Human umbilical vein endothelial cell strains (ECV304) were seed in 35 mm tissue culture dish with 2 mL DMEM culture medium. The cultured cells were loaded by Fluo-3/AM. The spatial distribution and the dynamic changes of [Ca²⁺]_i in single endothelial cell was determined by laser scanning confocal microscopy (LSCM). RESULTS: [Ca²⁺]_i in single endothelial cell after stimulation of TNFα rapidly increased in a dose-dependent manner and approached the peak value within 60 seconds, afterwards, decreased and kept above the basal level. The confocal scanning image showed that [Ca²⁺]_i elevation was more obvious in nuclear than in cytoplasma, and decreased slowly. Cyp (3×10^-5, 6×10^-5 mol/L) and Ani (2×10^-5, 4×10^-5 mol·L^-1) markedly inhibited TNFα (1.2×10^-9 mol·L^-1)-induced [Ca²⁺]_i elevation. CONCLUSIONS: TNFα markedly induces elevation of [Ca²⁺]_i in single endothelial cell, it may be an important mechanism of TNFα-induced shock and tissue injury. Cyp and Ani obviously suppress TNFα-induced [Ca²⁺]_i elevation, which probably is one of the mechanisms of their antishock effects.