Effects of Astragalus polysacharin on fibroblast proliferation and adhesion between HUVECs and white cells

AIM: To investigate the effects of Astragalus polysacharin(APS) on human fibroblast and human umbilical vein endothelia cell (HUVEC) proliferation, as well as its acts on adhesion between white cells and HUVECs. METHODS: Human fibroblasts from distal and proximal skin away the ulcer were cultured as normal fibroblasts(NF) and wounded fibroblasts(WF). MTT assay was used for detecting cell proliferation, Rose Bengal staining and fluorescence immunohistology assay were used for examining the adhesion of human polymorpho-nuclear cell(PMN) and TPH-1 to HUVECs. RESULTS: 2.44-156 mg/L APS promoted WF proliferation, and 2.44-39 mg/L APS also promoted NF proliferation, but it did not show any proliferating effect on HUVECs. APS inhibited the adhesion of PMN or TPH-1 to HUVECs induced by tumor necrosis factor(TNF). At 25-100 mg/L, it also inhibited both VCAM-1 and ICAM-1 expression in HUVECs induced by TNF. Treatment with APS for 12 h also inhibited CD44 expression in HUVECs. CONCLUSION: APS shows mitogenic activity on both human normal and wounded fibroblasts. It also exerts anti-inflammation effects by inhibiting adhesion molecule expression and adhesion of white cells to HUVECs.     

Effect of ginsenoside Rg1 and Rh1 onthe anti-tumor activity of dendritic cell

AIM: To study the effect of ginsenoside Rg1 and Rh1 on the anti-tumor activity of dendritic cells (DC). METHODS: Effect of Rg1 and Rh1 on the production of IL-12 p40 protein was detected by ELISA, and the IL-12 p40 mRNA level of DC was monitored by RT-PCR. Anti-tumor activity of DC-LPAK was determined by neutral red staining assay. RESULTS: The results of ELISA showed that Rg1 and Rh1 significantly enhanced the production of IL-12 p40 of DC. Rg1 at 1 mg/L and Rh1 at 100 mg/L upregulated the IL-12 p40 mRNA level. Rg1 and Rh1 enhanced the anti-tumor ability of DC, induced lymphokine and PHA activated killer (DC- LPAK) on human papillate tumor cell line. Each dose of Rg1 can obviously accelerate the cytotoxity to L929 at the E∶T ratio of 5∶1(P<0.05,0.01), while only Rh1 10 mg/L enhanced the cytotoxity ability of DC-LPAK (P<0.05). CONCLUSION: Rg1 and Rh1 enhanced the production of IL-12 p40. This effect may be mediated by the increase in the mRNA level. As a result, Rg1 and Rh1 promote the ability of DC to stimulate the cytotoxitic acticity of DC-LPAK.     

Effects of tumor necrosis factor alpha on proliferation and apoptosis of HSCs

AIM:To explore the role of tumor necrosis factor alpha(TNF-α) in the pathogenesis of liver fibrosis.METHODS:The proliferation and apoptosis of hepatic stellate cells (HSCs) in vitro were detected with flow cytometry, electron microscopy and TUNEL.RESULTS:The flow cytometry analysis showed that the cell proliferation index (PI) in the TNF-α(0.5 μg/L, 2.0 μg/L, 8.0 μg/L) groups was evidently lower than that in the control group (P＜0.05). In the cell cycle distribution, the portion of G₀/G₁ phase in the TNF-α groups was significantly higher than that in the control group(P＜0.05), but the portion of S phase in the TNF-α groups was evidently lower than that in the control group(P＜0.05). These indicated that TNF-α interfered with HSCs entrance into S phase from G₀/G₁ phase whereupon the proliferation of HSCs was inhibited. The apoptotic rate in the TNF-α groups was evidently higher than that in the control group(P＜0.05). The gene expression of bcl-2 and bax was also detected with flow cytometry. The expression of bcl-2 in the TNF-α groups was evidently lower than that in the control group(P＜0.05), but the expression of bax in the TNF-α groups was significantly higher than that in the control group(P＜0.05). TUNEL analysis showed the apoptotic rate of HSCs in the TNF-α(2.0 μg/L) group was 18.7%±2.5% compared with 5.3%±1.2% in the control group(P＜0.05).CONCLUSIONS:TNF-α interfered with HSCs entrance into S phase from G₀/G₁ phase whereupon the proliferation of HSCs was inhibited. TNF-α down-regulated bcl-2 gene expression and up-regulated bax gene expression whereupon the apoptosis of HSCs was induced.     

Protective effect of astragalus polysaccharide on mitochondria and lysosome in H2O2-stressed skin fibroblasts

AIM: To verify the protection of astragalus polysaccharide (APS) on H2O2-stressed skin fibroblasts. METHODS: A model of acute H2O2 stress in primary skin fibroblast was used at concentration of 0.5 mmol/L by 30 min incubation. Dose responses of APS on cell survival was measured by MTT, cell death was evaluated by DAPI, and effect of APS on mitochondria, mitochondrial membrane potential and lysosome stabilization were measured by confocal microscopy. RESULTS: APS improved cell survival in a dose-dependent manner, starting at 0.5 mg/L and with a maximum at 1 mg/L. Moreover, APS inhibited mitochondrial membrane potential collapse, protected mitochondrial morphology and stablized lysosomal membrane. CONCLUSION: The results suggest the existence, at the mitochondria-lysosome level, of a new pathway of apoptotic regulation by APS. This might constitute a new therapeutic target where oxidative stress and lysosomal impairment are involved.     

Effects of testosterone on the fibrinolysis activity of HUVEC and its mechanism

AIM: To investigate the effect of testosterone with varied concentrations on the fibrinolysis activity of HUVEC and its mechanism.METHODS: Human umbilical vein endothelial cells (HUVEC) were cultured as recommended. After confluence, the cultures were treated with testosterone(3 ×10-10, 3×10-9, 3×10-8,3×10-6, 3×10-5 mol/L) , and the control confluent cells were cultured in the same medium without steroid. MTT experiment was repeated for 72 hours to investigate each groups’ cell proliferation. The tPA and PAI-1 antigen levels were assayed with ELISA Kits. Then with HUVEC incubated in androgen receptor antagonist (flutamide) 3 hours previously, the experiment was repeated. RESULTS: Testosterone at physiologic or lower concentrations (3 ×10-10 to 3×10-8 mol/L ) stimulated the secretion of tPA by HUVEC (P＜0.01). However, tPA levels markedly reduced at larger doses (3 ×10-6 to 3×10-5 mol/L). On the other hand， PAI-1 antigen levels decreased significantly at the testosterone concentrations ranging from 3 ×10-10 to 3×10-5 mol/L (P＜0.05). Flutamide attenuated the testosterone’s effects (P＜0.05). CONCLUSIONS: The results indicated that testosterone at physiologically relevant concentrations decreased PAI-1, while increased tPA levels via the androgen receptor, which suggested that testosterone may have beneficial effects on preventing thrombotic diseases.     

Effect of antifibrotic herb serum on rat hepatic stellate cell proliferation and collagen synthesis

AIM: To investigate the effect of Chinese herbs, Ganxianfang(GXF), on rat hepatic stellate cells (HSC) proliferation and collagen synthesis. METHODS: Two types of herb serum, portal venous serum and circumferential venous serum, were prepared from rats infused intragastrically with 16, 8, 4 times adult dose of GXF decoction. HSC isolated from rat liver were processed with the above sera in vitro. Then we mensurated the radioactivity of HSC admixed with [[³H]H]proline and [[³H]H]thymine to judge the effect on proliferation and collagen synthesis of HSC. RESULTS: Both two types of serum collected 0.5, 1, 2 h after intragastrical infusion inhibited HSC proliferation (P＜0.05), and the serum collected 1 h after intragastrical infusion had the strongest effect (P＜0.05). Portal serum decreasea collagen synthesis (P＜0.05), but circumferential serum had no effect (P＞0.05). CONCLUSION: Inhibition of HSC proliferation and decrease of collagen synthesis may contribute to the GXF antifibrotic action.     

Mechanism responsible for inhibitory effect of recombinant rat augmenter of liver regeneration on renal turbular epithelial cell apoptosis by gentamycin

AIM: To investigate the effects of recombinant rat augmenter of liver regeneration (rrALR) on apoptosis of renal tubular cells (NRK-52E cells) induced by gentamycin sulfate (GM). METHODS: The cultured NRK-52E cells were divided into four groups: normal control cells, cells with GM (GM 1.6 g/L) or GM and rrALR (15 mg/L or 25 mg/L) treatments. The apoptosis of cultured cells were assessed at 24 h, 48 h by AO/EB staining, DNA agarose gel electrophoresis analysis and flow cytometry using Annexin V-FITC and propidium iodide (PI) staining. The protein and mRNA expressions of Bcl-2 and Bax were detected by Western blotting and RT-PCR, respectively. RESULTS: (1) rrALR inhibited NRK-52E cells apoptosis induced by GM (P＜0.05). (2) rrALR promoted the expression of Bcl-2 protein and mRNA, but inhibited the Bax protein and mRNA expression (P＜0.05) in cultured NRK-52E cells in a dose-dependent manner. The value of Bcl-2/Bax increased. CONCLUSION: rrALR inhibits renal tubular epithelial cell apoptosis and ameliorates cell injury induced by nephrotoxic drug GM presumably via the regulation of Bcl-2 and Bax protein and mRNA expressions.     

The adaptive response of low dose sodium nitrite pretreatment on cultured CHL cells to DNA damage caused by high dose sodium nitrite

AIM: To study if low dose NaNO₂ can induce the adaptive response of cultured Chinese hamster lung cells（CHL cells）to DNA damage. METHODS: Single cell gel electrophoresis technique was used to detect the DNA damage in CHL cells exposed to NaNO₂ at different concentrations. CHL cells were pretreated with NaNO₂ of concentrations of 0.01 mg/L, 0.1 mg/L and 1 mg/L respectively. And the adaptive response to the toxicity of 1g/L NaNO₂ was observed. The activity of polyADP- ribose polymerase (PARP-1) of CHL cells was inhibited with 3-aminobenzamide(3AB) before or after pretreated with low dose of NaNO₂. And the changes of the adaptive response were observed. RESULTS: The rate of tailing cells was 7.87% when the cells were exposed to 1 g/L NaNO₂ without pretreatment with low dose NaNO₂. An extremely remarkable statistics significance (P＜0.01) was observed when compared the difference to control group. NaNO₂ of 0.01 mg/L and 0.1 mg/L could induce the adaptive response of cultured CHL cells to DNA damage caused by 1 g/L NaNO₂. The rate of tailing cells was 3.55% and 1.06％ respectively, which was much lower than that of no-pretreatment group(P＜0.05；P＜0.01). But the rate of tailing cells was 6.09% when the cells were exposed to 1 g/L NaNO₂ with pretreatment of 1 mg /L NaNO₂, which had no significant difference compared with the rate of tailing cells in control group (P＞0.05). The adaptive response could be blocked when the activity of PARP-1 was inhibited with 3AB before the low dose pretreatment, but could not be blocked when the activity of PARP-1 was inhibited after low dose NaNO₂ pretreatment 6 h. CONCLUSION: NaNO₂ of concentration that equals to or lowers than 0.1 mg/L can induce the adaptive response of cultured CHL cells to DNA damage caused by high dose NaNO₂ through PARP-1 activation. And the dose of NaNO2 that can induce adaptive response might not cause the DNA damage.     

不同基因型梨叶片离体培养和植株再生

 以‘巴梨’、‘身不知’和‘早酥’梨试管苗叶片为外植体, 对不定芽进行了诱导、增殖和生根。重点探讨了基本培养基、植物生长调节剂配比、AgNO₃ 不同浓度和NH4⁺-N与NO3^--N比例对不定芽再生的影响。结果表明, MS + TDZ 0.5 mg/L + IBA 0.1 mg/L为‘巴梨’叶片不定芽发生的最佳培养基。在QL + TDZ 1.0 mg/L +NAA 0.1 mg/L培养基上, 暗培养3周后转光下培养, ‘身不知’和‘早酥’分别获得89.6%、81.2%的不定芽再生率和3.45、3.73的平均再生芽数; 对‘巴梨’和‘早酥’不定芽再生有效促进的AgNO₃ 浓度范围为0.1～0.5 mg/L, 0.1～4.0 mg/L的AgNO₃、1∶2～7的NH₄⁺-N∶NO ₃^--N和21.50mmol/L的K+对‘身不知’叶片再生均有促进作用, 缺乏NH₄⁺-N不利于不定芽的再生; 转移到MS+BA1.0 mg/L + IBA 0.1 mg/L培养基上能快速增殖; 在MS、1 /4MS + IBA 1.0～2.5 mg/L +蔗糖5～15 g/L +活性炭0.5～1.0 g/L上获得了不同程度的生根苗。     

Effects of insulin on cardiac fibroblast proliferation and cardiac myocyte hypertrophy

AIM:To study the effect of insulin on proliferation and hypertrophy of cardiac myocytes and its role in the induction of cardiac hypertrophy. METHODS:1. The neonatal rat cardiac myocytes and cardiac fibroblasts were cultured respectively and identified with light microscopy, electron microscopy and immunocytochemistry. 2. Cell proliferation was measured with cell number, metabolic activity and DNA synthesis (with WST-1, BrdU enzyme-linked immunosorbent assay ) and the percentage of S+G₂+M in cell cycle (by flow cytometry ). 3.Cell hypertrophy was evaluated by cell protein content (Coomassie Briliant Blue's method). RESULTS:1. The cultured cells showed the characteristic of cardiac myocytes and cardiac fibroblasts, respectively. 2. After being treated with insulin, the cell number, absorbance of BrdU incorporation and WST-1 cleavage products and the percentage of S+G₂+M of cardiac fibroblasts increased significantly (P＜0.01 orP＜0.05), while the above parameters of cardiac myocytes remained unchanged (P＞0.05). 3. Protein content of cardiac myocytes increased significantly in a dose-dependent manner (P＜0.01 orP＜0.05) in insulin treated groups (10^-10 mol/L-10^-7 mol/L). CONCLUSION:Insulin promoted cardiac fibroblast proliferation and increased myocytes protein content(induced myocyte hypertrophy)in vitroand may play an important role in pathogenesis of cardiac hypertrophyin vivo.     

樱桃砧木‘大青叶’茎段离体培养与快繁技术

以引种栽培的樱桃砧木‘大青叶’茎段为试材,探讨基本培养基和植物生长调节剂对茎段腋芽萌发、生长与增殖的效应,筛选离体培养的最适培养基。结果表明：MS＋6－BA0．5mg／L＋zT 0．1mg／L＋蔗糖20g／L＋琼脂0．6mg／L、MS＋6－BA1．0mg／L＋ZT 0．1g／L＋蔗糖20g／L为丛生芽诱导及分化增殖的最适培养基;1／2MS＋IBA 0．7mg／L＋NAA 0．2mg／L＋琼脂0．6mg／L为生根诱导的最佳培养基配方。     

Effects of oxidized low density lipoprotein and vitamin E on the levels of IL-6,IL-8 and TNF-α in cultured human umbilical vein endothelial cells

AIM:To investigate effects of OX-LDL and VitE on the levels of IL-6,IL-8 and TNF-α in human umbilical vein endothelial cells(HUVEC).METHODS: Human umbilical vein endothelial cells were obtained by in vitro culture. HUVEC treated with or without Vit E was incubated with OX-LDL, and the levels of IL-6, IL-8 and TNF-α were determined by enzyme-linked immunosorbent assy technique. RESULTS:50 μg/L,100 μg/L, 200 μg/L OX-LDL induced the release of IL-6,IL-8 and TNF-α by HUVEC in a dose-dependent manner. Compared with the control group , the levels of IL-6 and IL-8 were significantly increased at 6-12 h of stimulation with OX-LDL . Maximal levels of IL-6 and IL-8 occurred after 24-36 h, reaching a plateau maintained for at least 48 h. TNF-α rose after 2-6 h in HUVEC, and reached a maximum after 12 h. In contrast to IL-6 and IL-8, TNF-α declined after 48 h. However, when VitE (50 mg/L,100 mg/L,200 mg/L)was added, it can significant inhibited the release of IL-6, IL-8 and TNF-α in a dose-dependent manner, and after 48 h these cytokines have no diference between OX-LDL+VitE groups and OX-LDL groups. CONCLUSION: OX-LDL can obviously stimulate the production of IL-6,IL-8 and TNF-α in vascular endothelial cells, which can significantly be inhibited by VitE in a short time.     

Effects of recombinant human CD40 ligand on biological behavior of ovarian cancer SKOV3 cells in vitro

AIM: To investigate the effect of recombinated human CD40 ligand (rhCD40L) on the biological behavior of ovarian cancer SKOV3 cell line in vitro.METHODS: After the SKOV3 cells were incubated with different concentrations of rhCD40L for various times, the cell proliferation was determined by MTT assay.The expression of the co-stimulatory molecules or adhesion molecules on SKOV3 cells and the changes of tumor necrosis factor receptor associated factor (TRAFs) inside the cells were measured by flow cytometry and direct immunofluorescence.Annexin V and PI dual color label assay were used to detect cell apoptosis or death in culture contained with rhCD40L.RT-PCR assay was employed to determine the change of apoptosis related gene c-myc, bcl-2 and bcl-xl expression in SKOV3 cells.RESULTS: rhCD40L inhibited proliferation of SKOV3 cells at concentration of 100 μg/L (0.65±0.10 vs 0.81±0.05) and reached a peak at concentration of 10 mg/L (0.13±0.12 vs 0.83±0.15, P＜0.01).The inhibitory effects showed a dose dependent manner.Cell cycle analysis showed that cell division was blocked in G1 phase.Increasing proportion of apoptosis of SKOV3 cells was related to up-regulation of CD95 expression (42.4% vs 59.2%, P＜0.05) and down-regulation of anti-apoptosis genes such as bcl-2 and bcl-xl expressions after incubation with rhCD40L.TRAF 2, 5 and 6 expressed highly in SKOV3 cells.The expression of TRAF 2 (81.3%±9.2% vs 50.4%±5.3%，P＜0.05), TRAF5 (47.2%±7.2% vs 7.2%±2.1%, P＜0.01) and TRAF6 (44.5%±6.3% vs 5.1%±1.1%, P＜0.01) was down-regulated and expression of TRAF 3 (25.2%±6.2% vs 68.8%±5.3%, P＜0.01) was up-regulated after co-culture with rhCD40L, but there was no effects found on the expression of TRAF 1 (4.3%±1.2% vs 5.1%±1.4%) and TRAF4 (7.4%±1.2% vs 8.1%±1.4%).CONCLUSION: By down-regulating expression of bcl-2, bcl-xl and changing expression profile of TRAF, rhCD40L inhibits the growth of SKOV3 cells by blocking the cell cycle progress in G1 and promotes the cells to apoptosis.     

Effects of nicotine on activation of PMNs and the ICAM-1 mRNA expression of HUVEC

AIM: To investigate the effects of nicotine on activation of PMNs, adhesion of PMNs-HUVEC and expression of ICAM-1 mRNA in HUVEC. METHODS: Activation of PMNs was measured by detecting the activity of β-glucuronidase and lysozym of PMNs. Adhesion of PMNs and HUVEC was observed. Northern blot was conducted for quantitating ICAM-1 mRNA. RESULTS: Nicotine could increase the activity of β-g [(8.76± 1.01)μg/10⁷·h vs(14.87±2.00)μg/10⁷·h,P<0.05]and Lysozym [(20.0±1.5)μg/10⁷·h vs(36.5±4.4)μg/10⁷·h,P<0.05], and also could promote adhesion of PMNs-HUVEC(38.5±9.8 vs 61.0±4.4,P＜0.05). The expression of ICAM-1 mRNA was induced by nicotine in dose-dependent fashion (10^-5-10^-3mol/L).After a 2 h treatment of HUVEC with nicontine(10^-4mol/L), the level of ICAM-1 mRNA is above the control(1.23 vs 1.63) and the highest level (2.03) is at a 12 h treatment. 764-3 can obviously counteract the above effect of nicotine. CONCLUSIONS: Nicotine could activate PMNs, enhance adhesion of PMNs-HUVEC and increase the expression of ICAM-1 mRNA in HUVEC.     

Modulatory roles of muscarinic cholinergic receptor 3 in proliferation and migration of small cell lung cancer

AIM: To study the expression and roles of muscarinic cholinergic receptor 3 (M3R) in human small cell lung cancer (SCLC) cells. METHODS: Human SCLC cell lines SBC3 and H82 were cultured in vitro. RT-PCR and Western blotting were used to investigate the expression of M3R. MTT assay and Boyden chamber assay were carried out to determine the roles of cholinergic receptor agonist acetylcholine iodide (ACh) and M3R antagonist 4-diphenylacetoxy-N-methylpiperidine methiodide (4-DAMP) in the proliferation and migration of SBC3 cells. RESULTS: M3R was expressed in SBC3 and H82 cells. The relative protein expression of M3R normalized with β-actin in SBC3 was 2.65-fold higher than that in H82. ACh stimulated SBC3 cell proliferation in a dose-dependent manner. Treatment with ACh at concentrations of 10^-4 and 10^-3 mol/L significantly stimulated SBC3 cell growth at 48 h and 72 h (P＜0.01). SBC3 cell proliferation induced by ACh was inhibited by 4-DAMP in a dose-dependent manner. Pretreatment of the cells with 10^-5 mol/L 4-DAMP suppressed the effect of ACh at 48 h (P＜0.05). Pretreatment with 4-DAMP at concentrations of 10^-6, 10^-7 (P＜0.05) and 10^-5 mol/L (P＜0.01) inhibited the effect of ACh at 72 h. Treatment with 10^-5 or 10^-6 mol/L 4-DAMP alone inhibited the cell proliferation at 48 h (P＜0.01) and the inhibitory effect of 4-DAMP at concentration of 10^-5 mol/L was stronger than that of 4-DAMP at concentration of 10^-6 mol/L at 72 h. ACh increased the cell migration towards fibronectin (Fn) in a dose-dependent manner and ACh at concentration of 10^-4 mol/L enhanced the cell migration by about 3 folds. The cell migration stimulated by 10^-4 mol/L ACh was almost completely blocked by pretreatment with 4-DAMP at concentration of 10^-6 or 10^-5 mol/L (P＜0.01). CONCLUSION: M3R is expressed in human SCLC cells. The M3R antagonist inhibits SBC3 cell proliferation and migration.     

中国园艺学会第七届青年学术讨论会

中国园艺学会第九届第8次常务理事扩大会决定,“中国园艺学会第七届青年学术讨论会”由山东农业大学园艺科学与工程学院和山东省园艺学会承办,将于2006年7月或8月在山东泰安举行。会议交流主题:(1)园艺作物种质资源、遗传育种与生物技术;(2)园艺作物有机、无公害及标准化安全生     

Metallothionein inhibits homocysteine-induced proliferation of rat vascular smooth muscle cells

AIM:To investigate the effect of metallothionein(MT) on proliferation of rat vascular smooth muscle cells (VSMCs) stimulated by homocysteine and its mechanism. METHODS:VSMCs proliferation was measured by [³-H]-TdR incorporation, mitogen-activated protein kinase(MAPK)activity were determined by immunoprecipitation method, the intracellular contents of MT and malondialdehyde (MDA)were assayed by -hemoglobin saturation method and TBA reaction, respectively, and lactate dehydrogenase (LDH) leakage was measured by NADH oxidation. RESULTS:Hcy(10^-6-10^-4 mmol/L) stimulated [³-H]-TdR incorporation by the VSMCs in a concentration-dependent manner. Compared with control, [³-H]-TdR incorporation in VSMCs treated with 0.1 mmol/L Hcy was increased by 4.2 fold (P＜0.01). Meanwhile, Hcy enhanced MAPK activity, MDA formation and LDH release (P＜0.01)in a concentration-dependent manner. Treatment of VSMCs with MT alone did not change above parameters, compared with control. However, MT (10^-6-10^-4 mol/L)attenuated significantly Hcy-stimulated proliferation of VSMCs (P＜0.01)in a concentration-dependent manner. And MT inhibited obviously Hcy-induced activation of MAPK activity, MDA formation and LDH release. Preincubation of VSMCs with 0.5 mmol/L ZnCl₂ for 6 h induced an increase cellular MT content by 5.7-fold (P＜0.01). The MT-overexpressed VSMCs resisted Hcy-stimulating action on MAPK activity, MDA formation and LDH leakage (P＜0.01). CONCLUSION:These results show that MT has an inhibitory effect on Hcy-induced VSMCs proliferation, and that MT could inhibit Hcy-stimulated MAPK activity and lipid peroxidation.     

卷丹转基因体系构建及岷江百合LrCCoAOMT的导入

以卷丹（Lilium lancifolium Thunb.）无菌小鳞片为试材，通过切片处理、优化农杆菌侵染浓度与时间以及重悬液和共培养基成分，构建农杆菌介导的高效遗传转化体系。结果表明，MS + 1.5 mg ? L-1 6-BA + 0.5 mg ? L-1 NAA + 30 g ? L-1蔗糖是切片芽分化的最佳培养基，添加100 mg ? L-1抗坏血酸能有效抑制褐化并促进不定芽增殖。MS + 2.0 mg ? L-1 NAA + 30 g ? L-1蔗糖是生根诱导的最佳培养基。抗生素敏感性测试发现，培养基中添加Kan 100 mg ? L-1或Hyg 75 mg ? L-1 结合Cef 400 mg ? L-1适宜抗性筛选。GUS染色分析表明，以去除大量元素的改良MS + 100 μmol ? L-1 AS和去除大量元素的改良MS + 1.0 mg ? L-1 6-BA + 1.0 mg ? L-1 NAA + 100 μmol ? L-1 AS为重悬液和共培养基，将切片在农杆菌菌液浓度OD600为0.4，侵染15 min，获得81.72% 瞬时转化率和25.2% 稳定遗传转化率。将岷江百合LrCCoAOMT转化卷丹，分子检测和GUS染色分析表明已获得转基因阳性株系。     

采用L_9(3~4)正交设计方法筛选血耳菌丝体液体培养基

采用正交设计方法以血耳Tremella sanguinea菌丝体的生物量为测量指标,对血耳菌丝体液体培养基进行了筛选,结果表明血耳菌丝体液体培养的最适培养基为蔗糖7.5g·L-1、麦芽糖7.5g·L-1、麦麸7.5g·L-1、牛肉膏3.5g·L-1、MgSO4·7H2O0.5g·L-1、KH2PO41.5g·L-1、VBl4mg·L-1,pH自然。28℃培养96h,其生物量可达4.29g·L-1。