灵雷菌质的急性毒性实验研究

研究双向性固体发酵后灵雷菌质的急性毒性。用双向性固体发酵技术获得灵雷菌质,用Bliss法计算小鼠的半数致死量（LD50）。结果表明各灵雷菌质组的LD50均高于雷公藤生药组,尤以3G30组最高。双向性固体发酵技术可降低雷公藤的急性毒性。     

灵丹菌质双向发酵工艺及药效品质研究

采用双向固体发酵工程技术,以灵芝为发酵菌株,在含有不同比例丹参边角料的培养基中进行发酵,观察了不同培养基中菌丝的生长情况,测定了发酵产物（灵丹菌质）中多糖、蛋白质、三萜、丹参酮的含量变化,并研究了其提取物对小鼠活血化瘀功效的影响。结果表明灵丹菌质的最佳发酵组合为组合4,发酵40d,多糖含量26.48μg·g-1,蛋白质含量为397.24μg·g-1,灵芝酸三萜含量为4.92%。药效学实验显示,灵丹菌质组合4具有活血化瘀的药效,优于丹参药材。     

基于牛蒡根的灵芝固体发酵菌质化学成分的初步分析

以牛蒡根为发酵基质,初步探究灵芝固体发酵所获菌质中多种化学成分的变化。利用可见分光光度计、高效液相色谱仪及定氮仪等测定了牛蒡根原药材和牛蒡根灵芝菌质中多糖、总三萜酸、总黄酮、核苷及蛋白质含量,并进行HPLC指纹图谱分析。牛蒡根经灵芝菌丝体发酵后,多糖含量由9.94%降低至2.76%,总三萜酸含量从无升高至0.14 mg·g^-1,总黄酮含量由3.47%降低至0.34%,4种核苷总量由861.05μg·g^-1升高到1 113.49μg·g^-1,且新合成了腺苷,蛋白质含量由13.38%升高至17.28%。HPLC指纹图谱显示牛蒡根在灵芝发酵前后化学成分差异较大。灵芝固体发酵使牛蒡根多糖和黄酮含量降低,核苷和蛋白质含量升高,并新产生三萜酸和腺苷,具有明显的生物转化作用。     

灵雷菌质提取物体内抗肿瘤作用的实验研究

采用小鼠肉瘤(S180)移植性肿瘤模型,观察灵雷菌质乙酸乙酯部位、正丁醇部位、石油醚部位和水部位提取物的肿瘤抑制作用,并测定小鼠的胸腺指数及脾脏指数。结果表明所有的提取物都呈现出明显的抑瘤效果,抑瘤率均大于35%,其中部分正丁醇部位的抑瘤率达到了68.88%。抑瘤效果由弱至强的顺序为石油醚部位<水部位<乙酸乙酯部位<正丁醇部位,而胸腺指数与脾脏指数则显示只有乙酸乙酯部位组有较明显的免疫抑制效果。双向性固体发酵可降低雷公藤的毒性,尤其正丁醇部位和水部位的毒性可能较弱。     

猴头人参花发酵菌质化学成分及抗胃溃疡作用研究

目的：研究小刺猴头菌与人参花共发酵菌质的化学成分变化以及对小鼠胃溃疡的治疗作用。方法：将小刺猴头菌接种于人参花固体培养基中共发酵,获得猴头人参花菌质。利用紫外可见分光光度法测定发酵前后人参花多糖和人参总皂苷的含量变化。建立无水乙醇致小鼠胃溃疡模型,观察猴头人参花共发酵前后人参花、猴头人参花发酵物70％乙醇和水提取物对小鼠的胃溃疡面积以及对溃疡抑制率的影响。结果：小刺猴头人参花菌质多糖含量〉人参花高压灭菌后多糖含量〉人参花多糖含量。人参花高压灭菌后总皂苷含量〉人参花总皂苷含量〉小刺猴头人参花菌质总皂苷含量。人参花、高压灭菌人参花、猴头人参花发酵菌质的70％乙醇提取物对抑制小鼠胃溃疡有一定疗效。结论：人参花在高压灭菌的过程中增加了多糖和总皂苷的含量,和猴头菌共发酵会使多糖含量增加,但人参总皂苷含量没有显著变化。猴头人参花发酵菌质比单独使用人参花治疗胃溃疡的疗效更为显著。     

基于响应面的蛹虫草菌质固体发酵工艺条件优化

优化蛹虫草固体发酵菌质工艺以缩短发酵周期,提高有效成分含量,有效节约成本。通过单因素筛选和响应面优化法对碳源、氮源、料液比、小麦装料量、接种量、培养时间和腺苷浓度等培养条件进行优化。结果显示,蛹虫草固体发酵菌质最佳培养方案的小麦装料量为15 g,料液比为1.0∶1.3,接种量为2 mL,培养时间为17 d,营养液碳源为可溶性淀粉、氮源为黄豆粉,腺苷的质量浓度为76 g·L^-1。此条件下虫草素含量为3.718mg·g^-1,与初始配方相比,含量提高了4.47倍。工艺优化后的蛹虫草菌质固体发酵周期短,有效成分含量提高,研究结果为后期发酵菌质的开发利用奠定了基础。     

药用真菌"双向发酵"的起源、发展及其优势与潜力

笔者上世纪80年代末提出用药用真菌发酵药材的设想,实验证实在农副产品构成的营养基质内掺入适量经特选的含有活性成分的中药材作为药性基质,致使发酵具有双向性,其产物由药用菌质(如槐耳菌质)转为药性菌质(如槐芪菌质○F),可以显著增强其免疫活性,提高药效与疗效,这是现代“双向发酵”工程的起源,此后因理论与技术发展,使它被认为是生物技术的新起点,中药新药研制的新途径,各种菌质必将成为中药内菌物药的重要组成。较深入和更扩大的研究认为,它有增效、扩用、解毒持效等作用可发展为用中药二次开发和兽药领域。与其它有关生物技术比较,在现阶段有一定的优势与潜力。“双向发酵”研发道路宽广,前景良好,也还存在需要研究解决的问题,如怎样以中医药理论指导构成发酵组合和对各种菌质的科学定性等,尤其需要从基质发酵为菌质的大规模生产的机械化、自动化设备。     

食(药)用菌抗疲劳作用实验研究

将香菇和灰树花2种食（药）用菌的菌种分别接种到灵芝渣和云芝渣上（提取下脚料）,进行固体发酵,然后与全灵芝破壁孢子粉分别采用超声波水提法和醇提法制备提取物,通过小鼠灌胃20d-30d后进行负重游泳实验,初步评价和验证上述提取物的抗疲劳作用。试验结果表明：香菇固体发酵物的醇提液、灰树花固体发酵物及全灵芝破壁孢子粉水提物组小鼠的负重游泳时间比对照组显著延长（P〈0.05）;提示上述提取物具有抗疲劳作用。     

真菌药用"固体发酵"上游工程建设基本研究方法

真菌药用"固体发酵"是中药传统范畴内仅有的,也是历史上诞生最早的一类生物制药技术,并已发展走向"固体发酵系列工程".固体发酵的上游工程建设,特别是发酵组合的优选、发酵工艺的制定等,对决定所产"菌质"的性效有重要影响.据研发槐耳、槐芪、灵雷3种"菌质"的经验、体会,提出几项基本研究方法,供业内各方参用、改进,以使达规范化.     

栽培与野生桦褐孔菌菌核的生药学比较

采用显微鉴定和理化检识的方法对栽培桦褐孔菌和野生桦褐孔菌菌核进行生药学的比较。结果显示显微鉴定中栽培与野生桦褐孔菌菌核差别在于前者菌丝松散,分枝较少;后者相反。理化检识方面,薄层层析比较得到二者均含有白桦酯醇;GC-MS联用分析鉴定出栽培桦褐孔菌菌核石油醚提取物中8个化合物,占总量的70.31%;野生桦褐孔菌菌核石油醚提取物中4个化合物,占总量的16.04%,并且二者没有共同的化合物;高效液相色谱分析出栽培桦褐孔菌的粗水提取物和野生桦褐孔菌菌核的粗水提物在相同条件下,栽培桦褐孔菌菌核粗水提取物明显的峰有2个,保留时间分别为85.026 min和107.920 min;野生桦褐孔菌菌核粗水提物明显的峰有4个,保留时间分别为76.310 min、77.130 min、87.546 min和97.423 min。可以看出,相同条件下二者比较,并未发现相同的成分。     

Relationship between renal hypertension and cardiac α1-adrenoceptor autoantibodies in rats

AIM: To examine the autoantibody against α₁-adrenoceptor and its biologic activities during the development of renal hypertension. METHODS: Renal hypertension of rat was achieved by clipped renal artery, the titre of autoantibody to α₁-adrenoceptor was detected using ELISA immunoassay. Furthermore, the biological offects of these autoantibodies on cultured cardiomyocytes were also examined. RESULTS: After two weeks of clipping renal arteries, both the frequency of occurrence and the titre of autoantibodies to cardiac α₁-adrenergic receptor were significantly increased as compared with the control of pre-treatment. The increased autoantibodies lasted for several weeks and then automatically decreased gradually to the pre-clipping level at 12 weeks. The biological effects of these autoantibodies displayed an "agonistic-like" activities on the beating frequency of cultured neonatal cardiomyocytes. CONCLUSION: Autoantibodies against α₁-adrenoceptor may play a role in the elevation of peripheral vascular resistance and in the development of cardiac hypertrophy in rats with renal hypertension.     

Effects of radiation from 188Re on smooth muscle cell proliferation

AIM: Although endovascular radiotherapy inhibits neointimal hyperplasia, the exact alterations induced by β-particles irradiation remain to be elucidated. The objective of this study was to investigate the ability and the cellular mechanism of local β^-particles emission from ¹⁸⁸Re to inhibit vascular smooth muscle cells (SMCs). METHODS: The SMCs in vitro were irradiated by ¹⁸⁸Re with single doses of 2.6 Gy-25.8 Gy. The effects of β-particles on SMCs, such as effective irradiate doses, the period of inhibition for SMCs proliferation, the changes of cell proliferation rate and DNA synthesis rate, cell cycle progression and related gene expression, were investigated by cell count, [³H]-TdR incorporation, cell cycle progression analysis, cell viability and immunocytochemistry, respectivecy. RESULTS: β-particles irradiation with dose of 5.2 Gy could inhibit significantly SMCs proliferation. At dose of 20.6 Gy DNA synthesis inhibitory rate was 92%, SMCs proliferation rate was only 3%. Renoval of ¹⁸⁸Re did not abolish the inhibitory effects of β-particles on SMCs proliferation. The expression of P53 was up regulation and PCNA was down regulation after irradiation. CONCLUSION: β-particles from ¹⁸⁸ Re was significantly effective and permanent in inhibiting SMCs proliferation, and inhibitory effect was in dose-dependet manner ED50was 5 Gy, the best dose to inhibit SMCs proliferation was 20 Gy. β-particles irradiation induced SMCs to occur G₀/G₁ arrest, damaged the ability of SMCs reproliferation and led to cell clonogenic death. P53 and PCNA had regulatiory effects on SMCs proliferation after β-particles irradiation.     

Influence of Mineral Nutrients on Strawberry Fruit Quality and Their Accumulation in Plant Organs

Abstract

This review is based partly on complete articles and partly on abstracts. Three of the 60 articles deal with the total uptake of elements in strawberry plant organs in two different strawberry production systems, both considered as optimal concerning amount and balance of elements. The effect on fruit quality may be dramatic if the level of a particular element is outside this range, but there may also be effects initiated by differences within the optimal range of elements. Most articles refer to product oriented quality, but some focus on consumer oriented quality, as discussed by Shewfelt (1999). The discussion here is on a general basis, so one should keep in mind that there are cultivar differences and that specification of nutrition ideally should mirror the needs of a single cultivar, or a group of cultivars with similar requirements. Also, to get a complete understanding of the subject future reviews should embrace a broader access of information including the effect on plant development of individual elements, such as the role of calcium in fruit firmness and its importance in cell wall structure. However, the intention here is to narrow the information to results that suggest a direct connection between nutrient uptake and fruit quality.     

Effect of L-Arg on plasma content of endothelin and expression of c-fos mRNA in the left ventricle of rats with renovascular hypertensive hypertrophy

AIM:To study the effect of L-Arg on plasma content of endothelin (ET) and the expression of proto-oncogene c-fos mRNA in the left ventricle of rats with renovascular hypertensive hypertrophy. METHODS: The level of c-fos mRNA were measured by in situ hybridization. The ET in plasma were measured by radioimmunoassay. RESULTS:After eight weeks of treatment with L-Arg, the expression of c-fos decreased markedly (P＜0.01). The ET content in plasma also decreased significantly by L-Arg(P＜0.01).CONCLUSION: Plasma ET content and the expression of c-fos in the left ventricle of rats with renovascular hypertensive hypertrophy could be decreased by L-Arg administration.     

Mixed-severity fire regime in a high-elevation forest of Grand Canyon,Arizona, USA

Fire regime characteristics of high-elevation forests on the North Rim of the Grand Canyon, Arizona, were reconstructed from fire scar analysis, remote sensing, tree age, and forest structure measurements, a first attempt at detailed reconstruction of the transition from surface to stand-replacing fire patterns in the Southwest. Tree densities and fire-/non-fire-initiated groups were highly mixed over the landscape, so distinct fire-created stands could not be delineated from satellite imagery or the oldest available aerial photos. Surface fires were common from 1700 to 1879 in the 4,400 ha site, especially on S and W aspects. Fire dates frequently coincided with fire dates measured at study sites at lower elevation, suggesting that pre-1880 fire sizes may have been very large. Large fires, those scarring 25% or more of the sample trees, were relatively infrequent, averaging 31 years between burns. Four of the five major regional fire years occurred in the 1700s, followed by a 94-year gap until 1879. Fires typically occurred in significantly dry years (Palmer Drought Stress Index), with severe drought in major regional fire years. Currently the forest is predominantly spruce-fir, mixed conifer, and aspen. In contrast, dendroecological reconstruction of past forest structure showed that the forest in 1880 was very open, corresponding closely with historical (1910) accounts of severe fires leaving partially denuded landscapes. Age structure and species composition were used to classify sampling points into fire-initiated and non-fire-initiated groups. Tree groups on nearly 60% of the plots were fire-initiated; the oldest such groups appeared to have originated after severe fires in 1782 or 1785. In 1880, all fire-initiated groups were less than 100 years old and nearly 25% of the groups were less than 20 years old. Non-fire-initiated groups were significantly older (oldest 262 years in 1880), dominated by ponderosa pine, Douglas-fir, or white fir, and occurred preferentially on S and W slopes. The mixed-severity fire regime, transitioning from lower-elevation surface fires to mixed surface and stand-replacing fire at higher elevations, appeared not to have been stable over the temporal and spatial scales of this study. Information about historical fire regime and forest structure is valuable for managers but the information is probably less specific and stable for high-elevation forests than for low-elevation ponderosa pine forests.This revised version was published online in May 2005 with corrections to the Cover Date.     

Compositional and Functional Properties of Saskatoon Berry and Blueberry

Abstract

Saskatoon berry (Amelanchier alnifolia Nutt., Rosaceae) and blueberry (Vaccinium corymbosum L., Ericaceae) are substantially equivalent in all characteristics that are important to the consumer, including fruit color, shape, size, nutrition, texture, and uses. In addition, both fruits are native to North America and they have practically identical historical uses and known health benefits. Their composition, processing, nutritional value and metabolism, intended uses, and levels of undesirable substances are compared.     

瓜叶绿素降解关键酶基因PPH和PAO cDNA片段的克隆与表达初步分析

 以西双版纳黄瓜和‘华黄5号’黄瓜叶片为试验材料,研究了衰老过程中两种黄瓜叶片叶绿素含量变化及脱镁叶绿素脱镁叶绿酸水解酶基因PPH和脱镁叶绿酸a加氧酶基因PAO的表达。结果表明：随着叶片衰老,叶绿素a和叶绿素b含量显著降低,叶绿素a与叶绿素b的比值在0.5左右,推测叶绿素b的含量可能对黄瓜叶片保绿起到重要作用。荧光定量RT-PCR分析结果显示：PPH和PAO在西双版纳黄瓜叶片生长35 d时表达量最高,分别为5.50和1.86;PPH在‘华黄5号’叶片35 d时表达量最高,为7.14,而PAO在35和45 d表达量都很高,分别为3.90和3.91;PPH和PAO在西双版纳黄瓜叶片中的表达量低于在‘华黄5号’中。     

Effects of simvastatin on PDGF-BB and serum-induced proliferation of vascular smooth muscle cells and on the expression of tumor suppressor gene PTEN

AIM:To observe the effect of simvastatin on the proliferation of vascular smooth muscle cells(VSMCs) induced by serum and growth factor PDGF-BB and the effect of simvastatin on the expression of PTEN,a important regulator of G₁/S cell cycle transition. METHODS:The DNA synthesis was determined by [³H]-TdR incorporation, cell cycle was examined with flow cytometry, the protein level of PTEN was measured by Western blot method. RESULTS: (1)Simvastatin inhibited [³H]-TdR incorporation in a dose dependent manner. (2) Flow cytometric DNA analysis revealed that simvastatin induced significantly enhancement of G₀/G₁ phase and decrease in S phase VSMCs.(3)Simvastatin increased protein level of PTEN and mevalonate, a metabolite of HMG-COA, reversed the effect of simvastatin on PTEN protein expression. CONCLUSION:Simvastatin may inhibit proliferation of VSMCs and retarded cell cycle in G₀/G₁ phase by increasing PTEN expression through inhibiting synthesis of mevalonate.     

Relationships of landscape,prey and agronomic variables to the abundance of generalist predators in cotton (<Emphasis Type="Italic">Gossypium hirsutum</Emphasis>) fields

A two-year field study investigated the possible effects of grain sorghum (Sorghum bicolor [L.] Moench) and uncultivated areas on the abundance of generalist predators in commercially-managed cotton (Gossypium hirsutum L.) fields in Texas, USA. From 63 to 70 fields were sampled for pests and predators over nine consecutive weeks during early stages of cotton development. Additional data on agronomic practices and landscape composition at three spatial scales were also collected for each field. Stepwise regression analyses were used to determine the relationships of landscape, agronomic and prey variables to the abundance of generalist predators. Because the variables most closely linked to predator levels could vary over time, separate regressions were conducted for three time periods corresponding to stages of grain sorghum growth (half-bloom, hard-dough, maturity) in each year. Significant relationships between predator abundance and agricultural landscape composition appear in both years and in all three time periods, but the specific relationships of landscape variables to cotton predator levels differed between and within years. At maturity in 2001, predator levels rose as the amount of uncultivated land from 1.6 to 3.2 km distant and the perimeter shared with grain sorghum increased. During 2002, the area of grain sorghum (half-bloom) and uncultivated land (hard-dough) within 1.6 km of cotton fields were both positively related to predator numbers. Cotton planting dates and the abundance of cotton fleahoppers (Pseudatomoscelis seriatus [Reuter]) were also strongly linked to predator numbers during both years. Results suggest that the total amount of grain sorghum or uncultivated land in an area is more important than the presence of these habitats adjacent to cotton fields, and that landscape composition may sometimes be the most important factor in determining predator abundance.     

Cryopreservation of shoot apices of hawthorn in vitro cultures originating from East Asia

The objective of this study was to establish a cryopreservation protocol for hawthorn shoot apices (Crataegus pinnatifida Bge.). Cryopreservation was carried out via encapsulation–dehydration, vitrification, and encapsulation–vitrification on shoot apices excised from in vitro cultures. We began by showing that cold-acclimation enhanced the regrowth of cryopreserved apices from 10.0 to 65.5% in encapsulation–dehydration. We then decided that the encapsulation–dehydration method was an optimal cryopreservation method for hawthorn shoot apices in terms of its high recovery after cryopreservation as well as its ease of use compared with vitrification and encapsulation–vitrification. In encapsulation–dehydration, the protocol leading to optimal regrowth was as follows: after cold-acclimation at 5 °C in the dark for 2 weeks, excised shoot tips were pretreated for 24 h at 25 °C on hormone-free Murashige and Skoog [Murashige, T., Skoog, F., 1962. A revised medium for rapid growth and bioassays with tobacco tissue culture. Physiol. Plant. 15, 473–497] (MS) basal medium with 0.4 mol/L sucrose, then encapsulated and precultured in liquid MS medium with 0.8 mol/L sucrose for 16 h at 25 °C. Precultured beads were dehydrated for 6 h at 25 °C in the dessicator containing 50 g silica gel to a moisture content of 15.3% (fresh-weight basis) before cryostorage for 1 h. In addition, we examined the effect of adding glycerol to both the alginate beads and loading solution to enhance regrowth after cryopreservation in encapsulation–dehydration. In the present study, it was shown that adding 0.5 mol/L glycerol resulted in high regrowth percentages (82.5–90.0%) in four Crataegus species.