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。     

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.     

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.     

Ergebnisse der Leistungsprüfungen der Süßkirschensorte ‚Stella‘ auf Gisela- und Weiroot-Unterlagen in Bulgarien

Zusammenfassung Die Leistungsprüfungen wurden im Zeitraum 1997 bis 2003 mit den Unterlagen Gisela 4 und 5, den Klonnummern 195/20 und 497/8 aus der Gisela-Serie sowie Weiroot 10, 13, 53, 72 und 158 durchgeführt. Dabei dienten Sämlinge von P1 (bulgarische Selektion aus Prunus mahaleb) als Kontrolle. Alle Unterlagen waren mit der Sorte Stella veredelt und im Dezember 1996 in der Versuchsanlage der Agraruniversität in Plovdiv, Bulgarien, im Abstand von 6 m×4,5 m gepflanzt worden. Dabei erfolgte ein Pflanzschnitt. Nach Abschluss der natürlichen Kronenentwicklung wurde jedes Jahr ein Winterschnitt vorgenommen. Der Boden wurde durch mechanische Bearbeitung offen gehalten und nach dem 4. Standjahr wurden die Baumstreifen mit Herbiziden behandelt. Die Wasserversorgung erfolgte durch eine dem natürlichen Gefälle folgende Überflutung, allerdings nicht immer zum optimalen Zeitpunkt, da keine eigene Wasserquelle zur Verfügung stand.Basierend auf den Ergebnissen bis zum Anfang des 7. Standjahres können die untersuchten Unterlagen in zwei Gruppen differenziert werden: starkwüchsig—Weiroot 10, P1 und Weiroot 13; mittelstarkwachsend bis schwachwüchsig—Gi 497/8, Gisela 4, Weiroot 53, Weiroot 158, Gi 195/20, Weiroot 72 und Gisela 5. Letztere zeichnete sich durch besondere Schwachwüchsigkeit aus. Die meisten Wurzelschosser bildeten Gisela 4, Weiroot 10 und Weiroot 13. Weiroot 53, Weiroot 72 und Weiroot 158 entwickelten deutlich weniger und P1, Gisela 5, Gi 195/20 sowie Gi 497/8 keine Wurzelschosser. Den frühesten Blühbeginn induzierte Gisela 4. Die anderen Unterlagen führten, in Abhängigkeit von den Temperaturbedingungen des jeweiligen Jahres, zu einer Verspätung der Blüte: P1 und Weiroot 10 um 1–2 Tage; Gi 497/8, Weiroot 13 und Weiroot 158 um 2–4 Tage; Weiroot 72 um 2–7 Tage; Gi 195/20 um 3–6 Tage; Weiroot 53 um 3–8 Tage und Gisela 5 um 3–10 Tage. Die Reifezeit der Früchte war bei den Bäumen auf Gisela 5 im Vergleich zu den anderen Varianten um 2–3 Tage verspätet. Gisela 5, Weiroot 72 und Gisela 4 induzierten bei der aufveredelten Sorte die höchsten Ertragsleistungen, P1 die geringsten. Bei den Bäumen auf Gisela 5 war die Fruchtgröße geringer als bei den anderen Unterlagen. Bäume auf Gisela 5 brauchen intensive Pflege. Nur wenn alle Produktionsfaktoren und kulturtechnischen Maßnahmen optimiert werden, kann das hohe Ertragspotenzial dieser Unterlage ausgeschöpft werden.     

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.     

多效唑对猕猴桃离体试管苗生长及内源激素的影响

多效唑（ＰＰ３３３）处理猕猴桃试管苗，降低了其生长强度；植株体内的ＧＡ３、ＩＡＡ和ＺＴ含量下降，ＡＢＡ的含量上升，乙烯释放率增加；并且能降低外源的ＧＡ３和ＩＡＡ促进生长的作用，而外源的ＧＡ３和ＩＡＡ又能不同程度地逆转多效唑的抑制作用，使植株恢复生长。     

Proteomic analysis between pathological scars and normal skin

AIM: To investigate and screen the sensitive proteins in the formation mechanism of pathological scars by comparing the results of differential proteomic analysis between pathological scars and normal skin.METHODS: Two-dimensional gel electrophoresis was used to detect the protein expression profiles in 8 keloid patients, 8 hypertrophic scar patients and 3 matched normal skin patients.The proteins that showed differential expression of over 4-fold change were cut and analyzed by MALDI-TOF/TOF mass spectrometry.RESULTS: A two-dimensional protein profiling comparison between pathological scars and normal skin was successfully established.On average, 2 978 spots in keloid, 2 975 spots in hypertrophic scar and 3 053 spots in normal skin were identified using gel analysis software.Compared with normal skin, there were totally 36 differentially-expressed proteins in keloid and hypertrophic scar identified from the spots of over 4-fold change, including 16 proteins in both keloid and hypertrophic scar (8 up-regulated and 8 down-regulated), 11 only in keloid (9 up-regulated and 2 down-regulated) and 9 only in hypertrophic scar (4 up-regulated and 5 down-regulated).CONCLUSION: Proteomic analysis can identify the proteins with variance of pathological scars versus normal skin, thus providing probable new clues to reveal the formation mechanism of pathological scars.     

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.     

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.     

Coupling past management practice and historic landscape change on John F. Kennedy Space Center,Florida

Historic landcover dynamics in a scrubby flatwoods (Tel-4) and scrub landscape (Happy Creek) on John F. Kennedy Space Center were measured using aerial images from 1943, 1951, 1958, 1969, 1979, and 1989. Landcover categories were mapped, digitized, geometrically registered, and overlaid in ARC/INFO. Both study sites have been influenced by various land use histories, including periods of range management, fire suppression, and fire management. Several analyses were performed to help understand the effects of past land management on the amount and spatial distribution of landcover within the study sites. A chi-squared analysis showed a significant difference between the frequency of landcover occurrence and management period. Markov chain models were used to project observed changes over a 100-year period; these showed current management practices being effective at Tel-4 (restoring historic landscape structure) and much less effective at Happy Creek. Documenting impacts of past management regimes on landcover has provided important insight into current landscape composition and will provide the basis for improving land management on Kennedy Space Center and elsewhere.     

XBP-01 accelerated apoptosis induced by oxidized low density lipoprotein in vascular smooth muscle cells

AIM: Previous studies performed with XBP-01 in vitro indicated that XBP-01 could inhibit vascular smooth muscle cells from being transformed into foam cell and could eliminate the atherosclerotic plaque in C57BL/6J mouse. This experiment is to investigate its mechanism of eliminating plaques in vitro. METHODS: The cultured porcine artery smooth muscle cells incubated with XBP-01 of 0.1 mg/L for 24 h after preincubated with oxidized low density lipoprotein of 15 mg/L for 72 h in vitro. The samples were analyzed by fluorescence microscope, confocal microscope system and flow cytometry. RESULTS: Apoptosis was triggered by being incubated with oxidized low density lipoprotein and this process was accelerated additionally by being incubated with XBP-01. CONCLUSION: XBP-01 can be effective in eliminating atherosclerotic plaque by accelerating the process in which oxidized low density lipoprotein induced smooth muscle cell apoptosis.