用树脂法分离提取类芽孢杆菌(Paenibacillus)Cp-S316抗细菌活性物质

类芽孢杆菌Cp-S316对家蚕真菌、细菌病原物具有广谱抗性。为研究抗菌物质的化学结构、理化性质,并探讨其工业化生产的可行性,研究了不同树脂对抗细菌活性物质的吸附性能,对吸附性能优良的树脂进行了解吸性能的比较,并对解吸剂进行了筛选。结果:X-5树脂对活性物质具有较好的吸附性能,当树脂与发酵液混合后的质量浓度为0.03 g/mL时,振荡吸附4 h,其吸附率为90%;以0.02 mol/L HCl-75%丙酮为最佳解吸剂,层析柱柱长12 cm,直径1.5 cm,流速3.9 mL/(cm2.min)动态解吸,其总解吸率为92%,其中浓度较高部分(树脂与解吸液混合后的质量浓度为0.67 g/mL)的解吸率为90%。关键词类芽孢杆菌Cp-S316;活性物质;分离提取;树脂     

E12 technique: Conventional epoxy resin sheet plastination

Epoxy plastination techniques were developed to obtain thin transparent body slices with high anatomical detail. This is facilitated because the plastinated tissue is transparent and the topography of the anatomical structures well preserved. For this reason, thin epoxy slices are currently used for research purposes in both macroscopic and microscopic studies. The protocol for the conventional epoxy technique (E12) follows the main steps of plastination—specimen preparation, dehydration, impregnation and curing/casting. Preparation begins with selection of the specimen, followed by freezing and slicing. Either fresh or fixed (embalmed) tissue is suitable for epoxy plastination, while slice thickness is kept between 1.5 and 3 mm. Impregnation mixture is made of epoxy E12 resin plus E1 hardener (100 ppw; 28 ppw). This mixture is reactive and temperature sensitive, and for this reason, total impregnation time under vacuum at room laboratory temperature should not last for more than 20–24 hr. Casting of impregnated slices is done in either flat chambers or by the so‐called sandwich method in either fresh mixture or the one used for impregnation. Curing is completed at 40°C to allow a complete polymerization of the epoxy‐mixture. After curing, slices can be photographed, scanned or used for anatomical study under screen negatoscope, magnification glass or fluorescent microscope. Based on epoxy sheet plastination, many anatomical papers have recent observations of and/or clarification of anatomical concepts in different areas of medical expertice.     

A comparison of the efficiency of water and ethanol at removing formaldehyde from immersion fixed muscle tissues

The efficiency of water, ethanol solutions, isopropanol and ethylene glycol for extracting formaldehyde from fixed muscle tissue were compared. Similar samples of formalin preserved muscle were immersed in the test extraction fluids. After various storage periods the fluids were analyzed for formaldehyde content by sodium sulfite wet chemistry titration. The resulting data were compared by analysis of variance. Ethanol, isopropanol, ethylene glycol and water were found to be effective in extracting formaldehyde from formalin fixed tissue. The concentration of formaldehyde extraction by increasing concentrations of ethanol (20–40%) was linear. While 25 to 40% solutions of ethanol, 20% isopropanol and 25% ethylene glycol were significantly better at extracting formaldehyde, the actual percent formalin difference between water and the best extraction fluid (40% ethanol) was only 0.2% (2.43 versus 2.63%). This slight difference would hardly justify the added expense of using alcoholic or glycolic solutions solely for their extraction qualities. The problem of formaldehyde vapirs arising from embalmed tissue is associated with unbound and loosely bound formaldehyde. which can be removed by washing with various solutions. Precise methods for washing gross anatomical specimens are not discussed.     

粪便保存条件、时间对鸡球虫卵囊活力的影响实验

为探讨在不同温度条件下粪便保存不同时间对球虫卵囊成熟力的影响,本实验将含有球虫卵囊的粪便在-20℃、4℃、20℃、28℃分别保存0 h、24 h、48 h、72 h,通过观察卵囊的孢子化率确定最佳保存条件。结果表明:相同保存温度条件下,保存0 h、24 h、48 h、72 h的E.tenella早熟耐药株卵囊的孢子化率依次降低。-20℃和28℃时孢子化率均未达到80%;在4℃时粪便保存24 h的孢子化率达到80%以上,保存48h及以上时孢子化率极显著(P0.01)低于保存24 h的孢子化率;在20℃时的孢子化率均达到80%以上。     

Swine viscera preservation in hypersaturated salt solution after alcohol fixation as a preparation method for educational purposes

The use of live animals for educational purposes is an old practice that is still employed in teaching and research institutions. However, there are several objections to this practice, whether for ethical or humanitarian reasons. Surgical techniques teaching using anatomical pieces and/or preserved cadavers promotes greater learning efficiency, provides exercise repetition and increases the confidence and satisfaction of the students when compared to the use of live animals. The current work aimed to analyse the feasibility of using fresh swine urinary bladder and small intestines (jejunum), obtained from slaughterhouses, fixed in 99.8% ethyl alcohol (EA) and preserved in sodium chloride hypersaturated solution (SCHS) at 30%, for 7, 14 and 21 days, as an alternative method for surgical skills training (SST). Swine viscera, fixed in EA and preserved in SCHS, presented a realistic appearance, absence of odour and maintained the viable morphological characteristics during the performance of the operative techniques. Preservation solutions had low cost, were easy to acquire and did not offers risks to human health. Therefore, urinary bladders and small intestines fixed in 99.8% EA for 30 days and maintained in 30% SCHS at different periods were demonstrated as a good viable option as a preservation method for surgical skills training.     

茶叶总黄酮的提取工艺研究

黄酮类物质是茶叶的重要组成成分。本试验采用乙醇热提法提取茶叶中黄酮,结合正交试验设计研究时间、温度、乙醇浓度以及固液比4个因素对茶叶黄酮提取得率的影响。结果表明:各个因素对茶叶黄酮提取得率影响的主次顺序为乙醇浓度>固液比>温度>时间,茶叶黄酮的最佳提取条件为提取时间1h、提取温度80℃、乙醇浓度70%、固液比(g/ml)1:20。     

Effect of exposure duration of ovaries and oocytes at ambient temperature on parthenogenetic development of porcine follicular oocytes

This study was conducted to evaluate the effects of exposing porcine ovaries to 30-33 C during transportation for 4 h and subsequently room temperature (25 C) for 6 h of storage on in vitro maturation (IVM) and subsequent parthenogenetic development of oocytes collected from the ovaries. After IVM, oocytes having a tight oopalsm membrane and no signs of degeneration were exposed to Dulbecco's phosphate-buffered saline (DPBS) with 7% ethanol (v/v) for 7 min to induce parthenogenetic activation. Moreover, we also determined whether exposure of the collected oocytes to room temperature for 1, 2 and 4 h in DPBS or porcine follicular fluid (pFF) affected parthenogenetic development. When porcine ovaries were stored after transportation, oocytes collected from the stored ovaries showed a significantly higher rate of degeneration after 65 h of IVM (58.4%) and a significantly lower rate of cleavage after parthenogenetic activation (40.1%) than oocytes collected from ovaries immediately after transportation (38.9% and 47.4%, respectively). However, there was no significant difference in developmental rates to the morula and blastocyst stages between these two groups (14.4% and 14.3%, respectively). The duration of preservation, 1, 2, and 4 h, of oocytes in DPBS did not affect parthenogenetic development. In contrast, when preserved for 4 h in pFF, the developmental rates of the oocytes were significantly decreased. This suggested that some factor(s) in follicular fluid affects the developmental rate of oocytes with the passage of time in ambient conditions. These results suggest that even after 6 h storage of ovaries, oocytes having normal morphology after IVM have the same rate of parthenogenetic development as oocytes collected from ovaries just after 4 h of transportation, except for a lower cleavage rate, and that exposure of oocytes to room temperature for 4 h in DPBS does not affect their parthenogenetic developmental competence.     

冰川棘豆生物碱的提取分离

将冰川棘豆阴干 ,粉碎 ,过 2 0目筛。用 15倍量MeOH冷渗 ,减压回收溶剂 ,浓缩物加 0 1mol/LHCl溶解 ,过夜 ,过滤 ,酸水液以 80 0～ 10 0 0ml/ (min·cm2 )流经H+ 阳离子交换树脂。树脂出柱 ,水洗去悬浮物 ,上柱继续洗至无色 ,改用Me2 CO洗至流出液遇NH3 ·H2 O不变色后 ,将树脂倒入烧杯中 ,加 2mol/LNH3 ·H2 O适量 ,密闭 30min ,挥去NH3 ·H2 O ,再上柱 ,Me2 CO洗脱 ,回收溶剂得粗提物 ,粗提物用 1%HCl溶解 ,CHCl3 振摇洗涤数次 ,酸水层调pH 10 ,CHCl3 萃取 ,减压回收溶剂得粗碱 ,用石油醚溶出总碱 ,经硅胶G(含微量NaOH)柱层析 ,CHCl3 Me2 CO恒沸物洗脱 ,每 10ml收 1份 ,TLC检查 ,同类合并 ,石油醚重结晶 ,得晶Ⅰ和晶Ⅱ。并对它们的理化特性进行了初步测定。     

Detection of Ehrlichia risticii using an avidin-biotin-immunoperoxidase staining system

An indirect immunoperoxidase procedure using a specific anti-Ehrlichia risticii monoclonal antibody and an avidin-biotin-peroxidase staining method was used to detect E. risticii antigen in infected P388D1 murine monocytes. Several different methods of cytological fixation were used, including acetone (15 min), 95% ethanol (15 min), Bouin's fixative (5 hr), and 10% buffered neutral formalin (24 hr). The E. risticii organisms were labeled effectively and identified in cells fixed with acetone and ethanol. However, infected P388D1 cells fixed in 10% formalin or Bouin's fixative required enzymatic digestion with 1.0% trypsin for 15 min at 37 C before positive results were evident. This indirect immunoperoxidase avidin-biotin staining procedure proved to be a sensitive assay for the detection of intracellular E. risticii and may be an effective diagnostic procedure for formalin-fixed paraffin-embedded tissue.     

大孔吸附树脂分离纯化贯叶连翘中的金丝桃素

以贯叶连翘粗提浸膏为材料,考察六种大孔吸附树脂分离纯化金丝桃素的性能。采用静态、动态吸附方法筛选树脂,以中压分离方法确定分离纯化的条件。结果表明:HZ-801树脂以其高吸附率、高洗脱率成为优选的分离填料,其最佳分离纯化条件为:进样液质量浓度为0.1125 g/m L,进样速度为5.0 m L/min、洗脱速度为20 m L/min,0~95%乙醇溶液梯度洗脱,金丝桃素的纯度为79.11%。HZ-801可以较好分离纯化金丝桃素,纯化工艺具有较大实践性及参考价值。     

白三叶叶蛋白提取及纯化工艺

以白三叶（Trifolium repens）为材料探讨了叶蛋白的提取工艺和纯化方法。对白三叶叶蛋白提取中加热时间、温度、pH值、料液比、酸的种类等单因素以及纯化试剂甲醇、乙醇、丙酮、四氯化碳、水等进行了研究。结果表明,各单因素最佳参数为加热时间9 min,温度90 ℃,pH值 4.0,料液比1∶2,沉淀的酸为硝酸。对加热时间、加热温度、pH值和料液比进行4因素3水平正交试验,发现影响叶蛋白提取因素为pH值>温度>时间>料液比,最佳提取工艺为加热时间9 min,温度80 ℃,pH值4.0,料液比1∶2。对叶蛋白纯化结果表明,纯化剂对叶蛋白纯度的影响依次为甲醇>乙醇>丙酮>四氯化碳>水,但各种纯化剂之间差异不显著（P>0.05）。     

微波辅助提取山楂籽总黄酮的工艺研究

实验将微波辅助技术应用于山楂籽黄酮提取中,结果表明,浸提时间4.5 h、微波强度1 886.5 MHz、间歇作用7 min、水浴温度90℃、乙醇浓度60%、料液比1:20。该条件下,黄酮提取率可达89.5%,与传统回流提取法相比,黄酮提取率提高了20.2%。     

Dermal and serological response against Pseudomonas aeruginosa in sheep bred for resistance and susceptibility to fleece-rot

Genetically select lines of Merino sheep have been bred at Trangie (NSW Agriculture and Fisheries) for resistance (R) or susceptibility (S) to fleece-rot and flystrike. It is believed that fleece characters are primarily responsible for the R or S phenotype. When transferred to the wetter coastal environment of Sydney, R and S sheep with no more than 6 weeks wool cover, continued to show significant differences in the incidence and severity of fleece-rot dermatitis. To test the hypothesis that these sheep might also exhibit differences in their local skin reactions and immune responsiveness, 3 intradermal injections of killed Pseudomonas aeruginosa were administered at monthly intervals. After primary intradermal challenge, R sheep had a higher incidence of skin induration and a stronger inflammatory response (increased induration diameter) than S sheep. Compared to S sheep, R sheep also developed higher levels of circulating antibodies against whole cell antigen and both inner and outer membrane proteins of P. aeruginosa. These responses were maintained in R sheep with each consecutive challenge while S sheep showed a decline in their immune responsiveness. Differences in antibody response against outer membrane proteins were also detected when antigenically naive sheep from each genetic line were sensitised by epicutaneous challenge with P. aeruginosa under experimental wetting conditions. Intradermal challenge of these animals 6 months later with outer membrane proteins, revealed a late maximum (72 h) in the development of induration diameters for R sheep while S animals showed maximal induration diameters by 24 h. However, there was no significant difference in induration response between 24 h and 72 h within each group of sheep.(ABSTRACT TRUNCATED AT 250 WORDS)     

A comparison of fixatives suitable for scanning electron microscopy of Habronema spp

Members of the genus Habronema (Nematoda: Habronematidae) were preserved in 4 fixatives for examination with scanning electron microscopy (SEM). Tegumental features of the anterior and posterior extremities of these specimens were compared to evaluate the effect of the fixatives: modified Flemming's solution, glutaraldehyde (GA), Karnovsky's fixative and 70% ethanol. Fixatives were assessed on the appearance of the tegument, evidence of any wrinkling, shrinkage or swelling and the degree of extension in the male tail. Seventy per cent ethanol gave the most satisfactory results.     

粪便保存条件、时间对鸡球虫卯囊活力的影响实验

为探讨在不同温度条件下粪便保存不同时间对球虫卵囊成熟力的影响,本实验将含有球虫卵囊的粪便在．20℃、4℃、20℃、28℃分别保存0h、24h、48h、72h,通过观察卵囊的孢子化率确定最佳保存条件。结果表明：相同保存温度条件下,保存0h、24h、48h、72h的E．tenella早熟耐药株卵囊的孢子化率依次降低。-20℃和28℃时孢子化率均未达到80％;在4℃时粪便保存24h的孢子化率达到80％以上,保存48h及以上时孢子化率极显著（P〈0．01）低于保存24h的孢子化率;在20℃时的孢子化率均达到80％以上。     

木蝴蝶提取物制备及其抗菌抗炎活性的研究

本试验对木蝴蝶提取物的制备设计了6种提取方法,分别是水提取、微波提取、60%丙酮提取及95%、70%和45%乙醇提取,采用体外抑菌试验(药敏试验和最小抑菌浓度试验)来研究木蝴蝶的抗菌活性,进行了木蝴蝶急性毒性试验和最大耐受量试验,通过二甲苯致小鼠耳廓肿胀试验来观察木蝴蝶的抗炎活性。结果表明,木蝴蝶的6种提取物对金黄色葡萄球菌和鸡大肠杆菌都有抑菌效果,其中95%乙醇的提取物对金黄色葡萄球菌和鸡大肠杆菌呈高度敏感。木蝴蝶水提取物对小鼠安全无毒,其毒性小,最大耐受量为150 g/kg体重。木蝴蝶水提取物使小鼠耳肿胀度降低,表现出较好的抗炎作用,同时木蝴蝶水提取物使小鼠胸腺指数和脾脏指数降低,表明其对小鼠免疫功能有一定的抑制作用。     

The effect of region of application on absorption of ethanol and hexanol through canine skin

The effect of region of application on the percutaneous penetration of solutes with differing lipophilicity was investigated in canine skin. Skin from the thorax, neck, back, groin, and axilla regions was harvested from Greyhound dogs and placed in Franz-type diffusion cells. Radiolabelled (14C) ethanol (Log P 0.19) or hexanol (Log P 1.94) was applied to each skin section for a total of 5h. The permeability coefficient (kP, cm h(-1)) and residue of alcohol remaining in the skin were significantly (P=0.001) higher for hexanol compared to ethanol. In contrast, ethanol had a far greater maximum flux (Jmax, mol (cm2)(-1) h(-1)) than hexanol (P=0.001). A comparison of regional differences shows the kP and Jmax for ethanol in the groin was significantly lower (P=0.035) than the back. The kP and Jmax for hexanol were significantly higher (P=0.001) in the axilla than the other four skin sites. An understanding of factors influencing percutaneous drug movement is important when formulating topical preparations for the dog.     

菊科植物茼蒿有效成分提取工艺条件筛选

试验采用回流提取法,乙醇作为提取溶剂,按4因素3水平设计正交试验,筛选菊科植物茼蒿有效成分的最佳提取工艺条件。通过分析极差R值表明,乙醇浓度、萃取温度、萃取时间及固液比对得率影响的主次顺序是:乙醇浓度>萃取温度>固液比>萃取时间。最终确定菊科植物茼蒿有效成分最佳提取工艺条件是:乙醇浓度为55%,萃取温度为95℃,萃取时间为2h,固液比为1:8时得率最高。     

苍耳提取物对萝卜蚜和粘虫作用方式及解毒酶活性影响的研究

采用室内生测法,研究了苍耳3种溶剂(丙酮、乙醇、氯仿)提取物对萝卜蚜和粘虫的作用方式及苍耳氯仿提取物对两种试虫的3种解毒酶活性的影响。结果表明,苍耳氯仿提取物对萝卜蚜有很强的忌避作用,当浓度为0.05 g/mL,处理4 h,忌避率达94.25%,48 h忌避率为84.53%;其次是触杀作用,校正死亡率最高为67.24%,LC₅₀为0.7420 g/mL;内吸作用极弱。对粘虫主要表现为拒食和生长发育抑制作用,拒食校正死亡率最高达98.27%,AFC₅₀为0.1985 g/mL;72 h体重增长率为-5.85%,与对照差异极显著;胃毒和触杀毒力较弱,LC₅₀分别为0.8997和1.3070 g/mL。苍耳氯仿提取物对萝卜蚜和粘虫的乙酰胆碱酯酶(AchE)有抑制作用,处理萝卜蚜24 h及处理粘虫4 h时,乙酰胆碱酯酶(AchE)抑制率最高,分别为37.55%和25.65%;对谷胱甘肽转移酶(GSTs)和多功能氧化酶(MFO)活力影响不明显。     

Effects of Incubation Temperature and Semen Pooling on the Viability of Fresh,Chilled and Freeze‐Thawed Canine Semen Samples

This study assessed the effects of different incubation temperatures on semen viability and the influence of pooling on semen longevity. In experiment 1, semen samples were collected from five dogs, individually processed (individual semen: IS) and then aliquots from each male were pooled (pooled semen: PS). Semen samples (IS and PS) were diluted in a Tris‐glucose‐yolk extender and preserved as fresh (37 and 25°C) and chilled semen (4°C). Sperm motility and the percentages of sperm abnormalities and acrosome membrane integrity were assessed for 24 h. Storage at 25 or 4°C for the first 24 h yielded similar semen quality, but incubation at 37°C caused drastic reduction in sperm motility from 8 h of incubation onwards. In experiment 2, the semen was processed in the same way to that of experiment 1 and then preserved at 25 or 4°C until semen inactivation. Semen that was incubated at 25°C became completely inactive after 3–4 days of storage, while semen that was preserved at 4°C presented with more gradually decreased sperm motility (mean values of 40–60% for the first 8 days). In addition, the mixing of semen was only observed to influence the sperm quality of the samples stored at 4°C. In experiment 3, semen was collected from five dogs, pooled and frozen in liquid nitrogen; after thawing, it was preserved at 37, 25, 15 and 4°C, and the sperm quality was defined. The motility of the freeze‐thawed semen samples decreased quickly in the first 4 h after thawing, regardless of the preservation temperature of the thawed semen. This study confirmed that semen preserved at 37°C should be used within a maximum of 12 h, while the semen stored at 25°C shows acceptable quality for 24 h. Chilled semen presented highest most sustainable quality, especially when semen is processed as pooled semen.