白花蛇舌草总多糖的分离纯化、结构鉴定及初步免疫活性分析

目的:分离纯化白花蛇舌草总多糖,研究其对环磷酰胺致免疫低下小鼠的免疫增强活性。方法:采用超滤技术分离纯化白花蛇舌草总多糖,高效凝胶渗透色谱-示差检测器方法测定相对分子质量,柱前衍生化HPLC分析其单糖组成;采用环磷酰胺致免疫低下小鼠碳粒廓清试验研究其免疫活性。结果:从白花蛇舌草中分离得到1个单一多糖组分ODP-1,其相对分子质量20.88 k Da,单糖组成为甘露糖-鼠李糖-半乳糖醛酸-葡萄糖-半乳糖-阿拉伯糖,摩尔比0.005∶0.033∶0.575∶1.000∶0.144∶0.143;免疫活性试验结果表明,ODP-1可升高环磷酰胺致免疫低下小鼠廓清指数、吞噬指数、胸腺指数和脾指数。结论:白花蛇舌草多糖ODP-1为均一多糖,具有免疫增强活性。     

白及碱提多糖的提取分离及纯化研究

目的:探讨白及碱提多糖的分离纯化方法,研究其理化性质。方法:利用纤维素阴离子琼脂糖凝胶柱层析(DEAE-Sepharose Fast Flow)对白及多糖进行提取、分离和纯化,分别进行总糖量、糖醛酸含量和蛋白质含量测定,通过红外光谱、HPLC、GC-MS技术分析其成分、结构及理化性质。结果:从白及提纯得到5种分子量不同的白及多糖(BSPS)-Ⅰ～Ⅴ,分子量分别约为355.8、174.3、52.8、174.0、59.9 kDa;单糖组成摩尔比分别为BSPS-Ⅰ、Ⅲ:葡萄糖;BSPS-Ⅱ:甘露糖∶葡萄糖=1.05∶4.22;BSPS-Ⅳ:岩藻糖∶木糖∶甘露糖∶葡萄糖∶半乳糖=1.59∶0.70∶0.38∶1.22∶1.93;BSPS-Ⅴ:鼠李糖∶岩藻糖∶木糖∶甘露糖∶葡萄糖∶半乳糖=1.16∶1.93∶0.28∶0.21∶1.17∶1.50。结论:白及碱提多糖的单糖组成更加丰富。     

新月菱形藻胞外多糖的成分及其硫酸酯的制备

目的阐明新月菱形藻胞外多糖(exopolysaccharides of Nitzschia closterium,EPN)的成分、单糖组成及其硫酸酯化条件。方法新月菱形藻胞外多糖以醇沉法从该藻的培养液中制备;DEAE-Sephedex25柱分离,HPLC法测定其质量分数,HPLC测定其相对分子质量;以氯磺酸-吡啶法探索新月菱形藻硫酸酯化反应条件。结果新月菱形藻向胞外分泌3种多糖,相对分子质量分别为EPN11.886×105,EPN21.41×105,EPN31.134×105。3种多糖的单糖组成均为甘露糖、鼠李糖、葡萄糖醛酸、葡萄糖和岩藻糖。甘露糖、鼠李糖、葡萄糖醛酸、葡萄糖和岩藻糖的摩尔比在EPN1为4.3∶1∶5.1∶8.9∶5.6,EPN2为4.2∶7.3∶3.7∶12.8∶3.4,EPN3为3.5∶2.1∶4.3∶4.9∶4.7。3种多糖的硫酸酯化条件基本一致,提高氯磺酸比例和延长反应时间均有利于增加酯化产物的硫酸根、提高取代度和酸酯化反应的产率。结论新月菱形藻胞外多糖为酸性多糖,其硫酸酯化产物多糖硫酸酯可能具有潜在的药理活性。     

紫芝多糖GS-A-1的结构鉴定与生物活性研究

目的从紫芝液体深层发酵菌丝体精制多糖GS-A中进一步分离得到单一组分多糖GS-A-1,并研究其结构和生物活性。方法采用柱层析结合高效液相色谱分离,经HPGPC确定其分子量并证明该多糖组成的均一性。运用HPLC,高碘酸氧化和Smith降解,红外光谱及核磁共振光谱进行结构鉴定,同时通过荷瘤小鼠血常规检测初步评价其对环磷酰胺所致机体损伤的保护作用。结果 GS-A-1为单一组分的杂聚多糖,分子质量为21 190 Da,是一个由α-L-吡喃半乳糖和α-L-吡喃甘露糖通过(1→2),(1→6)连接而成,其摩尔比为半乳糖∶甘露糖=1.7∶1,且GS-A-1对环磷酰胺引起的化疗损伤具有保护作用。结论 GS-A-1为首次从紫芝液体深层发酵菌丝体的粗多糖中分离得到的在抗肿瘤药物中有应用价值的活性多糖。     

黄芪多糖分离与结构特征分析

目的从黄芪中提取分离多糖并对其进行理化性质及结构特征分析。方法采用DEAE-52和Sephadex G-100色谱柱分离纯化,得到黄芪均一多糖APS-Ⅰ和APS-Ⅱ,通过高效液相色谱(HPLC)、红外光谱(IR)和环境扫描电镜(ESEM)对APS-Ⅰ和APS-Ⅱ进行结构及形貌特征分析。结果 APS-Ⅰ和APS-Ⅱ为均一多糖,APS-Ⅰ中单糖组成为甘露糖、鼠李糖、葡萄糖醛酸、半乳糖醛酸、葡萄糖和半乳糖,且摩尔比率为29.12∶1.89∶4.00∶1.35∶1∶81.97;APS-Ⅱ中单糖组成为甘露糖、鼠李糖、葡萄糖醛酸、半乳糖醛酸、葡萄糖、半乳糖和木糖,且摩尔比率为50.46∶1.16∶1∶2.27∶2.66∶15.72∶7.86。IR显示APS-Ⅰ和APS-Ⅱ均为酸性多糖。ESEM观察显示APS-Ⅰ形貌工整,APS-Ⅱ形貌不规则。结论 APS-Ⅰ是一种呈丝带状结构分子量约为1.06×104Da的杂多糖,APS-Ⅱ是一种分子量约为2.47×106Da的杂多糖。     

柱前衍生HPLC法分析蔓菁多糖中单糖的组成

目的建立柱前衍生HPLC法测定藏药蔓菁多糖中单糖的组成。方法以2 mol/L硫酸水解蔓菁多糖,水解产物加入1-苯基-3-甲基-5-吡唑啉酮进行衍生化,采用反相高效液相色谱法,测定蔓菁多糖中单糖的衍生物。Kromasil C18色谱柱(250 mm×4.6 mm,5μm);乙腈为A流动相,0.1 mol/L磷酸盐(KH2PO4-Na OH,p H 6.8)缓冲液为B流动相,梯度洗脱;检测波长250 nm。结果蔓菁多糖由半乳糖、D-甘露糖、鼠李糖、阿拉伯糖、D-无水葡萄糖、D-葡萄糖醛酸、D-半乳糖醛酸7种单糖组成,其摩尔比为1∶0.26∶0.54∶1.06∶0.84∶0.05∶4.17。结论柱前衍生HPLC法表明蔓菁多糖主要由半乳糖醛酸、半乳糖和阿拉伯糖构成。     

柱前衍生HPLC分析黄连多糖的单糖组成

目的:建立柱前衍生HPLC测定黄连多糖中的单糖组成,并分析3种黄连多糖的单糖差异。方法:采用水提醇沉法提取黄连多糖,经硫酸水解后,用1-苯基-3-甲基-5-吡唑啉酮(PMP)柱前衍生,再利用HPLC法分析单糖的PMP衍生物。结果:黄连多糖主要由甘露糖、鼠李糖、葡萄糖醛酸、半乳糖醛酸、葡萄糖、半乳糖、阿拉伯糖7种单糖组成,其中半乳糖醛酸含量最高,葡萄糖次之。另外,雅连多糖、云连多糖和味连多糖中的7种单糖的平均摩尔比分别为1∶2.34∶0.38∶17.58∶12.40∶6.11∶5.93,1∶2.43∶0.25∶16.76∶15.18∶6.51∶9.78和1∶3.25∶0.40∶22.35∶12.96∶6.66∶10.28。结论:建立的方法简便、准确,重复性好,可用于黄连多糖中单糖的组成分析和含量测定。雅连多糖、云连多糖和味连多糖中的7种单糖含量有差异。     

柱前衍生化高效液相色谱法测定地稔多糖中单糖组成

目的:从地稔中提取分离地稔多糖,并对单糖组成进行研究。方法:采用水提醇沉法提取地稔粗多糖,再用DEAE纤维素柱色谱进行分离纯化,获得一种水溶性较好的中性多糖和一种酸性多糖,然后采用高效液相柱前衍生法对其单糖组成进行测定。结果:地稔中性多糖主要是由葡萄糖、半乳糖、甘露糖、阿拉伯糖、鼠李糖等5种单糖组成,摩尔比为123∶84∶27∶19∶5,另有极少量木糖。地稔酸性多糖单糖组成主要是半乳糖、半乳糖醛酸、葡萄糖、阿拉伯糖、甘露糖、葡萄糖醛酸、鼠李糖7种单糖组成,摩尔比为14∶10∶7∶6∶4∶1∶1,另外还有少量的木糖。结论:本研究建立了分离及测定地稔多糖组成成分的分析方法,该方法操作简单,方便可行。     

粗茎秦艽多糖GCP-1的分离纯化、结构表征及抗炎活性评价

目的从粗茎秦艽Gentianacrassicaulis中分离纯化多糖,初步分析其结构特征并研究其体外抗炎活性。方法采用热水浸提粗茎秦艽粗多糖(Gentianacrassicaulispolysaccharide,GCP),再经CelluloseDE-52纤维素离子交换柱、Sephadex G-100柱进行分离纯化,得到GCP-1。采用高效凝胶色谱法(HPGPC)鉴定纯度并测定其相对分子质量;采用高效液相色谱(HPLC)测定其单糖组成;采用红外光谱(IR)、气相色谱-质谱联用(GC-MS)和核磁共振波谱法对该多糖的结构进行初步分析;采用CCK-8法检测其对小鼠单核巨噬细胞RAW 264.7增殖的影响;利用ELISA试剂盒测定其对白细胞介素-1β(interleukin-1β,IL-1β)、一氧化碳(NO)和肿瘤坏死因子(tumor necrosis factor-α,TNF-α)释放量的影响。结果从粗茎秦艽中分离纯化得到GCP-1多糖,相对分子质量为6.87×10⁴,由甘露糖、鼠李糖、葡萄糖醛酸、半乳糖醛酸、葡萄糖、半乳糖和阿拉伯糖7种单糖组成,其物质的量比为0.07∶3.07∶0.42∶1.98∶1.33∶5.22∶6.76;IR、GC-MS和核磁共振波谱分析表明,GCP-1是一种典型的果胶多糖,含有HG和RGI区域及AGI和AGII侧链;质量浓度在10~200μg/m L时,GCP-1能明显促进小鼠单核巨噬细胞RAW 264.7的增殖(P<0.05),并能明显抑制小鼠单核巨噬细胞RAW 264.7分泌IL-1β、NO和TNF-α。结论GCP-1为从粗茎秦艽中分离得到的天然来源的均一多糖,其结构和活性初步研究为秦艽多糖成分的开发提供了一定参考。     

桑椹多糖的结构表征及其对乙醇脱氢酶活性的影响研究

该实验采用回流法提取桑椹多糖,经脱蛋白及DEAE-Cellulose 52分离纯化得到4种多糖组分(中性多糖MFP-1,微酸性多糖MFP-2,酸性多糖MFP-3和MFP-4)。通过化学法和仪器分析法对各多糖组分进行理化性质测定和结构表征,并研究其对乙醇脱氢酶(ADH)活性的影响。结果表明,MFP-1,MFP-2,MFP-3和MFP-4中的多糖量分别为75.3%,83.7%,79.1%,74.3%,其平均相对分子质量分别为112.2,128.8,199.5,181.9 k Da。MFP-1主要含有半乳糖、葡萄糖、阿拉伯糖、鼠李糖、甘露糖(26.8∶20.4∶8.7∶5∶1);MFP-2主要含有阿拉伯糖、半乳糖、鼠李糖、葡萄糖、半乳糖醛酸(34.2∶38.2∶8∶17.5∶15.1);MFP-3主要含有半乳糖醛酸、半乳糖、葡萄糖、鼠李糖、阿拉伯糖(28.3∶22.6∶20.9∶18.6∶12.5);MFP-4主要含有葡萄糖、半乳糖、鼠李糖、半乳糖醛酸、阿拉伯糖、葡萄糖醛酸(47.4∶34.9∶36.1∶33.1∶19.9∶4.1)。红外光谱分析结果表明,MFP-1,MFP-2,MFP-3和MFP-4中均有多糖的特征吸收峰,其中MFP-3和MFP-4是以β苷键为主的吡喃型多糖。活性结果表明,4种多糖对ADH均有激活作用,其ADH激活能力顺序为:联苯双酯MFP-3MFP-1MFP-2硫普罗宁MFP-4,MFP-1,MFP-2和MFP-3对ADH的激活作用均大于阳性对照硫普罗宁,同时发现MFP-3对ADH的激活效果与阳性对照联苯双酯的激活效果相当,推测其与之连接的糖醛酸种类有关,这为桑椹多糖的"构-效"研究提供参考依据。     

基于ICP-MS法的加味左金丸中无机元素分析

目的 通过电感耦合等离子体质谱（inductively coupled plasma mass spectrometry,ICP-MS）法建立加味左金丸中Cd、Pb、As、Hg、Co、V、Ni、Cu、Li、Sb、Ba、Mo、Sn、Cr、Na、Mg、Al、Ca、Ti、Mn、Fe、Zn、Ga、Se、Sr、Tl共计26种无机元素的测定方法。方法 加味左金丸通过微波消解法处理后,根据相对分子质量的大小选择内标物,其中⁷Li、²³Na、²⁴Mg、²⁷Al、⁴⁰Ca、⁴⁸Ti、⁵¹V、⁵²Cr、⁵⁵Mn、⁵⁶Fe、⁵⁸Ni、⁵⁹Co、⁶³Cu、⁶⁶Zn、⁷⁰Ga、⁷⁵As、⁷⁷Se、⁸⁶Sr以⁷²Ge作为内标;⁹⁵Mo、¹¹⁴Cd、¹¹⁸Sn、¹²¹Sb、¹³⁷Ba以¹¹⁵In作为内标;²⁰²Hg、²⁰⁵Tl、²⁰⁸Pb以²⁰⁹Bi作为内标。对标准品溶液、空白溶液与供试品溶液进行分析,采用标准曲线法进行定量分析。通过ICP-MS法进行测定。结果 26种无机元素线性的相关系数r ≥ 0.999 6,检出限为0.001~1.500μg/L,定量限为0.01~5.00 μg/L,精密度与重复性试验的RSD均小于5%,平均回收率在82.64%~106.44%,RSD均小于5%。对3个厂家的12批样品进行了测定,26种元素的含量差异较大,其中Na、Mg、Ca、Fe 4种元素的含量比较高,均大于500 μg/g,Cd、Pb、As、Hg、Co、Li、Sb、Mo、Sn、Cr、Se、Tl的含量比较低,均小于1 μg/g。由结果可知,人体的常量元素,如Na、Mg、Ca的含量比较高,Cd、Pb、As、Hg等有害元素含量比较低。根据《中国药典》2020年版一部的要求,本品中Cd、Pb、As、Hg与Cu均符合规定。结论 该方法快速、准确,可以用于加味左金丸中无机元素的测定。     

Erstaunliches Ergebnis einer Schädelakupunktur nach Yamamoto bei Hemihypsarrhythmie rechts

A girl, three and a half years old at the respective date, premature in 25th week of gestation, had been presented. On day three after birth, she had had a IVH III° right and left with severe involvement of parenchyma and other multiple complications, e. a. infant respiratory distress syndrome up to the 42nd day postpartum, and epileptic attacks with apnoeic events.     

七味红花殊胜散对缺血再灌注肝损伤的保护作用

目的::研究七味红花殊胜敬对缺血再灌注肝损伤的保护作用.方法:在大鼠肝缺血再灌注模型上,观察七味红花殊胜散对缺血再灌注肝损伤引起的血清丙二醛(MDA)、超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)、一氧化氮(NO)含量的变化.结果:七味红花殊胜散对大鼠血清MDA、SOD、GSH与再灌组比较有明显差异(P<0.05),NO含量与再灌组比较有明显差异(P<0.05).结论:七味红花殊胜散对大鼠急性缺血再灌注肝损伤具有保护作用.     

Methodology of Canine/Feline YNSA and Tail Acupuncture

BackgroundYamamoto New Scalp Acupuncture (YNSA) is a well-known acupuncture system for human patients which was developed by Dr. Yamamoto.ObjectiveMy aim was to transpose the human YNSA map onto canines and felines.MethodI started researching a canine/feline YNSA map in 2002. I investigated approximately 3,000 canines and felines for YNSA and 2,500 for Tail Acupuncture. The ratio of canines and felines was 6 to 4. I researched and found all the equivalent points by palpating animal patients on the basis of an exact Neck Diagnosis.ResultsI nearly completed the map in 2006. The points in the occipital region are incomplete and my research is still in progress. In the process of researching canine/feline YNSA points, I accidentally found the other microsystems around the tail and named it “Tail Acupuncture”.ConclusionsYNSA and Tail Acupuncture are acupuncture systems utilizing microsystems and support each other in canines and felines. The merits of YNSA and Tail Acupuncture are 1) immediate effects, such as in cases of lameness, 2) simplicity in finding the exact points to treat and 3) the scalp and the tail can be utilized separately or together as treatment zones, according to the situation.     

Chinesische Orthopädie: Teil 1: Die Zukunft der Akupunktur in der Postmoderne

The locomotor system is one of the major fields of application for acupuncture. The results of the large acupuncture studies funded by German Health Insurances from 2000 until 2006 are impressive and repressive: they restrict acupuncture application by allocation of reimbursement and create the necessity of reorientation, since acupuncture can neither be adequately described nor prescribed solely on the basis of disease entities. Regarding these disease entities, it seems rather necessary to analyse the layers of problems and formulate a hierarchy of therapeutic goals instead. Only in this way the effects of acupuncture, related techniques and further therapeutic modalities can be matched with the set of objectives. The article presents general principles and follows their application from biological aspects to implications for the locomotor system and consequences for the application of physical therapy. The manual of acupuncture focuses on therapeutic effects and objectives first, and then goes on to sketch simple therapeutic strategies and finally complex approaches within acupuncture. These may encompass acupuncture, microsystem acupuncture and complex therapeutic modes.     

Fallbericht: Patientin mit Schlafstörung

Background

Since the onset of her menopause about ten years ago, a sixty-year old patient has been suffering from hot flashes and sleep disorders causing her to become increasingly anxious and nervous. As the patient's mother had died of breast cancer, substitution of hormones was not advisable. Relaxation methods such as Yoga and Autogenic Training could only mildly alleviate the symptoms.

Objective

To effectively treat the sleep disorder, improve quality of life, lessen frequency and severity of hot flashes

Methods

Weekly sessions with ear- and body acupuncture for ten weeks

Results

After the third acupuncture session the patient reported a decrease of nightly waking episodes but suffered from acute pain in the neck as well as shoulder region. After nine therapy sessions, the patient reported a 50% decrease of nightly waking episodes. The hot flashes, however, remained unchanged.

Discussion

In this case, acupuncture significantly improved the patient's sleeping disorder. It also reduced the symptoms of anxiety and nervousness. Other menopausal symptoms such as hot flashes and dryness of mucous tissues, could not be influenced.     

Patientin mit chronisch rezidivierenden Gelenks- und Wirbelsäulenschmerzen

Background

Acupuncture as a regulative therapy is well-suited for the treatment of functional, reversible disorders. Additionally, the WHO recommends its application for pain therapy in cases of arthritis and back pain due to various causes. This case study should clarify which paths to take in the case of a multi-morbid patient suffering from chronic pains due to polyarthrosis.

Objective

Pain reduction and improvement of quality of life as well as improvement of sleep quality.

Methods

Annual treatment series consisting of 10 sessions each, one session per week.

Results

short-term pain reduction before undergoing endoprosthetic surgery; considerable improvement of general status and sleep.

Conclusions

Acupuncture may bring temporal relief in cases of chronic pain due to osteoarthritis and should therefore be applied in combination with western medicine. Acupuncture is particularly well-suited to treat the co-emergent vegetative symptoms.     

常见证候荷瘤小鼠神经内分泌免疫组织基因转录的总体特征

目的揭示常见证候荷瘤小鼠神经-内分泌-免疫组织基因转录的总体特征。方法采用小鼠标准化四诊及辨证方法，及Gene Chip Mouse Exon1．0ST Array等技术，检测H22荷瘤小鼠早期邪毒壅盛（邪毒）和气虚、中期阳气虚、中晚期气阴阳虚等4个常见证候下丘脑、垂体、肾上腺、睾丸、脾脏、胸腺、肿瘤RNA的转录与剪接。结果正常昆明种雄性小鼠下丘脑、垂体、肾上腺RNA电泳的28S峰低于18S，而睾丸、脾脏、胸腺、肿瘤相反。肿瘤发生的早期，下丘脑、垂体、肾上腺基因表达模式发生显著改变，邪毒尤甚，出现了失代偿。下丘脑28SRNA迅速抬升并持续，气虚强于邪毒。垂体RNA总量降低，28S陡降，邪毒尤甚。肾上腺RNA电泳特征类似垂体，气虚与邪毒相近，中晚期气阴阳虚尤甚。睾丸变化不明显。随着病情发展，脾脏重量持续增加，而胸腺相反，重量持续下降，蛋白合成和糖代谢下降。肿瘤组织蛋白合成与糖代谢，邪毒大于气虚。以上7个组织与正常对照组相比，有9127个基因表达发生差异、51126个外显子剪接发生差异。结论神经-内分泌-免疫网络组织基因转录的差异是荷瘤小鼠不同证候内在重要的物质基础。