Simultaneous Detection of ACP1 and GC Genotypes Using PCR/SSCP

The classical enzyme and protein markers ACP1 and GC have gained new importance because of the biological functions of their gene products. ACP1 encodes a low molecular weight enzyme which is now recognized as a phosphotyrosine phosphatase with a role in the regulation of signal transduction pathways, and GC‐globulin acts both as a transporter of vitamin D and as a plasma actin scavenger and plays a role in macrophage activation. These two polymorphisms were phenotyped for decades on the basis of electrophoretic isozyme or protein patterns; the gene structures are now known. Nucleotide substitutions determining the common alleles are close enough at each locus to be contained in one short PCR product. We have developed a simple, rapid and reliable multiplex method based on PCR and SSCP which allows the simultaneous determination of the common ACP1 and GC genotypes.     

Expression of classical protein kinase C subspecies in non-neoplastic lymphocytes and non-Hodgkin's lymphomas: an immunohistochemical study

It is generally accepted that phosphorylation plays a pivotal role in the cellular response of cell differentiation and proliferation. Immunohistochemical expression of classical protein kinase C (cPKC) subspecies (alpha, beta and gamma) in eight reactive lymphoid tissues, three normal spleens and 149 non-Hodgkin's lymphomas was examined. cPKC beta was observed primarily in the mantle zone B cells, but appeared as very faint staining in Ki-67 positive proliferated B cells in the germinal centers of secondary lymph follicles. In contrast to the reactive state, high levels of cPKC subspecies were recognized in the majority of 149 cases of non-Hodgkin's lymphoma, including those thought to have arisen from germinal center cells such as follicular lymphoma. The expression of cPKC alpha was found in higher frequency in T cell lymphomas than B cell lymphomas (P < 0.01) by the Chi-squared test. High levels of cPKC alpha were present only in high grade or highly aggressive lymphomas, showing the highest incidence in the small non-cleaved cell type, according to the International Working Formulation and National Cancer Institute (P < 0.01). cPKC gamma was not detected in normal lymphoid cells and was expressed in only four cases of non-Hodgkin's lymphomas. It is presumed that cPKC alpha and beta have a relationship to cell activation and proliferation of lymphoid cells of reactive and neoplastic states. It might be considered that the expression of cPKC alpha may have a relationship with aggressiveness in non-Hodgkin's lymphomas.     

Early coping experience and later aversive conditioning in cats

Kittens were given differential early experience in order to compare an objective coping behavior with the result of an inescapable aversive experience. Separate groups of kittens were treated in a shock motivated runway task at either 4 or 12 weeks of age, by allowing one member of a weight matched sibling pair to acquire an escape behavior, while the other member was confined; a third subject served as a handled control. Escape behavior was significantly different for 4 and 12 week old subjects, since the older kittens reached a running asymptote within the first few shock trials. At 6 months of age, the subjects were tested for effects of differential early treatment; heart rate, respiration rate and amplitude, and somatic activity were measured during classical conditioning. While all groups gave evidence of acquisition in one or more response measures, only a potentiated heart rate response in 4 week kittens could be related to early experience. Heart rate did not differentiate escaping kittens from confined ones. Rather, heart rate was related to early treatment with shock, perhaps reflecting an increased tendency to react with a passive defensive response.     

经典名方中藿香类药材的本草考证

通过查阅古代相关本草、医籍、经史及近现代文献资料,笔者对藿香类药材的名称、基原、产地、品质评价、栽培、采收加工及炮制方法等进行了系统梳理与考证,可为含藿香类药材的经典名方开发利用提供依据。通过本草考证分析可知,历代本草多以“藿香”为正名,所用主流基原均为Pogostemon cablin,为区别于明代以来我国民间习用的同科另一种植物土藿香Agastache rugosa及推崇道地产区而有“广藿香”之名;广藿香原产于越南等东南亚国家,早期因作香料经广东等地传入我国,宋代以来便已在我国南方引种成功;药用部位多为其干燥地上部分,亦有将其叶、梗分开单独使用;历代推崇的道地产区为广东,现广东广州、肇庆、湛江和海南省多有栽培,以石牌所产最为知名;广藿香主要为扦插繁殖,二月生苗,六月采收;产地加工方法主要为闷晒法,放置阳光下反复闷晒至有芳香气味、颜色转黄;炮制方法多以净选后生品入药。基于考证结果,建议养胃汤中所用藿香以P. cablin为基原,取叶片生品入药,而藿朴夏苓汤则建议选择土藿香A. rugosa除去根及老茎的生品入药。     

基于古今文献对经典名方中细辛毒性的有效避减探析

目的：通过研究细辛的相关文献,对影响细辛毒性的关键信息进行梳理,为经典名方中细辛的合理应用提供依据。方法：通过文献计量学的方式,搜集细辛的古今相关文献,分析细辛毒性与药物基原、用药部位、炮制方式、药物剂型、方药配伍、服药方法及患者体质因素之间的关系。结果：在经典名方的研发过程中,细辛在当归四逆汤、厚朴麻黄汤中的用量分别为9、6 g,煎煮时间应>120 min;细辛在辛夷散、三痹汤、大秦艽汤、清上蠲痛汤种的单次用量分别为0.8、1.2、0.9、1.1 g;当归四逆汤、厚朴麻黄汤、清上蠲痛汤等应选用辽细辛的根茎,辛夷散可选择汉城细辛的根茎。在药物的炮制上,当归四逆汤、厚朴麻黄汤、三痹汤、大秦艽汤四方中的细辛可选用酒制;辛夷散、清上蠲痛汤中的细辛可选用炒制;另外,细辛的毒性与药物的配伍和患者的体质等因素均有着密切的关系。结论：该研究通过梳理有关细辛毒性的文献资料,得出了影响细辛毒性的关键信息,探析了细辛毒性的有效避减方式,为包含细辛的经典名方的合理开发和安全应用提供了更为充分的依据。     

经典名方中海风藤的本草考证

通过查阅古代本草、医籍,结合近现代文献资料,笔者对海风藤药材名称、基原、产地及采收加工进行考证,以期为含海风藤药材的经典名方开发提供依据。经考证表明,海风藤入药最早以“南藤”为正名收载于唐代《本草拾遗》,并有“丁公藤”“石南藤”等异名;而“海风藤”一名应出现于明代;宋代之前海风藤药材来源可能为胡椒科胡椒属植物石南藤Piper wallichii的藤茎,宋代后使用品种则多为胡椒属风藤P. kadsura、山蒟P. hancei等植物,历版《中华人民共和国药典》收载海风藤药材基原仅风藤P. kadsura一种,结合基原考证、市场调查及野生资源分布情况,建议经典名方所用海风藤基原除风藤P. kadsura外,可增加山蒟P. hancei;由于气候变化及胡椒属植物喜热的习性特点,历代海风藤药材所著录的产地呈现由秦岭一带逐渐向南迁移至南方盛产胡椒属植物的地区,明代以来则推崇福建省泉州地区;采收期为农历七月至八月,采取地上部分,除去须根、细茎及叶,晒干;历代关于海风藤药材的炮制记载较少,多为生品入药,故建议经典名方所用海风藤药材未注明特殊炮制要求的采用生品即可。     

基于经方解构新说“石膏配伍解肌法”

石膏解肌之说由来已久,但限于矿物药的溶出及研究手段有限等现实问题,其临床推广及应用不足,深入探讨亦不足。该文通过梳理解肌法的脉络,总结出解肌法所针对的病机为“津液不足,兼表不和”,而石膏解肌的核心内涵为“清火存液,兼透表邪”;通过梳理解肌法的古今运用,结合配位化学的现代研究,从配伍角度入手,主以《伤寒论》和《金匮要略》中的经方为例,拓展了石膏解肌之内涵,总结出石膏的解肌作用不仅可以治疗外感后的肌肉不适症状,还可能影响脏腑功能,进而治疗内伤杂病,并由此提出“石膏配伍解肌法”,认为石膏与麻黄、葛根、防己、知母、人参、芍药、竹茹、生地黄、麦冬和葳蕤等药物配伍后,通过调整透散和清热的力度来更好地发挥解肌作用。该文附临床验案一则予以阐发,以期为石膏的临床运用提供参考,为其配伍使用的药物机制研究提供方向。     

基于数据挖掘探讨全国名中医王行宽诊疗胸痹心痛遣方用药规律及学术经验

目的：运用中医传承计算平台V3.0软件,研究王行宽教授治疗胸痹心痛遣方用药规律,传承王行宽教授诊疗胸痹心痛的学术经验。方法：收集整理2017—2020年王行宽教授于湖南中医药大学门诊诊疗冠心病心绞痛患者的原始病历资料,录入中医传承计算平台V3.0,运用软件进行方药规律研究。结果：共收集王行宽教授治疗胸痹心痛处方1 044则,所用药物多为甘、苦药物,归经以肺经为主,其次为心、脾、肝、胃、肾经;所用方剂中使用最多的经方是生脉散,最高的经验方是心痛灵Ⅲ号方;高频数药物主要有麦冬、半夏、丹参、瓜蒌皮、黄连、五味子、柴胡等药物;药物的常用剂量多为3、5、10、15 g;组方规律分析得到常用药组合129个,置信度>0.99的组合有58个,并得到常见证型核心药物;药物聚类得到6个核心药物组合。结论：王行宽教授论治胸痹心痛以益气养营、豁痰化瘀、疏肝利胆为治疗思路,并根据胸痹心痛不同证型予以辨证施治,体现其“多脏调燮、综合治理”的学术思想,其核心处方可供临床从业者参考,但仍需要进一步的临床及实验研究验证其疗效。     

经典名方中石决明的本草考证

通过查阅历代本草、医籍、方书及现代文献资料,笔者对经典名方中所用石决明药材进行系统本草考证,涉及名称、基原、产地、品质评价、采收加工及炮制等方面内容,以期为含石决明的经典名方开发提供依据。经考证可知,历代皆以石决明为正名,亦有以其功效、性状及近音字而命名的“千里光”“九孔螺”“珍珠母”等异名。唐代以前石决明药材动物基原因形态描述过于简练,仅能考订为鲍科鲍属动物;宋元明清时期主流基原为杂色鲍Haliotis diversicolor与皱纹盘鲍H. discus hannai;民国时期开始将鲍属多种动物作为石决明基原使用,品种繁杂并延续至今。古今我国国内主要产区为海南、广东及山东地区,国外产地主要为日本及越南。古籍中对石决明品质评价大致以呼水孔的开孔数量为准,以七孔和九孔的鲍壳为佳;近现代则以性状个大整齐、无破碎、内外洁净并有光彩、壳厚者为佳。基于考证结果,结合石决明散的记载年代,建议开发该经典名方时石决明药材使用杂色鲍H. diversicolor或皱纹盘鲍H. discus hannai的贝壳,且以生品入药。     

经典名方中杜仲的本草考证

通过查阅历代本草、医籍、方书并结合近现代文献资料,笔者对杜仲药材的名称、基原、产地、品质评价、采收加工、炮制历史沿革及产地变迁情况进行了系统梳理及考证,以期为含杜仲的经典名方开发提供参考依据。经考证可知,历代本草均以杜仲为正名,基原为杜仲科植物杜仲Eucommia ulmoides的干燥树皮,古今一致;杜仲最早的产地为河南、山西、陕西、四川一带,自明代以来产地扩展至全国大部分地区,且推崇四川、陕西、重庆、贵州、湖北等地为杜仲的道地产区;近代以来总结其品质以皮厚、块大、粗皮刮净、断面多丝、内表面色暗紫为佳;杜仲的古代炮制加工方法主要有去粗皮切制生用和加酥蜜、姜汁、盐水、酒等辅料炮制,近现代以来炮制方法日趋简化,沿用的炮制方法主要为净制后切制生用、盐炙,建议挖掘不同杜仲炮制品的现代科学内涵,通过标准恢复传统主流炮制方法。基于宋代陈自明的三痹汤中杜仲“去皮,切,姜汁炒”的要求,根据考证结果,建议使用姜杜仲,即参考2020年版《中华人民共和国药典》炒法,以姜汁为辅料进行炮制后入药。