目的 以活性成分为指标,优化滇白珠抗炎镇痛活性部位(ARF)提取富集工艺.方法 以水杨酸甲酯三糖苷(MSTG-B)为指示性成分对滇白珠水杨酸甲酯糖苷类成分进行提取及富集,确定最佳提取和洗脱溶剂.选用Thermo Hypersil Gold C18(250 mm×4.6 mm,5μm)色谱柱,以乙腈-0.2％乙酸溶液为流动相进行梯度洗脱,流速为1.0 ml/min,检测波长为294 nm,建立滇白珠ARF部位的HPLC指纹图谱.结果 滇白珠药材的最佳提取溶剂为30％乙醇溶液,经大孔树脂AB-8富集后的35％乙醇洗脱部位为ARF部位.所测定的3个不同产地的滇白珠样品,其ARF部位的指纹图谱相似度和方法学考察均符合要求.结论 ARF部位提取富集方法工艺稳定,重现性好.

目的:建立高效液相色谱法(HPLC)测定民族药滇白珠抗炎镇痛活性部位(ARF)中4种活性成分methylbenzoate-2-O-β-D-xylopyranosyl(1-2)[O-β-D-xylopyranosyl(1-6)]-O-β-D-glucopyranoside(MSTG-A),methyl benzoate-2-O-β-D-glucopyranosyl(1-2) [O-β-D-xylopyranosyl(1-6)]-O-β-D-glucopyranoside(MSTG-B),gaultherin及绿原酸的含量测定方法,为滇白珠药材的深入研究及其质量控制提供方法依据,并为后期滇白珠的制剂与临床应用奠定基础.方法:采用COSMOSIL 5C18-PAQ色谱柱(4.6 mm×250 mm,5μm),以甲醇-0.2％冰乙酸水溶液为流动相梯度洗脱,柱温25℃,流速l mL· min-1,检测波长294 nm.结果:MSTG-A,MSTG-B,gaultherin,绿原酸分别在0.014 06 ～0.450 00,0.007 81 ～0.250 00,0.003 13 ～0.100 00,0.000 94～0.030 00 g·L-1线性关系良好(r≥0.999 7);方法精密度、重复性、稳定性良好;平均加样回收率分别在100.81％,99.19％,96.12％,102.56％,其RSD分别为1.4％,0.7％,0.7％,2.4％.滇白珠ARF部位中MSTG-B,MSTG-A,gaultherin,绿原酸质量分数分别为23.608,41.973,8.282,2.673 mg·g-1.结论:该方法操作简便、准确、重复性好,适用于滇白珠抗炎镇痛活性部位中4种活性成分的含量测定,为滇白珠药材的深入研究、质量控制与综合评价提供参考依据,并为后期滇白珠药材的制剂与临床应用奠定基础.

目的 考察滇白珠抗炎镇痛活性部位(ARF)的肠吸收特性.方法 采用大鼠右后足足底注射完全弗氏佐剂方法建立佐剂诱导关节炎(AIA)动物模型,造模成功后将模型大鼠随机分为3组(ARF高、中、低浓度),每组3只,另取9只正常大鼠随机分为3组(ARF高、中、低浓度),每组3只;实验前12 h禁食不禁水,收集大鼠肠系的小肠部分10 cm左右,采用大鼠离体外翻肠囊法,考察滇白珠ARF高、中、低浓度给药后正常大鼠和AIA模型大鼠不同时间点的肠吸收差异.通过HPLC检测肠囊液样品中3个水杨酸甲酯糖苷类成分methyl benzoate-2-O-β-D-xylopyranosyl(1-2)[O-β-D-xylopyranosyl(1-6)]-O-β-D-glucopyranoside(MSTG-A)、methyl benzoate-2-O-β-D-glucopyr-anosyl(1-2)[O-β-D-xylopyranosyl(1-6)]-O-β-D-glucopyranoside(MSTG-B)及2-[(6-O-β-D-xy-lopyranosyl-β-D-glucopyranosyl)oxy]benzoic acid methyl ester(冬绿苷)的含量,计算累积吸收量,吸收速率常数及表观渗透系数,比较3个成分在正常大鼠及AIA模型大鼠的肠吸收情况.结果 滇白珠ARF中的3个活性水杨酸甲酯糖苷类成分在正常大鼠及AIA模型大鼠中均为线性吸收(均r>0.9),符合零级吸收速率.各成分吸收速率常数与给药浓度及机体状态有关,在生理状态下,其吸收速率常数均随ARF浓度的增加而增加,说明3个指标成分属于被动扩散;而在病理状态下,其吸收速率常数均随浓度的增加先减小后增加.各成分在生理及病理状态下均吸收良好,表观渗透系数均大于1×10-6 cm/s,且在病理状态下吸收强于生理状态.结论 滇白珠ARF中MSTG-A,MSTG-B和冬绿苷在正常及AIA模型大鼠离体小肠中的吸收均符合零级动力学过程,且病理状态对活性成分的吸收具有影响;选择外翻肠囊法可评价滇白珠ARF中3个水杨酸甲酯糖苷类成分的肠吸收特性.

Multiple plant lineages have independently evolved sex chromosomes and variable kary-otypes to maintain their sessile lifestyles through constant biological innovation.Morus notabilis,a dioecious mulberry species,has the fewest chromosomes among Morus spp.,but the genetic basis of sex determination and karyotype evolution in this species has not been identified.In this study,three high-quality genome assemblies were generated for Morus spp.[including dioecious M.notabilis(male and female)and Morus yunnanensis(female)]with genome sizes of 301-329 Mb and were grouped into six pseudochromosomes.Using a combination of genomic approaches,we found that the putative ancestral karyotype of Morus species was close to 14 protochromosomes,and that sev-eral chromosome fusion events resulted in descending dysploidy(2n=2x=12).We also charac-terized a～6.2-Mb sex-determining region on chromosome 3.Four potential male-specific genes,a partially duplicated DNA helicase gene(named MSDH)and three Ty3_Gypsy long terminal repeat retrotransposons(named MSTG1/2/3),were identified in the Y-linked area and considered to be strong candidate genes for sex determination or differentiation.Population genomic analysis showed that Guangdong accessions in China were genetically similar to Japanese accessions of mul-berry.In addition,genomic areas containing selective sweeps that distinguish domesticated mul-berry from wild populations in terms of flowering and disease resistance were identified.Our study provides an important genetic resource for sex identification research and molecular breeding in mulberry.