1.Comparison of Wild and Cultivated Bupleurum scorzonerifolium Based on Traditional Quality Evaluation
Changsheng YUAN ; Feng ZHOU ; Xingyu LIU ; Yu SHI ; Yihan WANG ; Huaizhu LI ; Yongliang LI ; Shan GUAN ; Huaizhong GAO ; Yanmeng LIU ; Zhilai ZHAN
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(5):203-214
ObjectiveTo characterize the quality differences among different germplasm and introduced varieties of Bupleurum scorzonerifolium roots(BSR), and explore the underlying molecular mechanisms, providing a basis for high-quality production and quality control. MethodsWild BSR from Yulin(YLW) served as the quality reference, we conducted comparative analysis among YLW, locally domesticated wild germplasm in Yulin(YLC3), Daqing germplasm introduced and cultivated in Yulin(YLDQC3), and locally cultivated germplasm in Daqing(DQC3). A combination of traditional pharmacognostic methods and modern multi-omics analyses was employed, including macroscopic traits(appearance, odor), microscopic features(proportions of cork, phloem, xylem), cell wall component contents(hemicellulose, cellulose, lignin), carbohydrate contents(starch, water-soluble polysaccharides), marker compound contents(ethanol-soluble extracts, total saponins, liposoluble extracts, and saikosaponins A, B2, C, D), metabolomics, and transcriptomics, in order to systematically characterize quality differences and investigate molecular mechanisms among these samples. ResultsMacroscopically, Yulin-produced BSR(YLW, YLC3, YLDQC3) exhibited significantly greater weight, length, and upper and middle diameters than Daqing-produced BSR(DQC3). Odor-wise, YLW and YLC3 had a a fragrance taste, YLDQC3 had a rancid oil odor, and DQC3 had a sweet and fragrant taste. Microscopically, Yulin germplasm(YLW, YLC3) and Daqing germplasm(YLDQC3, DQC3) shared similar structural features, respectively. However, Yulin germplasm showed significantly higher proportions of cork and phloem, as well as stronger xylem vessel staining intensity compared to Daqing germplasm. Regarding various component contents, Yulin germplasm contained significantly higher levels of ethanol-soluble extracts, total saponins, and saikosaponins A, B2, C, D, while Daqing germplasm had significantly higher levels of hemicellulose, starch, and liposoluble extracts. After introduction to Yulin, the Daqing germplasm(YLDQC3) showed increased starch, water-soluble polysaccharides and liposoluble extracts contents, decreased cell wall component content, but no significant difference in other component contents. Metabolomics revealed that saponins and terpenes accumulated significantly in Yulin germplasm, while alcohols and aldehydes accumulated predominantly in Daqing germplasm. Transcriptomics indicated similar gene expression patterns within the same germplasm but specificity between different germplasms. Integrative metabolomic-transcriptomic analysis identified 145 potential key genes associated with the saikosaponin biosynthesis pathway, including one acetyl-coenzyme A(CoA) acetyltransferase gene(ACAT), one 3-hydroxy-3-methylglutaryl-coenzyme A synthase gene(HMGS), two hydroxymethylglutaryl-CoA(HMG-CoA) reductase genes(HMG), one phosphomevalonate kinase gene(PMK), one 1-deoxy-D-xylose-5-phosphate synthase gene(CLA), one hydroxymethylbuten-1-aldol synthase gene(HDR), two farnesyl pyrophosphate synthase genes(FPPS), one squalene synthase gene(SQS), one β-amyrin synthase gene(BAS), 102 cytochrome P450(CYP450) gene family members, and 32 uridine diphosphate-glucuronosyltransferase(UGT) gene family members. ConclusionAmong the three cultivated types, YLC3 most closely resembles YLW in appearance, microscopic features, contents of major bioactive constituents, metabolomic and transcriptomic profiles. Yulin germplasm exhibits superior saponin synthesis capability compared to Daqing germplasm, and Yulin region is more suitable for the growth of B. scorzonerifolium. Based on these findings, it is recommended that artificial cultivation in northern Shaanxi and similar regions utilize the local Yulin germplasm source cultivated for at least three years.
2.Comparison of Wild and Cultivated Polygalae Radix Based on Traditional Quality Evaluation
Yihan WANG ; Yanmeng LIU ; Huaizhu LI ; Yongliang LI ; Shan GUAN ; Zhilai ZHAN
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(5):215-224
ObjectiveBased on the traditional quality evaluation methods summarized in previous dynasties, this paper systematically contrasted the quality differences between wild Polygalae Radix(WPR) and cultivated Polygalae Radix(CPR) from the aspects of character, microscope and chemical composition by modern scientific and technological means, providing a basis for high-quality production and quality control. MethodsCPR and local WPR in Yulin city, Shaanxi province from 1 to 6 years were collected, and a systematic comparative analysis was conducted using traditional pharmacognosy research methods combined with modern multi-omics analysis techniques, including character traits(length, weight, diameter), cross-sectional microscopic features(proportions of cork, phloem, xylem, etc), cell wall component content(hemicellulose, cellulose, lignin), extracts content(water-soluble extract and alcohol-soluble extract), carbohydrate content(starch, water-soluble polysaccharides), contents of total flavonoids, total saponins and specific marker compounds(3,6′-disinapoyl sucrose, polygalaxanthone Ⅲ, tenuifoliside A, tenuifoliside C, sibiricose A5 and A6) and other indexes. Ultra-high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS) was employed to conduct comparative analysis of secondary metabolites in WPR and CPR, and multivariate statistical analysis such as principal component analysis(PCA) and orthogonal partial least squares-discriminant analysis(OPLS-DA) were combined to screen the key differential components of them. ResultsIn terms of appearance, there were significant differences between WPR and CPR. The characteristics of WPR conformed to the "thick wrinkles on the epidermis" recorded in ancient books, featuring a wrinkled surface and grayish-brown appearance. However, CPR had a finer texture and a yellowish white appearance, with weight, length, and diameter increasing with longer cultivation periods. In terms of microscopy, WPR exhibited a thick cork layer with fissures in the phloem, whereas CPR had a thinner cork layer with uniformly arranged cork cells. Younger PR specimens showed numerous phloem fissures in cross-sections, while older specimens display progressively denser arrangements of phloem parenchyma cells. In terms of the contents of various major components, the contents of water-soluble extract, starch and total saponins in WPR were inversely proportional to the root diameter, while the contents of water-soluble extract, water-soluble polysaccharides and total saponins in CPR decreased with the increase of planting years. The content of xanthones in WPR was significantly higher than that of CPR, while the contents of other major components showed no significant change pattern. Among the six indicator components, the average content of sibiricose A5 in WPR was significantly higher than that of CPR, followed by slightly higher content of tenuifoliside A. In CPR, the relative content of 3,6′-disinapoyl sucrose and tenuifoliside A was the highest. The former showed an increase in volatility with increasing cultivation years, while the latter showed a decrease in volatility. The results of differential compound analysis based on UPLC-Q-TOF-MS showed that there were significant differences in metabolites between WPR and CPR samples. Among them, the seven compounds with the largest differences among WPR samples of different thicknesses were polygalasaponins, and for CPR with different planting years, the main differential compounds were oligosaccharide esters. ConclusionThere are differences between WPR and CPR in character, microscopic structure and chemical composition, and some components are inversely proportional with the increase of diameter and cultivation duration due to the distribution characteristics. However, the longer the cultivation years of PR, the closer it is to the "thick wrinkles on the epidermis" of WPR, which has been respected by generations. It is suggested that this traditional character combined with modern component contents should be used as the index of artificial cultivation and quality control of PR.
3.Animal Experiment of 5-FU Slowing-Release Microballoons Given Intraperitoneal
Huaizhu LIANG ; Ye LI ; Shibin ZHU
Chinese Journal of Bases and Clinics in General Surgery 2008;0(10):-
Objective To study the medicine dynamics, distribution in tissue and abdominal cavity fluid concentration of 5-FU after giving intraperitoneal by using a gelatin carrier to be made 5-FU slowing-release microballoons. Methods 5-FU slowing-release microballoons medicine release speed, tissue distributing and the concentration in abdominal cavity fluid were measured by high performance liquid chromatography. Results 5-FU wrapped by gelatin were slowly released. The concentration in abdominal cavity fluid was obviously higher than that in tissue or in blood. Using established standard curve line, it was proved that in body area under curve (AUC) of 5-FU slowing-release microballoons group was obviously higher than that of simple 5-FU injection group, analyzed by 3p97 pharmacokinetic software management. Conclusion 5-FU enwrapped by gelatin can retain an effective anticancer activity concentration in abdominal cavity 7 days after giving intraperitoneal and it is distributed mostly in abdominal cavity.
4.Anti-human platelet tetraspanin (CD9) monoclonal antibodies induce platelet integrin αⅡbβ3 activation in a Fc receptor-independent fashion
Huaizhu WU ; Jiazeng LI ; Lin PENG ; Hanzhi LIU ; Wenjie WU ; Yuling ZHOU ; Qingming HOU ; Dehong KONG
Chinese Medical Journal 2001;114(1):14-18
Objectives To characterize the activation of platelet integrin αⅡbβ3 induced by two anti-human platelet tetraspanin monoclonal antibodies (mAbs), HI117 and SJ9A4, and investigate their potential mechanism of action. Methods Using 125 I-labeled human fibrinogen (Fg), specific Fg binding to human platelets induced by HI117 and SJ9A4 was measured. Results HI117 and SJ9A4 (10?μg/ml and 20?μg/ml) induced specific Fg binding to human platelets, suggesting that the two mAbs evoked activation of platelet integrin αⅡbβ3. Further study indicated that HI117 and SJ9A4 induced integrin αⅡbβ3 activation independent of platelet Fc-receptors, and that HI117 and SJ9A4-induced integrin αⅡbβ3 activation was inhibited by pretreatment of platelets with sphingosine, aspirin, apyrase, and/or PGI2. Conclusions Anti-platelet tetraspanin (CD9) mAbs, HI117 and SJ9A4, can induce platelet integrin αⅡbβ3 activation independent of Fc-receptors. Three signaling pathways, namely thromboxane, secreted ADP, and cAMP pathways, may be involved in the process, with protein kinase C activation presumably being the common step of the three pathways.
5.ANTI-HUMAN PLATELET TETRASPANIN (CD9) MONOCLONAL ANTIBODIES INDUCE PLATELET INTEGRIN αlbβ3 ACTIVATION IN AFc RECEPTOR-INDEPENDENT FASHION
Huaizhu WU ; Jiazeng LI ; Lin PENG ; Hanzhi LIU ; Wenjie WU ; Yuling ZHOU ; Qingming HOU ; Dehong KONG
Chinese Medical Sciences Journal 2000;15(3):145-149
Objective. This study characterized the activation of platelet integrin aⅡbβ3 induced by two anti-human platelet te-traspanin monoclonal antibodies(mAbs),HI117 and SJ9A4. Methods. Using 125I-labeled human fibdnogen(Fg), specific Fg binding to human platelets induced by HI117 and SJ9A4 was measured as indication of activation of platelet integrin αⅡbβ3 by the two mAbs. Results. H1117 and SJ9A4( 10μg/ml and 20μg/ml) induced evident specific Fg binding to human platelets, sug-gesting that the two mAbs evoked activation of platelet integrin αⅡbβ3. Further study indicated that HI117 and SJ9A4 induced integrin t Ⅱ 1β3 activation independent of platelet Fc-receptors, and that HI117 and SJ9A4-induced integrin αⅡbβ3 activation was inhibited by sphingosing, aspirin, apyrase, and/or PGI2. Conclusion. The anti-platelet tetraspanin(CD9)mAbe,HI117 and SJ9A4, can induce platelet integrin αⅡbβ3 act-vation independent of Fc-receptors. Three signaling pathways,i.e.thromboxane,secreted ADP, and cAMP pathways may be involved in the process, with protein kimse C activation presumably being the comtmon step of the three pathways.

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