Study on the effect of natural compound XCQ-9 of Cynanchum paniculatum on the proliferation and apoptosis of Jurkat cell and its mechanism / 中国药房

OBJECTIVE To investigate the inhibitory effect of natural compound XCQ-9 of Cynanchum paniculatum on the proliferation and apoptosis of Jurkat cell line of human T-cell acute lymphoblastic leukemia and its possible mechanism. METHODS Jurkat cell was used as the leukemia cell model， and MTT assay was adopted to detect the inhibitory effects of 0 （blank control）， 2.5， 5， 10， 20 and 40 μmol/L XCQ-9 on the proliferation of Jurkat cell after treated for 24， 48， 72 h. After treated with 0 （blank control）， 2.5， 5， 10 μmol/L XCQ-9 for 24 h and 48 h， the cell cycle and apoptosis were analyzed by flow cytometry. The expressions of Caspase-9， Cleaved Caspase-9， Caspase-3， Cleaved Caspase-3， poly ADP-ribose poly-merase （PARP）， Cleaved-PARP， cyclin-dependent kinase 1 （CDK1） and Cyclin B1 were detected by Western blot after treated for 24 h. RESULTS Compared with blank control group， XCQ-9 at different concentrations could significantly decrease the survival rate of Jurkat cells （P＜0.01）， and showed a dose and time-dependent manner. After 48 h treatment of 5， 10 μmol/L XCQ-9， Jurkat cell apoptosis was induced significantly （P＜0.05 or P＜0.01）， and the cell was arrested in G2 phase （P＜0.01）. After 24 h treatment of 10 μmol/L XCQ-9， the protein expressions of CDK1 and Caspase-9 were remarkably down-regulated （P＜0.01）， while the protein expressions of Cyclin B1， Cleaved Caspase-9， Cleaved Caspase-3 and Cleaved PARP were significantly up-regulated （P＜0.05 or P＜0.01）. CONCLUSIONS XCQ-9 plays anti-tumor effect through inducing G2 phase arrest to inhibit proliferation and 5008） activating Caspase pathway to increase apoptosis.

Effects of Achyranthis bidentatae -Cynanchum otophyllum on oxidative stress of cerebral tissue in Parkinson ’s disease mice with syndrome of hyperactivity of liver -Yang / 中国药房

OBJECTIVE To explore the effects of Achyranthis b identatae-Cynanchum otophyllum as core couplet medicinals of Zhengan xifeng decoction on oxidative stress of cerebral tissue in Parkinson ’s disease （PD）mice with syndrome of hyperactivity of liver-Yang. METHODS C57BL/6 mice were randomly divided into normal control group ，model group ，Zhengan xifeng decoction group，A. bidntatae group，C. otophyllum group and couplet medicinals of A. bidentatae-C. otophyllum group，with 10 mice in each group. PD model of hyperactivity of liver -Yang was established by intragastric administration of Aconitum carmichaelii decoction（4 g/kg） and intraperitoneal injection of 1-methyl-4-phenyl-1，2，3，6-tetrahydropyridine （25 mg/kg）. The behavioral changes and ultrastructure of substantia nigra neurons in mice were observed . The expressions of tyrosine hydroxylase （TH）positive neurons in substantia nigra were detected . The expressions of total antioxidant capacity （T-AOC），superoxide dismutase （SOD） and malondialdehyde（MDA）in substantia nigra as well as mRNA and protein expressions of nuclear factor -erythroid 2-related factor 2 （Nrf2），thioredoxin reductase 1（Trxr1），thioredoxin interacting protein （Txnip）were also determined . RESULTS Compared with model group ，PD behavior and ultrastructure of substantia nigra neurons were all improved in administration groups . The expressions of TH positive neurons ，T-AOC（except for C.otophyllum group）and SOD （except for C.otophyllum group），mRNA relative expression and protein expression levels of Nrf 2 and Trxr 1 were all increased significantly ；the expression of MDA （except for C.otophyllum group）and mRNA relative expression and protein expression levels of Txnip were all decreased significantly （P＜0.05）. Intervention effect of couplet medicinals of A. bidentatae-C. otophyllum group was better than that of A.bidntatae group and C. otophyllum group（P＜ 0.05），and the effect was similar to that of decoction group （P＞0.05）. CONCLUSIONS The couplet medicinals of A. bidentatae-C. otophyllum can inhibit the level of oxidative stress in the cerebral tissue of PD mice with hyperactivity of liver -Yang by targeting Nrf 2，and play a protective role on the brain neurons . Its effect is better than that of A.bidntatae and C.otophyllum，and it plays the same role as that of the formula in tonifying the kidney ，softening the liver and suppressing the Yang .

Establishment of fingerprint ，analysis of chemical pattern recognition and multi-index content determination of Cynanchum auriculatum / 中国药房

OBJECTIVE To establis h the fingerprint of Cynanchum auriculatum，to conduct its chemical pattern recognition analysis，and to determine the contents of four components at the same time. METHODS High performance liquid chromatography （HPLC）method was adopted. The determination was performed on ACE UExcel C 18 column with mobile phase consisted of acetonitrile-0.1％ phosphoric acid solution （gradient elution ）at the flow rate of 1.2 mL/min. The determination wavelength was set at 210 nm，and the column temperature was 40 ℃. The sample size wa s 10 μL. Taking qingyang shengenin as the reference ，HPLC fingerprints of 16 batches of C. auriculatum medicinal materials were drawn and similarity was evaluated by using the Similarity Evaluation of Chromatographic Fingerprints of Traditional Chinese Medicine （2012 edition）， and the common peaks were determined. SPSS 26.0 software andSIMCA 14.0 software were used for cluster analysis ，principal component analysis and orthogonal partial least squares- discriminant analysis. The differential components affecting the quality of C. auriculatum were screened by taking the value of variable importance in projection （VIP）greater than 1 as the standard ；same HPLC method was used to determine the contents of syringic acid ，acyl asclepiadelenin ，baishouwubenzophenone and qingyang shengenin. RESULTS There were 29 common peaks in 16 batches of C. auriculatum ，with a similarity of 0.723-0.998. Four common peaks were identified ，namely syringic acid （peak 7），acyl asclepiadoidin （peak 9），baishouwubenzophenone（peak 13）and qingyang shengenin（peak 15）. The results of cluster analysis showed that 16 batches of C. auriculatum could be clustered into three categories ，among which S 1 were grouped into one category，S3 were grouped into one c ategory，S2，and S 4-S16 were grouped into one category. The results of principal component analysis showed that the cumulative variance contribution rate of the five principal components was 88.706％，and the classification results were consistent with the results of cluster analysis. The results of orthogonal partial least squares-discriminant analysis showed that the common peaks （from large to small ）with VIP value greater than 1 were peak 20，peak 10，peak 25，peak 12， peak 15（qingyangshengenin），peak 21，peak 14，peak 16，peak 26，peak 22 and peak 17. The linear ranges of syringic acid，acyl asclepterin，baishouwubenzophenone and qingyangshengenin were 0.715 3-45.778 0，2.379 4-152.281 0，0.642 0- 41.085 0，14.541 6- 930.662 0 µg/mL respectively （all R 2＞0.999）. The quantitative limits were 0.357 7，0.475 9，0.642 0 and 2.423 6 μg/mL；the detective limits were 0.146 0，0.164 1，0.248 8 and 0.833 3 μg/mL，respectively. RSDs of precision ，repeatability and stability （24 h）tests were less than 3％；the average recoveries were 99.11％（RSD＝2.00％，n＝9），98.54％（RSD＝2.21％，n＝9）， 96.33％（RSD＝2.54％，n＝9）and 95.96％（RSD＝2.93％，n＝9）；the contents were 17.12-147.80，95.23-583.10（S8 below the quantitative limit ），16.91-210.88 and 211.68-3 587.15（S1 below the quantitative limit ）μg/g，respectively. CONCLUSIONS Established HPLC fingerprint and the method of content determination are stable ，reliable，accurate and reproducible. Combined with analysis of chemical pattern recognition ，it can be used for the quality control of C. auriculatum .

Identification,biological characteristics and fungicide screening of pathogen of southern blight in Cynanchum stauntonii / 中国中药杂志

During the high-temperature and rainy season from June to October in 2017-2019,serious southern blight broke out in the Cynanchum stauntonii planting area in Tuanfeng county,Hubei province,which had a great impact on the yield and quality of medicinal materials. In this study,the pathogen of C. stauntonii was isolated,purified,and identified,and the pathogenicity was tested according to Koch's postulates. Meanwhile,the biological characteristics of the pathogen were analyzed. On this basis,the effective fungicides were screened in laboratory. Finally,the pathogen( BQ-1) was identified as Athelia rolfsii( Deuteromycotina,Basidiomycota,anamorph: Sclerotium rolfsii). The optimum growth conditions for BQ-1 were 25-30 ℃,p H 5-8,and alternating light and dark.The effective chemical fungicides were lime-sulphur-synthelic-solution( LSSS) and flusilazole,and the effective botanical fungicide was osthole. BQ-1 was highly homologous to the pathogen HS-1 of peanut southern blight,with the similarity of 18 S r DNA and TEF sequences at 99. 09%. The southern blight in C. stauntonii might be resulted from that in peanut. In the production of C. stauntonii,the following measures should be taken: avoiding rotation or neighboring with peanut,draining water from June to October to reduce humidity,and reasonably applying fungicides.

Effect of C21 steroidal glycoside of Cynanchum auriculatum on liver and kidney fibrosis through TLR4 pathway / 中国中药杂志

The liver and kidney fibrosis model was established by thioacetamide(TAA) and unilateral ureteral obstruction(UUO) in SD rats. The rats were randomly divided into three groups: model group, low and high-dose groups of C21 steroidal glycosides of Cynanchum auriculatum. Another blank control group was set. Four weeks later, serum was taken to detect the biochemical indexes of liver and kidney function. Urine protein and urine creatinine were detected by kits. Liver and kidney tissue samples were stained with HE and Masson staining, and hydroxyproline content was detected. Western blot was used to detect expressions of fibrotic proteins, inflammatory factors and TLR4 signaling pathways, so as to observe the preventive and therapeutic effects of C21 steroidal glycosides from C. auriculatum on hepatic and renal fibrosis and explore its molecular mechanism. Four weeks later, serum biochemical results showed that liver and kidney functions were seriously damaged, and pathological sections showed that inflammatory cell infiltration, decrease of parenchymal cells, and increase of interstitial fibrosis in liver and kidney tissues. The results showed that low and high doses(150, 300 mg·kg~(-1)) of C21 steroidal glycosides could significantly reduce the collagen deposition and the pathological changes of liver and kidney fibrosis compared with the model group. At the same time, we found that the expression levels of TLR4 and MyD88 signaling pathway proteins were significantly increased in the liver and kidney tissues of the model group, and a large number of NF-κB signaling pathway proteins migrated into the nucleus. On the contrary, the expression levels of TLR4, MyD88 signaling pathway proteins and the nuclear migration of NF-κB were significantly inhibited in the low and high dose groups of C21 steroidal glycosides from C. auriculatum. Therefore, it was speculated that the mechanism of C21 steroidal glycoside for preventive and therapeutic effect on hepatic and renal fibrosis was related to inhibit TLR4/MYD88/NF-κB inflammatory pathway, thus preventing hepatic and renal fibrosis.

Immunological Activity and Chemical Constituents of Fermentation Product in Cynanchum auriculatum Royle ex Wight / 中国药学杂志

OBJECTIVE: To study immunological activity and chemical constituents of fermentation product of the roots of Cynanchum auriculatum Royle ex Wight. METHODS: The effect of fermentation product of the roots of C. auriculatum on the spleen index and thymus index of the immunocompromised rat was measured by weighing and phagocytosis of chicken red blood cells, silicagel, Sephadex LH-20 and recrystallization were used for isolation and purification.The structures are identified by physicochemical properties and spectral data analysis, and the separated compounds were identified by HPLC. RESULTS: The fermentation product of the roots of Cynanchum auriculatum Royle ex Wight auricular leaf can improve the spleen and thymus index of mice and is superior to the alcohol extract of Cynanchum auriculatum Royle ex Wight. The results of phagocytosis of chicken red blood cells by peritoneal macrophages in mice are positive. Eleven compounds Compounds were isolated from the fermentation product in the roots of Cynanchum auriculatum Royle ex Wight, which were identified as 2,4-dihydroxyacetophenon(1), 2,5-dihydroxyacetophenone(2), baishouwubenxophenone(3), caudatin(4), kidjolanin(5), periplogenin(6),gallic acid(7),oleanolic acid(8), deacylmetaplexigenin(9),sorbitol(10) and linoleic acid(11). CONCLUSION: The fermentation product of the roots of Cynanchum auriculatum Royle ex Wight have the effect of improving immune function.Compounds 7,10,11 are the substances produced during the fermentation process.

Effects of total C-21 steroidal glucosides from Cynanchum auriculatum on renal fibrosis in rats and mechanisms involved / 中草药

Objective: To investigate the effects of C-21 steroidal glucosides from Cynanchum auriculatum on renal fibrosis in rats caused by unilateral ureteral ligation and explore its mechanisms. Methods: A total of 15 rats were randomly chosen as sham operation group (Sham), while the remaining rats underwent unilateral ureteral ligation. The rats after the operation were randomly divided into four groups, namely the Sham operation group, the model group, the positive control group (20 mg/kg), C. auriculatum high group (400 mg/kg), C. auriculatum low group (200 mg/kg), 15 per group, and the rats in each group were administrated with corresponding drugs. After the intervention of 28 d, all the rats were sacrificed and the kidneys were then removed. The content of hydroxyproline was measured. HE and Masson staining were conducted to assess kidney pathological changes and renal fibrosis. The protein expression of TGF-β1, α-SMA, and E-cadherin were tested with immunohistochemistry and Western blotting. The mRNA expression of collagen-I and collagen-III was evaluated using qRT-PCR. Results: Compared with the model group, C-21 steroid glycosides significantly alleviated the kidney pathological injury and renal fibrosis, and reduced fibrous tissue and collagen proliferation. C-21 steroid glucosides from C. auriculatum can significantly reduce the kidney/body weight ratio and the content of hydroxyproline in kidney tissue (P < 0.05, 0.01), and it showed in a dose-dependent manner. Compared with the model group, the expression of α-SMA and TGF-β1 was decreased significantly, and the expression of E-cadherin was increased significantly (P < 0.05, 0.01) in C. auriculatum group. Moreover, compared with the model group, C-21 steroidal glucosides significantly down-regulated the mRNA expression of collagen-I and collagen-III mRNA (P < 0.01). Conclusion: C-21 steroid glycosides from C. auriculatum can effectively attenuate renal fibrosis in rats with unilateral ureteral ligation and its underlying mechanisms may be related to inhibiting the over-expression of collagen-I and collagen-III, down-regulating the expression of α-SMA and TGF-β1, and up-regulating the expression of E-cadherin. By regulating the tubular epithelial-mesenchymal transition, it exerts its anti-fibrotic effect.

A comparative study on antidiarrheal effects of Mongolian medicines Holarrhena antidysenterica, Forsythia suspensa and Cynanchum thesioides / 中草药

Objective: To compare the antidiarrheal effects of Mongolian medicine, Holarrhena antidysenterica, Forsythia suspensa and Cynanchum thesioides on diarrhea model rats and investigate its effects on serum DAO (diamine oxidase), cAMP (cyclic adenosine phosphate), TNF-α (tumor necrosis factor-α), ATPase and calcium ions. Methods: The normal control group, model group, H. antidysenterica low-dose and high-dose groups, F. suspensa low dose and high-dose groups, C. thesioides low dose and high dose groups were set. Except the normal control group, the other groups were ig administrated water decoction of Cassia angustifolia to establish diarrhea model; After the success of the model, the rats in treatment groups were administrated by gastric drug for 7 d, the type mental state, diarrhea and body weight changes were observed. the abdominal aortic blood was obtained at the last day of fasting 12 h after the administration. DAO, cAMP, TNF-α, ATP enzyme, OD value of the calcium ions in serum were determined by using ELISA (enzyme-linked immune detection reagent) kits. Results: General status: except for the normal group, the mental state of the rats in the other groups was depressed after modeling, the fur color of them was significantly decreased and the body weight was decreased. The diarrhea rate was 100% on the 4th day after modeling. Compared with the model group, there were significant differences in the number of loose stools, grade of loose stools and diarrhea index (P < 0.05, 0.01) in each administration group. The serum DAO, TNF-α, ATPase, cAMP and calcium ion OD values were compared: the serum concentrations of DAO in H. antidysenterica low dose group, C. thesioides low dose group and F. suspensa high and low dose groups of rats were significantly lower than model group with significant differences (P < 0.05), and were significantly higher than normal group. The serum concentration of TNF-α in C. thesioides high and low dose groups were lower compared with model group (P < 0.05). The serum ATPase in C. thesioides high and low dose groups had significant difference (P < 0.05) compared with model group. The serum concentration of cAMP in H. antidysenterica high-dose group and F. suspensa low-dose group was significantly lower compared with model group with significant differences (P < 0.05). Serum Ca2+ concentration in the drug administration groups was significantly different from that in the model group (P < 0.05). Conclusion: The antidiarrheal effect of C. thesioides is better than that of H. antidysenterica and F. suspensa.

Study on the Quality Standards of Mongolian Medicine Cynanchum thesioides / 中国药房

OBJECTIVE：To establish the quality standards of Mongolian medicine Cynanchum thesioides. METHODS ：TLC was used for the qualitative identification of C. thesioides . According to 2015 edition of Chinese Pharmacopeia （part Ⅳ），the moisture，total ash and ethanol-soluble extract were determined. HPLC method was used to determine the content of thesioideoside in C. thesioides . RESULTS ：TLC spots were clear ，there were same yellow green fluorescent spots on the corresponding position of the sample （C. thesioides ）and control （thesioideoside）. In 22 batches of samples ，contents of moisture were 6.18％-12.97％，total ash were 4.64％-7.95％，ethanol-soluble extract were 12.46％-32.70％. The linear range of thesioideoside were 0.048-3.050 μg（R2＝ 0.999 9）. RSDs of precision ， stability， repeatability and durability tests were all less than 1％ . The recoveries were 104.03％-106.36％（RSD＝0.96％，n＝6）. The contents of thesioideoside in 22 batches of C. thesioides were 0.006 2％-0.130 5％. CONCLUSIONS：It is suggested that the moisture and total ash should not exceed 11.50％ and 7.50％，respectively；the contents of ethanol-soluble extract and the sioideoside are no less than 17.00％ and 0.05％，respectively. The established quality standards can be used for quality control of Mongolian medicine C. thesioides .

Research progress on Cynanchi Bungei Radix / 中草药

Cynanchi Bungei Radix is a traditional Chinese herbal medicine which has been used as a tonic agent owing to its primary abilities of tonifying liver and kidney, nourishing blood and replenishing semen, strengthening tendons and bones, promoting hair growth, and prolonging life. Pharmacological effects and material basis of Cynanchi Bungei Radix have been extensively investigated. This paper reviews the recent research progress of Cynanchi Bungei Radix based on the difference in the pharmacological effects and content of different chemical constituents in different species of Cynanchi Bungei Radix. It provides a scientific basis for further rational application of the medicinal value of different varieties of Cynanchi Bungei Radix.

Effects of total C-21 steroidal glucosides from Cynanchum auriculatum on oxidative stress pathway in mice with liver injury / 中国中药杂志

The study aimed to investigate the mechanism of hepatoprotective effect of C-21 steroidal glucosides from Cynanchum auriculatum( Baishouwu) on oxidative stress in mice with liver injury. Mice were randomly divided into normal group,model group,positive control group,Baishouwu high group and Baishouwu low group. The liver injury model was induced by intraperitoneal injection of CCl4 peanut oil solution. All mice were sacrificed to collect blood and liver specimens. The activities of serum levels of ALT and AST were detected. The content of MDA and the activity of SOD in liver homogenate were examined by colorimetry method. Tissues were stained with hematoxylin-eosin for histological examination. The hepatic protein expressions of NF-κB p65,p-IκBα,i NOS and COX-2 were detected by Western blot. The mRNA expressions of TNF-α and IL-6 were determined by RT-PCR. It was found that treatment with C-21 steroidal glucosides from Baishouwu successfully attenuated liver injury induced by CCl4,as shown by decreased levels of serum biochemical indicators( AST,ALT)( P<0. 01). Administration of total C-21 steroidal glucosides enhanced the activity of SOD( P<0. 01) and decreased the content of MDA( P<0. 01) in liver homogenate. Microscopic features suggested that treatment with C-21 steroidal glucosides from Baishouwu was effective in inhibiting CCl4-induced hepatocyte edema and degeneration. Further studies showed that NF-κB p65 overexpression induced by CCl4 was decreased by C-21 steroidal glucosides,leading to the markedly down-regulated protein expression levels of p-IκBα,i NOS and COX-2,as well as the depression of TNF-α and IL-6 mRNA expressions. In conclusion,total C-21 steroidal glucosides from Baishouwu exhibited potent effect on oxidative stress pathway in mice with liver injury induced by CCl4,with enhanced activity of SOD,decreased content of MDA,and down-regulated levels of NF-κB p65,p-IκBα,i NOS and COX-2.

Chemical constituents in water fraction of Cynanchum auriculatum / 中草药

Objective To investigate the chemical constituents of Cynanchum auriculatum. Methods Compounds were isolated and purified by using a combination of various chromatographic techniques including D-101 macroporous adsorptive resins, silica gel, Sephadex LH-20, and other methods. Their structures were elucidated by MS, NMR, and other modern spectroscopies. Results Fifteen compounds were isolated and identified as isocopoletin (1), isofraxidin (2), deacetylmetaplexigenin (3), vomifoliol (4), 4,4-dimethyl heptanedioic acid (5), (+)-isolariciresinol (6), 3-hydroxypyridine (7), 3-hydroxy-2-methylpyridine (8), 5-hydroxyl-2-hydroxymethylpyridine (9), 2-methyl-6-(2’,3’,4’-trihydroxybutyl)-pyrazine (10), kiwiionol (11), picein (12), adenosine (13), cynanoneside B (14), and cynanoneside A (15). Conclusion Compounds 1-2, 4-13 are isolated from the plants of genus Cynanchum Linn for the first time, while compound 15 is firstly obtained from C. auriculatum.

Molecular identification of Cynanchum auriculatum and its closely related species based on ITS2 sequence / 中草药

Objective To identify Cynanchum auriculatum and its closely related species using the ITS2 barcode. Methods A total of 54 samples of C. auriculatum and its related species were collected. Genomic DNAs were extracted from these samples. The ITS2 sequences of these samples were amplified and bidirectional sequenced by PCR. The obtained sequences were submitted to the Gen Bank and the ITS2 sequences of 47 samples belonging to 15 species were downloaded from the GenBank, and ITS2 sequences were annotated by ITS2 database. A total of 101 ITS2 sequences were aligned and the intraspecific and interspecific distances were calculated using the MEGA 5.0. Identification analyses were performed using the similarity search method and nearest distance method, and were presented intuitively by constructing neighbor-joining (NJ) tree. Results The length of all ITS2 sequences of C. auriculatum was 249 bp, which was a haplotype and was close to Cynanchum. There was a significant difference between the interspecific and intraspecific genetic distances of the ITS2 sequences. The NJ tree showed that C. auriculatum obviously differed from its closely related species, which showed high monophyly. According to the secondary structure of ITS2, it was also possible to distinguish between C. auriculatumi and Asclepiadaceae related species. Conclusion As a DNA barcode, ITS2 sequences can stably and accurately distinguish C. auriculatum from its closely related species and also provide a new technique to ensure the clinical safety in utilization of Chinese materia medica.

Two new C steroidal glycosides isolated from Cynanchum komarovii / 中国天然药物

The present study was designed to further investigate the C steroidal glycosides in Cynanchum plants. Two new steroidal glycosides based on a 13, 14:14, 15-disecopregnane-type aglycone, komaroside P (1) and komaroside Q (2), together with three known compounds (3-5) were isolated from the whole herbs of Cynanchum komarovii. The aglycones of compounds 1 and 2 were two new disecopregnane. Their structures were elucidated on the basis of 1D, 2D NMR spectroscopic data and acid hydrolysis. All the compounds (1-5) showed potent inhibitory activities against human leukemia cell lines (HL-60) with IC values ranging from 16.6 to 26.3 μmol·L, compared to the positive control 5-fluorouracil (6.4 μmol·L).

Two new C steroidal glycosides isolated from Cynanchum komarovii / 中国天然药物

The present study was designed to further investigate the C steroidal glycosides in Cynanchum plants. Two new steroidal glycosides based on a 13, 14:14, 15-disecopregnane-type aglycone, komaroside P (1) and komaroside Q (2), together with three known compounds (3-5) were isolated from the whole herbs of Cynanchum komarovii. The aglycones of compounds 1 and 2 were two new disecopregnane. Their structures were elucidated on the basis of 1D, 2D NMR spectroscopic data and acid hydrolysis. All the compounds (1-5) showed potent inhibitory activities against human leukemia cell lines (HL-60) with IC values ranging from 16.6 to 26.3 μmol·L, compared to the positive control 5-fluorouracil (6.4 μmol·L).

Chemical constituents from endophytic fungus Plectosphaerella cucumerina YCTA2Z1 of Cynanchum auriculatum / 中草药·英文版

Objective: To study the chemical constituents from the EtOAc extract of endophytic fungal Plectosphaerella cucumerina YCTA2Z1. Methods: The chemical constituents were isolated and purified by silica gel column, Sephadex LH-20, and reverse-phase C-18 column chromatography as well as crystallization. Results: Thirteen compounds were isolated from the EtOAc extract of the fungal strain YCTA2Z1. Their chemical structures were elucidated according to the spectral evidence. They were identified as caudatin (1), baishouwubenzophenone (2), cynandione B (3), asterbatanoside A (4), p-hydroxyphenethyl-O-β-D-glycoside (5), caffeic acid (6), ferulic acid (7), 2,5-dihydroxyacetophenone (8), protocatechuic acid (9), vanillic acid (10), stearic acid (11), azelaic acid (12), and succinic acid (13). Conclusions: It is the first chemical study on endophytic fungi from Cynanchum auriculatum and all the compounds are obtained from the species, the genus, as well as the family Plectosphaerellaceae for the first time.

Pharmacognostical study of Cynanchum stauntonii and Cynanchum glaucescens, botanical sources of TCM Baiqian / 中草药·英文版

Objective: Cynanchum stauntonii and Cynanchum glaucescens are botanical species of Baiqian (Cynanchi Stauntonii Rhizoma et Radix) in Chinese Pharmacopoeia, in which, however, there are no microscopic identification. Therefore, we provided the morphological and microscopic identification of the crude drug for updating Chinese Pharmacopoeia. Methods: Twelve batches of C. stauntonii and three batches of C. glaucescens and their crude drugs were taxonomically, morphologically, and microscopically examined. Results: Taxonomically, C. stauntonii had narrowly lanceolate leaves with acuminate apex and 5mm long petiole; Whereas C. glaucescens was oblong-lanceolate or oblong with rounded or acute apex in leaves, and had very short or no petiole. Morphologically, rhizomes of C. stauntonii and C. glaucescens both had hollow pith, but the hollow pith occupied about a half of the rhizome's diameter in C. stauntonii, whereas only a very small proportion of the overall diameter in C. glaucescens. Moreover, microscopic observation showed the difference in the proportion of xylem and in rhizome transverse-sections of the two species along with the difference in the size of the pith. Finally, laticifers and rhizome epidermal secretory cells were present in the powders of C. stauntonii, but absent from C. glaucescens. Conclusion: Based on observation of morphological and microscopic characteristics, the two species can be distinguished by the size of the pith, proportion of xylem of rhizomes, and crude drug powder characters such as laticifers and secretory cells.

Paniculatumoside G, a new C21 steroidal glycoside from Cynanchum paniculatum

ABSTRACT A new C21 steroidal glycoside, paniculatumoside G, together with neocynapanogenin C isolated for the first time from the natural source and two known compounds were isolated and characterized from the roots and rhizomes of Cynanchum paniculatum (Bunge) Kitag. ex H.Hara, Apocynaceae, a commonly used Traditional Chinese Medicine. On the basis of spectroscopic analysis, including HR-ESI-MS, 1D and 2D NMR spectral data, the structure of the new C21 steroidal glycoside was elucidated as neocynapanogenin H 3-O-β-D-oleandropyranoside.

Content Determination of Paeonol in Cynanchum paniculatum by Three-dimensional Fluorescence Coupled with ATLD Algorithm / 中国药房

OBJECTIVE:To determine the content of paeonol in Cynanchum paniculatum by three-dimensional fluorescence coupled with ATLD algorithm. METHODS:The trilinear model was established. The principle of least square was used for ATLD. The component value was fitted by core consistency diagnosis,and inner filter effect was corrected by mathematical correction method. Fluorescence scanning condition included excitation wavelength of 250-400 nm,emission wavelength of 410-600 nm, wavelength interval of 5 nm,slit width of 5.0/5.0 nm,scanning speed of 1200 nm/min. Determination and absorption spectrum condition included scanning wavelength of 250-600 nm,scanning speed of 600 nm/min;Al(Ⅲ)was used as the sensitizer to in-crease the fluorescence intensity of paeonol. The content of paeonol was determined by HPLC. RESULTS:The linear range of pae-onol was 0.132-1.188 μg/mL(r=0.9999). RSDs of precision,stability and reproducibility tests were less than 3.0%. The recover-ies were 100.1%-104.7%(RSD=2.39%,n=6). The analysis spectrum of paeonol was almost completely overlapped with the actu-al spectrum. The results of HPLC method are very similar to those of ATLD algorithm. CONCLUSIONS:The three-dimensional fluorescence coupled with ATLD algorithm is simple,rapid,efficient and accurate,and can be used for the qualitative and quantita-tive analysis of complex system.

Recent advances in phytochemistry and pharmacology of C21 steroid constituents from Cynanchum plants / 中国天然药物

Cynanchum is one of the most important genera in Asclepiadaceae family, which has long been known for its therapeutic effects. In this genus, 16 species are of high medicinal value. The extracts of the root and/or rhizome parts have been applied in traditional Chinese medicines (TCM) for the prevention and treatment of various illnesses for centuries. C21 steroids, as the typical constituents of Cynanchum species, possess a variety of structures and pharmacological activities. This review summarizes the comprehensive information on phytochemistry and pharmacology of C21 steroid constituents from Cynanchum plants, based on reports published between 2007 and 2015. Our aim is to provide a rationale for their therapeutic application, and to discuss the future trends in research and development of these compounds. A total of 172 newly identified compounds are reviewed according to their structural classifications. Their in vitro and in vivo pharmacological studies are also reviewed and discussed, focusing on antitumor, antidepressant, antifungal, antitaging, Na(+)/K(+)-ATPase inhibitory, appetite suppressing and antiviral activities. Future research efforts should concentrate on in vitro and in vivo biological studies and structure activity relationship of various C21 steroid constituents.