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.
Basidiomycota ; Cynanchum ; Fungicides, Industrial/pharmacology* ; Humidity

OBJECTIVETo study the genetic diversity of rDNA ITS sequences in different species of Bai Shouwu, utilize the molecular diversity of ITS sequences to authenticate the different species of Bai Shouwu.

METHODFirstly, total DNA was extracted from the different species of Bai Shouwu. Secondly, the ITS sequence was amplified by PCR with universal primer of ITS and sequenced after cloning and purification.

RESULTFrom four species the complete sequence of ITS and 5.8 S rDNA, the partial sequences of 18S rDNA and 26S rDNA were obtained. The rDNA ITS sequences of Cynanchum bungei (sign in No. GU198970 and No. GU479037) were obtained. Ten variable sites among the sequences were found.

CONCLUSIONITS sequence could be used to authenticate the species. The method could be used to identify germplasm resources and authenticate.

Cynanchum ; classification ; genetics ; DNA, Plant ; genetics ; DNA, Ribosomal Spacer ; genetics ; Molecular Sequence Data

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.
Animals ; Cynanchum ; chemistry ; Humans ; Molecular Structure ; Plant Extracts ; chemistry ; pharmacology ; Steroids ; chemistry ; pharmacology

Eleven C21 steroids were isolated from chloroform extract of roots of Cynanchum otophyllumby silica gel, MCI, ODS columns, and semi-preparative HPLC. Their structures were determined by spectroscopic data analysis as otophylloside B(1), caudatin-3-O-beta-D-cymaropyranosyl-(1-->4)-beta-D-oleandropyranosyl-(1-->4)-beta-D-cymaropyranosyl-(1-->4)-beta-D-cymaropyranoside (2), caudatin-3-O-beta-D-oleandropyranosyl-(1-->4)-beta-D-oleandropyranosyl-(1-->4)-beta-D-cymaropyranosyl-(1-->4)-beta-D-cymaropyranoside (3), caudatin-3-O-beta-D-oleandropyranosyl-(1-->4)-beta-D-digitoxopyranosyl-(1-->4)-beta-D-cymaropyranoside (4), otophylloside O (5), gagamine-3-O-beta-D-oleandropyranosyl-(1-->4)-beta-D-cymaropyranosyl-(1-->4)-beta-D-cymaropyranoside (6), sinomarinoside B (7), mucronatosides C (8), wallicoside J (9), stephanoside H (10), and qinyangshengenin-3-O-beta-D-oleandropyranosyl-(1-->4)-beta-D-cymaropyranosyl-(1-->4)-beta-D-digitoxopyranoside (11). Among them, compounds 2-3, and 6-11 were separated from the roots of this plant for the first time.
Cynanchum ; chemistry ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Magnetic Resonance Spectroscopy ; Molecular Structure ; Plant Roots ; chemistry ; Steroids ; chemistry ; isolation & purification

OBJECTIVETo evaluate the inhibitive effect of C-21 steroidal glycosides from the root of Cynanchum auriculatum (CGB) on rat glioma C6 cells.

METHODSC6 cells were treated with CGB for 24, 48,72 h at concentration of 30, 60, 120 mg/L, respectively. MTT assay was used for evaluating cell viability; fluorescence-activated cell sorting analysis after Annexin V/propidium iodide staining or single propidium iodide staining was used to test cell apoptosis and cell cycle.

RESULTSCGB at 30, 60, 120 mg/L concentration-dependently decreased C6 cell viability (P<0.001). CGB at 60 and 120 mg/L induced C6 cell apoptosis and cell cycle arrest. The fraction of G0/G1 cells was increased (P<0.05) and that of S phase cells was decreased (P<0.01).

CONCLUSIONCGB can inhibit the growth of rat glioma C6 cells, and induce apoptosis and G0/G1 cell cycle arrest.

Animals ; Apoptosis ; drug effects ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cynanchum ; chemistry ; Glioma ; pathology ; Monosaccharides ; pharmacology ; Rats ; Steroids ; pharmacology

OBJECTIVETo study the optimizal hydrolysis process for C21 steroidal glycoside of Bai Shou Wu by acetic acid.

METHODThe effects of acetic acid concentration, reaction temperature and reaction time had been investigated using orthogonal design and the contents of kidjoranin 3-O-beta-digitoxopyranoside, caudatin, kidjoranin 3-O-alpha-L-diginopyranosyl-(1 --> 4)-beta-cymaropyranoside and caudatin 3-O-beta-cymaropyranoside as response indexs were determined by the high performance liquid chromatography.

RESULTThe factors influencing acetic extraction efficiency were as follows: A > B > C (A. reaction temperature; B. reaction time; C. acetic acid concentration). The optimal hydrolysis condition obtained was: refluxing for 6 hours with 5.0% dilute CH3COOH solution at 100 degrees C.

CONCLUSIONThe content of antitumor active ingredients under the optimum hydrolysis condition is raised obviously and has a great part in studying this antitumor drug.

Acetates ; pharmacology ; Cynanchum ; chemistry ; metabolism ; Drugs, Chinese Herbal ; chemistry ; metabolism ; Glycosides ; analysis ; chemistry ; metabolism ; Hydrolysis ; drug effects ; Temperature ; Time Factors

High performance liquid chromatography coupled with on-line electrospray tandem mass spectrometry (HPLC/ESI-MS/MS) was used to identify C21 steroidal glycosides in the roots of Cynanchum atratum. The structures of C21 steroidal glycosides were deduced from mass fragments features in positive and negative mode. The constituents of C. atratum were separated and detected. 7 compounds were identified by comparing their ESI-MS/MS data with the reference compounds and 2 compounds were inferred solely by the ESI-MS/MS data. The method is sensitive, and provides good separation and rapid qualitative characterization of C21 steroidal glycosides in the roots of C. atratum.
Chromatography, High Pressure Liquid ; Cynanchum ; chemistry ; Glycosides ; analysis ; chemistry ; Spectrometry, Mass, Electrospray Ionization ; Steroids ; chemistry ; Tandem Mass Spectrometry

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.
Animals ; Cynanchum ; Fibrosis ; Glycosides ; Kidney/pathology* ; Liver ; NF-kappa B/genetics* ; Rats ; Rats, Sprague-Dawley ; Toll-Like Receptor 4/genetics*

OBJECTIVETo develop an approach to the determination of saponins in Radix Cynanchi Atrati, and to optimize the parameters for purified the preparation of total saponins by macroporous resin column chromatography.

METHODUsing cynanversicoside A as a reference, the determination of saponins was performed; according to the elution rate and the purity of the products, the preparation performance of total saponins by macroporous resin was investigated, and its parameters were optimized.

RESULTThe saponins in Radix Cynanchi Atrati were successfully determined at 518 nm by vanillin-perchloric acid as spray reagent. The macroporous resin HP-20 showed static absorption ratio of 59. 3 mg x g(-1); the 70% ethanol extraction of Radix Cynanchi Atrati was eluted from column of macroporous resin HP-20 by water and 30% ethanol, and the saponins were concentrated in 90% ethanol solution. The content of saponin part eluted from HP-20 column was 77.62%.

CONCLUSIONThe proposed approach allows convenient and efficient preparation and purification of saponin in Radix Cynanchi Atrati.

Absorption ; Benzaldehydes ; chemistry ; Calibration ; Cynanchum ; chemistry ; Ethanol ; chemistry ; Perchlorates ; chemistry ; Porosity ; Reproducibility of Results ; Resins, Plant ; chemistry ; Saponins ; chemistry ; isolation & purification ; Sensitivity and Specificity

OBJECTIVETo study the chemical features of CPB-4, a heteropolysaccharide obtained from Cynanchum paniculatum.

METHODSugar composition analysis, methylation analysis, partial hydrolysis and carbon-13 nuclear magnetic resonance were used to determine the sugar composition, linkages, main chain, branch chains and branching points.

RESULTCPB-4 is composed of L-arabinose, L-xylose, L-rhamnose and D-galactose in closely molar ratios of 0.8:0.2:0.2:1.0. Its main chain is comprised of 1, 5 linked galactose and side chains are comprised of terminal xylose, terminal arabinose, oligosaccharide of arabinose and oligosaccharide of arabinose, rhamnose and galactose. The branching points are located at C-6 and C-2 of galactose.

CONCLUSIONCPB-4 is a new heteropolysaccharide from C. paniculatum.

Arabinose ; isolation & purification ; Cynanchum ; chemistry ; Methylation ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Polysaccharides ; chemistry ; isolation & purification ; Rhamnose ; isolation & purification ; Xylose ; isolation & purification