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A new isobenzoisofuran(1) has been isolated from the whole plant of Cassia pumila using various chromatographic techniques, including silica gel, Sephadex, MCI-gel resin, and RP-HPLC, and its structure was determined as 9-(2-hydroxyethyl)-2,2-dimethyl-2H-furo[3,4-g]chromen-6(8H)-one. This compound was also evaluated for its antibacterial activity. The results showed that it had prominent antibacterial activity with MIC_(90) value of(45.2±4.2) μg·mL~(-1) for methicillin resistant Staphylococcus aureus(MRSA) strain. This value was closed to that of levofloxacin [with MIC_(90) value(48.5±4.3) μg·mL~(-1)].

The main chemical constituents of naphthopyrone reference extract( NRE) with definite content and relatively fixed chemical composition were analyzed and determined. Ultra-high performance liquid chromatography-LTQ-Orbitrap XL mass spectrometry and high performance liquid chromatography were used to systematically study NRE from the aspects of main chemical components and determination. The results showed that the chemical composition of naphthopyrone reference extract of Cassiae Semen was relatively fixed,and seven naphthalopyranones were identified. Cassiaside B_2,cassiaside C_2,rubrofusarin-6-O-β-D-gentiobioside and cassiaside C were the main chemical constituents of NRE,of which the determination and uncertainty results were( 11. 40+ 0. 26) %,( 11. 68+0. 24) %,( 16. 60+0. 22) %,( 28. 8+0. 48) %,respectively. This study contributed to the accurate evaluation of NRE and the foundation for the application of NRE in the quality control of Cassiae Semen,and provided a new idea for the replacement of single chemical reference substance by the reference extract of traditional Chinese medicine.

References and our previous experiment showed that the contents of glycosides were significantly decreased，while the contents of aglycones were significantly increased after processing of Cassiae Semen.It may be related to its glycosidases or the heating process. In order to investigate the reasons, high performance liquid chromatographic (HPLC) was used to study the effects of these two factors on contents of Cassiae Semen's main chemical components in processing. The results showed that glycoside hydrolases was present in Cassiae Semen and could rapidly hydrolyze glycosides from Cassiae Semen into aglycones in suitable temperature with sufficient water.However，it didn't show effect on contents change of main constituents in the procedure of Cassiae Semen processing.The reason for content decrease of glycosides and content increase of aglycones in processed Cassiae Semen was glycoside bond cracking to produce corresponding aglycone at high temperature.This study further provides basis for further revealing of the processing mechanism of Cassiae Semen., Competing Interests: The authors of this article and the planning committee members and staff have no relevant financial relationships with commercial interests to disclose., (Copyright© by the Chinese Pharmaceutical Association.)

To study the differences and similarities in pharmaceutical characterization and pharmacodynamic characterization between the single decoction and merger decoction of Baihu and Guizhi. The same technology parameters were used to prepare Baihu and Guizhi single decoticon and merger decoction extracts, and then the differences and similarities in pharmaceutical characterization were analyzed based on their HPLC fingerprint, content of index components, and the extraction content. The pharmacodynamic differences and similarities were analyzed by inflammatory model and pain model. There was no significant difference in HPLC chromatographic peak, but the peak area value reflected the difference of quantity to some extent. It was found that the peak value of single Rhizoma anemarrhenae and Cassia twig decoction was less than the peak of their merger decoction, but the peak value of single honey-fried Licorice root decoction was greater than the peak of merger decoction． The contents of neomangiferin, mangiferin and timosaponin B Ⅱ among index components as well as extraction content in merger decoction were higher than those in single decoction． The contents of liquiritin and glycyrrhizic acid as well as extraction content in merger decoction were lower than those in single decoction． There was no significant difference in the content of cinnamicacid and its extraction content between merger decoction and single decoction. According to the efficacy experiment, both of them showed significant anti-inflammatory and analgesic effects. However, the merger decoction showed faster anti-inflammation effect, and longer analgesic effect. It can be concluded that the merger decoction and single decoction of Baihu and Guizhi have the same material basis, and the merger decoction is better for the dissolution of the active ingredients in this recipe, and is more beneficial to the therapeutic effect., Competing Interests: The authors of this article and the planning committee members and staff have no relevant financial relationships with commercial interests to disclose., (Copyright© by the Chinese Pharmaceutical Association.)

A quality control method of Semen Cassiae was established by using naphthopyrone reference extract(NRE). Meanwhile, the feasibility about NRE replacing single component reference in quality control of traditional Chinese medicine was explored. After NRE of Semen Cassiae being prepared by chromatographic separation technology, we determined the three main components, cassiaside B2, rubrofusarin-6-O-β-D-gentiobioside and cassiaside C. In the meantime, an HPLC analytical method, based on the NRE as standard substance, was developed to determinate the contents of three main components in Semen Cassiae. T-test was used for comparison of the determination results of the two methods(single chemical component and NRE as reference substances, respectively), and the T-test result demonstrated that there was no significant difference between the two methods. The results developed scientific basis for the application of NRE of Semen Cassia in the quality control, which could be applied for the quality control of traditional Chinese medicine using reference extract substituting single chemical reference, and provide a new research model for the quality control of Chinese medicine., Competing Interests: The authors of this article and the planning committee members and staff have no relevant financial relationships with commercial interests to disclose., (Copyright© by the Chinese Pharmaceutical Association.)

A new furan-2-carboxylic acid, 5-[3-(hydroxymethyl)-4,5-dimethoxyphenyl]-3-methylfuran-2-carboxylic acid(1),has been isolated from the bark of Cassia alata by using various chromatographic techniques. It displayed cytotoxicity against NB4, A549, SHSY5Y, PC3 and MCF7 cell lines with IC₅₀ values of 2.5, 1.2, 2.2, 3.6 and 1.9 μmol•L⁻¹, respectively., Competing Interests: The authors of this article and the planning committee members and staff have no relevant financial relationships with commercial interests to disclose., (Copyright© by the Chinese Pharmaceutical Association.)

A new isoindole alkaloid (1), has been isolated from the leaves of Cassia siamea by using various chromatographic techniques. Compound 1 is a new compound, determined as 5-(hydroxymethyl)-2-methyl-6-prenylisoindolin-1-one, and it displayed cytotoxicity against NB4, A549, SHSY5Y, PC3 and MCF7 cell lines with IC₅₀ values of 3.2,4.6,2.8,6.4, 2.5 μmol•L⁻¹, respectively., Competing Interests: The authors of this article and the planning committee members and staff have no relevant financial relationships with commercial interests to disclose., (Copyright© by the Chinese Pharmaceutical Association.)

To study the effect of aqueous extract of Cassiae Semen on the activity, mRNA and protein expressions of cytochrome P450(CYP450) system in rat liver microsomes, microsomes of rat liver were prepared after the oral administration with aqueous extract of Cassiae Semen for 14 days. The enzyme activity was quantified by Cocktail method. Meanwhile, the mRNA and protein expressions of CYP1A2, CYP2B1, CYP2C11, CYP2D2, CYP2E1 and CYP3A1 in the livers were detected by RT-PCR and Western blot. The result of this experiment was that aqueous extract of Cassiae Semen obviously induced the enzyme activities of CYP1A2, CYP2B1, CYP2C11, CYP2D2, CYP2E1 and CYP3A1. Low dose of aqueous extract of Cassiae Semen significantly reduced the activity of CYP2D2, but the activity of CYP2D2 was significantly induced by middle dose and high dose of aqueous extract of Cassiae Semen. These subtypes were increased in a dose-dependent manner except for CYP3A1. The mRNA levels of CYP1A2, CYP2C11, CYP2D2 and CYP2E1 were also induced in rats treated with aqueous extract of Cassiae Semen, but with no significant effect in CYP2B1 and CYP3A1 mRNA expressions. The protein levels of CYP2C11 and CYP2E1 were also induced in rats treated with aqueous extract of Cassiae Semen, but with no significant difference. Since the enzyme activity, mRNA and protein expressions of CYP450, particularly CYP2C11and2E1subtypes, were induced or inhibited by aqueous extract of Cassiae Semen to varing degrees, suggesting the potential drug-drug interactions should be concerned., Competing Interests: The authors of this article and the planning committee members and staff have no relevant financial relationships with commercial interests to disclose., (Copyright© by the Chinese Pharmaceutical Association.)

The dry seeds of Cassia obtusifolia were carried by the "ShenZhou 8" satellite and sowed after landing. Based on our pri- or study on SP1, the characteristics of plants growth, physiological index and content of effective components were examined. The results showed that the QC10, QC29 strains matured 5 d earlier compared with control. The plant height, across diameter and ground diameter of QC10, QC29, QC46 strains was superior to the control at whole growth period. The branch number increased ranging from 4 to 11 and the number of pods reached 321, 313,281, respectively, which was dramatically higher than the control (246). The yield of QC10, QC29, QC46 strains increased noticeably from 31.4 to 63.2 g. The 1000-seed-weight of QC10, QC29, QC46 strains was 25.86, 25.88, 24.06 g, while the control was 23.69 g. Compared to the control, the mass fraction of chlorophyll was enhanced 1.098, 1.016, 0.297 mg. There was no significant difference in aurantio-obtusin and chrysophanol content of seeds. Through two years research, three high-yield mutant strains were obtained. This study indicates that spaceflight-induced mutants could provide new germplasm for C. obtusifolia breeding and offers the theoretical basis for further utilization of spaceflight-induced mutation to breed high-quality C. obtusifolia strains.

Cassiae Semen is a common traditional Chinese medicine, and contents of anthraquinones of Cassiae Semen different significantly from area to area. According to Chinese Pharmacopoeia (2010 edition), only contents of aurantio obtusin and chrysophanol were used to evaluate the quality of Cassiae Semen, another data could be added later. Ten batches of Cassiae Semen from different areas were determined, and total anthraquinones, total free anthraquinones and total combined anthraquinones contents were assessed by ultraviolet visible spectrophotometer, contents of aurantio obtusin, rhein, aloe emodin, emodin, chrysophanol and physcion were determined by HPLC. After that, principal components analysis was used to evaluate these data determined previous by dimension reduction analysis. At last, the result suggests that three main components were found out, it shows that content of aloe emodin could be used to evaluate the quality of Cassiae Semen as well as contents of aurantio obtusin and chrysophanol. And Cassiae Semen from Hebei province posseses higher quality than Cassiae Semen from other different areas. All these results can provide a good reference for quality evaluating of Cassiae Semen medicinal materials at a certain extent.

Objective: To investigate the effect of total anthraquinone in Cassiae Semen on lipid peroxidation and peroxisome proliferator activated receptors gamma (PPAR-gamma) expression in liver tissues of rats with alcoholic fatty liver., Method: Referring to literature, it was established animal models of fatty liver feeding with alcohol. Rats were randomized into 6 groups, except the normal group, the other 5 groups of rats had been administered alcohol two times a day for 3 months. Rats were killed at the end of this experiment. It were respectively measured that the contents of ALT, AST, AKP, TG, TC, HDL-C, LDL-C, MDA, SOD, FFA in the serum and TG, TC, MDA, SOD, HL, LPL, FFA in the liver. The left leaf of liver was observed by histopathological staining, the immunohistochemical staining and RT-PCR were used to observe the effects on the expressions of PPAR-gamma mRNA., Result: Compared with the model group, total anthraquinone in Cassiae Semen could remarkably decrease the content of ALT, AST, TC, TG, MDA and increase the content of SOD in the serum of the experimental fatty liver induced by alcohol; remarkably decrease the content of TC, TG, FFA and increase the content of HL, LPL, SOD in the liver of the experimental fatty liver with induced by alcohol. Total anthraquinone in Cassiae Semen group was the similar the model group, but remarkably lighten inflammatory cell intiltration and fibrosis increasing. The RT-PCR and immunohistochemical staining results showed that: compared with the normal group,the model group could remarkably decrease the expression of PPAR-gamma mRNA in the liver (P < 0.01). Compared with the model group, total anthraquinone in Cassiae Semen could remarkably increase the expression of PPAR-gamma mRNA in the liver of the experimental fatty liver (P < 0.01)., Conclusion: The results showed that total anthraquinone in Cassiae Semen has good effects on the treatment of hepatic fat induced by alcohol diet in rats. the possible action mechanism of total anthraquinone in Cassiae Semen possess obvious effect of regulating the disorder of lipid metabolism, ameliorating hepatic function, as well as anti-lipidperoxidation, increasing the expression of PPAR-gamma in hepatic cells of rats.

Objective: To establish a HPLC method for the simultaneous determination of 4 major components in semen cassiae obtusifoline and provide valuable data for quality control of semen cassiae obtusifoline., Method: Kromasil 100-5 C18 column (4.6 mm x 250 mm, 5 microm). Eluted with water: acetonitrile: tetramethylene oxide: glacial acetic acid (100: 23: 5:1). Detecte at 278 nm. The flow rate was 1.0 mL x min(-1)., Result: Four compositions were separated well. Good linearies were obtained within the range of 0.274-1.37 microg, 0.153-0.765 microg, 0.302-1.51 microg and 0.052-0.26 microg for rubrofusarin gentiobioside, casside, aurantio-obtusin-6-O-beta-D- glucopyranoside, casside B respectively. The average recoveries were 100.9% (RSD 1.8%),101.2% (RSD 1.2%), 99.40% (RSD 2.2%), 100.5% (RSD 1.6%)., Conclusion: The method is accurate, reliable and convenient. It can be used to control the quality of semen cassiae obtusifoline and its products.

Objective: To study the chemical constituents of the roasted seeds of Cassia obtusifolia, and to illuminate the change of its effective components before and after being roasted., Method: The compounds were isolated and repeatedly purified by macroporous resin, silica gel column chromatography. Their structures were elucidated by physical and chemical properties and NMR data., Result: Three components were obtained from ethanol extract, and the structures were identified as nor-rubrofusarin-6-O-beta-D-(6'-O-acetyl) -glucopyranoside (1), 1-desmethyl- aurantio-obtusin-2-O-beta-D-glucopyranoside (2), obtusin (3)., Conclusion: Compounds 1 and 2 were isolated from the roasted seeds of C. obtusifolia for the first time, and compound 1 was a new compound.

Objective: To observe the different effects of water soluble extracts from Cassia obtusifolia and processed C. obtusifolia on anti-oxidation and vasodilatation, and to explore the mechanism of eye-clearing effect., Method: Mice were randomly divided into three groups: normal control group, C. obtusifolia group and processed C. obtusifolia group. Ten male and 10 female mice were in each group. mice were given extracts orally 10 g (20 mL) crude drug/kg for one month, control group were given same volume water. Then the eyeball tissue was harvested to prepare homogenate to test SOD, MDA, LDH and NO. Blood was collected from posterior orbital venous plexus to test ET., Result: Both two kinds of extracts could raise the activity of SOD and LDH, decrease the MDA level. Extracts from C. obtusifolia showed higher effects on above mentioned action. Both two kinds of extracts increased the ratio value of NO/ET significantly, and extracts from processed C. obtusifolia showed higher activity., Conclusion: Both C. obtusifolia and processed C. obtusifolia extracts show obvious anti-oxidation and vasodilatation effects. These results may support the eye-clearing activity of C. obtusifolia.

Objective: To establish methods of RP-HPLC respectively for content-determination of two types of constituents in the Cassia obtusifolia respectively, and tp determine the constituents between the raw and roasted seeds of C. obtusifolia in order to distinguish the difference of those seeds., Method: HPLC systems consisted of Alltima C18 (4.6 mm x 150 mm, 5 microm), column temperature at 30 degrees C, flow rate of 1.0 mL x min(-1). Two different mobile phases and detection wavelengths: I , ACN-THF-0.1% H3 PO4 (17 : 3:80), 278 nm; II , MeOH-0.1% H3PO4 (79:21), 254 nm. Analyzed and compared the content of the two types of constituents between the raw and roasted seeds of C. obtusifolia., Result: The calibrate curves of 5 constituents were linear (r > 0.999 7). The precision and repeatability were perfect (RSD < 1.6%, 1.8%). The samples were stabile during 24 h., Conclusion: The study provide a better universal reference for evaluating and controlling the quality of C. obtusifolia pieces.

Objective: To isolate and identify chemical constituents of Cassia alata., Method: Compounds were separated and purified by column chromatography with silica gel and Sephadex LH-20, elucidated by spectroscopic methods including 1D and 2D NMR, IR and MS techniques., Result: Twelve compounds were isolated from C. alata, which were identified as chrysoeriol (1), kaempferol (2), quercetin (3), 5,7,4'-trihydroflavanone (4), kaempferol-3-O-beta-D-glucopyranoside (5), kaempferol-3-O-beta-D-glucopyranosyl-(1-->6)-beta-D-glucopyranoside (6), 17-hydrotetratriacontane (7), n-dotriacontanol (8), n-triacontanol (9), palmitic acid ceryl ester (10), stearic acid (11), palmitic acid (12)., Conclusion: Compounds 3-12 were isolated from C. alata for the first time.

Objective: To establish HPLC fingerprint for the identification of Cassia obtusifolia and compare chemical constituents of the raw and roasted seeds of C. obtusifolia., Method: Chromatographic fingerprint of C. obtusifolia was investigated by RP-HPLC and the gradient elution mode was applied to chromatographic separation. Data were analyzed by fingerprint similarity evaluation software to compare the similarity of the samples for identifying the main chromatographic peaks furthermore., Result: Having compared the HPLC fingerprint of the raw and roasted seeds of C. obtusifolia were compared preliminarily, and 12 main chromatographic peaks were identified., Conclusion: HPLC method can be used in quality control and identification of the raw and roasted seeds of C. obtusifolia.

Objective: To investigate the chemical constituents of the roasted seeds of Cassia obtusifolia to illuminate the change of its effective components before and after roasted., Method: Compounds were separated by silica gel chromatography, and their structures were evaluated by spectral analysis and chemical evidence., Result: Seven compounds were isolated from the ethanol extract. Their structures were identified as chrysophanol (1), physcion (2), 8-methoxylchrysophanol (3), beta-sitosterol (4), emodin (5), obtusin (6) and obtusifolin-2-O-beta-D-(6'-O-acetyl) glucopyranside (7)., Conclusion: Compounds 1-7 were isolated from the roasted seeds of C. obtusifolia for the first time, and compound 7 was a new compound.

Objective: To determine the content of 7 anthraquinones in Semen Cassiae., Method: A HPLC method was developed, with Inertsil ODS-3 column, acetonitrile and 0.1% H3PO4 solution as mobile phases in gradient elution. The detection wavelength wasset at 278 nm, and the flow rate was 0.8 mL x min(-1)., Result: Recoveries of all 7 anthraquinones were between 95%-105%. The content of the anthraquinones in crude drug produced in different habitation were different., Conclusion: The method is convenient and accurate, which provides the foundation for the research of Semen Cassiae.

Objective: To establish a method for determining the content of rubrofusarin gentiobioside in Cassia obtusifolia., Method: The HPLC with Diamonsil C18 (4.6 mm x 250 mm, 5 microm) column was used, acetonitrile-THF-1% acetic acid (18: 3:79) was used as a mobile phase, with flow rate of 1 mL x min(-1), column temperature at 40 degrees 2 and detection wavelength at 278 nm., Result: A good linearity was obtained from 0.1-0.5 microg with r = 0.999 9 for rubrofusarin gentiobioside. The average recovery was 101.1%, and RSD was 2.23% (n = 5)., Conclusion: The method was proved to be simple, rapid, sensitive, precise, reliable and repeatable. It can be applied to the quality control of Semen Cassia.

Objective: To investigate actions of protein and anthraquinone glucosides from Semen Cassia on learning and memory capacity and related substances of senile mice induced by D-galactose., Method: The subacute senile mouse models induced by injection of D-galactose subcutaneously were used., Result: Protein and anthraquinone glucosides from Semen Cassia could improve the learning and memory capacity of senile mice. Protein and anthraquinone glucosides from Semen Cassia not only inhibited the increase of malondialdehyde (MDA) in cerebrum of senile mice, but also enhanced the level of superoxide dismutase (SOD) in cerebrum and lessened the lipofuscin (LF) in liver tissue of senile mice. Protein from Semen Cassia could reduce the content of monoamine oxidase (MAO) in cerebrum of senile mice., Conclusion: Protein and anthraquinone glucosides from Semen Cassia could improve the learning and memory capacity of senile mice and delay aging.

Objective: To study the chemical constituents in hairy roots of Cassia obtusifolia., Method: The hairy roots of C. obtusifolia were induced with Agrobacterium rhizogenes LBA9402 from cotyledons and cultured in MSO liquid medium. The compounds were isolated by silica gel, polyamide and Sephadex LH-20 column chromatography, and the structures were elucidated by employing chemical and spectral methods, Result: Eight compounds were isolated from the ethyl acetate fraction of 95% EtOH extract of the transformed roots of C. obtusifolia. They are betulinic acid, chrysophanol, physcion, stigmasterol, 1-hydroxy-7-methoxy-3-methyl-anthraquinone, 8-O-methylchrysophanol, 1-O-methylchrysophanol and aloe-emodin, and aloe-emodin was isolated from the hairy roots of C. obtusifolia for the first time., Conclusion: The hairy roots of C. obtusifolia have the ability to synthesize the similar chemical constituents as the original plants.

Objective: To analysie the anthraquinones absorbed into serum by different animals (or human beings) after oral administration of Semen Cassiae., Method: Anthraquinones in serum and urine of rats and urine of healthy men after taking Semen Cassiae orally were detected with HPLC., Result: Only some of the anthraquinones were absorbed into serum. There were differences in absorption and metabolism of anthraquinones between rats and men and some new anthraquinones were produced in the process., Conclusion: Anthraquinones absorbed into serum by the experimental animals or men should become target for researching into active compounds of anthraquinones in Semen Cassiae.

Objective: To identify Cassia seeds of six species by capillary electrophoresis., Method: The water-soluble extracts of Cassia seeds of six species were analyzed by capillary electrophoresis. The running buffer was 0.1 mol.L-1 borate, 0.1 mol.L-1 SDS, pH 8.5. The separation voltage was 25 kV., Result: Four common peaks could be found in the electropherograms of six species Cassia seeds, and the characteristic peaks could also be observed., Conclusion: Fingerprints of the six species of Cassia seeds show significant differences, which can be used for their identification.

Objective: To study the effect of protein and anthraquinone glucosides from cassia seed on serum lipid of hyperlipidemia rats., Method: The rat hyperlipidemia model was set up by ig lipid emulsion. The effects of the protein 0.25 and 1 mg.kg-1. d-1, the anthraquinone glucosides 5 and 20 mg.kg-1.d-1, and the protein 0.25 mg.kg-1.d-1 plus the anthracene glucosides 5 mg.kg-1.d-1 on total cholesterol (TC), triglyceride (TG), low density lipoprotein (LDL-C) and high density lipoprotein (HDL-C) in serum of the rats were determined., Result: The protein 1 mg.kg-1.d-1, the anthraquinone glucosides 20 mg.kg-1.d-1 reduced the raised TC, TG, LDL-C of hyperlipidemia rats (P < 0.05). The above indexes could also be reduced by the protein 0.25 mg.kg-1.d-1 plus the anthraquinone glucosides 5 mg.kg-1.d-1(P < 0.05, P < 0.01)., Conclusion: Protein and anthraquinone glucosides from cassia seed can lower TC, TG, LDL-C of hyperlipidemia rats.

Objective: To study the influence of decocting time on the amount of anthraquinone diffused from the cassia seeds., Method: The amount of anthraquinone in the decoctions made in different lengths of decocting time was determined by spectrophotometry (for intact seeds and powdered seeds)., Result: Fast nonlinear diffusion district, slow linear diffusion district and ceasing diffusion district were shown on the light absorption-decocting time curves, and appropriate smashing of the cassia seed facilitated the diffusion of anthraquinone., Conclusion: The best decocting time for anthraquinone to diffuse from the cassia seeds should be the ending time of the fast nonlinear diffusion district, while the second decoction should not be given up.

A HPLC method for the determination of chrysophanol in Semen Cassiae was investigated. The result showed that this method was simple, specific and accurate. The recovery was 99.0% and relative standard deviation was 1.8%. The method is useful for the quality control of this medicinal meaterial.