[Research advances of biosynthesis, in vivo analysis and pharmacokinetics of chemical constituents in Artemisiae Annuae Herba].

Artemisiae Annuae Herba is a traditional Chinese medicine for clearing deficiency and heat. It is the only natural source of artemisinin, which is a specific antimalarial drug, and has been widely concerned all over the world. In addition to artemisinin, Artemisiae Annuae Herba also contains many sesquiterpenes, coumarins, flavonoids, volatile oils, polysaccharides and other chemical components, which show antipyretic, anti-inflammatory, antiviral microorganisms, anti-asthma, anti-oxidation, anti-tumor and other pharmacological activities. In addition to their own pharmacological activities, some components could enhance the antimalarial activity of artemisinin through different mechanisms at absorption and metabolism in vivo. In order to understand the pharmacokinetic characte-ristics of the chemical constituents contained in Artemisiae Annuae Herba and provide reference for the full development and clinical utilization of Artemisiae Annuae Herba resources in China, this present paper systematically collated the modern research literatures, and summarized the biosynthesis, in vivo analysis and pharmacokinetics of the chemical constituents in Artemisiae Annuae Herba.

[Construction of artesunate nanoparticles modified by hyaluronic acid and cell-penetrating peptides and its inhibitory effect on cancer cells in vitro].

Hyaluronic acid (HA) and cell-penetrating peptide (CPP) R6H4-SA modified artesunate nanostructured lipid carrier (HA-R6H4-NLC/ART) for anti-tumor therapy was prepared. The physicochemical properties and in vitro drug release of HA-R6H4-NLC/ART were evaluated, and the uptake and cytotoxicity of liver cancer HepG2 cells were studied. The results showed that HA-R6H4-NLC/ART was spherical like in appearance, and the average particle size was about 160 nm. In vitro release experiments showed that the drug delivery system had sustained release characteristics. Cell results showed that, in slightly acidic environment, pH sensitive CPP R6H4-SA mediated cellular uptake of nanoparticles was significantly higher than that of non-sensitive peptide R8-SA. Meanwhile, HA-R6H4-NLC/ART had a targeting effect on HepG2 cells, and the HA receptor saturation experiment showed that the endocytosis of HA-R6H4-NLC/ART was mediated by the HA receptor on the cell surface. As compared with the unmodified or R6H4-SA single modified group, HA and R6H4-SA co-modified HA-R6H4-NLC/ART significantly improved the cell uptake and had a stronger anti-tumor effect under the conditions of the slightly acid environment and hyaluronidase degradation. The above results showed that hyaluronic acid and CPP R6H4-SA co-modified artesunate nanostructured lipid carrier, which can effectively identify and penetrate the tumor cell membrane into the cell, is a potentially efficient targeting delivery system for anti-tumor drugs.

[History of processing of Sojae Semen Praeparatum].

Sojae Semen Praeparatum (SSP) is commonly used as a type of dietetic Chinese herb. By collecting and analyzing ancient and recent literatures, a textual criticism was conducted on the historical evolution of the processing of SSP. Fermented soybean was recorded in Shijing, and relevant rational processing was described in Qimin Yaoshu. In the early time, fermented soybean included the type of "salty" and "light". After the Ming Dynasty, "light" fermented soybean or SSP was recognized as a better medicinal matter than salty fermented soybean, and the fermentation processing was recorded more clearly. In modern time, many characteristic methods for processing SSP have been developed. Today, the processing of SSP is mainly based on the Chinese Pharmacopoeia, which records soybean as a main ingredient and Artemisiae Annuae Herba, Mori Folium as excipients.

Two new guaiane-type sesquiterpenoids from Valeriana hardwickii and their cytotoxicity.

Two new guaiane-type sesquiterpenoids, named 4α,5α-epoxy-8ß-hydroxy-1α-hydro-α-guaiene (1) and 4α,5α-epoxy-1-hydroxy-α-guaiene (2), were isolated from the whole plants of Valeriana hardwickii. Their structures were elucidated on the basis of spectroscopic analysis. Compounds 1 and 2 showed weak cytotoxicity against the lung adenocarcinoma (A549) and hepatoma (Bel7402) cell lines with IC50 values of 9.2 and 8.5 µM, respectively.

Effects of arteannuin B, arteannuic acid and scopoletin on pharmacokinetics of artemisinin in mice.

OBJECTIVE: To explore the effects of arteannuin B, arteannuic acid and scopoletin on the pharmacokinetics of artemisinin in mice. METHODS: Artemisinin and a combination of artemisinin, arteannuin B, arteannuic acid and scopoletin were administered together to mice via oral administration. Blood samples were collected at different time intervals and pretreated by liquid-liquid extraction. The contents of four compounds in mouse plasma were determined by a validated HPLC-MS/MS method. RESULTS: Compared to single artemisinin group, the Cmax values from the combination group rose from 947 ng/mL to 1254 ng/mL. AUC(0-t) (2371 h ng/mL) was significantly higher than that from single artemisinin group (747 h ng/mL). The peak time lag and the CL values reduced at a proportion of 66%. CONCLUTIONS: Arteannuin B, arteannuic acid and scopoletin can markedly affect the pharmacokinetics of artemisinin.

[Chemical constituents from whole plants of Valeriana hardwickii].

Chemical investigation of the whole plants of Valeriana hardwickii has led to the isolation of 11 flavones and 2 monoterpe- noids by using various chromatographic techniques including column chromatography on silica gel and Sephadex LH-20, preparative TLC, and preparative HPLC. Their structures were identified by spectroscopic data analysis as syzalterin (1), 6-methylapigenin (2), 5-hydroxy-7,4'-dimethoxyflavone (3), genkwanin (4), acacetin (5), apigenin (6), quercetin (7), tricin (8), (-)-farrerol (9), sosakuranetin (10), 5,3',4'-trihydroxy-7-methoxyflavanone (11), (-)-bornyl ferulate ( 12) , and (-)-bornyl caffeate ( 13). All compounds were isolated from this plant for the first time, while compounds 1, 9-13 were obtained from this genus for the first time.

[Scientific connotation of processing Bombyx Batryticatus under high temperature].

The aim of this study was to elucidate the scientific connotation of Bombyx Batryticatus processing with wheat bran under high temperature. The contents of soluble protein extracted from Bombyx Batryticatus and its processed products and the limited content of AFT in Bombyx Batryticatus and the processed one were compared. The concentration of protein was measured with the Bradford methods and the difference of protein between Bombyx Batryticatus and its processed products was compared by SDS-PAGE analysis. Aflatoxin B1, B2, G1, and G2 were determined by reversed-phase HPLC. The results showed that the soluble protein content of Bombyx Batryticatus and its processed products were (47.065 +/- 0.249), (29.756 +/- 1.961) mg x g(-1), correspondingly. Analysis of protein gel electrophoresis showed that there were no significant differences between the crude and processed one in protein varieties. 6 bands were detected: 31.90, 26.80, 18.71, 15.00, 10.18, 8.929 kDa. Below 10 kDa, the color of bands of the processed one was deeper than the crude one, which demonstrate that macromolecular protein was degradated into micromolecule. The content of AFG1, AFB1, AFG2, AFB2 were 0.382, 0.207, 0.223, 0.073 g x kg(-1), not exceeded 5 microg x kg(-1) while the processed one was not detected. Through processing with wheat bran under high temperature, the content of soluble protein in Bombyx Batryticatus decreased, the processing purpose for alleviating drug property was achieved. Meanwhile, the limited content of aflatoxins were reduced or cleared by processing procedure or absorbed by processing auxillary material, adding the safety of the traditional Chinese Medicine. In conclusion, as a traditional processing method, bran frying Bombyx Batryticatus was scientific and reasonable.

[Simultaneous quantitation of artemisinin, arteannuin B, artemisic acid, and scopoletin in mice plasma by HPLC-MS].

The objective of this study is to develop a sensitive and reliable high-performance liquid chromatography mass spectrometry (LC-MS) method for simultaneous determination of artemisinin, arteannuin B, artemisic acid, and scopoletin, and study the pharmacokinetics of the four constituents in mouse serum after oral administration of the four components to mice. The analytical column used was Agilent Zorbax SB-C18 (2.1 mm x 150 mm, 5 mm). The mobile phase was acetonitrile: 0.5% acetic acid (60: 40) and the flow rate was 0.3 mL x min(-1). The temperature of the column was 40.0 degrees C. In this condition, we established an analysis method to simultaneously determine the four components. A sensitive and specific liquid chromatography-mass spectrometric (LC-MS) method was developed and validated for the determination of artemisin in derivatives in mice plasma. The method we established has a linear range of 5-3 000 µg x L(-1) with a good sensitivity and specificity for all of the four components. This method is simple, rapid, accurate and suitable for the determination of the content of the four compounds.

[Artemisinin resistance in Plasmodium falciparum: global status and basic research].

Artemisinin-resistant Plasmodium falciparum has been identified by WHO in the Greater Mekong subregion. While there is no report on artemisinin resistance in Africa and South America by now, related surveillance measures have been taken place. The genes related artemisinin-resistance has been identified and the molecular markers will be used for large-scale surveillance efforts to contain artemisinin resistance. The emergence and spread of artemisinin resistance worldwide is a present danger and needs more attention. This article reviews the progress of artemisininresistance malaria parasites and artemisinin-based combination therapies.

[Triterpenoids from Stauntonia obovatifoliola Hayata subsp. intermedia stems].

In the current study, a total of nineteen triterpenoids (1-19) from 60% EtOH extracts of Stauntonia obovatifoliola Hayata subsp. intermedia stems were separated and purified by solvent extraction and chromatographic methods including silica gel, ODS as well as preparative HPLC. According to the results of chemical reactions and spectral data, compounds were identified as: lupeol (1), betulinonic acid (2), betulinic acid (3), 3-epi-betulinic acid (4), quinatic acid (5), 24-O-acetyl quinatic acid (6), 3-O-α- L-arabinopyranosyl-30-nor-hederagenin-28-O-α-L-rhamnopyranosyl-(1 --> 4) -ß-D-glucopyranosyl-(1 --> 6) -ß-D-glucopyranosyl ester (7), Stauntoside A (8), kalopanax saponin A (9), kalopanax saponin J (10), Kizuta saponin K10 (11), 3-O-α-L-rhamnopyranosyl (1--> 2) -α-L-arabinopyranosyl-hederagenin-28-O-ß-D-xylopyranosyl-(1 --> 6) -ß-D-glucopyranosyl ester (12), kalopanax saponin B (13), 3-O-α-L-rhamnopyranosyl-(1 --> 2) -α-L-arabinopyranosyl-hederagenin-28-O-ß-D-glucopyranosyl-(1 --> 6) -ß-D-glucopyranosyl ester (14), sieboldianoside A (15), septemoside A (16), kalopanax saponin K (17), septemloside I (18), and 3-O-α-L-arabinopyranosyl (1 --> 2)-ß-D-glucuronopyranosyl- hederagenin (19). Among them, compounds 4, 6, 10, 12, 14, and 16-19 were isolated from the Stauntonia genus for the first time, and compound 6 was a new natural product.

[Study on intervention effect of Danggui Shaoyao San on rats with cirrhotic ascites].

OBJECTIVE: To investigate the intervention effect of Danggui Shaoyao San on rats with cirrhotic ascites, and discuss the effect of arginine vasopressin (AVP) on cirrhotic ascites. METHOD: Male SD rats were randomly divided into the control group, the model group, Danggui Shaoyao San low, middle and high dose groups. The cirrhotic ascites rat model was established by CCl4 combined with phenobarbital. Their urines were collected at 24 h to observe urine excretion of each group. Filter papers were used to determine the amount of ascites. The levels of serum alanine aminotransferasa (ALT) , aspartate aminotransferase (AST) were detected by the automatic biochemistry analyzer. Plasma prothrombin time (PT) was evaluated by the blood coagulation analyzer. The concentration of AVP in plasma was detected by enzyme-linked immunosorbent assay (ELISA). Pathological changes in livers were observed by HE staining. RESULT: Compared with the model group, the Danggui Shaoyao San group showed significant improvement in live indexes, with notable decrease in serum ALT and AST and the time of PT, improvement in liver pathological changes. Simultaneously, the amount of ascites decreased to varying degrees, with notable increase in urine in 24 h and decrease in AVP concentration in plasma. CONCLUSION: Danggui Shaoyao San can notably improve liver functions of rats with cirrhotic ascites, reduce the generation of ascites and delay the progress of liver pathological changes. Its mechanism may be related to AVP.

A new beta-naphthalenecarboxylic acid biglycoside from Chirita longgangensis var. hongyao.

To investigate the chemical constituents in the stems of Chirita longgangensis var. hongyao, methanol extract of the stems was subjected to column chromatography with various chromatographic techniques. One new beta-naphthalenecarboxylic acid biglycoside, 1, 4-dihydroxy-2-naphthalenecarboxylic acid methyl ester-4-O-alpha-L-rhamnopyranosyl-(1-->6)-beta-D-glucopyranoside (1) was isolated, along with two known compounds: isotaxiresinol 4-O-methyl ether (2) and (R)-7-hydroxy-alpha-dunnione (3). Compound 2 was first obtained from Chirita genus and compound 3 was isolated from this plant for the first time. All structures were elucidated on the basis of spectral and chemical evidence, and the NMR spectroscopic data of compound 2 was published for the first time.

Simple HPLC evaluation of lipoamidase activity in tissue using a newly synthesized fluorescent substrate, dansyl-α-lipoyllysine.

α-Lipoic acid (LA) is a naturally occurring disulfide-containing compound used as an antioxidant supplement which also has been used as a medicine for diabetic neuropathy in Europe. Physiologically LA acts as a coenzyme of mitochondrial multienzyme complex in its protein bound form but it is not yet clear how the externally administrated LA is incorporated into other proteins in the same protein-bound form or why the bound form is active as an antioxidant. The binding and cleavage of LA to or from the protein is mediated by lipoamidase and thus determines LA distribution in tissues. We have developed a simple sensitive assay for lipoamidase using a fluorescent substrate, dansyl-α-lipoyllysine (DLL). Lipoamidase in tissues cleaves the amide bond between LA and the ε-amino-lysine moiety to release dansylated lysine (DL). A HPLC comparison of the fluorescence intensity between DLL and DL was used to quantify the enzyme activity. The hydrolytic reaction did not occur when the tissue was heat-treated before incubation with DLL and was inhibited by free LA, especially by the R-enantiomer of LA (physiologically active form). N(ε)-Acetyl-L-lysine did not compete with DLL in the cleavage reaction. The method was applied for the determination of lipoamidase activity levels in various rat tissues. It was revealed the spleen had the highest activity followed by the kidney, heart, lung and liver. The activity in the brain was below the detection limit of the assay.

[Determination of artemisinin, arteannuin B and artemisinic acid in Herba Artemisiae Annuae by HPLC-UV-ELSD].

To establish an HPLC-UV-ELSD method for the determination of the content of artemisinin, arteannuin B and artemisinic acid in Herba Artemisiae Annuae. The analytical column was Nucleodur RP-C18 (250 mm x 4.6 mm, 5 microm ID). The mobile phase was acetonirile-0.1% acetic acid (50: 50) and the flow rate was 1.0 mL x min(-1) with a UV detector for artemisinin, the detection wavelength at 209 nm, and the evaporative light-scattering detector (ELSD) for arteannuin B and artemisinic acid, the drift tube temperature: 50 degrees C, the nitrogen flow rate 30 psi and the gain was 50. The resolution of artemisinin, arteannuin B and artemisinic acid was good. The linear calibration curves were obtained over the range of 0.52 - 2.6 microg for artemisinin (r = 0.999 4, n = 5), 0.022 - 4.4 microg for artemisinin B (r = 0.999 9, n = 5) and 0.203 - 8.12 microg for artemisinic acid (r = 0.999 8, n = 5), separately. The mean recoveries of the three compounds were 99.45%, 102.37% and 101.10% with RSD of 2.3%, 1.7% and 0.79%, respectively. This method is simple, rapid, accurate and suitable for the determination of the content of the three compounds in the herbs.

[Studies on the chemical constituents from stem of Chirita longgangensis var. Hongyao].

OBJECTIVE: To isolate and elucidate the constituents from stem of Chirita longgangensis var. hongyao. METHOD: The constituents were extracted with methanol and isolated by chromatography on silica gel, Sephadex LH-20 and ODS. The structures were determined by NMR and MS spectral analysis. RESULT: Seven compounds were identified as 2-hydroxy-7-methyl-9, 10-anthraquinone (1), 2-methyl-9, 10-anthraquinone (tectoquinone) (2), ursolic acid (3), vanillic acid (4), beta-sitosteryl-D-glucoside-6'-palmitate (5), beta-sitosterol (6), daucosterol (7), respectively. CONCLUSION: Compounds 1, 2, 3 and 5 were isolated for the first time from the family Gesneriaceae, compounds 4 and 6 were isolated for the first time from the genus Chirita, and compound 7 was isolated for the frist time from Chirita longgangensis var.

[Phenylethanoid glycosides from stem of Chirita longgangensis var. hongyao].

OBJECTIVE: To isolate and elucidate the constituents from stem of Chirita longgangensis var. hongyao. METHOD: The constituents were extracted with methanol and isolated by chromatography on silica gel, Sephadex LH-20 and ODS. The structures were determined by NMR and MS spectral analysis. RESULT: Five phenylethanoid glycosides were identified as 3, 4-dihydroxyphenyl alcohol-beta-D-glucopyranoside (1), 3, 4-dihydroxyphenyl alcohol-6-O-caffeoyl--D-glucopyranoside (calceolarioside B) (2), 3, 4-dihydroxyphenyl alcohol-beta-D-glucopyranosyl-(1 --> 3)-6-O-caffeoyl-beta-D-glucopyranoside (plantainoside D) (3), 3, 4-dihydroxyphenyl alcohol-beta-D-glucopyranosyl-(1 --> 3)-4-O-caffeoyl-beta-D-glucopyranoside (plantamajoside) (4), 3, 4-dihydroxyphenyl alcohol-beta-D-glucopyranosyl-(1 --> 3)-6-O-feruloyl-beta-D-glucopyranoside (scroside E) (5), respectively. CONCLUSION: Compounds 3 and 5 were isolated for the first time from the family Gesneriaceae, and compounds 1, 2 and 4 were isolated for the first time from the genus Chirita.

[Studies on chemical constituents in rhizome of Matteuccia struthiopteris].

OBJECTIVE: To study the chemical constituents in rhizome of Matteuccia struthiopteris. METHOD: The compounds were isolated by normal phase silica gel chromatography. The structures were identified by physical and spectral data. RESULT: Six compounds were isolated and identified as woodwardic acid (1), ergost-6,22-diene-3beta,5alpha,8alpha-triol (2), apigenin (3), riboflavin (4), 4-O-beta-D-glucopyranosyl-p-coumaric acid (5), 4-O-beta-D-glucopyranosylcaffeic acid (6). CONCLUSION: All the compounds were obtained from this plant for the first time.