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Separation and regeneration of protoplast from Phellinus igniarius.

China Journal of Chinese Materia Medica.2007;32(21):2232-2235.

OBJECTIVETo study the conditions on separation and regeneration of protoplast from Phellinus igniarius.

METHODThe effects of enzymolysis conditions of P. igniarius mycelia on yield of protoplast and culturing conditons on regeneration ratio of protoplast were investigated.

RESULTWhen the 8 days-old mycelia was hydrolysed by 1.5% of lywallzyme adding to driselase of 0. 5% and at 30 degrees C for 3 h and enzymolysis was stablized by sucrose as a stablisher of osmotic pressure, higher yield of P. igniarius protoplast was obtained. If 10 days-old mycelia was used as raw material of enzymolysis and manntol was selected as stablisher of osmotic pressure of enzymolysis, higher regeneration ratio of P. igniarius protoplast also would be obtained in following regeneration step at same time keeping higher yield. For the regeneration processing, it was beneficial for the regeneration of P. igniarius protoplast that PDA plusing mulberry ramulus was used as the culture medium of regeneration and manntol was selected as the osmotic pressure establisher of regeneration culture medium.

CONCLUSIONThe method and conditions to keep both higher yield and regeneration ratio of P. igniarius protoplast were obtained.

Culture Media ; pharmacology ; Fungal Proteins ; pharmacology ; Glucan Endo-1,3-beta-D-Glucosidase ; pharmacology ; Glycoside Hydrolases ; pharmacology ; Mannitol ; pharmacology ; Multienzyme Complexes ; pharmacology ; Osmotic Pressure ; Peptide Hydrolases ; pharmacology ; Polyporaceae ; drug effects ; physiology ; Protoplasts ; drug effects ; physiology ; Regeneration ; drug effects ; Sucrose ; pharmacology ; Temperature

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Cloning and bioinformatics analysis of P450 cDNA in Artemisia annua.

Jian-Qiang KONG ; Wei WANG ; Xiao-Dong ZHENG ; Ping ZHU ; Ke-Di CHENG

China Journal of Chinese Materia Medica.2007;32(21):2227-2231.

OBJECTIVECloning and bioinformatics analysis of P450 cDNA in Artemisia annua.

METHODA P450 cDNA gene was cloned from A. annua by RT-PCR. The bioinformatics analysis of the P450 gene was performed.

RESULTThe complete ORF of this P450 cDNA is 1 464 bp and encodes 488 aa. The sequence was reported to GenBank and coded as DQ667171. Bioinformatics analysis of the P450 cDNA showed it encoded an A-type P450 protein with 54. 992 kDa, it's isoelectric point was 8.83 and the possibility of export to mitochondria was 0.893 2.

CONCLUSIONThe comparable analysis of the P450 with CYP71AV1 revealed that the two proteins probably performed the same function because of the similar character.

Amino Acid Sequence ; Artemisia annua ; enzymology ; genetics ; Base Sequence ; Cloning, Molecular ; Cytochrome P-450 Enzyme System ; classification ; genetics ; DNA, Complementary ; chemistry ; genetics ; Molecular Sequence Data ; Open Reading Frames ; genetics ; Phylogeny ; Plant Proteins ; genetics ; Plants, Medicinal ; enzymology ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Analysis, DNA ; Sequence Homology, Amino Acid

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Effects of density on seedling growth and glycyrrhizinic acid content in Glycyrrhiza uralensis.

Zhi-Rong SUN ; Ming-Pu ZHAI ; Wen-Quan WANG ; Yu-Rong LI

China Journal of Chinese Materia Medica.2007;32(21):2222-2281.

OBJECTIVETo study the principle of interaction among individuals and populations in different Glycyrrhiza uralensis growth density.

METHODThe phenological characteristics, growth status, accumulation and distribution of biomass and glycyrrhizinic acid content of G. uralensis in different growth density were studied.

RESULTDensity had effect on phenological characteristics and growth status of G. uralensis seedling. The fast growing period of plant was advanced with the increase of the density, while the duration of fast growing period was decreased. Individual biomass decreased with the increase of the density, while population biomass increased. Biomass proportion of root, stem and leaf in individual were 65.02%, 19.55% and 15.43% respectively. Their relative standard deviations were 0.57, 0.49 and 0.76 respectively. Glycyrrhizinic acid content was 0.52% -0.59% under low density and 0.29% under high density.

CONCLUSIONThe result showed that the density G. uralensis had a negative correlation with the shooting time of offshoot and underground stem, leaflet number, growth of plant height and diameter at ground, root head diameter, lateral root number, individual root and branch-leaf number, individual biomass and glycyrrhizinic acid content and a positive correlation with the population biomass.

Agriculture ; methods ; Biomass ; Ecosystem ; Glycyrrhiza uralensis ; growth & development ; metabolism ; Glycyrrhizic Acid ; analysis ; metabolism ; Plant Leaves ; growth & development ; metabolism ; Plant Roots ; growth & development ; metabolism ; Plant Stems ; growth & development ; metabolism ; Plants, Medicinal ; growth & development ; metabolism ; Seedlings ; growth & development ; metabolism

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Systematic studies on quality of main species of Herba epimedii.

Li-Kuan PEI ; Wen-Hua HUANG ; Tian-Gu HE ; Bao-Lin GUO

China Journal of Chinese Materia Medica.2007;32(21):2217-2222.

OBJECTIVETo achieve the quality control of Herba Epimedii, and to evaluate the quality of medical materials of the 8 official species including the 5 species recorded in 'Chinese Pharmacopoeia' including Epimedium brevicornum, E. sagittatum, E. pubescens, E. koreanum and E. wushanense, and the 3 other species (E. acuminatum, E. myrianthum, E. leptorrhizum) recorded in 'Guizhou Quality Criteria for Traditional Chinese Medical Material and Nationality Medical Material', and E. pseudowushanuse (new species) which is used as E. wushanense for a long time.

METHODThe contents of icariin and total flavonoids of 102 samples of 9 officinal species of Herba Epimedii were determined by HPLC and UV, respectively.

RESULT AND CONCLUSIONThe contents of icariin in about 30% of the samples of the 5 species recorded in 'Chinese Pharmacopoeia' were lower than 0.5%, which is acceptable quality recorded in 'Chinese Pharmacopoeia'. Refering the literatures, we suggested the total contents of epimedin A, B, C and icariin (epi-medium multi-glycosides, ABCI) should be established as a new standard instead of the content of icariin. The content of total flavonoids, not less than 5.0%, and ABCI, not less than 1.3%, could be used to evaluate the quality of the above medical materials efficiently.

Drugs, Chinese Herbal ; analysis ; standards ; Epimedium ; chemistry ; classification ; Flavonoids ; analysis ; Flavonols ; analysis ; Pharmacognosy ; Plants, Medicinal ; chemistry ; classification ; Quality Control ; Reproducibility of Results ; Seasons ; Species Specificity

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Effect of Rehmannia glutinosa water extraction on insulin resistance and gene expression of resistin in type 2 diabetes mellitus rats.

Xiu-Fang LV ; Qing-Yu MENG ; Xin-Min GUO

China Journal of Chinese Materia Medica.2007;32(20):2182-2184.

OBJECTIVETo investigate the effect of Rehmannia glutinosa water extraction (RGL) on adipose metabolic disorder and gene expression of resistin in type 2 diabetes mellitus rats.

METHODThe wistar rats model of 2-DM were induced by high calorie feeding and small dose injection of STZ. Rats were randomly divided into diabetic model group, diabetic model treated with RGL (2.4 g x kg(-1) x d(-1)), RGL (1.2 g x kg(-1) x d(-1)), RGL (0. 6 g x kg(-1) x d(-1)) and normal control group. The levels of FPG, FINS, TG, HDL, LDL, CH and IR were measured, and the mRNA expression of resistin was determined by RT-PCR, the protein expression measured by SDS-PAGD at the end of 8 weeks.

RESULTThe gene expression of resistin in RGL group were lower than that of diabetic model (P < 0.01). The levels of FPG, FINS, TG, LDL, CH, IR in RGL group were lower than that of diabetic model (P < 0.05), and HDL were higher (P < 0.05). CONCLUDE: RGL can improve insulin resistance in the experimental 2-DM rats, can effectively ameliorate adipose metabolic disturbance and decline IR and FINS by increasing the gene expression of resistin.

Animals ; Blood Glucose ; metabolism ; Cholesterol ; blood ; Diabetes Mellitus, Experimental ; blood ; genetics ; pathology ; Diabetes Mellitus, Type 2 ; blood ; genetics ; pathology ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Electrophoresis, Polyacrylamide Gel ; Female ; Gene Expression Regulation ; drug effects ; Hypoglycemic Agents ; isolation & purification ; pharmacology ; Insulin ; blood ; Insulin Resistance ; Male ; Plants, Medicinal ; chemistry ; RNA, Messenger ; biosynthesis ; genetics ; metabolism ; Random Allocation ; Rats ; Rats, Wistar ; Rehmannia ; chemistry ; Resistin ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Triglycerides ; blood

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Effect of Danxiongfang and its components on experimental liver injury models induced by CCl4 in mice.

Xiao-Rong LI ; Li-Juan WANG ; Yu-Hang LI ; Yan-Xia XU ; Yi CHEN ; Jin-Xiu RUAN ; Ming XUE

China Journal of Chinese Materia Medica.2007;32(20):2177-2181.

OBJECTIVEThe study investigates the protective effect on liver of Danxionfang and its components.

METHODMice are injected with CCl4 to establish liver injured model. ALT, AST, serum albumin, globulin in serum and SOD, MDA in liver and liver histological changes were measured to confirm the ability of protecting liver of Danxiongfang.

RESULTThe results show Danxiongfang can inhibit obviously the abnormal increase of ALT, AST in serum and MDA in liver, enhance SOD activity in liver, total protein, albumin, globulin in serum, and decrease liver pathological changes, which suggests Danxiongfang can protect injured liver induced by CCl4.

CONCLUSIONDanxiongfang showed powerful protective effect against liver damage induced by CCl4.

Alanine Transaminase ; blood ; Animals ; Aspartate Aminotransferases ; blood ; Carbon Tetrachloride ; Chemical and Drug Induced Liver Injury ; Drug Combinations ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Female ; Ligusticum ; chemistry ; Liver ; drug effects ; metabolism ; pathology ; Liver Diseases ; blood ; prevention & control ; Male ; Malondialdehyde ; metabolism ; Mice ; Mice, Inbred ICR ; Phytotherapy ; Plants, Medicinal ; chemistry ; Protective Agents ; isolation & purification ; pharmacology ; Salvia miltiorrhiza ; chemistry ; Superoxide Dismutase ; metabolism

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Effects of CASI on myocardial ischemia-reperfusion arrhythmia in rats.

Li WANG ; Xiao-Feng YU ; Shao-Chun QU ; Hua-Li XU ; Da-Yuan SUI

China Journal of Chinese Materia Medica.2007;32(20):2174-2177.

OBJECTIVETo observe the protective effect of compound acanthopanax senticosus injection (CASI) on myocardial ischemia-reperfusion arrhythmia in rats.

METHODThe myocardial ischemia-reperfusion model was induced by 30 min coronary occulusion and 60 min reperfusion in openchest anesthetized rats. The changes of arrhythmia with electrocardiogram lead II, the activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px), the contents of malondialdehyde (MDA) and Ca2+ in myocardium were determined.

RESULTIn rats treated by CASI (in a dosage of 25, 50 and 100 mg x kg(-1) femoral vein infusion at 30 min after coronary occulusion), the incidence of myocardial ischemia-reperfusion ventricular arrhythmias, for instance the ventricular tachycardia (VT) and ventricular fibrillation (Vf), was effectively prevented, the appearing time of arrhythmia was delayed and the duration of arrhythmia was shortened, while the elevated ST segment lowered as well. At the same time, the contents of myocardial Ca2+ and MDA were decreased significantly as well as the activities of myocardial SOD and GSH-Px increased markedly.

CONCLUSIONCASI is of protective effect on myocardial ischemia-reperfusion arrhythmia, which may be related to scavenging the oxygen free radicals and Ca2+ overload formed during reperfusion.

Animals ; Anti-Arrhythmia Agents ; isolation & purification ; pharmacology ; Arrhythmias, Cardiac ; drug therapy ; etiology ; physiopathology ; Calcium ; metabolism ; Drug Combinations ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Electrocardiography ; Eleutherococcus ; chemistry ; Female ; Ginsenosides ; isolation & purification ; pharmacology ; Glutathione Peroxidase ; metabolism ; Male ; Malondialdehyde ; metabolism ; Myocardial Reperfusion Injury ; complications ; Myocardium ; metabolism ; pathology ; Panax ; chemistry ; Phytotherapy ; Plants, Medicinal ; chemistry ; Rats ; Rats, Wistar ; Saponins ; isolation & purification ; pharmacology ; Superoxide Dismutase ; metabolism

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Effect of Astragalus polysaccharide on pancreatic cell mass in type 1 diabetic mice.

Ru-Jiang LI ; Shu-Dong QIU ; Hong-Xia CHEN ; Hong TIAN ; Guo-Qiang LIU

China Journal of Chinese Materia Medica.2007;32(20):2169-2173.

OBJECTIVETo study the effect of Astragalus polysaccharide (APS) on pancreatic beta cell mass in type 1 diabetic mice.

METHODDiabetic mice induced by multiple low dose streptozotocin (MLD-STZ) were administered either APS (100, 200, 400 mg x kg(-1) body weight) or saline intraperitoneally daily, and sacrificed after 15 or 30 days of treatment. Streptavidin-peroxidase immunohistochemical method with counterstain was performed to determine the effect of APS on insulitis. Indirect double immunofluorescence for Insulin/Ki67 (counterstained by Hoechst33258) and Insulin/Cleaved caspase-3 was used to evaluate pancreatic cell (besides beta cell) proliferation, beta cell neogenesis, beta cell apoptosis and beta cell mass. Semi-quantitative RT-PCR was utilized to characterize pancreatic regenerating protein 1 mRNA levels, and ELISA method was performed to measure the levels of cytokine IFN-gamma and IL-4 secreted by splenocytes.

RESULTAttenuated insulitis, upregulated beta cell mass, increased number of neogenetic pancreas islets, decreased number of apoptosis beta cells and downregulation of Th1/Th2 cytokine ratio were significantly time-and dose-dependent on APS treatment, when compared to saline controls. However, no significant differences of the number of pancreatic proliferative cells or replicative cells and pancreatic regenerating protein 1 mRNA levels were demonstrated between APS (APS100, APS200 and APS400) and saline vehicle group on day 15 and 30 with APS treatment.

CONCLUSIONAPS can upregulate pancreatic beta cell mass in type 1 diabetic mice, strongly associated with improved autoimmunity.

Animals ; Apoptosis ; drug effects ; Astragalus membranaceus ; chemistry ; Carrier Proteins ; metabolism ; Diabetes Mellitus, Experimental ; chemically induced ; metabolism ; pathology ; Diabetes Mellitus, Type 1 ; chemically induced ; metabolism ; pathology ; Enzyme-Linked Immunosorbent Assay ; Insulin-Secreting Cells ; drug effects ; metabolism ; pathology ; Interferon-gamma ; metabolism ; Interleukin-4 ; metabolism ; Islets of Langerhans ; drug effects ; metabolism ; pathology ; Lithostathine ; biosynthesis ; genetics ; Male ; Mice ; Mice, Inbred C57BL ; Plants, Medicinal ; chemistry ; Polysaccharides ; isolation & purification ; pharmacology ; RNA, Messenger ; biosynthesis ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Streptozocin ; Transcription Factors

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Effect of total saponins of Rubus parviflolius (TSRP) on change of hydrated amount and blood-brain barrier in rats during focal cerebral ischemic/reperfusion.

Ji-Sheng WANG ; Zong-Yin QIU ; Hui-Zhi LI ; Yong-Peng XIA ; Cheng-Lin ZHOU

China Journal of Chinese Materia Medica.2007;32(20):2166-2169.

OBJECTIVETo explore the effects of total saponins of Rubus parviflolius (TSRP) on brain edema and blood brain barrier in rats.

METHODThe model of local cerebral ischemia was established in rats by reversible inserting a nylon thread into the anterior cerebral artery through the internal carotid artery brain hydrated amount and content change of Evan' s blue (EB) in cortex subjected to 2h middle cererbral artery occlusion (MACO) followed by 6 h, 24 h, 48 h, 72 h reperfusion and effect of TSRP. penetrability of blood brain-barrier (BBB) the index includes brain hydrated amount and penetrability of blood brain-barrier BBB.

RESULTCom- pared with I/R group. Both brain hydrated amount and the EB content decreased significantly in TSRP groups on the 6 h, 24 h, 48 h, 72 h of reperfusion after 2 hour of cerebral ischemia induced by MACO model.

CONCLUSIONTSRP could decrease brain hydrated amount and markedly lower permeability of blood-brain barrier subjected to 2 h MACO followed by 24 h reperfusion, and this may be a mechanism of TSRP alleviating brain edema during I/R.

Animals ; Blood-Brain Barrier ; drug effects ; Brain Edema ; drug therapy ; etiology ; pathology ; Brain Ischemia ; complications ; Infarction, Middle Cerebral Artery ; complications ; Male ; Phytotherapy ; Plants, Medicinal ; chemistry ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; complications ; Rosaceae ; chemistry ; Saponins ; isolation & purification ; pharmacology

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Experimental studies of effects of Wujiwan extracts in different compatibilities on motility of isolated colon in guinea pig.

Ya-Jie WANG ; Yu DONG ; Xiao-Xin ZHU

China Journal of Chinese Materia Medica.2007;32(20):2161-2165.

OBJECTIVETo investigate the effects of Wujiwan extracts in two different compatibility proportions (named the No. 1 and No. 2 respectively) on motility of isolated colon in guinea pig, and compare the therapeutic effects with each other.

METHODTo observe the function of the No. 1 and No. 2 with different concentrations on colon contraction induced by acetylcholine (Ach) and inhibition induced by adrenalin (Ad) respectively, and calculate the EC, with the method of Bliss.

RESULTThe No. 1 and No. 2 both could significantly inhibit the contraction of colon, and in 15 minutes, there was significant time-dependence and concentration-dependence, and EC50 of the No. 1 was lower than that of No. 2. However, but when the colonic was inhibited by Ad, at the time of 9 min and 15 min, the No. 1 at the level of 300 mg x L(-1), and at the time of 15 min, the No. 2 at the level of 30 mg x L(-1), could both excite the colon significantly (P < 0. 05).

CONCLUSIONOur experiments indicate that the No. 1 and No. 2 both can significantly inhibit the colon contraction in guinea pig, and the No. 1 is superior to the No. 2 to some extent. And at the same time, we also can conclude that the No. 1 and No. 2 have exciting effects on inhibitory colon. Meanwhile, the No. 1 and No. 2 each have its own advantages based on the common therapeutic effects.

Acetylcholine ; pharmacology ; Animals ; Colon ; drug effects ; physiology ; Coptis ; chemistry ; Dose-Response Relationship, Drug ; Drug Combinations ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; pharmacology ; Epinephrine ; pharmacology ; Evodia ; chemistry ; Female ; Gastrointestinal Motility ; drug effects ; Guinea Pigs ; In Vitro Techniques ; Male ; Muscle Contraction ; drug effects ; Paeonia ; chemistry ; Plants, Medicinal ; chemistry

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