OBJECTIVE:To establish a method for the contents determination of phospholipid and γ-linolenic acid in Com-pound linolenic acid soft capsule. METHODS:HPLC was conducted to determine the content of soybean lecithin;the column was Agilent TC-C18 with mobile phase of methanol-water(84:16,V/V)at a flow rate of 1.0 ml/min,the detection wavelength was 219 nm,the column temperature was 30 ℃,and the injection volume was 10 μl.Gas chromatography was conducted to determine the content of γ-linolenic acid in the preparation;the column was DB WAX,injector temperature was 210 ℃ by temperature pro-grammed,carrier gas was helium at a flow rate of 1 ml/min by split injection(split ratio of 1:30),and the injection volume was 0.02μl. RESULTS:The linear range was 4.64-16.24μg/ml for phospholipid(r=0.999 6)and 0.093 44-0.327 04μg/ml forγ-linole-nic acid methy ester(r=0.999 6);RSDs of precision,stability and reproducibility tests were lower than 3%;recoveries were 97.44%-99.36%(RSD=0.93%,n=6) and 97.22%-99.07%(RSD=1.01%,n=6),respectively. CONCLUSIONS:The method is simple,stable with good separation,and can be use for the contents determination of phospholipid and γ-linolenic acid in Com-pound linolenic acid soft capsule.

OBJECTIVE:To establish the HPLC fingerprint for Anemone raddeana. METHODS:HPLC was performed on the column of Phemomenex Gemini C18 with mobile phase of 0.1%phosphoric acid-acetonitrile(gradient elution)at a flow rate of 1 ml/min,the detection wavelength was 206 nm,the column temperature was 30℃,and the injection volume was 20μl. With the refer-ence of raddeanin A,13 batches of A. raddeana were analyzed,chromatographic fingerprint similarity evaluation system software was conducted for similarity analysis,and SPSS 13.0 was conducted for cluster analysis. RESULTS:There were 11 common peaks in the 13 batches of A. raddeana with similarity of higher than 0.90. According to the verification,the fingerprint and control fin-gerprint shows good consistency. The drugs in Huadian,Jiaohe, Tiangang,Shulan,Tonghua and Fusong of Jilin and Shangzhi of Heilongjiang were regarded as category 1,and in Harbin,Yabuli town and Yimianpo of Heilongjiang,Qingyuan of Liaoning,Ji-nan of Shandong were category 2. CONCLUSIONS:The established fingerprint can provide reference for the identification and quality evaluation of A. raddeana.

OBJECTIVE:To establish a method for the content determination of rosin acid in Rheumatoid arthritis tablet. METH-ODS:HPLC was performed on the column of ZORBAX SB-C18 with mobile phase of acetonitrile-0.1% formic acid(82∶18,V/V) at flow rate of 1.0 ml/min ,detection wavelength was 241 nm ,column temperature was 30 ℃ and volume injection was 10 μl. MS/MS column was ZORBAX SB-C18 with mobile phase of acetonitrile-0.1% formic acid(80∶20,V/V)at flow rate of 0.2 ml/min;column temperature was 30 ℃;volume injection was 0.5 μl. Ionization mode was ESI+,atomization gas pressure was 25 psi,gas flow as 8.0 L/min,capillary voltage was 4 000 V,capillary outlet voltage was 120 V,precursor ion was 303 m/z,scan range was 50-500 m/z and the gas temperature was 350 ℃. RESULTS:The linear range of rosin acid was 2.5-100.0 μg/ml. RSDs of preci-sion,stability and reproducibility tests were lower than 2.0%,recoveries was 96.75%-98.11%(RSD=0.53%,n=6). CONCLU-SIONS:The method is simple,accurate and reproducible,and suitable for the content determination of rosin acid in Rheumatoid arthritis tablet.

OBJECTIVE To establish the grade s tandard for Panax quinquefoli um and to evaluate the quality of different grades of medicinal materials. METHODS Totally 24 batches of P. quinquefolium were used as test samples. Pearson correlation analysis method was used to analyze the correlation between qualitative analysis indicators （taproot length ，taproot diameter and weight of single root ）and internal component indicators （ethanol-soluble extract ，and the contents of ginsenoside Rg 1，ginsenoside Re ， ginsenoside Rb 1，ginsenoside Rc ，ginsenoside Rb 2，ginsenoside Rd ，pseudo-ginsenoside F 11）. Combined with chemometrics methods，the reference indexes for the classification of P. quinquefolium were selected ，and the classification standards were formulated. HPLC-ELSD fingerprints of 24 batches of P. quinquefolium were established and their similarity evaluation was also performed. The chromatographic peaks were identified by comparison with the reference substance ，and then the quality of different grades of P. quinquefolium was evaluated by cluster analysis. RESULTS After screening ，taproot diameter ，the weight of single root and the content of ginsenoside Rd were taken as the reference indexes for the classification of P. quinquefolium . According to above 3 indexes，P. quinquefolium were divided into 3 grades：special grade ，first grade and second grade. According to the center value of K-means clustering ，the total score of special-grade medicinal materials was more than 135.40，that of first-grade medicinal materials was 61.82-135.40，and that of second-grade medicinal materials was less than 61.82. In the HPLC-ELSD fingerprints of 24 batches of P. quinquefolium ，25 common peaks were confirmed ，and 7 characteristic peaks were identified. The similarity of the chromatograms of P. quinquefolium of special grade ，first grade and second grade with fingerprints ranged 0.980-0.989，0.962-0.968，0.940-0.949，respectively. The results of cluster analysis showed that different grades of P. quinquefolium could be identified significantly. CONCLUSIONS The grade standard and HPLC-ELSD fingerprints of P. quinquefolium are established，which can be applied for exclusive identification of P. quinquefolium ，and provide reference for its quality control and grade classification.

ObjectiveTo explore the possible mechanism of Danggui Sini Granules （当归四逆颗粒） in treatment of coronary heart disease with cold congealing and blood stasis syndrome. MethodsFifty SD rats were randomly divided into a blank group， a model group， a Danshen Pill （丹参滴丸） group， and a low- and high-dosage Danggui Sini Granules group， with 10 rats in each group. Except for the blank group， the rat model of coronary heart disease with cold congealing and blood stasis syndrome was established by repeated cold stimulation at low temperature combined with intraperitoneal injection of posterior pituitary hormone in all other groups. In the 6th week of modelling， 0.073 g·kg^-1·d^-1 of compound Danshen Pill was given to the Danshen Pill group， 20.2 and 40.4 g·kg^-1·d^-1 of Danggui Sini Granules were given to the low- and high-dose Danggui Sini Granules groups， respectively， and 0.2 ml/10 g of sterile water was given to the blank group and the model group， all for 2 consecutive weeks. The general conditions of the rats were recorded， and the body mass was compared weekly. At the end of the intervention， electrocardiogram， blood rheological indexes， including whole blood low-cut viscosity， whole blood high-cut viscosity， erythrocyte aggregation index， and cardiac index were detected to evaluate the effect of the medication， and HE staining was used to observe the myocardial histopathological changes， TUNEL staining to detect the apoptotic situation of cardiomyocytes， and ELISA to detect the serum thyroid-stimulating hormone （TSH）， free triiodothyronine （FT₃）， free tetraiodothyronine （FT₄） levels and thromboxane B₂ （TXB₂）， 6-keto-prostaglandin F1α （6-Keto-PGF_1α）， endothelin 1 （ET-1）， and nitric oxide （NO） levels. ResultsCompared with the blank group at the same time point， the body mass of rats in the remaining groups decreased at all time points （P＜0.05 or P＜0.01）； compared with the model group at the same time point， the body mass of rats in high-dose Danggui Sini Granules group and Danshen Pill group increased at the 6th and 7th weeks （P＜0.05 or P＜0.01）. In the blank group， the S-T segment of the electrocardiogram was close to the isoelectric line， the myocardial structure was regular， the fibres were closely arranged， and the nuclei of the cells were intact and neatly aligned； in the model group， the S-T segment of the electrocardiogram significantly elevated， and the arrangement of the myocardial fibres was obviously disordered， with myocardial cells appearing to be swollen， necrotic， and infiltrated by inflammatory cells， and the apoptosis-positive cells of the cardiac muscle cells obviously increased； in each of the medication groups， the electrocardiogram had a lowered S-T segment， and myocardial fibres were aligned， myocyte structure and morphology were improved， inflammatory cells reduced， and the number of apoptosis-positive cells significantly reduced. Compared with the blank group， the cardiac index， whole blood high cut viscosity， whole blood low cut viscosity， erythrocyte aggregation index increased in the model group， and the serum levels of TSH， FT₃， FT₄， NO， 6-Keto-PGF_1α significantly reduced， and the levels of ET-1 and TXB₂ significantly increased （P＜0.05 or P＜0.01）. Compared with the model group， the levels of cardiac index， whole blood high cut viscosity， whole blood low cut viscosity， erythrocyte aggregation index， ET-1 and TXB₂ significantly decreased and the levels of NO and 6-Keto-PGF_1α significantly increased in each medication group， and the serum levels of TSH， FT₃， and FT₄ elevated in the high-dose Danggui Sini Granules group and Danshen Pill group （P＜0.05 or P＜0.01）. The serum TSH level of rats in the low-dose Danggui Sini Granules group was lower than that in Danshen Pill group and the high-dose Danggui Sini Granules group （P＜0.01）. ConclusionDanggui Sini Granules have the effect of alleviating myocardial injury in coronary heart disease with cold congealing and blood stasis syndrome， and its mechanism may be related to reducing myocardial cell apoptosis， improving energy metabolism， and regulating the level of vasoactive factors.

ObjectiveTo observe the effects of Xuefu Zhuyu capsules （XFZY） on blood lipid levels and aortic plaques in the mouse model of atherosclerosis （AS） induced by a high-fat diet by regulating the silencing regulatory factor 2-like protein 3 （Sirt3）/exchange protein directly activated by cAMP 1 （EPAC1） signaling pathway and explore the mechanism of XFZY in ameliorating AS. MethodMice were assigned into normal， model， blank， rosuvastatin （0.05 g·kg^-1·d^-1）， and low-， medium-， and high-dose （0.3， 0.6， 1.2 g·kg^-1·d^-1， respectively） XFZY groups. The normal group consisted of normal C57BL/6J mice， while the other groups consisted of ApoE^-/- C57BL/6J mice. The normal group and blank group were fed routinely， and the rest groups were fed with a high-fat diet for 24 consecutive weeks for the modeling of AS. The drug intervention groups were administrated with corresponding drugs by gavage， and model group and blank group with an equal volume of deionized water for 6 consecutive weeks. The small animal B-ultrasound was used to evaluate the mouse heart function and aortic plaque condition. A fully automated biochemical analyzer was used to measure the levels of blood lipids such as total cholesterol （CHOL）， triglycerides （TG）， high-density lipoprotein cholesterol （HDL-C）， low-density lipoprotein cholesterol （LDL-C）， and extremely low-density lipoprotein （VLDL） in mice. Oil red O staining was employed to observe lipid deposition in the aorta. Hematoxylin-eosin staining and Masson staining were employed to observe the pathological changes and collagen deposition in mouse blood vessels. Transmission electron microscopy was employed to observe the mitochondrial damage in mouse aorta. The levels of adrenocorticotropic hormone （ACTH）， adenosine triphosphate （ATP）， and nicotinic choline receptor α1 （CHRNα1）， and total superoxide dismutase （T-SOD） were measured by enzyme-linked immunosorbent assay （ELISA）. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and Western blot were performed to determine the mRNA and protein levels， respectively， of Sirt3， EPAC1， Caspase-3， B-cell lymphoma-2 （Bcl-2）， and Bcl-2-associated X protein （Bax） in the mouse aorta and heart. ResultMultiple AS plaques were observed in the aortic arch， indicating that the model was successfully established. Compared with the model group， the XFZY groups showed reduced and narrowed plaques. Compared with the normal group， the model group showed elevated CHOL level （P<0.01）. Compared with the model group， rosuvastatin and low-dose XFZY lowered the CHOL and TG levels （P<0.01）. Compared with the normal group， the model group presented a large number of protruding red lipid plaques on the aortic wall and increased percentage of AS plaque area to total tissue area （P<0.01）. Compared with the model group， low-dose XFZY reduced the plaque load （P<0.05）. Compared with the model group， XFZY at different doses reduced the lipid plaques and collagen deposition. Compared with the normal group， the model group showed decreased or disappeared mitochondrial cristae and presented severe damage of the membrane structure in endothelial cells. The mitochondria of endothelial cells in each treatment group approached the normal structure， with mitochondrial cristae faintly visible. Compared with the normal group， the model group showcased reduced myocardial mitochondrial ATP activity （P<0.01）， which were rescored in the drug intervention groups （P<0.01）. Compared with the normal group， the modeling inhibited the expression of Sirt3 （P<0.01） and promoted the expression of EPAC1 （P<0.01）. Compared with the model group， low-dose XFZY increased the Sirt3 content （P<0.01） and medium-dose XFZY increased the EPAC1 content （P<0.01）， which indicated that XFZY treatment upregulated the mRNA and protein levels of Sirt3 and downregulated the mRNA and protein levels of EPAC1. ConclusionXFZY can alleviate the aortic lipid deposition， reduce the AS plaque area， improve the mitochondrial morphology and functions in endothelial cells， increase the ATP activity， upregulate the expression of Sirt3， and downregulate the expression of EPAC1 in AS mice by regulating mitochondrial energy metabolism via the Sirt3/EPAC1 signaling pathway.

ObjectiveTo analyze and determine the differential components of freeze-dried and sun-dried Panacis Quinquefolii Radix（PQR）， and to compare the differences in their pro-angiogenic activities. MethodFingerprints of freeze-dried and sun-dried PQR were established based on ultra performance liquid chromatography（UPLC）， and chemometrics methods such as principal component analysis（PCA） and orthogonal partial least squares-discriminant analysis（OPLS-DA） were combined to determine the differential saponin composition of the two decoction pieces， and six representative saponins were selected and their contents in freeze-dried and sun-dried PQR were determined by UPLC. Transgenic zebrafish line Tg（fli1a∶EGFP） embryos fertilized for 24 h were selected， and different doses of 70% methanol extracts of freeze-dried and sun-dried PQR（10， 30 mg·L^-1） were used to intervene in normal zebrafish and in a zebrafish model of intersegmental vascular（ISV） injury induced by vascular endothelial growth factor（VEGF） receptor tyrosine kinase inhibitor Ⅱ（PTK787）， then the development of subintestinal vein（SIV） and ISV of zebrafish was observed， SIV diameter， mean number of crossings and mean number of germinations were determined， and the ISV vascular index was calculated， in order to compare the pro-angiogenic activities of the two decoction pieces. ResultThe similarity of the fingerprints of freeze-dried and sun-dried PQR decoction pieces was>0.950， and 17 common peaks were identified， of which 6 common peaks were designated as peak 6（ginsenoside Rg₁）， peak 7（ginsenoside Re）， peak 8（ginsenoside Rb₁）， peak 11（ginsenoside Rc）， peak 13（ginsenoside Rb₂）， and peak 16（ginsenoside Rd）， respectively. A total of 11 differential saponin components were screened by PCA and OPLS-DA， indicating that there were some differences in the contents of the components in the two decoction pieces. The results of determination showed that the contents of ginsenosides Rg₁， Re， Rb₁ and Rb₂ in freeze-dried PQR were higher than those in sun-dried PQR， while the contents of ginsenosides Rc and Rd were lower than those in sun-dried PQR（P<0.05， P<0.01）. In the study of the pro-angiogenic effect on normal zebrafish embryos， compared with the blank group， and the SIV vessel diameter， mean germination rate and mean crossover rate were significantly higher in the high-dose groups of freeze-dried and sun-dried PQR（P<0.01）， and the vessel diameter， mean numbers of crossings and germinations in the freeze-dried PQR group were higher than those of the sun-dried PQR group（P<0.05）. In the study of the pro-angiogenic effect on zebrafish embryos with ISV injury， the development of ISV in the model group was significantly inhibited when compared with the blank group， compared with the model group， different dose groups of freeze-dried and sun-dried PQR could promote the growth and sprouting of ISV， and the number of normal blood vessels in the freeze-dried PQR group was significantly higher than that in the sun-dried PQR group at the same dosage（P<0.05）. ConclusionFreeze-drying can effectively avoid the loss and secondary transformation of ginsenosides in PQR， and its angiogenic activity is better than that of sun-dried PQR， which can provide a reference for the production and development of high-quality PQR decoction pieces.

ObjectiveTo analyze the quantity-quality transfer of standard decoction of Ginseng Radix et Rhizoma（GRR） decoction pieces produced by fresh and traditional cutting， and to provide reference for quality control and application development of the decoction pieces produced by fresh cutting. MethodTen batches of representative GRR decoction pieces produced by fresh and traditional cutting and their standard decoctions were prepared by standard process， and high performance liquid chromatography（HPLC） fingerprint of the standard decoction was established and performed on an Agilent EC-C₁₈ column（4.6 mm×150 mm， 2.7 μm） with acetonitrile（A）-0.1% phosphoric acid aqueous solution（B） as the mobile phase for gradient elution（0-23 min， 18%-21%A； 23-35 min， 21%-28%A； 35-80 min， 28%-32%A）， and the detection wavelength was 203 nm. Then similarity evaluation， principal component analysis（PCA） and partial least squares-discriminant analysis（PLS-DA） of fingerprint of the standard decoction were performed to screen the differential components with variable importance in the projection（VIP） value>1. Quantitative analysis was carried out on the screened known differential components， and combined with the indicators of the dry extract rate and the transfer rate， to explore the differences in the quantity-quality transfer between the standard decoction of GRR decoction pieces produced by fresh and traditional cutting. ResultThe fingerprint similarity of the standard decoction of GRR decoction pieces produced by fresh and traditional cutting was more than 0.950， and 18 common peaks were identified， including 9 identified common peaks. The results of PCA and PLS-DA showed that there were some differences in the contents of index components between the two standard decoctions. The contents of ginsenoside Rg₁， Re and Ro in GRR decoction pieces produced by fresh cutting were higher than those in traditional decoction pieces， while the contents of ginsenoside Rb₁， Rc ， Rb₂ and Rd were lower than those in traditional decoction pieces. The contents of ginsenoside Rg₁， Re， Rb₁ and Ro in the standard decoction of GRR decoction pieces produced by fresh cutting were higher than those in the standard decoction of traditional decoction pieces， while the contents of ginsenoside Rc ， Rb₂ and Rd were comparable between the two standard decoctions. Compared with the standard decoction of the traditional decoction pieces， the average transfer rates of ginsenoside Rg₁， Rb₁， Rc， Rb₂ and dry extract rate of the standard decoction of GRR decoction pieces produced by fresh cutting were significantly increased（P<0.05）， and the average transfer rate of ginsenoside Re and Rd also increased， but the difference was not statistically significant. ConclusionThe dry extract rate， content and transfer rate of index components of standard decoction of GRR decoction pieces produced by fresh cutting are better than those of the standard decoction of traditional decoction pieces， which can provides data support for the subsequent clinical application of fresh cutting products.