OBJECTIVE： To establish the method for the content determination of gentiopicrin and loganic acid in Gentiana scabra， and to investigate the correlation of their contents with appearance traits and quality gradation criterion. METHODS： HPLC method was adopted. The determination was performed on Ascentis Express C18 column with mobile phase consisted of 0.1％ phosphoric acid-acetonitrile （gradient elution） at the flow rate of 0.4 mL/min. The column temperature was 30 ℃， and detection wavelength was set at 240 nm. The sample size was 1 μL. Taking the length， number and diameter of fibrous roots as indexes， the appearance and morphological characteristics of G. scabra were studied. The relationship of gentiopicrin and loganic acid content with the appearance property of medicinal material was analyzed by SPSS 21.0 software. k-mean clustering analysis was carried out by using SPSS 21.0 software， and gradation standard for G. scabra was established preliminarily. RESULTS： The linear range of gentiopicrin and loganic acid were 0.5-3.0 μg/mL （r＝0.999 9） and 0.05-0.50 μg/mL （r＝0.999 9）. The limit of quantification of gentiopicrin and loganic acid were 0.295， 0.289 μg/mL； the detection limit were 0.082， 0.081 μg/mL； RSDs of precision， stability， repeatability tests were all lower than 2％； the recovery rates were 97.56％- 102.23％ （RSD＝1.56％， n＝6） and 97.58％-102.67％ （RSD＝1.86％， n＝6）. Correlation results showed that there was a significant positive correlation of the length of G. scabra， the number of roots， root diameter， with the contents of gentiopicrin and loganic acid. The order of affecting content was the number of roots ＞length ＞root diameter. k-means clustering analysis showed that 54 batches of G. scabra was divided into two categories； S4-S6，S13，S17-S23，S25，S28，S31-S34 were clustered into a category； S1-S3， S7-S12， S14-S16， S24， S26，S27，S29， S30，S35-S54 were clustered into the other category. The results of gradation showed that 54 batches of G. scabra could be divided into two grades， and the results were consistent with the cluster analysis. CONCLUSIONS： Established method is simple and stable， and can be used for simultaneous determination of gentiopicrin and loganic acid in G. scabra. The more fibrous roots， the longer the length， the thicker the root， the higher the content of gentiopicroside and loganic acid， the better the quality of G. scabra.

OBJECTIVE：To investigate t he effects of different compatibility ratio of Gardenia jasminoides to fermented soybean on the content of genistein and total flavonoids ，and to investigate the compatibility regularity of Zhizichi decoction. METHODS：The decoction method was used to prepare the mixed decoction with different compatibility ratio of G. jasminoides to fermented soybean （2∶1，1∶1，1∶2，1∶4，m/m，the same hereinafter ）. UPLC-MS/MS method was used to determine the content of genistein in Zhizichi decoction with different compatibility ratio and corresponding fermented soybean single decoction. UV method was used to determine the content of total flavonoids in Zhizichi decoction with different compatibility ratio and corresponding gardenia single decoction and fermented soybean single decoction. RESULTS ：The established method had good linearity ， precision，repeatability，stability and accuracy. Compared with single decoction ，the content of genistein in the mixed decoction with different compatibility ratio of G. jasminoides to fermented soybean （2∶1，1∶1，1∶2，1∶4）was decreased to different extents ， while the content of total flavonoids was increased to different extents. With the increase of fermented soybean ，the content of genistein in the decoction increased at first and then decreased. When the compatibility ratios of G. jasminoides to fermented soybean were 1 ∶ 1 and 1 ∶ 2，the content of genistein in the decoction was the highest （all 0.071 μg/mL）. With the increase of fermented soybean ，the content of total flavonoids in the decoction did not change regularly ；when the ratio of G. jasminoides to fermented soybean was 1 ∶ 1，the content of total flavonoids in the decoction was the highest （1.861 μg/mL）. CONCLUSIONS ： When the compatibility ratio of G. jasminoides to fermented soybean was 1 ∶ 1，the content of flavonoids in the decoction is the highest.

OBJECTIVE：To simultaneo usly determine the contents of atractylenolide Ⅱ ，β-eudesmol，atractyloxin and atractylone in Atractylodes chinensis ，and to evaluate the quality of A. chinensis with different growth years combined with color difference principle. METHODS ：HPLC method was adopted. The determination performed on Agilent Eclipse XDB-C 18 column with mobile phase consisted of acetonitrile- 0.2％ phosphoric acid （gradient elution ）at the flow rate of 1.0 mL/min；the detection wavelengths were set as 208 nm（atractylenolide Ⅱ，β-eudesmol），340 nm（atractyloxin）and 220 nm（atractylone）；the sample size was 15 μ L. Using atractyloxin as reference，QAMS was adopted to establish relative correction factors （RCFs） of atractylenolideⅡ，β-eudesmol and atractylone ；the content of each component in A. chinensis with different growth years were calculated. The contents of above 4 components were determined by external standard method and then compared with the results of QAMS. The color difference values of A. chinensis powder were measured based on color difference principle. The correlation analysis of above 4 components content with color was carried out by Pearson correlation analysis. RESULTS ：The separation degree of atractylenolide Ⅱ，β-eudesmol，atractyloxin and atractylone in A. chinensis was higher than 1.5. The linear range were 1.01-10.10，3.30-33.00，4.40-44.00，5.34-53.40 μg/mL，respectively. RSDs of precision ，reproducibility and stability tests were all lower than 2％，while the average recovery rates were 101.34％-104.67％（RSD＜1.5％，n＝6）. Using atractyloxin as reference ， RCFs of atractylenolide Ⅱ，β-eudesmol and atractylone were 3.896 7，5.928 2，9.727 9，with RSD of 0.35％，2.89％，0.36％ （n＝6），respectively. Relative deviation of 3 components （except for atractyloxin ） in 24 batches of A. chinensis ranged 0.03％-1.45％ between QAMS and external standard method ，which indicated that the results of two methods were consistent ，and the content of each component increased with the increase of growth years. Atractylenolide Ⅱ，β-eudesmol，atractyloxin and atractylone in A. chinensis had significant negative correlation with its color shade （L*），total color difference （E*ab）（P＜0.01）， and significant positive correlation with color red-green direction （a*）， color yellow-blue direction （b*）（P＜0.01）. CONCLUSIONS：The established HPLC-QAMS method can be used for the determination of atractylenolide Ⅱ，β-eudesmol， atractyloxin and atractylone in A. chinensis . The longer the growth period is ，the higher each component content is. The color of A. chinensis is closely related to the content of each component ，and the content of effective components is higher in A. chinensis with dark yellowish brown color.

ObjectiveTo investigate the relationship of the polymorphism of endothelial nitric oxide synthase (eNOS), the 27-bp variable number of tandem repeats (VNTR) in intron 4 with essential hypertension (EH) of the northern Han nationality in China.MethodsGenotypes, the level of plasma nitric oxide metabolites (NOx) and the activity of nitric oxide synthase (NOS) of 207 EH subjects and 231 healthy subjects were measured by polymerase chain-reaction (PCR).ResultsThe frequencies of ecNOS4a/a,ecNOS4b/a, and ecNOS4 b/b in the healthy group were 0.43%, 13.42% and 86.15% respectively. The frequency of the b allele was 92.86%, and the frequency of the a allele was 7.14%. While the frequencies of ecNOS4 a/a, ecNOS4 a/b,and ecNOS4 b/b in the EH group were 0.49%, 19.32% and 80.19% respectively. The frequency of the a allele in EH group (n=42, 10.15%) was significantly higher than that in the healthy group (n=33, 7.14%)(P<0.05). The plasma NOx level of the EH group was 70.04±14.68 mol/L, and significantly lower than that 84.09±27.27 mol/L in the healthy group (P<0.05). Similarly, both the plasma TNOS and iNOS activities of the EH group were 35.49±12.8 U/ml and 14.92±7.93 U/ml, and markedly lower than that 41.47±13.2 U/ml and 10.11±6.21U/ml in the healthy group (P<0.05). But the activities of eNOS in the EH group and healthy group were not significantly different (P>0.05).ConclusionThe variations of ecNOS4 gene locus may be responsible for the decrement of plasma NOx, both plasma NOx level and activity of NOS decreases in EH patients, so it may be a genetic susceptibility marker for EH of the Han nationality in China.

The use of animal medicine has a long history in China, it has the characteristics of high curative effect,strong activity, wide application and great potential. However,the circulation of animal medicine in current market mixed counterfeit variety and complex. Molecular identification technology of DNA barcoding is an emerging molecular biotechnology in recent years, it is a powerful supplement to traditional identification methods. This method can well identify animal species at the molecular level and has high accuracy, it can identify animal medicines quickly and monitor the medicine market effectively. This article summarizes the research process of molecular identification of DNA barcoding, the application of DNA barcoding in medicinal animals identification in recent years, and the limitations of DNA barcoding technology.
Animals ; China ; DNA ; DNA Barcoding, Taxonomic ; Research

Objective:To prepare standard decoction of Saposhnikoviae Radix pieces,and conduct a study on the quality standards, in order to provide the scientific reference for the study of clinical application and other decoction standard decoctions. Method:By the traditional water decocting method,11 batches of standard decoction of Saposhnikoviae Radix pieces were prepared to determine pH,prim-O-glucosylcimifugin and 4'-O-β-D-glucosyl-5-O-methylvisamminol content,calculate the paste rate and transfer rate,and investigate the feasibility of the measurement method. An analytical method for HPLC fingerprint was established. Hypersil ODS2 column (4.6 mm×250 mm,5 μm) was adopted and eluted with methanol-water as the mobile phase in a gradient mode. The detection wavelength was 254 nm,the flow rate was 1 mL·min^-1,and the column temperature was 25℃. The fingerprint analysis method was used to evaluate the similarity of the 11 batches of standard decoction of Saposhnikoviae Radix pieces. Result:Through the determination of 11 batches of standard decoctions of Saposhnikoviae Radix pieces,the average pH value was 5.5,the paste-out rates were between 34.3% and 46.3%,and the average paste-out rate was 41.4%,the standard deviation was 3.7%, the transfer rate of prim-O-glucosylcimifugin was 66.8%-93.5%,the average metastasis rate was 79.4%,the standard deviation was 12.1%, the transfer rate of 4'-O-β-D-glucosyl-5-O-methylvisamminol was 70.4%-98.2%,the average transfer rate was 83.40%,the standard deviation of 10.8%. The results of various methodological studies were consistent with the requirements of RSD. The fingerprinting analysis was performed using similarity evaluation software for traditional Chinese medicine fingerprints,and 9 principal common peaks were identified. The similarity was higher than 0.9. Conclusion:The quality evaluation method established in this study has a good stability,precision and repeatability. The fingerprints have a high similarity,which is suitable for the quality evaluation of standard decoctions of Saposhnikoviae Radix pieces, and can provide the scientific basis for the quality evaluation of other related preparations.

Depression is a mental or affective disorder， whose occurrence is closely related to the physical， psychological， and social environmental factors. Clinically， patients with depression often present with significant and long-lasting blue mood， impaired thinking and cognitive functions， reduced mobility， and even self-harm and suicide attempts. The pathogenesis of depression is complex and diverse， and there mainly exist the neurotransmitter hypothesis， immunodeficiency hypothesis， hypothalamus-pituitary-adrenal （HPA） axis activation hypothesis， brain-derived neurotrophic factor （BDNF） hypothesis， and intestinal flora hypothesis. In western medicine， it is mostly treated with antidepressant drugs， but the resulting side effects and repeated attack cannot be ignored. Traditional Chinese medicine（TCM） has obvious advantages in dealing with depression. The advocates of treating "Qi"， "phlegm"， "stasis"， and "five zang organs" have all been proved effective. Depression is characterized by a long course of disease and repeated attack， which coincides with the concept of "deficiency" in TCM. Tonifying deficiency is a method for treating depression based on TCM syndrome differentiation. Under the guidance of this principle， the formulated prescriptions will produce the desired effects. This article reviewed the related clinical and experimental studies on depression treatment via deficiency tonification in recent years， and summarized the corresponding prescriptions for benefiting Qi， replenishing blood， nourishing Yin， and tonifying yang， respectively， so as to enrich the TCM theory of depression and provide new ideas for its clinical treatment.

BACKGROUND: Blood platelet is active commonly appeared in both acute and recovery stages in ischemic cerebral apoplexy. Cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP) are closely correlated with the blood platelet function.OBJECTIVE: To observe the therapeutic effects of compound danshen drop on clinical prevention and treatment of transient cerebral ischemic attack (TIA) and the influence on serum cAMP and cGMP.DESIGN: Randomized control experiment.SETTING: Department of Neurology of Binzhou Medical College affiliatedHospitalPARTICIPANTS: Totally 180 TIA patients participated in the experiment in volunteer in the clinic of Neurological Department of Affiliated Hospital of Binzhou Medical College from September 2000 to April 2001. Based on randomized number scale, 3 groups were divided according to dosages of compound danshen drop. In 10 drops/d group, there were 60 cases, of which 35 cases were male and 25 cases female, aged varied from 50 to 70 years, averagely (54.3±7.2) years old. In 20 drops/d group, there were 60 cases, of which, 32 cases were male and 28 cases female, aged varied from 49 to 62 years, averagely (55.7±5.1) years old. In 30 drops/d group, there were 60 cases, of which, 33 cases were male and 27 cases female, aged varied from 52 to 69 years, averagely (54.9±5.5) years old.METHODS: In 10 drops/d group, the drug was given once per day, 10drops each time. In 20 drops/d group, the drug was given twice per day,10 drops each time. In 30 drops/d group, the drug was given three times per day, 10 drops each time. Four weeks after medication, 3 mL blood was collected from each patient in the morning. With radioimmunoassay, the contents of cAMP and cGMP in serum were determined. Each case was tions, gem bleeding, epistaxis, subcutaneous bleeding, mouth numbness,headache, dizziness, etc.tack: In 10 drops/d group, there were 1 case in internal carotid arterial system, 2 cases in vertebral arterial system, 1 case of cerebral infarction and 2 cases of myocardial infarction. In 20 drops/d group, it was 2, 2 and 1 cases successively and in 30 drops/d group, it was 1, 2, 1 cases succesgroup: There were 1, 2 and 4 cases of harmful accident and side effects in three groups successively, of which, peripheral numbness of mouth 1 case in 10 drops/d group, 1 case of either peripheral numbness of mouth or headache in 20 drops/d group, gastrointestinal reaction 1 case, peripheral numbness of mouth 2 cases and dizziness 1 case in 30 drops/d group.There was no gem bleeding, epistaxis and subcutaneous bleeding, and no patient was withdrawn the treatment due to above-mentioned side effects.With comparison among groups, significant difference was not indicated The contents of cAMP were (21.22±3.94), (22.5±3.67), (23.1±7.7) ng/Land cGMP were (3.67±1.18), (4.74±2.12), (4.6±0.7) ng/L successively in three groups (P ＞ 0.05).CONCLUSION: During second-grade prevention of TIA, compound danshen drop presents definite therapeutic effects, which is basically same in comparison of dosages. Simultaneously, no remarkable side effects appear.