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Objective:Due to the limitation of traditional identification methods of Chinese medicinal materials， the study established a rapid method to identify Persicae Semen mixed with Armeniacae Semen Amarum by allele-specific polymerase chain reaction （PCR）. Method:By comparing the ribosomal DNA internal transcribed spacer （ITS） gene sequences of Persicae Semen and Armeniacae Semen Amarum， single nucleotide polymorphism （SNP） sites were searched and specific primers were designed. Different Persicae Semen and Armeniacae Semen Amarum samples were amplified by PCR， the effects of annealing temperature， primer concentration and cycle number on the PCR reaction system were optimized， and the specificity and detection limit of this method were investigated. In addition， the established PCR method was used to detect the samples of Persicae Semen mixed with different proportion of Armeniacae Semen Amarum from different sources and producing areas. Result:A specific PCR method for identifying Persicae Semen mixed with Armeniacae Semen Amarum was established. When the annealing temperature was 63 ℃ and the number of primer cycles was 30， only Armeniacae Semen Amarum could be amplified with 432 bp specific band， while Persicae Semen samples did not have this band. The minimum detection limit of this method for Armeniacae Semen Amarum was 0.2 ng， and the detection limit for Armeniacae Semen Amarum adulterated in Persicae Semen was 1%. Conclusion:The established allele-specific PCR method can accurately detect whether there is Armeniacae Semen Amarum in Persicae Semen， which can provide experimental basis for the quality control of Persicae Semen and guarantee the safety of its clinical use.
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Textual research on Chinese herbs is the preliminary work for the preparation of famous classical prescriptions. Through literature review，it was found that the researches of Persicae Semen focused on chemical compositions，pharmacological mechanism and medical record analysis in the recent years，and few researches based on the ancient literature were about the origin，concocting methods，flavor，meridian tropism，effects and indications. Textual research shows that the most commonly used names are Taoren and Taoheren，with a wide range of producing areas. The plant origin of Persicae Semen has not changed much since ancient times. Before the Qing dynasty，the plant origin of Persicae Semen was from the seeds of a kind of fruit named Shantao or Maotao，and in modern times，the seeds of Amygdalus persica or A. davidiana have become the major source. While different books have different views on Latin names for the origin of the Persicae Semen. We suggest that the Latin names of A. persica and A. davidiana should be more reasonable for Tao and Shantao respectively .In the concocting methods of Persicae Semen for activating vital energy and blood circulation，raw Persicae Semen should be used with peel and tip，while for moisturizing dryness，it should be fried into yellow without peel. Therefore，in the concocting methods of Persicae Semen for Taohe Chengqitang and Taohong Siwutang，the raw materials should be fried into yellow without peel or tip，while for Shentong Zhuyutang，raw Persicae Semen materials should be used with peel and tip. The indications of Persicae Semen include amenorrhea，lump，parasite，obstruction of chest，cough and asthma，constipation，etc.，and the people with blood deficiency，blood dryness and the pregnant women should use it with caution or should not use it. The modern clinical application of Persicae Semen is only a partial inheritance of ancient literature，which means that the Persicae Semen still has many effects to be verified and studied，and it is worthwhile for further exploration in order to expand its clinical application. The records of ancient literature on flavor，meridian tropism，and quality evaluation of Persicae Semen were consistent with those in Chinese Pharmacopoeia. Because of the similar appearance，it is especially difficult to distinguish Persicae Semen and Armeniacae Semenis Amarum after crushing ，requiring much time and money for identification. It is recommended that medical institutions should purchase Persicae Semen without crushing as far as possible，then decide the best concocting methods according to the clinical requirement.
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In order to establish a rapid and non-destructive evaluation method for the identification of Armeniacae Semen Amarum and Persicae Semen from different origins, the spectral information of Armeniacae Semen Amarum and Persicae Semen in the range of 898-1 751 nm was collected based on hyperspectral imaging technology. Armeniacae Semen Amarum and Persicae Semen from different origins were collected as research objects, and a total of 720 Armeniacae Semen Amarum samples and 600 Persicae Semen samples were used for authenticity discrimination. The region of interest(ROI) and the average reflection spectrum in the ROI were obtained, followed by comparing five pre-processing methods. Then, partial least squares discriminant analysis(PLS-DA), support vector machine(SVM), and random forest(RF) method were established for classification models, which were evaluated by the confusion matrix of prediction results and receiver operating characteristic curve(ROC). The results showed that in the three sample sets, the se-cond derivative pre-processing method and PLS-DA were the best model combinations. The classification accuracy of the test set under the 5-fold cross-va-lidation was 93.27%, 96.19%, and 100.0%, respectively. It was consistent with the confusion matrix of the predicted results. The area under the ROC curve obtained the highest values of 0.992 3, 0.999 6, and 1.000, respectively. The study revealed that the near-infrared hyperspectral imaging technology could accurately identify the medicinal materials of Armeniacae Semen Amarum and Persicae Semen from different origins and distinguish the authentication of these two varieties.
Subject(s)
Drugs, Chinese Herbal , Hyperspectral Imaging , Least-Squares Analysis , Semen , Support Vector Machine , TechnologyABSTRACT
Objective: To explore the prescription medication rule of the first batch of Chinese medicine masters in treating acute stage of stroke by using the data mining method. Methods: The experience books and published journal articles of Chinese medicine masters and the shared medical records of national famous TCM studios in the cloud platform V1.5 of ancient and modern medical records were searched; The first batch of national physician master data for the treatment of acute phase of stroke was selected; A standardized basis database, standardization of medicines were established after using statistical analysis of system integration, association rules analysis, methods of complex networks; The formula frequency, medicinal, and core prescription drugs were analyzed. Results: A total of 142 medical cases were eventually included. The frequency results of traditional Chinese medicine showed that 14 kinds of high-frequency traditional Chinese medicine were obtained, including Pheretima, Chuanxiong Rhizoma, Persicae Semen, etc., and most of the drugs were warm, smooth, bitter, sweet, and return to liver and lung meristem. The results showed that the frequency of chordal pulse and tongue red was the most. A total of 24 association rules and 18 TCM association rules were obtained by association rule analysis. Conclusion: In the treatment of acute apoplexy, the Chinese medicine master usually adopted the methods of clearing heat and relieving wind, promoting blood circulation and removing blood stasis, eliminating phlegm and resuscitative medicinal, moving qi, and dredging collaterals, and stopping convulsions.
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Objective: A Logistic model for quality evaluation of Persicae Semen slices was constructed and its feasibility was verified in this study based on the thoughts on quality control of Chinese materia medica “Components reflect activity and activity points to efficacy”. Methods: Content of amygdalin, thrombin time (TT), ABTS clearance rate, DPPH inhibition rate, and hydroxyl radical scavenging ability in vitro were analyzed by a binary Logistic algorithm. Besides, a grade prediction model for Persicae Semen was established and verified. Results: A total of 18 batches of Persicae Semen were divided into four grades: excellent (represented by Neimeng Persicae Semen), good (represented by Gansu Persicae Semen), medium (represented by Liang Persicae Semen), and poor (represented by Shaanxi Persicae Semen) according to the probability formula of the Logisitc model. All batches of Persicae Semen slices were classified accurately, manifested by the high value of fitting probability (P > 98%). Conclusion: The classification standard based on the Logistic algorithm involving quality control component-in vitro titer is applicable to Persicae Semen slices on the market. Classification results are accurate and reliable.
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Persicae semen has been used for years as a traditional Chinese medicine to treat diseases. Because of their similar morphologies, Persicae semen was commonly inadvertently mixed with Armeniacae semen amarum (a toxic herbal seed). Development of a reliable method for discriminating Persicae semen from its adulterant is necessary to reduce confusion for the drug safety in clinical practices. This study evaluates the efficiency of high-resolution melting (HRM) combined with internal transcribed spacer 2 (ITS2) to analyze Persicae semen. Our findings show that HRM allows not only the identification of adulteration but also the quantification of the most common admixture. HRM sensitivity in adulterant detection was assessed through the analysis of mixing samples with different proportions of Prunus persica and Prunus armeniaca control. The results are presented as a linear regression with r of 0.96 and imply the capability of the method to detect adulteration. In particular, HRM detected seeds of Prunus persica in Prunus armeniaca at concentrations as low as 1%, and commercial products labeled as ‘Persicae semen’ were purchased from markets and could rapid authenticated by HRM analyses. This study is significant in the verification of the authenticity in the quality control of herbal medicine. In the near future, it is promising to be the main trend for identifying traditional Chinese medicinal materials.
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Objective To evaluate Persicae Semen-Rhei Radix et Rhizoma (PS-RRR) of different compatibility of acute blood stasis rats blood rheology and blood coagulation function, and to reveale the effect of PS-RRR for promoting blood circulation to remove blood stasis effect scope, nature, and degree of interaction. Methods With ice water bath and injected adrenaline hydrochloride copy of acute blood stasis rats model, different ratio (0:1, 1:5, 2:5, 2:3, 1:1, 3:2, 5:2, 5:1, and 1:0) of different concentration of PS-RRR was given for later. Through the determination of whole blood viscosity (WBV), blood sedimentation (ESR), erythrocyte aggregation index (EAI), prothrombin time (PT), partial thrombin (APTT), fibrinogen (FIB) content of coagulation, thrombin time (TT), and the blood rheology of blood stasis rats was observed, the influence of the blood coagulation indexes. Then response surface analysis and multi-index comprehensive index method of PS-RRR different compatibility of promoting blood circulation was used to remove blood stasis effect comprehensive comparative analysis. Results The ratio of PS-RRR between 2:3 to 3:2 showed obvious synergy (strength of synergy: -0.8); In PS dose from 5.5-10 g and RRR dose from 2.1-5.8 g area showed the antagonism function (antagonism effect strength maximum: 0.6); While other percentage did not show obvious synergy or antagonism. Conclusion The results of reveal that scope, nature, and degree of PS-RRR for promoting blood circulation to remove blood stasis interaction effect, the and its prescriptions of traditional Chinese medicine (TCM) and clinical use of PS-RRR 1:1 is consistent with the highest frequency of conclusion. PS-RRR provides scientific basis for clinical applications.
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The aim of this review is to summarize the advances in studies on Persicae Semen, a Chinese herbal medicine, in the fields of nature and flavor, preparation, quality control, chemical composition, pharmacology, and toxicology. Persicae Semen and its extracts present various pharmacological activities, such as cardiovascular protection, neuroprotection, immunoregulation, antitumor effects and hepatic and renal protection. Though amygdalin, one of the main components in Persicae Semen, had been well studied, the activities and mechanisms of other potential active ingredients should also be more concerned.
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Objective:To study the effect of different dose of persicae semen extract extract(PSE) to barrier function of the intestinal mucous membrane and immunologic function in acute pancreatitis rats.Methods:A total of 48 rats were divided into model control group,low dose,medial dose and high dose PSE groups,and there were 12 rats in each group.Another 12 rats were Sham-operation group.After anesthesia recovery,rats in low dose,medial dose and high dose PSE groups respectively received PSE 0.12 g/kg,0.248 g/kg and 0.36 g/kg,and rats in Sham-operation group and model control group receive isovolumetric distilled water,once per 6 h,4 times in 24 hours.All rats were anesthetized by 10%chloral hydrate after in 24th hour after dosing.Thorax and enterocoelia were opened; 5 ml of blood were respectively drawed to EDTA-anticoagulation tube and un-anticoagulation tube from aorta abdominalis.CD4+, CD8+and Treg cells were determined by direct fluorescent-labelded flow cytometry.IgA, IgG and IgM were determined by immunoturbidimetry.Serum amylase was determined by EPS-G7 substrate,D-lactic acid was determined by enzymology, and serum diamine oxidase was determined by active ration of colorimetry method.Pathological examination of small intestine mucous membrane tissue was taken after HE staining.sIgA in small intestine was determined by radioimmunoassay.mRNA of TLR4 and NF-κBp65 in small intestine tissue was determined by RT-PCR.Results:(1) Serum amylase,D-lactic acid and diamine oxidase in medial dose and high dose PSE groups were significantly decreased ( P<0.01 ) , and sIgA in small intestine was significantly increased ( P<0.01).These indicators were significantly different in medial dose and high dose PSE groups(P<0.01).(2) CD4+and CD4+/CD8+in medial dose and high dose PSE groups were significantly increased(P<0.01),and CD8+,Treg cells were significantly decreased(P<0.01) compared with those in low dose PSE group.These indicators were significantly different in medial dose and high dose PSE groups(P<0.01).(3) IgA,IgG and IgM in medial dose and high dose PSE groups were significantly decreased(P<0.01) compared with those in low dose PSE group.These indicators were significantly different in medial dose and high dose PSE groups(P<0.01).(4) Small intestine mucous membrane tissue in Sham-operation group was not damaged significantly,but that in model control group was damaged significantly.Small intestine mucous membrane tissue in low dose PSE group was similar to that in model control group,and damage in medial dose and high dose PSE groups was decreased significantly.( 5 ) mRNA of TLR4 and NF-κBp65 in small intestine tissue in medial dose and high dose PSE groups were significantly increased ( P<0.01 ) compared with those in low dose PSE group.These indicators were significantly different in medial dose and high dose PSE groups ( P<0.01 ).Conclusion: PSE has protective effect to barrier function of the intestinal mucous membrane,and significantly improve the immunologic function.
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Objective: To study the different pharmacodynamic effects of Persicae Semen extract (PSE) on the blood circulation disorder of rats induced by cold congeal and blood stasis syndrome (cold syndrome, CS), and heat congeal and blood stasis syndrome (heat syndrome, HS). Methods: Eighty rats were randomly divided into eight groups including normal control group of CS, model group of CS, PSE (8 g/kg) group of CS, Chanxiong Rhizoma (8 g/kg) group of CS, normal control group of HS, model group of HS, PSE (8 g/kg) group of HS, and Salviae Miltiorrhizae Radix (14 g/kg) group of HS. The two rat models were established by freezing and injection of carrageenan solution. The rats in each group were ig administered once daily for 7 d. The blood flow velocity (Fve) and blood viscosity (Vis) were detected; The organs (such as heart, lung, liver, kidney, and spleen) were taken and observed by histopathological analysis; The diameters of tiny blood vessels were measured by pathology image analysis software, and the thrombosis and tissue injury were observed. Results: PSE increased Fve but decreased Vis in CS and HS rats (P < 0.05), made the small artery in CS rats systolic, while that in HS rats diastolic (P < 0.05). The organ injury severity score indicated PSE could attenuate the injury of many organs (such as heart, lung, liver, and kidney) in CS rats, but only attenuated the injury of kidney in HS rats (P < 0.05). Conclusion: PSE could improve the blood circulation disorder induced by CS and HS. The different effects of PSE on the two kinds of syndromes include the regulation of the diastolic and systolic states of rat small artery and the different protection of rat organs.
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Objective: To develop a precolumn saponification HPLC method for determining linolic acid and oleic acid in Persicae Semen from different habitats and to provide the basis for controlling the herb quality quickly and accurately. Methods: The linolic acid and oleic acid in Persicae Semen were saponified with 0.5 mol/mL KOH/EtOH as saponifier. HPLC method was used to determine the contents of linolic acid and oleic acid. The column was Waters-Symmetry-RP-C18 (250 mm × 4.6 mm, 5 μm), the mobile phase consisted of acetonitrile-0.1% aqueous phosphoric acid (92:8); The flow rate was 1.0 mg/mL; The absorbance was monitored at 205 nm; The column temperature was 30°C. Results: The calibration curves of linolic acid and oleic acid were in a good linearity over the ranges of 8.193 2-163.864 μg/mL (r = 0.999 7) and 26.4-528.0 μg/mL (r = 0.9995), and the average recoveries of linolic acid and oleic acid were 97.3% and 98.0% with RSD values of 2.7% and 2.3%, respectively (n = 6). Conclusion: This method is simple, sensitive, and accurate, which is suitable for the simultaneous determination of linolic acid and oleic acid in Persicae Semen.
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Objective: To identify Persicae Semen and its sibling species, and to secure the quality and clinical safety of this medicinal material. Methods: The internal transcribed spacer 2 (ITS2) of nuclear ribosomal DNA of Persicae Semen and its sibling species were amplified and bidirectionally sequenced by DNA barcoding. Sequence assembly and consensus sequence generation were performed by the CodonCode Aligner 4.1. The genetic distances were computed by MEGA 5.0 in accordance with the Kimura 2-parameter (K2P) model, and the phylogenetic tree constructed by the neighbor-joining (NJ) method. Results: The length of ITS2 sequences of the two origin plants of Persicae Semen was between 212 bp to 213 bp. Their intraspecific genetic distance was much lower than the interspecific genetic distance with their sibling species. The ITS2 sequence possessed the character of good stability and low intra-specific sequence variation. In the NJ tree, both Prunus persica and P. davidiana were clustered into one large branch, and clearly separated with their sibling species. Conclusion: ITS2 can be used to effectively distinguish Persicae Semen from its sibling species, which can provide a reference for the iden-tification of other Chinese medicine and its sibling species.