1.Characteristic Analysis of Imprinting Template of Flavonoid Clusters in Four Chinese Medicines to Lung Meridian and Establishment of Experimental Approach of Meridian Tropism in Vitro
Qijun HE ; Dandan SHENG ; Yuxia CHEN ; Shaoqin OUYANG ; Kaiwen DENG ; Fuyuan HE ; Xinyu CHEN
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(9):141-147
ObjectiveTo study the characteristics of imprinting template of flavonoid clusters in four Chinese medicines attributed to the lung meridian, and to establish an in vitro experimental approach for the study of the attribution of Chinese medicines to the lung meridian. MethodBased on 13 Chinese medicines, including Xanthii Fructus, Houttuyniae Herba, Fagopyri Dibotryis Rhizoma, Belamcandae Rhizoma and so on, which only belong to the lung meridian in Chinese Materia Medica(the 13th Five-Year planning textbook of general higher education), we identified four representative Chinese medicines, namely Houttuyniae Herba, Fagopyri Dibotryis Rhizoma, Belamcandae Rhizoma and Mori Cortex, and set up their fingerprints by high performance liquid chromatography(HPLC) and calculated the molecular connectivity indices of various components in the four Chinese medicines, the similarity to their mean value was calculated by included angle cosine method, so as to establish the quantitative relationship of construction versus imprinting ability, and to determine the order of each component in the lung meridian. A total of 7 reference substances, including chlorogenic acid, rutin, quercetin, isoquercitrin, hyperoside, epicatechin, and iridin, were selected to validate the overall conformational relationships of flavonoids of the model, as well as its predictive ability. ResultHouttuyniae Herba, Fagopyri Dibotryis Rhizoma, Belamcandae Rhizoma and Mori Cortex contained a total of 437 chemical components with an average molecular connectivity index similarity of 0.995 6. The four Chinese medicines contained a total of 204 flavonoids with an average molecular connectivity index similarity of 0.978 0, which was second only to the alkaloids with 0.985 1. The retention time(tR) of the 7 reference substances showed a good conformational relationship with the similarity of the molecular connectivity index(tR=831.4×S-790.3, r=0.861 4, P<0.01), which was applicable to the in vitro attribution study of the position, similarity, and relative similarity with tR of the cluster of 98.04% of flavonoids. Accordingly, the 1st position was kuwanon D, with a similarity of molecular connectivity index of 0.987 7 and a tR of 30.88 min, the 200th position was chlorogenic acid, with a similarity of molecular connectivity index of 0.958 2 and a tR of 6.36 min. The total first-order moment of the four Chinese medicines calculated by total statistical moment method of fingerprint was 24.26 min, ranked 21, which could characterize 99.19% of the whole, and the total first-order moment of the total peak area of the 7 reference substances in the four Chinese medicines was 20.00 min, with a rank of 46, which could characterize 98.68% of the whole. ConclusionFlavonoid clusters are suitable probes for the characterization of imprinting template for the study of the lung meridian, which can be established a quantitative imprinting method for meridian tropism of Chinese medicines in vitro.
2.Establishment of rapid detection method for zika virus based on direct amplification RT-PCR technique
Lang LI ; Libing GU ; Li ZHU ; Jianan HE ; Ying YE ; Ran ZHANG ; Huawen LI ; Fuyuan LI ; Dayong GU
International Journal of Laboratory Medicine 2024;45(3):358-364
Objective To establish a rapid detection method for zika virus based on direct amplification re-al-time fluorescent quantitative reverse transcription polymerase chain reaction(RT-PCR)technique.Methods A direct amplification RT-PCR technique for the rapid detection of zika virus in 5 samples(whole blood,serum,saliva,throat swab and urine)was established by using a special function DNA polymerase and a preferred PCR enhancer.Results The detection limits of the 5 samples were 103 PFU/mL in serum,102 PFU/mL in urine,throat swab,and saliva,and 104 PFU/mL in whole blood.The coefficient of goodness-fit of stand-ard curves was above 0.98,and the amplification efficiency was 90%-110%.Zika virus nucleic acid was suc-cessfully amplified,but non-zika virus nucleic acid was not amplified.Based on the repeatable detection of sam-ples from urine,whole blood,and saliva,the variation coefficient of 6 repeated Ct values at 106 PFU/mL and 102 PFU/mL concentrations were all<5%.The zika virus detection method established by the direct amplifi-cation RT-PCR technique was consistent with the detection results of conventional RT-PCR technique.Only two serum samples were detected in eight zika virus samples,and the remaining 62 non-zika virus samples and 12 negative samples were not amplified.Conclusion A rapid detection method for zika virus based on direct ampli-fication RT-PCR technique is successfully established.The method is simple,rapid,sensitive and specific.
3.Research on Material Basis of Homologous and Heterogeneous Effect of Aurantii Fructus Immaturus and Aurantii Fructus Based on Total Statistical Moment Analysis and Molecular Connectivity Index
Xiao YUAN ; Yuefeng WU ; Shixiong LI ; Youzhi LIU ; Zhiqin YU ; Fuyuan HE ; Jin ZHOU
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(3):161-168
ObjectiveTo investigate the material basis of homologous and heterogeneous effect of Aurantii Fructus Immaturus(AFI) and Aurantii Fructus(AF) based on the total statistical moment analysis and molecular connectivity index(MCI). MethodRelevant literature at home and abroad and Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP) were consulted to establish the chemical composition database of AFI and AF, and set up their fingerprints by ultra-high performance liquid chromatography(UPLC), and the total statistical moments and similarity parameters of the fingerprint were calculated. According to MCI, all components of AFI and AF were divided into different component groups, the average values of 0-8th order(0χ-8χ) MCI of the common component groups of AFI and AF were calculated. ResultThe values of total zero-order moment(AUCT) of AFI and AF were (10.57±2.45)×106, (5.09±0.89)×106 μV·s, the values of total first-order moment(MCRTT) were (11.57±1.58), (12.10±1.29) min, the values of total second-order moments(VCRTT) were(24.49±2.30), (26.49±2.54) min2, respectively. It showed that qualitative and quantitative parameters of AFI and AF were significantly different. The components with high similarity such as neohesperidin, hesperidin and narirutin were screened as the common potential pharmacodynamic components of AFI and AF. The non-common components of AFI, such as alysifolinone and imperatorin, and the non-common components of AF, such as neoeriocitrin and isosakuranin, with high similarity were screened out as potential heterogeneous components of AFI and AF. The composition groups of AFI and AF were classified into six categories, and the similarities between the composition groups of AFI and AF and the total constituents were 0.872-0.979 and 0.918-0.997, the average values of 0χ-8χ MCI of alkaloids in AFI and AF were 3.65 and 3.14, the average values of 0χ-8χ MCI of flavonoids were 8.47 and 8.47, the average values of 0χ-8χ MCI of volatile oils were 2.71 and 3.48, respectively. It showed that there were some differences in MCI of chemical constituents(groups) between AFI and AF. ConclusionThe chemical constituents(groups) of AFI and AF not only differ in content and species, but also in structural characteristics and structure-activity relationship, which can provide a basis for further explaining the scientific connotation of homologous and heterogeneous effect of AFI and AF.
4.A novel method for integrating chromatographic fingerprint analytical units of Chinese materia medica: the matching frequency statistical moment method
LI Haiying ; PAN Xue ; WANG Mincun ; LI Wenjiao ; HE Peng ; HUANG Sheng ; HE Fuyuan
Digital Chinese Medicine 2024;7(3):294-308
Methods:
This study established the MFSM method. To demonstrate its effectiveness, we applied this novel approach to analyze Danxi Granules (丹膝颗粒, DXG) and its constituent herbal materials. To begin with, the ultra-performance liquid chromatography (UPLC) was applied to obtain the chromatographic fingerprints of DXG and its constituent herbal materials. Next, the MFSM was leveraged to compress and integrate them into a new fingerprint with fewer analytical units. Then, we characterized the properties and variability of both the original and integrated fingerprints by calculating total quantum statistical moment (TQSM) parameters, information entropy and information amount, along with their relative standard deviation (RSD). Finally, we compared the TQSM parameters, information entropy and information amount, and their RSD between the traditional and novel fingerprints to validate the new analytical method.
Results:
The chromatographic peaks of DXG and its 12 raw herbal materials were divided and integrated into peak families by the MFSM method. Before integration, the ranges of the peak number, three TQSM parameters, information entropy and information amount for each peak or peak family of UPLC fingerprints of DXG and its 12 raw herbal materials were 95.07 − 209.73,
5.Network Correlation Analysis Between Components of Shuanghuanglian Injection and Allergy-like Targets
Weilong ZHANG ; Hong HE ; Ru QIAO ; Peng HE ; Wenjiao LI ; Liangqi ZHANG ; Xiaoxuan LIU ; Siqi HUANG ; Xue PAN ; Fuyuan HE
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(19):190-197
ObjectiveBased on the network pharmacology system and quantitative spectroscopy of traditional Chinese medicine(TCM) compounds, a topological network analysis method with equilibrium constant as the core was established to further explore the interaction between allergenic components and their network targets in Shuanghuanglian injection(SHLI), in order to provide new ideas and experimental basis for identifying and screening potential allergens of SHLI. MethodAfter one week of adaptive feeding, 72 SPF-grade SD male rats were randomly divided into blank group, SHLI standard group, Lonicerae Japonicae Flos(LJF) group, Scutellariae Radix(SR) group, Forsythiae Fructus(FF) group, and 7 groups of SHLI matching groups(groups 1-7), with 6 rats in each group. Rats in each group were administered the drug intravenously and blood samples were taken after steady state, high performance liquid chromatography(HPLC) characterization profiles of the testing drugs and plasma components in each group were established, and the peak area changes of the drugs and plasma components in each group were calculated after the component groups were classified. Enzyme-linked immunosorbent assay(ELISA) was used to determine the changes of immunoglobulin E(IgE), histamine(HIS), tryptase(TPS), total complement(CH50) and terminal complement complex(C5b-9) in animal blood samples. MATLAB R2020b v9.9.0 software was used to calculate the network balance constants of the component groups with the targets, and the eigenvalues of the matrices composed of network equilibrium constants were calculated and ranked according to their values. ResultELISA results showed that, compared with the blank group, groups 1-3 could significantly increase the IgE level, groups 1-2, groups 4-6 and SHLI standard group could significantly increase the HIS level, group 4 could significantly increase the CH50 level, groups 1, 3-4, LJF group and FF group could significantly increase the TPS level, SR group could significantly increase the C5b-9 level, and the differences were all statistically significant(P<0.05). According to the retention time of chromatographic peaks, it was classified into 6 component groups from C1 to C6 by HPLC. The order of the network balance constants of each component group was C6>C4>C1>C5>C3>C2, indicating that C6 had the greatest effect on the allergic reaction, and was most likely to be the allergen. The sequence of eigenvalues was C2>C5b-9>C3>C1>CH50>C6>C5>IgE>TPS>C4>HIS, indicating that component group C2 had the greatest contribution to the whole network. ConclusionBased on the correlation analysis of SHLI component group and allergy-like target network, this study clarified that component group C6 may be a potential allergen in SHLI, and the component group C2 may be a key node in the mechanism of drug action, which can provide new strategies and methods for the screening of allergens in TCM injections.
6.Theory and Analysis of Pharmacokinetic and Chromatokinetics Dose-time Characterization Methods in Traditional Chinese Medicine
Ru QIAO ; Peng HE ; Qijun HE ; Haiying LI ; Meifeng XIAO ; Kaiwen DENG ; Fuyuan HE
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(22):178-186
ObjectiveTo establish a theoretical system of pharmacokinetic and spectrokinetic dose-time characterization of traditional Chinese medicine(TCM). By analyzing the pharmacokinetic and spectrokinetic behaviors of Lonicerae Flos, Houttuyniae Herba injection, Lonicerae Japonicae Flosand Buyang Huanwutang, this paper compared the similarities and differences of the three methods for characterizing the dose-time relationship, namely half-life, statistical moment and statistics, in order to find the most suitable method for characterizing the relationship. MethodTen mice were randomly selected from 100 Kunming mice as the blank group, and the remaining mice were coated with xylene in the auricle to establish the acute inflammation model of ear swelling. After successful modeling, the mice were gavaged with aqueous extract of Lonicerae Flos(30 g∙kg-1), and the blank group was gavaged with an equal volume of normal saline. The plasma of mice was collected at different time points to determine the content changes of components. At the same time, the pharmacokinetic results of Houttuyniae Herba injection, Lonicerae Japonicae Flos and Buyang Huanwutang were included, and the pharmacokinetic and spectrokinetic parameters were calculated. Then the difference in the time of calculating 95% total component content of metabolism by half-life method, statistical moment method and statistical method was compared. ResultOn the basis of the half-life method, the mathematical expressions of statistical moment method and statistical method suitable for the characterization of dose-time relationship of multi-component system of TCM were established. The results showed that the pharmacokinetic parameters of the individual components in Lonicerae Flos varied, with cryptochlorogenic acid and rutin showing a two-compartment model and the other components showing a one-compartment model. After calculation of spectrokinetic similarity, the metabolic patterns among the components contained in Houttuyniae Herba injection, Lonicerae Japonicae Flos, Lonicerae Flos and Buyang Huanwutang were different and varied greatly in vivo. The time to metabolize 95% of the total components of the four research subjects in vivo was calculated by the half-life method, statistical moment method and statistical method, and it was found that the difference between statistical moment method and half-life method was large, and the difference between statistical moment method and statistical method was small. ConclusionStatistical method not only reflects the characteristics of statistical moment method, characterizes the dispersion degree of each component, but also can be associated with fingerprint to form spectrokinetics, characterizing the dose-time relationship of 95% of drug components, which is a more desirable method to characterize the dose-time relationship of the component groups in TCM.
7.Study on the Characteristics of Supramolecular"Imprinting Template"of Curcuma kwangsiensis and Curcuma phaeocaulis
Weilong ZHANG ; Yuchai WANG ; Ru QIAO ; Peng HE ; Shanshan LIANG ; Sheng LIU ; Yingyang TAN ; Yu TANG ; Limin GONG ; Fuyuan HE ; Xue PAN
Chinese Journal of Information on Traditional Chinese Medicine 2024;31(6):123-129
Objective The characteristics of supramolecular"imprinting template"of volatile oil of Curcuma kwangsiensis and Curcuma phaeocaulis were analyzed and studied based on the supramolecular"qixi"theory of Chinese materia medica combined with chemometrics.Methods The volatile oil of C.kwangsiensis and C.phaeocaulis were extracted by steam distillation,and the fingerprint and composition information of each batch were obtained by GC-MS.Total statistical moment method was used to compare the imprinting characteristics of the"imprinting template"of C.kwangsiensis and C.phaeocaulis.The core index(CI)of each batch of essential oil of C.kwangsiensis and C.phaeocaulis was calculated,and the topological characteristics of types of"imprinting template"of C.kwangsiensis and C.phaeocaulis were compared by chemometrics.Results There was no significant difference in the extraction rate of volatile oil between C.kwangsiensis and C.phaeocaulis.The average values of total zero order moment(AUCT)were(1.907±0.177)×108,(1.979±0.413)×108 μV·s,respectively,showing that there was no significant difference in the total content of volatile oil between the two groups.The mean values of the total first order moment(MCRTT)were(30.969±0.962)and(33.198±0.409)min.The average value of total second order moment(VCRTT)was(56.176±11.368)and(43.891±4.113)min2,respectively,indicating that there were significant differences in the content ratio and species of volatile oils between the two groups.The similarity of total statistical moments of C.kwangsiensis and C.phaeocaulis was mostly lower than the defined value,indicating that the chemical composition and composition ratio of the volatile oil were different.Principal component analysis and orthogonal partial least squares discriminant analysis could obviously divide C.kwangsiensis and C.phaeocaulis into two categories.Through the analysis of P value and VIP value,the CI values of Xvp 4th order,Xvpc 5th order,Xvpc 6th order,Xvpc 7th order,Xvc 3rd order,Xvpc 4th order were the main difference values of C.kwangsiensis and C.phaeocaulis.Conclusion Through the characterization of"imprinting property"and"topological characteristics"of the supramolecular"imprinting template"and combining with chemometric analysis,it is possible to successfully distinguish C.kwangsiensis and C.phaeocaulis,and find the different CI values between two"imprinting templates".
8.Workplace violence and its impact on occupational burnout among healthcare workers in an infectious disease hospital
Yuting TANG ; Min ZHANG ; Dongmei LIU ; Yiming HUANG ; Chuning HE ; Fuyuan WANG ; Hongli TANG
China Occupational Medicine 2023;50(5):578-584
{L-End}Objective To explore the impact of workplace violence (WPV) on occupational burnout among healthcare workers. {L-End}Methods A total of 675 healthcare workers from an infectious disease hospital were selected as the study subjects using typical sampling method. The Workplace Violence in the Healthsector Country Case Studies Research Instruments: Survey Questionnaire and the Maslach Burnout Inventory 2-item were used to investigate the incidence of WPV and occupational burnout. {L-End}Results The incidence of WPV among the study subjects was 35.1%, with incidences of physical and psychological violence at 2.2% and 34.1%, respectively. The detection rates of emotional exhaustion, depersonalization, and occupational burnout were 25.9%, 12.6%, and 52.4%, respectively. The result of multifactorial logistic regression analysis showed that experiencing psychological violence, having a bachelor or master degree or higher, and having more concerns about WPV were influencing factors for emotional exhaustion (all P<0.05). Knowing the reporting process for violent incidents and having more concerns about WPV were influencing factors for depersonalization (all P<0.05). Being in a minority ethnic group, having a bachelor, a master degree or higher, experiencing psychological violence, and having more concerns about WPV were influencing factors for occupational burnout (all P<0.05). {L-End}Conclusion WPV increases the risk of occupational burnout among healthcare workers. Effective measures should be implemented to reduce the incidence of WPV, decrease the level of occupational burnout, and promote the overall well-being of healthcare workers.
9.Effects of carbonization processing on quality control, chemical compositions, and pharmacological mechanism of Ganjiang (Zingiberis Rhizoma)
Yiqun ZHOU ; Ping WU ; Yu TANG ; Wenlong LIU ; Jilian SHI ; Fuyuan HE
Digital Chinese Medicine 2023;6(3):341-356
Ganjiang (Zingiberis Rhizoma, ZR) and Jiangtan (Carbonized Zingiberis Rhizoma, CZR) have
long been used in traditional Chinese medicine (TCM) with a rich history in the treatment of
various ailments. While ZR and CZR obviously stem from the same botanical source, their attributes, chemical compositions, pharmacological behaviors, and clinical applications are different owing to variations in the extent of drying and processing they undergo. In this paper,
data pertaining to ZR and CZR were retrieved from databases including China National
Knowledge Infrastructure (CNKI), PubMed, Web of Science, and Google Scholar. These
sources were scrutinized to elucidate the distinctions between ZR and CZR arising from carbonization processing in terms of their ethnopharmacology, quality control, chemical compositions, biological activities, pharmacological mechanisms, and clinical uses. In this study,
a total of 56 chemical constituents were identified and isolated from ZR and CZR, which primarily encompassed volatile oils, gingerols, and diphenylheptane compounds. CZR's pharmacological effects include hemostatic, anti-oxidant, analgesic, antibacterial, anti-cancer,
and other biological activities. ZR has pungent and warm properties. It is a Yang-supplementing herbal medicine for ailments exacerbated by cold or damp climatic influences. CZR is a
product of ZR after undergoing high temperature, with diminished intensity of its pungent
and warm attributes. This change leads to a more gradual treatment efficacy, renowned
hemostatic effects and its ability to gently invigorate the spleen and effectively alleviate diarrhea. Currently, research on the pharmacological mechanism of CZR is mainly focused on the
effects of CZR on coagulation and fibrinolysis. Although the healing effect of CZR has long
been known, and some correlation has been found between the changing composition and
the changing color of the decoctions, people still lack relatively clear processing mechanisms
to reflect the characteristics and specific quality standards of the ingredients of CZR's hemostatic effect. This review provides a systematic summary on quality control, chemical composition, ethnopharmacology, and pharmacology of CZR, offering novel perspectives for advancing the exploration of additional carbonized herbal medicine and fostering their application in clinical settings
10.Chemical Constituents and Insulin Resistance Targets in Different Parts of Morus alba by UPLC-Q-TOF-MS Combined with Network Pharmacology
Yuefeng WU ; Jinling LIU ; Dingfang CHEN ; Zhifei ZHU ; Youzhi LIU ; Xiao YUAN ; Jin ZHOU ; Fuyuan HE
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(2):149-158
ObjectiveTo compare the similarities and differences of material basis for improving insulin resistance (IR) in different parts of Morus alba based on liquid-mass combination combined with network pharmacology and molecular docking technology. MethodUltra-high performance liquid chromatography tandem quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) was used to analyze the composition differences in different parts of M. alba. Sybyl-X2.1 was used to connect components with IR core targets, and the selection criterion was Total Score≥5. The "component-target-disease" network map was drawn. The total statistical moment standard similarity (TQSMSS) between the single target-component docking score data set and the total target-component docking score data set was calculated. The targets with higher TQSMSS were screened out, and the protein-protein interaction (PPI) network was constructed. The Gene Ontology (GO) functional analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis were performed using R language. ResultForty-one active components were obtained by UPLC-Q-TOF-MS. According to the total statistical moment (similarity) method, there were 20, 23, 30, and 27 targets with TQSMSS≥0.75 in Mori Ramulus, Mori Cortex, Mori Fructus, and Mori Folium, respectively. In the four M. alba medicinal sources, the functional order of the targets by GO enrichment analysis was Mori Fructus>Mori Folium>Mori Cortex>Mori Ramulus, which were involved in biological processes such as blood glucose homeostasis, glucose metabolism, and glucose transmembrane transport. The order of the four M. alba medicinal sources by KEGG pathway enrichment analysis was Mori Fructus>Mori Ramulus>Mori Folium>Mori Cortex, which were involved in the adenosine monophosphate-activated protein kinase (AMPK) energy metabolism signaling pathway, the insulin regulation-related signaling pathway, the anti-inflammatory and anti-oxidative stress signaling pathway, and so on. ConclusionThis research demonstrates that there are differences in the material basis for improving IR by different parts of M. alba, which provides references for the development of different parts of M. alba.

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