Modern technology-based exploration of mechanism of traditional Chinese medicine in prevention and treatment of gastric cancer

This review comments on the article“To explore the mechanism of Yigong San anti-gastric cancer and immune regulation”.We are interested that the article applied network pharmacology and bioinformatics techniques to elucidate the mechanism of action of Yigong Sang,a traditional Chinese medicine(TCM),in the treatment of gastric cancer(GC).The mechanism of action of Yigong Sang in the treatment of GC has not yet been elucidated because it is composed of multiple Chinese medicines with multiple components and multiple targets.The emergence of network pharmacology and bioinformatics analysis helps explain the mechanism of action of TCM in preventing and treating GC,and provides a possibility for TCM to transform from empirical to evidence-based medicine.This is of great significance for the application of TCM in oncology,new drug development,formula optimization,and the improvement of clinical efficacy.

Visual Analysis of the Application and Development Trend of Cinnamon in the Field of Traditional Chinese Medicine Based on CiteSpace

[Objectives]This study was conducted to explore the application and development trend of Chinese medicinal material cinnamon in the field of traditional Chinese medicine.[Methods]Articles published from 2008 to 2023 were exported using"cinnamon"as the subject word in the Chinese database of CNKI.Knowledge graphs were drawn by CiteSpace software on the number of articles published,the institutions publishing articles,and keyword clustering,and the data were sorted by Excel.Combined with the extracted information,the application of cinnamon in traditional Chinese medicine and integrated traditional Chinese and Western medicine was analyzed,and its development trend was discussed and prospected,providing further reference for researchers.[Results]The number of articles published showed an overall upward trend and maintained a high number of articles.In the analysis of the journals publishing articles,the journal with the largest number of articles was West China Journal of Pharmaceutical Sciences,which had certain representativeness.In the analysis of institutions publishing articles,the institution with the largest number of articles was Beijing University of Chinese Medicine,and most institutions had little cooperation.Four categories was obtained in keyword clustering,respectively,general research,component identification,production process and product development of cinnamon."Glycyrrhizin"was the keyword with the earliest burst time,and the hot words that have received much attention in recent years are"medication law"and"data mining".[Conclusions]The application of cinnamon in the field of traditional Chinese medicine is mainly to treat diseases and as raw materials for traditional Chinese medicine products.The development trend is"quality control"and"product research and development".Further research and development of cinnamon in traditional Chinese medicine need to promote the participation of more institutions to participate,and cooperation and communication between institutions should be strengthened to promote the deep integration of production,research and academia.

Exploration of Teaching Reform of TCM Dispensing Technology in Higher Vocational Colleges Based on Traditional Chinese Medicine Skills Competition

This paper aims to thoroughly explore the teaching reform of Traditional Chinese Medicine(TCM)dispensing technology in higher vocational colleges,with a focus on the Traditional Chinese Medicine Skills Competition.By analyzing the current teaching status and main challenges of TCM dispensing technology,and aligning with the requirements of the skills competition,a series of detailed and practical teaching reform measures are proposed.These measures are designed to comprehensively improve the teaching quality of TCM dispensing technology in higher vocational colleges,enhance students’practical skills and professional qualities,and ultimately meet the demand for high-quality,skilled talents in the field of TCM.

Progress of Traditional Chinese Medicine Nursing Technology in Rectus Abdominis Diastasis

Rectus abdominis diastasis is a common condition in postpartum women,causing back pain,and in severe cases,it can result in abdominal wall hernias and downward migration of abdominal organs.At present,there is no clearly established optimal non-surgical treatment for rectus abdominis diastasis,but traditional Chinese medicine(TCM)nursing techniques have proven to be an effective intervention for postpartum rectus abdominis separation.This paper comprehensively analyzes the TCM nursing technology as an intervention for postpartum rectus abdominis separation,discusses the shortcomings and prospects of TCM nursing technology,and provides references for further exploring and optimizing the intervention program for postpartum rectus abdominis separation.

Application and Practice of Traditional Chinese Medicine Characteristic Technology Inheritance in Research and Development of Traditional Chinese Medicine Preparations in Medical Institutions

Medical institution preparations play an important role in the development of hospitals,being one of the main forms of traditional Chinese medicine clinical characteristic medication,as well as an important carrier for the inheritance and innovation of traditional Chinese medicine.In recent years,the research and development of traditional Chinese medicine preparations has become a hot spot in the research and development of preparations by medical institutions.Traditional Chinese medicine preparations prepared by traditional processes and their entrusted preparation matters have changed from the current licensing management to filing management.The registration model of traditional Chinese medicine preparations has undergone significant changes,emphasizing more on real data and evaluation results from the objective world,respecting traditional medication experience,and emphasizing the inheritance and innovative development of traditional technological experience.In the process of following teachers in the inheritance project of traditional Chinese medicine,we combined the inheritance of traditional Chinese medicine characteristic technology with the research and development practice of traditional Chinese medicine preparations in medical institutions,and focused on the analysis and discussion of technical problems such as prescription screening,dosage form selection,process route design and clinical observation of traditional Chinese medicine preparations,hoping to provide reference for scholars in the same field of traditional Chinese medicine.

Insights into Fermentation Technology for Traditional Chinese Medicine:Progress and Applications

Traditional Chinese medicine(TCM)has an exemplary role in the treatment and prevention of diseases.However,the advancement of TCM has been constrained by several factors,including its intricate structure,low active ingredient concentration,prolonged growth cycle,and the dfficulty in artificial cultivation.In recent years,research on the fermentation technology of TCM has increased.This paper provides an overview of the advantages of TCM fermentation technology,including efficiency enhancement and toxicity reduction,the development of new drugs,and the secondary utilization of dregs.Additionally,it discusses the progress of research on the application of fermented TCM in animal husbandry and disease treatment.The aim is to provide theoretical guidance for TCM fermentation technology research and the development of fermented TCM.

A systematic review of the efficacy of traditional Chinese medicine retention enema in the treatment of ulcerative colitis

The objective of this study is to summarize the efficacy and safety of traditional Chinese medicine(TCM)enema in the treatment of ulcerative colitis(UC).The randomized controlled trials on TCM enema intervention in the treatment of UC were searched in seven databases:PubMed,Embase,Web of Science,CBM,CNKI,Wanfang Database,and VIP Database from January 1,2013 to June 6,2022,and the data were analyzed using RevMan 5.3 software.A total of 18 studies involving 1514 UC patients were included.Meta analysis results showed that compared with conventional Western medicine,Chinese medicine enema had a significant effect on UC,and the clinical effective rate of the experimental group using Chinese medicine enema was 4.45 times that of the control group using conventional Western medicine(odds ratio=4.45,95%confidence interval[3.27,6.06]).Therefore,Chinese medicine enema is effective in the treatment of UC,and can significantly reduce related symptoms.

Advances in Research Methods of Pharmacodynamic Substances in Traditional Chinese Medicines and Their Application in the Teaching of Traditional Chinese Medicine Analysi

With the continuous progress of science and technology,the research methods of pharmacodynamic substances in traditional Chinese medicine are developing,and the application of these methods in teaching is becoming more and more extensive.By introducing these research methods into the classroom,teachers can help students to deeply understand the nature and mechanism of action of pharmacodynamic substances in traditional Chinese medicine,and improve their interest in and knowledge of traditional Chinese medicine.This paper introduces the definition of pharmacodynamic substances in traditional Chinese medicine,research methods,and their application in the teaching of traditional Chinese medicine analysis.

Traditional Chinese Medicine in colorectal cancer treatment:a bibliometrics study and visualization analysis via CiteSpace

Objective:To evaluate the current state of research and areas of interest for traditional Chinese medicine(TCM)in the field of colorectal cancer treatment.Methods:Related papers published between January 1,2012,and November 27,2021,were found using the Web of Science Core Collection Science Citation Index Expanded.Using CiteSpace's network map generation capability,we then determined the top writers,organizations,countries,keywords,co-cited writers,journals,references,and research trends.Results:This investigation yielded a total of 336 relevant papers.China is the most productive country.Shanghai University of Traditional Chinese Medicine is the leading institution.The journal with the most popularity and publishing volume is Evidence-based Complementary and Alternative Medicine.The author with the most citations and centrality is Lin JM.The terms"epithelial-mesenchymal transition,""cell cycle arrest,""apoptosis,"and"autophagy"are highly frequent and have a high betweenness centrality.Conclusion:According to the results,research on natural products,traditional Chinese medicine(TCM)extracts,and the molecular mechanisms of TCM chemical constituents constitutes the primary focus within TCM cancer treatment investigations.In recent years,there has been a surge of interest in exploring the role of gut microbiota in TCM chemical constituents research,particularly in its ability to induce apoptosis and autophagy in tumor cells,thereby suppressing tumor cell proliferation,metastasis,and invasion.However,due to the intricate composition of TCM and existing technical limitations,the underlying principles guiding TCM's efficacy in treating colorectal cancer remain unclear and warrant further investigation.

Research on Technology Development of Traditional Chinese Medicine Extraction Based on Patent Information

Objective To understand the development status of traditional Chinese medicine extraction technology,so as to provide a reference for enterprises and scientific research institutions to determine the direction of technological innovation.Besides,the government can formulate scientific and technological innovation strategies.Methods Based on patent information,statistical methods were applied to analyze the development of traditional Chinese medicine extraction technology.Then,the core technology was identified by constructing a patent citation network.Finally,the main path analysis method was applied to discover the technological evolution path.Results and Conclusion The extraction technology of traditional Chinese medicine is still in the growth stage.At present,its R&D is based on enterprises,supplemented by scientific research institutions.The core technologies are ultrasonic extraction and enzymatic extraction.The main path represented by enzymatic extraction technology reveals the evolution process of traditional Chinese medicine extraction technology.Based on this,relevant suggestions are put forward.Firstly,technological innovation cooperation should be strengthened to explore new models of achievement transformation and cooperation.Secondly,key enterprises of technology clusters should be cultivated to accelerate breakthroughs in core technologies.Thirdly,combined technologies can achieve high-efficiency enzyme-assisted extraction and open up new ways of extracting traditional Chinese medicine.Fourthly,traditional techniques should be improved to develop innovative modern techniques.

Visual Analysis of Knowledge Map of Traditional Chinese Medicine in Prevention and Treatment of Pulmonary Interstitial Fibrosis Based on CiteSpace

Objective: To analyze the relevant research literature on the prevention and treatment of pulmonary interstitial fibrosis with traditional Chinese medicine (TCM), understand the current research status, hot spots and future development trend in this field, and provide basis and feasible suggestions for further research in this field. Methods: The journal literatures related to the prevention and treatment of pulmonary interstitial fibrosis with TCM in recent 20 years in CNKI database were searched and passed through CiteSpace 5.8.R3 generates the knowledge map of relevant literature authors, document issuing institutions and keywords, and makes visual analysis. Results: A total of 1,576 documents were included, and the annual number of documents showed a fluctuating upward trend, forming a relatively stable research team represented by authors such as LYU Xiaodong, PANG Lijian and LIU Chuang;According to the atlas of document issuing institutions, Shandong University of Traditional Chinese Medicine and its affiliated hospitals ranked first in the number of documents issued, and the cooperation between institutions is dominated by the University of traditional Chinese medicine and its affiliated hospitals;Keyword cluster analysis shows that a large number of studies have been carried out in the field of etiology and pathogenesis, TCM compound, clinic and experiment. Conclusion: The research on the prevention and treatment of pulmonary interstitial fibrosis with TCM has a high degree of attention, but the cooperation network between the research authors and institutions needs to be strengthened. The research on the pathogenesis and improving the quality of life of patients is the trend of development in the future.

Bibliometrics and visual analysis for clinical research on intervention of traditional Chinese medicine nursing technology on rheumatoid arthritis

Objective:The objective of this study is to explore the current situation,clinical research hot spots,and trends of traditional Chinese medicine(TCM)nursing technology in rheumatoid arthritis(RA),to draw a panorama of clinical research,to provide the basis and clues for subsequent high-level evidence integration and further in-depth research.Materials and Methods:Seven databases including China National Knowledge Infrastructure,Chinese Scientific Journal Database(VIP),Wanfang Database,Chinese Biomedical Literature Database(Sino-Med),PubMed,Embase,and the Cochrane Library were searched.The bibliometric method and visualization software CiteSpace were used to conduct a multi-dimensional analysis of the included literature.Results:A total of 805 pieces of literature were included(of them,one was written in German and four in English),and the number of published literature showed an increasing trend year by year.There were only 30(3.73%)pieces of literature published in nursing journals.The hot spots of the co-occurrence map were concentrated in:(1)the nursing of fumigation in RA;(2)the efficacy evaluation of TCM nursing technology on pain,joint malformation,and joint dysfunction caused by RA.The included literature themes focused on five TCM technologies:fumigation,moxibustion,acupoint patching,acupoint injection,and Chinese herbal soaking.The beginning year and strength in TCM fumigation were 2012 and 5.2,and those in moxibustion were 2011 and 3.38.Conclusion:The related studies are on the rise.It has entered the field of international readers with clear research hot spots.However,there are still shortcomings such as little literature published in Chinese nursing core journals,few non-Chinese-related documents,and a lack of international exchanges and cooperation.The current research hot spots in this field are TCM fumigation,and the cutting-edge trend of future research may be fumigation and moxibustion technology.It is suggested that further research can focus on evidence integration and original research on the self-optimization of these two techniques.

Drug Sensitive Test on Main Pathogenic Bacteria of Cow Recessiveness Mastitis Using Traditional Chinese Medicine Preparation “Zengrujianniusan”

According to the theory of Chinese veterinary medicine and characteristics of Chinese herbal medicine, four different preparations of compound Chinese medicine ＂Zengrujianniusan＂ were composed, and the prepared water decoction was used to carry on the bacteriostatic test on main pathogens of cow recessiveness mastitis. The results showed that the four different prescriptions of water decoction all had antibacterial effects. The prescription 3 was sensitive to Staphylococcus aureus and Streptococcus agalactiae, while the other three prescriptions showed high sensitivity, and the prescription 3 had the strongest bacteriostatic effects.

Applications of HPLC/MS in the analysis of traditional Chinese medicines

In China, traditional Chinese medicines （TCMs） have been used in clinical applications for thousands of years. The successful hyphenation of high-performance liquid chromatography （HPLC） and mass spectrometry （MS） has been applied widely in TCMs and biological samples analysis. Undoubtedly, HPLC/MS technique has facilitated the understanding of the treatment mechanism of TCMs. We reviewed more than 350 published papers within the last 5 years on HPLC/MS in the analysis of TCMs. The present review focused on the applications of HPLC/MS in the component analysis, metabolites analysis, and pharmacokinetics of TCMs etc. 50% of the literature is related to the component analysis of TCMs, which show that this field is the most popular type of research. In the metabolites analysis, HPLC coupled with electrospray ionization quadrupole time-of-flight tandem mass spectrometry has been demonstrated to be the powerful tool for the characterization of structural features and fragmentation behavior patterns. This paper presented a brief overview of the applications of HPLC/MS in the analysis of TCMs. HPLC/MS in the fingerprint analysis is reviewed elsewhere.

Construction of Inorganic Elemental Fingerprint and Multivariate Statistical Analysis of Marine Traditional Chinese Medicine Meretricis concha from Rushan Bay

Meretricis concha is a kind of marine traditional Chinese medicine(TCM), and has been commonly used for the treatment of asthma and scald burns. In order to investigate the relationship between the inorganic elemental fingerprint and the geographical origin identification of Meretricis concha, the elemental contents of M. concha from five sampling points in Rushan Bay have been determined by means of inductively coupled plasma optical emission spectrometry(ICP-OES). Based on the contents of 14 inorganic elements(Al, As, Cd, Co, Cr, Cu, Fe, Hg, Mn, Mo, Ni, Pb, Se, and Zn), the inorganic elemental fingerprint which well reflects the elemental characteristics was constructed. All the data from the five sampling points were discriminated with accuracy through hierarchical cluster analysis(HCA) and principle component analysis(PCA), indicating that a four-factor model which could explain approximately 80% of the detection data was established, and the elements Al, As, Cd, Cu, Ni and Pb could be viewed as the characteristic elements. This investigation suggests that the inorganic elemental fingerprint combined with multivariate statistical analysis is a promising method for verifying the geographical origin of M. concha, and this strategy should be valuable for the authenticity discrimination of some marine TCM.

Inorganic Elemental Determinations of Marine Traditional Chinese Medicine Meretricis concha from Jiaozhou Bay:The construction of Inorganic Elemental Fingerprint Based on Chemometric Analysis

The goal of this paper is to explore the relationship between the inorganic elemental fingerprint and the geographical origin identification ofMeretricis concha, which is a commonly used marine traditional Chinese medicine （TCM） for the treatment of asthma and scald bums. For that, the inorganic elemental contents ofMeretricis concha from five sampling points in Jiaozhou Bay have been determined by means of inductively coupled plasma optical emission spectrometry, and the comparative investigations based on the contents of 14 inorganic elements （A1, As, Cd, Co, Cr, Cu, Fe, Hg, Mn, Mo, Ni, Pb, Se and Zn） of the samples from Jiaozhou Bay and the previous reported Rushan Bay were performed. It has been found that the samples from the two bays are ap- proximately classified into two kinds using hierarchical cluster analysis, and a four-factor model based on principle component analysis could explain approximately 75% of the detection data, also linear discriminant analysis can be used to develop a prediction model to distinguish the samples from Jiaozhou Bay and Rushan Bay with accuracy of about 93%. The results of the present investi- gation suggested that the inorganic elemental fingerprint based on the combination of the measured elemental content and chemom- etric analysis is a promising approach for verifying the geographical origin ofMeretricis concha, and this strategy should be valuable for the authenticity discrimination of some marine TCM.

Analysis on the principles of differentiation and prescription of traditional Chinese medicine in the treatment of cancerous fever

To investigate the principles of differentiation and prescription of traditional Chinese medicine(TCM)treatment for cancerous fever by logistic stepwise regression and literature investigation,the following databases were searched(from inception to June 30th 2016):CNKI,CSPD,CBM,PubMed,MEDLINE,EMBASE.According to the inclusion and exclusion criteria,literatures about TCM in the treatment of cancerous fever were screened.The frequency about cancerous fever of differential syndrome types,formulas and herbs were analyzed by logistic multiple regression analysis.The model about syndrome types and formula application of cancerous fever was established.The result showed formula prescriptions used most frequently were Qing-Hao-Bie-Jia decoction,Xiao-Chai-Hu decoction,Xue-Fu-Zhu-Yu decoction,Bai-Hu decoction,San-Ren decoction.The main syndrome types of cancerous fever were as follows:Toxic heat flourishing type,Yin deficiency type,Stagnation of damp-heat type,Qi-Yin deficiency type,Liver meridian depressed-heat type,Blood stasis type and Yang deficiency type.Based on regression equation of the syndrome types,the corresponding herbs of this seven syndrome types were Radix Bupleuri,Radix Paeoniae Alba and Scutellaria Baicalensis Georgi;Radix Bupleuri and Cortex Moutan;Radix Paeoniae Rubra and Radix Angelicae Sinensis;Rhizoma Pinelliae,Cortex Lycii Radicis and Radix Paeoniae Alba;Herba Artemisiae Annuae,Gypsum Fibrosum and Poria Cocos;Rhizoma Anemarrhenae,Scutellaria Baicalensis Georgi and Radix Codonopsis;Radix Bupleuri respectively.The results of Logistic regression analysis and frequency of herbs statistics can provide reference for differentiation and treatment of cancerous fever in clinical.

Visual analysis of research hotspots and trends in the treatment of immune thrombocytopenia with traditional Chinese medicine

Objective Through bibliometrics and visual analysis of the related studies on traditional Chinese medicine(TCM)treatment of immune thrombocytopenia(ITP),this study aims to sort out the overall research progress,hotspots,and trends in this field,and provide reference for further research in ITP.Methods The articles on ITP treated by TCM were retrieved from China National Knowledge Infrastructure(CNKI),Wanfang Database,China Science and Technology Journal Database(VIP),Web of Science Core Collection(WOSCC),and PubMed.The retrieval time was from the establishment of the databases to July 31,2022.VOSviewer,CiteSpace,Carrot2,and Note-Express were used for data analysis of the articles in terms of their quantities,types,and journals,and for visualization of research hotspots,authors,institutions,and keywords.Results 1493 Chinese articles and 40 English articles were included.The articles in Chinese mainly focus on clinical trial research and clinical experience summary,while the English articles mainly focus on clinical trial research and animal research.The Chinese articles were published in 317 Chinese journals,while English articles were published in 29 English journals.Research hotspots include the clinical syndrome differentiation of ITP,the therapeutic effect of TCM compounds on ITP,and the mechanism of ITP treatment.Keyword analysis shows that there are many research achievements in integrated traditional Chinese and western medicine treatment,clinical research,famous doctors’experience,TCM treatment,cellular immunity,and humoral immunity.The authors with the most articles in Chinese and English are Professor CHEN Xinyi and Professor MA Rou,respectively,and the research institutions with the most articles are Dongzhimen Hospital of Beijing University of Chinese Medicine and Xiyuan Hospital of China Academy of Chinese Medical Sciences.Chinese herbs often used to treat ITP clinically include Xianhecao(Agrimoniae Herba),Nvzhenzi(Ligustri Lucidi Fructus),Mohanlian(Ecliptae Herba),Zhongjiefeng(Sarcandrae Herba),etc.,and the prescription usually used to treat ITP include Guipi Decoction(归脾汤),Xijiao Dihuang Decoction(犀角地黄汤),Bazhen Decoction(八珍汤),Erzhi Pill(二至丸),and Xiaochaihu De-coction(小柴胡汤).The main development trends toward retrospective study,TCM treatment mechanism,and data mining.

Prescription analysis of traditional chinese medicine for COVID-19treatment

Background:Coronavirus disease 2019 is still spreading all over the world and without any specific medicine.Traditional Chinese medicine has played an important role in the control of epidemics in China and accumulated many experiences.The aim of this study is to summarize the law of traditional Chinese medicine and provide reference for coronavirus disease 2019 traditional Chinese medicine treatment.Methods:We collected the treatment plans of traditional Chinese medicine for coronavirus disease 2019 in 24 provinces of China to evaluate the basic Chinese medicine formulas,and the efficacy types,frequency,“Four Characters”,“Five Tastes”,“Channel Tropism”of herbal medicines.Combination rules between medicines were calculated by Apriori algorithm and Clementine12.0.Results:The formulas for“Relieving Exterior Syndrome”,“Expelling Dampness”,“Clearing Heat”and“Supplementing”have highest proportion.In prevention period and medical observation period,the medicines mainly have effect for“Clearing Heat”,“Relieving Exterior Syndrome”and“Supplementing”.In treatment period,medicines for“Clearing Heat”,“Relieving Superficies”and“Relieving Dampness”had higher frequency.The“Four Characters”of Chinese medicines in all treatment plans were mainly“Cold”and“Warm”.Medicines in the treatment plans always belong to“Pungent Taste”,“Bitter Taste”and“Sweet Taste”.For“Channel Tropism”,most medicines turned over to the“Lung”,“Stoma”and“Spleen”.The common medicine combinations in the recommended formulas during the prevention period and medical observation period were medicines for“Qi-Supplementing”and“Relieving Superficies”.In treatment period(mild,moderate,severe/critical),medicines for“Dampness Expelling”,“Relieving Superficies”and“Qi-Supplementing”were combined in a high frequency.In recovery period,the medicines for“Dispelling Dampness”combined the medicines for“Promoting Blood Circulation”with highest frequency.Conclusions:We found the characteristics of medicine and prescription rules of traditional Chinese medicine for the treatment of coronavirus disease 2019 and provided scientificity and effectiveness of it.More importantly,it provides reference for the development of new clinical prescriptions for coronavirus disease 2019.

Screening and Efficacy Observation of Traditional Chinese Medicine Against Chicken Pathogenic Escherichia coli

[ Objective] The aim of this study was to lay a foundation for obtaining effective traditional Chinese medicine preparations controlling chicken colibacillosis. [ Method] With the western medicine Lomeflbxacin Hydrochloride and Sodium Chloride Injection as the control, drug sensitivity tests of 15 single traditional Chinese herbs and seven self-made Chinese herbal compounds were conducted. Subsequently, the single traditional Chinese herbs presenting good in vitro antibacterial effects against Escherichia coli and all the seven compounds were given to the chicks artificially infected with E. coil to observe their efficacies. [ Result] The results of drug sensitivity tests showed that all the 15 traditional Chinese herbs had certain antibacterial effect against E. coli, among which aloe, pomegranate rind and radix sanguisorbae were the best. Compound 1, 2 and 5 had good antibacterial effects in vitro. By in vivo test, traditional Chinese medicines showed superior efficacies on infected chicks than western medicine, and compound 1 and 2 performed well. [ConclusiOn] Traditional Chinese medicine preparations are good at controlling Chicken colibacillosis, but their bacteriostasis mechanism in vivo was different with that in vitro.