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PubMed2002

DNA fingerprinting of sister blastomeres from human IVF embryos.

BACKGROUND: Previously published single cell DNA fingerprinting systems have been plagued by high rates of allele drop-out (ADO) and preferential amplification (PA) preventing clinical application in preimplantation genetic diagnosis. METHODS: Tetranucleotide microsatellite markers with high heterozygosity, known allelic size ranges and minimal PCR stutter artefacts were selected for chromosomes X, 13, 18 and 21 and optimized in a multiplex fluorescent (FL)-PCR format. FL-PCR products were analysed using the ABI Prism 377 DNA sequenator and Genescan software. Validation of the DNA fingerprinting system was performed on single diploid (n = 50) and aneuploid (n = 25) buccal cells and embryonic blastomeres (n = 21). RESULTS: The optimized pentaplex PCR DNA fingerprinting system displayed a high proportion of successful amplifications (>91%) and low ADO and PA (<6%) when assessed on 50 human buccal cells. DNA fingerprints of single cells from a subject with Down's syndrome detected the expected tri-allelic pattern for the chromosome 21 marker, confirming trisomy 21. In a blind study on 21 single blastomeres, all embryos were identifiable by their unique DNA fingerprints and shared parental alleles. CONCLUSIONS: A highly specific multiplex FL-PCR based on the amplification of five highly polymorphic microsatellite markers was developed for single cells. This finding paves the way for the development of a more complex PCR DNA fingerprinting system to assess aneuploidy and single gene mutations in IVF embryos from couples at genetic risk.

Alleles↗

PubMedDate not supplied

DNA fingerprinting confirms isogenicity of androgenetically derived rainbow trout lines.

Homozygous and hybrid clonal lines of rainbow trout (Oncorhynchus mykiss) were confirmed to be isogenic using multilocus DNA fingerprinting. Homozygous clones were produced by androgenesis and gynogenesis using gametes from androgenetic male and female rainbow trout, respectively. Isogenic F1 hybrid lines were produced by crossing homozygous fish from different strains. One line of hybrid clones showed segregation for maternally inherited DNA fingerprint markers. The female from this cross, the only presumptive homozygous gynogenetic individual used in this study, was thought to have been produced by gynogenesis followed by blockage of the first cleavage division, but based on the DNA fingerprint analysis, apparently was derived by spontaneous polar body retention that maintained heterozygosity at some loci. Mutations at DNA fingerprint loci were not observed, indicating relative stability of fingerprint loci in the clonal lines. DNA fingerprinting appears to be a useful tool for identifying and genetically monitoring clonal lines of rainbow trout. Isogenic lines of rainbow trout will facilitate the production of saturated genetic maps for rainbow trout and enhance such endeavors as quantitative trait locus (QTL) analysis and loss of heterozygosity (LOH) studies in tumors.

Androgens↗

PubMed2000

Contigs built with fingerprints, markers, and FPC V4.7.

Contigs have been assembled, and over 2800 clones selected for sequencing for human chromosomes 9, 10 and 13. Using the FPC (FingerPrinted Contig) software, the contigs are assembled with markers and complete digest fingerprints, and the contigs are ordered and localised by a global framework. Publicly available resources have been used, such as, the 1998 International Gene Map for the framework and the GSC Human BAC fingerprint database for the majority of the fingerprints. Additional markers and fingerprints are generated in-house to supplement this data. To support the scale up of building maps, FPC V4.7 has been extended to use markers with the fingerprints for assembly of contigs, new clones and markers can be automatically added to existing contigs, and poorly assembled contigs are marked accordingly. To test the automatic assembly, a simulated complete digest of 110 Mb of concatenated human sequence was used to create datasets with varying coverage, length of clones, and types of error. When no error was introduced and a tolerance of 7 was used in assembly, the largest contig with no false positive overlaps has 9534 clones with 37 out-of-order clones, that is, the starting coordinates of adjacent clones are in the wrong order. This paper describes the new features in FPC, the scenario for building the maps of chromosomes 9, 10 and 13, and the results from the simulation.

Chromosomes, Human, Pair 10↗

PubMed1995

Diversity of DNA fingerprints in Cryptococcus neoformans.

DNA fingerprint patterns of 156 Cryptococcus neoformans isolates (26 AIDS patients, 46 non-AIDS patients, and 40 environmental sources) from both varieties (126 C. neoformans var. neoformans and 30 C. neoformans var. gattii isolates) and from seven countries were analyzed by using the DNA probe UT-4p. Nine and twelve distinct DNA fingerprint patterns were observed for isolates of the C. neoformans var. neoformans and var. gattii, respectively. No pattern was unique to AIDS patients, non-AIDS patients, or the environment. Pattern II was observed more often in non-AIDS patients (8 of 23) than in AIDS patients (0 of 25). Pattern V was the most prevalent pattern (42 of 82) in clinical and environmental isolates. Isolates from three AIDS patients in Burundi and Zaire exhibited patterns identical to each other but different from those of isolates collected from their houses (i.e., dust of floors, walls, etc.) or a nearby pigeon coop. DNA fingerprint stability was determined for 53 isolates from nine non-AIDS patients at different time intervals during 5 to 128 weeks of antifungal therapy. For eight patients, the fingerprint pattern was stable while the ninth may have had a mixed infection. Pattern II was observed in 4 of 9 patients, which is similar to 4 of 14 in other non-AIDS patients as reported here. In spite of the extensive pattern heterogeneity among 15 C. neoformans var. gattii isolates in Australia, the patterns observed in seven California isolates were quite different from those in Australia. Among isolates of C. neoformans var. gattii, one fingerprint pattern (designated b) was observed in several countries of the Far East. The fingerprint patterns of two of three environmental isolates from Eucalyptus camaldulensis trees in Australia were identical to those of 2 of the 12 clinical isolates from the country.

AIDS-Related Opportunistic Infections↗

PubMed1997

IS6110 fingerprinting of drug-resistant Mycobacterium tuberculosis strains isolated in Germany during 1995.

The epidemiological relatedness of drug-resistant Mycobacterium tuberculosis strains isolated in Germany in 1995 was evaluated by the standardized IS6110 fingerprinting method. Altogether, 196 M. tuberculosis isolates from 167 patients were analyzed. A large degree of IS6110 polymorphism was found, ranging from 1 to 20 copies. Multiple isolates from one patient generally remained stable over a period of up to 1 year. However, one strain showed an additional fragment 7 months after the first isolate was obtained. Isolates from 55 patients (33%) showed identical fingerprint patterns or fingerprint patterns that differed only in one band, and thus they were clustered in 22 fingerprint groups. Specific transmission links could be established between members of four groups, e.g., transmission by family contacts. In one case, transmission of a multidrug-resistant strain to a patient initially infected with a drug-susceptible strain could be shown. Besides these fingerprint groups, 30 of the 167 isolates (approximately 18%) could be grouped in two fingerprint clusters with a similarity of at least 78%. Approximately 60% of the patients of these two clusters were known to be immigrants from the former Soviet Union, and one patient is still living in Belarus. In conclusion, our results indicate that (i) transmission of drug-resistant strains contributes substantially to the emergence of drug-resistant tuberculosis in Germany and (ii) drug-resistant M. tuberculosis strains were presumably carried over from the former Soviet Union to Germany by immigrants.

Commonwealth of Independent States↗

PubMed1997

Identification of Candida species by randomly amplified polymorphic DNA fingerprinting of colony lysates.

We have characterized a method that produces simple yet diagnostic fingerprints that are unique to isolates of Candida species. DNA from individual colonies can be amplified from crude single-colony lysates. Randomly amplified polymorphic DNA (RAPD) fingerprints generated from a single primer correctly identified the species of most (>98%) of the isolates identified with CHROMagar Candida plates as non-Candida albicans Candida species. RAPD fingerprints were much more informative than the plates, since they distinguished between all tested species and required less time. Most (91%) of these identifications agreed with those assigned by API 20C tests. In almost every incident of species identity mismatch, electrophoretic karyotyping showed that the RAPD fingerprint was correct. This underscores the improved objectivity and reliability of this method over those of conventional diagnostic tools. The identities of approximately 30% of C. albicans isolates identified in clinical laboratories by positive germ tube tests are not verified by either testing on CHROMagar Candida plates or RAPD fingerprinting. Data suggest that clinical isolates conventionally identified as C. albicans in clinical settings are heterogeneous, consisting of both misidentified and atypical yeasts. RAPD fingerprints obtained from primary culture plate colonies allows for rapid, highly accurate determinations of Candida species, hence permitting earlier selection of appropriate antifungal agents in the clinical setting.

Candida↗

PubMed2003

DNA fingerprinting of Lactobacillus crispatus strain CTV-05 by repetitive element sequence-based PCR analysis in a pilot study of vaginal colonization.

Lactobacillus crispatus is one of the predominant hydrogen peroxide (H(2)O(2))-producing species found in the vagina and is under development as a probiotic for the treatment of bacterial vaginosis. In this study, we assessed whether DNA fingerprinting by repetitive element sequence-based PCR (rep-PCR) can be used to distinguish the capsule strain of L. crispatus (CTV-05) from other endogenous strains as well as other species of vaginal lactobacilli. Vaginal and rectal lactobacilli were identified to the species level by using whole-chromosome probe DNA hybridization. The DNAs from L. crispatus, L. jensenii, L. gasseri, and an as-yet-unnamed H(2)O(2)-negative Lactobacillus species designated 1086V were subjected to rep-PCR. The results of gel electrophoresis and ethidium bromide staining of the DNA fingerprints obtained were compared. L. crispatus CTV-05 had a unique DNA fingerprint compared to all other lactobacilli. DNA fingerprints for 27 production lots of L. crispatus sampled from 1994 through 2001 were identical to that of the original strain isolated in 1993, suggesting strain stability. In a pilot study of nine women, this DNA fingerprinting method distinguished CTV-05 from other endogenous vaginal lactobacilli prior to and after vaginal capsule use. rep-PCR DNA fingerprinting is useful for strain typing and for evaluating longitudinal loss or acquisition of vaginal lactobacilli used as probiotics.

Bacterial Typing Techniques↗

PubMed1993

Characterization of Actinomyces with genomic DNA fingerprints and rRNA gene probes.

Cellular DNA from 25 Actinomyces naeslundii and Actinomyces viscosus strains belonging to the 7 taxonomic clusters of Fillery et al. (1978) and several unclustered strains was obtained by enzymatic and N-lauroylsarcosine/guanidine isothiocyanate treatment of whole cells, followed by extraction of the nucleic acid. The DNA samples were digested with restriction endonucleases BamHI or PvuII, and agarose gel electrophoresis was used to obtain DNA fingerprints. The DNA fragments were subjected to Southern blot hybridization with a digoxigenin-labeled cDNA probe transcribed from Escherichia coli 16S and 23S rRNA. The patterns of bands from genomic (DNA fingerprints) and rDNA fingerprints (ribotypes) were used for comparison between the taxonomic cluster strains and strains within clusters. Representative strains from each taxonomic cluster provided different BamHI DNA fingerprints and ribotype patterns with 3 to 9 distinct bands. Some strains within a cluster showed identical ribotype patterns with both endonucleases (A. naeslundii B120 and A. naeslundii B102 from cluster 3), while others showed the same pattern with BamHI but a different pattern with PvuII (A. naeslundii ATCC 12104 and 398A from cluster 5). A viscosus ATCC 15987 (cluster 7) and its parent strain T6 yielded identical fingerprint and ribotype patterns. The genomic diversity revealed by DNA fingerprinting and ribotyping demonstrates that these techniques, which do not require phenotypic expression, are suited for study of the oral ecology of the Actinomyces, and for epidemiological tracking of specific Actinomyces strains associated with caries lesions and sites of periodontal destruction.

Actinomyces↗

PubMed1995

Application of DNA fingerprinting with digoxigenated oligonucleotide probe (CAC)5 to analysis of the genetic variation within Taenia taeniaeformis.

DNA from T. taeniaeformis digested with the restriction endonuclease was hybridized with digoxigenated oligonucleotide probe (CAC)5. Metacestode and adult showed same clear multibanding patterns, which were characteristic of multilocus DNA fingerprinting. The fingerprinting patterns were quite different from those of the rodent hosts. Genetic variations in 4 laboratory-reared isolates of T. taeniaeformis, including 3 isolates which have been reported to be indistinguishable by infectivity, morphology and protein composition of metacestode, were investigated using this technique. Each of the 4 isolates exhibited isolate-specific fingerprinting patterns and were easily distinguished from one another, thus it was considered that (CAC)5 was a highly resolvable and informative probe for cestodes. However, it was also indicated that (CAC)5 was so sensitive that applying fingerprinting with (CAC)5 to taxonomical or phylogenetic analysis was limited where habitat of the host was restricted to the small area. In comparison to fingerprinting with 32P-labeled (CAC)5, fingerprinting with digoxigenated (CAC)5 represented more and sharper bands. It was considered that a digoxigenated probe was more useful for genetic analysis of cestodes.

Animals↗

PubMed1994

Identification of sublines of inbred strains of mice and assessment of genetic relationships between substrains or sublines by DNA fingerprinting.

Identification of substrains or sublines of inbred mice and assessment of genetic relationships among them were performed on the basis of DNA fingerprinting using M13 phage DNA as a probe. We used eight C57BL/6 sublines (J parallel Jcl, J parallel Jms, J parallel Slc, J parallel Nrs, J parallel Yok, Jah, N parallel Crj, N parallel Jcl) and eleven C3H/He sublines, (J, J parallel Jcl, J parallel Yok, J parallel Nrs, J parallel Jms, N, N parallel Jcl, N parallel Crj, Slc, Jah, Nrs). Two kinds of restriction endonucleases (HinfI and PstI) were used. It was found that: 1) DNA fingerprint within each subline showed identical patterns. 2) Most sublines of C57BL/6 and C3H/He could be identified using DNA fingerprinting with HinfI except between N parallel Crj and Slc, and among J, J parallel Nrs and J parallel Yok in C3H/He. DNA fingerprints with PstI endonucleases showed low polymorphic banding patterns. 3) A dendrogram constructed from DNA fingerprint patterns reflected generally the genealogy of the sublines used. 4) DNA fingerprinting, therefore, seemed to be suitable for the genetic monitoring and assessment of genetic relationships among sublines of inbred mice having close relationships.

Animals↗

PubMed2000

Single primer polymerase chain reaction fingerprinting for Pasteurella multocida isolates from laboratory rabbits.

OBJECTIVE: To evaluate a rapid polymerase chain reaction (PCR) fingerprinting technique for discriminating among Pasteurella multocida isolates from laboratory rabbits. SAMPLE POPULATION: 33 P multocida isolates from rabbits with clinical pasteurellosis. PROCEDURE: PCR assays were conducted with 2 minisatellites (core sequence and modified core sequence of phage M13) and 2 microsatellites ([GTG]5 and [GACA]4). Each bacterium was assigned to a PCR type for each of the primers used. Boiled bacterial extracts and purified genomic DNA were compared by use of PCR assays for phage M13 and (GACA)4. Plasmids were isolated from each bacterium, and their influence on PCR fingerprint was determined, using boiled extracts as a DNA source. RESULTS: M13 core sequence and M13 modified core sequence yielded 5 and 8 PCR types, respectively. The microsatellites (GTG)5 and (GACA)4 yielded 4 and 9 PCR fingerprint types, respectively. Fingerprint patterns obtained by use of isolated DNA differed from those obtained by use of boiled extracts, although discrimination among P multocida isolates was similar. The presence or absence of plasmids did not affect PCR fingerprints. CONCLUSION: Single primer PCR fingerprinting with minisatellite and microsatellite primers is an efficient and reproducible method for the discrimination of P multocida isolates from rabbits and can be performed directly, using boiled bacterial extracts as a source of template, although more bands were obtained from pure genomic DNA.

Animals↗

PubMed1995

Cluster analysis of Helicobacter pylori genomic DNA fingerprints suggests gastroduodenal disease-specific associations.

BACKGROUND: Helicobacter pylori infection is now accepted as the most common cause of chronic active gastritis and peptic ulcer disease. The etiologies of many infectious diseases have been attributed to specific or clonal strains of bacterial pathogens. Polymerase chain reaction (PCR) amplification of DNA between repetitive DNA sequences, REP elements (REP-PCR), has been utilized to generate DNA fingerprints to examine similarity among strains within a bacterial species. METHODS: Genomic DNA from H. pylori isolates obtained from 70 individuals (39 duodenal ulcers and 31 simple gastritis) was PCR-amplified using consensus probes to repetitive DNA elements. The H. pylori DNA fingerprints were analyzed for similarity and correlated with disease presentation using the NTSYS-pc computer program. RESULTS: Each H. pylori strain had a distinct DNA fingerprint except for two pairs. Single-colony DNA fingerprints of H. pylori from the same patient were identical, suggesting that each patient harbors a single strain. Computer-assisted cluster analysis of the REP-PCR DNA fingerprints showed two large clusters of isolates, one associated with simple gastritis and the other with duodenal ulcer disease. CONCLUSIONS: Cluster analysis of REP-PCR DNA fingerprints of H. pylori strains suggests that duodenal ulcer isolates, as a group, are more similar to one another and different from gastritis isolates. These results suggest that disease-specific strains may exist.

Adult↗

PubMed2001

Identification of genomic imbalances in gastric MALT lymphoma using arbitrarily primed PCR DNA fingerprinting.

Arbitrarily primed PCR (AP-PCR) is a unique method to identify the cancer cell specific losses and gains of chromosomal regions by targeting specific genes or chromosomal segments. In the present study, introducing the AP-PCR technique with a single primer, we have ascertained the gains and losses of DNA fingerprints in 15 MALT lymphoma samples. Out of 15 prominent DNA fingerprints, the signal intensity of two fingerprints, labeled bands G and I, were significantly lower in 40 and 50% of tumors as compared to adjacent normal DNA fingerprints, respectively. Similarly, gains of signal intensity of DNA fingerprints (bands A and C) were detected in 13% of tumor samples studied. Variations in signal intensities were also found in other bands within a few samples. Although, the functional importance of these bands is unknown, this study indicates that the AP-PCR generated under or over amplified DNA fingerprints may participate during the progression of MALT lymphoma in human stomach. Moreover, these studies also suggest that the AP-PCR technique, with different primers, can be utilized for the determination of new chromosomal segments in MALT lymphoma samples that can be used for the identification of these diseases.

Biopsy↗

PubMed2002

[Assessment of the injunction regarding genetic fingerprinting of sex offense perpetrators].

OBJECTIVE: To assess the problems eventually created by fingerprinting for genetic identification of suspected sexual aggressors. METHOD: An opinion poll was conducted among 13 practitioners (8 forensic experts and 5 biologists) using a list of 20 questions. RESULTS: Ten practitioners agreed that there were ethical problems in fingerprinting, and 6 of the 13 physicians surveyed claimed that fingerprinting was distressful for the suspect. COMMENTS: The ethical problems most frequently encountered were the presence of a third party during fingerprinting, the practitioners' lack of information at to the test's future use, the suspect's refusal to undergo such a test and the suspect's lack of information on the subsequent computerized treatment of the fingerprint, to enhance a national genetic data base for example, and the length of time during which the fingerprint is stored.

Adult↗

PubMed2003

[DNA fingerprinting of Mycobacterium tuberculosis strains from Beijing, Guangdong and Ningxia].

OBJECTIVE: To explore the epidemic distribution of Mycobacterium tuberculosis isolates from Beijing, Guangdong and Ningxia, and to determine M. tuberculosis strains of the "Beijing Family". METHODS: Two hundred and six IS6110 DNA fingerprinting patterns of M. tuberculosis strains from three provinces (city) were transferred to digital data, compared with the world M. tuberculosis DNA fingerprinting database, and then clustered by Gel compare 4.1 software. The clustering values in different patients with tuberculosis were compared by chi(2) test. Risk factors for recent transmission were calculated using odd ratios. RESULTS: No M. tuberculosis strains were found the same as those of DNA fingerprint database. 56.8% (117/206) fingerprinting patterns of M. tuberculosis shared by least two-thirds of the IS6110 fragments and their Spoligotyping fingerprinting patterns were consistent with those of M.tuberculosis strains of the "Beijing Family". There were significant differences between female and male, different age groups (< 42 years old) and older (>or= 42 years old) (P < 0.05). Odd ratio was 5.06 in the group younger than 42 years old (95% CI: 1.00 - 34.34) and was 4.43 (95% CI: 0.94 - 28.76) in males. CONCLUSION: M. tuberculosis strains of "Beijing Family" were popular in Beijing, Guangdong and Ningxia. Men and younger age group (< 42) were shown to be infected by identical strains more often than women and older aged which might play an important role in the recent transmission of tuberculosis in these areas. IS6110 DNA fingerprinting of M. tuberculosis could be used to trace the source of tuberculosis infection.

Adult↗

PubMed1992

Histogenetic analysis of ovarian germ cell tumors by DNA fingerprinting.

The histogenesis of ovarian germ cell tumors (11 mature teratomas, three malignant transformations of mature teratomas, two immature teratomas, and four dysgerminomas) was investigated genetically using minisatellite DNA probes 33.15 and 33.6 for person-specific restriction fragment length polymorphism (DNA fingerprint) analysis. The DNA fingerprints of six ovarian teratomas were identical with those of mononuclear cells from each host, while some polymorphic bands observed in the host mononuclear cells were lost in the DNA fingerprints of the other five cases. The cases of malignant transformation of mature teratoma and immature teratoma showed that some polymorphic bands of DNA fingerprints from the host mononuclear cells were absent in the tumor tissues. In four cases with dysgerminomas, the DNA fingerprints of tumors were completely identical with those of the respective host mononuclear cells. The present results suggest that mature cystic teratomas of the ovary arise from germ cells arrested at various stages of meiosis, while immature teratomas are derived from postmeiotic germ cells. Malignant transformation may occur exclusively in the mature teratomas arising from postmeiotic germ cells. Dysgerminomas develop from premeiotic oogonia (primordial germ cells). Thus, DNA fingerprints are a useful and sensitive tool for identifying the pathogenesis of germ cell tumours.

Adolescent↗

PubMed2006

Fingerprint chromatogram analysis of Pseudostellaria heterophylla (Miq.) Pax root by high performance liquid chromatography.

A simple and reliable high performance liquid chromatographic (HPLC) method has been developed and validated for the fingerprinting of extracts from the root of Pseudostellaria heterophylla (Miq.) Pax. HPLC with gradient elution was performed on an authentic reference standard of powdered P. heterophylla (Miq.) Pax root and 11 plant samples of the root were collected from different geographic locations. The HPLC chromatograms have been standardized through the selection and identification of reference peaks and the normalization of retention times and peak intensities of all the common peaks. The standardized HPLC fingerprints show high stability and reproducibility, and thus can be used effectively for the screening analysis or quality assessment of the root or its derived products. Similarity index calculations based on cosine angle values or correlation methods have been performed on the HPLC fingerprints. As a group, the fingerprints of the P. heterophylla (Miq.) Pax samples studied are highly correlated with closely similar fingerprints. Within the group, the samples can be further divided into subgroups based on hierarchical clustering analysis (HCA). Sample grouping based on HCA coincides nicely with those based on the geographical origins of the samples. The HPLC fingerprinting techniques thus have high potential in authentication or source-tracing types of applications.

Caryophyllaceae↗

PubMed2007

Screening and analysis of the multiple absorbed bioactive components and metabolites of Dangguibuxue decoction by the metabolic fingerprinting technique and liquid chromatography/diode-array detection mass spectrometry.

Based on the metabolic fingerprinting technique and liquid chromatography/diode array detection mass spectrometry (LC/DAD-MS), a method for rapid screening and analysis of the multiple absorbed bioactive components and metabolites of an oral solution of Dangguibuxue decoction (ODD) in rabbit plasma after oral administration of ODD was developed. The results obtained from a comprehensive comparative analysis of the fingerprints of the ODD and its metabolic fingerprints in rabbit plasma indicated that 46 components in the ODD were absorbed into the rabbit's body. Of them, ten components were tentatively identified from their MS and UV spectra and retention behaviors by comparing the results with the reported literature. They were calycosin-7-O-beta-D-glycoside, (6aR,-11aR)-hydroxy-9,10-dimethoxypterocarpan-3-O-beta-D-glycoside, ononin, L-3-hydroxy-9,10-dimethoxypterocarpan, formononetin, (3R)-7,2'-dihydroxy-3',4'-dimethoxyisoflavan, sedanenolide, E-ligustilide, Z-ligustilide, and Z-butylidenephthalide. In addition, 21 components were only found in the metabolic fingerprints, which suggested that they might be metabolites of some components in the ODD. The findings demonstrated that the proposed method could be used to rapidly and simultaneously analyze and screen the multiple absorbed bioactive constituents and metabolites in a formula of traditional Chinese medicines (TCMs) by comparing and contrasting the chromatographic fingerprints with its metabolic fingerprints. This is very important not only for the pharmaceutical discovery process and the quality control of crude drugs, but also to explain the curative mechanism of TCMs.

Administration, Oral↗