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This study aimed to explore the correlation between rhizosphere soil microorganisms of wild Arnebia euchroma and the content of medicinal components to provide guidance for the selection of the ecological planting base. The total DNA of rhizosphere soil microorganisms of wild A. euchroma was extracted, and the microbial community structure of rhizosphere soil microorganisms was analyzed by IlluminaMiseq high-throughput sequencing technology. The content of total hydroxynaphthoquinone pigment and β,β'-dimethylacrylalkannin in medicinal materials was determined by high-performance liquid chromatography(HPLC). The physicochemical pro-perties of rhizosphere soil of wild A. euchroma in main producing areas were determined, and the correlation of soil microbial abundance with index component content and soil physicochemical properties was analyzed by SPSS software. The results showed that the species composition of rhizosphere fungi and bacteria in A. euchroma from different habitats was similar at the phylum and genus levels, but their relative abundance, richness index(Chao1), and community diversity(Simpson) index were different. Correlation analysis showed that the content of available phosphorus in soil was positively correlated with the content of total hydroxynaphthoquinone pigment and β,β'-dimethylacrylalkannin, and the abundance of five fungal genera such as Solicoccozyma and six bacterial genera such as Pseudo-nocardia and Bradyrhizobium was positively correlated with the content of medicinal components in medicinal materials. The abundance of Bradyrhizobium was significantly positively correlated with the content of β,β'-dimethylacrylalkanin. The abundance of fungi such as Archaeorhizomyces was significantly positively correlated with the content of available phosphorus in rhizosphere soil, and Bradyrhizobium was significantly negatively correlated with soil pH. Therefore, the abundance of fungi and bacteria in the rhizosphere of A. euchroma has a certain correlation with the medicinal components and the physicochemical properties of the rhizosphere soil, which can provide a scientific basis for the selection of ecological planting bases in the later stage.
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Rhizosphère , Microbiologie du sol , Bactéries/génétique , Phosphore , Sol , BoraginaceaeRésumé
This study established high-performance liquid chromatography(HPLC) fingerprints of Chinese medicines derived from Apocynum venetum and Poacynum pictum in Xinjiang and explored their composition differences with the combination of content determination, similarity analysis, cluster analysis and principal component analysis. The HPLC conditions included Phenomenex Kinetex C_(18) column(4.6 mm ×100 mm, 2.6 μm), acetonitrile-0.01% trifluoroacetic acid aqueous solution as mobile phase, gradient elution, flow rate of 0.6 mL·min~(-1), detection wavelength of 281 nm and column temperature of 25 ℃. The content of chlorogenic acid, quercetin-3-O-sophoroside, rutin, hyperin, isoquercitrin, trifolin and astragalin was determined in 31 batches of medicinal materials, and fingerprint research and chemometric analysis were performed with Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine(Version 2004 A) and SPSS 21.0. In the Chinese Pharmacopoeia 2020, the quality of Apocyni Veneti Folium is controlled by character identification, microscopic identification, thin layer chromatography identification and quantitative determination of hyperin. There were 21 common peaks of A. venetum and P. pictum in the HPLC fingerprints, 5 of which were identified as chlorogenic acid, hyperin, isoquercitrin, trifolin and astragalin, with their content also determined. Except for 3 batches of medicinal materials, the similarity of other 28 batches was higher than 0.83, indicating good similarity. Two categories were formed in the cluster analysis based on content determination, which showed that some differences existed in similarities between different regions of Xinjiang. The medicinal materials were ranked by quality with principal component analysis, and the results indicated that the top 15 all came from northern Xinjiang. The quality difference of A. venetum and P. pictum had a correlation with the place of origin. This study provides a reference for the analysis and evaluation of A. venetum and P. pictum from different habitats and the selection of introduction and cultivation areas.
Sujets)
Apocynum , Chine , Chromatographie en phase liquide à haute performance , Médicaments issus de plantes chinoises , Médecine traditionnelle chinoiseRésumé
Based on the results of the fourth national survey of traditional Chinese medicine resources in Turpan city, Xinjiang, this study counted the types of traditional Chinese medicine resources in Turpan Basin. The spatial distribution differences of traditional Chinese medicine resources in Turpan Basin of Xinjiang were analyzed by using grid technology, trend surface analysis, global spatial autocorrelation analysis, and local spatial autocorrelation analysis, so as to clarify the overall change trend and aggregation degree of traditional Chinese medicine resources in Turpan Basin in horizontal and vertical directions. The results showed the following: in the horizontal direction, the species richness of traditional Chinese medicine resources in the central part of Turpan Basin was high, and there were great differences in the species richness of traditional Chinese medicine resources in Turpan Basin under different grid sizes. The spatial scale effect of the richness of traditional Chinese medicine resources in Turpan Basin is obvious. Among them, under the 30 km×30 km scale, the richness of the types of Chinese medicine resources shows a high spatial correlation, and the richness of the types of Chinese medicine resources at 5 km×5 km scale presents a near random distribution state, and the richness of the types of Chinese medicine resources at 80, 90, and 100 km scale sits negatively related. Vertical direction, Chinese medicine resources appear rich at the range of-154-150 m and 900-1 050 m following by range of 1 050-1 200 m.
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Chine , Médecine traditionnelle chinoise , Analyse spatiale , TechnologieRésumé
The quality control of traditional Chinese medicine provides the premise of its modernization and globalization. Currently, the dual quality control based on chemical benchmark and effect benchmark has been recognized domestically and internationally. Research efforts have lead to establishment of a series of effective quality control methods based on chemical components, medicinal properties, microscopic characteristics, material constituents and pharmacodynamic targets. In the study of quality control based on chemical benchmarks, fruitful results on fingerprints, DNA barcodes, and quality markers have been achieved. However, due to a variety of factors, such as growth period, origin, growth environment and preparation process of traditional Chinese medicine, the quality control of traditional Chinese medicine based on chemical benchmarks remains difficult to fully reflect the quality of traditional Chinese medicine. At present, there is still a dispute on how to accurately reflect the quality of traditional Chinese medicines based on chemical benchmarks. For example, the index components selected in the Chinese medicine quality standards are difficult to totally reflect all the components of Chinese medicine, and the relevance between the index components versus therapeutic effect is not yet clear. In view of the complex signal network by cascade reaction and crosstalk of multi-signaling pathways within an organism, and the coordinated regulation of multi-components and multi-targets of traditional Chinese medicine, there may be different components regulating the same signal network or situations where the amount of certain chemical components within a range is not sufficient to cause a change in the signal network. Therefore, the quality control of traditional Chinese medicine based on the effect benchmark may be a useful supplement to the quality standard of traditional Chinese medicine. This paper proposes a Q-biomarker research strategy based on the effect benchmark in order to provide a methodological reference for the quality control research of traditional Chinese medicine.
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To intuitively understand the phenotypic diversity of intra-population and inter-population of the medicinal Cistanche Herba distributed in Xinjiang province, three species of Cistanche Herba were selected for the first time to be conducted to phenotypic observation and measurement from the morphological perspective, aiming to fill the gap in the morphological research concerning Cistanche Herba, and discuss about the relationship between the phenotypic variation and the host plants together with the geographical conditions, thus better understanding the speciation and evolutionary mechanism of Cistanche Herba and providing some scientific basis for the resource protection and germplasm breeding of Cistanche Herba. Based on sampling survey, a total of 118 well grown medicinal Cistanche samples from 17 Cistanche Herba distribution areas in Xinjiang province were selected, and various indexes were observed and measured. Besides, ANOVA and clustering analysis were conducted with 9 phenotypic quantity characters. The Cistanche Herba was plentiful in phenotypic variation. For detail, significant intra-population differences were observed in eight of the nine character indexes, and the intra-population differences were more obvious than those of inter-population. For each quantity character of the three species, the flower density possessed the maximal variable coefficient (71.1%) while the flower length was the minimum (15.9%). The phenotypic variation was also obvious among different populations. Specifically, the average variable coefficient of flower number was the maximal one (46.5%) and the flower length was the minimum one (10.0%). For different populations, the average variable coefficient of the D4 population was the maximal one (41.1%) and the S3 population was the minimum one (13.5%). According to the clustering analysis, all the samples of Cistanche Herba could be divided into three groups when the Euclidean distance was set at 15. The populations of S1, S3, D1, S2, D4, D6, D7 and D5 which distributed in the north of Xinjiang were clustered into one group, and the populations of D8, S4, D9, D2 and D3 that grown in east and central of Xinjiang were clustered into another group. The populations of C. deserticola and C. salsa could not be completely separated, but both of them were obviously differentiated from the T1, T3 and T2 populations of C. tubulosa. Besides, the C. deserticola and C. salsa displayed a patch distribution among different populations, and similar phenotypic characters were shared for each population. The research results of phenotype were consistent with that of molecular biology study of Cistanche Herba. The different phenotypic characters in different distribution areas were deduced to be arose from geographical isolation caused by mountains, which led to the specific genetic structure for each population of Cistanche Herba during the long-term adaptation and evolution. In conclusion, the current study showed the adaptation potency of Cistanche Herba exposed to different habitats.
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Chine , Cistanche , Classification , Analyse de regroupements , Écosystème , Fleurs , Génétique des populations , Phénotype , Plantes médicinales , ClassificationRésumé
To provide molecular evidence for medical material identification, we analyzed the nucleotide sequence of ITS2, psbA-trnH gene in Morus genus plants and commercial products which were obtained from different places in Xinjiang. The sequence of ITS2 and psbA-trnH in fifty-one samples were amplified and sequenced, MEGA 6.0 was used to analyze the intra- and interspecific K-2P distances, neighbor-joining (NJ) tree was used to constructing clustering tree. ITS2 sequence analyzed results showed that there is no intra-specific variation among Morus alba, M. alba var. tatarica and M. nigra, but 13 variations sites were exist between M. alba and M. nigra and their inter-specific K-2P distances was 0.04, which indicated that there had significant variation in them. We didn't find informative variation sites between Morus genus plants and commercial products, and we also found that M. nigra can be distinguished from other two species by NJ Tree. PsbA-trnH analysis results showed there was only one variation site between M. alba and M. nigra, but insertion or deletion variation were remarkable evidence among M. alba, M. alba var. tatarica and M. Nigra. Inter-specific variation was accordance with intra-specific variation of commercial products. So ITS2 and psbA-trnH gene were important marker for M. alba, M. alba var. tatarica and M. nigra identification. This study provided important evidence for Uygur medicine identification and market supervision.
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This study was aimed to investigate the qualitative and quantitative distributions of Lycium ruthenicum resources in the middle and lower reaches of Heihe River, for providing scientific evidence for the protective utilization of the resources in the corresponding geographic region. The outdoor sample plot and quadrat survey, literature search, sample collection, in-house identification and classification were performed by route surveying and visiting to the local natives and/or herb farmers based on the current distribution data of the L. ruthenicum resources in the middle and lower reaches of Heihe River. The distributive pattern of the resources was analyzed using ArcGIS program. The data regarding the category/distributed area and the genetic resources of the L. ruthenicum were collected. The data collected in this study may provide the scientific evidence for the protective utilization of the L. ruthenicum resources in the corresponding geographic region, allowing for the avoidance of the ecological environment from being damaged by improper utilization.
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Apocynum venetum belongs to apocynaceae and is a perennial medicinal plant, its stem is an important textile raw materials. The projection of potential geographic distribution of A. venetum has an important significance for the protection and sustainable utilization of the plant. This study was conducted to determine the potential geographic distribution of A. venetum and to project how climate change would affect its geographic distribution. The projection geographic distribution of A. venetum under current bioclimatic conditions in northern China was simulated using MaxEnt software based on species presence data at 44 locations and 19 bioclimatic parameters. The future distributions of A. venetum were also projected in 2050 and 2070 under the climate change scenarios of RCP2.6 and RCP8.5 described in 5th Assessment Report of the Intergovernmental Panel on Climate Change (IPCC). The result showed that min air temperature of the coldest month, annual mean air temperature, precipitation of the coldest quarter and mean air temperature of the wettest quarter dominated the geographic distribution of A. venetum. Under current climate, the suitable habitats of A. venetum is 11.94% in China, the suitable habitats are mainly located in the middle of Xinjiang, in the northern part of Gansu, in the southern part of Neimeng, in the northern part of Ningxia, in the middle and northern part of Shaanxi, in the southern part of Shanxi, in the middle and northern part of Henan, in the middle and southern part of Hebei, Shandong, Tianjin, in the southern part of Liaoning and part of Beijing. From 2050 to 2070, the model outputs indicated that the suitable habitats of A. venetum would decrease under the climate change scenarios of RCP2.6 and RCP8.5.
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BACKGROUND:Studies have shown that tumor stem ceils are the basis and the main cause of tumor recurrence and chemotherapy failure.Therefore,the research on the drug resistance of tumor stem cells has become a hotspot in the field of stem cell research.OBJECTIVE:To analyze the biological characteristics of side population (SP) cells in the U266 cell lines in relation to drug resistance.METHODS:Flow cytometry was used to monitor the percentage of SP cells in the U266 cell lines.Fluorescence-activated cell sorter (FACS) analysis was used to isolate SP and non-SP (NSP) cells from the U266 cell lines.Further analyses of the cell cycle,multidrug resistant protein,methyl cellulose cloning assay of SP cells and NSP cells were performed.Real-time quantitative PCR was used to determine the expression of ABCG2 and MDR1 genes.RESULTS AND CONCLUSION:There were (2.46±0.35)% SP cells in U266 cells,which were most in the G0 phase.The ratio of G0/G1 in SP cells was (81.50±5.42)%,which was significantly higher than that in NSP cells [(39.85±3.21)%;P <0.05].The positive expression rates of P-gp and ABCG2 in SP cells were signficantly higher than those in NSP cells (P <0.05).The cloning efficiency of SP cells was significantly higher than that of NSP cells (P < 0.05).The mRNA expression of ABCG2 and MDR1 was also significantly higher in SP cells than in NSP cells (P < 0.05).To conclude,a small subpopulation of the isolates of U266 cell lines belong to tumor stem cell-like SP cell subset,most of which are at the G0 phase.ABCG2 and MDR1 genes are highly expressed in SP cells,which particularly plays a important role in the multidrug resistance of multiple myeloma stem cell lines.
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To study the ecological distribution and diversity of endophytic fungi associated with Ferula of medicinal plants in Xinjiang. The endophytic fungi were isolated from roots, stems and leaves of Ferula by microbiology research methods and technology. The endophytic fungi were identified using ITS rDNA sequence analysis and morphology analysis. The composition, diversity and preference of endophytic fungal community were analyzed with Shannon-Wiener biodiversity index (H') and Sorensen coefficient (Cs). A total of 337 strains endophytic fungi were isolated and classified into 38 genera, Alternaria, Aureobasidium and Fusarium were the dominant genera. Among the 337 isolates, the endophytic fungi of F. sinkiangensis were the most, The Shannon-Wiener biodiversity index (H') associated with roots of F. fukanensis was the highest, reached 1.85. The highest Sorensen coefficient ( Cs) was between leaf of F. sinkiangensis and leaf of F. ovina, reached 0.75. From the result, endophytic fungi were widely distributed in six Ferula, there are some notable differences between distribution and composition of the endophytic fungi isolated from different issues and different species of Ferula, show a certain degree of species and tissue preference. The results obtained in this study will provide realistic basis and theoretical basis for further study the secondary metabolites of endophytic fungi associated with Ferula, and the relationship between endophytic fungi and their host plants.
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Biodiversité , Écologie , Endophytes , Ferula , Microbiologie , Champignons , Classification , MétabolismeRésumé
In order to study the spectral reflectance differences of Glycyrrhizae Radix under different growth conditions and lay the foundation for quantitative monitoring of Glycyrrhizae Radix remote sensing images, spectra of Glycyrrhiza species under different growth period and different varieties and different regions were measured by a portable spectrometer. The results showed that the reflectivity of annual G. uralensis was obviously higher than that of the two years plant in the visible light band own to the contents of crown layer chlorophyll. The reflectivity of two years G. pallidiflora was higher than that of G. uralensis in the near infrared band own to the leaf area index and the content of leaf water. The red edge spectrum of annual plant fluctuated largely than that of two years plant due to vegetation coverage and leaf area index. G. pallidiflora grew well than G. uralensis. Under different regions of the Glycyrrhiza species, spectral data analysis showed that within a certain range, the average annual precipitation and average annual evaporation were the major factors to affect the differences of Glycyrrhiza species spectral data under different regions owe to the leaf water content, the higher leaf water content, the lower spectral reflectance. The principal component analysis and continuum-removed method of the spectral data under different regions found that, within a certain range, the average annual precipitation and average annual evaporation were the major factors caused by the differences of Glycyrrhiza species spectral data under the different regions, Glycyrrhiza species spectral similarity related to the spatial distance.
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Géographie , Glycyrrhiza , Chimie , Analyse en composantes principales , Analyse spectraleRésumé
To investigate the resources of medicinal plant, such as wild Apocynum, supervised classification based on Principal Component Analysis (PCA) and texture feature were used to monitor wild medicinal plants from image captured by ZY-3 and World-view-2 and compare which satellite Image are more appropriate to monitor the wild medicinal plants. The research results shows that: for more complex growth conditions wild medicinal plants Apocynum, high-resolution images Worldview-2 is more suitable for its remote identification, the low-resolution satellite ZY-3 can only recognizes the wild medicinal plants which distributed intensively. If the study target distribution is more intensive and larger scale, and cultivated type medicinal plants, the use of satellite ZY-3 in low resolution remote sensing data to identify the target can be a good choice, it is not necessary to buy high-resolution data, in order to avoid waste of expenditure, for the scattered distribution, the high-resolution satellite imagery data may be indispensable to identify targets.
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Apocynum , Chimie , Chine , Conservation des ressources naturelles , Systèmes d'information géographique , Dispersion des plantes , Plantes médicinales , Chimie , Technologie de télédétection , MéthodesRésumé
Morphology and molecular identification technology were used to identify 3 original plants of Fructus Elaeagni which was commonly used in Uygur medicine. Leaves, flowers and fruits from different areas were selected randomly for morphology research. ITS2 sequence as DNA barcode was used to identify 17 samples of Fructus Elaeagni. The genetic distances were computed by kimura 2-parameter (K2P) model, and the Neighbor-Joining (NJ) and Maximum Likelihood phylogenetic trees were constructed using MEGA5.0. The results showed that Elaeagnus angustifolia, E. oxycarpa and E. angustifolia var. orientalis cannot be distinguished by morphological characteristics of leaves, flowers and fruits. The sequence length of ITS2 ranged from 220 to 223 bp, the average GC content was 61.9%. The haplotype numbers of E. angustifolia, E. oxycarpa and E. angustifolia var. orientals were 4, 3, 3, respectively. The results from the NJ tree and ML tree showed that the 3 original species of Fructus Elaeagni cannot be distinguished obviously. Therefore, 3 species maybe have the same origin, and can be used as the original plant of Uygur medicineal material Fructus Elaeagni. However, further evidence of chemical components and pharmacological effect were needed.
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Codage à barres de l'ADN pour la taxonomie , Méthodes , ADN des plantes , Génétique , Espaceur de l'ADN ribosomique , Génétique , Contamination de médicament , Médicaments issus de plantes chinoises , Chimie , Classification , Elaeagnaceae , Classification , Génétique , Fruit , Classification , Génétique , Données de séquences moléculaires , Phylogenèse , Contrôle de qualitéRésumé
To improve accuracy of estimation in planted safflower acreage,we selected agricultural area in Yumin County, Xinjiang as the study area. There safflower was concentrated planted. Supervised classification based on Principal Component Analysis (PCA) and texture feature were used to obtain the safflower acreage from image captured by ZY-3. The classification result was compared with only spectral feature and spectral feature with texture feature. The research result shows that this method can effectively solve the problem of low accuracy and fracture classification result in single data source classification. The overall accuracy is 87.519 1%, which increases by 7.117 2% compared with single data source classification. Therefore, the classification method based on PCA and texture features can be adapted to RS image classification and estimate the acreage of safflower. This study provides a feasible solution for estimation of planted safflower acreage by image captured by ZY-3 satellite.
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Algorithmes , Carthamus tinctorius , Chimie , Traitement d'image par ordinateur , Reconnaissance automatique des formes , Analyse en composantes principales , Méthodes , Technologie de télédétection , MéthodesRésumé
<p><b>BACKGROUND</b>Pancreatic β cells are susceptible to fatty acid-induced apoptosis. The 17β-estradiol (E2) protects pancreatic β cells from apoptosis, mediated by the estrogen receptor-α (ERα). The mRNA level and promoter activity of leukemia-related protein (LRP) 16 were significantly increased by E2 in ER-α and LRP16 was a co-activator of ER-α. The aim of the study was to assess the effects of LRP16 on fatty acid-induced apoptosis in MIN6 cells.</p><p><b>METHODS</b>Cells with over-expressing LRP16 were obtained by lipidosome transfection. Insulin content and glucose-stimulated insulin secretion (GSIS) were examined by radioimmunoassay. Western blotting was applied to detect protein expression. Apoptosis was detected by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) and flow cytometry. The forkhead boxO1 (FoxO1) subcellular localization was determined by immunocytochemical analysis.</p><p><b>RESULTS</b>MIN6-LRP16 cells with overexpression of LRP16 were successfully established, and protein expression of LRP16 was 2.29-fold of that of control cells (MIN6-3.1, P < 0.05). Insulin content and GSIS in MIN6-LRP16 were substantially increased compared with those in control cells. When cells were stimulated with glucose, increased phosphorylation of extracellular signal-regulated kinase (ERK) 1/2 and serine-threonine kinase (Akt) were observed in MIN6-LRP16. When cells were under palmitate pressure, the TUNEL-positive rate in MIN6-LRP16 was (17.0 ± 0.5)%, while it in MIN6-3.1 was (22.0 ± 0.4)%. In palmitate-treated cells, attenuated Akt phosphorylation was observed, but the attenuation in Akt activity was partially restored in MIN6-LRP16 cells. Meanwhile, nuclear localization of FoxO1 in MIN6-LRP16 was apparently reduced compared with that in control cells.</p><p><b>CONCLUSIONS</b>LRP16 regulated insulin content and GSIS in MIN6 cells by ERK1/2 and Akt activated way. Meanwhile, LRP16 overexpression protected MIN6 cells from fatty acid-induced apoptosis by partially restoring Akt phosphorylation and inhibiting FoxO1 nuclear redistribution. Therefore, LRP16 played important roles not only in insulin content and GSIS but also in the antilipotoxic effect mediated by Akt/FoxO1 signaling.</p>
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Animaux , Souris , Apoptose , Technique de Western , Lignée cellulaire tumorale , Acides gras , Pharmacologie , Protéine O1 à motif en tête de fourche , Facteurs de transcription Forkhead , Génétique , Métabolisme , Protéines tumorales , Génétique , Métabolisme , Phosphorylation , Protéines proto-oncogènes c-akt , Génétique , Métabolisme , RT-PCR , Transduction du signal , GénétiqueRésumé
The aim of this study was to investigate the effect of different concentrations of arsenic trioxide (As(2)O(3)) on apoptosis of lymphoblastoid Raji cell line and its possible mechanisms, as well as its relation to the expression of mcl-1 gene. The Raji cells were treated with different concentrations of As(2)O(3), the classical DNA ladder of cell apoptosis was detected by agar gel electrophoresis, the changes of mitochondrial membrane potential in Raji cells were assayed by confocal laser scanning microscopy, the changes of mcl-1 gene expression after exposure of cells to As(2)O(3) was detected by real-time quantitative fluorescence PCR. The results indicated that 1 micro mol/L As(2)O(3) did not lead to significant apoptosis of Raji cells, 2 - 8 micromol/L As(2)O(3) induced Raji cell apoptosis. Along with increase of drug concentration, the mitochondrial respiratory function and membrane potential of Raji cells obviously decreased. At same time, the expression level of mcl-1 gene were significantly down-regulated. It is concluded that As(2)O(3) can markedly decrease mitochondrial respiratory function and membrane potential of Raji cells, and down-regulate expression of mcl-1 gene, which may be the mechanisms resulting in cell apoptosis.
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Humains , Antinéoplasiques , Pharmacologie , Apoptose , Composés de l'arsenic , Pharmacologie , Lignée cellulaire tumorale , Expression des gènes , Potentiel de membrane mitochondriale , Mitochondries , Métabolisme , Protéine Mcl-1 , Oxydes , Pharmacologie , Protéines proto-oncogènes c-bcl-2 , GénétiqueRésumé
This study was aimed to investigate the JAK2V617F mutation in myeloproliferative disorders (MPD) and to evaluate the significance of JAK2V617F in diagnosis and therapy of MPD. The bcr/abl fusion gene in 70 MPD patients was detected by reverse transcription polymerase chain reaction (PT-PCR). The JAK2V617F mutation was detected by allele-specific polymerase chain reaction (AS-PCR) and the results were confirmed by sequence analysis. The results indicated that the bcr/abl fusion gene could be detected in 38 patients with chronic myeloid leukemia (CML), but not in the 32 none-CML patients. The JAK2V617F mutation was detected in 12 out of 16 (75%) patients with polycythemia vera (PV), 3 out of 10 (30%) patients with essential thrombocythemia (ET), 3 out of 6 (50%) patients with idiopathic myelofibrosis (IMF), but not in any of the CML patients. The JAK2V617F mutation frequencies between CML and bcr/abl negative MPD patients were statistically significant (p < 0.05). It is concluded that the JAK2V617F may be a characteristic molecular event in PV, ET and IMF patients which may serve as an important molecular marker for the diagnosis and classification of the three diseases.
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Adulte , Sujet âgé , Sujet âgé de 80 ans ou plus , Femelle , Humains , Mâle , Adulte d'âge moyen , Kinase Janus-2 , Génétique , Syndromes myéloprolifératifs , Génétique , Mutation ponctuelle , Réaction de polymérisation en chaîne , MéthodesRésumé
This study was purposed to set up real-time quantitative RT-PCR technique and to measure leukemia fusion gene transcripts in patients with chronic myeloid leukemia (CML), acute lymphoblastic leukemia (ALL) and acute promyelocytic leukemia (APL). All plasmids containing the target gene sequences were constructed to establish the standard curves. A TaqMan based real-time quantitative RT-PCR was performed to measure aberrant fusion gene transcripts in 130 samples of peripheral blood (PB) or bone marrow (BM) from 49 patients with leukemia. The results showd that the BCR-ABL(P210) transcripts were detected in 28 (82.4%) out of 34 CML patients (the ratios of BCR-ABL(P210)/ABL varied from 0.01 to 3.19) and also in 2 (33.3%) out of 6 ALL patients. The BCR-ABL(P190) transcripts were detected in 2 (33.3%) out of 6 ALL patients. The BCR-ABL(P210) and BCR-ABL(P190) transcripts were both detected in 1 (2.9%) CML patients. The PML/RARalpha transcripts were detected in 7 (77.8%) out of 9 APL patients (the ratio of PML-RARa/ABL varied from 0.0014 to 3.199). The relative frequency of both bcr1 and bcr3 was 42.9%, while that of bcr2 was 14.3%. The transcript level of aberrant fusion gene varied from the clinical situation of patient. It is concluded that real-time quantitative PCR is a reliable, innovative and promising technology with high sensitivity and specialty. It has potential clinical value for defining diagnosis, typing tumor, selecting treatment, measuring the tumor load, monitoring fusion gene expression level and evaluating therapeutic strategies, which is worthy to be popularized.