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Objective:The research group found in the early stage that the 75%alcohol extract of the Cinnamomi Ramulus had a significant physiological activity in inhibiting necroptosis by screening out the self-built sample library of 100 kinds of traditional Chinese medicines in Jiangxi. To identify the active components and find the target compounds,the 75%alcohol extracts of Cinnamomi Ramulus were isolated and studied systemically in chemistry. Method:The 20 kg dry Cinnamomi Ramulus was crushed into coarse powder,and extracted with 75%alcohol for four times, one time every 7 d. Then total extracts were obtained after solvent was recycled under decompression. The extract was separated by D101 macroporous resin column chromatography and eluted by water,30%ethanol,50%ethanol,70%ethanol,90%ethanol,so as to get the corresponding fraction finally. The compounds in the 30%ethanol and 50%ethanol fraction were isolated and purified by chromatography on silica gel,Sephadex LH-20 column and high pressure preparative chromatography,and their structures were determined according to physicochemical properties and spectral analysis. Result and Conclusion:Thirteen compounds were isolated and identified as (+)-syringaresinol (1),(+)-lyoniresinol (2),spicatolignan B (3),(-)-secoisolariciresinol (4),ovafolinin B (5),protocatechualdehyde (6),protocatechuic acid (7),syringaldehyde (8),vanillic acid (9),ethyl protocatechuate (10),syringic acid (11),ethyl gallate (12),2-(3',4'-dihydroxyphenyl)-1,3-pepper ring-5-aldehyde (13). Compounds 1-5,10-13 were isolated from this plant for the first time.
Résumé
<p><b>OBJECTIVE</b>To study the clinical and laboratory features of childhood acute leukemia (AL) with MLL gene rearrangements.</p><p><b>METHODS</b>Sixteen of 298 cases of childhood AL with MLL rearrangements were studied by using MLL dual-color FISH, multiplex RT-PCR with 13 pairs of primers in combination with R banding karyotype analysis and cell immunophenotyping by flow cytometry.</p><p><b>RESULTS</b>Sixteen cases of childhood AL with MLL rearrangements accounted for 5.4% of 298 AL patients, and 56.3% of infant ALs. Among 106 cases analyzed by multiplex RT-PCR, MLL gene rearrangements were found in 11 cases, including MLL/AF4 fusion gene in 2, MLL/AF6 fusion gene in 1, MLL/AF6 and MLL/ELL combined with MLL/ AFX or HOX11 in one case each, MLL/AF9 in 2, MLL/AF10 in 1, MLL/ELL in 2. MLL partial tandem duplication in 1 and activated HOX11 in 1. In 27 cases assayed by FISH, 9 cases (36.0%) were demonstrated MLL gene rearrangements. In 16 patients with MLL gene rearrangements, 14 (87.5%) exhibited clonal chromosome abnormalities involved chromosome 11 in 11 cases: being t(4;11) in 2, t(6;11), t(8;11), t(7;8;11), t(9;11) in each trisomy 11 in 2 and 11q--in 3 cases. Among these 16 patients, 11 were B-ALL, and 5 AML-M5, 3 of the latter were CD7+ and CD2+. Of these 16 patients, 8 received chemotherapy and 7 of them achieved complete remission, while the other 8 patients gave up treatment.</p><p><b>CONCLUSION</b>Multiplex RT-PCR combined with FISH provided a more accurate and sensitive method for detection of MLL gene rearrangements. Finding out MLL gene rearrangement is of most importance in guiding therapy and predicting prognosis in childhood AL.</p>
Sujets)
Adolescent , Enfant , Enfant d'âge préscolaire , Femelle , Humains , Nourrisson , Mâle , Réarrangement des gènes , Hybridation fluorescente in situ , Leucémies , Génétique , Protéine de la leucémie myéloïde-lymphoïde , GénétiqueRésumé
<p><b>OBJECTIVE</b>To detect the expression of the fusion genes resulting from chromosome abnormalities in childhood acute lymphoblastic leukemia(ALL) and its conformity to WHO classification.</p><p><b>METHODS</b>Sixty-two children with ALL were investigated. The expression of fusion genes was determined by multiplex reverse transcription-polymerase chain reaction (RT-PCR), karyotyping (R band) and immunophenotyping (by flow cytometry) were also performed.</p><p><b>RESULTS</b>Of the 62 patients, 23(37.1%) were found to carry 13 different fusion genes. The patients with immunophenotype of Pre-B-ALL were found to carry: TEL/AML1(3 cases); E2A/PBX1, E2A/HLF, TLS/ERG, MLL/AF4, MLL/AF9, MLL/AF10, MLL/AFX-MLL/AF6-MLL/ELL, MLL/AF6-MLL/ELL, dupMLL (one case for each); and HOX11 (6 cases). The patients with immunophenotype of Pre-T-ALL were found to carry: TAL1D (4 cases, one is also found to have HOX11 expression); and HOX11 (2 cases). The multiplex RT-PCR in combination with chromosome analysis revealed genetic abnormalities in 69.4%(43/62) of childhood ALL.</p><p><b>CONCLUSION</b>Multiplex RT-PCR combined with chromosome analysis and immunophenotyping can provide reliable and helpful information for the diagnosis, therapy evaluation and prognosis prediction in childhood ALL, which may also serve as a basis on which to implement the criteria of WHO classification.</p>
Sujets)
Adolescent , Enfant , Enfant d'âge préscolaire , Humains , Nourrisson , Aberrations des chromosomes , Sous-unité alpha 2 du facteur CBF , Génétique , Métabolisme , Protéines de liaison à l'ADN , Génétique , Métabolisme , Cytométrie en flux , Protéines à homéodomaine , Génétique , Métabolisme , Immunophénotypage , Caryotypage , Protéine de la leucémie myéloïde-lymphoïde , Génétique , Métabolisme , Protéines de fusion oncogènes , Génétique , Métabolisme , Leucémie-lymphome lymphoblastique à précurseurs B et T , Génétique , Métabolisme , Protéines proto-oncogènes , Génétique , Métabolisme , Protéine FUS de liaison à l'ARN , Génétique , Métabolisme , RT-PCR , Facteurs de transcription , Génétique , MétabolismeRésumé
<p><b>OBJECTIVE</b>To study the value of combination assay of multiplex RT-PCR and karyotypic analysis in the diagnosis and classification of childhood acute lymphoblastic leukemia (ALL).</p><p><b>METHODS</b>Fifty cases of childhood ALL patients were studied by multiplex RT-PCR in combination with R or G banding karyotype analysis.</p><p><b>RESULTS</b>Of the 50 childhood ALL patients, 18 (36.0%) carried 11 types of fusion genes including E2A/PBX1, TEL/AML1, TLS/ERG, MLL/AF4, MLL/AF9, MLL/AF10, MLL/AFX, MLL/AF6, MLL/ELL, TAL1D, and HOX11, revealed by multiplex RT-PCR, and in 48 cases, 24 (57.1%) had chromosome abnormalities. Among the latter, numeral chromosome abnormalities and chromosome deletions accounted for 75.0% (18/24), while translocations 25.0% (6/24). The multiplex RT-PCR in combination with chromosome analysis could detect genetic abnormalities in 70% (35/50) of childhood ALL.</p><p><b>CONCLUSIONS</b>Multiplex RT-PCR combined with chromosome analysis can enhance the detection rate of genetic abnormalities in childhood ALL. It provides reliable evidence for the diagnosis, classification and prognosis.</p>
Sujets)
Adolescent , Enfant , Enfant d'âge préscolaire , Femelle , Humains , Nourrisson , Mâle , Aberrations des chromosomes , Caryotypage , Protéines de fusion oncogènes , Génétique , Leucémie-lymphome lymphoblastique à précurseurs B et T , Classification , Diagnostic , Génétique , Reproductibilité des résultats , RT-PCR , Méthodes , Sensibilité et spécificitéRésumé
<p><b>OBJECTIVE</b>To investigate the interrelations among morphology, immunology, cytogenetics and clinical outcome in childhood acute leukemia with 11q23 abnormalities.</p><p><b>METHODS</b>Eighteen patients with 11q23 abnormalities, from 320 childhood acute leukemia patients, were retrospectively analysed for cell morphology, flow cytometry, immunophenotyping, R-banding karyotype as well as clinical features and prognosis. Twenty cases of childhood AL with normal karyotype during the same period were used as control.</p><p><b>RESULTS</b>The incidence of 11q23 abnormalities in our childhood acute leukemia patients was 5.63% including 14 acute lymphoblastic leukemia (ALL) and 4 acute myeloid leukemia (AML). Of 16 cases immunophenotypically tested, 13 expressed lymphoid antigens and 3 CD(34) and other myeloid antigens. Karyotype analysis disclosed the following abnormalities: t(4; 11)(q21; q23) in 6 cases, t(10; 11)(p13; q23) in 3, t(11; 19)(q23; p13) in one and del(11)(q23) in 6. The complete remission rate for these patients with 11q23 abnormalities was comparable to that of the control (72.2% vs 80.0%, P > 0.05), while the mortality rate in the former was significantly higher than that in the latter (61.1% vs 25.0%, P < 0.05).</p><p><b>CONCLUSIONS</b>11q23 abnormalities were mainly seen in childhood ALL and acute monocytic leukemia with unique prognostic features.</p>