OBJECTIVE:To explore the clinical and laboratory characteristics of 5 patients with myeloid leukemia and t(12;22)(p13;q12).METHODS:Bone marrow cells were cultured for 24 h and analyzed by standard R-banding. Rearrangement of the MN1 gene was detected by fluorescence in situ hybridization (FISH) using dual color break-apart MN1 probes. MN1-ETV6 and ETV6-MN1 fusion genes were detected by reverse transcription polymerase chain reaction (RT-PCR). And the products were subjected to direct sequencing.RESULTS:Among the 5 patients, 2 had AML-M0, 2 had AML-M4, and 1 had CMM0L at the initial diagnosis. t(12;22)(p13;q12) was the primary abnormality among all patients. Rearrangements of MN1 gene were detected by FISH in all patients. MN1-ETV6 and ETV6-MN1 fusion genes were detected respectively in 4 and 3 patients.CONCLUSION:t(12;22)(p13;q12) is a rare but recurrent chromosomal abnormality in myeloid leukemia, and is related to poor prognosis. allo-SCT is valuable for patients with t(12;22)(p13;q12).

DOI:10.3760/cma.j.issn.0253-2727.2018.09.014 作者单位:215006 苏州大学附属第一医院、江苏省血液研究 所;卫生部血栓与止血重点实验室 通信作者:潘金兰,Email:jinlanpan@126.com Phlike acute lymphocytic leukemia with EPOR rearrangement: 2 cases report and literatures review Yang Qian, Shao Haigang, Liu Hengfang, Bai Shuxiao, Zhang Jun, Wang Yong, Shen Juan, Wu Chunxiao, Qiu Huiying, Chen Suning, Pan Jinlan Corresponding author: Pan Jinlan, Jiangsu Institute of Hematology, the First Affiliated Hospital of Soochow University, Thrombosis and Hemostasis Key Laboratory of the Ministry of Health, Soochow University, Suzhou 215006, China.Email: jinlanpan@126.com

OBJECTIVE:To report clinical and laboratory features of 4 cases of myeloid neoplasm with t (5;12) (q33;p13).METHODS:Cytogenetic examination of bone marrow cells obtained from patients was performed by 24 h culture method. R banding technical was used for karyotype analysis. PDGFRβ gene rearrangement was detected by FISH using dual color break apart PDGFRβ probe. ETV6-PDGFRβ fusion genes were detected by multiple-reverse transcription polymerase chain reaction (RT-PCR). Direct sequencing analysis was performed on the PCR products in case 1. Immunophenotype analysis was carried out by flow cytometry. Four cases were treated with imatinib (IM) and followed up.RESULTS:The diagnoses included 3 MPN and 1 AML-M2. The t (5;12) (q33;p13) was a primary abnormality in 3 cases of MPN and a secondary abnormality in 1 case of AML-M2. PDGFRβ gene rearrangement and ETV6-PDGFRβ fusion genes were detected by FISH and multiple-RT-PCR in 4 cases, respectively. The immunophenotypical analysis of leukemia cells showed positive for CD13, CD33 and CD34. Two cases obtained MMR after the treatment of IM, one case complete hematologic and complete cytogenetic response. ETV6-PDGFRβ was negative detected by multiple-RT-PCR after the treatment of IM, but relapsed and died soon in case 4.CONCLUSIONS:The t (5;12) myeloid neoplasm was a subtype with unique features. The t (5;12) maybe a primary chromosome abnormality in MPN and a secondary in AML. MPN with t (5;12) could benefit from IM, but not for AML. Dual-FISH was a reliable tool for detecting PDGFRβ rearrangement.

This study reports 10 patients with hematological malignances with t(20;21)(q11;q11) resulting from del(20q) (for example, der(20)del(20)(q11q13)t(20;21)(q11;q11) and der(21)t(20;21)(q11;q11)) and described their clinical features and the possible prognostic significance of this abnormality. The t(20;21)(q11;q11) was a rare but recurrent abnormality secondary to del(20q) besides i(20q-). The frequency of der(20)del(20)(q11q13)t(20;21)(q11;q11) among our patients with del(20q) was 2.4%. It was considered that the 20q deletion preceded translocation with chromosome 21. This abnormality is often cryptic, occurs predominantly in older men and is observed most often in myelodysplastic syndromes. Patients with this abnormality have an unfavorable prognosis, similar to patients with i(20q-). The molecular consequences of der(20)del(20)(q11q13)t(20;21)(q11;q11) may be different from patients with i(20q-). To the best of our knowledge this is the largest dataset published to date.

Deletion of the long arm of chromosome 20 is a common abnormality underlying hematological malignancy. We analyzed 21 patients with hematologic diseases confirmed to carry the del(20q) by conventional cytogenetics and fluorescence in situ hybridization using microarray comparative genomic hybridization (aCGH). Seventeen patients were positive for del(20q), but this deletion was not detected in four patients. All deletions detected were interstitial of which continuous deletions were seen in 12 patients and discrete deletions in five. Three commonly deleted regions (CDRs) and two commonly retained regions (CRRs) were defined: CDR1 spanning 3.05Mb (34560497-37608229) within 20q11.23, CDR2 spanning 1.76Mb (37851501-39615698) within 20q12, CDR3 spanning 116Kb (48120412-48236791) within 20q13.13, CRR1 spanning 1.1Mb (29374726-30428250) within 20q11.21, and CRR2 spanning 2.5Mb (60484668-62963548) within 20q13.33. Duplications of retained regions (20q11.21) were found in five cases with similar erythroid hyperplasia (2 M6, 3 MDS). Moreover, duplication of 20p13-p11.21 was also found in two cases with M6. Using the CDRs and CRRs, we identified the candidate genes we searched for using the UCSC Genome Browser. Our data suggest that aCGH analysis is useful for more precisely defining breakpoints on 20q. Further work is required to identify candidate pathogenic genes within these CDRs and CRRs.

Owing to the important utilities in many areas, the preparation of chiral sulfur-containing compounds has attracted special attention of chemists. As an alternative of the most popular asymmetric sulfur-Michael addition, asymmetric alpha-sulfenylation has been developed well in recent years. This short review would summarize the advance on asymmetric alpha-sulfenylation. Two asymmetric protocols, including transition-metal catalysis and organocatalysis, are discussed respectively.

The clinical and hematological characteristics and the prognostic significance of del(20q) were investigated in a consecutive series of 213 myeloid malignancies. In the analyses, the cases were divided into three subgroups according to diagnosis or four subgroups according to cytogenetic data. Patients in the myeloproliferative neoplasms subgroup had high WBCs and platelet counts at initial diagnosis. The del(20q) occurred predominantly in older men. Sole del(20q) was observed most often in myelodysplastic syndromes, while del(20q) as a part of complex karyotypes was observed predominantly in acute myeloid leukemia. The most frequent additional abnormalities accompanying del(20q) were -5/del(5q), -7/del(7q) and +8; t(20;21)(q11;q11) and double del(20q) were two rare but recurrent abnormalities secondary to del(20q). In all types of diseases, patients with a sole del(20q) had a favorable prognosis. The presence of any additional abnormality with del(20q) had an unfavorable outcome. Patients with i(20q) had an unfavorable prognosis. Patients with the minor del(20q) clone had a better median survival than those with the major del(20q) clone.

The proton is the smallest functional group in organic synthesis and it is extremely challenging to control it in terms of enantioselectivity. In recent years, the use of organocatalysts has witnessed rapid developments and has become the state-of-art in enantioselective protonation of transient enolate. The key prochiral enolate can be generated via various reaction types. Herein we review recent reports of organocatalytic enantioselective protonation of transient enolate, classifying them according to the type of enolate precursors.

It is our great pleasure as Guest Editors of the journal ‘Mini-Reviews in Organic Chemistry’ to present you with a ‘mini-hot topic issue’ on nonmetal asymmetric catalysis. Asymmetric catalysis entails the catalytic, selective, and reproducible generation of a given enantiomer of a chiral product from achiral reactants. Leading the quest of asymmetric catalysis is the need of the pharmaceutical, flavors and fragrances, and agrochemical industries for enantiopure molecules because the different enantiomers or diastereomers of a molecule often have different biological activity. Nearly 85% of new drugs in the market are chiral. Organocatalysis can be used as environmentally-friendly alternatives to transition-metal catalysts as no toxic metals are required. The principle interactions of organocatalyst are non-covalent, such as hydrophobic, hydrogen bonding, van der Waals and electrostatic as in enzymes. These catalysts are often inexpensive to prepare and reaction can be performed under aerobic environments and in wet solvents. The beneficial impact of organocatalytic reactions on a large scale production of chiral intermediates has been demonstrated. Therefore, this special issue aims to review the advances in nonmetal asymmetric catalysis during the last decade, specified in organocatalysis. The first review related to the above mentioned topic is written by Drs. Leow, Shen and their colleagues, who explore organocatalytic enantioselective protonation of enol derivatives. As they mentioned, the proton is the smallest functional group in organic synthesis and it is extremely challenging to control it in terms of enantioselectivity. Traditional methods are using pre-formed enolates obtained from the direct deprotonation of carbonyl compounds by strong bases. The newer strategies involve the usage of silyl enol ethers, tautomerization of enols or in situ generation of transient enolates. They can be generated through either additions to ketenes, conjugate additions to Michael acceptors, rearrangements, or isomerization of double/triple bonds. In the following article, as an alternative of the most popular asymmetric sulfur-Michael addition, Dr. Zhiyong Jiang and his colleagues summarized the advance on asymmetric α-sulfenylation. Two asymmetric protocols were discussed respectively, including transition-metal catalysis and organocatalysis. It was noted that the unprecedented substrate scopes still exist which represents developing rooms, although asymmetric α-sulfenylation has obtained remarkable progress in recent years; and novel and easily prepared sulfenylated reagents are still highly desirable. In the last article in this issue, Dr. Hansen and his colleagues gave an overview over the major advances in asymmetric enamine catalysis since, and including, 2008 with focus on strategic transformations and catalyst development. As they noted, enamine-catalysis has become a powerful enabling technology for enantioselective chemical synthesis, which could be attributed to a rapid evolution of novel catalyst architectures and strategies, combined with the design and discovery of strategically important, asymmetric transformations. This exciting field will continue to expand, fuelled by its unique ability to generate novel and stereodefined structural complexity in the synthesis of organic molecules. We would like to sincerely thank all the reviewers for their valuable contributions that ensured the quality of articles to be published in this special issue. It was a great opportunity for us to cooperate with researchers from all over the world including United States, Norway, Taiwan and Mainland China. We hope that readers will enjoy this issue, obtain useful information, and be inspired with new ideas for future research on nonmetal asymmetric catalysis.

The first organocatalytic asymmetric Michael addition of 5H-oxazol-4-ones to nitroolefins has been developed. In the presence of easily prepared L-tert-leucine-derived tertiary amine/thiourea catalyst, the Michael addition of 5H-oxazol-4-ones to nitroolefins proceeded in an excellent diastereo- and enantioselective manner (up to 99% ee and >19:1 dr). The Michael adducts obtained are valuable precursors for the synthesis of chiral α-alkyl-α-hydroxy carboxylic acid derivatives, which represent a series of versatile building blocks in many biologically active compounds.

OBJECTIVE:To analyze clinical and cytogenetic features of hematological disorders associated with 20q- and t (20;21) (q11;q11) abnormalities.METHODS:Following short-term culture of bone marrow cells, karyotypic analysis was carried out with R-banding. 20q- and t(20;21) (q11;q11) was detected by fluorescence in situ hybridization (FISH) using dual-color 20q11/12 probe, ST 20qter /ST 21qter probes, SE20(D20Z1)/SE 13/21 probes, and WC20/WC21 probes.RESULTS:Six (2.3%) of the 257 patients with 20q- detected by conventional karyotypic analysis were found to have t(20;21) (q11;q11) abnormality. Five cases had myelodysplastic syndrome, 1 had acute lymphoblastic leukemia. Above results were all confirmed by FISH.CONCLUSION:i (20q-), t(20;21) (q11;q11) seems to be a rare but recurrent chromosomal abnormality which is specifically associated with myeloid disease, late occurrence and poor prognosis. The translocation between chromosome 20q11 and 21q11 may form a novel fusion gene which has an important role in the pathogenesis of the disease.

This study was aimed to summarize and analyze the morphology, immunophenotype, cytogenetics, molecular biology (MICM), tyrosine kinase (TK) gene mutations and clinical features of acute myeloid leukemia (AML) with complex variant of t(8;21). A retrospective study was performed for 20 AML patients with complex variant of t(8;21) in our hospital from January 1994 to April 2012, including analysis of clinical feature, immunophenotype, chromosome karyotype, treatment regimen, as well as the overall survival (OS) and relapse-free survival (RFS). Mutations of C-KIT, FLT3-ITD, FLT3-TKD and JAK2V617F were detected by genomic DNA PCR and the sequencing was per-formed in 13 AML patients with complex variant of t(8;21). The results showed that (1) the incidence of 20 AML patients with complex variant of t(8; 21) was 2.4% of total t(8; 21) AML patients. In 20 AML patients with complex variant of t(8;21), 1 case was M1, 17 cases were M2, 2 cases were M4; 10 cases were myeloid phenotype and the other 3 were myeloid plus lymphoid phenotype. There were 16 kinds of cytogenetics additional involvement of chromosomal breakpoints: lp22, 1p32, 2q35, 2q14, 3p25, 5q13, 6p22, 7q21, llq11, 1lq13, 12q14, 12q24, 12p12, 14q32, 15p13, 20q12. (2) C-KIT aberrations were detected in 30.8% cases, all mutated in exon 17 (mutkit 17), only 1 case had JAK2V617F mutation. The result of FLT3 mutation screenings in AML patients with complex variant of t(8; 21) was negative. Of 5 patients with gene mutations, 1 patient (20%) achieved complete remission (CR), the median RFS and median OS time were 6.5 months and 8.9 months respectively. Of the 8 patients without gene mutations, 6 patietns (75%) achieved CR; the median RFS and median OS time were 26.6 months and 27.7 months respectively. It is concluded that the AML patients with complex variant of t(8;21) shows typical features of t(8;21) AML, but the existence of the tyrosine kinase-related gene mutation has important implications on remission rate and long-term survival of patients treated by induction chemotherapy.

OBJECTIVE:To report the clinical and laboratory characterization of a case of multiple myeloma with low hypodiploid complex karyotyptic abnormalities.METHODS:Cytogenetic examination of bone marrow performed by 24 h culture method. R-banding technique was used to analyze the karyotype. Interphase fluorescence in situ hybridization (FISH) was performed using chromosome probes such as 13q14, p53, Rb1, 1q21 and IgH/CCND1. The DNA content was detected by flow cytometry.RESULTS:Chromosome analysis revealed complex chromosomal rearrangement. Five cells had a low hypodiploid karyotype with 35 chromosomes. Three cells had the duplication of the low hypodiploid karyotype. Four cells had a normal karyotype. Monosomy 1, 13, 14, 17 and a mark chromosome 1 derived from chromosome 11 resulting in the amplication of CCND1 gene were confirmed by interphase FISH. Flow cytometric analysis displayed a low hypodiploid peak with the DNA index of 0.8426.CONCLUSION:These results indicated that the low hypodiploidy is a rare abnormality in multiple myeloma. Interphase FISH is a reliable method for detecting molecular abnormalities in multiple myeloma.

Objective: To investigate the clinical and laboratory characteristics of one patient with AML (acute myeloid leukemia) carrying t(7;21)(p21;q22) — a new variant form of t(8;21) (p21;q22).Methods: Cytogenetic examination was performed after culture of cells from bone marrow for 24 h. R-banding technique was used for karyotype analysis. AML1/ETO fusion gene was detected by dual-color fluorescence in situ hybridization (FISH) using AML1/ETO dual-color probes. Quantitative analysis of AML1/ETO chimeric transcripts was performed by using a real-time fluorescence-based quantitative PCR.Results: Chromosome analysis revealed a karyotype of 46, XX, t(7;21)(p21;q22),and AML1/ETO fusion gene was found in 86% of the bone marrow cells. The AML1/ETO fusion transcript copy number was 51 440/10 000 Abl copies.Conclusion: The AML patient carrying t(7;21)(p21;q22) which is a new variant form of t(8;21)(q22;q22) may have a better prognosis. DOI:10.3781/j.issn.1000-7431.2012.10.013