Objective To analyze the mutation status of K‐ras gene in colorectal cancer patients ,further more ,to provide guid‐ance for personalized therapy for colorectal cancer .Methods Nested and COLD‐PCR were used to detect the K‐ras mutations in 560 patients with colorectal cancer .Results In 560 colorectal cancer patients ,the total positive rate of K‐ras gene mutations was 27 .08% ,the mutation rate was 0 in 128 plasma samples and it was 27 .08% in 432 tissue samples .The mutate sites were G12S , G12C ,G12D ,G12A ,G12V ,G13R ,G13C ,G13D ,Q61K ,Q61L ,there were significant differences existed in different samples (P <0 .000 1) ;the mutation rate of 362 male patients was 20 .44% and the types of mutation include G12S ,G12C ,G12D ,G12V ,G13R , G13C ,G13D ,Q61K and Q61L .The mutation frequency was 21 .72% in 198 female patients ,the mutation points were G12S ,G12C , G12D ,G12A ,G12V ,G13R and G13D .There were no significant difference between different sex (P= 0 .722 7) ;the mutation fre‐quency was 20% in 80 youth patients including G12S ,G12C ,G12D ,G12V ,G13D and the mutation rate was 33 .07% in 127 middle age patients ,the points of mutation were G12S ,G12D ,G12A ,G12V ,G13R ,G13C ,G13D ,Q61K ,Q61L ,the mutation frequency was 16 .71% in 353 old age patients ,the types of mutation include G12C ,G12D ,G12V ,G13R ,G13D ,the difference was significant a‐mong different age patients (P= 0 .000 5) .Conclusion The total rate of mutations is 27 .08% in 560 colorectal cancer patients ,and the main points of mutation is G12D ,G12V ,G13D .There are significant differences in different type of samples as well as in differ‐ent ages ,but no statistical significance in different sex patients .

Base Sequence ; Child ; Female ; Heart Septal Defects, Atrial ; complications ; diagnosis ; Humans ; Molecular Sequence Data ; Noonan Syndrome ; complications ; diagnosis ; genetics ; Protein Tyrosine Phosphatase, Non-Receptor Type 11 ; genetics

OBJECTIVETo report clinical and laboratory features of 4 cases of myeloid neoplasm with t (5;12) (q33;p13).

METHODSCytogenetic 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.

RESULTSThe 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.

CONCLUSIONSThe 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.

Chromosome Banding ; Gene Rearrangement ; Hematologic Neoplasms ; genetics ; Humans ; Immunophenotyping ; In Situ Hybridization, Fluorescence ; Karyotyping ; Myeloproliferative Disorders ; genetics ; Polymerase Chain Reaction ; Proto-Oncogene Proteins c-ets ; genetics ; Receptor, Platelet-Derived Growth Factor beta ; genetics ; Remission Induction ; Repressor Proteins ; genetics ; Translocation, Genetic

Objective: To investigate the overexpression frequencies of BRE and EVI1, the correlation between BRE and EVI1 expressions and their possible clinical implications in 11q23/MLL rearrangement acute leukemia. Methods: Cytogenetic examination of bone marrow cells was performed by short-term culture method. R-banding technique was used for karyotype analysis. 47 patients were detected by interphase fluorescence in situ hybridization (FISH) with dual-color break apart MLL probe. The expressions of EVI1 and BRE genes were detected by real time quantitative reverse transcription polymerase chain reaction (RQ-PCR) . The correlation and prognostic significance were statistically tested. Results: 11q23/MLL rearrangements were confirmed by karyotyping and FISH, respectively in 47 patients. According to immunophenotypic analyses of 37 patients, 5 patients showed positive for CD19, CD79a or CD10, 1 for CD7; the others for CD33, CD13, CD14 and CD15, and 16 of them for CD34. Of the 47 patients, 18 patients showed EVI1 overexpression and most of them presented with t (6;11) and M₄/M₅. The EVI1 expression was high in t (6;11) or t (9;11) subgroup comparable with levels observed in normal subgroup (P=0.038, 0.022, respectively) . 15 patients showed high BRE expression, and most of them presented with t (9;11) and M₄/M₅. High BRE expression was found in t (4;11) , t (6;11) , t (9;11) and t (11;19) subgroups, respectively by comparing with normal subgroup. The BRE expression was higher in t (4;11) (P=0.004) or t (9;11) (P=0.012) subgroup than in t (6;11) subgroup. Patients with EVI1 overexpression had a short survival compared with those with low EVI1 expression (P=0.049) and it also did in t (9;11) subgroup (P=0.024) . Patients with t (9;11) and high BRE expression had a long survival compared with those with t (9;11) and low BRE expression (P=0.024) . Conclusion The EVI1 overexpression was significantly frequent in acute leukemia patients with 11q23/MLL rearranged, especially within t (6;11) subgroup and M₄/M₅, which was associated with an inferior outcome. High BRE expression was observed frequently in 11q23/MLL-rearranged acute leukemia especially within t (9;11) subgroup and M₅.

Objective: To study the value of unmethylated cytosine guanine dinucleotide oligodeoxynucleotide (DSP30) and IL-2 in the conventional cytogenetic (CA) detection of the chromosomal aberrations in chronic lymphocytic leukemia (CLL) . Methods: Bone marrow or peripheral blood cells of CLL patients were cultured with DSP30 plus IL-2 for 72 h, following which R-banding analysis was conducted. Fluorescence in situ hybridization (FISH) was performed in 85 patients. CA results were compared with data obtained by FISH. Results: Among 89 CLL patients, the success rate of chromosome analysis was 94.38% (84/89) . Clonal aberrations were detected in 51 patients (51/84, 60.71%) . Of them, 27 (27/51, 52.94%) were complex karyotype. Among 85 CLL patients tested by FISH, chromosomal abnormalities were detected in 74 (74/85, 87.06%) patients, of which 2 (2/74) patients were complex karyotypes, accounting for 2.70%. Of the 85 CLL patients examined by FISH, 50 had abnormal karyotype analysis, 30 had normal karyotype, 5 failed to have chromosome analysis. Among them, 25 cases showed clonal aberrations by FISH assay but normal by CA, and 4 cases were normal by FISH but displayed aberrations in chromosome analysis, and totally 78 (91.76%) cases with abnormality detected by the combination of the two methods. The frequency of 13q- abnormality detected by FISH was significantly higher than that by CA analysis (69.41%vs 16.67%, P<0.001) , while the frequency of 11q-,+12 and 17p- detected by two methods showed no significant difference (P>0.05) . The detection rate of complex abnormalities in conventional karyotype analysis was higher than that in FISH (50.98%vs 2.70%) . In addition, 11 low-risk and 9 intermediate-risk patients according to FISH results showed complex karyotype by cytogenetics, and were classified into high-risk cytogenetic subgroup. Conclusion DSP30 and IL-2 are effective in improving the detection rate of CA in CLL patients (60.71%) and CA is more effective to detect complex karyotype. However, FISH had a higher overall abnormality detection rate (87.06%) than CA, especially for 13q-. The combination of CA and FISH not only enhanced the detection rate of clonal aberrations to 91.76%, but also provided more precise prognosis stratification for CLL patients, thus to provide more information for clinical implication.

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 CMML 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).
Chromosome Banding ; Chromosomes, Human, Pair 12 ; Chromosomes, Human, Pair 22 ; Cytogenetics ; Humans ; In Situ Hybridization, Fluorescence ; Leukemia, Myeloid ; genetics ; Oncogene Proteins, Fusion ; Translocation, Genetic

Objective To study the feasibility of clinical application of an individualized customized material. Methods Five batches of individualized customized materials were randomly selected, from which 10 cm × 11 cm samples were intercepted for experimental analysis. Among them, 10 cm × 10 cm materials were selected to perform dosimetric analysis and HU change analysis before and after irradiation with a radiotherapy dose for breast cancer of 50 Gy as the irradiation basis. The center Point 1 on the lower surface of the individualized material and the center Point 2 of the solid water volume were selected for dosimetric analysis before and after the sample is irradiated. After reaching a sufficient amount of irradiation, the 1 cm × 10 cm materials intercepted in the center position and the remaining 1 cm × 10 cm materials after the first sampling were sent to the material science laboratory for analysis of physical properties of density, viscosity, hardness, and tear strength. Results In the comparative analysis of HU values before and after exposure, after receiving 50 Gy dose irradiation, the difference rate of HU value was 5.252%, which was close to the expected 5% difference rate in clinical medicine. In the dosimetric analysis of Point 1 and Point 2, the dose in the irradiated samples was significantly higher than that in the unirradiated samples; the dose in Point 1 increased by 3.742%, and the dose in Point 2 increased by 2.039%. Before and after irradiation, except for the physical density which showed a significant difference, there was no significant difference in viscosity, hardness, and tear strength. Conclusion The individualized customized material can meet the requirements of routine clinical medicine.