Analysis of ASXL1 gene variant in patients with myelodysplastic syndrome / 中华医学遗传学杂志

OBJECTIVE@#To detect ASXL1 gene variants among patients with myelodysplastic syndrome (MDS) and explore their correlation with variants of other genes and clinical features of patients.@*METHODS@#For 149 patients with MDS, genomic DNA was amplified by PCR and subject to direct sequencing to identify variants of ASXL1, U2AF1, SF3B1, DNMT3A, TET2, IDH1/2, NPM1, FLT3-ITD and C-KIT genes.@*RESULTS@#ASXL1 variants were found among 37 patients (24.8%). Other commonly mutated genes included U2AF1 (22.8%), TET2 (11.4%), DNMT3A (9.4%), NPM1 (8.1%) and SF3B1 (6.0%). The frequency of concurrent U2AF1 and TET2 variants among patients with ASXL1 variants was slightly higher than that of wild-type patients. No significant difference was found in median age, MDS subtype, karyotype, peripheral leukocytes, hemoglobin, platelet levels, and bone marrow blast counts between the ASXL1-variant and the wild-type groups (P> 0.05). Twenty-nine patients harboring ASXL1 variants were followed up, 37.9% progressed to acute myeloid leukemia (AML). The rate of transformation in ASXL1-variant group was significantly higher than the wild-type group (37.9% vs. 14.1%, P< 0.01).@*CONCLUSION@#ASXL1 showed a high frequency of variant among MDS patients, which was frequently accompanied with U2AF1 and TET2 variants. Compared with the wild type group, patients with ASXL1 variants were more likely to progress to AML.

Clinical feature and cytogenetic analysis of 28 patients with bone marrow invasion non-Hodgkin's lymphoma / 中华医学遗传学杂志

OBJECTIVE@#To study the correlation of hematomorphology, bone marrow cytogenetics and clinical biochemical parameters with the prognosis of non-Hodgkin's lymphoma with bone marrow invasion.@*METHODS@#Morphological analysis of bone marrow cells was performed by routine bone marrow puncture.Chromosome samples were prepared by short-term bone marrow culture. Karyotype analysis was carried out by R-banding in 28 patients. P53 gene was detected by fluorescence in situ hybridization (FISH). Serum lactate dehydrogenase (LDH) of all patients was determined and compared.@*RESULTS@#In all patients, bone marrow morphology showed invasion of lymphoma. Chromosome analysis revealed abnormal karyotypes in 19 cases, which yielded an incidence of 67.85%. The proportion of lymphoma cells in bone marrow among those with an abnormal karyotype was much higher than those with a normal karyotype (60.2% vs. 33.5%, P<0.05). FISH assay showed that 9 (32.14%) patients had P53 gene deletion. And the deletion was much more common among those with an abnormal karyotype (42.11% vs. 11.11%, P<0.05). The serum LDH level in patients with an abnormal karyotype was significantly higher compared with whose with a normal karyotype (1464.37 U/L vs. 294.33 U/L, P<0.05).@*CONCLUSION@#Patients with abnormal karyotypes have a higher rate of P53 gene deletion, and their LDH level is significantly higher than those with a normal karyotype, which predicted a relatively poor prognosis.

Characterizing the molecular cytogenetics in acute monocytic leukemia / 中华医学遗传学杂志

OBJECTIVE@#To characterize the molecular genetics of 81 patients with acute monocytic leukemia (AML).@*METHODS@#Fluorescence in situ hybridization (FISH) was employed to detect MLL gene rearrangements. Combined mutations of 17 genes were detected by DNA-based PCR and Sanger sequencing.@*RESULTS@#Sixty seven patients were found to harbor at least one mutation. The most commonly mutated gene was NPM1 (n=18), which was followed by FLT3-ITD (n=16), NRAS (n=16), DNMT3A (n=15), TET2 (n=12), RUNX1 (n=11) and KRAS (n=9). Based on the functions of mutated genes, the most frequently involved genes were those involved in DNA methylation (38.27%), tyrosine kinase receptor signaling (32.1%), transcription regulation (28.4%), and RAS pathway (24.7%). Single gene mutation predominated in patient with cytogenetic abnormalities, while coexistence of 2 mutations have predominated in patient with normal cytogenetic findings. Stratified by cytogenetic findings, patients with single gene mutations (intermediate-risk group) had significantly higher complete remission (CR) rates than those with ≥2 gene mutations (unfavorable-risk group) (91.7% vs. 57.6% , 87.5% vs. 25.0%, P =0.0319, 0.0117, respectively).@*CONCLUSION@#Over 80% of AML patients were found to harbor at least one mutation. Their clinical phenotype and prognosis may be impacted by the synergy of MLL gene rearrangement and multiple mutations. For patients under the same risk stratification, the number of mutations is reversely correlated with the CR rate.

Clinical and genetic features of a patient with myeloid neoplasm in association with PDGFRA and EVI1 gene rearrangements / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>Todelineate the clinical and genetic features of a patient with myeloproliferative neoplasm (MPN) in association with PDGFRA and EVI1 genes rearrangements.</p><p><b>METHODS</b>Clinical data of the patient was collected. Conventional cytogenetics, fluorescence in situ hybridization (FISH) and nested PCR were carried out for the patient.</p><p><b>RESULTS</b>The patient has featured recurrent rash, joint pain, and intermittent fever. Laboratory tests showed hyperleukocytosis and marked eosinophilia. Physical examination revealed splenomegaly. His karyotype was 46,XY,t(3;5)(q26;q15)[6]/46,XY[10]. FISH assay showed that both PDGFRA and EVI1 genes were rearranged. Molecular studies of the mRNA suggested that there was a in-frame fusion between exon 12 of the PDGFRA gene and exon 9 of the FIP1L1 gene. Imatinib was initiated at a dosage of 200 mg, and after 10 months, the signal of the FIP1L1-PDGFRA fusion gene was undetectable in bone marrow sample. However, the expression of EVI1 mRNA was stable, with no significant difference found between the patient and 10 healthy controls.</p><p><b>CONCLUSION</b>MPN in association with PDGFRA and EVI1 genes rearrangements have unique clinical and genetic features. Genetic testing is helpful for early diagnosis. Imatinib may be effective for the treatment.</p>

Correlation of cytogenetic changes with VEGF and TRacp-5b levels among 60 elderly patients with multiple myeloma / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To assess the correlation of cytogenetic changes with serum vascular endothelial growth factor (VEGF) and serum tartrate resistant acid phosphatase (TRacp-5b) levels among elderly patients with multiple myeloma (MM).</p><p><b>METHODS</b>Chromosomal changes were analyzed with a modified culturing method in the presence of IL-6. Serum levels of VEGF and TRacp-5b were determined with enzyme-linked immunosorbent assays (ELISA).</p><p><b>RESULTS</b>Among the 60 MM patients, chromosomal abnormalities were found in 27 cases, including 22 with numerical abnormalities and 15 with structural abnormalities. Many patients had both numerical and structural abnormalities. For 33 patients with a normal karyotype, the levels of VEGF and TRacp-5b were 117.35 ± 55.26 pg/mL and 4.15 ± 2.15 U/L, respectively, while for 27 patients with an abnormal karyotype, the levels of VEGF and TRacp-5b were 190.26 ± 85.74 pg/ml and 5.96 ± 2.24 U/L, respectively. The difference between the two groups was significant (P<0.05).</p><p><b>CONCLUSION</b>Compared with MM patients with a normal karyotype, the levels of VEGF and TRacp-5b are higher in those with cytogenetic abnormalities.</p>

CALR gene mutation detection and clinical observation of 150 essential thrombocythemia patients / 中华血液学杂志

<p><b>OBJECTIVE</b>To explore the prevalence of CARL gene mutations and the mutation types in patients with essential thrombocythemia (ET), and to compare the patients clinical characteristics of CALR mutation with JAK2 V617F, MPL W515K mutation patients and triple negative group.</p><p><b>METHODS</b>The mutations of CALR gene at extron 9 and MPL W515K in 150 ET patients were detected by PCR amplification followed by direct sequencing of genomic DNA, the JAK2 V617F mutation by using allele specific PCR.</p><p><b>RESULTS</b>(1)The CALR mutations were found in 38 patients (25.3%) of 150 ET patients. A total of 4 types of CALR mutations were identified (type Ic.1092_1143del52bp, n=17; type II c.1154_1155insTTGTC, n=16; type III c.1094_1139del46bp, n=4; type IV c.1103_1136del34bp, n=1). (2)The incidence of JAK2 V617F and MPL W515K was 61.3% (92/150) and 2.7% (4/150), respectively. The frequency of CALR mutation was 70.4% (38/54) in 54 ET patients without JAK2 V617F and MPL W515K mutations. The co-occurrence of any two kinds of gene mutations was not detected. (3)The hemoglobin level and leukocyte counts of patients with CARL mutations were significantly lower than that in patients with JAK2 V617F mutations (P<0.05). The median age of patients with CALR mutation was significantly higher than that of triple negative patients (59 years vs 29.5 years, P<0.01). Cytogenetic analysis was performed in 147 patients, and there were 4 abnormal karyotype cases. CALR mutation incidence was significantly higher in abnormal karyotype cases than that in normal ones (75% vs 24.5%, P=0.019).</p><p><b>CONCLUSION</b>The incidence of CALR mutations is high in ET patients without JAK2 V617F and MPL W515K mutations, and is associated with abnormal karyotype. CARL-mutated cases showed a significantly lower leucocyte and hemoglobin levels compared with JAK2 V617F mutated cases.</p>

Clinical feature and cytogenetic analysis of 80 patients with acute monocytic leukemia / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To explore the relationship between cytogenetic and clinical features and prognosis for patients with acute monocytic leukemia (M5 type).</p><p><b>METHODS</b>Chromosome samples were prepared by direct culture of bone marrow for 24 hours. Karyotypes of 80 patients with M5 were analyzed by R banding. Rearrangement of MLL gene and deletion of P53 gene were detected by fluorescence in situ hybridization (FISH).</p><p><b>RESULTS</b>Forty-three patients (53.75%) were found to have abnormal karyotypes, which included 23 patients with abnormalities of chromosome 11 and 20 with other chromosomal abnormalities. Twenty-four patients had MLL gene rearrangements and 17 had P53 gene deletion.</p><p><b>CONCLUSION</b>11q23 has been the most common chromosomal abnormality among patients with M5, which is associated with poor prognosis. The frequency of P53 gene deletion in patients with genetic abnormalities were significantly higher than those with normal karyotypes (P < 0.05).</p>

Preliminary studies on the serum IL-17 and TGF-β levels and their correlations with tumor incidence and progression in colorec-tal cancer patients / 中国肿瘤临床

Objective: This work aimed to investigate the relationships of the serum levels of IL-17 and TGF-β with the carcinogenesis and progression of colorectal cancer (CRC), as well as the clinical significance of these serum levels. Methods: Data of 30 healthy subjects, 59 patients with simple CRC and 44 CRC patients with postoperative (post-op) metastasis were recruited in this study. The patients were respectively divided into group A (30 healthy subjects as the control group), group B (59 CRC patients without distant metastasis after surgery), and group C (44 CRC patients with post-op metastasis). The patients in each group had a mean age of 53.8 ± 20.8, 62.0 ± 11.8, and 64.0 ± 15.7 years, respectively. All patients were confirmed by pathological diagnosis. The serum levels of IL-17 and TGF-β were measured by enzyme-linked immunosorbent assay. All quantitative data were analyzed using SPSS 13.0. Results: The IL-17 serum level was significantly higher in groups B and C than in group A. The preoperative (pre-op) serum level of IL-17 was significantly higher than the post-op serum level in group B (P0.05). However, the TGF-β serum level in group C was significantly higher than that in groups A and B (P<0.05). No significant correlation was observed in the serum level IL-17 or TGF-β between colon and rectum cancers in groups B and C. Conclusion: The serum level of IL-17 is significantly correlated with that of CRC. The serum level IL-17 increases with the aggravation of CRC and increased tumor burden. A strong correlation exists between the serum level of TGF-β and metastasis of CRC. Cytokine IL-17 and TGF-β may play an important role in the progression and metastasis of CRC.

Association between urinary cadmium and clinicopathological characteristics of breast cancer / 中华肿瘤杂志

<p><b>OBJECTIVE</b>The aim of this study was to investigate the association between urinary cadmium and clinicopathological characteristics of breast cancer.</p><p><b>METHODS</b>The clinicopathological characteristics of 240 patients with breast cancer were obtained and urine specimens were collected from October 2009 to July 2010. The concentration of urinary cadmium was determined by inductively coupled plasma mass spectrometry (ICP-MS). χ(2) test and Wilcoxon rank sum test were used to analyze whether urinary cadmium is associated with clinicopathological characteristics of breast cancer.</p><p><b>RESULTS</b>The median concentration of urine cadmium of 240 patients was 1.99 µg/g (25th percentile, 1.32 µg/g; 75th percentile, 2.88 µg/g). HER-2 positive rate, regional/distant metastasis rate, and advanced stage rate in patients with the highest tertile of cadmium concentration were significantly higher than those in the patients with second and lowest Cd tertiles (P = 0.042, P = 0.028 and P = 0.017, respectively), and 28.2% vs. 16.5% for HER-2 and 47.2% vs. 32.0% for regional/distant metastasis, respectively. There were still significant associations between urinary cadmium levels and these clinicopathological parameters after being adjusted in age by unconditional logistic regression model, respectively (P < 0.05).</p><p><b>CONCLUSIONS</b>The results of this study suggest that urinary cadmium levels are associated with the HER-2 status, regional/distant metastasis status and stages of breast cancer, respectively. Cadmium may induce highly aggressive breast cancer in humans.</p>

Interaction of body mass index and a polymorphism in gene of catalytic subunit of glutamate cysteine ligase on breast cancer risk among Chinese women / 中华流行病学杂志

Objective To investigate the interaction of body mass index (BMI) and a single nucleotide polymorphism (SNP,rs17883901) in catalytic subunit of glutamate-cysteine ligase (GCLC) on breast cancer risk.Methods A total of 839 women with incident breast cancer and 863 age-matched controls without cancer were recruited at the same period in three affiliated hospitals of Sun Yat-sen University in Guangzhou from October 2008 to June 2010.GCLC rs17883901 was genotyped by MALDI-TOF-MS.Binary unconditional logistic regression was applied to calculate odds ratios and 95％ confidence intervals.Results The difference of present BMI and BMI at age 20 was not statistically significant between cases and controls,either as the genotypes of GCLC.No association was found between BMI at present and premenopausal or postmenopausal breast cancer risk.But we found that women who had a BMI at age 20 of 18.5 to 22.9 had a marginally decreased risk of premenopausal breast cancer [OR and 95％CI:0.69 (0.48,1.00)].Among women with CT/TT genotypes,whose present BMI was greater than 25 had a increased risk [OR and 95％CI:1.91 (1.09,3.36)] of breast cancer and a decreased risk [OR and 95％CI:0.56(0.31,0.99)] with a BMI at age 20 of 18.5 to 22.9.There was a interaction between GCLC gene (rs17883901)and BMI at present in breast cancer risk (P=0.043),which was not found between rs17883901 and BMI at age 20.Conclusion Our findings indicate BMI at age 20 may be a protective factor of premenopausal breast cancer,while no association appears between GCLC (rs17883901) and breast cancer.Obesity at present may significantly increase the risk of breast cancer among women with CT/TT genotypes of GCLC (rs17883901).

Prognostic value of P53 aberrations in diffuse large B-cell lymphoma / 中国实验血液学杂志

The aim of this study was to analyze the P53 aberrations in diffuse large B-cell lymphoma (DLBCL) and its prognostic value. Using fluorescence in situ hybridization (FISH), the P53 gene was detected in paraffin-embedded tonsil tissues from 50 cases of DLBCL, while all the peripheral blood of patients was collected for detecting P53 protein in serum by using enzyme-linked immunosorbent assay (ELISA). The relationships between P53 gene and protein expression and the prognosis of patients with DLBCL were analyzed. The results showed that 21 out of 50 cases had P53 gene deletion, serum P53 protein content was (176.25 ± 61.25) pg/ml, which was higher than that in normal controls. The Cox model likelihood ratio test found that abnormal P53 could be used as independent prognostic factor in DLBCL patients. The risk of death in patients with P53 gene deletion were more than that in patients with P53 gene normal. The patients with histology P53 gene deletion detected by FISH also had relatively high level of mutant P53 protein in serum. It is concluded that P53 aberrations in patients with DLBCL can be used as an independent prognostic factor, the early detection of P53 in DLBCL patients helps to determine their prognosis accurately.

Analysis of IDH1 and IDH2 mutations in patients with acute myeloid leukemia / 中华血液学杂志

<p><b>OBJECTIVE</b>To explore the prevalence of IDH gene (IDH1 and IDH2) mutations, types of mutations in patients with acute myeloid leukemia (AML), correlation with the internal tandem duplication(ITD) mutation of FLT3 gene, NPM1 gene mutation and some clinical characteristics.</p><p><b>METHODS</b>The mutations of IDH1 and IDH2 gene at exon 4, NPM1 gene at exon 12 and FLT3-ITD at exon 14 and 15 in 163 newly diagnosed AML patients were detected by PCR amplification followed by direct sequencing of genomic DNA.</p><p><b>RESULTS</b>(1) IDH mutations were found in 25 patients (25/163), and all were heterozygous, of which IDH1 in 7 patients (4.29%) and IDH2 in 18 (11.04%). A total of 4 types of IDH1 mutations were identified (c.395G→A, p.R132H, n = 4; c.394C→A, p.R132S, n = 1; c.394C→G, p.R132G, n = 1; c.315C→T, n = 1). The IDH1 mutation caused substitutions of residue R132 except for one (c.315C→T). All IDH2 mutations caused changes of R140 (c.419G→A, p.R140Q, n = 18). The incidence of IDH2 mutation was significantly higher than that of IDH1 mutation (11.0% v 4.3%, P = 0.022). Both IDH1 and IDH2 mutation were detected in one patient, while IDH1 was synonymous substitution (c.315C→T). IDH-mutated cases showed a significantly higher frequency of concurrent FLT3-ITD mutation compared with wildtype cases (34.6% vs 11.9%, P = 0.003), so did IDH mutations concurrent NPM1 mutation vs NPM1 wildtype (28.1% vs 12.7%, P = 0.033), of which the frequency of concurrent NPM1 and FLT-ITD mutations cases with the IDH mutation was significantly higher than that of NPM1 and FLT-ITD negative (45.5% vs 11.7%, P = 0.002). IDH mutation incidence was significantly higher in normal karyotype cases than in abnormal ones (20.5% vs 5.8%, P = 0.020). Patients with IDH mutations were significantly older than wildtype patients(P < 0.001), whereas, there were no statistically significant differences in gender, peripheral blood (PB) count at diagnosis between two groups.</p><p><b>CONCLUSIONS</b>The incidence of IDH mutation is higher in patients with de novo AMLs, of which IDH2 mutation more frequently, and the patients associated with older age, normal karyotype at diagnosis. IDH mutation has a strong association with NPM1 and FLT3-ITD mutations, suggesting that IDH mutation has synergistic effect with the latter gene on leukemogenesis.</p>

Prognostic value of t(11; 18) (q21; q21) for gastric mucosa-associated lymphoid tissue lymphoma / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate the prognostic value of t(11; 18) (q21; q21) in gastric mucosa-associated lymphoid tissue lymphoma.</p><p><b>METHODS</b>A cohort of thirty-six gastric mucosa-associated lymphoid tissue lymphoma patients who were pathologically identify diagnosis from January 1994 to June 2004 were followed up retrospectively and studied using fluorescence in situ hybridization(FISH) technique to detect t(11; 18) (q21; q21) chromosomal translocation on preservative paraffin specimen.</p><p><b>RESULTS</b>Among thirty-six patients, fifteen (41.67%) were positive for t (11; 18) (q21; q21). All but one were followed up to March 2010, general median survival time (MST) was 87 months. The MST were 43 and 130 months for t(11; 18) positive and negative patients, respectively. The MST between these two groups was notably different (chi-square=29.57, P< 0.01).</p><p><b>CONCLUSION</b>t(11; 18) (q21; q21) is important prognostic factor for gastric mucosa-associated lymphoid tissue lymphoma.</p>

Clinical and cytogenetic analyses of 45 adult patients with acute lymphoblastic leukemia / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To analyze the correlation between clinical features and cytogenetic finding of 45 adult patients with acute lymphoblastic leukemia (ALL), and to assess the value of chromosomal examination for the diagnosis and prognosis.</p><p><b>METHODS</b>Fluorescence in situ hybridization (FISH) was utilized for detecting the BCR/ABL fusion gene and P53 gene. Median survival time (MST) of patients was compared using Log-rank test.</p><p><b>RESULTS</b>Respectively, the MST of patients with white blood cell count (WBC) ≤30 × 10(9)/L, normal karyotype, or without a Philadelphia chromosome were significantly greater than those with WBC > 30 × 10(9)/L, abnormal karyotype or Philadelphia chromosome (P< 0.05).</p><p><b>CONCLUSION</b>WBC, karyotype abnormalities and presence of Philadelphia chromosome are independent factors for the prognosis of ALL in adult patients.</p>

Analysis of the non-Hodgkin lymphoma's clinic and blood marrow cytogenetics / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To analyze the relationship among cytogenetics, hematopathology, morphology, clinic tumor index and prognosis of the non-Hodgkin lymphoma(NHL) patients.</p><p><b>METHODS</b>Cytogenetics analysis of bone marrow cells was performed by direct method and 24 h culture method in 20 cases of NHL. Serous P53 protein was detected in all patients.</p><p><b>RESULTS</b>Out of the twenty NHL patients, eight had bone marrow infiltration, and nine were found with abnormal karyotype: three were chromosome number abnormalities, one was t(14,18)(q32;q21), one was 14q+, two were t(8;14)(q24;q32), two were t(8;14)(q24;q32) with additional changes of chromosome. The serous P53 protein of patients with abnormal karyotype was significantly higher than that with normal karyotype.</p><p><b>CONCLUSION</b>t(8;14) (q24;q32) is the most classic translocation in NHL, The patients with this karyotype have worse prognosis. The increase of mutant serous P53 relates to abnormal karyotype, and the patients with high mutant P53 level in serum also have worse prognosis.</p>

Analysis the clinic and cytogenetics of myelodysplastic syndrome / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To analysis the relationship among cytogenetics, morphology and prognosis of the myelodysplastic syndrome (MDS) patients.</p><p><b>METHODS</b>Cytogenetics analysis of bone marrow cells was performed by direct method and/or 24 h culture method. The karyotype was analysed by R banding technique.</p><p><b>RESULTS</b>Out of the 50 MDS patients, 22 were found with abnormal karyotype: two of them were +8, four of them were -7, four of them were 5q-, nine of them were 7q-, two of them were 20q- and one of them was 6q-. There are six patients had complex changes in chromosome.</p><p><b>CONCLUSION</b>5q-, -7, 7q- are the most classic translocation in the MDS. The patients with 5q- have better prognosis and the patients with -7, 7q- have worse prognosis. Cytogenetics is very important in the MDS's diagnosis, progress and prognosis.</p>