2.A modified cytogenetic study for multiple myeloma.
You-wen QIN ; Dao-lin WEI ; Chun WANG ; Shi-ke YAN ; Yan-rong GAO ; Qi CAI
Chinese Journal of Medical Genetics 2006;23(2):213-215
OBJECTIVETo evaluate the effect of modified culture method used to cytogenetic analysis and the clinically significance of chromosomal abnormalities to multiple myeloma (MM).
METHODSMononuclear cells were isolated from bone marrow aspirate of 20 MM patients; and then cultured for 3 days without any cytokines, and 6 days in the presence of IL-6 (10 ng/mL) and GM-CSF (30 ng/mL) before RHG banding analysis; the remained part of aspirates were treated directly. Eight cases of iron deficiency anemia were taken as control.
RESULTSThe experiment was failure in 2 cases because of blood clot, and another 2 cases could be analyzed only by direct method due to inadequate cells. The karyotype abnormalities were found from 4 cases of 16 available patients. Of them, three cases had complex karyotypes. The abnormalities were detected after 6 days culture with addition of cytokines. No abnormalities were detected from those groups of directly analysis and 3 day culture. Meantime, the clinical data showed that the patients with cytogenetic abnormalities were in stage III, and had a high percentage of MM cells (25%-56%) in their bone marrow, and also poor responses to prior chemotherapy. No cytogenetic abnormalities were found from control individuals in all groups.
CONCLUSIONExtended culture in the presence of cytokines could improve the efficiency of cytogenetic analysis to MM. Complex karyotype was common cytogenetic abnormalities in MM patients with poor response to chemotherapy.
Aged ; Chromosome Aberrations ; Cytogenetic Analysis ; Cytokines ; metabolism ; Female ; Humans ; Karyotyping ; Male ; Middle Aged ; Multiple Myeloma ; genetics ; pathology
3.Inhibitory effect of RNA interference on chronic myeloid leukemia bcr/abl oncogene expression.
Xiao-xia MA ; Chun WANG ; Ju WEI ; You-wen QIN ; Shi-ke YAN ; Yan-rong GAO ; Qi CAI
Chinese Journal of Hematology 2005;26(6):359-362
OBJECTIVETo investigate the inhibitory effect of RNA interference on chronic myeloid leukemia (CML) bcr/abl oncogene expression.
METHODSThe small interference RNAs (siRNAs) were synthesized in vitro. K562 cells stably expressing bcr/abl gene were transfected with the siRNA by electroporation, both the non-transfected cells and non-specific siRNAs transfected cells were taken as controls. The enhanced green fluorescent protein (EGFP) plasmid was used as positive control and the transfection efficiency was detected by flow cytometry. Inhibitory effect of siRNAs was demonstrated by real-time quantitative RT-PCR and Western blots. Cell proliferation was measured by MTT assay and apoptosis by Annexin V-FITC assay.
RESULTSThe transfection efficiency was about 70%. The synthesized siRNAs inhibited CML bcr/abl oncogene expression at both mRNA and protein levels. siRNAs could inhibit K562 cell proliferation to 47% and 56% at 24 h and 48 h after transfection, respectively, and induce cell apoptosis from 1.00% in control group to 15.05% and 19.4% at 24 h and 48 h respectively.
CONCLUSIONAt the cell level, inhibition of CML bcr/abl oncogene expression by chemically synthesized siRNAs provides the new method for anti-leukemia study.
Apoptosis ; genetics ; Cell Proliferation ; Fusion Proteins, bcr-abl ; genetics ; Humans ; K562 Cells ; RNA, Small Interfering ; Transfection
4.Detection of cytogenetic abnormalities involving chromosomes 5,7 and 8 in myelodysplastic syndromes with fluorescence in situ hybridization and its clinical significance.
Yu CAI ; You-wen QIN ; Chun WANG ; Juan YANG ; Shi-ke YAN
Chinese Journal of Hematology 2007;28(1):6-10
OBJECTIVETo identify the abnormal karyotypes by fluorescence in situ hybridization (FISH) and explore prognostic implications in patients with myelodysplastic syndromes (MDS).
METHODSFISH was used to detect the frequently occurring chromosome abnormalities (-5/5q, +8, -7/7q-) in 37 MDS cases. SPSS 11.5 software and correlation analysis were used to analyze the relativity among the abnormal chromosomes, the prognosis and the disease conversion in 37 MDS patients.
RESULTSKaryotype abnormalities were found in 21 (56.8%) of 37 cases, among which 6 (16.2%) were complex karyotypes, 9 (24.3%) +8, 2(5.4%) -5/5q-, 2(5.4%) -7/7q-. In the median time of follow-up of 12 months, 12 cases transformed into acute leukemia. Complex karyotypes were significantly associated with the poor prognosis and leukemia transformation. + 8 and -7/7q- abnormalities were correlated with the death.
CONCLUSIONSFISH was more sensitive than conventional cytogenetics for detecting mini-clonal abnormality. There are some differences in abnormal karyotypes between patients in China and the western countries. Multi-probes used in cytogenetic detections may predict the patient' s prognosis more accurately. The higher proportion of abnormal karyotypes the poorer prognosis.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Chromosome Aberrations ; Chromosomes, Human, Pair 5 ; genetics ; Chromosomes, Human, Pair 7 ; genetics ; Chromosomes, Human, Pair 8 ; genetics ; Cytogenetic Analysis ; Female ; Humans ; In Situ Hybridization, Fluorescence ; Karyotyping ; Male ; Middle Aged ; Myelodysplastic Syndromes ; genetics
5.MRI Features and Site-specific Factors of Ischemic Changes in White Matter: A Retrospective Study
You-Ping ZHANG ; Na LIU ; Kai-Yan LIU ; Chao PAN ; Xuan CAI ; Shi-Qi YANG ; Zhou-Ping TANG ; Sha-Bei XU
Journal of Huazhong University of Science and Technology (Medical Sciences) 2018;38(2):318-323
Brain magnetic resonance imaging (MRI) of the elderly often reveals white matter changes (WMCs) with substantial variability across individuals.Our study was designed to explore MRI features and site-specific factors of ischemic WMCs.Clinical data of consecutive patients diagnosed with ischemic cerebral vascular disease who had undergone brain MRI were collected and analyzed.Multi-logistic regression analysis comparing patients with mild versus severe WMCs was performed to detect independent associations.Analyses of variance (ANOVAs) were used to detect regionally specific differences in lesions.We found that lesion distribution differed significantly across five cerebral areas,with lesions being predominant in the frontal lobe and parieto-occipital area.To explore WMCs risk factors,after adjusting for gender,diabetes mellitus,and hypertension,only age (P<0.01),creatinine (P=0.01),alkaline phosphatase (ALP) (P=0.01) and low-density lipoprotein cholesterol (LDL-C) (P=0.03) were found to be independently associated with severe WMCs.Age (P<0.001) was strongly associated with WMCs in the frontal lobe while hypertension was independently related to lesions in the basal ganglia (P=0.048) or infratentorial area (P=0.016).In conclusion,MRI of WMCs showed that ischemic WMCs occurred mostly in the frontal lobe and parieto-occipital area.The infratentorial area was least affected by WMCs.Typically,age-related WMCs were observed in the frontal lobes,while hypertension-related WMCs tended to occur in the basal ganglia and infratentorial area.
6.Clinical features and correlative factors of senile coronary heart disease accompanied with depression
Ji-ying LIANG ; Cai-you HU ; Wen-jing PAN ; Zeping LV ; Xiaoyan DUAN ; Wen SHI ; Weihong SU ; Chuyu YANG ; Xiuta YANG
Chinese Journal of Rehabilitation Theory and Practice 2004;10(10):616-617
ObjectiveTo investigate clinical features, correlative factors and prognosis of senile coronary heart disease accompanied with depression.MethodsFifty two senile patients of coronary hart disease with depression were selected for the observation group, sixty cases without depression as control group. Clinical features, family conditions, society factors and Holter report (supervision) of two groups were analyzed.ResultsClinical features such as severe dyssomnia, anxiety agitation, emotion depressing, loss of interest, feeling waste and sense of guilty in patients of observation group were significant different with those in control group. The rate of family harmoniousness and society supporting between patients of two groups were significantly different. The incidence of ventricular arrhythmias, atrial arrhythmia and myocardial ischemia changes on ST-T in the Holter reports of patients in observation group were higher than those in control group (P<0.05 or P<0.01). The incidence of cardiac sudden death of patients in observation group was also higher than patients in control group (P<0.05).ConclusionClinical symptoms such as severs dyssomnia, anxiety agitation, emotion depressing are easily occurred in senile coronary heart disease patients with depression. Patients with depression have higher incidence of cardiac sudden death, having better relationship and society support are advantageous to prevent and reduce the depression in senile coronary heart disease patients.
7.CT-guided puncture for accurate radiofrequency thermocoagulation through the pterygopalatine fossa approach in treatment of maxillary neuralgia
You-Cai SHI ; Yan-Mei WANG ; Tao LIU ; Bin-Hua XIU ; Shao-Ming LIU
Chinese Journal of Neuromedicine 2012;11(4):380-382
Objective To study the therapeutic effect of CT-guided puncture for accurate radiofrequency thermocoagulation through the pteroygopalatine fossa approach in the treatment of maxillary neuralgia. Methods From May 2005 to March 2011, 18 patients with maxillary neuralgia underwent CT-guided puncture through the pteroygopalatine fossa approach for accurate radiofrequency thermocoagulation. Axial CT scanning parallel to the middle cranial fossa base was conducted for the patients to define that the puncture needle was within the pteroygopalatine fossa before radiofrequency thermocoagulation being performed at 85℃ for 60 seconds. The thermocoagulation was repeated once until anesthesia covered the innervation area of the superior alveolar nerve. Results Follow-up,ranging from 6 to 22 months (mean,10 months),showed successful puncture in all patients.According to the Heft-Parker' visual analog scale, 15 cases (83.3%) were rated as excellent and 2 as good (11.1%).There were no intraoperative or postoperative major complications in the patients. Conclusions Since CT-guided puncture through the pteroygopalatine fossa approach can safely ensure accurate radiofrequency thermocoagulation for treatment of maxillary neuralgia, it has a high therapeutic efficacy for the patients.It is also a minimally invasive procedure that can be easily and safely performed in clinic.
8.Evaluation of the subsets of lymphocytes and their activated status in patients with myelodysplastic syndrome.
Jun YANG ; Chun WANG ; Kuang-Cheng XIE ; Shi-Ke YAN ; Yan-Rong GAO ; Qi CAI ; You-Wen QIN ; Li-Ping WAN ; Yu CAI
Journal of Experimental Hematology 2006;14(4):708-713
This study was purposed to investigate the clinical significance of the amount and activated status of T cell subsets, B cells, NK cells in peripheral blood from patients with myelodysplastic syndrome (MDS). The proportion of T cells, B cells, NK cells in peripheral blood from 30 patients with MDS and their surface activation markers of CD28, CD45RA, CD45RO, CD69, HLA-DR were analyzed by flow cytometry. Twenty-two patients were in the low risk group (RA + RAS) while eight patients were in the high risk group (RAEB + RAEBT). The result showed that the amounts of T cells (CD3+ cells) in peripheral blood from patients with MDS were lower than those in control group. The amounts of naive CD4+ cells (CD4+ CD45RA+ cells) in MDS patients were lower than those in control. The expression rates of early activation marker (CD69) and late activation marker (HLA-DR) on CD3+ cells in MDS patients were significantly higher than those in control. The abnormalities of the immunologically competent cells were mainly observed in the low risk group (RA + RAS), and were characterized by the high expression rates of CD69+ and HLA-DR+ on CD3+ cells, the decrease of B cell amounts. The amount abnormalities of T cell subsets were mainly observed in high risk group (RAEB + RAEBT), and were characterized by the decrease of CD3+ cells and CD3+ CD4+ CD8- cells (Th cells) amounts without high expression of the CD69 and HLA-DR, the decrease of NK cells amounts. It is concluded that there are the abnormalities of T cell subsets and function in the patients with MDS and may change with disease progression, so the measurement of amount and activated status of T cell subsets in peripheral blood from MDS patients can have predictive role for diagnosis of disease progression and guide of therapy.
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9.Inhibitory effect of Rnai on AML1 -ETO fusion gene expression in leukemia cells.
Ju WEI ; Su LI ; Chun WANG ; You-Wen QIN ; Xiao-Xia MA ; Kuang-Cheng XIE ; Shi-Ke YAN ; Yan-Rong GAO ; Qi CAI
Chinese Journal of Hematology 2008;29(9):607-610
OBJECTIVEBy inhibiting AML1 -ETO fusion gene expression in Kasumi-1 cells with RNAi, to investigate the changes in cell proliferation and cell cycle.
METHODSThe small interference RNAs (siRNAs) specifically targeting the AML1 -ETO fusion gene were synthesized in vitro and transfected into Kasumi-1 cells by electroporation, the non-specific siRNAs transfected cells were taken as control. EGFP plasmid was transfected into Kasumi-1 cell and the transfection efficiency was detected by FCM. Inhibitory effect of siRNAs were detected by real-time RT-PCR and Western blots. Cell proliferation was measured by CCK-8 assay. DNA content was detected by PI assay.
RESULTSThe transfection efficiency was 44.5%. The AML1 -ETO specific siRNAs inhibited AML1 -ETO expression at both mRNA and protein levels. The cell proliferation rate in siRNAs treated group was lower than that in control group 72 h after transfection [(47.90 +/- 0.02)% vs (66.90 +/- 0.08)% , P < 0.05]. The cell cycle was blocked at G1 phase 72 h after siRNAs treatment, the cell proportion in G1 phase being 38.3% and 31.6% in control group, while in G2/M phase being 1.8% and 2.4% respectively.
CONCLUSIONSThe synthesized siRNAs can inhibit AML1 -ETO fusion gene expression. AML1 -ETO specific siRNA induced the decline of AML1 -ETO fusion protein in Kasumi-1 cell, and then caused the cell cycle blocked in G1 stage and eventually inhibited the cell proliferation.
Cell Cycle ; genetics ; Cell Line, Tumor ; Cell Proliferation ; Core Binding Factor Alpha 2 Subunit ; genetics ; metabolism ; Humans ; Leukemia ; genetics ; metabolism ; pathology ; Oncogene Proteins, Fusion ; genetics ; metabolism ; RNA Interference ; RUNX1 Translocation Partner 1 Protein ; Transfection
10.Allogeneic hematopoietic stem cell transplantation following reduced intensity conditioning regimen for treatment of refractory leukemia.
Jie-Ling JIANG ; Shi-Ke YAN ; Juan YANG ; Yu CAI ; Li-Ping WAN ; You-Wen QIN ; Chun WANG
Chinese Journal of Hematology 2008;29(8):517-521
OBJECTIVETo evaluate the efficacy of allogeneic hematopoietic stem cell transplantation (allo-HSCT) following reduced intensity conditioning (RIC) regimen for treatment of refractory leukemia.
METHODSTwenty patients with refractory leukemia received allo-HSCT following RIC regimen consisting of fludarabine plus small or moderate dose total body irradiation (TBI). Graft versus host disease (GVHD) prophylaxis was CsA plus mycophenolate mofetil (MMF) or short-term MTX, or these three drugs combination; CD25 monoclone antibody(McAb) and ATG were also used in some of the patients.
RESULTSSeventeen patients engrafted successfully, the median time for ANC > 0.5 x 10(9)/L was 13 (11 - 17) days, and for BPC > 50 x 10(9)/L 19 (12 -42) days. Detected by short tandem repeat (STR)-PCR, complete donor chimerism was confirmed in 16 patients with a median of 14 (7 -35) days. The incidence of acute and chronic GVHD was 47.1% (8/17) and 38.5% (5/13) respectively. The transplant related mortality (TRM) was 25.0% (5/20), mainly from graft failure, intracranial hemorrhage and severe infection. Up to now, 7 patients relapsed and 9 were alive with leukemia free. The overall survival (OS) at 2 year was (35. 3 +/- 14.2)% for all patients and (52.5 +/- 18.6)% for acute non-lymphocytic leukemia (ANLL) patients.
CONCLUSIONAllo-HSCT following fludarabine and TBI based RIC regimen can be used for treatment of refractory leukemia with well tolerance and low TRM and there is a better prognosis for ANLL patients than that for acute lymphocytic leukemia patients.
Adolescent ; Adult ; Child ; Female ; Graft vs Host Disease ; prevention & control ; Hematopoietic Stem Cell Transplantation ; methods ; Humans ; Leukemia ; therapy ; Male ; Middle Aged ; Retrospective Studies ; Transplantation Conditioning ; methods ; Treatment Outcome ; Young Adult