Synthesis and antisense properties of 2'β-F-/2'α-F-2'-deoxy-uridines modified oligonucleotides with 4'-C-(2-methoxyethoxy) substituent / 药学学报

Chemical modification is critical for the therapeutic applications of antisense oligonucleotides. Novel 4'-C-MOE and 2'-fluoro-modified monomer 2'-F-4'-C-MOE-araU and its epimeric 2'-F-4'-C-MOE-rU were synthesized from 2'-fluorinated arabinourine (2'-F-araU) and 2'-fluorouridine (2'-F-rU), respectively. Their phosphoramidites were synthesized and successfully incorporated into oligodeoxynucleotides. The mismatch discrimination ability of these unnatural monomers and their effect on thermal stability were evaluated in the context of dsDNA and DNA-RNA chimeras. The thermal denaturation studies showed that the incorporation of 2'-F-4'-C-MOE-araU led to enhanced binding affinity to complementary RNA strand and almost equivalent binding ability to complementary DNA, when compared with 2'-F-4'-C-MOE-rU and 2'-F-araU modified duplexes.Especially a C-H…F-C pseudohydrogen bond was supposed to contribute more binding affinity at uridine-purine steps, meanwhile, 2'-F-4'-C-MOE-araU had almost the same base discriminatory ability as uridine in dsDNA and DNA-RNA chimeras, while 2'-F-4'-C-MOE-rU was found to have only moderate RNA hybridization ability. However, 2'-F-4'-C-MOE-araU at 3'-end of oligonucleotide could not led to more nuclease hydrolytic stability than that with 2'-F-4'-C-MOE-rU modification. These results demonstrated the feasibility of C4'-MOE modification on 2'-F-ANA and the dramatic effects of the 2'-F substituent, which provides a new approach fo r further chemical modification of antisense drugs.

Shenfu Injection suppresses inflammation by targeting haptoglobin and pentraxin 3 in rats with chronic ischemic heart failure / 中国结合医学杂志

<p><b>OBJECTIVE</b>To investigate the regulatory effects of Shenfu Injection (SFI, ) on hemodynamic parameters and serum proteins in rats with post-infarction chronic heart failure (CHF).</p><p><b>METHODS</b>Forty-five healthy Wistar rats were randomized into three groups: sham, heart failure (model) and SFI group. The CHF was induced by left coronary artery ligation. Seven days after the surgical operation, animals in the sham group and the model group received saline (6.2 mL/kg/d), while animals in the SFI group received SFI (6.2 mL/kg d) intraperitoneally. Four weeks later, cardiac hemodynamic parameters were measured via the carotid route. The expression of serum proteins was analyzed by two-dimensional electrophoresis and matrix-assisted laser desorption/ionization time-of-flight mass spectrometer (MALDI-TOF MS).</p><p><b>RESULTS</b>Recording of hemodynamic parameters showed that left ventricular systolic pressure (LVSP), maximum rate of left ventricular pressure (+dp/dtmax) rise, and maximum rate of left ventricular pressure (-dp/dtmax) decrease, while the left ventricular end diastolic pressure (LVEDP) rose in the model group compared to those in the sham group (P <0.05). The results of the MALDI-TOF MS indicated that haptoglobin (HP), pentraxin 3 (PTX3) and alpha-1-antitrypsin were up-regulated, while serum albumin and 40S ribosomal protein were down-regulated in the model group (P <0.05). Compared with the model group, LVSP, +dp/dtmax and -dp/dtmax were higher, while LVEDP was lower in the SFI group (P<0.05). Expression levels of HP and PTX3 were lower than in the model group (P<0.05).</p><p><b>CONCLUSION</b>SFI could improve hemodynamic function and decrease inflammatory reactions in the pathophysiology of CHF. The serum proteins HP and PTX3 could be potential biomarkers for chronic ischemic heart failure, and they could also be the serum protein targets of SFI.</p>

Evaluation of respiratory dysfunction in a pig model of severe acute dichlorvos poisoning / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Respiratory failure is the main cause of death in acute organophosphorus pesticide poisoning. In this study, a pulse-induced contour cardiac output monitor was used to evaluate the respiratory status in a pig model of acute dichlorvos poisoning.</p><p><b>METHODS</b>Twenty female pigs were randomly allocated to dichlorvos (n = 7), atropine (n = 7), and control (n = 6) groups. In the dichlorvos group, pigs were administered 80% emulsifiable dichlorvos (100 mg/kg) via a gastric tube. In the atropine group, pigs were similarly administered dichlorvos, and 0.5 hours later, atropine was injected to attain and maintain atropinization. The control group was administered saline solution. Arterial blood gas was measured at 0, 0.5, 1, 2, 4, and 6 hours post-injection. The extravascular lung water index and pulmonary vascular permeability index were recorded by the pulse-induced contour cardiac output monitor. At termination of the study, the animals were euthanized, the lung wet-to-dry weight ratio was determined, and histopathology was observed.</p><p><b>RESULTS</b>In the dichlorvos group, the extravascular lung water index and pulmonary vascular permeability index were substantially increased from 0.5 hours and were particularly high within 1 hour. In the atropine group, these indices increased initially, but decreased from the 1-hour mark. The control group exhibited no obvious changes. In both the dichlorvos and atropine groups, the extravascular lung water index was negatively correlated with partial pressure of oxygen/fraction of inspiration oxygen (PO2/FiO2) and positively correlated with the pulmonary vascular permeability index. Compared with the control group, the lung wet-to-dry weight ratio markedly increased and the histopathological findings obviously changed in the dichlorvos group, but only mildly increased and changed, respectively, in the atropine group.</p><p><b>CONCLUSION</b>The extravascular lung water index is an appropriate and valuable parameter for assessment of respiratory function in acute dichlorvos poisoning.</p>

Mechanism of apoptosis of NB4 cells induced by arsenic trioxide and cyclooxygenase-2 expression / 中国实验血液学杂志

Objective of this study was to investigate the changes of cyclooxygenase-2 expression and mitochondrial membrane potential in apoptotic NB4 cells induced by arsenic trioxide (As(2)O(3)). The morphological changes in apoptosis process of NB4 cells treated by arsenic trioxide were observed under immunofluorescence microscope and DNA electrophoresis method, and the apoptosis rate of NB4 cells and the variations of mitochondrial membrane potential were detected by flow cytometry. Furthermore, the variations of expression level of cyclooxygenase-2 protein were analyzed by using Western blot method. The results indicated that after NB4 cells were treated with 2 µmol/L As(2)O(3) for 48 hours, some variations of NB4 cells were observed, such as pyknosis, chromatin segmentation, even fragmentation. Meanwhile, the typical DNA Ladder phenomenon was observed. The apoptosis rate of NB4 cells treated with 3 µmol/L As(2)O(3) for 48 hours was 33.34%, Furthermore the apoptosis rate of NB4 cells was enhanced along with the increase of concentration of As(2)O(3). After NB4 cells were treated with 0.5, 1, 2, 4 and 8 µmol/L As(2)O(3) for 48 hours, the mitochondrial membrane potential decreased by 12.8%, 21.6%, 66.9%, 83.7% and 83.8% respectively. The Western blot detection results showed that the expression level of cyclooxygenase-2 protein in NB4 cells was lower than that in control cells and decreased along with the rise of As(2)O(3) concentration, then the negative dose-dependent manner was observed between these 2 groups. It is concluded that As(2)O(3) can effectively induce NB4 cell apoptosis, and the dose-dependent manner existed in certain extent of concentrations. The decrease of mitochondrial membrane potential may be related with NB4 cell apoptosis induced by As(2)O(3). Cyclooxygenase-2 participates in the process of NB4 cell apoptosis induced by As(2)O(3).

Pharmacokinetics of cantide, an antisense oligonucleotide, and its metabolites in rhesus monkeys / 药学学报

To study the pharmacokinetics of cantide, an antisense oligonucleotide, and its metabolites after iv gtt administration in rhesus monkeys, a dual solid phase extraction pretreatment method coupling with non-gel sieving capillary electrophoresis analysis method was used for determination of cantide and its metabolites in plasma and their pharmacokinetic parameters were calculated. The pharmacokinetic behavior of cantide and its metabolites (M1 and M2) after iv gtt administration (8, 16 and 24 mg kg(-1)) in rhesus monkeys were investigated. After iv gtt administration of cantide to rhesus monkeys, cantide in plasma was eliminated rapidly and the terminal elimination half-life (t1/2) was 57.91-77.97 min, the correlation coefficients (r) to the dose of Cmax AUC(o-inf) and AUC(0-t) of the prototype was 0.9918, 0.9568 and 0.9773, respectively. The metabolites of cantide reached the Cmax following cantide immediately and the Cmax of metabolites were lower than that of the prototype. The CL(S) of cantide and its metabolites (M1 and M2) were 1.60-2.19, 5.92-8.58 and 6.07-8.78 mL min(-1) kg(-1), respectively. So, it is concluded that the Cmax of cantide and its metabolites increased with the dose, which is the same as their AUC(0-inf) and AUC(0-t). The CL(S) of metabolites were higher than that of the prototype. The MRT and t1/2 of metabolites in the high dose group increased obviously.

Identification of gene rearrangements in childhood leukemia by using a multiplex polymerase chain reaction-microarray approach / 中国实验血液学杂志

The aim of study was to investigate the application of a novel microarray approach for the eight most common leukemia translocations in children for routine molecular diagnostic hematopathology practice. Bone marrow samples from 84 children with leukemia were analyzed by multiplex nested RT-PCR combined with oligonucleotide microarray. The results showed that out of 84 leukemic samples, 31 (36. 90%) carried 8 types of fusion genes including tel/aml1, e2a/pbx1, bcr/ablp190, bcr/ablp210, mll/af4, aml1/eto, pml/raralpha, cbfbeta/myh11. The sensitivity and specificity of the assay is comparable with the RT-PCR technique. In conclusion, this multiplex nested RT-PCR could quickly screen 8 types of chromosome structural aberrations at the same time. It can provide reliable and helpful information for risk stratification, therapy evaluation and prognosis prediction in childhood leukemia. There are both advantages and disadvantages in applying this new method. The microarray-based assay will be an effective and reliable tool in the clinical screening of leukemia patients for the presence of specific gene rearrangements with important diagnostic and prognostic implications.

Detection of fusion genes in childhood acute lymphoblastic leukemia by using oligonucleotide microarrays approach / 中华儿科杂志

<p><b>OBJECTIVE</b>To evaluate the possibility of applying oligonucleotide microarrays for detection of the fusion genes in childhood acute lymphoblastic leukemia (ALL).</p><p><b>METHODS</b>To detect five types of fusion genes emerging frequently in childhood ALL including TEL/AML1, E2A/PBX1, BCR/ABLp190, BCR/ABLp210, MLL/AF4, probes were designed, synthesized and spotted on the chemical-material-coated-glass plates in array. The total RNAs were extracted from patients' bone marrow or peripheral blood cells at the beginning of diagnosis, analyzed by multiplex nested reverse-transcription-polymerase chain reaction (RT-PCR), and labeled by fluorescein. The products of RT-PCR were hybridized with microarray in order to detect specific types of fusion genes in leukemia cells.</p><p><b>RESULTS</b>Distinctive hybridization signals were obtained for internal positive control and specific types of fusion genes. TEL/AML1 gene was found positive in 2 of the 36 cases, E2A/PBX1 gene in 3, BCR/ABLp190 gene in 2, BCR/ ABLp210 gene in one, and MLL/ AF4 gene in one. The results of the microarray and RT-PCR were consistent.</p><p><b>CONCLUSION</b>The microarray-based assay could screen 5 types of chromosome structural aberrations and the splice variants at the same time. It can provide reliable and helpful information for patient stratification, evaluation of therapeutic effects and prediction of prognosis in childhood ALL, although there are both advantages and disadvantages in applying this new method.</p>

A genetic optimization designing method for microorganism detection genechip probe based on genetic algorithm / 中国医疗器械杂志

A new automatic selection approach of microorganism specific fragment combination is presented in this paper. Genetic algorithm is used to search optimal solution on the basis of classification ability of SNP combination, which is evaluated by the rough set theory. Other related experimental parameters are also been incorporated. Experimental results show that the method can find the best SNP combination pattern efficiently and accurately, which implies that it is a reliable approach to the genechip probe design.

Sequence analysis of the complete genome of bocavirus WLL-1 / 病毒学报

Human bocavirus, which was firstly discovered in 2005, is a new human parvovirus associated with lower respiratory tract infection in children. In this study, a human bocavirus, named WLL-1 isolate, was identified in Wenlin County, Zhejiang Province. The genome of bocavirus WLL-1 has been sequenced and analyzed. Phylogenetic analyses showed that WLL-1 shares 99% homology with other bocaviruses recently reported, but also has some special variations.

Progress in a research on biochip image analysis / 中国医疗器械杂志

Microarray imaging is considered as an important tool for large scale analysis of gene expression. The accuracy of the gene expression depends on the experiment itself and further image processing. It's well known that the image analysis introduced during the experiment will greatly affect the accuracy of the gene expression. Microarray image analysis plays a potentially-arge impact on the gene detecting subsequent analysis (image segmentation, spot intensity and spot intensity extraction). In this paper, based on the characters of microarray images, the methods of microarray images' denoising and automatic-identification of the spot are reviewed.

Establishment and preliminary application of a gene chip for detection of hepatitis B virus "a" determinant hotpoint mutation / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To develop a gene chip for rapid detection of the "a" determinant hotpoint mutation of hepatitis B virus (HBV).</p><p><b>METHODS</b>Primers were designed in the HBV conservative region, and probes for detecting 126A, 126S, 144A, 145R, 145E, 144A+145R, and 144A+145E mutants were developed for that gene chip. PCR amplification and gene chip technology were optimized. The performance of the gene chip was evaluated by detecting the reference plasmids. Forty five samples of serum obtained from patients with chronic hepatitis B were used to compare the sensitivity of the gene chip and the direct sequencing of PCR products.</p><p><b>RESULTS</b>The oligonucleotide microarray was specific for mutant and native plasmids. The sensitivity of the gene chip was 5 x 10(3)copies/micro l with a high reproducibility. The gene chip could detect minor variants when they were more than 10% among the HBV strains. The positive rates of 126A, 126S-1, 126S-2 detected in the 45 specimens by the gene chip (46.67%, 35.56% and 24.44%, respectively) were higher than those detected by direct sequencing of PCR products (9.00%, 4.44% and 2.22%; P=0.000, P=0.000 and P=0.002, respectively). The sequencing of cloned PCR products demonstrated that the gene chip was specific for the "a" determinant hotpoint mutation detection.</p><p><b>CONCLUSION</b>HBV "a" determinant hotpoint mutations can be detected by oligonucleotide microarray with high sensitivity and specificity, providing a method for large scale screening of the mutants.</p>

Preparation and identification of the oligonucleotide chip for detecting the virulence-associated genotypes and drug resistance of Helicobacter pylori / 中国医学科学院学报

<p><b>OBJECTIVE</b>To develop an oligonucleotide microarray-based method for the simultaneous detection of Helicobacter pylori (Hp) infection, virulence-associated genotypes, and drug resistance.</p><p><b>METHODS</b>Hp was classified into cagA + and cagA- genotypes based on its virulence. Clarithromycin-resistance of Hp was identified by existence of point mutations in 23S rRNA. We constructed an oligonucleotide microarray chip to simultaneously diagnose Hp infection and detect its virulence-associated genotypes and mutations associated with clarithromycin-resistance. The diagnostic accuracy of the constructed microarray was tested with templates of wild type and mutated type.</p><p><b>RESULTS</b>The oligonucleotide chip accurately detected cagA + and cagA- genotypes of Hp, as well as four common point mutations in 23S rRNA related to clarithromycin-resistance.</p><p><b>CONCLUSION</b>Oligonucleotide microarray chip can be used to diagnose Hp infection and test its virulence-associated genotypes and drug resistance simultaneously.</p>

Role of mutations on the "hepatitis B virus 'a' determinant hotpoint" to the efficacy of hepatitis B vaccine / 中华流行病学杂志

<p><b>OBJECTIVE</b>To study how hepatitis B virus(HBV) 'a' determinant hotpoint mutations were influecing the hepatitis B vaccine efficacy.</p><p><b>METHODS</b>Primers were designed in HBV conservative region, and the degenerate probes for detecting 16 'a' determinant hotpoint mutations were developed for gene chips. Sensitivity and specificity of the gene chips were evaluated by clone sequencing. Sera of 47 pairs of mothers and infants with immune failure and 323 mothers of children with immune protection of HB vaccine were detected by the gene chips.</p><p><b>RESULTS</b>Result from clone sequencing demonstrated that the gene chips were specific for the detection of 'a' determinant hotpoint mutations. The wild type of HBV was still dominant, with the prevalence of 78.66%, and the mutation frequencies of 126A, 145R, 126S-1, 126S-2, 129H, 144A, and 129R were 11.27%, 5.76%, 5.28%, 4.56%, 1.20%, 0.72% and 0.24%, respectively. The prevalence of 126A mutation was significantly higher than that of other mutations(P < 0.01). No significant differences were found in mother-infant transmission rates of 126A, 126S-1, 126S-2 and 145R variants.</p><p><b>CONCLUSION</b>The currently available hepatitis B vaccine could block mother-infant transmission of 126A, 126S and 145R variants. It appears that there is no need to develop a new hepatitis B vaccine against 126 and 145 variants at present, but the consistent epidemiological surveillance on HBV mutants should be carried out.</p>

Effects of Siwu Tang on protein expression of HFCL / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the effects of Siwu Tang on HFCL proliferation and protein expression of bone marrow stromal cells and to discuss molecular mechanism of enriching blood functions of Siwu Tang.</p><p><b>METHOD</b>MTT, flow cytometry and proteomics were used respectively to measure the effect of Siwu Tang on HFCL proliferation, cell cycle and proteinexpression.</p><p><b>RESULT</b>Siwu Tang can promote HFCL proliferation, increase proliferative index, up-regulate 12 kinds of proteins and down-regulate 5 kinds of proteins of HFCL.</p><p><b>CONCLUSION</b>Through promoting HFCL proliferation and acting on multipleprotein targets, the blood enriching function of Siwu Tang was exerted indirectly.</p>

Effects of fufangdenshen tablets on human atherosclerotic plasma protein using proteomic technique / 中国中药杂志

<p><b>OBJECTIVE</b>To research the effects of fufangdenshen tablets on atherosclerotic plasma protein using proteomic technique and further explore its potential molecular mechanism to cure atherosclerosis.</p><p><b>METHOD</b>The plasma of normal, atherosclerosis and cured group were collected, and the albumin in plasma was removed. Proteomic protocol involved of 2-DE, image analysis and spectrometry detection was used to detect regulated plasma protein by fufangdenshen tablets.</p><p><b>RESULT</b>3 decreased expressed and 6 higher protein in atherosclerotic plasma could be recovered by fufangdenshen tablets. The levels of fibrinogen, Granzyme C and immunoglobulin were decreased by fufangdenshen tablets.</p><p><b>CONCLUSION</b>The levels of fibrinogen, Granzyme C in atherosclerotic plasma could be decreased by fufangdenshen tablets. The effects of fufangdenshen tablets on atherosclerosis included that: inhibition of adhesion of monocyte, inhibition of proliferation and migration of VSMC, and weakening of inflammation.</p>

Screening and their anti-tumor activity of antisense oligodeoxynucleotides targeting KDR mRNA in breast cancer MCF-7 cells / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To screen the antisense oligodeoxynucleotides (asONs) which could hybridize with KDR (kinase insert domain-containing receptor) mRNA in an effective and specific way and to explore their anti-tumor effects on breast cancer MCF-7 cell line in vitro.</p><p><b>METHODS</b>The asONs were firstly selected using oligodeoxynucleotides library hybridization or computer prediction, then their hybridization ability with KDR mRNA was further tested with oligonucleotide microarray. The asONs with strong hybridization intensity were selected. Their inhibitory effects on MCF-7 cells proliferation and KDR expression were assayed by MTT, RT-PCR and Western blotting assay, respectively.</p><p><b>RESULTS</b>In 13 asONs selected with oligodeoxynucleotides library hybridization, 8 (8/13, 61.5%) showed strong hybridization signals, while such was only 1 in 17 asONs designed by computer prediction. 9 asONs with strong hybridization intensity were selected and synthesized with phosphorothioated modification. All these asONs inhibited the MCF-7 cells proliferation in a dose-dependent manner, in which asON4 and asON7 screened by oligodeoxynucleotides library in combination with oligonucleotide microarray were the most effective, with inhibitory rates of 51.6% and 62.2% at 0.8 micromol/L, respectively. The KDR expression at mRNA and protein levels was reduced by both the two asONs, in a dose-dependent manner.</p><p><b>CONCLUSION</b>asONs screened by oligodeoxynucleotides library hybridization are well consistent with that chosen with oligonucleotide microarray. The combination of oligodeoxynucleotides library with oligonucleotide microarray is an effective approach of asONs screening. The asONs targeting KDR mRNA showed prominent anti-tumor activity on breast cancer MCF-7 cells.</p>

Influence of cytochrom P450 CYP2C9 polymorphism on the pharmacokinetics of tolbutamide metabolism using oligonucleotide genotyping microarray / 药学学报

<p><b>AIM</b>To investigate the influence of cytochrom P450 CYP2C9 polymorphism on the pharmacokinetics of tolbutamide.</p><p><b>METHODS</b>An oligonucleotide microarray was designed and fabricated to genotype the CYP2C9 accurately and quickly. 137 healthy volunteers were genotyped with the array to investigate the frequency of CYP2C9 functional SNPs. Moreover, 1 homozygous mutant, 9 heterozygous and 10 wild-genotypes subjects in the assay were selected randomly and sequenced directly. After orally taking tolbutamide, blood samples and urine samples were collected, and their pharmacokinetics was studied with HPLC.</p><p><b>RESULTS</b>CYP2C9 *1/*3 were found in 9 of 137 volunteers, CYP2C9 *3/*3 in only one, others were all CYP2C9 *1/*1 wild types. CYP2C9 *2, CYP2C9 *4 and CYP2C9 *5 alleles were not detected. Direct sequencing of the purified PCR products of the heterozygotes, mutant homozygotes and ten wild type individuals gave a corresponding result to that genotyped by microarray. Pharmacokinetic outcome showed that the individuals with CYP2C9 *1/*3 or CYP2C9 *3/*3 had slower metabolic elimination of tolbutamide than those with CYP2C9 *1/*1.</p><p><b>CONCLUSION</b>CYP2C9 genetic polymorphism has a significant influence on the pharmacokinetics of tolbutamide. Pharmacogenomic study will be helpful in guiding rational and individualized medication. Key words: tolbutamide; cytochrom P450 CYP2C9; allele; single nucleotide polymorphism; genotyping</p>

The effect of Siwu Tang on EPO and G-CSF gene expression in bone marrow of irradiated blood deficiency mice / 中国中药杂志

<p><b>OBJECTIVE</b>To observe the blood enriching function of Siwu Tang and its effect on Epo and G-CSF gene expressions in bone marrow of blood deficiency mice, and thus provide the basis for understanding the molecular mechanism of blood enriching function of Siwu Tang.</p><p><b>METHOD</b>The animal model of blood deficiency were established in the mice by using 3.5 Gy60Co gamma-ray. Peripheral blood cells were analyzed and CFU-GM, BFU-E, CFU-E and CFU-mix were counted in bone marrow colony cultured. Both Epo and G-CSF gene expressions in bone marrow were measured with RT-PCR.</p><p><b>RESULT</b>Siwu Tang significantly increased the number of peripheral blood cells and the amount of CFU-GM, BFU-E, CFU-E and CFU-mix in bone marrow and enhanced Epo and G-CSF gene expression in bone marrow in the mice with blood deficiency.</p><p><b>CONCLUSION</b>The promotion of Epo and G-CSF gene expressions in bone marrow may be one of the mechanisms underlying the blood enriching function of Siwu Tang decoction.</p>

Identification of acute leukemia-specific genes from leukemia recipient/sibling donor pairs by distinguishing study with oligonucleotide microarrays / 中国实验血液学杂志

To explore differentially expressed genes in leukemia gene expression profile and identify main related genes in acute leukemia, gene expression profiles were analyzed in bone marrow/leucopheresis peripheral blood stem cells samples from 9 acute leukemia patients and their sibling donors with the use of oligonucleotide microarrays. 163 reported leukemia-related genes were involved in the study. The oligonucleotide primers were designed, synthesized and spotted on the chemical-material-coated-glass plates in array. The total RNAs were isolated from nine patients' bone marrow or leucopheresis peripheral blood cells and from nine their sibling donors peripheral blood stem cells treated by G-CSF, then collected by CS-3000 cell selection machine, and were reversely transcribed to cDNAs with the incorporations of fluorescent dUTP. The mixed probes were then hybridized to the oligonucleotide microarray. The results showed that in four patient/donor pairs with B-ALL, 5 up-regulated (RIZ, STK-1, T-cell leukemia/lymphoma 1A, Cbp/p300, Op18) and 1 down-regulated genes (hematopoietic proteoglycan core protein) were identified; In five patient/donor pairs with AML-M(4) and AML-M(5), 6 up-regulated (STAT5B, ligand p62 for the Lck SH2, CST3, LTC4S, myeloid leukemia factor 2 and epb72) and 1 down-regulated genes (CCR5) were identified. In conclusion, on the basis of distinguishing study of specific genetic related recipient/sibling donor pairs, screening leukemia-related genes with oligonucleotide microarrays, a set of 13 up-regulated or down-regulated genes among 163 leukemia-related genes has been identified. The result has further confirmed that above genes play critical role in the molecular mechanism of acute leukemia.

Identification bcr-abl fusion gene in leukemia cells with oligonucleotide microarray / 中国实验血液学杂志

To explore the application value of bcr-abl fusion gene deterction microarray in diagnosis, typing, choosing of treatment variant and prognosis judgment, probe for fusion gene detection was designed, oligonucleotide microarray was prepared; total RNA was extracted, reverse-transcripted and labeled by fluorescence, then cDNA was hybridized with microarray in order to detect bcr-abl fusion gene in leukemia cells. The results showed that better reaction conditions were gained by exploration of hybridizotion temperature and elution conditions, bcr-abl fusion gene in leukemia cells was detected by prepared miccroarray. In conclusion, oligonucleotide microarray is effective in detecting the fusion gene and has some unique advantages and certain clinical application value, but has some deficiency too. If microarray can be improved further, it could be used widely in the field of hematology.