Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 20 de 45
Filter
1.
Front Bioeng Biotechnol ; 11: 1310247, 2023.
Article in English | MEDLINE | ID: mdl-38239918

ABSTRACT

Introduction: Small-scaled robotic walkers play an increasingly important role in Activity of Daily Living (ADL) assistance in the face of ever-increasing rehab requirements and existing equipment drawbacks. This paper proposes a Rehabilitation Robotic Walker (RRW) for walking assistance and body weight support (BWS) during gait rehabilitation. Methods: The walker provides the patients with weight offloading and guiding force to mimic a series of the physiotherapist's (PT's) movements, and creates a natural, comfortable, and safe environment. This system consists of an omnidirectional mobile platform, a BWS mechanism, and a pelvic brace to smooth the motions of the pelvis. To recognize the human intentions, four force sensors, two joysticks, and one depth-sensing camera were used to monitor the human-machine information, and a multimodal fusion algorithm for intention recognition was proposed to improve the accuracy. Then the system obtained the heading angle E, the pelvic pose F, and the motion vector H via the camera, the force sensors, and the joysticks respectively, classified the intentions with feature extraction and information fusion, and finally outputted the motor speed control through the robot's kinematics. Results: To validate the validity of the algorithm above, a preliminary test with three volunteers was conducted to study the motion control. The results showed that the average error of the integral square error (ISE) was 2.90 and the minimum error was 1.96. Discussion: The results demonstrated the efficiency of the proposed method, and that the system is capable of providing walking assistance.

2.
Mol Immunol ; 117: 20-28, 2020 01.
Article in English | MEDLINE | ID: mdl-31731055

ABSTRACT

BACKGROUND: LncRNA NEAT1 was associated with the tumorigenesis of multiple myeloma (MM). However, the mechanisms of M2 macrophage polarization involved with NEAT1 in MM are still unknown. METHODS: Bone marrow samples, multiple myeloma cells RPMI 8226 and monocyte cell line THP-1 were used in this study. The expression of NEAT1 and miR-214 was modified by transfection with the shNEAT1 or miR-214 inhibitor. The expression of NEAT1, miR-214 and B7-H3 in MM patient tissues and cells was analyzed by RT-qPCR. ELISA assay was used to determine the release of B7-H3 in the supernatant of cell culture. The patient survival curve was analyzed using Kaplan-Meier method. The macrophage polarization markers were examined by RT-qPCR and western blotting. The interaction between NEAT1, miR-214 and B7-H3 was analyzed by Dual-Luciferase reporter and RIP assays. AG490 was used to block the JAK2/STAT3 signaling. Co-culture of THP-1 and RPMI 8226 cells was used for macrophage polarization. RESULTS: NEAT1 and B7-H3 were up-regulated, but miR-214 was obviously down-regulated in MM patients. B7-H3, NEAT1 and miR-214 were associated with overall survival time of MM patients. NEAT1 silencing induced miR-214 and inhibited the expression and release of B7-H3 and then suppressed M2 macrophage polarization via inhibiting the JAK2/STAT3 signaling. NEAT1 directly targeted miR-214, and miR-214 directly bound to B7-H3. MiR-214 inhibitor reversed the down-regulation and release of B7-H3 and M2 macrophage polarization caused by shNEAT1. The specific JAK2/STAT3 signaling inhibitor AG490 abrogated M2 macrophage polarization. CONCLUSION: NEAT1 promoted M2 macrophage polarization by sponging miR-214 and then regulating B7-H3, thus accelerating MM progression via the JAK2/STAT3 signaling pathway. Our study revealed novel mechanisms of M2 macrophage polarization and provided new potential clinical therapeutic targets for MM.


Subject(s)
B7 Antigens/immunology , Macrophages/immunology , MicroRNAs/immunology , Multiple Myeloma/immunology , RNA, Long Noncoding/immunology , B7 Antigens/metabolism , Cell Differentiation/immunology , Cell Line, Tumor , Gene Expression Regulation, Neoplastic/immunology , Humans , Macrophage Activation , Macrophages/metabolism , MicroRNAs/metabolism , Multiple Myeloma/metabolism , RNA, Long Noncoding/metabolism
3.
BMC Genomics ; 19(1): 411, 2018 May 29.
Article in English | MEDLINE | ID: mdl-29843600

ABSTRACT

BACKGROUND: Chloroplasts have their own genomes, independent from nuclear genomes, that play vital roles in growth, which is a major targeted trait for genetic improvement in Populus. Angiosperm chloroplast genomes are maternally inherited, but the chloroplast' variation pattern of poplar at the single-base level during the transmission from mother to offspring remains unknown. RESULTS: Here, we constructed high-quality and almost complete chloroplast genomes for three poplar clones, 'NL895' and its parents, 'I69' and 'I45', from the short-read datasets using multi-pass sequencing (15-16 times per clone) and ultra-high coverage (at least 8500× per clone), with the four-step strategy of Simulation-Assembly-Merging-Correction. Each of the three resulting chloroplast assemblies contained contigs covering > 99% of Populus trichocarpa chloroplast DNA as a reference. A total of 401 variant loci were identified by a hybrid strategy of genome comparison-based and mapping-based single nucleotide polymorphism calling. The genotypes of 94 variant loci were different among the three poplar clones. However, only 1 of the 94 loci was a missense mutation, which was located in the exon region of rpoC1 encoding the ß' subunit of plastid-encoded RNA polymerase. The genotype of the loci in NL895 and its female parent (I69) was different from that of its male parent (I45). CONCLUSIONS: This research provides resources for further chloroplast genomic studies of a F1 full-sibling family derived from a cross between I69 and I45, and will improve the application of chloroplast genomic information in modern Populus breeding programs.


Subject(s)
Genome, Chloroplast/genetics , Mutation , Populus/genetics , DNA, Chloroplast/genetics , Genome Size , Genotype , Polymorphism, Single Nucleotide
4.
Ann Dermatol ; 30(2): 129-135, 2018 Apr.
Article in English | MEDLINE | ID: mdl-29606808

ABSTRACT

Tumor suppressor gene O6-methylguanine-DNA methyltransferase (MGMT) promoter methylation has been reported in melanoma. However, the clinical and prognostic significance of MGMT promoter methylation in patients with melanoma remained to be determined. A systematic search was performed to identify eligible papers published. The overall odds ratios (ORs) or hazard ratios and their 95% confidence intervals were calculated. Final 12 eligible publications involving Caucasian population were performed in this study, including 1,071 metastatic melanoma patients, 154 primary melanoma patients, and 211 normal controls. MGMT promoter methylation was significantly higher in primary or metastatic melanoma than in normal controls (p<0.05). No difference of MGMT promoter methylation was found in primary and metastatic melanoma (p=0.432). When metastatic melanoma was compared to normal controls, subgroup analysis showed the correlation between MGMT promoter methylation and different sample materials (tissue: OR=7.01, p<0.001 and blood: OR=12.04, p=0.005). MGMT promoter methylation was not associated with response to drug therapy and the prognosis in overall survival and progression-free survival for multivariate analysis. Our results show that MGMT promoter methylation may be correlated with the increased risk of primary or metastatic melanoma. Based on blood samples, MGMT promoter methylation may become a noninvasive biomarker for the detection of metastatic melanoma. Further additional clinical studies are necessary.

5.
Cancer Invest ; 36(2): 141-151, 2018 Feb 07.
Article in English | MEDLINE | ID: mdl-29420094

ABSTRACT

This study systematically reviewed previous literatures and analyzed the genotype-phenotype relationship between the multiple endocrine neoplasia type 2A (MEN 2A)-cutaneous lichen amyloidosis (CLA) and RET/OSMR/IL31RA mutations. RET/OSMR/IL31RA screening was performed on 8 RET-carriers from 3 independent Chinese MEN 2A families. Besides, 51 MEN 2A-CLA patients in 116 RET carriers from literatures were clustered and analyzed. Our results indicated that almost all MEN 2A-CLA patients exhibited CLA which was located in the scapular region and carried RET mutation at codon 634. Meanwhile, we firstly described MEN 2A-CLA here in Chinese Han patient with RET p.C634F mutation.


Subject(s)
Amyloidosis/complications , Asian People/genetics , Genetic Markers , Multiple Endocrine Neoplasia Type 2a/complications , Mutation , Proto-Oncogene Proteins c-ret/genetics , Skin Diseases, Metabolic/complications , Adult , Amyloidosis/genetics , Child , Female , Humans , Male , Middle Aged , Multiple Endocrine Neoplasia Type 2a/genetics , Oncostatin M Receptor beta Subunit/genetics , Pedigree , Phenotype , Proto-Oncogene Mas , Receptors, Interleukin/genetics , Skin Diseases, Metabolic/genetics
6.
PLoS One ; 12(3): e0172475, 2017.
Article in English | MEDLINE | ID: mdl-28323890

ABSTRACT

Liriodendron is a genus of the magnolia family comprised of two flowering tree species that produce hardwoods of great ecological and economic value. However, only a limited amount of genetic research has been performed on the Liriodendron genus partly because transient or stable transgenic trees have been difficult to produce. In general, transient expression systems are indispensable for rapid, high-throughput screening and systematic characterization of gene functions at a low cost; therefore, development of such a system for Liriodendron would provide a necessary step forward for research on Magnoliaceae and other woody trees. Herein, we describe an efficient and rapid protocol for preparing protoplasts from the leaf mesophyll tissue of a Liriodendron hybrid and an optimized system for polyethylene glycol-mediated transient transfection of the protoplasts. Because the leaves of the Liriodendron hybrid are waxy, we formulated an enzyme mix containing 1.5% (w/v) Cellulase R-10, 0.5% (w/v) Macerozyme R-10, and 0.1% (w/v) Pectolyase Y-23 to efficiently isolate protoplasts from the Liriodendron hybrid leaf mesophyll tissue in 3 h. We optimized Liriodendron protoplast transfection efficiency by including 20 µg plasmid DNA per 104 protoplasts, a transformation time of 20 min, and inclusion of 20% (w/v) polyethylene glycol 4000. After integrating the Liriodendron WOX1 gene into pJIT166-GFP to produce a WOX1-GFP fusion product and transfecting it into isolated protoplasts, LhWOX1-GFP was found to localize to the nucleus according to its green fluorescence.


Subject(s)
Gene Expression , Liriodendron , Mesophyll Cells , Protoplasts , Transfection , Cell Culture Techniques , Genetic Vectors , Luminescent Proteins/genetics , Luminescent Proteins/metabolism , Mesophyll Cells/metabolism , Plant Leaves , Plant Proteins/genetics , Plant Proteins/metabolism , Protoplasts/metabolism
7.
Australas J Dermatol ; 58(1): e5-e7, 2017 Feb.
Article in English | MEDLINE | ID: mdl-26507226

ABSTRACT

We describe two rarely documented cases of telangiectasia macularis multiplex acquisita (TMMA) with a history of hepatitis B infection. Both patients presented with multiple erythematous macules and telangiectasia on bilateral upper arms and the upper part of the trunk. Patient 1 also had spider naevi on the upper part of the chest and palmar erythema; thus we inferred that TMMA, like spider naevi and palmar erythema, might belong to the spectrum of vascular changes of liver diseases.


Subject(s)
Erythema/complications , Hepatitis B, Chronic/complications , Telangiectasis/complications , Erythema/pathology , Humans , Male , Middle Aged , Telangiectasis/pathology
8.
Front Plant Sci ; 7: 826, 2016.
Article in English | MEDLINE | ID: mdl-27379122

ABSTRACT

Nectar is a primary nutrient reward for a variety of pollinators. Recent studies have demonstrated that nectar also has defensive functions against microbial invasion. In this study, the Liriodendron tulipifera nectary was first examined by scanning electron microscopy, and then the nectar was analyzed by two-dimensional gel electrophoresis and liquid chromatography-tandem mass spectrometry, which led to identification of 42 nectar proteins involved in various biological functions. Bioinformatic analysis was then performed on an identified novel rubber elongation factor (REF) protein in L. tulipifera nectar. The protein was particularly abundant, representing ∼60% of the major bands of 31 to 43 kDa, and showed high, stage-specific expression in nectary tissue. The REF family proteins are the major allergens in latex. We propose that REF in L. tulipifera nectar has defensive characteristics against microorganisms.

9.
Shanghai Kou Qiang Yi Xue ; 25(1): 91-6, 2016 Feb.
Article in Chinese | MEDLINE | ID: mdl-27063317

ABSTRACT

PURPOSE: To investigate the effects of different psychological interventions on relieving orthodontic pain in patients with different personalities. METHODS: Three hundred patients were involved and randomized into five groups:control group, cognitive therapy group, music therapy group, muscle relaxation group and suggestion therapy group. Eysenck personality questionnaire was used to evaluate personality traits of patients, and visual analogue scale (VAS) was used to assess patients' intensity of orthodontic pain at 2, 6, 12, 24, 48 and 72 h after initial archwire placement.The VAS scores were analyzed via repeated measures analysis of variance with SPSS 16.0 software package. RESULTS: The cognitive group,music group, muscle relaxation group and suggestion group were reported lower pain than control group (P<0.001). The music group showed a greater decrease in VAS than other four groups in patients with a tendency of extroversion and stable mood (P<0.01).The suggestion group showed less pain than cognitive group in patients with a tendency of extroversion and unstable mood (P<0.05).For patients with other personality traits, there was no significant difference among the four intervention groups. CONCLUSIONS: Cognitive therapy, music therapy, muscle relaxation and suggestion therapy could relieve orthodontic pain effectively. For patients with a tendency of extroversion and stable mood, music therapy was the first choice to control orthodontic pain. Cognitive therapy could be used for patients with other personality traits.


Subject(s)
Analgesia/methods , Pain Measurement/methods , Pain/psychology , Analgesics , Humans , Music Therapy , Pain Management , Personality
11.
Biochem Biophys Res Commun ; 472(4): 662-8, 2016 Apr 15.
Article in English | MEDLINE | ID: mdl-26920060

ABSTRACT

Akt signaling plays a pivotal role in acute myeloid leukemia (AML) development and progression. In the present study, we evaluated the potential anti-AML activity by a novel Akt kinase inhibitor A-674563. Our results showed that A-674563 dose-dependently inhibited survival and proliferation of U937 AML cells and six lines of human AML progenitor cells, yet sparing human peripheral blood mononuclear leukocytes (PBMCs). A-674563 activated caspase-3/9 and apoptosis in the AML cells. Reversely, the pan-caspase inhibitor z-VAD-CHO dramatically alleviated A-674563-induced AML cell apoptosis and cytotoxicity. For the molecular study, we showed that A-674563 blocked Akt activation in U937 cells and human AML progenitor cells. Further, A-674563 decreased sphingosine kinase 1 (SphK1) activity in above AML cells to deplete pro-survival sphingosine-1-phosphate (S1P) and boost pro-apoptotic ceramide production. Such an effect on SphK1 signaling by A-674563 appeared independent of Akt blockage. Significantly, K6PC-5, a novel SphK1 activator, or supplement with S1P attenuated A-674563-induced ceramide production, and subsequent U937 cell death and apoptosis. Importantly, intraperitoneal injection of A-674563 at well-tolerated doses suppressed U937 leukemic xenograft tumor growth in nude mice, whiling significantly improving the animal survival. The results of the current study demonstrate that A-674563 exerts potent anti-leukemic activity in vitro and in vivo, possibly via concurrent targeting Akt and SphK1 signalings.


Subject(s)
Antineoplastic Agents/therapeutic use , Indazoles/therapeutic use , Leukemia, Myeloid, Acute/drug therapy , Phosphotransferases (Alcohol Group Acceptor)/antagonists & inhibitors , Protein Kinase Inhibitors/therapeutic use , Proto-Oncogene Proteins c-akt/antagonists & inhibitors , Pyridines/therapeutic use , Animals , Antineoplastic Agents/pharmacology , Apoptosis/drug effects , Caspase 3/metabolism , Cell Proliferation/drug effects , Cells, Cultured , Humans , Indazoles/pharmacology , Leukemia, Myeloid, Acute/metabolism , Leukemia, Myeloid, Acute/pathology , Male , Mice , Mice, Nude , Phosphotransferases (Alcohol Group Acceptor)/metabolism , Protein Kinase Inhibitors/pharmacology , Proto-Oncogene Proteins c-akt/metabolism , Pyridines/pharmacology , U937 Cells
12.
Ther Innov Regul Sci ; 50(1): 74-81, 2016 Jan.
Article in English | MEDLINE | ID: mdl-30236012

ABSTRACT

BACKGROUND: The 12th Five-Year Plan period is a critical period for the development of the pharmaceutical industry in China. A major focus coming with the New Healthcare Reform is a product structure adjustment in the pharmaceutical industry. Many researchers have been attracted in recent years to product structure adjustment studies. METHODS: An empirical analysis of resources of the New Healthcare Reform was conducted by employing clustering analysis. A panel data model was established, with independent variables consisting of the city residents' medical insurance fund, drug fees per capita, city medical insurance fees per capita, rural medical insurance fees per capita, and number of people benefiting from the new rural cooperative medical scheme and the dependent variable being China's medical technology advancement fees. This study covered 29 provinces and regions and used panel data from 2007 to June 2012 to establish a fundamental regression equation for quantitative analysis. RESULTS: The city residents' medical insurance fund, rural medical insurance fees, and the new rural cooperative medical scheme are significant factors for pharmaceutical structure adjustment in China. The rural medical services market is the most important target market for pharmaceutical industrial transformation. CONCLUSION: The fundamental drug system and medical insurance system, which the New Healthcare Reform focuses upon, are closely related to the pharmaceutical product structure. For entities in which structural adjustment is not autonomous, changes can be achieved through sound coordination between organizational structure, technical structure, and repeated evaluation of consistency of drug product quality and overseas technical license.

13.
Planta ; 242(6): 1361-90, 2015 Dec.
Article in English | MEDLINE | ID: mdl-26232921

ABSTRACT

MAIN CONCLUSION: NO acts as the essential signal to enhance poplar tolerance to chilling stress via antioxidant enzyme activities and protein S -nitrosylation modification, NO signal is also strictly controlled by S -nitrosoglutathione reductase and nitrate reductase to avoid the over-accumulation of reactive nitrogen species. Poplar (Populus trichocarpa) are fast growing woody plants with both ecological and economic value; however, the mechanisms by which poplar adapts to environmental stress are poorly understood. In this study, we used isobaric tags for relative and absolute quantification proteomic approach to characterize the response of poplar exposed to cold stress. We identified 114 proteins that were differentially expressed in plants exposed to cold stress. In particular, some of the proteins are involved in reactive oxygen species (ROS) and reactive nitrogen species (RNS) metabolism. Further physiological analysis showed that nitric oxide (NO) signaling activated a series of downstream defense responses. We further demonstrated that NO activated antioxidant enzyme activities and S-nitrosoglutathione reductase (GSNOR) activities, which would reduce ROS and RNS toxicity and thereby enhance poplar tolerance to cold stress. Suppressing NO accumulation or GSNOR activity aggravated cold damage to poplar leaves. Moreover, our results showed that RNS can suppress the activities of GSNOR and NO nitrate reductase (NR) by S-nitrosylation to fine-tune the NO signal and modulate ROS levels by modulating the S-nitrosylation of ascorbate peroxidase protein. Hence, our data demonstrate that NO signaling activates multiple pathways that enhance poplar tolerances to cold stress, and that NO signaling is strictly controlled through protein post-translational modification by S-nitrosylation.


Subject(s)
Aldehyde Oxidoreductases/metabolism , Cold Temperature , Nitric Oxide/physiology , Populus/physiology , Proteomics , Signal Transduction/physiology , Stress, Physiological/physiology , Populus/enzymology , Reactive Nitrogen Species/metabolism
15.
PLoS One ; 9(2): e89289, 2014.
Article in English | MEDLINE | ID: mdl-24586663

ABSTRACT

PIN-FORMED 1 (PIN1) is an important secondary transporter and determines the direction of intercellular auxin flow. As PIN1 performs the conserved function of auxin transport, it is expected that the sequence and structure of PIN1 is conserved. Therefore, we hypothesized that PIN1 evolve under pervasive purifying selection in the protein-coding sequences in angiosperm. To test this hypothesis, we performed detailed evolutionary analyses of 67 PIN1 sequences from 35 angiosperm species. We found that the PIN1 sequences are highly conserved within their transmembrane regions, part of their hydrophilic regions. We also found that there are two or more PIN1 copies in some of these angiosperm species. PIN1 sequences from Poaceae and Brassicaceae are representative of the modern clade. We identified 12 highly conserved motifs and a significant number of family-specific sites within these motifs. One family-specific site within Motif 11 shows a different residue between monocots and dicots, and is functionally critical for the polarity of PIN1. Likewise, the function of PIN1 appears to be different between monocots and dicots since the phenotype associated with PIN1 overexpression is opposite between Arabidopsis and rice. The evolution of angiosperm PIN1 protein-coding sequences appears to have been primarily driven by purifying selection, but traces of positive selection associated with sequences from certain families also seem to be present. We verified this observation by calculating the numbers of non-synonymous and synonymous changes on each branch of a phylogenetic tree. Our results indicate that the evolution of angiosperm PIN1 sequences involve strong purifying selection. In addition, our results suggest that the conserved sequences of PIN1 derive from a combination of the family-specific site variations and conserved motifs during their unique evolutionary processes, which is critical for the functional integrity and stability of these auxin transporters, especially in new species. Finally, functional difference of PIN1 is likely to be present in angiosperm because the positive selection is occurred in one branch of Poaceae.


Subject(s)
Evolution, Molecular , Magnoliopsida/metabolism , Phylogeny , Plant Proteins/genetics , Brassicaceae/genetics , Magnoliopsida/genetics , Plant Proteins/classification , Poaceae/genetics
16.
Zhong Nan Da Xue Xue Bao Yi Xue Ban ; 37(8): 771-6, 2012 Aug.
Article in Chinese | MEDLINE | ID: mdl-22954922

ABSTRACT

OBJECTIVE: To screen aptamers binding CD33+/CD34- cells from patients with acute myeloblastic leukemia M2 subtype (AML-M2). METHODS: CD33+/CD34- cells from patients with AML-M2 were taken as targeted cells, CD33+/ CD34- cells from normal people were taken as anti-selecting cells, and aptamers in the single strand deoxyribonucleic acid (ssDNA) library were then selected repeatedly by cell-systematic evolution of ligands by exponential enrichment (C-SELEX) technology, and amplified by polymerase chain reaction (PCR) to generate sub-ssDNA library. During the experiment, PCR amplification with fluorescently labeled primer and flow cytometry were performed to analyze the aptamers'enrichment of sub-library, and the final round product of the sub-ssDNA library was cloned. After the sequencing, the primary and secondary structures of the aptamers were analyzed. RESULTS: Electrophoresis indicated that the product of PCR amplification for each round subssDNA library was able to see a clear DNA band in the agarose gel. After 13 rounds of screening, the fluorescence intensity of the sub-ssDNA library binding the cells ranged from 2.14% to 51.12%, reaching a steady state at the 13th round. A total of 30 clones were selected and sequenced, 22 of which contained 1 of the 4 conserved sequences of AAGTA, TATCT, AGATG and AAATT in their primary structure, but the remained eight aptamers contained none of the conserved sequence. Secondary structure analysis indicated that four stem-loops and loop simulation convex structures existed in the aptamers. CONCLUSION: C-SELEX technology can be used to screen the aptamers binding primary cells from patients with leukemia. The aptamers selected from the CD33+/CD34- cells from the patients of AML-M2 subtype might be used for the diagnosis and treatment for leukemia.


Subject(s)
Antigens, Differentiation, Myelomonocytic/genetics , Aptamers, Nucleotide/metabolism , DNA, Single-Stranded/genetics , Leukemia, Myeloid, Acute/genetics , Adolescent , Adult , Antigens, CD34/genetics , Antigens, CD34/immunology , Antigens, Differentiation, Myelomonocytic/immunology , Aptamers, Nucleotide/genetics , Female , Humans , Leukemia, Myeloid, Acute/immunology , Male , Middle Aged , SELEX Aptamer Technique , Sialic Acid Binding Ig-like Lectin 3/genetics , Sialic Acid Binding Ig-like Lectin 3/immunology , Young Adult
17.
Zhonghua Xue Ye Xue Za Zhi ; 33(7): 541-5, 2012 Jul.
Article in Chinese | MEDLINE | ID: mdl-22967414

ABSTRACT

OBJECTIVE: To screen and analyze CD34(+) cell specific microRNAs (miRNAs) from the patients with acute myelogenous leukemia (AML) and their expression. METHODS: CD34(+) cells were sorted from AML patients or the mobilized peripheral blood of the donors of hematopoietic stem cell transplantation (normal control subjects) and followed by the extraction of the cell total RNAs. The differentially expressed microRNAs (miRNAs, miR) were selected after hybridizing with miRNA microarray, real time polymerase chain reaction (real-time PCR) was subsequently applied to confirm the expression of the selected miRs, and PCR products were further cloned and sequenced to check their specificity. RESULTS: Of the differentially expressed miRNAs, 191 were found to be at least one-fold change in the CD34(+) cells between the AML patients and the normal control subjects. Of the 191 miRNAs, the expression difference of 94 was significant (P < 0.05). Among these 94 miRNAs, the expression of 44 miRNAs was increased and the other 50 miRNAs was decreased in the CD34(+) cells from the bone marrow of AML patients compared with the CD34(+) cells from the mobilized peripheral blood of the normal control subjects. Real time PCR verified that the expression level of miR-10a and miR-220c in the CD34(+) cells from the bone marrow of AML patients was 19.6% and 19.0% of that of CD34(+) cells from mobilized peripheral blood of the normal control subjects. DNA sequencing and BLAST DNA database searching results indicated that the PCR products were really miR-10a and miR-220c. CONCLUSION: A variety of differentially expressed-miRNAs are existed between AML and normal control subjects CD34(+) cells, the expression of miR-10a and miR-220c was significantly down-regulated in the CD34(+) cells from the bone marrow of AML patients.


Subject(s)
Hematopoietic Stem Cells , Leukemia, Myeloid, Acute/genetics , MicroRNAs/metabolism , Antigens, CD34/metabolism , Female , Humans , Leukemia, Myeloid, Acute/metabolism , Male , MicroRNAs/genetics , Middle Aged , Oligonucleotide Array Sequence Analysis
18.
Zhongguo Shi Yan Xue Ye Xue Za Zhi ; 19(4): 921-5, 2011 Aug.
Article in Chinese | MEDLINE | ID: mdl-21867615

ABSTRACT

This study was aimed to investigate the expressions of livin and survivin in adult patients with acute lymphoblastic leukemia (ALL) and clinical significance. The expressions of livin and survivin mRNA in bone marrow mononuclear cells of 95 adult ALL patients including 52 de novo patients, 23 relapsed patients and 20 patients with complete remission (CR), and 20 healthy adults as normal controls were detected by using RT-PCR. The results indicated that the expression of livin and survivin mRNA in de novo and relapsed patients was higher than that in normal controls and patients with CR. There were no significant relation of expression level with clinical features such as age, sex, type and de novo leukocyte level. The CR rate in de novo adult ALL patients with positive expression of livin and survivin was lower than that in adult ALL patients with negative gene expression. No relation of mRNA expression between livin and survivin was found in de novo adult ALL patients. It is concluded that genes livin and survivin may be involved in the pathogenesis and progression of adult ALL. Overexpression of livin or survivin may show poor progression. There is no relation of expression between genes livin and survivin in de novo adult ALL patients.


Subject(s)
Adaptor Proteins, Signal Transducing/metabolism , Inhibitor of Apoptosis Proteins/metabolism , Neoplasm Proteins/metabolism , Precursor Cell Lymphoblastic Leukemia-Lymphoma/genetics , Adaptor Proteins, Signal Transducing/genetics , Adolescent , Adult , Aged , Case-Control Studies , Female , Gene Expression , Humans , Inhibitor of Apoptosis Proteins/genetics , Male , Middle Aged , Neoplasm Proteins/genetics , Prognosis , RNA, Messenger/genetics , Reverse Transcriptase Polymerase Chain Reaction , Survivin , Young Adult
19.
Zhongguo Shi Yan Xue Ye Xue Za Zhi ; 19(3): 561-5, 2011 Jun.
Article in Chinese | MEDLINE | ID: mdl-21729522

ABSTRACT

A little is known about the specific marker on the surface of acute leukemia cells, leading to the lack of the specific diagnosis method for acute leukemia. Therefore, in this study, cell-systematic evolution of ligands by exponential enrichment (cSELEX) was performed to screen the aptamers binding to CD33(+)/CD34(+) cells from the patients with acute myeloblastic leukemia (AML) of M(2) subtype (AML-M2) so as to provide the basis for finding the specific marker on the surface of AML-M(2) CD33(+)/CD34(+) cells. Firstly, AML-M2 CD33(+)/CD34(+) cells were sorted and used as targeted cells, and normal CD33(+)/CD34(+)cells were used as counter-targeted cells; the aptamers binding to CD33(+)/CD34(+) cells from patients with AML-M2 were screened from the single strand deoxyribonucleic acid (ssDNA) library by cSELEX. Subsequently, each aptamer structure was analyzed after cloning and sequencing. The results indicated that after 13 round of screenings, the enrichment of aptamers in the ssDNA library was ranged from 0.7% to 52.9%, and reached steady state at 13th round screening. Sequence analysis for 30 aptamers showed that most of the aptamers born one of the three conserved sequences of CCCCT, CTCTC, and CTCAC. Secondary structure analysis indicated that three different secondary structures existed in these aptamers. It is concluded that the aptamers binding to the AML-M(2) CD33(+)/CD34(+) cells are successfully screened, which lay the basis for further looking for the specific marker on the surface of AML-M2 CD33(+)/CD34(+) cells, and the molecular diagnosis of the AML-M2 leukemia.


Subject(s)
Antigens, CD34/genetics , Antigens, CD/genetics , Antigens, Differentiation, Myelomonocytic/genetics , Aptamers, Nucleotide/metabolism , Leukemia, Myeloid, Acute/genetics , Antigens, CD/immunology , Antigens, CD34/immunology , Antigens, Differentiation, Myelomonocytic/immunology , Biomarkers , Flow Cytometry , Humans , Immunophenotyping , Leukemia, Myeloid, Acute/immunology , Nucleic Acid Conformation , SELEX Aptamer Technique , Sialic Acid Binding Ig-like Lectin 3
20.
Zhongguo Shi Yan Xue Ye Xue Za Zhi ; 19(2): 332-6, 2011 Apr.
Article in Chinese | MEDLINE | ID: mdl-21518483

ABSTRACT

Constitutive activation of nuclear transcription factor-κB (NF-κB) exists in a variety of leukemia, and induction of apoptosis through blocking NF-κB activation may be an alternative strategy for leukemia treatment. The aim of this study was to investigate the inducing effect of modified adenovirus 5-based adenovirus vector (i.e. chimeric Ad5F35 Vec)-mediated expression of mutant IκBα (IκBαDN) on apoptosis of HL-60 cells. The recombinant Ad5F35-IκBαDN Vec carrying IκBαDN cDNA which deleted the first 1-70 amino acids coding sequences at 5' terminal of human IκBα was transfected into HL-60 cells. The apoptosis, NF-κB DNA binding activity, the expressions of IκBα, cIAP-2 and xIAP in HL-60 cells were detected by DNA binding assay, flow cytometry, real-time quantitative polymerase chain reaction and Western blot respectively. The results showed that apoptosis rates were 22.53 ± 2.999%, 6.08 ± 2.464% and 4.86 ± 1.366% for Ad5F35-IκBαDN Vec-infected or blank vector of Ad5F35-EGFP Vec-transfected and untransfected HL-60 cells respectively, which showed a significant difference between Ad5F35-IκBαDN Vec-transfected and untransfected cells (p < 0.001) and between Ad5F35-IκBαDN Vec-transfected and Ad5F35-EGFP Vec-transfected cells (p < 0.001, p < 0.002), while NF-κB DNA binding activity was decreased, the truncated IκBα was expressed, and IκBα mRNA expression was up-regulated, but the expression of cIAP-2 and xIAP mRNA was down-regulated after transduction for 48 hours. It is concluded that the chimeric Ad5F35 Vec can effectively mediate the expression of IκBαDN cDNA in HL-60 cells, leading to the inhibition of NF-κB DNA binding activity and inducing apoptosis of HL-60 cells.


Subject(s)
Apoptosis , I-kappa B Proteins/genetics , NF-kappa B/genetics , Adenoviridae/genetics , Genetic Vectors , HL-60 Cells , Humans , NF-KappaB Inhibitor alpha , Transfection
SELECTION OF CITATIONS
SEARCH DETAIL
...