1.Effect of soy isoflavone on incretion of female rats
Xingming ZHONG ; Yiping ZHONG ; Quan QI ; Zhulin MIAO ; Rong CUI ; Xiaolan WANG ; Miqiong HUANG
Chinese Journal of Primary Medicine and Pharmacy 2009;16(1):75-77
Objective To observe the effect of soy isoflavones on incretion of female rats. Methods 42 fe-male rots of 12 weeks old were divided into 3 groups at random. Low dose group were perfused with 40mg · kg-1·d-1 into stomach per day;high dose group were perfused with 80mg·kg-1·d-1 into stomach per day;control group were perfused with physiological saline into stomach. After 14 days,collected blood via jugular vein and tested 4 inde-xes-FSH, LH,E2 and P with chemoluminescence method. Results In 3 groups of soy isoflavones low doee group,soy isoflavones high dose group and control group,FSH were (0.13±0.021) mIu/ml, (0.12±0.018) mIu/ml, (0.15 ±0.024) mIu/ml respectively; LH were (0.17±0.032) mIu/ml, (0.15±0.043) mIu/ml, (0.18±0.047) mIu/ml respectively, which has no obvious differences (P > 0.05) ; E2 were (0.09±0.03) nmol/L, (0.03±0.03) nmol/L, (0.12±0.04) nmol/L respectively; P were (1.43±0.27) ng/ml, (2.82±0.37) ng/ml, (0.67±0.56) ng/ml re-spectivdy. Compare those 3 groups, E2 activeness of soy isoflavones group decreased obviously; but P activeness of soy isoflavones group increased obviously. Conclusion Soy isoflavones has no obvious effect on hypophysis hormone of rat, but the soy isoflavones of different dosages may measure the secretion of female rat ovary hormones by estrogen ac-tivity and antiestrogen activity.
2.Effects of storage time on quality of Desmodium styracifolium seeds.
Quan YANG ; Xiao-min TANG ; Hai-yun PAN ; Ling-feng MEI ; Chun-rong ZHANG ; Xuan-xuan CHENG ; Lu-qi HUANG
China Journal of Chinese Materia Medica 2015;40(20):3953-3957
The dynamic changes of germination percentage, germination potential, thousand-seed weight, antioxidase activity in Desmodium styracifolium seeds with different storage time were tested, and electrical conductivity, contents of soluble sugar, soluble protein, starch in seed leach liquor were also determined in order to reveal the mechanism of seed deterioration. The results as the following. (1) The germination percentage, germination potential and thousand-seed weight of D. styracifolium seeds declined, while the seed coat color darkened with the extension of storage time. (2) The activities of superoxide dismutase (SOD) and peroxidase (POD) decreased with the prolongation of storage period. The SOD activity declined fastest in 1,095-1,185 d of storage, while the POD activity declined significantly in 365-395 d of storage. (3) The electrical conductivity and the contents of soluble sugar, starch in seed leach liquor increased, while the content of soluble protein declined with the extension of storage time. (4) Correlation analysis indicated that the germination percentage, germination potential and thousand-seed weight of D. styracifolium seeds have a significantly positive correlation with SOD and POD activity, while have a significantly negative correlation with the electrical conductivity, contents of soluble sugar and starch. It can be concluded that during the storage of D. styracifolium seeds, physiological and biochemical changes including decrease in antioxidase activity, rise in electrical conductivity, degradation effluent of soluble sugar and starch, degradation of soluble protein were the main factors leading to the seed deterioration.
Color
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Fabaceae
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chemistry
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enzymology
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growth & development
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metabolism
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Germination
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Peroxidases
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metabolism
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Plant Proteins
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metabolism
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Seeds
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chemistry
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enzymology
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growth & development
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metabolism
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Starch
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metabolism
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Superoxide Dismutase
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metabolism
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Time Factors
3.Role of matrix metalloproteinase (MMP)-2 and -9 and vascular endothelial growth factor C in lymph node metastasis of breast cancer.
Shao-qiu ZHENG ; Rong-quan HUANG ; Ya-jie ZHANG
Chinese Journal of Pathology 2010;39(4):240-244
OBJECTIVETo study the expression of matrix metalloproteinase (MMP)-2, MMP-9 and vascular endothelial growth factor (VEGF)-C in breast cancer and their role in lymph node metastasis.
METHODSImmunohistochemical staining was used to detect the expression of VEGF-C, MMP-2, MMP-9 and LYVE-1 in 84 cases of breast cancer, including 52 cases with and 32 cases without lymph node metastases. The recombinant vector (pSIREN-VEGF-C) was transfected into human breast cancer cell MCF-7 by liposome, and the RNA expression of MMP-2, MMP-9 and VEGF-C in MCF-7 cells after transfection was detected by PCR.
RESULTSThe expressions of MMP-2, MMP-9 and VEGF-C were 98.1% (51/52), 88.5% (46/52), and 94.2% (49/52) respectively for the metastatic group, and 75.0% (24/32), 53.1% (17/32), and 65.6% (21/32) respectively for the non metastatic group, and there was significant difference between these groups (P < 0.05). The lymphatic vessel density between these two groups was also significantly different (P < 0.05). Increased expression of MMP-2, MMP-9 and VEGF-C was also associated with increased number of lymphatic vessels had also increased (P < 0.05). The expression of MMP-2, MMP-9 and VEGF-C in MCF-7 cells after gene transfection decreased significantly (P < 0.05).
CONCLUSIONMMP-2 and MMP-9 in conjunction with VEGF-C, promote lymphangiogenesis and lymph node metastasis of breast cancer.
Adult ; Aged ; Breast Neoplasms ; genetics ; metabolism ; pathology ; surgery ; Carcinoma, Ductal, Breast ; genetics ; metabolism ; pathology ; surgery ; Carcinoma, Medullary ; genetics ; metabolism ; pathology ; surgery ; Cell Line, Tumor ; Female ; Humans ; Lymph Nodes ; metabolism ; pathology ; Lymphangiogenesis ; Lymphatic Metastasis ; Lymphatic Vessels ; metabolism ; pathology ; Matrix Metalloproteinase 2 ; genetics ; metabolism ; Matrix Metalloproteinase 9 ; genetics ; metabolism ; Middle Aged ; RNA Interference ; RNA, Messenger ; metabolism ; RNA, Small Interfering ; genetics ; Recombinant Proteins ; genetics ; metabolism ; Transfection ; Vascular Endothelial Growth Factor C ; genetics ; metabolism ; Vesicular Transport Proteins ; metabolism
4.Experimental study of the inhibitory effects of biotic royal jelly on the growth of ascitic hepatoma cell H22 in mice
Yong-Zi WANG ; Chang-Quan LING ; Xue-Qiang HUANG ; Pin-Rong PAN
Academic Journal of Second Military Medical University 2001;22(4):357-359
Objective: To observe the inhibitory effects of biotic royal jelly on the ascitic hepatoma cell H22. Methods: Mice bearing H22 tumor were fed on different types of royal jelly: No.1, 2 and 3. Their anti-tumor effects were observed in vivo. The general royal jelly and normal saline were observed as control. Results: Among these biotic royal jelly, the biotic royal jelly No.1 showed obvious tumor-inhibiting and survival-prolonging effects. In addition, it increased the number of WBC and augmented the amount of IL-2 and IFN-γ; the pathological study also indicated the denaturing and necrosis of most tumor cells with nuclei constraining and cell membrane rupturing, and large amount of lymphocytes and plasmacytes infiltrating around the mass. Conclusion: The No.1 biotic royal jelly has obvious anti-tumor effect, and it may take effects by inhibiting or killing tumor cells and improving the immunity of the host.
5.The Analysis of an Inquiry in Grouping Teaching among Medical Students——the Research of Grouping Teaching Model,Students Inquiry
Li-Ping CHEN ; Zhi-Quan BAI ; Jiu-Xiang LI ; Yu-Zhuang HUANG ; Jing-Rong ZENG ; Jing-Bo JIANG ;
Chinese Journal of Medical Education Research 2006;0(10):-
By inquiring the medical students under the background of grouping teaching between the mainland students and the oversea Chinese students,we have got something about their attitude toward the credit system.The result will help us to improve the teaching renovation in medical education.The questionnaire including implementing of credit system,standard credit system, grouping teaching,curriculum,tutor system of the undergraduates,the administration of education,and so on.Then we analyze and get the result.
6.Gfi-1 expression in leukemia patients and inhibitory effects of lentiviral vector mediated silence of Gfi-1 gene on proliferation in K562 cells.
Rong ZHAN ; Shun-Quan WU ; Hao-Bo HUANG ; Shen-Lin HUANG ; Jun LIN
Journal of Experimental Hematology 2010;18(4):849-854
This study was aimed to quantitatively detect the expression levels of gfi-1 gene in leukemia patients, and to investigate the effect of gfi-1 gene silenced by short hairpin RNA (shRNA) on proliferation of leukemia cell line K562. Quantitative real-time PCR and Western blot were used to detect the mRNA and protein expression levels of GFI-1 in newly diagnosed patients with leukemia. One pair of oligonucleotide sequences targeted at human gfi-1 mRNA were designed and synthesized. The annealed oligonucleotide fragments were subcloned into pLVTHM vector. Virus particles were collected when the control and shRNA vectors had been co-transfected with the psPAX2 packaging plasmid and the envelope plasmid pMD2 G into HEK-293T cells using Lipofectamine 2000. The K562 cells were transfused with 1 x 10⁶ recombinant lentivirus-transfusing units plus 6 microg/ml of polybrene. Rea-time PCR and Western blot were used to detect the expressions of gfi-1 and bax mRNA after lentivirus transfusion. CCK-8 assays was used to evaluate the proliferation potential of cells. The results showed that the gfi-1 expression level in all leukemia patients was significantly higher than that in normal group (p < 0.05); the gfi-1 mRNA expression in chronic myeloid leukemia (CML) and acute myeloid leukemia (AML) or acute lymphoid leukemia (ALL) patients was significantly higher than that in normal group (p < 0.05); but the difference of gfi-1 mRNA expression between AL and CML or ALL and AML was not significant. Notably, the gfi-1 mRNA expression level had a positive correlation with high white blood cell count of > 20.0 x 10⁹/L (p < 0.05). As was expected, the mRNA and protein level of gfi-1 was reduced significantly in K562 cells after lentivirus transfusion, whereas the mRNA and protein level of bax was upregulated. And CCK-8 assay showed that gfi-1 gene silencing can inhibit K562 proliferation. It is concluded that gfi-1 expression is upregulated in leukemia patients and may contribute to leukemogenesis. The gfi-1 specific shRNA mediated by lentivirus can effectively down-regulate the expression of gfi-1 and inhibit the proliferation of K562 cells, which lay a basis for further research on gene therapy in leukemia.
Adolescent
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Adult
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Aged
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Case-Control Studies
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Cell Proliferation
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DNA-Binding Proteins
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genetics
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Female
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Gene Silencing
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Genetic Vectors
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Humans
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K562 Cells
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Lentivirus
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genetics
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Leukemia
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genetics
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pathology
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Male
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Middle Aged
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Plasmids
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RNA, Small Interfering
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genetics
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Transcription Factors
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genetics
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Young Adult
7.Identification of herbal tea ingredient Plumeria rubra and its adulterants using DNA barcoding.
Yu-Hua SHI ; Wei SUN ; Guang-Hong FANG ; Rong-Bo ZHENG ; Wen-Liu XU ; Xiao-Dan HUANG ; Shao-Quan WENG ; Chu-Yuan LI ; Shi-Lin CHEN
China Journal of Chinese Materia Medica 2014;39(12):2199-2203
ITS2 sequence was used as a barcode to identify herbal tea ingredient Plumeria rubra and its adulterants. Genomic DNAs from forty eight samples were extracted, the ITS2 sequences were amplified and sequenced bi-direstionlly, and then assembled and obtained using CodonCode Aligner. The sequences were aligned using ClustalW, the genetic distances were computed by kimura 2-parameter (K2P) model and the Neighbor-joining (NJ) phylogenetic trees were constructed using MEGA5.0. Results showed that the length of ITS2 sequence of P. rubra were 244 bp. The intra-specific genetic distances (0-0. 016 6) were much smaller than inter-specific ones between P. rubra and its adulterants(0.320 8-0.650 4). The NJ tree indicated that P. rubra and its adulterants could be distinguished clearly. Therefore, Using ITS2 barcode can accurately andeffectively distinguish herbal tea ingredient P. rubra from its adulterants, which providesa new molecular method to identify P. rubra and ensure its safety in use.
Apocynaceae
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classification
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genetics
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DNA Barcoding, Taxonomic
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methods
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DNA, Plant
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genetics
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DNA, Ribosomal Spacer
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genetics
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Drug Contamination
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prevention & control
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Drugs, Chinese Herbal
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chemistry
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classification
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Flowers
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chemistry
;
classification
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Molecular Sequence Data
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Phylogeny
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Quality Control
8.Establishment and application of lentivirus luciferase reporter-mediated miRNA target gene screening system.
Shun-Quan WU ; Jun LIN ; Sheng-Lin HUANG ; Rong ZHAN
Journal of Experimental Hematology 2012;20(1):159-163
This study was aimed to establish a high-throughput luciferase reporter system, through which to screen and identify miRNAs directly targeting p21, and to explore the biological function and significance of these miRNAs. Molecular cloning technique was used to construct and identify two lentivirus-expressing vectors-pWPXL-Luc and pWPXL-Luc-P21-3'UTR, virus particles were collected after the pWPXL-Luc or pWPXL-Luc-P21-3'UTR vectors were co-transfected with the psPAX2 packaging plasmid and the envelope plasmid pDM2G into HEK-293T cells. Furthermore, two stable cell lines expressing luciferase singly or co-expressed luciferase and P21-3'UTR were established by transducing HEK-293 cells with recombinant lentivirus; the former was used as control in the following experiments. Finally luciferase activity of the latter stable cells was measured by using the luciferase reporter assay system. The results showed that high-titre recombinant lentivirus was produced and two stable cell lines were constructed, also to some certain, the luciferase activity was in direct proportion to the number of cells. In conclusion, the high-throughput luciferase reporter system is established successfully; using this system, the 28 miRNA that directly target P21 Cip1/Waf1 are screened experimentally.
Genes, Reporter
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Genetic Vectors
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HEK293 Cells
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Humans
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Lentivirus
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genetics
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Luciferases
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genetics
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MicroRNAs
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genetics
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Plasmids
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Transduction, Genetic
9.Lysophosphatidic acid acyltransferase β gene expression in newly diagnosed leukemia patients.
Rong ZHAN ; Hao-Bo HUANG ; Shun-Quan WU ; Jun LIN
Journal of Experimental Hematology 2010;18(6):1422-1426
This study was aimed to quantitatively detect the expression level of lysophosphatide acid acyltransferase β (lpaat β) mRNA in leukemia patients so as to provide theoretical basis for the target therapy of lpaat β in leukemia. Real-time fluorescently quantitative PCR was used to detect the relative expression level of lpaat β mRNA to analyze its expression change in various type of leukemia. The results showed that the lpaat β mRNA expression level in acute leukemia (AL) patients was significantly higher than that in normal controls (p < 0.05); lpaat β mRNA expression level in acute myeloid leukemia (AML) patients was significantly higher than that in normal controls (p < 0.05) and was positively correlated with white blood cell count (≥ 20.0 × 10(9)/L) (p < 0.05) and CD34 expression level of leukemia, but was not related with extramedullary infiltration. Except for acute promyelocytic leukemia (APL), the lpaat β mRNA expression level was negatively correlated with chemotherapy sensitivity in chronic myeloid leukemia (AML) patients. lpaat β mRNA expression level in chronic myeloid leukemia (CML) patients was significantly higher than that in normal controls (p < 0.05). lpaat β mRNA expression level in acute lymphoid leukemia (ALL) patients was not higher than that in normal controls (p > 0.05). It is concluded that the lpaat β gene overexpression exists in both AML and CML patients. lpaat β produced by AML cells probably plays an important role in abnormal proliferation and drug-resistance of AML cells.
Acyltransferases
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genetics
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Adolescent
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Adult
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Aged
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Aged, 80 and over
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Case-Control Studies
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Child
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Female
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Gene Expression
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Humans
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Leukemia
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genetics
;
pathology
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Male
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Middle Aged
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RNA, Messenger
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genetics
;
Young Adult
10.april mRNA expression in newly diagnosed leukemia patients.
Hao-Bo HUANG ; Shun-Quan WU ; Rong ZHAN ; Li-Ping FAN
Journal of Experimental Hematology 2010;18(6):1414-1417
This study was aimed to quantitatively detect the levels of april mRNA expression in leukemia patients so as to provide theoretical basis for the target therapy directing at april in leukemia. Real time fluorescent quantitative PCR was used to detect the relative expression level of april mRNA in newly diagnosed leukemia patients and to analyze the changes of its expression level in various type of leukemia. The results showed that the april mRNA expression level in acute leukemia (AL) patients was significantly higher than that in normal controls, there was statistical difference between them (p < 0.05); april mRNA expression level in acute myeloid leukemia (AML) patients was significantly higher than that in normal controls (p < 0.05) and positively correlated with white blood cell count ≥ 20.0 × 10(9)/L (p < 0.05), but not related with extramedullary infiltration and the expression of CD34. Except for acute promyelocytic leukemia (APL), april mRNA expression level was negatively correlated with sensitivity of patients to chemotherapy. april mRNA expression levels in acute lymphoid leukemia (ALL) and chronic myeloid leukemia (CML) patients were not higher than that in normal controls, there was no statistical difference between them (p > 0.05). It is concluded that april gene overexpression exits in AML patients. APRIL protein produced by AML cells probably plays an important role in abnormal proliferation and drug-resistance of AML cells.
Adolescent
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Adult
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Aged
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Aged, 80 and over
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Case-Control Studies
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Female
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Humans
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Leukemia
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genetics
;
therapy
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Male
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Middle Aged
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RNA, Messenger
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genetics
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Tumor Necrosis Factor Ligand Superfamily Member 13
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genetics
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Young Adult