1.Effect of Ligusticum wallichii-containing serum on expressions of Toll-like receptor 4 and myeloid differentiation factor 88 in hepatic stellate cells.
Hai-lan WANG ; Juan HE ; Wen-fu CAO ; Wen-long CHEN
China Journal of Chinese Materia Medica 2015;40(11):2191-2194
To observe the effect of Ligusticum wallichii-containing serum on the expressions of Toll-like receptor 4 and myeloid differentiation factor 88 in hepatic stellate cells. Clean-grade SD rats were randomly divided into 5 groups and orally given L. wallichii decoction, colchicine and normal saline for 7 d to prepare L. wallichii-containing serums. Except for the blank group, all of the remaining groups were stimulated with LPS 1 mg x L(-1) for 24 h. After being intervened, the L. wallichii-containing serums were cultured in 5% CO2 incubator at 37 degrees C for 24 hours. The expression of TLR4 and MyD88 were detected by RT-PCR and Western blot. After HSC was stimulated with LPS, TLR4 and MyD88 mRNA and protein expressions were significantly higher than the blank control group (P < 0.01). After being intervened with L. wallichii-containing serum, TLR4 and MyD88 mRNA and protein expressions were notably lower than the model group (P < 0.05 or P < 0.01). In conclusion, L. wallichii-containing serum could regulate the TLR4 signaling pathway and show the anti-fibrosis effect by inhibiting the expression of TLR4 and MyD88 in LPS-induced HSCs.
Animals
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Female
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Hepatic Stellate Cells
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drug effects
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metabolism
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Ligusticum
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Lipopolysaccharides
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pharmacology
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Liver Cirrhosis, Experimental
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drug therapy
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Myeloid Differentiation Factor 88
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genetics
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physiology
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Phytotherapy
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RNA, Messenger
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analysis
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Rats
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Rats, Sprague-Dawley
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Toll-Like Receptor 4
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genetics
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physiology
2.Advances in the study of gene chip technology for the investigation of the mechanisms underlying cerebral ischemia and anti-cerebral ischemia agents.
Acta Pharmaceutica Sinica 2007;42(8):803-809
With the development of molecular biology, genome science becomes an important subject currently. Characterized by high-throughput, high-integration, high-parallelism, miniaturization and automation, it is the integrated study of gene properties on a large scale. Stroke, an important cerebral vascular disease, is one of the threats to human health. The utilization of microarray study for the pathogenesis of stroke, not only reveals the essentials of the disease in the overall level of genes, but also contributes to the detection of therapeutic targets and the development of novel drugs for stroke. Referring to our own work, this discussion focuses on the progress of the mechanisms underlying experimental cerebral ischemia investigation in vivo as well as anti-cerebral ischemia agents by gene chip technology.
Animals
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Brain
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blood supply
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Brain Ischemia
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genetics
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metabolism
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Drugs, Chinese Herbal
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pharmacology
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Gene Expression Profiling
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Gene Expression Regulation
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Humans
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Hypoxia-Inducible Factor 1
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metabolism
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Interleukin-6
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metabolism
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Ischemic Preconditioning
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Neovascularization, Physiologic
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drug effects
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Neuroprotective Agents
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pharmacology
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Oligonucleotide Array Sequence Analysis
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methods
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Reperfusion Injury
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genetics
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metabolism
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Stroke
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genetics
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metabolism
3.Clinical and molecular study on Fechtner syndrome--case report and literature review.
Hai-Yan YANG ; Zhao-Yue WANG ; Yan-Hua SU ; Li-Juan CAO ; Xia BAI ; Chang-gen RUAN
Chinese Journal of Hematology 2007;28(3):160-164
OBJECTIVETo identify clinical and laboratory abnormalities and genetic defect of Fechtner syndrome in a Chinese family.
METHODSThe characteristic morphological features of platelets and leukocytes were examined on blood smears with Wright's-Giemsa staining and ultrastructure of platelet and leukocyte were investigated under electron microscope. Genomic DNA was isolated from peripheral blood of the proband and 9 members of his family. All the exons and exon-intron boundaries of the MYH9 gene were amplified by PCR followed by direct sequencing.
RESULTSPatients presented the characteristic clinical features including macrothrombocytopenia, leukocyte inclusions and/or hereditary nephritis. A heterozygous C to T mutation was found in the proband and three members of his family at nucleotide 5981 in exon 40 of MYH9 gene, resulting in a nonsense mutation which encoded truncated protein due to premature termination at the Arg 1933 codon.
CONCLUSIONIt is the first report of a Chinese family with Fechtner syndrome. The Arg (CGA) 1933--> stop (TGA) nonsense mutation in MYH9 gene is a causative genetic defect.
Adult ; Codon, Nonsense ; DNA Mutational Analysis ; Exons ; genetics ; Humans ; Inclusion Bodies ; genetics ; Male ; Molecular Motor Proteins ; genetics ; Myosin Heavy Chains ; genetics ; Nephritis, Hereditary ; genetics ; Pedigree ; Syndrome ; Thrombocytopenia ; genetics
4.Gene analysis of five inherited factor V deficiency cases.
Li-Juan CAO ; Zhao-Yue WANG ; Yan-Hua SU ; Hai-Yan YANG ; Xiao-Juan ZHAO ; Wei ZHANG ; Zi-Qiang YU ; Xia BAI ; Chang-Geng RUAN
Chinese Journal of Hematology 2008;29(3):145-148
OBJECTIVETo identify gene mutations involved in five cases of inherited factor V (FV) deficiency.
METHODSActivity of FV was determined by one-stage clotting assay using FV-deficiency plasma, and FV antigen by an ELISA assay. All the exons and exon-intron boundaries of FV gene were amplified by PCR and then DNA sequencing. Restriction enzyme analysis was used to analyze the probands, their family members and healthy volunteers.
RESULTSBoth activity and antigen of FV in the 5 patients were extremely lower compared with that of normal mixed plasma. Six mutations were identified in these 5 patients, G69969T (G2079V), C45533T (R712Ter), C46796T (R1133Ter), G45366A (C656Y), C46253T (R952C) and G16088C (D68H), the latter three were novel mutations reported for the first time and the C46253T (R952C) was the first missense mutation reported in B domain. The result of sequencing or restriction enzyme analysis showed that the three novel missense mutations were not caused by single nucleotide polymorphisms.
CONCLUSIONGene mutations in 5 type I inherited FV deficiency of patients including 2 nonsense mutations and 4 missense mutations identified which led to the instability of FV protein and the reducing of FV: Ag in the plasma.
Adolescent ; Adult ; Child ; DNA Mutational Analysis ; Exons ; genetics ; Factor V ; genetics ; metabolism ; Factor V Deficiency ; blood ; genetics ; Female ; Humans ; Male ; Mutation ; Pedigree ; Phenotype ; Young Adult
5.Expression and function of non-muscle myosin-IIA in Fechtner syndrome.
Hai-Yan YANG ; Zhao-Yue WANG ; Li-Juan CAO ; Xiao-Juan ZHAO ; Xia BAI ; Chang-Geng RUAN
Journal of Experimental Hematology 2008;16(4):871-874
The study was purposed to investigate the expression and function of non-muscle myosin heavy chain-IIA (NMMHC-IIA) in Fechtner syndrome in order to explore the pathologic changes of kindy disease and the mechanism of granulocyte inclusion body formation. NMMHC-IIA levels in granulocytes were analyzed by Western-blot, the expressions of NMMHC-IIA, IIB in HEK-293 cells were detected by RT-PCR and were analyzed by co-immunoprecipitation. The results indicated that the IIA/beta-actin ratio for Fechtner syndrome granulocytes was (0.35 +/- 0.12), and obviously decreased as compared with that of normal control (0.87 +/- 0.18) (p < 0.01). The IIA and IIB expressed higher in HEK-293 cells. The interaction of IIA and IIB was confirmed by co-immunoprecipitation in HEK-293 cells. It is concluded that dominant-negative effect of NMMHC-IIA is involved in the formation of inclusion bodies. IIA and IIB show obvious interaction, IIB partly compensates the IIA defect derived from MYH9 mutations, and may delay or prevent the development of clinically relevant abnormalities.
Blood Platelet Disorders
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genetics
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metabolism
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pathology
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Cell Line
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Granulocytes
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pathology
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Humans
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Inclusion Bodies
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pathology
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Kidney
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cytology
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embryology
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metabolism
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Mutation
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Nonmuscle Myosin Type IIA
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genetics
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metabolism
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physiology
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Nonmuscle Myosin Type IIB
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genetics
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metabolism
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physiology
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Syndrome
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Thrombocytopenia
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genetics
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metabolism
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pathology
6.Proteus syndrome with a giant hemangiomas in the spleen associated with chronic DIC--two case report and literature review.
Zhao-Yue WANG ; Yan-Hua SU ; Hai-Yan YANG ; Zi-Qiang YU ; Li-Juan CAO ; Xiao-Juan ZHAO ; Hao HU ; Sheng-Hua ZHAN ; Chang-Geng RUAN
Chinese Journal of Hematology 2007;28(3):152-155
OBJECTIVETo investigate the clinical manifestations, pathologic features and laboratory findings in two Proteus syndrome patients with giant hemangiomas in the spleen and chronic DIC.
METHODSUltrasound imaging and magnetic resonance imaging (MRI) were used for analysing the characteristics of the giant hemangiomas in the spleen. The spleen specimen was examined pathologically for the feature of the hemangioma. Homostatic tests were performed by routine laboratory methods.
RESULTSTwo Proteus syndrome patients with giant hemangiomas in the spleen causing chronic DIC (Kasabach-Merritt syndrome) were first reported. They were recovered after splenectomy.
CONCLUSIONProteus syndrome when accompanied giant hemangioma could cause chronic DIC. Significantly decreased plasma fibrinogen level in this case might be helpful for the differential diagnosis from DIC caused by other diseases.
Adolescent ; Disseminated Intravascular Coagulation ; etiology ; Female ; Hemangioma, Cavernous ; complications ; diagnostic imaging ; surgery ; Humans ; Proteus Syndrome ; complications ; Splenectomy ; Splenic Neoplasms ; complications ; diagnostic imaging ; surgery ; Ultrasonography
7.Expression of platelet membrane glycoproteins in pediatric idiopathic thrombocytopenic purpura and its clinical significance
Na WANG ; Yi-Huan CHAI ; Hai-Long HE ; Jian-Qin LI ; Lan CAO ; Li-Juan CAO
Chinese Journal of Applied Clinical Pediatrics 2013;28(3):210-213
Objective To explore the relationship between the expression of platelet membrane glycoprotein in pediatric idiopathic thrombocytopenic purpura (ITP) and its clinical significance.Methods A modified monoclonal antibody immobilization of platelet antigen (MAIPA) method was used to detect the positive expression rates of 4 platelet membrane glycoproteins (GP Ⅰ b/Ⅸ,GP Ⅰ b,GP Ⅲ a,and GP Ⅰ b) in 80 pediatric patients with ITP.The correlation was explored between the GP positive rate and the clinical efficacy in pediatric ITP.Trying to observe the correlationship between the GP positive rate of pediatric ITP in the total,the different gender,the acute and chronic and the treatment response rate in pediatric ITP respectively.Results There was a significant difference in curative rate statistically between the GP positive group and the GP negative group(x2 =8.535,P < 0.01) in 80 pediatric ITP patients,but no statistic difference in curative rate existed between the 36 female and 44 male(x2 =0.013,P >0.05).Markedly statistic difference was found in the female(x2 =4.433,P < 0.05),the same to the male (x2 =4.156,P < 0.05).Meanwhile,there was an extremely statistic difference between 67 acute and 13 chronic patients(x2 =23.513,P < 0.001).Apparently statistic difference also occurred in the acute (x2 =4.157,P < 0.05),but not in the chronic cases (x2 =0.410,P > 0.05).Conclusions The clinical response rate is significantly correlated with the GP positive rate in pediatric ITP,but not correlated with gender.The GP positive rate can reflect the disease status of pediatric ITP to a certain extent and be used as an indicator for judging the efficacy and monitor prognosis of pediatric ITP.
8.Expression of cell cycle control genes in myelodysplastic syndromes.
Jun SHI ; Zong-hong SHAO ; Hong LIU ; Hai-rong JIA ; Juan SUN ; Jie BAI ; Yan-ran CAO ; Xiu-li WANG ; Mei-feng TU
Chinese Journal of Hematology 2005;26(1):10-14
OBJECTIVETo study aberrant expression of cell cycle control genes in patients with myelodysplastic syndromes (MDS).
METHODSReverse transcription polymerase chain reaction (RT-PCR) was used to investigate the expression of cell cycle control genes (cyclin D2, cyclin D3, cyclin A1, cyclin E, CDK2, CDK4, CDK6, p21, p27, p57, Rb and E2F1) in bone marrow mononuclear cells (BMMNCs) from 29 normal control, 27 MDS and 19 de novo acute myeloid leukemia (AML).
RESULTSThe expression levels of cyclin D3 (0.65 +/- 0.17, P < 0.05) and cyclin A1 (0.48 +/- 0.04, P < 0.05) in MDS were higher than those in normal control and significantly lower than those in AML. The expression rates and levels of cyclin D2 (40.7% and 0.78 +/- 0.21) and cyclin E (51.9% and 0.52 +/- 0.10) in MDS were statistically higher than those in normal control and AML. The expression level of CDK2 in MDS (0.66 +/- 0.19, P < 0.01) was higher than that in normal control (0.42 +/- 0.04) and the expression rate of CDK6 in MDS (25.9%) higher than in normal control (3.4%, P < 0.05). There was no significant difference of the expression rates and levels of CDK4 in MDS, AML and normal control. The expression rates and levels of p21 (77.8% and 1.18 +/- 0.21) and p27 (48.1% and 1.14 +/- 0.40) in MDS were statistically higher than those in normal control and AML. The expression level of p57 in MDS (0.69 +/- 0.06) was higher than that (0.53 +/- 0.05, P < 0.01) in normal control but lower than in AML (0.96 +/- 0.16, P < 0.01). The expression rate (55.6%) and level (0.85 +/- 0.17) of Rb in MDS were significantly higher than those in normal control and AML. The expression rate (7.4%) and level (0.39 +/- 0.04) of E2F1 in MDS were comparable to those in normal control but lower than those in AML.
CONCLUSIONMDS clones have aberrant mechanism of cell cycle control: high expressions of cyclin family members, CDK2 and CDK6 may lead to high proliferation; high expression of p21 and p27 may cause the G1 phase arrest.
Adolescent ; Adult ; Cell Cycle Proteins ; genetics ; Child ; Cyclin-Dependent Kinase Inhibitor Proteins ; genetics ; Cyclin-Dependent Kinases ; genetics ; E2F1 Transcription Factor ; genetics ; Female ; Gene Expression Profiling ; Humans ; Male ; Middle Aged ; Myelodysplastic Syndromes ; genetics ; pathology ; Retinoblastoma Protein ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Young Adult
9.Quantitative and functional changes of T helper cell subsets in the bone marrow of severe aplastic anemia patients.
Guang-sheng HE ; Zong-hong SHAO ; Hong HE ; Hong LIU ; Jie BAI ; Jun SHI ; Yan-ran CAO ; Mei-feng TU ; Juan SUN ; Hai-rong JIA ; Chong-li YANG
Chinese Journal of Hematology 2004;25(10):613-616
OBJECTIVETo evaluate the quantitative and functional changes of T helper (Th) cell subsets in the bone marrow of severe aplastic anemia (SAA) patients and the relationship between these changes and the patients hematopoietic function.
METHODSBy FACS, the quantity and ratio of Th1 and Th2 cells, the percentage of CD3(+)CD8(+) cells in the bone marrow were detected in 24 patients with SAA at active phase, 15 patients with SAA at recovery phase, and 16 normal controls. By radioimmunoassay, the serum levels of TNF-alpha, or IL-4 in 20 SAA patients at active phase, 12 at recovery phase and 16 normal controls were measured. The relationships between CD3(+)CD8(+) cells, TNF-alpha and Ret, ANC; and between Th1 cells and CD3(+)CD8(+) cells, TNF-alpha or Ret, ANC; between IL-4, balance of Th1/Th2 and Ret, ANC were evaluated.
RESULTSThe percentages of Th1 and Th2 cells, and ratio of Th1/Th2 in bone marrow of SAA patients at active phase were (4.87 +/- 2.64)%, (0.41 +/- 0.26)% and 21.22 +/- 5.07, respectively, being higher than those of normal controls [(0.42 +/- 0.30)% (P < 0.01), (0.24 +/- 0.17)% (P < 0.05) and (1.57 +/- 0.93) (P < 0.01), respectively] and all of them reduced to normal levels of SAA at recovery phase (P > 0.05). The percentage of CD3(+)CD8(+) cells significantly decreased from (32.32 +/- 8.69)% at active phase to (13.76 +/- 2.96)% at recovery phase (P < 0.01). The serum levels of TNF-alpha and IL-4 at active phase was (4.29 +/- 3.15) microg/L and (1.24 +/- 0.73) microg/L, respectively, being higher than those of normal controls (1.21 +/- 1.16) microg/L, (1.18 +/- 0.97) microg/L, but only the difference of TNF-alpha was statistically significant (P < 0.01). In recovery SAA patients, the serum levels of TNF-alpha significantly decreased to (1.46 +/- 1.41) microg/L (P < 0.01), and the levels of IL-4 increased markedly to (3.05 +/- 1.94) microg/L. The CD3(+)CD8(+) cells and TNF-alpha of patients negatively correlated with Ret (P < 0.05; P < 0.05) and ANC (P < 0.05; P < 0.05), Th1 cells correlated with CD3(+)CD8(+) cells and TNF-alpha positively (P < 0.01; P < 0.05), the Ret and ANC negatively (P < 0.01; P < 0.01), IL-4 and the balance of Th1/Th2 positively correlated with Ret and ANC (P < 0.05, P < 0.01; P < 0.01, P < 0.01).
CONCLUSIONThe bone marrow failure in SAA might be caused not only by the increase of Th1 cells, Th1 type effector cells and cytokines, but also by insufficient compensation of Th2 cells and Th2 type cytokines, which shifted the balance of Th1/Th2 favorable to Th1.
Adolescent ; Adult ; Anemia, Aplastic ; blood ; pathology ; physiopathology ; Bone Marrow ; metabolism ; pathology ; CD3 Complex ; blood ; CD8 Antigens ; blood ; Child ; Female ; Hematopoietic System ; metabolism ; pathology ; physiopathology ; Humans ; Interleukin-4 ; blood ; Male ; Middle Aged ; Radioimmunoassay ; T-Lymphocytes, Helper-Inducer ; metabolism ; pathology ; Th1 Cells ; metabolism ; pathology ; Th2 Cells ; metabolism ; pathology ; Tumor Necrosis Factor-alpha ; blood ; Young Adult
10.Changes of subsets of DC1 in the bone marrow of severe aplastic anemia patients.
Guang-sheng HE ; Zong-hong SHAO ; Hong HE ; Hong LIU ; Rong FU ; Jie BAI ; Jun SHI ; Yan-ran CAO ; Mei-feng TU ; Juan SUN ; Hai-rong JIA
Chinese Journal of Hematology 2004;25(11):649-652
OBJECTIVETo measure the subsets of dendritic cells 1 (DC1) in the bone marrow of severe aplastic anemia (SAA) patients and evaluate the relationships between the CD11c+CD83+ cells and Th1 cells, CD3+CD8+ cells or hematopoietic function and explore the role of DC1 in the pathogenesis of SAA.
METHODSBy FACS, the quantities and ratios of CD11c+CD1a+ cells, CD11c+CD83+ cells, Th1 cells, and CD3+CD8+ cells in the bone marrow of SAA patients and normal controls were detected respectively. The relationships between CD3+CD8+ cells and reticulocyte absolute value (Ret) or neutrophil absolute value (ANC), between Th1 cells and CD3+CD8+ cells, Ret or ANC, between CD11c+CD83+ cells, and Th1 cells, CD3+CD8+ cells, Ret or ANC were evaluated.
RESULTSIn normal controls' bone marrow, the percentages of Th1 cells, CD11c+CD1a+ cells, CD11c+CD83+ cells and the ratio of CD11c+CD83+/CD11c+CD1a+ were (0.42 +/- 0.30)%, (0.38 +/- 0.29)%, (0.37 +/- 0.32)% and 1.07 +/- 0.10, respectively. In untreated SAA patients, they were (4.87 +/- 0.54)%, (1.73 +/- 0.24)%, (3.38 +/- 0.56)% and 2.21 +/- 0.32 respectively, which were higher than that in normal controls (P < 0.01). In recovering SAA patients, the percentages of Th1 cells, CD11c+CD1a+ cells and CD11c+CD83+ cells decreased significantly to (0.53 +/- 0.22)%, (0.61 +/- 0.23)%, (0.65 +/- 0.22)%, respectively (P < 0.01). The ratio of CD11c+CD83+/CD11c+ CD1a+ in recovering SAA patients decreased to 1.37 +/- 0.25, which was similar to that in normal controls (P > 0.05). The percentage of CD3+CD8+ cells in untreated SAA patients was (32.32 +/- 10.22)%, and in recovering SAA patients decreased to (13.67 +/- 5.24)% (P < 0.01). The percentage of CD3+CD8+ cells in SAA patients was negatively correlated with their Ret and ANC (P < 0.05), while their Th1 cell percentages were positively correlated with their CD3+CD8+ cells (P < 0.01), and negatively correlated with their Ret and ANC (P < 0.01). SAA patient's CD11c+CD83+ cell percentages were positively correlated with their Th1 cell and CD3+CD8 cells (P < 0.01, P < 0.05), but negatively with their Ret and ANC (P < 0.01).
CONCLUSIONBoth immature DC1 and activated DC1 increased in the bone marrow of SAA patients, and the balance of DC1 subsets shifted from stable form to active one, which might promote Th0 cells to polarize to Th1 cells, and cause the over-function of T lymphocytes and hematopoiesis failure in SAA.
Adolescent ; Adult ; Anemia, Aplastic ; immunology ; Antigens, CD ; immunology ; Antigens, CD1 ; immunology ; Bone Marrow ; immunology ; CD11c Antigen ; immunology ; CD8-Positive T-Lymphocytes ; immunology ; Child ; Dendritic Cells ; immunology ; Female ; Humans ; Immunoglobulins ; immunology ; Male ; Membrane Glycoproteins ; immunology ; Th1 Cells ; immunology ; Young Adult