1.Effects of PRMT5 expression on cell proliferation of ovarian cancer cell HO8910
Ye WEI ; Ran-Ran WANG ; Tian-Tian HAN ; Kang-Rong ZHAO ; Qiong LIN ; Gen-Bao SHAO
Journal of Medical Postgraduates 2018;31(6):565-572
Objective Protein arginine methyltransferase 5 (PRMT5),a member of the histone arginine methylation transferase family,is involved in a wide range of biological regulation through ei-ther epigenetic or posttranslational methylation modifications. The pur-pose of the present study was to investigate the effects of PRMT5 on cell proliferation of ovarian cancer cell HO8910. Methods Cell lines HO8910 with PRMT5 overexpression were obtained by transi-ent transfection,which were divided into three groups in the experiment: blank control group (wild-type cell line HO8910),negative control group (HO8910 cells were transfected with pCMV-myc plasmid),and experimental group (HO8910 cells were transfected with pCMV-myc-PRMT5 plasmid). Western blot was used to detect the expression of myc protein,and qRT-PCR was used to detect the ex-pression of PRMT5 mRNA. Cell lines HO8910 with inducible stable knockdown of PRMT5 were established by shRNA interference method,which were divided into four groups: pLKO control group (infected by empty vector lentivirus),pLKO+Dox (100ng/mL) group,shPRMT5 group (infected by PRMT5shRNA lentivirus) and shPRMT5+Dox (100 ng/mL) group. Western blot and qRT-PCR were used to detect the expression of PRMT5 protein and mRNA levels. Dox-induced PRMT5 knockdown was detected by increasing Dox concentration,which includes four groups,Dox 0ng/mL group,Dox 1ng/mL group,Dox 10ng/mL group,Dox 100ng/mL group,and each group was treated for 48 hours. Western blot and qRT-PCR were used to detect the PRMT5 protein and mRNA expression. Colony formation assay,EdU assay,and CCK-8 assay were used to test cells proliferation. The experiment was conducted in two large groups each with two subgroups: PRMT5 knockdown group (Dox-group,Dox+ group),PRMT5 overexpression group (pCMV-myc group,pCMV-myc-PRMT5 group). Western blot was used to detect the effects of PRMT5 expression on proliferation-related proteins. The experiment was conducted in two large groups,PRMT5 knockdown group with four subgroups : Dox 0ng/mL group,Dox 1ng/mL group,Dox 10ng/mL group and Dox 100ng/mL group,and PRMT5 overexpression group with two subgroups (pCMV-myc group and pCMV-myc-PRMT5 group). Results Western blot results showed that the expression of myc was detected in the experimental group in which HO8910 cells were transfected with pCMV-myc-PRMT5,and the expression of PRMT5 mRNA was significantly higher in the experimental group than those in the blank control group and the negative control group (P<0.001) . Western blot and qRT-PCR showed that PRMT5 protein (0.32±0.25) and mRNA expression levels in shPRMT5+Dox group were significantly lower than those of shPRMT5 group (0.89±0.18) (P<0.05). Western blot and qRT-PCR confirmed that PRMT5 protein (0.21±0.24) and mRNA expres-sion in Dox 10ng/mL group and Dox 100ng/mL group (0.08±0.15) were significantly downregulated compared to Dox 0ng/mL group (1.11±0.15) (P<0.05). Colony formation experiments,EdU experiments,and CCK-8 experiments confirmed that the proliferative ca-pacity of cells in Dox+group was lower than that of Dox-group in PRMT5 knockdown group(P<0.05); while in PRMT5 overexpression group,the proliferative capacity of pCMV-myc-PRMT5 group was significantly higher than that of the pCMV-myc group (P<0.05). Western blot results showed that the protein expression of Cyclin D1 was significantly lower in Dox 100 ng/mL group (0.17±0.06) than that in Dox 0 ng/mL group (1.18±0.18) (P<0.05) and the expression of P21 was significantly increased in PRMT5 knockdown group (P<0.05). In the PRMT5 overexpression group,the protein expression of Cyclin D1 in pCMV-myc-PRMT5 group (3.48± 0.22) was higher than that in pCMV-myc group (0.88±0.15) (P<0.05),while the protein expression of P21 (0.08±0.17) were significantly lower than that of pCMV-myc group (4.12±0.10) (P<0.05). Conclusion PRMT5 plays an important role in the proliferation of ovarian cancer cells. Down-regulation of PRMT5 can inhibit cell proliferation and up-regulation of PRMT5 can pro-mote cell proliferation.
2.Study Progress of Cu,Zn Superoxide Dismutas——From Gene to Function
Chang-Lu WANG ; Jun-Wu CAO ; Yu-Rong WANG ; Mian-Hua CHEN ; Zhi-Qiang CHEN ; Shao-Ran TIAN
Progress in Modern Biomedicine 2008;8(5):940-943
Superoxide Dismutase (SOD)(EC 1.15.1.1)is a metalloenzyme that is found in almost all organisms and catalyzes the dismutation of superoxide anion radical to hydrogen peroxide and molecular oxygen. Three unique and highly compartmentalized mammalian SOD have been biochemically and molecularly characterized to date: Cu, Zn superoxide dismutase (CuZnSOD, SOD1), MnSOD (Manganese Superoxide Dismutase, SOD2)and EC-SOD (Extracellular Superoxide Dismutase, SOD3). Cu, Zn superoxide dismutase (CuZnSOD, SOD1)is a copper and zinc-containing homodimer that is found almost exclusively in intracellular cytoplasmic spaces. CuZnSOD is widely distributed and comprises about 90% of the total SOD. Cytoplasmic and periplasmic SOD exists as dimers,whereas chloroplastic and extracellular enzymes exist as tetramers. Structure supports independent functional evolution in prokaryotes and eukaryotes. CuZnSOD are thought to protect the brain, lungs, and other tissues from oxidative stress. This paper reviewed the gene, molecular and chemical structure and biological function of CuZnSOD.
3. Clinical features of dyslipidemia in patients with primary biliary cholangitis
Tihong SHAO ; Ran TIAN ; Jinlei SUN ; Shuo ZHANG ; Yihan CAO ; Zhilei CHEN ; Li WANG ; Fengchun ZHANG
Chinese Journal of General Practitioners 2018;17(8):617-620
Objective:
To analyze the clinical features of dyslipidemia in patients with primary biliary cholangitis (PBC).
Methods:
The clinical and laboratory data of 136 PBC patients in Peking Union Medical College Hospital from 2010 to 2016 were retrospectively analyzed.The liver function was compared between patients with normal and abnormal blood lipids.
Results:
Among 136 PBC patients, 100(74%)had abnormal serum lipids. The incidence of increased cholesterol, low-density lipoprotein and triglyceride was 61%(59/96), 58%(48/83) and 47%(46/97), respectively; while that of reduced HDL-C was 26%(21/82). The incidences of pruritus [26%(26/100)
4.Detection for endogenous erythroid colony in the patients with polycythemia vera and its clinical significance.
Jie BAI ; Zong-hong SHAO ; Hong LIU ; Jun SHI ; Guang-sheng HE ; Yan-ran CAO ; Zhen-zhu CUI ; Juan SUN ; Zheng TIAN ; Hai-rong JIA ; Lin-sheng QIAN ; Tian-ying YANG ; Chong-li YANG
Chinese Journal of Hematology 2003;24(11):561-564
OBJECTIVETo investigate the growth of endogenous erythroid colony (EEC) in polycythemia vera (PV) patients and its clinical significance.
METHODSBone marrow mononuclear cells of 26 PV patients, 2 secondary erythrocytosis (SE) and 19 normal controls were cultured by Marsh's method for EEC.
RESULTS1. EEC was present in 25/26 (96.2%) PV patients and was not found in 2 SE patients and 19 normal controls. 2. The number of EEC and the ratio of EEC/Epo-dependent CFU-E (EEC ratio) were positively correlated with the hemoglobin (Hb) levels (r = 0.608, P = 0.01) in PV patients, but did not correlate with white blood cell (WBC) counts, platelet counts and neutrophil alkaline phosphatase scores. 3. EEC did not correlate with PV patients' serum Epo levels (r = 0.518, P = 0.125). 4. Fifteen PV patients were treated with hydroxyurea and/or interferon-alpha. Their EEC ratio before treatment was correlated positively with the time required for complete remission (CR) (r = 0.651, P = 0.009) and negatively with the time before relapsing (r = -0.529, P < 0.02). 5. EECs of 7 PV patients treated with HU/IFN were decreased after their blood cell counts normalization. 6. There was a positive correlation between the EEC ratio and the attacks of vascular thrombosis (r = 0.524, P = 0.01). (7) The apoptosis of bone marrow mononuclear cells of PV patients was less than that of normal controls. PV patients' EEC was negatively correlated with the apoptosis of their bone marrow mononuclear cells (r = -0.192, P < 0.045).
CONCLUSIONEEC is peculiarly present in PV patients, and is a sensitive parameter in reflecting the abnormal hematopoietic clone burden and in diagnosing and monitoring the disease.
Adult ; Aged ; Apoptosis ; Bone Marrow Cells ; physiology ; Erythroid Precursor Cells ; physiology ; Erythropoietin ; blood ; Female ; Humans ; Male ; Middle Aged ; Polycythemia Vera ; blood ; therapy
5.Endogenous erythroid colony assay in patients with polycythemia vera and its clinical significance.
Jie BAI ; Zong-hong SHAO ; Hong LIU ; Jun SHI ; Guang-sheng HE ; Yan-ran CAO ; Zhen-zhu CUI ; Yu-hong WU ; Juan SUN ; Zheng TIAN ; Hai-rong JIA ; Lin-sheng QIAN ; Tian-ying YANG ; Chong-li YANG
Chinese Medical Journal 2004;117(5):668-672
BACKGROUNDPolycythemia vera (PV) is a malignant disorder of hemaopoietic stem cells which is characterized by clonal hyperproliferation and a low rate of apoptosis. This study was to assess endogenous erythroid colony (EEC) formation in the bone marrow of PV patients and determine its clinical significance.
METHODSThe bone marrow mononuclear cells of 26 patients with PV, 2 patients with secondary erythrocytosis (SE), and 19 normal controls were cultured by Marsh's method for EEC evaluation, and the clinical significance was evaluated.
RESULTSEECs appeared in 25 patients with PV but not in 2 patients with SE and 19 normal controls. The number of EECs and the EEC ratio [EEC/erythropoietin (EPO)-dependent colony forming unit-erythroid (CFU-E)] in PV patients positively correlated with hemoglobin (Hb) levels. Their EEC number did not correlate with white blood cell (WBC) counts, platelet (PLT) counts, or leukocyte alkaline phosphatase (LAP) scores. Their EEC did not correlate with serum EPO levels. Fifteen patients with PV were treated with hydroxyurea (Hu) and/or interferon-alpha (IFN-alpha). Their EEC ratio before treatment positively correlated with the treatment time required for complete remission (CR) and negatively correlated with the time before relapse. The EEC numbers of 7 PV patients treated with Hu/IFN-alpha decreased after the blood cell counts dropped to normal levels. There was a positive correlation between the EEC ratio and the incidence of attacks of vascular thrombosis in PV patients. The numbers of apoptosised bone marrow mononuclear cells in PV patients were lower than those in normal controls. The EEC numbers of PV patients negatively correlated with the rate of apoptosis of bone marrow mononuclear cells.
CONCLUSIONSEEC formation is characteristic in PV patients. EEC number in PV patients positively correlates with Hb levels, the time required for CR, and the incidence of attacks of vascular thrombosis. EEC number negatively correlates with the time before relapse. Bone marrow suppressive treatment might decrease EEC number. Thus, EEC number is a sensitive and specific parameter reflecting the abnormal hematopoietic clone burden induced by polycythemia vera. EEC number is an important diagnostic parameter for PV patients.
Adult ; Aged ; Apoptosis ; Colony-Forming Units Assay ; Erythroid Precursor Cells ; physiology ; Erythropoiesis ; Erythropoietin ; blood ; Female ; Humans ; Male ; Middle Aged ; Polycythemia Vera ; blood ; therapy ; Thrombosis ; epidemiology
6.Study on the burden of abnormal hematopoietic clone of the patients with myelodysplastic syndromes and its implications.
Hua-Quan WANG ; Zong-Hong SHAO ; Jun SHI ; Yan-Ran CAO ; Hong LIU ; Jie BAI ; Mei-Feng TU ; Li-Min XING ; Zhen-Zhu CUI ; Shi-He LIU ; Juan SUN ; Hai-Rong JIA ; Tian-Ying YANG
Chinese Journal of Hematology 2005;26(8):473-476
OBJECTIVETo investigate the abnormal hematopoietic clone burden of the patients with myelodysplastic syndromes (MDS) and its clinical implication.
METHODSThe ratio of the metaphase with abnormal karyotypes to the total was regarded as the index of MDS clonal burden. Thirteen parameters were assayed and the correlations between these parameters and MDS clone burden were analysed.
RESULTSThe clonal burden of MDS patients was (67.4 +/- 36.2)%. It correlated positively with bone marrow blasts (r = 0.483, P < 0.05), negatively with hemoglobin level (r = -0.445, P < 0.05). The number of blasts, hemoglobin and erythrocytes in high clonal burden (>50%) and low clonal burden (< or = 50%) groups were significantly different (P < 0.05). CD4+ T lymphocytes of MDS patients and normal controls were (274.18 +/-71.85) x 10(6)/L and (454.82 +/- 205.88) x 10(6)/L (P < 0.05) respectively. CD8+ T lymphocytes between MDS patients and normal controls had no difference. The serum level of IL-2 of MDS patients and normal control groups were (6.29 +/- 3.58) g/L and (3.11 +/- 1.40) microg/L (P < 0.05) respectively; but no difference in the serum level of TNF between MDS and control groups. The ratio of CD4+ to CD8+ in high clonal burden patients was 1.90 + 0.52, and in low clonal burden patients was 0.97 +/- 0.44 (P < 0.05).
CONCLUSIONThe clonal burden and deficient T cell immunity are the indicators for predicting MDS patients clinical progression.
Adolescent ; Adult ; Aged ; Bone Marrow Cells ; pathology ; Chromosome Aberrations ; Female ; Humans ; Male ; Middle Aged ; Myelodysplastic Syndromes ; genetics ; immunology ; pathology ; T-Lymphocytes ; immunology
7.Prognostic value of Montreal Cognitive Assessment in heart failure patients.
Si Qi LYU ; Hui Qiong TAN ; Shao Shuai LIU ; Xiao Ning LIU ; Xiao GUO ; Dong Fang GAO ; Ran MO ; Jun ZHU ; Li Tian YU
Chinese Journal of Cardiology 2020;48(2):136-141
Objective: To explore the occurrence of cognitive impairment in Chinese heart failure (HF) patients and it's impact on prognosis. Methods: In this prospective observational study, a total of 990 HF patients were enrolled from 24 hospitals in China during December 2012 to November 2014. All patients were administrated with the interview-format Montreal Cognitive Assessment (MoCA), according to which they were divided into MoCA<26 (with cognitive impairment) group and MoCA≥26 (without cognitive impairment) group. Baseline data were collected and a 1-year follow up was carried out. Univariate and multivariate logistic or Cox regression were performed for 1-year outcomes. Results: Cognitive impairment was evidenced in 628 patients (63.4%) and they were more likely to be older, female, and with higher proportion of New York Heart Association(NYHA) class Ⅲ-Ⅳ, chronic obstructive pulmonary disease (COPD), ischemic heart disease, while body mass index (BMI), education level, and medical insurance rate were lower (all P<0.05) as compared to patients in MoCA≥26 group. The rate of percutaneous intervention, device implantation, cardiac surgery and evidence-based medications were significantly lower in MoCA<26 group than in MoCA≥26 group (all P<0.05). During the 1-year follow up, patients in the MoCA<26 group had higher all-cause mortality (10.2%(64/628) vs. 2.2%(8/362), P<0.01), cardiovascular mortality (5.9%(37/628) vs. 0.8%(3/362), P<0.01) and major adverse cardiac and cerebrovascular events (MACCE) (9.6%(60/628) vs. 2.5%(8/362), P<0.01) than patients in the MoCA≥26 group. In univariate regression, MoCA<26 was associated with increased all-cause mortality (HR(95%CI):4.739(2.272-9.885), P<0.01), cardiovascular mortality (HR(95%CI):7.258(2.237-23.548), P=0.001) and MACCE (OR(95%CI):4.143(2.031-8.453), P<0.01). After adjustment by multivariate regression, MoCA<26 was indicated as an independent risk factor for all-cause mortality (HR(95%CI): 6.387(2.533-16.104), P<0.01), cardiovascular mortality (HR(95%CI): 10.848(2.586-45.506), P=0.001) and MACCE (OR(95%CI): 4.081(1.299-12.816), P=0.016), while not for re-hospitalization for HF (OR(95%CI):1.010(0.700-1.457), P=0.957). Conclusions: Cognitive impairment is common in HF patients,and it is an independent prognostic factor for 1-year outcomes. Routine cognitive function assessment and active intervention are thus recommended for HF patients.
China
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Female
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Heart Failure
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Humans
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Mental Status and Dementia Tests
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Prognosis
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Prospective Studies
8.An analysis of relapse and risk factors of autoimmune hemolytic anemia and Evans syndrome.
Hong LIU ; Zong-hong SHAO ; Zhen-zhu CUI ; Yu-hong WU ; Tie-jun QIN ; Rong FU ; Guang-sheng HE ; Jun SHI ; Jie BAI ; Yan-ran CAO ; Chong-li YANG ; Tian-ying YANG
Chinese Journal of Hematology 2003;24(10):534-537
OBJECTIVETo analyse the relapse rate and risk factors of autoimmune hemolytic anemia (AIHA) and Evans syndrome.
METHODSFifty two cases of AIHA and Evans syndrome in remission being followed up for 1 - 14 years (median time 3.8 years) were analysed for relapse rate. The risk factors of relapse were analysed by case-control study.
RESULTSThe total relapse rate of these AIHA and Evans syndrome patients was 57.7%, and the median remission duration to the first relapse was 9 months. The relapse rates in patients with negative Coombs test, warm autoantibodies and both of warm and cold autoantibodies were 30.8% (4/13), 54.0% (13/24) and 86.7% (13/15), respectively. The relapse rate in patients with cold antibody was the highest (P < 0.05). The relapse rate in patients with antibody titer >or= 100 was 92.9% (13/14) and was higher than that in patients with antibody titer < 100 [59.5% (13/22)] (P < 0.05). Patients treated with prednisone and cyclosporin relapsed less than those treated with prednisone alone, and the relapse was related to the therapy course of prednisone and CsA.
CONCLUSIONBecause of the high relapse rate, AIHA and Evans syndrome should be treated according to the class of autoantibodies, and with longer course of prednisone and cyclosporin and prophylaxis of infection.
Adolescent ; Adult ; Aged ; Anemia, Hemolytic, Autoimmune ; etiology ; immunology ; Autoantibodies ; blood ; Child ; Cyclosporine ; therapeutic use ; Female ; Humans ; Male ; Middle Aged ; Prednisone ; therapeutic use ; Recurrence ; Risk Factors ; Syndrome ; Thrombocytopenia ; etiology ; immunology
9.The clinical features of severe aplastic anemia patients with complication of infection.
Yu-hong WU ; Zong-hong SHAO ; Hong LIU ; Zhen-zhu CUI ; Tie-jun QIN ; Rong FU ; Guang-sheng HE ; Jun SHI ; Jie BAI ; Yan-ran CAO ; Tian-ying YANG ; Chong-li YANG
Chinese Journal of Hematology 2003;24(10):530-533
OBJECTIVETo study the clinical features of severe aplastic anemia (SAA) patients with complication of infection.
METHODSA retrospective analysis of prevalence of infection occurring in 229 SAA patients, their bacterial spectrum, and the effect of GM-CSF or G-CSF on the infection were done.
RESULTThe prevalence of infection in SAA patients was 86.0%, among which 54.2% was infected with gram-positive organisms, 40.0% with gram-negative bacilli and 5.8% with fungal infections. Septicemia occurred mostly with E. coli and Pseudomonas infection. Patient's neutropenia was significantly related to the infection. The patients with neutrophil count less than 0.2 x 10(9)/L had more frequent and severe infection. Age, hemoglobin level, subtype of T lymphocytes and antithymocyte globulin therapy were not related to infection. Prophylaxis usage of floxacin could not reduce patient' gastrointestinal infection. The total mortality of SAA patients with infection was 23.1%. Pulmonary infection and septicemia increased mortality, and GM-CSF/G-CSF therapy reduce mortality.
CONCLUSIONSAA patients were at high risk of infection which was significantly associated with severe neutropenia. GM-CSF or G-CSF therapy exerts an assistant role to antibiotics in controlling the infections.
Adolescent ; Adult ; Aged ; Anemia, Aplastic ; complications ; Anti-Bacterial Agents ; therapeutic use ; Bacteria ; isolation & purification ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Infection ; drug therapy ; etiology ; Male ; Middle Aged ; Time Factors
10.Expression of apoptosis-related proteins in bone marrow neutrophils of patients with paroxysmal nocturnal hemoglobinuria.
Yan-Ran CAO ; Zong-Hong SHAO ; Hong LIU ; Ming-Feng ZHAO ; Guang-Sheng HE ; Jun SHI ; Jie BAI ; Rong FU ; Mei-Feng TU ; Hua-Quan WANG ; Li-Min XING ; Zheng-Zhu CUI ; Juan SUN ; Hai-Rong JIA ; Tian-Ying YANG
Journal of Experimental Hematology 2005;13(5):871-874
This study was aimed to evaluate expression levels of CD166, Fas and apoptosis-related proteins in bone marrow neutrophils of PNH patients and normal controls, and to analyze their correlation in order to explore whether exist apoptosis abnormality in BM neutrophils of PNH patients. The expression levels of CD16b, Fas and Bax, Bcl-2 in BM neutrophils of PNH patients and normal controls were assayed by flow cytometry; the difference of expression levels between patients and controls, and expression correlation between CD16b and apoptosis-related proteins were compared. The results showed that (1) the expression levels of CD16b on BM neutrophils of patients and controls were (20.36 +/- 9.05)% and (71.34 +/- 26.8)% respectively (P = 0.01); (2) the expression levels of CD95 on BM neutrophils of patients and controls were (62.83 +/- 32.11)% and (48.00 +/- 38.52)% respectively, there were no significant difference between CD95 expressions in BM neutrophils of PNH patients and controls and no significant correlation between expression of CD95 and CD16b on BM neutrophils of PNH patients (P > 0.05); (3) the expression levels of Bcl-2 in BM neutrophil cytoplasma of patients and controls were (8.64 +/- 5.40)% and (16.82 +/- 15.39)% respectively, there were no significant difference between Bcl-2 expression of patients and controls, and no significant correlation between the expression of Bcl-2 and CD16b in BM neutrophil cytoplasma of PNH patients (P > 0.05); (4) the expression levels of Bax in BM neutrophil cytoplasma of patients and control were (30.47 +/- 22.15)% and (48.47 +/- 15.99)% respectively, there were no significant difference between the Bax expressions of patients and controls, and no significant correlation between the Bax and CD16b expressions in BM neutrophil cytoplasma of PNH patients. In conclusion, BM neutrophils of PNH patients expressed apoptosis-related CD95, Bcl-2 and Bax without significant difference from the normal controls, and without significant correlation with the CD16b expression. It is suggested that the cell growth and decrease of PNH patients possibly are independent of abnormal apoptosis.
Adolescent
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Adult
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Apoptosis Regulatory Proteins
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biosynthesis
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Bone Marrow Cells
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metabolism
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pathology
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Female
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Flow Cytometry
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GPI-Linked Proteins
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Hemoglobinuria, Paroxysmal
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metabolism
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pathology
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Humans
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Male
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Middle Aged
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Neutrophils
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metabolism
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pathology
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Proto-Oncogene Proteins c-bcl-2
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biosynthesis
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Receptors, IgG
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biosynthesis
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bcl-2-Associated X Protein
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biosynthesis
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fas Receptor
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biosynthesis