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.

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.

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) vs. 8%(3/36), χ²=4.93, P=0.032], serum total bilirubin (TBIL) [17.3(12.2, 28.2) μmol/L vs. 14.5 (9.4, 21.1) μmol/L, Z=2.25, P=0.024], direct bilirubin (DBIL)[5. 5 (3.4, 12.4) μmol/L vs. 4.4(2.9, 7.1) μmol/L, Z=2.00, P=0.045], and glutamyl aminotransferase (GGT)[193.0(64.3, 454.8)U/L vs. 105.5(53.5, 179.5)U/L, Z=2.02, P=0.043], alkaline phosphatase(ALP)[183(86, 351)U/L vs. 135(85, 188) U/L, Z=1.98, P=0.048] in PBC patients with dyslipidemia were significantly higher than thosein patients with normal serum lipids.Pearson regression analysis showed that in PBC patients with dyslipidemia, the ALT was positively corrected with TG and TC(r=0.248 and 0.272, P=0.015 and 0.008); ALB was positively correlated with LDL-C(r=0.335, P=0.002); DBIL was positively corrected with HDL-C(r=0.252, P=0.022); TC was positively correlated with ALP and GGT(r=0.313 and 0.346, P=0.002 and 0.001); GGT was positively correlated with LDL-C(r=0.251, P=0.022). Conslusion Increased TC and LDL-C were more common in PBC patients. PBC patients with dyslipidemia have more severe liver damage than the patients with normal serum lipids.

Clinical data of 3 patients with linezolid-induced lactic acidosis treated in 2 hospitals in Beijing and Shandong were retrospectively analyzed,and relevant literatures at home and abroad were retrieved.The pathogene-sis,risk factors,clinical manifestations,and treatment scheme were valuated.Three patients received linezolid anti-infection treatment due to their disease condition.At different phases of treatment,they developed lactic acidosis that was difficult to be explained with septic shock.After discontinuing linezolid and receiving bedside continuous veno-venous hemofiltration(CVVH)treatment,patient's blood lactate levels decreased significantly.It is clinically diagnosed linezolid-induced lactic acidosis finally,but the initial diagnosis was septic shock by all doctors.After ac-tive treatment,2 patients recovered and 1 patient died.Linezolid-induced lactic acidosis is easily to be misdiagnosed as septic shock.Clinicians should enhance their understanding and recognition of the early diagnosis of the adverse reactions,take effective measures,so as to improve patient prognosis.

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

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

OBJECTIVETo study the characteristics of cell cycle and proliferation of CD34+ hematopoietic stem cells in patients with myelodysplastic syndromes (MDS).

METHODSPropidium iodide staining was used to examine cell cycle parameters (G(0)/G(1), S and G(2)/M) of bone marrow mononuclear cells (BMMNCs) while immunofluorescent double staining and FACS techniques were used to measure Ki67 expression in BM CD34+ cells from normal control, patients with MDS, acute myeloid leukemia preceded by MDS (MDS-AML) and primary AML.

RESULTSThere was a statistical up-tendency in G(0)/G(1) phase proportion of BMMNCs whereas a statistical down-tendency in S and G(2)/M phase proportions among normal control, MDS and primary AML. Compared to primary AML, MDS-AML had significantly higher ratios of S (P < 0.05), G(2)/M (P < 0.05) and S + G(2)/M (P < 0.05) phase cells while lower ratio of G(0)/G(1) phase cells (P < 0.05). The proportion of CD34+Ki67+ cells in MDS patients was significantly higher than that in normal control (P = 0.004). So were the percentages of CD34+Ki67+ cells in low-risk [(0.54 +/- 0.49)%, P < 0.05] and high-risk MDS patients [(1.69 +/- 1.66)%, P = 0.022]. Furthermore, there was statistical difference between low-risk and high-risk MDS (P < 0.05). Compared to normal control and primary AML, MDS-patients had the highest proportion of CD34+Ki67+ cells [(16.75 +/- 13.58)%, P < 0.05]. The proportion of CD34+Ki67+ cells in CD34+ cells in MDS patients [(48.50 +/- 20.49)%] was significantly higher than that in normal control [(27.71 +/- 16.04)%, P < 0.01]. So were the low-risk [(51.85 +/- 21.80)%, P = 0.002] and high-risk MDS [(43.93 +/- 18.57)%, P < 0.05]. The proportion of CD34+Ki67+ cells in CD34+ cells in MDS-AML patients [(60.92 +/- 30.12)%] was the highest, and was statistically higher than that in both normal control (P < 0.01) and primary AML patients [(17.01 +/- 15.93)%, P < 0.001]. The proportion of CD34+Ki67+ cells in Ki67+ cells in MDS patients [(4.91 +/- 4.68)%, P < 0.01] was significantly higher than that [(2.43 +/- 2.37)%] in normal controls. In the low-risk MDS group it was (4.11 +/- 3.94)%, (P > 0.05) and in high-risk MDS group it was (5.76 +/- 5.38)%, (P < 0.05).

CONCLUSIONHigh proportion of G(0)/G(1) cells and G(1) phase arrest occurred in MDS. High proliferation capacity of MDS clone, especially that derived from CD34+ cells, might play an important role in the clonal expansion, diseases deterioration and worse prognosis of MDS.

Acute Disease ; Adolescent ; Adult ; Aged ; Antigens, CD34 ; blood ; Bone Marrow Cells ; metabolism ; pathology ; Cell Cycle ; Cell Proliferation ; Female ; Flow Cytometry ; Fluorescent Antibody Technique ; Humans ; Ki-67 Antigen ; blood ; Leukemia, Myeloid ; blood ; Male ; Middle Aged ; Myelodysplastic Syndromes ; blood ; Young Adult

OBJECTIVETo investigate the role of the burden of abnormal hematopoietic clone in the development of myelodysplastic syndromes (MDS).

METHODSThe ratio of the bone marrow cells with abnormal chromosomes to the total counted bone marrow cells was regarded as the index of MDS clone burden. The disease severity related parameters including white blood cell count, hemoglobin, platelet count, lactate dehydrogenase level, bone marrow blast, myeloid differentiation index, micromegakaryocyte, transfusion, interleukin-2, tumor necrosis factor (TNF), CD4+ and CD8+ T cells of MDS patients were assayed, and the correlations between those parameters and MDS clone burden were also analyzed.

RESULTSThe clone burden of MDS patients was 67.4% +/- 36.2%. MDS clone burden positively correlated 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 clone burden (> 50%) and low clone burden ( < or = 50%) groups were 7.78% +/- 5.51% and 3.45% +/- 3.34%, 56.06 +/- 14.28 g/L and 76.40 +/- 24.44 g/L, (1.82 +/- 0.48) x 10(12)/L and (2.32 +/- 0.66) x 10(12)/L, respectively (all P < 0.05). CD4+ T lymphocytes of MDS patients and normal controls were (0.274 +/- 0.719) x 10(9)/L and (0.455 +/- 0.206) x 10(9)/L, respectively (P < 0.05). CD8+ T lymphocytes of MDS patients and normal controls were (0.240 +/- 0.150) x 10(9)/L and (0.305 +/- 0.145) x 10(9)/L, respectively. The serum level of interleukin-2 of MDS patients (6.29 +/- 3.58 ng/mL) was significantly higher than normal control (3.11 +/- 1.40 ng/mL, P < 0.05). The serum level of TNF of MDS patients and normal control group were 2.42 +/- 1.79 ng/mL and 1.68 +/- 0.69 ng/mL, respectively. The ratio of CD4 to CD8 was higher in high clone burden MDS patients (1.90 +/- 0.52) than that in low clone burden patients (0.97 +/- 0.44, P < 0.05).

CONCLUSIONThe quantitive clonal karyotype abnormalities and deficient T cell immunity are important parameters for evaluating MDS severity and predicting its progression.

Adolescent ; Adult ; Aged ; Bone Marrow Cells ; pathology ; Case-Control Studies ; Chromosome Aberrations ; Female ; Hematopoiesis ; genetics ; Hematopoietic Stem Cells ; pathology ; Humans ; Male ; Middle Aged ; Myelodysplastic Syndromes ; blood ; genetics ; pathology ; Neoplastic Stem Cells ; pathology ; Polycythemia ; genetics ; pathology ; T-Lymphocyte Subsets ; pathology ; Young Adult

OBJECTIVETo investigate the prognostic value of quantitative chromosomal abnormality in myelodysplastic syndromes (MDS).

METHODSChromosomal karyotypes in seventy-one MDS patients' were analyzed quantitatively. Based on the number of abnormal metaphase in 20 counted metaphases, the patients were divided into three groups: no abnormal karyotypes, abnormal metaphases less than or equal to five, and that more than five. The leukemia transformation rate, death rate and survival time between these three groups were compared.

RESULTSForty-four cases (62.0%) had abnormal karyotypes. The incidences of abnormal karyotypes in RA, RCMD and RAEB were 76.9%, 55.8% and 75.0%, respectively, being no significant difference (P > 0.05). Among the abnormal karyotypes, complex abnormality with two or more abnormal karyotypes was most common and accounted for 47.7%. The frequencies of trisomy 8, monosomy 7 and del 20q were 18.2%, 4.5% and 4.5%, respectively. Other kinds of abnormal karyotypes totally accounted for 25%. There were 27 cases of group 1, 28 of group 2 and 16 of group 3. Eighteen cases (25.4%) transformed to acute leukemia. The incidences of leukemia transformation in group 1, 2 and 3 were 18.5%, 25% and 37.5%, and the death rates were 29.6%, 42.9% and 56.3%, respectively. The median survival times were 60, 47 and 24 months respectively.

CONCLUSIONThe quantitative chromosome abnormality has prognostic value in MDS.

Adolescent ; Adult ; Aged ; Chromosome Aberrations ; Female ; Follow-Up Studies ; Humans ; Karyotyping ; Male ; Middle Aged ; Myelodysplastic Syndromes ; genetics ; Prognosis

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