Decitabine Enhances the Sensitivity of Leukemia Stem Cell to Allo-NK Cell-Mediated Killing / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the allo-NK cell-mediated killing effect enhanced by decitabine on leukemia stem cells(LSC) and the underlying mechanisms.</p><p><b>METHODS</b>LSC were separated from KG1a cells by using immunomagnetic beads. Allo-NK cells were isolated and purified from PBMC of healthy donors. Cytotoxicity of allo-NK cells against LSC were measured by LDH releasing assay. The apoptosis induced by allo-NK cells in LSC and the expressions of NKG2D ligands including MICA/B and ULBP1-3 on LSC were detected by flow cytometry.</p><p><b>RESULTS</b>The killing rate of allo-NK cells to LSC treated with 10 µmol/L decitabine for 24 hours was significant higher than that to LSC without treatment(60.52%±3.52% vs 22.08%±2.07%, 73.93%±2.33% vs 28. 99%±3.13%, 83.08%±1.32% vs 36.44%±2.40%, respectively)at the effector-target ratios of 5:1, 10:1, 20:1 (P<0.05). At the effector-target ratio of 10:1, decitabine significantly enhanced the apoptosis of LSC induced by allo-NK cells (7.84%±0.34% vs 3.33%±0.64%)(P<0.05). The expressions of NKG2D ligands(MICA/B,ULBP1,ULBP2,ULBP3) on LSC treated with decitabine 10 µmol/L for 24 hours were significantly increased (P<0.05).</p><p><b>CONCLUSION</b>Decitabine may enhance the allo-NK cell-mediated killing effects on LSC by up-regulation of the expressions of NKG2D ligands on LSC.</p>

Association of osteopontin gene polymorphism with lupus nephritis in Chinese Han population / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the possible association of a single nucleotide polymorphism (SNP) at position 9250 in exon 7 of osteopontin gene (OPN gene 9250) with lupus nephritis (LN) in southern Chinese patients.</p><p><b>METHODS</b>Altogether 109 patients with LN (12 males and 97 females) diagnosed according to the American College of Rheumatology criteria (revised in 1982) for systemic lupus erythematosus (SLE) and 180 healthy ethnically matched controls (34 males and 146 females), all Han people living in South China, were enrolled in this study with informed consent. Blood samples from all subjects were collected in EDTA tube for genomic DNA extraction according to the standard isolation procedures. OPN gene 9250 polymorphism was typed by PCR-restriction fragment length polymorphism (PCR-RFLP).</p><p><b>RESULTS</b>All the genotype frequencies of OPN gene 9250 were in Hardy-Weinberg equilibrium. Compared with the controls, LN patients showed significantly lower frequency of TT genotype of OPN gene 9250 (70% vs 51%, P<0.05) and significantly higher frequency of TC genotype (29.4% vs 45%, P<0.05). There were significant differences in OPN gene 9250 allele and phenotype frequencies between LN patients and the controls (P<0.05). Compared with the female controls, the frequency of TT genotype of OPN gene 9250 was significantly lower in female LN patients (68.5% vs 54%, P<0.05), and there was a significant difference in OPN gene 9250 allele frequency between them (P<0.05).</p><p><b>CONCLUSION</b>OPN gene 9250 polymorphism appears to be associated with the susceptibility to LN in southern Chinese Han population.</p>

Clinical and pathological characteristics of patients with glomerular hematuria as the major manifestation / 南方医科大学学报

<p><b>OBJECTIVE</b>To compare the clinical and pathological characteristics between patients with and without glomerular hematuria.</p><p><b>METHODS</b>Totally 310 patients with isolated microscopic hematuria and 24-hour urinary protein <0.5 g were enrolled in this study, who were free of renal calculi, infections, or tumors. These patients were divided into glomerular hematuria group and non-glomerular hematuria group according examination by phase-contrast microscope, and their clinicopathological data were retrospectively analyzed.</p><p><b>RESULTS</b>Of the 209 patients identified to have glomerular hematuria, 46.41% had IgA nephropathy, 22.49% had small glomerular lesions, and 14.35% had non-IgA mesangioproliferative glomerulonephritis. In the patients with non-glomerular hematuria, 65.34% had IgA nephropathy, 21.78% had non-IgA mesangioproliferative glomerulonephritis, and 8.91% had small glomerular lesions. No significant differences were found in the patients' gender, age, course of disease or hypertension between the two groups (P>0.05), but renal insufficiency was more common in patients with glomerular hematuria.</p><p><b>CONCLUSION</b>IgA nephropathy is still the most frequent pathological finding in patients with hematuria, and even in non-glomerular hematuria patients severe renal pathological changes can be common.</p>

Involvement of mitochondria apoptotic pathway in the manumycin inducing apoptosis of U937 and HL-60 / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the apoptosis induced by manumycin in U937 and HL-60 cell lines, and to explore the role of mitochondria apoptotic pathway in manumycin-inducing apoptosis.</p><p><b>METHODS</b>Leukemic cells line U937 and HL-60 were treated by manumycin at 2 micromol/L for different time. Apoptosis of leukemia cells was detected by flow cytometry. The cytosolic proteins were extracted using a digitonin buffer. The protein expression of cytochrome C, caspase-9, caspase-8, and caspase-3 were determined by western blot. Mitochondrial membrane potential was detected by JC-1.</p><p><b>RESULTS</b>In U937 and HL-60 cells, manumycin induced mitochondrial depolarization after 6 h treatment. The average red/green fluorescence ratios at 6 h were significantly (P < 0.01) lower than those at time 0, being 0.51 +/- 0.07 and 0.41 +/- 0.06 for control group respectively. Manumycin induced cytochrome C release from the mitochondria into the cytosol after 6 h treatment, and activated caspase-9, caspase-8, and caspase-3 after a 16h treatment. The broad-spectrum caspase-inhibitor Z-VAD-fmk at 50 micromol/L was able to inhibit caspase cleavage completely, but only reduced the manumycin-induced apoptosis rates by 51.69% and 56.47% in U937 and HL-60, respectively.</p><p><b>CONCLUSION</b>Manumycin induced apoptosis in U937 and HL-60 cell lines via mitochondria apoptotic pathway.</p>

Osteopontin gene polymorphism in association with systemic lupus erythematosus in Chinese patients / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Osteopontin (OPN) is one kind of cytokine which can play a number of roles in promoting activation of T lymphocyte, regulating balance between Th1 and Th2, participating in cell-induced immunologic response and stimulating B lymphocyte to express multi-clone antibodies. Some researches have showed that OPN may be involved in the pathogenesis of systemic lupus erythematosus (SLE). The aim of this study was to investigate possible association of a single nucleotide polymorphism (SNP) at position 9250 in exon 7 of the OPN gene (OPN gene 9250) with SLE in Chinese patients.</p><p><b>METHODS</b>Totally 158 patients (18 males and 140 females) fulfilled the revised criteria for SLE by the American College of Rheumatology in 1982 and 180 healthy volunteer controls (34 males and 146 females), all from the south of China, consented to participate in the study. OPN gene 9250 polymorphism was detected by polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP).</p><p><b>RESULTS</b>The frequency of TT genotype of the OPN gene 9250 was significantly lower (52.5% vs 70%, P < 0.05) and the frequency of TC genotype of the OPN gene 9250 was significantly higher (43.7% vs 29.4%, P < 0.05) in SLE patients than in controls. There were significant differences in OPN gene 9250 allele and phenotype frequencies between the SLE patients and controls (P < 0.05). When the SLE patients and controls were separated into men and women, significant differences of frequencies were noted in TT genotype, TC genotype and allele of the OPN gene 9250 in women (P < 0.05) but not in men (P > 0.05).</p><p><b>CONCLUSIONS</b>OPN gene 9250 polymorphism appears to be associated with susceptibility to SLE in Chinese Han ethnic population.</p>