Role of sweet taste receptors in the activation of NLRP3 inflammasome signaling in diabetic kidney disease / 中华内分泌代谢杂志

Objective@#To investigate the role of sweet taste receptors(STRs)in the activation of reactive oxygen species (ROS)-NLRP3 inflammasome signaling in diabetic kidney disease(DKD).@*Methods@#DKD mouse were established by high-fat diet and streptozotocin. After mouse glomerular mesangial cells(GMCs)were exposed to high glucose, STRs(T1R2/T1R3)and associated signaling components and NLRP3 inflammasome signaling components expressions were detected. After GMCs were treated with STRs inhibitor lactisole, the production of ROS was detected and the role of STRs in the activation of NLRP3 signaling was investigated.@*Results@#Under high glucose condition, the expressions of T1R2, T1R3, Gα-gustducin, phospholipase C-β2, and TRPM5 were significantly decreased in vivo and in vitro(all P<0.05)and ROS-NLRP3 signaling was activated(all P<0.05). Lactisole significantly mitigated the production of ROS and the activation of NLRP3 inflammasome signaling stimulated by high glucose in GMCs(all P<0.05).@*Conclusion@#The STRs(T1R2/T1R3)-mediated signaling pathway may be involved in the regulation of ROS-NLRP3 inflammatory signaling, suggesting that STRs may act as new therapeutic targets of DKD. (Chin J Endocrinol Metab, 2018, 34: 587-593)

Effects of short-chain fatty acids on oxidative stress and inflammation in glomerular mesangial cells induced by high glucose / 中华内分泌代谢杂志

To investigate the effects of short-chain fatty acids(SCFAs)on oxidative stress and inflammation in cultured glomerular mesangial cells(GMCs)under high glucose.SV-40 MES 13 mouse GMCs were exposed to 30 mmol/L glucose, exogenous SCFAs or their specific G-protein coupled receptor 43(GPR43)agonist were used individually as an intervention.GPR43 expression was suppressed by transfection with GPR43-specifc siRNA.The oxidative stress-related factors reactive oxygen species and malondialdehyde were detected and the expression of GPR43,monocyte chemotactic protein 1(MCP-1),and interleukin-1β(IL-1β)were observed.GPR43 expression were significantly down-regulated,but reactive oxygen species and malondialdehyde production and MCP-1 and IL-1β releases were induced by high glucose(all P<0.05),treatment with SCFAs or GPR43 agonist reversed high glucose-induced oxidative stress and inflammation in a dose-dependent manner(all P<0.05).However, these SCFAs-mediated effects were blunted by siRNA-mediated knockdown GPR43(all P<0.05).SCFAs mitigates high glucose-induced oxidative stress and inflammatory injury in part via GPR43 signaling pathway,suggesting a likely mechanism for SCFAs-induced therapeutic effect in diabetic kidney disease.

Clinical analysis of 144 cases of infant leukaemia / 南方医科大学学报

<p><b>OBJECTIVE</b>To explore the clinical features, laboratory findings and treatment of infant leukemia.</p><p><b>METHODS</b>A retrospective analysis of the clinical data was performed of the cases with the diagnosis of infant acute leukemia from August 1993 to October 2014 in our hospital.</p><p><b>RESULTS</b>A total of 144 cases of infant leukemia were diagnosed in the defined period, including 83 cases of acute lymphoblastic leukemia, 55 myeloid leukemia, 1 hybrid acute leukaemia and 5 with incompatible cytological and immunophenotyping findings. The patients at the age of 9 to 12 months accounted for the largest proportion (38.2%), and 87.5% of the patients had hepatosplenomegaly; Six patients below 6 months old had skin infiltration. In about 1/3 of the patients, the white blood cells count was no greater than 100 × 10⁹ /L. Ninety-five patients had chromosome examinations, which identified chromosome abnormalities in 67 patients, including 18 positive for t(4;11)or t(9;11)or t(11;19), and younger patients were more likely to have chromosome abnormalities. Thirty-seven patients underwent MLL gene detection and 11 of them had positive results; the positive patients had higher rate of chromosome 11 abnormalities than the negative patients. Most of the patients gave up treatments after diagnosis and only 6 patients older than 6 months completed regular chemotherapeutic treatments and were now in complete remission.</p><p><b>CONCLUSION</b>Infant leukemia is a rare type of leukemia with different clinical features from other types of leukemia. The patients often present with hepatosplenomegaly, high white blood cell counts, MLL gene fusion, and chromosome 11 abnormalities. The prognosis of infant leukemia is not favorable, and the current treatment still relies on chemotherapy.</p>

Glycogen synthase kinase 3β and leukemia / 国际肿瘤学杂志

Glycogen synthase kinase 3β (GSK3 β) is a conserved cytoplasmic serine/threonine protein kinase.It can not only negatively regulate the classical Wnt and Akt signal pathways which are associated with leukemia,but also positively regulate the NF-κB,non-classical Wnt and lysosomal apoptosis signal pathways,thereby regulating the proliferation,differentiation and apoptosis of leukemia cells.At present,a variety of GSK.3 β inhibitors and activators have been reported in the researches of leukemia,which play important roles in reducing the adverse reactions of chemotherapy,reversing multidrug resistance and enhancing killing effect to leukemia cells.GSK3β is expected to become a target of leukemia molecular targeted therapy.

Molecular mechanism of tetramethylpyrazine to induce human promyelocytic HL-60 leukemia cells differentiation / 中国中药杂志

<p><b>OBJECTIVE</b>To study the molecular mechanism of tetramethylpyrazine to induce human promyelocytic HL-60 leukemia cells differentiation.</p><p><b>METHOD</b>The cell proliferation was determined by MTT. The differentiation of the cells was detected by NBT reduction test. Cellular morphology was observed by Wright's staining. Cell cycle distribution and the distribution of CD11b, CD14 were detected by flow cytometry. Then RT-PCR and Western blot assay were employed to detect the expressions of c-myc, p27, CDK2 and cyclinE1 in HL-60 cells after exposure to TMP.</p><p><b>RESULT</b>TMP inhibited the proliferation in a dose and time dependent manner. TMP at the concentration of 200 mg x L(-1) to 300 mg x L(-1) induced unterminal differentiation of HL-60 cell and synergistically blocked the cell cycle progression of HL-60 cells in G0/G1 phase. The expression of c-myc was down-regulated as well as the protein expression of cyclin E and CDK2, while the mRNA and protein expression of P27 were remarkably up-regulated.</p><p><b>CONCLUSION</b>Small doses of TMP induces differentiation of HL-60 cells throughout the cell cyde, as detected by a slower rate of accumulation in G0/G1, possibly by regulating the expression and activity of G1/S phase-related molecules.</p>

Influence of VLA-4 antibody combined with VP_(16) upon childhood leukemic cells' apoptosis / 白血病·淋巴瘤

Objective To investigate the influence of very late antigen-4 (VLA-4) antibody and VLA-4 antibody combined with VP_(16) in vitro on childhood leukemic cells' apoptosis and explore the protection of bone marrow stromal cells (BMSC) upon leukemic cells and its related mechanisms. Methods Leukemic bone marrow stromal cells were isolated by human lymphocyte separation medium and in vitro culture of BMSC (adherent) and leukemia cells (suspended) BMSC+leukemic cells group were as control. Then VLA-4 antibody and/or VP_(16) were added respectively to VLA-4 antibody group, VP_(16) group and VLA-4 antibody combined with VP_(16) group to detect the apoptosis of leukemic cells in different groups through Annexin V-FITC double-labeled flow cytometry and the expression of Survivin, bcl-2 genes in each group of leukemic cells detected by RT-PCR. Results The results showed by flow cytometry that compared with the control groups, for 12 h or 24 h, the early and total apoptosis rates of leukemic cells of the three experimental groups were significantly increased(P <0.05); the early and total apoptosis rates of leukemic cells treated with VLA-4 antibody combined with VP_(16) group was markedly increased, compared with the control group (P <0.05); the comparison of the early and total apoptosis rates for the three experimental groups between 12 h and 24 h was significantly different (P<0.01). Moreover, RT-PCR results showed that the expression of Survivin and bcl-2 genes of leukemic cells in three experimental groups was reduced in varying degrees and the reduction of VLA-4 antibody combined with VP_(16) group was the most obvious. Conclusion BMSC plays a protective role on leukemic cells, and VLA-4 antibody can block the adhesion between BMSC and leukemic cells promoting leukemic cells apoptosis and enhance the sensibility of apoptosis of leukemic cells induced by chemotherapeutics.

In silicon cloning of HV126, a novel human gene related to multi drug resistance in leukemia / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To find the novel gene related to the multi-drug resistance in leukemia and explore the molecular mechanism of multi-drug resistance.</p><p><b>METHODS</b>The subtracted HL-60/VCR cDNA library was generated through the suppression subtractive hybridization using the wild HL-60 cells' cDNA as target and HL-60/ ATRA cells' as driver. A novel expression sequence tag (EST) sequence, which differentially expressed in HL-60/ ATRA cell, was screened by cDNA chip. Then a novel human gene, HV126 was assembled by the EST assembly tools. Bioinformatical databases and softwares were used to analyze and predict its function. Reverse transcription-PCR (RT-PCR) was used to detect the expression of HV-126 gene in leukemia cells before and after chemotherapy.</p><p><b>RESULTS</b>The full open reading frames (ORFs) of the novel EST assembled by overlapping dbEST sequences included a 1991 bp nucleic sequence, which was named HV126. The deduced amino acid sequence consisted of 365 amino acids. The sequence of the novel gene exhibited 43% homology to a known gene, which is a possible member of the death domain-flood family implicated in apoptosis and inflammation. The expression of HV126 was proved to be related to the drug sensitivity in leukemia cells by RT-PCR.</p><p><b>CONCLUSION</b>HV126, the novel gene, might have roles in regulating multi-drug resistance in leukemia. Further laboratory research should be done on cloning and making clear the gene function.</p>

Effect of enhanced survivin expression on lymphocyte proliferation and function in K562 cells / 第三军医大学学报

Objective To study the effect of survivin gene expression on the lymphocyte’s proliferation and function in cultured K562 cells. Methods The constructed recombinant vector pEGFP-C1-survivin and the plasmid pTZU6+1-survivin encoding short hairpin RNA of survivin were transfected into K562 cells respectively to generate K562/survivin+ cells and K562/survivin-cells. K562/survivin+ cells were selected by G418. The survivin mRNA and protein levels in the 3 kinds of cells (K562/survivin+,K562 and K562/survivin-) were detected by semi-quantitative RT-PCR and immunohistochemistry methods. The peripheral blood mononuclear cells (PBMC) from healthy subjects were co-cultured with these 3 cells respectively in mixed lymphocyte-tumor cell culture (MLTC). Lymphocyte proliferation in the supernatant was evaluated by MTT assay. Nature killer activity was detected by flow cytometry and IFN-? level was measured by ELISA assay. Results Proliferation index in K562/survivin+ group was much lower than that in the other 2 groups (P

Expression and clinical value of surviving in mononuclear cells of bone marrow and peripheral blood of children with acute leukemia / 第三军医大学学报

Objective To study the survivin expression in mononuclear cells of both bone marrow and peripheral blood in childhood acute leukemia(AL).Methods Twenty-one children with AL were divided into newly diagnosed group(n=12) and complete remission(CR) group(n=9).The control group was consisted of 15 children without tumor.Mononuclear cells were separated from bone marrow and peripheral blood (PBL) by gradient density centrifugation and RNA was extracted,and RT-PCR was performed to detect survivin mRNA.Results In newly diagnosed AL group,6 out of 12 patients had positive survivin expression in bone marrow(50%) and 4 out of 11 patients in the PBL(36.37%),both of which were higher than the control group(P0.05).In 9 CR patients,all the patients had the negative expression in the PBL,but 1 positive expression in bone marrow(11.11%).The positive expression in CR group was significantly lower than the newly diagnosed AL group.The only one patient with positive survivin expression in CR group had relapse 7 weeks later.In newly diagnosed patients,the average immature leukocytes were(76.43?43.28)?10~(9)/L in the patients with positive survivin expression and(13.85?5.86)?10~(9)/L in those with negative survivin expression(P

Construction of recombinant adenovirus vector encoding VEGF siRNA and its inhibitory effect on K562 cell proliferation / 第三军医大学学报

Objective To construct an recombinant adenovirus vector of vascular endothelial growth factor small interfering RNA(Ad5-VEGFsi) and to observe its inhibitory effect on the cell proliferation of human leukemia K562 cells. Methods The specific human VEGF siRNA was subcloned into the shuttle plasmid pSES-HUS and then cotransfected with plasmid pAdeasy-1 to produce pAd5-VEGFsi by homologous recombination. The identified recombinant plasmid pAd5-VEGFsi was packaged and amplified in 293 cells. At 72 h after the transfection of Ad5-VEGFsi into K562 cells, VEGF mRNA expression and the protein level of VEGF in the cell culture supernatant were determined by RT-PCR and ELISA, respectively. Cell proliferation was measured by MTT assay and cell apoptosis by flow cytometry. Results The recombinant adenovirus Ad5-VEGFsi was successfully constructed. Ad5-VEGFsi with the titer of 4.6?1011 pfu/ml was harvested by CsCl gradient purification. Compared with those in the control, EGF mRNA in cells decreased by 66.55% (P

Expressions of Livin ? and Livin ? in marrow mononuclear cells of childhood acute leukemia and the clinical significance / 第三军医大学学报

Objective To investigate the expressions of Livin ? and Livin ? in marrow mononuclear cells(MMNCs)of childhood acute leukemia(CAL)and explore the clinical significance.Methods Real-time quantitative PCR and Western blot were used to detect the mRNA and protein expressions of Livin ? and Livin ? in MMNCs of CAL,respectively.Results Both Livin? and Livin? expression rates and expression levels were higher in preliminary diagnosis group(n=51)of CAL,including acute lymphoblastic leukemia(n=39)and acute myeloid leukemia(n=12),compared with the control group(P5%)to induction chemotherapy of ALL than in the patients without chemotherapy(P

To Promote the Course Construction on the Basis of the Subject Development / 中华医学教育探索杂志

The internal medicine of paediatrics is the important major course in paediatrics. Focusing on strengthening the constructionof staff, we cultivate all levels of teachers from different administrative level and personality, do schorlarly research meticulously,standardizing management and improve the teaching evaluation system. Besides we promote the construction and development of theinternal medicine of paediatrics by carrying out teaching research, impsoving teaching methods and making paediatics the key subjectin China.

Forging and training of interns’clinical thinking ability / 中华医学教育探索杂志

The clinical thoughts of interns have some features such as insufficiency of initiative and enthusiasm and the absence of systemic. It is necessary to develop their good habits of active and diligent thinking as well as comprehensive thinking ability. Clinical teaching rounds, small lectures,discussion of complicated cases and interactive teaching are the specific measures for our medical interns to improve their ability of clinical thoughts effectifally.