简介:摘要目的比较IgA肾病患者不同病理分级的血清IgA、C3及IgA/C3水平;探讨血清IgA、C3及IgA/C3水平在IgA肾病病理分级评估中的意义。方法选择我院320例经肾穿刺活检病理检查确诊为原发性IgA肾病患者,用散射比浊法检测患者血清IgA、C3水平及按Lee氏分级标准评估IgA肾病病理分级。结果Lee分级为Ⅰ、Ⅱ级与Lee分级为Ⅲ、Ⅳ、Ⅴ级患者IgA水平分别为(2.75±1.10)vs(2.85±1.26)g/L,(P﹥0.05);血清C3水平分别为(1.18±0.33)vs(1.04±0.29)g/L,(P﹤0.05);血清IgA/C3率分别为(2.71±1.19)vs(3.24±1.69)g/L,(P﹤0.05)。结论IgA/C3比值在评估IgA肾病病理分级有重要参考意义。
简介:摘要目的探讨病毒性心肌炎患者血清半乳糖凝集素-3(galectin-3,Gal-3)、正五聚体蛋白-3(pentraxin-3,PTX-3)表达水平及其诊断及预后评估价值。方法选取2018年1月—2019年7月本院收治的122例病毒性心肌炎患者为研究对象(心肌炎组),同期选取118例健康体检者作为对照组。采用酶联免疫吸附法检测血清Gal-3、PTX-3及心肌指标脑钠肽(brain natriuretic peptide,BNP)、肌酸激酶同工酶(creatine kinase MB,CK-MB)、心肌肌钙蛋白I(cardiac troponin I,cTnI)水平;采用Pearson法分析病毒性心肌炎患者血清Gal-3、PTX-3与心肌指标的相关性;采用受试者工作特征曲线(receiver operator charateristic curve,ROC曲线)分析血清Gal-3、PTX-3水平对病毒性心肌炎的诊断价值;采用多因素Logistic回归分析影响病毒性心肌炎患者预后的因素。结果心肌炎组血清Gal-3、PTX-3、BNP、CK-MB、cTnI水平高于对照组,差异有统计学意义(P>0.05)。病毒性心肌炎患者血清Gal-3与PTX-3呈正相关(r=0.594、P<0.05),且二者与BNP、CK-MB、cTnI均呈正相关(P<0.05)。血清Gal-3、PTX-3水平诊断病毒性心肌炎的曲线下面积(area under curve,AUC)分别为0.873、0.916,截断值分别为5.431 ng/mL、4.159 ng/mL,特异性分别为74.5%、84.7%,敏感度分别为84.4%、82.0%;二者联合诊断的AUC为0.969,特异性为95.9%,敏感度为88.5%。预后不良组病毒性心肌炎患者血清Gal-3、PTX-3水平高于预后良好组,差异有统计学意义(P<0.05)。Gal-3、PTX-3、cTnI是影响病毒性心肌炎患者预后不良的独立危险因素(P<0.05)。结论病毒性心肌炎患者血清Gal-3、PTX-3表达水平升高,二者对病毒性心肌炎诊断及预后评价均可能有重要价值。
简介:摘要目的探讨地塞米松(Dex)对成骨细胞增殖和凋亡的影响及潜在的作用机制。方法将人成骨细胞hFOB 1.19分为4组:对照组(不加药物),Dex组(300 μM Dex处理48 h),Dex+AA组(300 μM Dex+2 000 μM AA处理48 h)和Dex+NAC组(300 μM Dex+500 μM NAC处理48 h)。抗坏血酸(AA)和N-乙酰-L-半胱氨酸(NAC)均为活性氧(ROS)的清除剂。检测各组细胞的增殖率、凋亡率、ROS和碱性磷酸酶(ALP)水平以及PI3K/AKT/GSK3β信号通路的活性。结果Dex的半抑制浓度(IC50)约在300 μM。与对照组对比,Dex组细胞的细胞增殖率[(100.00±1.76)%比(51.47±5.62)%]和ALP活性[(100.00±0.83)%比(69.71±7.53)%]显著降低(均P<0.05),细胞凋亡率[(3.71±1.16)%比(18.42±3.19)%]和ROS水平[(1.49±0.37)比(4.40±1.08)]显著增加(均P<0.05)。与Dex组对比,Dex+AA组和Dex+NAC组细胞的细胞增殖率[(51.47±5.62)%比(89.32±6.74)%、(51.47±5.62)%比(94.58±5.01)%]和ALP活性[(69.71±7.53)%比(85.49±8.17%)、(69.71±7.53)%比(92.55±8.01)%]显著增加(均P<0.05),细胞凋亡率[(18.42±3.19)%比(9.68±2.23)%、(18.42±3.19)%比(8.97±2.07)%]和ROS水平[(4.40±1.08)比(2.05±0.61)、(4.40±1.08)比(1.96±0.43)]均显著降低(均P<0.05)。与对照组对比,Dex组细胞的p-PI3K[(0.98±0.17)比(0.26±0.08)]和p-AKT相对表达量[(0.56±0.10)比(0.19±0.06)]、p-PI3K/PI3K[(1.72±0.35)比(0.43±0.10)]和p-AKT/AKT比值[(0.66±0.19)比(0.21±0.08)]均显著降低(均P<0.05),p-GSK3β相对表达量[(0.12±0.04)比(0.49±0.09)]和p-GSK3β/GSK3β比值[(0.67±0.15)比(1.32±0.24)]均显著增加(均P<0.05)。与Dex组对比,Dex+AA组和Dex+NAC组细胞的p-PI3K[(0.26±0.08)比(0.76±0.12)、(0.26±0.08)比(0.80±0.12)]和p-AKT相对表达量[(0.19±0.06)比(0.38±0.11)、(0.19±0.06)比(0.42±0.08)]、p-PI3K/PI3K[(0.43±0.10)比(1.21±0.27)、(0.43±0.10)比(1.35±0.21)]和p-AKT/AKT比值[(0.21±0.08)比(0.45±0.04)、(0.21±0.08)比(0.45±0.07)]均显著增加(均P<0.05),p-GSK3β相对表达量[(0.49±0.09)比(0.26±0.05)、(0.49±0.09)比(0.22±0.07)]和p-GSK3β/GSK3β比值[(1.32±0.24)比(0.83±0.19)、(1.32±0.24)比(0.73±0.16)]均显著降低(均P<0.05)。结论Dex可通过ROS-PI3K/AKT/GSK3β信号通路诱导成骨细胞凋亡。
简介:AbstractBackground:Pharmacological factors used to induce insulin resistance (IR) in in vitro models may not mimic the full in vivo features of type 2 diabetes mellitus (T2DM). This study aimed to examine the ability of diabetic serum (DS) to induce IR and investigate whether adipose-derived mesenchymal stem cell conditioned medium (ADMSC-CM) reverses DS-induced IR.Methods:DS was obtained from newly diagnosed T2DM patients. IR was induced in differentiated 3T3-L1 cells by employing dexamethasone, tumor necrosis factor alpha (TNF-α), palmitate and DS. Glucose uptake (2-[N-[7-nitrobenz-2-oxa-1,3-diazol-4-yl] amino]-2-deoxyglucose(2-NBDG) uptake assay), intracellular levels of reactive oxygen species (ROS), and superoxide radicals (O2-) (fluorescence microscopy and fluorometry) were analyzed in control and experimental samples. mRNA expression of key genes involved in glucose transport and inflammation were analyzed by using reverse transcription polymerase chain reaction (RT-PCR). Pro-inflammatory cytokines and phospho-insulin receptor substrate (IRS) (Ser-307) protein expression were analyzed by fluorescence activated cell sorter analysis. Statistical significance was determined by using one-way ANOVA followed by Tukey's multiple comparison tests.Results:ADMSC-CM significantly increased the DS-mediated decrease in 2-NBDG uptake (11.01 ± 0.50 vs. 7.20 ± 0.30, P < 0.01) and reduced DS-driven ROS (fluorescence count, 6.35 ± 0.46 vs. 9.80 ± 0.10, P < 0.01) and O2- (fluorescence count, 3.00 ± 0.10 vs. 4.60 ± 0.09, P < 0.01) production. Further, the ADMSC-CM restored DS-induced down regulation GLUT4 (1.52- fold, P < 0.05) as well as the up-regulation of PPARγ (0.35-fold, P < 0.01), and IKKβ (0.37-fold, P < 0.01) mRNA, and phospho-IRS (Ser-307) protein expression compared to the baseline (median fluorescence intensity, 88,192 ± 2720 vs. 65,450 ± 3111, P < 0.01). DS induced IR, similar to the traditionally used pharmacological factors, namely dexamethasone, TNF-α, and palmitate, which can be attributed to the significantly higher pro-inflammatory cytokines levels (TNF-α (2.28 ± 0.03 pg/mL vs. 2.38 ± 0.03 pg/mL, P < 0.01), interleukin 6 (IL)-6 (1.94 ± 0.02 pg/mL vs. 2.17 ± 0.04 pg/mL, P < 0.01), IL-17 (2.16 ± 0.02 pg/mL vs. 2.22 ± 0.002 pg/mL, P < 0.05), and interferon gamma (IFN-γ) (2.07 ± 0.02 pg/mL vs. 2.15 ± 0.04 pg/mL, P < 0.05)) in DS.Conclusions:DS can be explored as a novel inducer of IR in in vitro studies with further standardization, substituting the conventionally used pharmacological factors. Our findings also affirm the validity of ADMSC-CM as a prospective insulin sensitizer for T2DM therapy.
简介:摘要目的探讨CDC25B和14-3-3δ蛋白在膀胱癌中的表达与病理分级、临床分期的关系,为临床上判断膀胱癌的恶性程度和膀胱癌的新的治疗方法提供理论依据。方法采用免疫印迹(WesternBlot)法检测64例膀胱癌、9例癌旁正常膀胱粘膜中CDC25B和14-3-3δ蛋白的表达。1)观察CDC25B和14-3-3δ蛋白在膀胱癌和癌旁正常膀胱粘膜中的表达情况;2)分析CDC25B和14-3-3δ蛋白在膀胱癌中的表达与病理分级和临床分期的关系。结果所测组织中均有CDC25B的表达,膀胱癌中呈高表达;并且在膀胱癌各病理分级、临床分期间的表达亦有差异。CDC25B随着肿瘤的分化程度降低而表达逐渐增加,CDC25B在浸润性膀胱癌中表达高于非浸润性膀胱癌。14-3-3δ蛋白在癌旁正常膀胱粘膜中大量表达,在膀胱癌中均呈低表达,但未见其与肿瘤病理分级有明显相关性。结论CDC25B在膀胱癌组织中表达明显高于癌旁正常膀胱粘膜,且CDC25B的高表达与膀胱癌的分化程度和浸润性密切相关;14-3-3δ蛋白在癌旁膀胱粘膜大量表达,而在膀胱癌组织中均呈低表达,但与肿瘤分化程度无相关性。CDC25B及14-3-3δ蛋白可能成为膀胱癌潜在的诊断标记和治疗靶点。
简介:摘要报道1例基因诊断明确的线粒体3-羟基-3-甲基戊二酰辅酶A合成酶缺乏症(HMGCSD),并对国内外已报道病例进行文献复习。先证者,女,7个月16 d,因"发热4 d,喘息3 h,呼吸困难、呻吟2 h"急诊入院,主要表现为脑病、肝大、肝损害、低酮性低血糖、高脂血症,于住院第3天死于呼吸循环衰竭。全外显子组测序示HMGCS2复合杂合变异:c.1061+1G>C与c.476G>T,结合患儿临床特点,可基因诊断HMGCSD。文献检索共收集HMGCSD相关文献13篇,共26例患儿,发病年龄3个月~6岁,发病诱因主要为能量摄入不足,主要表现为低酮性低血糖、肝大、肝损害等,尿4-羟基-6-甲基-2-吡喃酮增高可能有强烈提示意义,3例死亡。26例患儿共报道HMGCS2突变32种,主要突变类型为错义突变。本研究为国内确诊第2例HMGCSD,发现2个HMGCS2新变异,扩展了HMGCS2临床表型及突变谱。
简介:H3,ahomogeneousacidicpolysaccharidewasobtainedfromtheseedsofCuscutachinensisLam.Itsstructurewascharacterizedforthefirsttimebychemicalandspectroscopicmethodstobeahighlybranchedheteropolysaccharidewithmeanmolecularweightofmorethanlxl0^6.Itwascomposedof1,6-1inked-β-DGalp,1,4-linked-β-DGalp,1,4-1inked-β-DGalA,1,3,6-1inked-β-DGalpand1,2,4-1inkedRhap,withbranchingpointsatO-2or0-4of1,2,4-1inkedRhapand0-3of1,3,6-1inked-β-DGalp.Itssidechainsincluded1-1inkedAraf,1,5-1inkedArafand1,3,5-1inkedArafatO-3of1,6-1inkedGalpinthemainchain.
简介:AbstractLong-term and stable preservation of bacteriophages is of crucial importance. Although many efforts have been made in the past decades to explore the influence of external factors on bacteriophage preservation, there is still little understanding, and a systematic description is lacking. In this study, we explored the influence of different factors on the preservation of lytic bacteriophage VP3, one of the typing bacteriophages of Vibrio cholerae O1 biotype El Tor, and attempted to optimize its preservation. We examined external factors, including temperature, solution, and cryoprotectant, in stable cooling/freezing conditions or alternate cooling/freezing and thawing. We found that whether in Luria-Bertani (LB) medium or SM buffer, in terms of 20-week stable cooling or freezing, -20 °C was the most damaging while 4 °C, -80 °C, and -196 °C were protective. Thirteen cycles of alternate cooling/freezing and thawing caused a loss in the survival rates of bacteriophages. The addition of cryoprotectant, glycerol (30%, w/v) or dimethyl sulfoxide (DMSO, 10%, w/v) significantly improved the survival rates of bacteriophages preserved at -20 °C. However, at 4 °C, -80 °C, and -196 °C, the cryoprotectant effect was only slightly positive or even harmful. In summary, for bacteriophage VP3, the best preservation method is to directly preserve the bacteriophage stocks in LB medium at -80 °C or -196 °C instead of storing them in SM buffer or adding cryoprotectant. Our results provided insights into the external influencing factors on bacteriophage VP3 during preservation at low temperature and can be applied to the optimization of bacteriophage preservation in the future.