Association of interleukin-6 methylation in leukocyte DNA with serum level and the risk of ischemic heart disease

Background Interleukin-6 (IL-6), a multifunctional cytokine, plays an important role in the development of ischemic heart disease (IHD), and DNA hypomethylation of 2 CpGs, located downstream in the proximity of the IL-6 gene promoter, has been associated with risk factor for IHD. This study was to examine the association of blood leukocyte DNA methylation of the 2 CpGs in IL-6 with the risk of IHD and the serum IL-6 level. Methods IL-6 methylation levels of 582 cases and 673 controls were measured using the bisulfite pyrosequencing technology. Serum level of IL-6 was measured using enzyme-linked immunosorbent assay. Results The IL-6 methylation was significantly lower in IHD cases than in the controls, irrespective of CpG site. After multivariate adjustment, lower (< median) average IL-6 methylation was associated with an increased risk of IHD (OR 1.57, 95% CI 1.22–2.02, p?β?=??1.02?pg/mL per increase in IL-6 methylation, p?=?0.002) among IHD cases. This significant relationship was not observed among controls. Conclusions DNA hypomethylation of IL-6 gene measured in blood leukocytes was associated with increased risk of IHD. IL-6 demethylation may upregulate its expression, whereby exerting its risk effect on the development of IHD.     

脑梗死患者血清和脑脊液S-100B蛋白含量的变化

目的：通过测定脑梗死患者急性期，恢复期血清和脑脊液S-100B蛋白含量动态变化，探讨其在脑梗死发病中的临床意义。方法：60例脑梗死住院患者（梗死组），将发病72h和7、21d的脑脊液及静脉血测定血清和脑脊液中S-100B蛋白含量，并分别与正常组比较。结果：梗死组患者发病72h和7d时血清和脑脊液S-100B蛋白显著高于21d患者和正常组（P〈0．01）；血清和脑脊液S-100B蛋白含量与梗死体积及神经功能缺损程度均呈正相关（P〈0．01）；脑脊液S-100B蛋白含量与年龄正相关，血清S-100B蛋白含量与年龄无关。结论：血清或脑脊液中的S-100B蛋白浓度可反映神经胶质细胞的损害程度。对判断病情程度。评估预后和调整治疗方案有重要意义。     

Amniotic epithelial cells promote wound healing in mice through high epithelialization and engraftment

Although human amniotic epithelial cells (AMEs) are an attractive source of stem cells, their therapeutic potential in wound healing has not been fully investigated. We evaluated the therapeutic potential of AMEs for wound healing. Real‐time PCR showed that the epithelialization growth factors epidermal growth factor (EGF), platelet‐derived growth factor (PDGF)‐B and chemotactic factors interleukin‐8 (IL‐8 or CXCL8) and neutrophil‐activating protein‐2 (NAP‐2 or CXCL7) were upregulated in AMEs compared with adipose‐derived mesenchymal stem cells (ADMs). In vitro scratch wound assays revealed that AME‐derived conditioned medium substantially accelerated wound closure. Wounds in NOD/SCID mice were created by skin excision, followed by AME transplantation. AMEs implantation significantly accelerated wound healing and increased cellularity and re‐epithelialization. Transplanted AMEs exhibited high engraftment rates and expressed keratinocyte‐specific proteins and cytokeratin in the wound area, suggesting direct benefits for cutaneous closure. Taken together, these data indicate that AMEs possess therapeutic capability for wound healing through the secretion of epithelialization growth factors and enhanced engraftment properties. Copyright © 2015 John Wiley & Sons, Ltd.     

Cell-specific roles of GRK2 in onset and severity of hypoxic-ischemic brain damage in neonatal mice

The ubiquitously expressed kinase GRK2 protects against cellular overstimulation by desensitizing G protein-coupled receptors and regulating intracellular signaling. Recently, we described that hypoxia-ischemia (HI)-induced brain damage was accelerated and increased in GRK2^+/? neonatal mice. Using Cre-Lox technology we now investigated the role of decreased GRK2 in only microglia/macrophages or forebrain neurons in development of HI brain injury.Low GRK2 in microglia/macrophages (LysM-GRK2^f/+ mice) was sufficient to accelerate onset of HI damage, without affecting the severity of brain injury at 24 h post-HI as compared to LysM-GRK2^+/+ littermates. Consistently, the ipsilateral hemisphere of GRK2^+/? mice contained microglia with a more rounded phenotype compared to WT mice at 3 h post-HI. Inhibition of microglial/macrophage activity by minocycline treatment prevented the early onset of HI injury in GRK2^+/? mice. In vitro, primary GRK2^+/? microglia stimulated with LPS produced more TNF-α than WT microglia via a p38-dependent pathway. In vivo, HI-induced cerebral p38 activation and TNF-α production were increased in GRK2^+/? mice or in LysM-GRK2^f/+ mice. Our findings indicate that low GRK2 in microglia/macrophages accelerates brain damage via a GRK2/p38/TNF-α-dependent pathway.Reduced GRK2 only in forebrain neurons (CamKIIα-GRK2^f/+ mice) significantly increased severity of HI brain damage without affecting the onset of brain damage.In conclusion, our data indicate that low GRK2 in microglia/macrophages facilitates activation of these cells which may contribute to the earlier onset of cerebral HI injury associated with increased p38 phosphorylation and TNF-α production. The level of GRK2 in neurons is crucial for determining the ultimate severity of HI damage in the newborn brain.     

Cellular basis of diabetic nephropathy: V. Endoglin expression levels and diabetic nephropathy risk in patients with Type 1 diabetes

Endoglin is an accessory receptor molecule that, in association with transforming growth factor β (TGF-β) family receptors Types I and II, binds TGF-β1, TGF-β3, activin A, bone morphogenetic protein (BMP)-2 and BMP-7, regulating TGF-β dependent cellular responses. Relevant to diabetic nephropathy, endoglin, expressed in vascular endothelial and smooth muscle cells, fibroblasts, and mesangial cells, negatively regulates extracellular matrix (ECM). The aim of this study was to evaluate endoglin expression in cultured skin fibroblasts from patients with Type 1 diabetes with and without diabetic nephropathy. Kidney and skin biopsies were performed in 125 Type 1 diabetic patients. The 20 with the fastest rate of mesangial expansion (estimated by electron microscopy) and proteinuria (“fast-track”) and the 20 with the slowest rate and normoalbuminuria (“slow-track”), along with 20 controls were studied. Endoglin mRNA expression was assessed by microarray and quantitative real-time polymerase chain reaction (QRT-PCR) and protein expression by Western blot. Age and sex distribution were similar among groups. Diabetes duration was similar (20±8 vs. 24±7 years), hemoglobin A1c lower (8.4±1.2% vs. 9.4±1.5%), and glomerular filtration rate higher (115±13 vs. 72±20 ml/min per 1.73 m²) in slow-track vs. fast-track patients. Microarray endoglin mRNA expression levels were higher in slow-track (1516.0±349.9) than fast-track (1211.0±274.9; P=.008) patients or controls (1223.1±422.9; P=.018). This was confirmed by QRT-PCR. Endoglin protein expression levels correlated with microarray (r=0.59; P=.044) and QRT-PCR (r=0.61; P=.034) endoglin mRNA expression. These studies are compatible with the hypothesis that slow-track Type 1 diabetic patients, strongly protected from diabetic nephropathy, have distinct cellular behaviors that may be associated with reduced ECM production.     

Toluidine blue-mediated photodynamic therapy of oral wound infections in rats

The purpose of this study was to examine the effect of toluidine blue (TB)-mediated photodynamic therapy (PDT) on oral wound infections in rats. The study called for a combination treatment of a 1mg/ml solution of TB with a red light at three intensity settings of 12 J/cm², 24 J/cm² and 48 J/cm². In the group that was given the highest light dose of 48 J/cm², an average kill rate of approximately 97% was achieved. A lesser killing effect was achieved in the group that was subjected to the lowest light dose of 12 J/cm², where an average of approximately 25% of the bacteria survived. After PDT, the lesions were allowed to develop, and the peak size of the lesions was larger in the control group than in the test groups, especially for the 48 J/cm² group. We also observed that in the 24 J/cm² and 48 J/cm² groups the lesions were of significantly smaller size. Our study demonstrated that combined TB-PDT therapy can successfully treat oral wound infections in rats. These promising results recommend the use of this treatment as a possible alternative to topical anti-microbials in future clinical applications.     

PUMA、caspase-3蛋白在胃癌组织中的表达及其临床意义

目的通过检测PUMA、caspase-3蛋白在胃癌组织和癌旁正常组织中的表达水平,从而探讨两者在胃癌发生、发展中的作用及其临床意义。方法针对40例胃癌组织标本及30例癌旁正常组织标本,通过免疫组化方法对其PUMA、caspase-3蛋白的表达进行测定。结果 PUMA蛋白在胃癌组织中的表达阳性率为22.5%(9/40),在癌旁正常胃组织中表达阳性率为66.7%(20/30),两者比较差异具有统计学意义;caspase-3在胃癌组织中的表达阳性率为25.0%(10/40),在癌旁正常胃组织中表达阳性率为70.0%(21/30),两者比较差异具有统计学意义。PUMA的表达与临床病理分期、肿瘤病理分化程度、是否伴有淋巴结转移有关,PUMA与caspase-3蛋白在胃癌组织中表达呈正相关。结论 PUMA与caspase-3蛋白在胃癌的发生发展中起重要作用,可能会成为胃癌的分子标记物或胃癌治疗的新分子靶点。     

类风湿性关节炎灭活滑膜液细胞对非灭活滑膜液细胞γ-干扰素影响的初步研究

目的观察类风湿性关节炎(rheumatoid arthritis,RA)灭活滑膜液细胞对非灭活滑膜液细胞IFN-γ水平的影响。方法抽取RA患者膝关节液分离滑膜液细胞,分实验组:2.0×10~6/mL非灭活滑膜液细胞,分别与2.0×10~5/mL、4.0×10~4/mL、2.0×10~4/mL甲醛灭活后的滑膜液细胞(灭活滑膜液细胞比非灭活滑膜液细胞为1:10、1:50、1:100)混合培养;对照组:单独培养细胞数为2.0×10~6/mL的非灭活滑膜液细胞。37℃,5%CO_2条件下培养48 h后,ELISA法测定炎症因子IFN-γ水平。结果(1)实验组较对照组IFN-γ水平明显降低(P<0.05);(2)实验组各组间比较,灭活滑膜液细胞比非灭活滑膜液细胞为1:10组较1:50组及1:100组相比较IFN-γ水平显著降低(P<0.05)。1:50组同1:100组IFN-γ水平无显著性差异(P>0.05)。结论(1)灭活滑膜液细胞对非灭活滑膜液细胞IFN-γ的分泌具有抑制作用;(2)灭活滑膜液细胞比非灭活滑膜液细胞为1:10时,抑制IFN-γ分泌的作用最强;(3)灭活滑膜液细胞可抑制非灭活滑膜液的IFN-γ分泌,其机制有待进一步研究。     

Is gastrointestinal dysfunction induced by gastric cancer peritoneal metastasis relevant to impairment of interstitial cells of Cajal?

Although impaired gastrointestinal motility from gastric cancer peritoneal metastasis (GCPM) causes extraordinary pain, its cause is unclear. Interstitial cells of Cajal (ICC) are apparently pacemaker cells, and their loss could cause motor dysfunction. In this study, we developed a mouse model for GCPM, and investigated electrophysiological changes in the small intestine and attendant changes in ICC. We found decreased ICC and disrupted electrical rhythm in the model. Pathologic ICC changes were well described. Cancer peritoneal metastasis may impair intestinal myoelectrical activity by damaging ICC and ICC networks. Interstitial cells of Cajal will be a target of palliative treatment and merit further study.