Plasma Levels of miR-125b-5p and miR-206 in Acute Ischemic Stroke Patients After Recanalization Treatment: A Prospective Observational Study

Introduction: Multiple microRNAs (miRNAs) participate in the response to hypoxic/ischemic and ischemia-reperfusion events. However, the expression of these miRNAs in circulation from patients with acute ischemic stroke (AIS) receiving recanalization treatment has not been examined, and whether they are associated with the severity and outcome of stroke is still unknown. Materials and methods: In this prospective cohort study, plasma levels of miR-125b-5p, miR-15a-3p, miR-15a-5p, and miR-206 were measured at 24 hours after thrombolysis with or without endovascular treatment in 94 patients with AIS, as determined by qRT-PCR. Stroke severity was assessed based on National Institutes of Health Stroke Scale (NIHSS) score and infarct lesion. Intracranial haemorrhage (ICH) was recorded. An unfavorable outcome was defined as a modified Rankin Scale score greater than 2 at day 90 after stroke. Results: miR-125b-5p and miR-206 levels were correlated with NIHSS scores (P = .014 and P = .002) and cerebral infarction volumes (P = .025 and P = .030). miR-125b-5p levels were significantly higher in patients with an unfavorable outcome than in patients with a favorable outcome (P = .002) and showed good diagnostic accuracy in discriminating the presence of an unfavorable outcome (area under the curve .735, 95% confidence interval .623-.829, P < .001). No association was found between different miRNAs and ICH. Conclusions: In AIS patients after thrombolysis with or without endovascular treatment, miR-125b-5p is a novel prognostic biomarker highly associated with an unfavorable outcome. miR-125b-5p and miR-206 levels are associated with stroke severity.     

Inhibiting hydrogen sulfide production in umbilical stem cells reduces their protective effects during experimental necrotizing enterocolitis

IntroductionUmbilical mesenchymal stem cells (USC) have been shown to reduce illness in animal models of necrotizing enterocolitis (NEC), possibly through the paracrine release of hydrogen sulfide (H₂S). We hypothesized that animals treated with USCs with inhibited H₂S synthesis would exhibit more severe disease.MethodsNEC was induced in five-day-old mouse pups by formula feeding and hypoxic and hypothermic stress. Experimental groups received intraperitoneal injection of either saline vehicle or 80,000cells/gram of one of the following cell types: USC, USCs with negative-control siRNA, or USCs with targeted siRNA inhibition of the H₂S-producing enzymes. Pups were monitored by clinical assessment and after euthanasia, intestine and lung histologic injury were scored. Tissue was homogenized, and concentrations of IL-6, IL-10, and VEGF were determined by ELISA. For statistical analysis, p < 0.05 was considered significant.ResultsAnimals treated with negative-control siRNA USCs were significantly improved compared to vehicle. Clinical sickness scores as well as intestinal and lung histologic injury scores in the targeted siRNA groups were significantly worse when compared to the negative-control siRNA group. IL-6, IL-10, and VEGF had varying patterns of expression in the different groups.ConclusionInhibition of H₂S production in USCs reduces the beneficial effects of these cells during therapy in experimental NEC.Level of evidenceAnimal studies are typically described as “foundational evidence” without a true level assigned.Type of studyAnimal Study.     

The Presence of Sinus Tract Adversely Affects the Outcome of Treatment of Periprosthetic Joint Infections

BackgroundA sinus tract may be encountered in patients with periprosthetic joint infection (PJI) and constitutes a major criterion for diagnosis. The aim of this study is to identify associated factors for the presence of sinus tract and outcome of 2-stage exchange arthroplasty in these patients.MethodsWe retrospectively reviewed all patients with PJI following hip and knee arthroplasty from 2000 to 2017. Of them, 161 patients with a sinus tract had a minimum follow-up of 1 year following 2-stage exchange arthroplasty. These patients were matched 1:2 with those without sinus tract by using propensity score matching. Treatment success was assessed using the modified Delphi criteria. A multiple logistic regression analysis was performed to determine the effect of sinus tract on the outcome and associated factors for the presence of sinus tract.ResultsFactors significantly associated with sinus tract included smoking (odds ratio [OR] = 1.83), hypothyroidism (OR = 1.62), hypoalbuminemia (OR = 1.52), hip joint involvement (OR = 1.43), and prior revision surgery (OR = 1.37). Patients with sinus tract had a significantly higher rate of failure compared to those without sinus tract (OR = 2.94).ConclusionThis study demonstrates that the presence of sinus tract in patients with PJI adversely affects the outcome of treatment of these patients. The presence of sinus tract may be a proxy for other issues such as poor periarticular soft tissue, the poor nutritional status of the host, and multiple prior operations. These findings need to be borne in mind when treating patients with PJI and a concomitant sinus tract.     

Outcomes of Superficial and Deep Irrigation and Debridement in Total Hip and Knee Arthroplasty

BackgroundFollowing total knee arthroplasty and total hip arthroplasty, wound-related problems and deep periprosthetic joint infection may present in a similar fashion. Irrigation and debridement (I&D) has a great role in management of patients with early infection. The question that often arises is how to tell the difference between superficial and deep infection. This study evaluated the role and outcomes of both superficial and deep I&D in patients with wound-related issues and/or suspected periprosthetic joint infection.MethodsA retrospective study was conducted evaluating patients who underwent I&D within 28 days of total joint arthroplasty. A total of 176 cases with a minimum of 1-year follow-up were identified, and clinical records were reviewed in detail. Reoperations included superficial (fascia not opened) or deep (fascia opened) I&D. Failure was defined as the need for further surgical intervention within 1 year of initial I&D.ResultsThe overall success for superficial I&D was 84.28% vs 68.86% for deep I&D. The success of both deep and superficial I&D was higher if the I&D was performed closer to the index surgery. Superficial I&D in patients with a positive joint aspiration or evidence of intraoperative purulence or those in whom had no subcutaneous fluid had higher failure rates.ConclusionSuperficial I&D is a viable option in patients with wound-related issues as long as joint aspiration is performed to rule out infection involving the prosthesis. If there are findings of no fluid or purulence, fascia may need to be opened and the deeper tissues explored.     

miR-125b-5p对喉鳞状细胞癌细胞能量代谢及增殖的影响

目的  探讨miR-125b-5p对喉鳞状细胞癌（laryngeal squamous cell carcinoma,LSCC）细胞能量代谢和增殖的影响及其可能的作用机制。方法 实验分为miR-125b-5p转染组（miR-125b-5p模拟物转染LSCC）和对照组（空质粒转染LSCC细胞）。采用RT-qPCR 检测miR-125b-5p在正常支气管上皮细胞和LSCC细胞中的表达,CCK-8和流式细胞术检测LSCC细胞增殖、凋亡情况,Western blot检测miR-125b-5p过表达后LSCC中HK2的表达,3H-2DG法和乳酸盐比色测定实验分别检测LSCC细胞葡萄糖消耗和乳酸产生的情况。结果 RT-qPCR实验结果显示,与正常支气管上皮细胞相比,LSCC细胞中miR-125b-5p的表达降低（0.68±0.03 vs 0.22±0.05,t=7.025,P=0.001）。CCK-8实验结果显示,转染72 h和96 h后,miR-125b-5p转染组LSCC细胞的增殖能力均较对照组明显降低（P<0.05）。流式细胞术检测结果显示,miR-125b-5p转染组LSCC细胞凋亡率较对照组升高 ［（37.52±2.34）% vs （12.46±3.52）%,t=7.025,P<0.001）］,但细胞集落形成能力降低（0.29±0.02 vs 1.02±0.03,t=5.689,P=0.005）;荧光素酶报告基因测定实验结果显示,miR-125b-5p转染组的荧光素酶活性低于对照组（0.32±0.03 vs 1.01±0.02,t=7.543,P=0.001）;Western blot法实验结果显示,miR-125b-5p转染组HK2蛋白表达水平较对照组降低（0.12±0.02 vs 0.75±0.03,t=5.875,P=0.023）;miR-125b-5p转染组葡萄糖消耗量［（3.85±0.86） dpm/mg vs （10.52±1.34） dpm/mg,t=6.118,P=0.005］以及乳酸产生量［（4.23±1.36） dpm/mg vs （10.96±2.45） dpm/mg,t=5.907,P=0.002］亦较对照组降低。结论 miR-125b-5p可能通过下调HK2表达降低喉鳞状细胞癌细胞的能量代谢和增殖能力,诱导细胞凋亡。     

MUC16 suppresses human and murine innate immune responses

ObjectiveMUC16, the mucin that contains the CA125 epitopes, suppresses the cytolytic responses of human NK cells and inhibits the efficacy of therapeutic antibodies. Here, we provide further evidence of the regulatory role of MUC16 on human and murine NK cells and macrophages.MethodsTarget cell cytolysis and doublet formation assays were performed to assess effects of MUC16 on human NK cells. The effect of MUC16 on ovarian tumor growth was determined in a mouse model by monitoring survival and ascites formation. Innate immune cells from spleens and peritoneal cavities of mice were isolated and stimulated in vitro with anti-CD40 antibody, lipopolysaccharide and IFN-γ and their ability to cytolyse MUC16 expressing and non-expressing cells was determined.ResultsWe confirm that MUC16 inhibits cytolysis by human NK cells as well as the formation of NK-tumor conjugates. Mice implanted with MUC16-knockdown OVCAR-3 show >2-fold increase in survival compared to controls. Murine NK cells and macrophages are more efficient at lysing MUC16-knockdown cells. In vitro cytotoxicity assays with NK cells and macrophages isolated from mice stimulated with anti-CD40 antibody showed 2–3-fold increased activity against the MUC16-knockdown cells as compared to matching target cells expressing this mucin. Finally, knockdown of MUC16 increased the susceptibility of cancer cells to ADCC by murine splenocytes.ConclusionsFor the first time, we demonstrate the immunoregulatory effects of MUC16 on murine NK cells and macrophages. Our study implies that the immunoregulatory role of MUC16 on murine NK cells and macrophages should be considered when examining the biology of MUC16 in mouse models.     

肝硬化患者血清CA125水平升高的相关因素分析

背景　肝硬化患者血清CA125呈现不同程度的升高,但肝硬化血清CA125升高的影响因素尚不明确。从多角度分析影响肝硬化患者血清CA125升高的相关因素非常必要。目的　探讨影响肝硬化患者血清CA125水平升高的相关因素。方法　选取2012年1月-2018年2月于郑州大学第二附属医院住院的肝硬化患者205例,行腹腔穿刺113例。分组情况：根据血清CA125水平分为CA125正常组（n=39）与CA125异常组（n=166）;根据腹腔积液量分为无腹腔积液组（n=52）、少量腹腔积液组（n=40）、中量腹腔积液组（n=51）、大量腹腔积液组（n=62）;根据Child-Pugh分级分为Child-Pugh A级组（5~6分）46例、Child-Pugh B级组（7~9分）87例、Child-Pugh C级组（≥10分）72例;根据腹腔积液有核细胞计数分为腹腔积液有核细胞计数正常组65例、腹腔积液有核细胞计数升高组48例（共113例行腹腔穿刺）。比较CA125异常组与CA125正常组的总胆红素、总胆汁酸、丙氨酸氨基转移酶（ALT）、天冬氨酸氨基转移酶（AST）、凝血酶原时间（PT）、清蛋白、总蛋白、凝血酶原活动度、年龄、性别;观察腹腔积液量与血清CA125水平的相关性、Child-Pugh分级与血清CA125水平的相关性、腹腔积液有核细胞计数与血清CA125水平的相关性。结果　CA125异常组总胆红素、总胆汁酸、AST高于CA125正常组,PT长于CA125正常组,清蛋白、总蛋白、凝血酶原活动度低于CA125正常组（P<0.05）。无腹腔积液组血清CA125水平低于少量腹腔积液组、中量腹腔积液组、大量腹腔积液组（P<0.05）;少量腹腔积液组血清CA125水平低于中量腹腔积液组、大量腹腔积液组（P<0.05）。Child-Pugh A级组血清CA125水平低于Child-Pugh B级组、Child-Pugh C级组（P<0.05）;Child-Pugh B级组血清CA125水平低于Child-Pugh C级组（P<0.05）。腹腔积液有核细胞计数升高组血清CA125水平高于腹腔积液有核细胞计数正常组（P=0.023）。腹腔积液量、Child-Pugh分级、腹腔积液有核细胞计数与血清CA125水平均相关（P<0.05）。结论　血清CA125水平升高与腹腔积液量、Child-Pugh分级及腹腔积液有核细胞计数相关,其可用来评估腹腔积液量的多少、肝功能受损程度及出现腹膜炎并发症的可能性。     

四项血清肿瘤标志物联合检测和ROMA指数在卵巢癌中的应用价值

目的 探讨四项血清肿瘤标志物HE4、CEA、CA125、CA72-4的联合检测和ROMA指数在卵巢癌中的应用。方法 选择55例卵巢癌患者作为恶性组,61例卵巢良性肿瘤患者作为良性组,60例体检合格的健康人群作为对照组,检测三组的HE4、CEA、CA125和CA72-4水平,并利用HE4和CA125的值计算ROMA指数,比较三组上述参数的水平、阳性率、灵敏度及特异度。绘制ROC曲线以评价上述四项血清肿瘤标志物单项和联合检测及ROMA指数对卵巢癌的诊断价值。结果 恶性组中的HE4、CEA、CA125、CA72-4的单项及联合检测和ROMA指数水平及阳性率均高于良性组和对照组,差异有统计学意义(P＜0.05)。HE4、CEA、CA125和CA72-4四项肿瘤标志物联合检测后,灵敏度由单项CA125最高54.5%提高到78.2%,再联合ROMA指数,灵敏度进一步提高,为92.7%。对ROC曲线分析,HE4、CEA、CA125、CA72-4和四项联合检测及ROMA指数的曲线下面积分别为0.820、0.706、0.774、0.736、0.839、0.880,可知ROMA指数对卵巢癌诊断的准确性最好。结论 四项血清肿瘤标志物的联合检测以及ROMA指数对卵巢癌的诊断具有重要意义。