Specific primer amplification of the VP1 region directed by 5′ UTR sequence analysis: Enterovirus testing and identification in clinical samples from hand-foot-and-mouth disease patients

Many genotypes of the enterovirus (EV) pathogens can cause clinical hand-foot-and-mouth disease (HFMD). Therefore, rapid identification and monitoring of HFMD pathogens can be difficult, especially from the original clinical specimens. In this study, both universal pan-enterovirus and EV71/CA16 VP1-specific primer sets were designed and used to examine clinical specimens from HFMD patients. Based on the initial sequence analysis of the 5′-untanslated region (5′-UTR) and VP1 amplification products, additional primers for the VP1 region were redesigned for further genotyping of the remaining small portion non-EV71/non-CA16 specimens. With a known panel, it was possible to identify 15 out of 16 members using 5′-UTR sequence typing and VP1 typing, suggesting good detectability and genotyping of this method. One strain that was not typed by 5′-UTR was shown to be a recombinant virus. When this method was applied to examine clinical specimens from 44 suspected HFMD patients, 41 were detected as EV positive. In only one case, the VP1 sequence could not be identified. Four types of EVs, including CA16 (26/41, 63.4%), EV71-C4 (6/41, 14.6%), CA6 (5/41, 12.2%) and CA10 (3/41, 7.3%), were detected. In conclusion, 5′ UTR amplification sequencing and subsequent VP1 specific primer amplification ensures a high detection rate and good genotyping accuracy in the examination of clinical samples. This detection strategy can be used for routine evaluation and monitoring of HFMD to follow local trends of EV infection.     

SRC-3 coactivator regulates cell resistance to cytotoxic stress via TRAF4-mediated p53 destabilization

Steroid receptor coactivator 3 (SRC-3) is an oncogenic nuclear receptor coactivator that plays a significant role in drug resistance. Using a lentiviral cDNA library rescue screening approach, we identified a SRC-3 downstream gene—TRAF4 (tumor necrosis factor [TNF] receptor associated-factor 4)—that functions in cell resistance to cytotoxic stress. TRAF4 expression is positively correlated with SRC-3 expression in human breast cancers. Similar to that observed for SRC-3 overexpression, breast cancer cells overexpressing TRAF4 are more resistant to stress-induced death. Here, we further dissected the underlying molecular mechanism for SRC-3 and TRAF4-mediated resistance to cytotoxic agents. We observed that SRC-3 expression is inversely correlated with the expression of p53-regulated proapoptotic genes in breast cancers and further found that SRC-3 and TRAF4 overexpression diminished cytotoxic stress-induced up-regulation of the tumor suppressor p53 protein. To determine the mechanism, we showed that the TRAF domain of TRAF4 bound to the N-terminal TRAF-like region of the deubiquitinase HAUSP (herpesvirus-associated ubiquitin-specific protease; also named USP7) and blocked the access of p53 to the same region of HAUSP. This TRAF4-mediated inhibition of HAUSP then led to the loss of p53 deubiquitination and its stabilization in response to cellular stress. Consistent with this cellular function, we also found that TRAF4 overexpression in breast cancer patients was associated significantly with poor prognosis. Because of SRC-3''s ability to abrogate p53 function, our results suggest that SRC-3 overexpression may be especially important in tumors in which p53 is not mutated.     

间变脑胶质瘤中异柠檬酸脱氢酶突变对患者术后替莫唑胺化学 治疗及预后的影响：285例单中心回顾性研究

目的 分析间变脑胶质瘤中异柠檬酸脱氢酶（IDH）突变与患者临床病理学因素的相关性 及对术后化学治疗敏感性的影响。方法 回顾性连续纳入中国胶质瘤基因图谱计划（CGGA）数据库中 接受开颅手术且术后病理诊断为间变脑胶质瘤（世界卫生组织Ⅲ级）患者,术后接受放射治疗和（或）替 莫唑胺（TMZ）化学治疗。肿瘤组织标本在放射治疗和（或）化学治疗前通过手术切除获得,使用QIAamp DNA Mini 试剂盒按照说明书从冷冻肿瘤组织中提取基因组DNA,使用纳米滴ND-1000 分光光度计测 定DNA 浓度和质量,进行IDH1/2 突变、1p/19q 缺失、MGMT 启动子甲基化的检测。应用SPSS 16.0 软件 通过单因素和多因素Cox 生存分析分析临床及分子病理因素与患者总生存期的相关性及对术后放射 治疗和替莫唑胺化学治疗敏感性的影响;使用GraphpadPrism 8 软件进行Kaplan-Meier 曲线生存分析。 结果 共纳入285 例间变脑胶质瘤患者,其中201 例（70.5%）表现为IDH 突变,84 例（29.5%）表现为IDH 野生。多因素Cox 回归分析显示,IDH突变（HR=0.531,95%CI：0.385～0.733,P ＜ 0.001）及1p/19q 联合 缺失（HR=0.352,95%CI：0.179～0.691,P=0.002）均可独立预测患者总生存期,而O6-甲基鸟嘌呤-DNA 甲基转移酶（MGMT）启动子甲基化的预测价值差异无统计学意义（P=0.071）。IDH 突变组中,接受术后 TMZ化学治疗的患者较未接受者预后有所改善,但差异无统计学意义（中位生存期：1 793 d 比1 455 d, P=0.059）,而在IDH野生组中,TMZ化学治疗对患者预后改善不明显（中位生存期：535 d 比 415 d, P=0.890）。结论 本研究发现IDH突变型间变脑胶质瘤在接受术后TMZ化疗后,预后有改善趋势,相关 结果可能为今后WHO Ⅲ级脑胶质瘤术后化疗的研究提供思路。     

Combination of Radiological and Gray Level Co-occurrence Matrix Textural Features Used to Distinguish Solitary Pulmonary Nodules by Computed Tomography

The objective of this study was to investigate the method of the combination of radiological and textural features for the differentiation of malignant from benign solitary pulmonary nodules by computed tomography. Features including 13 gray level co-occurrence matrix textural features and 12 radiological features were extracted from 2,117 CT slices, which came from 202 (116 malignant and 86 benign) patients. Lasso-type regularization to a nonlinear regression model was applied to select predictive features and a BP artificial neural network was used to build the diagnostic model. Eight radiological and two textural features were obtained after the Lasso-type regularization procedure. Twelve radiological features alone could reach an area under the ROC curve (AUC) of 0.84 in differentiating between malignant and benign lesions. The 10 selected characters improved the AUC to 0.91. The evaluation results showed that the method of selecting radiological and textural features appears to yield more effective in the distinction of malignant from benign solitary pulmonary nodules by computed tomography.     

Successful recovery of COVID-19-associated recurrent diarrhea and gastrointestinal hemorrhage using convalescent plasma

Background:Gastrointestinal symptoms are not rare among coronavirus disease 2019(COVID-19)patients,but there have been no reports regarding convalescent plasma therapy for the recovery of gastrointestinal problems in COVID-19 patients.Case presentation:We present two cases of patients with COVID-19-associated recurrent diarrhea and positive fecal occult blood who successfully recovered after a one-time convalescent plasma administration.Conclusion:When COVID-19 patients develop recurrent or refractory gastrointestinal symptoms and fail to respond to the available treatment,alternative therapy with convalescent plasma administration may be considered.     

Association of a Tgf‐B1‐509c/T Polymorphism with Gastric Cancer Risk: A Meta‐Analysis

Published data on the association between the transforming growth factor B1 (TGF‐B1) gene 509C/T polymorphism and gastric cancer risk are inconclusive. To derive a more precise estimation of the relationship, a meta‐analysis of the TGFB1‐509C/T polymorphism (with 2130 cases and 2374 controls) from seven published case‐control studies was performed. Crude odds ratios (ORs) with 95% confidence intervals (CIs) were used to assess the strength of association in the codominant model, the dominant model, and the recessive model. In the overall analysis, the T allele was significantly associated with susceptibility to gastric cancer in the recessive model (TT vs. CC+CT) (TT vs. CC+CT: OR = 1.35, 95% CI: 1.10–1.66, P = 0.10 for heterogeneity) when all the included studies were pooled into the meta‐analysis. In the stratified analysis by country, the T allele was also found to be significantly associated with increased gastric cancer risk in the recessive model (TT vs. CC+CT) in Chinese studies and in T versus C in the Indian study. In conclusion, this meta‐analysis supports the TGFB1‐509T polymorphism as a susceptibility factor for gastric cancer.     

心脏MR特征追踪技术早期评价高血压性心脏病左心室亚临床功能障碍的研究

目的探讨心脏MR特征追踪(CMR-FT)技术在早期定量分析高血压性心脏病(HHD)左心室亚临床功能障碍中的应用价值。方法回顾性分析2018年10月至2019年11月于华中科技大学同济医学院附属同济医院诊治的高血压性心脏病伴左心室肥厚(HHD-LVH)患者16例、高血压性心脏病不伴左心室肥厚(HHD-非LVH)患者24例及年龄与性别相匹配的血压正常者24名。所有研究对象进行3.0 T CMR检查,扫描序列包括心脏电影和钆延迟增强。通过Argus后处理软件测量左心室心功能参数,主要包括左心室舒张末期容积指数(LVEDVI)、左心室质量指数(LVMI)、左心室壁最大厚度(LVMWT)及左心室质量与左心室舒张末期容积比例(M/V)。采用CVI42商业后处理软件测量左心室心肌应变指标,包括整体径向、圆周、长轴应变(GRS、GCS、GLS),收缩期径向、圆周、长轴应变率峰值(SRSR_peak、SCSR_peak、SLSR_peak),舒张期径向、圆周、长轴应变率峰值(DRSR_peak、DCSR_peak、DLSR_peak)。以单因素方差分析(scheffe校正)或Kruskal-Wallis检验比较3组组间连续性变量的差异。以Pearson或Spearman相关系数进行相关性分析。结果HHD-LVH组的LVEDVI、LVMI、LVMWT及M/V均显著高于HHD-非LVH组和对照组(P<0.05)。与对照组比较,HHD-LVH组的GRS、GCS和GLS减低,HHD-LVH组和HHD-非LVH组的DRSR_peak、DCSR_peak和DLSR_peak均减低,差异均具有统计学意义(P<0.05)。相关性分析发现LVMI与GRS(r=-0.384,P=0.002)、GCS(r=0.392,P=0.001)、GLS(r=0.491,P<0.0001)存在线性相关,LVMWT与GRS(r=-0.362,P=0.003)、GCS(r=0.384,P=0.002)、GLS(r=0.422,P=0.001)存在非线性相关,LVEDVI与GRS(r=-0.295,P=0.018)、GCS(r=0.264,P=0.035)存在非线性相关。结论CMR-FT技术测量的心肌应变指标可作为早期评价HHD患者左心室亚临床功能障碍的影像学参数,并有助于指导早期干预治疗,改善心室重塑。     

肌间隙入路椎弓根置钉治疗胸腰段脊柱骨折的临床效果

目的 探讨肌间隙入路(Wiltse入路)椎弓根置钉治疗胸腰段脊柱骨折的临床效果.方法 前瞻性研究2017年1月—2019年10月太仓市第一人民医院骨科收治的100例胸腰段脊柱骨折患者,按随机数字表法分为Wiltse入路组和常规入路组各50例.Wiltse入路组男性29例,女性21例;年龄18～60岁,平均42.2岁;高处坠落伤19例,道路交通伤25例,砸伤6例.常规入路组男性32例,女性18例;年龄18～60岁,平均41.5岁;高处坠落伤17例,道路交通伤28例,砸伤5例.比较两组患者相关康复指标、手术效果、恢复情况及术后切口浅表感染、腰背疼痛等并发症发生情况.结果 Wiltse入路组术中出血量、手术时间、住院时间均少于常规入路组[(119.8±8.3)mL vs.(192.2±10.5)mL、(90.7±6.4)min vs.(119.1±8.7)min、(11.8±1.6)d vs.(15.1±2.0)d,P<0.05].Wiltse入路组术前、术后12个月椎体前缘高度分别为(15.5±1.8)mm、(26.1±3.0)mm,常规入路组分别为(15.2±1.1)mm、(25.3±3.1)mm,两组术后12个月椎体前缘高度较术前均提高(P<0.05);Wiltse入路组术前、术后12个月矢状位椎体Cobb角分别为(23.2±2.5)°、(5.9±0.7)°,常规入路组分别为(22.7±3.3)°、(6.3±1.0)°,两组术后12个月矢状位椎体Cobb角度较术前均降低(P<0.05);两组术后12个月椎体前缘高度、矢状位椎体Cobb角度比较差异均无统计学意义(P>0.05).Wiltse入路组术前、术后6个月Oswestry功能障碍指数问卷表(ODI)评分分别为(40.7±5.2)分、(5.0±0.8)分,常规入路组分别为(39.8±6.4)分、(5.2±0.7)分,两组术后6个月ODI评分较术前均降低(P<0.05),且术后6个月ODI评分两组间比较差异无统计学意义(P>0.05).Wiltse入路组术后总并发症发生率低于常规入路组(2％vs.16％,P<0.05).结论 胸腰段脊柱骨折中实施Wiltse入路椎弓根置钉治疗可减少术中出血量,缩短手术时间及住院时间,其解剖结构及脊柱功能恢复效果与常规入路治疗相当,但可减少术后并发症的发生.     

术前3D数字化技术模拟对胫骨平台骨折手术操作、术后关节功能恢复的影响

目的 探讨术前3D数字化技术模拟手术复位及操作对复杂胫骨平台骨折患者的手术效果及关节功能恢复的影响.方法 前瞻性分析2016年2月—2018年8月新疆医科大学第六附属医院骨病矫形外科收治的92例复杂型胫骨平台骨折患者资料,随机数字表法分为3D模拟组(术前CT扫描数据导入Minics软件内进行三维重建模拟骨折复位、手术入路等操作)、对照组(常规手术治疗)各46例.3D模拟组男性28例,女性18例;年龄26～62岁,平均42.2岁.对照组男性26例,女性20例;年龄26～65岁,平均41.1岁.患者分别于术后4、6个月到门诊接受膝关节功能测定,比较两组患者的手术操作相关指标、术后膝关节功能、胫骨平台内翻角(TPA)、胫骨平台后倾角(PA)的变化及手术并发症.结果 3D模拟组患者的手术时间、术中出血量、术中透视时间均少于对照组[(108.6±16.2)min vs.(119.4±20.0)min、(180.5±36.1)mL vs.(224.3±40.6)mL、(90.6±11.7)min vs.(102.5±14.3)min,P<0.05],3D模拟组患者的解剖复位率高于对照组(93.48％vs.78.26％,P<0.05);术后4个月,3D模拟组患者的Lysholm评分高于对照组[(78.33±8.61)分vs.(74.18±9.25)分,P<0.05],术后6个月,3D模拟组患者的Lysholm评分与对照组比较差异无统计学意义(P>0.05);手术后即刻、术后4、6个月,3D模拟组和对照组的膝关节TPA、PA角比较差异无统计学意义(P>0.05);3D模拟组的手术并发症发生率4％与对照组9％比较,差异无统计学意义(P>0.05).结论 术前3D数字化技术模拟手术复位及操作有利于减少复杂胫骨平台骨折患者手术操作的难度,提高解剖复位率,促进早期膝关节功能的恢复.