凉血通瘀方干预高血压大鼠急性脑出血模型对脑组织中差异miRNA表达的影响＊

目的 研究凉血通瘀方对高血压大鼠急性脑出血模型脑组织miRNA表达的影响，对差异表达的miRNA靶基因进行分析，探索凉血通瘀方可能的药效机制。方法 将自发性高血压大鼠随机分成对照组（B）和实验组（C）。适应性饲养一周后，C组灌胃凉血通瘀方，B组灌胃等体积生理盐水，连续5天，每天1次。构建脑出血模型后收集脑组织，借助全转录组测序技术获得miRNA表达量，与miRBase数据库比对获取已知miRNA，使用miRDeep2预测新miRNA。差异分析软件为DESeq2，筛选阈值为|log₂FC| ≥1 并且P <0.05。对显著差异表达的miRNA进行靶基因预测，对靶基因进行GO功能、KEGG通路富集和PPI网络分析。结果 实验组和对照组对比，共发现21个显著差异表达的miRNA，上调有9个，下调有12个，共预测得到1243个有统计学意义的靶基因。GO富集分析发现，生物过程中突触囊泡分泌的调节、神经递质分泌的调节和神经递质运输的调节占前三位，神经元投射终点、全膜、质膜区域和细胞投射则是主要的细胞成分。分子功能分别为小GTPase绑定、底物特异性跨膜转运蛋白活性和离子跨膜转运体活性。通路分析结果显示，靶基因在癌证通路、pI3K-Akt信号通路、人类乳头瘤病毒感染、神经活性配体-受体相互作用和MAPK通路等分布广泛。采用STRING网站和Cytoscape软件，根据MCC算法筛选出ADRA2C、CASR、CCL28、CCR1、DRD2、GNAT3、GRM2、DYNC1LI1、GABBR1、GNAI1等核心靶基因。结论 凉血通瘀方对脑出血急性期鼠脑组织内miRNA的表达有重要影响；显著差异表达miRNAs可能通过靶向核心基因调控凉血通瘀方干预急性脑出血的病理过程及预后。     

Biomarkers of in vivo limbal stem cell function

PurposeTo evaluate in vivo parameters as biomarkers of limbal stem cell function and to establish an objective system that detects and stage limbal stem cell deficiency (LSCD).MethodsA total of 126 patients (172 eyes) with LSCD and 67 normal subjects (99 eyes) were included in this observational cross-sectional comparative study. Slit-lamp biomicroscopy, in vivo laser scanning confocal microscopy (IVCM), and anterior segment optical coherence tomography (AS-OCT) were performed to obtain the following: clinical score, cell morphology score, basal cell density (BCD), central corneal epithelial thickness (CET), limbal epithelial thickness (LET), total corneal nerve fiber length (CNFL), corneal nerve fiber density (CNFD), corneal nerve branch density (CNBD), and tortuosity coefficient. Their potential correlations with the severity of LSCD were investigated, and cutoff values were determined.ResultsAn increase clinical score correlated with a decrease in central cornea BCD, limbal BCD, CET, mean LET, maximum LET, CNFL, CNFD, CNBD, and tortuosity coefficient. Regression analyses showed that central cornea BCD, CET and CNFL were the best parameters to differentiate LSCD from normal eyes (Coef = 3.123, 3.379, and 2.223; all p < 0.05). The rank correlation analysis showed a similar outcome between the clinical scores and the central cornea BCD (ρ = 0.79), CET (ρ = 0.82), and CNFL (ρ = 0.71). A comprehensive LSCD grading formula based on a combination of these parameters was established.ConclusionsA comprehensive staging system combining clinical presentation, central cornea BCD, CET, and CNFL is established to accurately and objectively diagnose LSCD and stage its severity.     

Cribado de cáncer de pulmón: Supervivencia en un amplio programa de detección precoz en España (I-ELCAP)

IntroductionLung cancer (LC) is usually diagnosed at advanced stages with only a 12% 5-year survival. Trials as NLST and NELSON show a mortality decrease, which justifies implementation of lung cancer screening in risk population. Our objective was to show survival results of the largest LC screening program in Spain with low dosage computed tomography (LDCT).MethodsClinical records from International Early Lung Cancer Detection Program (IELCAP) at Valencia, Spain were analysed. This program recruited volunteers, ever-smokers aged 40-80 years, since 2008. Results are compared to those from other similar sizeable programs.ResultsA total of 8278 participants were screened with at least two-rounds until November 2020. A mean of 6 annual screening rounds were performed. We detected 239 tumours along 12-year follow-up. Adenocarcinoma was the most common histology, being 61.3% at stage I. The lung cancer prevalence and incidence proportion was 1.5% and 1.4%, respectively with an annual detection rate of 0.17. One-year survival and 10-year survival were 90% and 80.1%, respectively. Adherence was 96.84%.ConclusionLargest lung cancer screening in Spain shows that survival is improved when is performed in multidisciplinary team experienced in management of LC, and is comparable to similar screening programs.     

基于肿瘤浸润深度及大小的TTS 评分系统对可切除胃癌患者预后判断的价值

目的:探讨基于肿瘤浸润深度及大小的TTS评分系统对可切除胃癌患者预后判断的价值。方法:选择行根治性切除的234例胃癌患者为研究对象。根据肿瘤浸润深度（T分期）和肿瘤大小构建TTS评分系统，分为TTS 0级、1级、2级，代表肿瘤侵袭性逐渐升高。通过Cox多因素模型分析TTS评分系统作为预后判断工具的可行性。结果:T1-T4期患者肿瘤的平均大小分别为(3.3±2.7)cm、(4.1±3.1)cm、(6.6±3.1)cm、(9.4±4.9)cm。单因素分析显示，肿瘤浸润深度与肿瘤大小、淋巴结转移、TNM分期、淋巴浸润和血管浸润显著相关(P＜0.01)。ROC曲线显示，45 mm为肿瘤大小的最佳界值，可有效区分患者是否存在淋巴结转移，曲线下面积(area under curve,AUC)为0.762。综合肿瘤大小的临界值和肿瘤浸润深度，构建TTS评分系统,TTS 0级、TTS 1级和TTS 2级的患者5年生存率分别为95.6%、83.3%和70.2%，两两比较后发现，不同TTS状态患者的生存率之间均存在显著差异(P＜0.01)。Cox多因素分析发现，TTS评分是影响患者预后的独立性危险因素（P＜0.05）。结论:本研究根据肿瘤浸润深度和大小构建了TTS评分系统，并证明了TTS评分与胃癌患者的预后密切相关。     

Are We Overtreating Patients With T1a HER2+ Breast Cancer? An Analysis of the National Cancer Database

IntroductionThe potential benefit of systemic therapy in patients with T1a HER2+ cancers is not well understood, and no consensus guidelines exist. We sought to investigate practice patterns of chemotherapy use in this population.MethodsFrom the National Cancer Database (2013-2018), we identified female patients with HER2+ cancers staged as cT1aN0 or pT1aN0 and stratified by receipt of chemotherapy. Using univariate and multivariable analyses we assessed the clinicopathologic features associated with the receipt of chemotherapy. We also compared rates of overall survival (OS).ResultsOf 5176 women with cT1aN0 HER2+ cancers, 88 (2%) received neoadjuvant chemotherapy. Younger age and hormone-receptor (HR) negative tumors were factors independently associated with receipt of neoadjuvant chemotherapy (all P < .001). Of 11,688 women with pT1aN0 HER2+ cancers, 5,588 (48%) received adjuvant chemotherapy. Rates of use increased over the analysis period from 39% in 2013 to 53% in 2018 (P < .001). Factors independently associated with receipt of adjuvant chemotherapy included younger age, having a poorly differentiated tumor, exhibiting lymphovascular invasion, undergoing adjuvant radiation (all P < .001). There were no differences in OS when comparing those who did and did not receive chemotherapy in either group.ConclusionsThe use of chemotherapy in patients with HER2+ T1a cancers is increasing over time and is, as expected, more common among patients with unfavorable clinicopathologic features. Since no prognostic algorithm currently exists, more prospective data is needed to understand which of these patients may derive benefit from systemic therapy and which may safely avoid the morbidity of chemotherapy.     

Biomolecular alterations in the early life stages of four food fish following acute exposure of Triclosan

We investigated the effect of acute concentrations of triclosan (TCS; 96 h exposure and 10d post exposure) on the free amino acid, primary (SDS-PAGE) and secondary (FT-IR) structure of proteins in the embryos/larvae of Cyprinus carpio, Ctenopharyngodon idella, Labeo rohita and Cirrhinus mrigala. A concentration dependent increase in free amino acids, upregulation of polypeptides (100 and 70 kDa in C. carpio, C. idella and L. rohita, 55, 45, 36 kda in C. idella and L. rohita and 22 kDa in all the fish) and a decline in percent area of all the selected peaks of the FT-IR spectra was observed after exposure and recovery period. The decline in percent area was greatest for L. rohita at peak 1080 – 1088 cm⁻¹ (−75.99%) after exposure and at peak 2854 – 2855 cm⁻¹ (−53.59%) after recovery. Curve fitting analysis revealed a decrease in α-helices and increase in β-sheets in all fish after exposure and recovery period. The results suggest that TCS elicits alterations in biomolecules of fish embryos.     

全身型幼年特发性关节炎患儿合并巨噬细胞活化综合征早期预警量表建立的研究

目的　总结全身型幼年特发性关节炎（sJIA）合并巨噬细胞活化综合征（MAS）的早期临床特征、实验室及辅助检查特点，筛选对诊断MAS有预警作用的检测指标，并量化形成量表，以达到快速对疾病“早识别，早治疗，降低病死率”的目的。方法　回顾性分析2006年1月至2016年1月，北京儿童医院风湿免疫科收治sJIA合并MAS患儿的病例资料，在临床及辅助检查观测指标中筛选出对早期诊断MAS有评估意义的候选指标；通过ROC曲线的方法选择最佳的诊断界限值（Cut-off值）；采用多因素Logistic回归分析MAS独立危险因素，效果以优势比（OR）表示，计算95％置信区间；经过同行评议对上述因素进行权重分值量化并最终形成积分表。结果　390例sJIA患儿，其中141例合并MAS。临床表现全部患儿均有高热、肝脾和（或）淋巴结进行性增大、血液系统受累，中枢神经系统受累患儿19例。单因素Logistic回归及ROC曲线分析结果显示，MAS组和重症sJIA组比较，10个变量存在显著差异（P<0.05）。Logistic回归分析结果显示，Fib ≤3.11g/L、WBC≤11.0×109/L、SF/ESR≥99.4及PLT≤260×109/L为MAS的独立危险因素。绘制模型ROC曲线，灵敏度和特异度分别为92.42%、81.20%。经过30位同行专家评议并投票，分别对上述患儿进行MAS诊断，并对各项临床表现及检验检查项目进行权重值评分，形成早期预警量表。结论　临床表现结合辅助检查结果，利用积分量表的形式，快速判断重症sJIA是否合并早期MAS，对诊断窗口提前，提高MAS诊断率，降低漏诊及前瞻性研究提供参考。     

早期左侧乳腺癌保乳术后大分割放疗同期瘤床加量的安全性及心脏亚结构剂量评估的重要性研究

背景与目的： 早期乳腺癌保乳术后辅助大分割放疗已被指南推荐,但大分割放疗同期瘤床加量是否可行目前尚无定论。本随机对照研究对大分割同期瘤床加量和常规分割同期瘤床加量两种放疗计划的心脏剂量学参数和心超指标等进行比较,以评估前者在心脏毒性方面的安全性。方法： 纳入复旦大学附属中山医院2017年3月—2018年3月收治的符合入组标准的早期乳腺癌保乳术后左侧乳腺癌患者40例,随机分为两组,20例制定大分割同期瘤床加量放疗计划（全乳放疗剂量4 005 cGy/15次,同期瘤床加量至4 500 cGy/15次）,20例制定常规分割同期瘤床加量放疗计划（全乳放疗剂量5 000 cGy/25次,同期瘤床加量至6 000 cGy/25次）,比较两组患者的心脏剂量学参数和心脏超声检查指标,剂量学参数包括全心脏、左心室、右心室、右室游离壁、室间隔和冠状动脉左前降支（left anterior descending,LAD）的平均剂量（D_mean）以及各结构受到5~35 Gy照射的相对体积分数（V_{5 Gy}~V_{35 Gy}）。心超随访指标包括主动脉根部直径、左房内径、左室舒张末内径、左室收缩末内径、室间隔厚度、左室后壁厚度、肺动脉收缩压、左室射血分数、E峰、A峰、E峰减速时间（E-peak deceleration time,DT）、E、A、E/A和S波峰值。此外,对两组放疗后乳房的美容效果也进行评估。结果： 大分割组和常规分割组全心脏的D_mean分别为（471.86±170.54）和（733.07±79.11）cGy（P<0.05）,全心脏的V_{20 Gy}为（3.63±1.74）%和（8.43±0.74）%（P <0.05）,V_{30 Gy}为（1.55±1.15）%和（4.48±1.01）%（P<0.05）,LAD的D_mean分别为（1 250.17±600.33）和（1 847.20±933.23）cGy（P>0.05）,左心室的D_mean分别（908.64±865.60）和（946.93±116.13）cGy（P>0.05）,右心室的D_mean分别为（590.37±197.99）和（905.73±180.82）cGy（P<0.05）,右室游离壁的D_mean分别为（939.40±284.23）和（1 597.30±446.31）cGy（P <0.05）,室间隔的D_mean分别为（637.49±248.19）和（988.60±159.77）cGy（P<0.05）。随访1年,大分割组与常规分割组相比,心超指标均在正常区间,差异均无统计学意义（P>0.05）。两组美容效果也无显著差异（P>0.05）。结论： 大分割同期瘤床加量放疗计划较常规分割计划可以显著降低全心脏和部分心脏亚结构的受照剂量,随访1年心超指标无异常,临床采用本研究所用的剂量分割方式是安全可行的。仅限制全心脏剂量无法有效保护LAD,建议勾画心脏亚结构并单独限制剂量以更好地保护心脏。     

Application of metagenomic next-generation sequencing for suspected infected pancreatic necrosis

BackgroundMetagenomic next-generation sequencing (mNGS) is increasingly used for the clinical diagnosis of infectious diseases, but there is a paucity of data regarding the application of mNGS in the early diagnosis of infected pancreatic necrosis (IPN).ObjectiveTo investigate the clinical application value of mNGS in the pathogenic diagnosis of IPN.MethodsForty-two patients with suspected IPN were prospectively and consecutively enrolled from August 2019 to August 2021. Blood samples were collected for mNGS and microbial culture simultaneously during fever (T ≥ 38.5 °C). For patients who had indications of surgical interventions, peri-pancreatic specimens were collected for mNGS and microbial culture simultaneously during the first surgical intervention to confirm IPN. The clinical performance of mNGS and microbial culture were compared.ResultsA total of 21 patients (50.0%) were confirmed to have IPN during hospitalization. The sensitivity of blood mNGS was significantly higher than blood culture (95.2% vs. 23.8%, P < 0.001) in diagnosing IPN. The negative predictive value of blood mNGS was 90.0%. The turnaround time of mNGS was significantly shorter than that of microbial culture [(37.70 ± 1.44) vs. (115.23 ± 8.79) h, P < 0.01] and the average costs of mNGS accounted for 1.7% of the average total cost of hospitalization. The survival analysis demonstrates that the positive blood mNGS result was not associated with increased mortality (P = 0.119).ConclusionsWith more valuable diagnostic performance and shorter turnaround time, clinical mNGS represents a potential step forward in the early diagnosis of IPN.