Volume 134 · Issue 17 · 2021

有大量证据支持肺康复作为慢性呼吸系统疾病患者综合管理的一个安全且具有成本效益的组成部分的作用。^[1]尽管有广泛的全球指南支持肺康复转诊，但美国最近的一项研究显示，只有约1.2%的慢性阻塞性肺疾病（COPD）患者能够获得肺康复服务。^[2]作者引用的肺康复参与率低的原因是转诊系统不充分。在中国，肺康复处于起步阶段，与发达国家相比，转诊到结构化肺康复计划不太常见。

With the ongoing worldwide coronavirus disease 2019 (COVID-19) pandemic, an increasing number of viral variants are being identified, which poses a challenge for nucleic acid-based diagnostic tests. Rapid tests, such as real-time reverse transcription-polymerase chain reaction (rRT-PCR), play an important role in monitoring COVID-19 infection and controlling its spread. However, the changes in the genotypes of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants may result in decreased sensitivity of the rRT-PCR assay and it is necessary to monitor the mutations in primers and probes of SARS-CoV-2 detection over time.

We developed two rRT-PCR assays to detect the RNA-dependent RNA polymerase (RdRp) and nucleocapsid (N) genes of SARS-CoV-2. We evaluated these assays together with our previously published assays targeting the ORF1ab and N genes for the detection and confirmation of SARS-CoV-2 and its variants of concern (VOCs). In addition, we also developed two rRT-PCR assays (S484K and S501Y) targeting the spike gene, which when combined with the open reading frames (ORF)1ab assay, respectively, to form duplex rRT-PCR assays, were able to detect SARS-CoV-2 VOCs (lineages B.1.351 and B.1.1.7).

Using a SARS-CoV-2 stock with predetermined genomic copies as a standard, the detection limit of both assays targeting RdRp and N was five copies/reaction. Furthermore, no cross-reactions with six others human CoVs (229E, OC43, NL63, HKU1, severe acute respiratory syndrome coronavirus and Middle East respiratory syndrome coronavirus) were observed using these assays. In addition, the S484K and S501Y assays were combined with the ORF1ab assay, respectively.

Four rRT-PCR assays (RdRp, N, S484K, and S501Y) were used to detect SARS-CoV-2 variants, and these assays were shown to be effective in screening for multiple virus strains.

The Nuclear Dbf2-related (NDR1) kinase is a member of the NDR/LATS family, which was a supplementary of Hippo pathway. However, whether NDR1 could inhibit glioblastoma (GBM) growth by phosphorylating Yes-associated protein (YAP) remains unknown. Meanwhile, the role of NDR1 in GBM was not clear. This study aimed to investigate the role of NDR1-YAP pathway in GBM.

Bioinformation analysis and immunohistochemistry (IHC) were performed to identify the expression of NDR1 in GBM. The effect of NDR1 on cell proliferation and cell cycle was analyzed utilizing CCK-8, clone formation, immunofluorescence and flow cytometry, respectively. In addition, the xenograft tumor model was established as well. Protein interaction was examined by Co-immunoprecipitation and immunofluorescence to observe co-localization.

Bioinformation analysis and IHC of our patients’ tumor tissues showed that expression of NDR1 in tumor tissue was relatively lower than that in normal tissues and was positively related to a lower survival rate. NDR1 could markedly reduce the proliferation and colony formation of U87 and U251. Furthermore, the results of flow cytometry showed that NDR1 led to cell cycle arrest at the G1 phase. Tumor growth was also inhibited in xenograft nude mouse models in NDR1-overexpression group. Western blotting and immunofluorescence showed that NDR1 could integrate with and phosphorylate YAP at S127 site. Meanwhile, NDR1 could mediate apoptosis process.

In summary, our findings point out that NDR1 functions as a tumor suppressor in GBM. NDR1 is identified as a novel regulator of YAP, which gives us an in-depth comprehension of the Hippo signaling pathway.

The mortality rate among patients with nasopharyngeal carcinoma (NPC) has improved significantly with the advent of chemoradiotherapy strategies. However, distant metastasis remains problematic. Tumor-specific reactivity in cancer patients has been detected exclusively in CD39+ T cells, particularly in CD39+CD103+ T cells. Circulating cancer-specific T cells are important for protecting against metastasis. This study aimed to evaluate the predictive value of circulating CD39+CD8+ T cells for metastasis in patients with NPC.

We performed a cross-sectional, longitudinal study of 55 patients with newly diagnosed NPC of stage III-IVa. All patients were initially treated with standard combined chemoradiotherapy. Blood samples were obtained from 24 patients before and at 1 month and 6 months after treatment. T cell expression of CD39 and CD103, together with the markers of T cell exhaustion programmed death-1 (PD-1)/T cell immunoglobulin and mucin domain-containing protein 3 (Tim-3) and markers of cell differentiation CD27/CC-chemokine receptor 7/CD45RA, was examined by flow cytometry. The Wilcoxon rank-sum test analysis was used to analyze the differences between two groups. Kaplan-Meier analysis was used for analysis of progression-free survival (PFS).

The expression of circulating CD39+CD8+ and CD39+CD103+ CD8+ T cells was significantly higher in patients without distant metastasis (CD39+CD8+: 6.52% [1.24%, 12.58%] vs. 2.41% [0.58%, 5.31%], Z=-2.073, P=0.038 and CD39+CD103+ CD8+: 0.72% [0.26%, 2.05%] vs. 0.26% [0.12%, 0.64%], Z=-2.313, P = 0.021). Most CD39+ T cells did not express PD-1 or Tim-3. Patients with high expression of CD39+CD103+CD8+ T cells had better PFS than patients with low expression (log rank value = 4.854, P = 0.028). CD39+CD8+ T cells were significantly elevated at 1-month post-treatment (10.02% [0.98%, 17.42%] vs. 5.91% [0.61%, 10.23%], Z = -2.943, P = 0.003). The percentage of advanced differentiated CD8+ T cells also increased at 1-month post-treatment compared with pre-treatment (33.10% [21.60%, 43.05%] vs. 21.00% [11.65%, 43.00%], Z =-2.155, P = 0.031). There was a significant correlation between elevated CD39+CD8+ T cells and increased effector memory T cells (intermediate stage: r = 0.469, P = 0.031; advanced stage: r = 0.508, P = 0.019).

CD39+CD8+ circulating T cells have preserved effector function, contributing to an improved prognosis and a reduced risk of metastasis among NPC patients. These cells may thus be a useful predictive marker for a better prognosis in patients with NPC.

Whether there is an association between serum uric acid (SUA) level and risk of mortality in the general population remains unclear. Based on the China National Survey of Chronic Kidney Disease linked to mortality data, a population-based cohort study was performed to investigate the association between SUA level and all-cause mortality, cardiovascular disease (CVD) mortality, and cancer mortality in China.

The survival status of participants in the cross-sectional survey was identified from January 1, 2006 to December 31, 2017. Only 33,268 individuals with complete SUA data among the 47,204 participants were included in the analysis. We determined the rates of all-cause mortality, CVD mortality, and cancer mortality. We used Cox proportional hazards regression models to evaluate the effect of the SUA level on mortality.

During a total of 297,538.4 person-years of follow-up, 1282 deaths occurred. In the Cox proportional hazards regression model, the rate of all-cause mortality, CVD mortality, and cancer mortality had a U-shaped association with SUA levels only in men, whereas no significant associations were detected in women. For all-cause mortality in men, the multivariable-adjusted hazard ratios (HRs) in the first, second, and fourth quartiles compared with the third quartile were 1.31 (95% confidence interval [CI] 1.04-1.67), 1.17 (95% CI 0.92-1.47), and 1.55 (95% CI 1.24-1.93), respectively. For CVD mortality, the corresponding HRs were 1.47 (95% CI 1.00-2.18), 1.17 (95% CI 0.79-1.75), and 1.67 (95% CI 1.16-2.43), respectively. For the cancer mortality rate, only a marginally significant association was detected in the fourth quartile compared with the third quartile with an HR of 1.43 (95% CI 0.99-2.08).

The association between SUA and mortality differed by sex. We demonstrated a U-shaped association with SUA levels for all-cause and CVD mortalities among men in China.

High-frequency irreversible electroporation (H-FIRE) is a novel, next-generation nanoknife technology with the advantage of relieving irreversible electroporation (IRE)-induced muscle contractions. However, the difference between IRE and H-FIRE with distinct ablation parameters was not clearly defined. This study aimed to compare the efficacy of the two treatments in vivo.

Ten Bama miniature swine were divided into two group: five in the 1-day group and five in the 7-day group. The efficacy of IRE and H-FIRE ablation was compared by volume transfer constant (Krans), rate constant (Kep) and extravascular extracellular volume fraction (Ve) value of dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI), size of the ablation zone, and histologic analysis. Each animal underwent the IRE and H-FIRE. Temperatures of the electrodes were measured during ablation. DCE-MRI images were obtained 1, 4, and 7 days after ablation in the 7-day group. All animals in the two groups were euthanized 1 day or 7 days after ablation, and subsequently, IRE and H-FIRE treated liver tissues were collected for histological examination. Student’s t test or Mann-Whitney U test was applied for comparing any two groups. One-way analysis of variance (ANOVA) test and Welch’s ANOVA test followed by Holm-Sidak’s multiple comparisons test, one-way ANOVA with repeated measures followed by Bonferroni test, or Kruskal-Wallis H test followed by Dunn’s multiple comparison test was used for multiple group comparisons and post hoc analyses. Pearson correlation coefficient test was conducted to analyze the relationship between two variables.

Higher Ve was seen in IRE zone than in H-FIRE zone (0.14 ± 0.02 vs. 0.08 ± 0.05, t= 2.408, P = 0.043) on day 4, but no significant difference was seen in Ktrans or Kep between IRE and H-FIRE zones at all time points (all P > 0.05). For IRE zone, the greatest Ktrans was seen on day 7, which was significantly higher than that on day 1 (P = 0.033). The ablation zone size of H-FIRE was significantly larger than IRE 1 day (4.74 ± 0.88 cm²vs. 3.20 ± 0.77 cm², t = 3.241, P = 0.009) and 4 days (2.22 ± 0.83 cm²vs. 1.30 ± 0.50 cm², t = 2.343, P = 0.041) after treatment. Apoptotic index (0.05 ± 0.02 vs. 0.73 ± 0.06 vs. 0.68 ± 0.07, F = 241.300, P < 0.001) and heat shock protein 70 (HSP70) (0.03 ± 0.01 vs. 0.46 ± 0.09 vs. and 0.42 ± 0.07, F= 64.490, P < 0.001) were significantly different between the untreated, IRE and H-FIRE zones, but no significant difference was seen in apoptotic index or HSP70 between IRE and H-FIRE zone (both P > 0.05). Electrode temperature variations were not significantly different between the two zones (18.00 ± 3.77°C vs. 16.20 ± 7.45°C, t = 0.682, P = 0.504). The Ktrans value (r = 0.940, P = 0.017) and the Kep value (r = 0.895, P = 0.040) of the H-FIRE zone were positively correlated with the number of hepatocytes in the ablation zone.

H-FIRE showed a comparable ablation effect to IRE. DCE-MRI has the potential to monitor the changes of H-FIRE ablation zone.

Long non-coding RNA (lncRNA) actin filament-associated protein 1 antisense RNA 1 (AFAP1-AS1) functions as a competing endogenous RNA to regulate target genes expression by sponging microRNAs (miRs) to play cancer-promoting roles in cancer stem cells. However, the regulatory mechanism of AFAP1-AS1 in cervical cancer (CC) stem cells is unknown. The present study aimed to provide a new therapeutic target for the clinical treatment of CC.

Hyaluronic acid receptor cluster of differentiation 44 variant exon 6 (CD44v6)(+) CC cells were isolated by flow cytometry (FCM). Small interfering RNAs of AFAP1-AS1 (siAFAP1-AS1) were transfected into the (CD44v6)(+) cells. The levels of AFAP1-AS1 were measured by quantitative real-time PCR (qRT-PCR). Sphere formation assay, cell cycle analysis, and Western blotting were used to detect the effect of siAFAP1-AS1. RNA pull-down and luciferase reporter assay were used to verify the relationship between miR-27b-3p and AFAP1-AS1 or vascular endothelial growth factor (VEGF)-C.

CD44v6(+) CC cells had remarkable stemness and a high level of AFAP1-AS1. However, AFAP1-AS1 knockdown with siAFAP1-AS1 suppressed the cell cycle transition of G(1)/S phase and inhibited self-renewal of CD44v6(+) CC cells, the levels of the stemness markers octamer-binding transcription factor 4 (OCT4), osteopontin (OPN), and cluster of differentiation 133 (CD133), and the epithelial-mesenchymal transition (EMT)-related proteins Twist1, matrix metalloprotease (MMP)-9, and VEGF-C. In the mechanism study, miR-27b-3p/VEGF-C signaling was demonstrated to be a key downstream of AFAP1-AS1 in the CD44v6(+) CC cells.

LncRNA AFAP1-AS1 knockdown inhibits the CC cell stemness by upregulating miR-27b-3p to suppress VEGF-C.

Endometrial cancer (EC) has been one of the most general cancers with respect to gynecological malignancies; however, there are debates on clinical strategies concerning treatments especially for patients with grade 3 (G3) endometroid endometrial cancer (EEC). Present study aimed to evaluate the lymphatic metastasis (LM) related factors and figure out the necessity of lymphadenectomy for G3 EEC patients.

From January 2009 to April 2019, 3751 EC patients were admitted to Obstetrics and Gynecology Hospital of Fudan University. Clinical characteristics include age, grade, stage, and clinical pathological features. A total of 1235 EEC patients were involved in the multivariable analysis. Three hundred and eighty-one patients were involved in the survival analysis and the data attributed to sufficient follow-up information. Kaplan-Meier curve and log-rank test were utilized to analyze the survival rate.

Among the 1235 EEC patients, 181 (14.7%) were categorized as G3 and 1054 (85.3%) were grade 1 to grade 2 (G1-2). Multivariate analysis demonstrated that lymphovascular space invasion, adnexal involvement, and cervical stroma involvement were independent risk factors of LM in G3 cohort with odds ratio 3.4, 5.8, and 8.9; 95% confidence interval 1.1-10.6, 1.5-22.4, and 2.8-28.0, respectively. LM rates increased from 3.3% (3/92) to 75% (9/12) for G3 EEC cohort as related factor numbers increased from one to three. There were no differences between G3 and G1-2 EEC in overall survival and progression free survival. Additionally, no survival advantage was observed for G3 EEC patients at early stage with different plans of adjuvant treatment.

For G3 EEC patients without other pathological positive factor, the LM rate is lower than those with other pathological positive factor. Survival analysis showed no difference between G3 cohort and G1-2 cohort. Also, different adjuvant treatments had no impact on the overall survival for G3 EEC patients.

冠状病毒疾病2019（新冠肺炎）在全球范围内造成了巨大的人类生命损失。住院患者的气管插管率估计为2.3%。^[1]作为一种产生气溶胶的程序，插管可能会增加医护人员感染新冠肺炎的风险。避免气溶胶的产生和改善插管过程中的氧合有时是相互冲突的，这使得新冠肺炎患者的插管不同于其他急性呼吸窘迫综合征（ARDS）患者的插管。我们进行了一项调查，以收集新冠肺炎患者预氧合、诱导和插管程序的信息。我们的研究旨在比较曾对新冠肺炎患者进行气管插管的医生与从未对新冠肺炎患者进行气管插管的医生之间插管性能的差异。

前交通动脉（ACoA）动脉瘤在颅内动脉瘤中所占比例最大，最容易破裂。^[1]主要治疗方法为血管内栓塞和传统开颅夹闭术。^[2]近二十年来，随着介入材料和技术的飞速发展，许多医疗中心已将血管内栓塞作为颅内动脉瘤的首选治疗方法。然而，一些动脉瘤仍然需要传统的开颅夹闭术。经蝶内镜技术近年来也发展迅速。^[3]由于颅内动脉瘤多在Willis环内，发生在颅底腹侧，少数神经外科医生尝试使用鼻内镜入路（EEA）进行夹闭。^[4,5,6]然而，文献报道的病例数量较少，需要更多的解剖学研究来深入分析该方法和技术。

哮喘被认为是以慢性气道炎症和重塑为特征的免疫失调。越来越多的证据表明，哮喘表现出各种炎症特征和细胞类型。最近，调节性T细胞（Tregs）和T辅助细胞（Th）17细胞被描述为与Th1和Th2细胞不同的两个亚群，并且Treg/Th17失衡与哮喘的发展相关。

致编辑：培美曲塞加铂（顺铂或卡铂）化疗双联疗法被广泛推荐作为未发现基因突变的非小细胞肺癌（NSCLC）的标准一线治疗。^[1]然而，它们的有效性已得到证实，中位生存期约为10.2至13.0个月，5年生存率为13%至15%。培美曲塞初始化疗已被证实是晚期肺腺癌的有效方案，但没有预测培美曲塞疗效获益的特定遗传生物标志物。^[2]以培美曲塞为基础的方案治疗的患者也可以在诱导化疗后接受单药作为维持治疗。此外，培美曲塞在既往研究中显示出优异的患者耐受性和安全性特征。先前的报告表明，在非小细胞肺癌患者的特定亚组中，使用或不使用维持方案治疗酪氨酸激酶抑制剂获得性耐药的培美曲塞再激发可能是一种可行的选择。一种可能的解释是培美曲塞敏感性肿瘤细胞克隆在培美曲塞再激发后仍可存活。由于晚期非小细胞肺癌缺乏标准的多线治疗，我们假设培美曲塞单药或联合方案再激发可获益。本研究重点关注以培美曲塞为基础的再激发方案在局部晚期或转移性肺腺癌患者中的疗效和安全性，这些患者已从一线培美曲塞铂类化疗中获得有益缓解。

致编辑：在中国2019冠状病毒病（COVID-19）疫情期间，尸体器官捐赠急剧减少，这加剧了肺移植（LTx）患者的等候名单发病率和死亡率。我们报告了一个病例，其中我们能够通过使用供体的左肺为一个受体进行单个左侧LTx来最大限度地利用来自单个尸体供体的肺，并且通过将供体的右肺进行二分，我们能够为另一个受体进行双侧肺叶移植。

致编辑：高度恶性的肺腺癌（LUAD）是肺癌最常见的组织学亚型，也是世界上癌症相关死亡的主要原因。^[1]肿瘤免疫微环境主要由浸润性免疫和基质细胞组成，对LUAD患者的预后有重要影响。^[2]近年来，高通量测序技术的突飞猛进深刻改变了我们对肿瘤研究的认识。它利用大量公开的临床数据，帮助研究人员更有效地研究肿瘤的特性，提高我们诊断、治疗和预防癌症的能力。本研究的目的是使用表达数据（ESTIMATE）算法估计恶性肿瘤组织中的基质和免疫细胞，从癌症基因组图谱（TCGA）数据库中计算LUAD患者的免疫和基质评分，探索评分与LUAD患者生存之间的相关性，并根据免疫/基质评分识别差异表达基因（DEGs）。此外，我们还判断了DEGs的预后价值，并进一步研究了其潜在的分子功能。

致编辑：原发性肺癌是最常诊断的恶性肿瘤类型，是全球癌症死亡的主要原因。非小细胞肺癌(NSCLC)占原发性肺癌总发病率的80%至85%，并分为两种不同的组织学亚型：肺腺癌(LUAD)和肺鳞状细胞癌(LUSC)。LUAD占所有原发性肺癌病例的近30%至35%，并且在大多数国家其复发似乎正在增加。^[1]作为一种普遍存在的mRNA内部修饰，N6-甲基腺苷（m6A）甲基化修饰对于触发癌症的发展具有重要意义，可作为多种癌症的促癌因子。^[2,3]在这项研究中，我们旨在为LUAD患者开发一种以m6A RNA甲基化调节因子为前提的预后信号。

致编辑：2019冠状病毒病（COVID-19）已成为全球大流行。为了最大限度地降低病毒在社区传播的风险，一些国家要求对相隔≥24小时采集的鼻咽拭子进行两次严重急性呼吸综合征冠状病毒2型（新型冠状病毒）RNA逆转录聚合酶链反应（RT-PCR）阴性结果，以便出院和停止隔离。因此，了解病毒清除的时间至关重要。然而，报告新冠肺炎患者病毒清除时间及其危险因素的数据有限。^[1,2]此外，这些研究中患者数量有限，病毒清除的风险因素不一致。因此，我们旨在确定新冠肺炎患者咽拭子中病毒清除的时间和危险因素。

致编辑：胰腺癌（PC）的病因和发病机制知之甚少。人们早就认识到，各种相互交织的因素，包括遗传易感性和环境刺激（如香烟、糖尿病和肥胖），是PC患者高发病率和高死亡率的潜在危险因素。^[1]脂肪因子在肿瘤发病中的调节作用仍存在争议。已经普遍接受的是，肥胖相关病症增加了对包括PC在内的各种肿瘤的易感性。肥胖增加的PC患者预后较差和死亡率较高的风险增加，这表明肥胖相关因素（即瘦素和AdipoQ）在PC发病和/或发展中的意义正变得越来越突出。然而，与非PC个体相比，PC的循环瘦素和AdipoQ水平仍有争议。因此，我们试图进行荟萃分析来解决有争议的问题。

致编辑：肾细胞癌（RCC）是成人中十大主要癌症类型之一。目前的研究表明，透明细胞肾细胞癌（ccRCC）仍然是一种致命的肿瘤疾病，并与大多数癌症相关的死亡有关。^[1]及时诊断对于肿瘤患者的治疗极为重要。根据临床诊断标志物结果调整治疗方案对患者预后有积极作用。肾切除术后局部复发或远处转移的风险仍然很高。酪氨酸激酶抑制剂（TKI）在晚期ccRCC患者中显示出令人鼓舞的疗效。^[2]不幸的是，TKI耐药性仍在发展，并可导致癌症的不良进展。^[3]因此，迫切需要探索更有效的预后生物标志物并鉴定用于ccRCC治疗的新药物靶点。

致编辑：复发性妊娠丢失（RPL）被定义为妊娠24周前两次或两次以上的妊娠丢失，它影响约1%至2%的夫妇。^[1]RPL的病因包括代谢/内分泌异常、遗传因素、解剖因素、免疫紊乱、血栓形成倾向、男性因素和心理因素。^[1,2]全身性自身免疫性疾病，如抗磷脂综合征(APS)，^[3]未分化结缔组织病(UCTD)、^[4]和系统性红斑狼疮(SLE)^[5]近年来被发现是RPL的重要原因。了解不同全身性自身免疫性疾病与RPL之间的关联，以及能够提供合适的诊断和治疗具有重要意义。

在文章“淀粉样蛋白和tau阳性轻度认知障碍：痴呆进展的临床和生物标志物特征”中，该文章出现在第134卷第14期第1709页中华医学杂志，应在作者部分添加以下文字“阿尔茨海默病神经成像倡议”。完整的作者应更正为“Hong-Chun Wei¹,李冰¹, Kok Pin Ng²、清-*赋³,董胜杰⁴,毛文巴¹,闵孔⁵;阿尔茨海默病神经影像计划”。第1717页，资助部分应更正为“本研究得到山东省重点研发项目（编号：2018GSF118235）、山东省自然科学基金（编号：ZR2016HL16）和青岛大学附属烟台毓璜顶医院青年科研启动基金（编号：2020-25）的资助。”