2021年 · 第134卷第17期

There is considerable evidence supporting the role of pulmonary rehabilitation as a safe and cost-effective component of integrated management in patients with chronic respiratory illness.^[1]Despite extensive global guidelines endorsing pulmonary rehabilitation referral, a recent study in the United States revealed that only approximately 1.2% of patients with chronic obstructive pulmonary disease (COPD) have access to pulmonary rehabilitation services.^[2]The reason cited by the authors for the low participation rate in pulmonary rehabilitation was an inadequate referral system. In China, pulmonary rehabilitation is in its infancy, and referral to a structured pulmonary rehabilitation program is less common compared with the developed countries.

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.

Coronavirus disease 2019 (COVID-19) has caused a dramatic loss of human life worldwide. The rate of tracheal intubation is estimated to be 2.3% in hospitalized patients.^[1]As an aerosol-generating procedure, intubation may increase the risk of COVID-19 infection in healthcare workers. Avoiding the generation of aerosol and improving the oxygenation during intubation is sometimes conflicting, which makes the intubation in patients with COVID-19 different from that in other patients with acute respiratory distress syndrome (ARDS). We conducted a survey to collect information on preoxygenation, induction, and intubation procedures in COVID-19 patients. Our study aims to compare the difference in intubation performance between doctors who have ever done tracheal intubation in COVID-19 patients versus those who have never done tracheal intubation in COVID-19 patients.

Anterior communicating artery (ACoA) aneurysms account for the largest proportion of intracranial aneurysms and are the most likely to rupture.^[1]The main treatment methods are endovascular embolization and traditional craniotomy clipping.^[2]With the rapid development of interventional materials and technologies in the last two decades, many medical centers have adopted endovascular embolization as the preferred treatment for intracranial aneurysms. However, some aneurysms still require traditional craniotomy clipping. Transsphenoidal endoscopy technology has also developed rapidly in recent years.^[3]Because intracranial aneurysms are mostly within the circle of Willis and occur on the ventral side of the skull base, a few neurosurgeons have tried to use an endoscopic endonasal approach (EEA) for clipping.^[4,5,6]However, the number of cases reported in the literature is small and more anatomical studies are needed to deeply analyze the approach and technique.

Asthma is considered dysregulated immunity featuring of chronic airway inflammation and remodeling. Mounting evidence reveals that asthmatics show various inflammatory profiles and cell types. Recently, regulatory T cells (Tregs) and T-helper (Th) 17 cells have been described as two distinct subsets from Th1 and Th2 cells and Treg/Th17 imbalance is correlated with the development of asthma.

To the Editor: Pemetrexed plus platinum (cisplatin or carboplatin) chemotherapy doublets are widely recommended as the standard first-line treatment for non-small cell lung cancer (NSCLC) with no identified genetic mutations.^[1]However, their efficiency has been confirmed with a median survival of approximately 10.2 to 13.0 months and a 5-year survival rate of 13% to 15%. Initial chemotherapy with pemetrexed has been confirmed as an effective regimen for advanced lung adenocarcinoma without specific genetic biomarker predicting the benefit of pemetrexed efficacy.^[2]Patients treated with pemetrexed-based regimens can also receive a single agent as maintenance treatment after the induction chemotherapy. In addition, pemetrexed has shown an excellent patient tolerance and safety profile in previous studies. Previous reports suggested that rechallenge with pemetrexed with or without maintenance regimens for acquired resistance of tyrosine kinase inhibitors might be a feasible option in specific subgroup of NSCLC patients. One possible explanation is that clones of pemetrexed-sensitive tumor cells may still survive following the pemetrexed rechallenge. Since the absence of standard multiline treatment in advanced NSCLC, we hypothesized a benefit from rechallenge with pemetrexed alone or combination regimens. This study focused on the efficacy and safety of rechallenge pemetrexed-based regimens in patients with locally advanced or metastatic lung adenocarcinoma, who had achieved beneficial response from first-line pemetrexed platinum chemotherapy.

To the Editor: During the coronavirus disease 2019 (COVID-19) pandemic in China, cadaveric organ donation became drastically reduced, which exacerbated the waitlist morbidity and mortality of lung transplant (LTx) patients. We report a case where we were able to maximize the utilization of the lungs from a single cadaveric donor by performing a single left LTx for one recipient using the donor’s left lung, and by bipartitioning the donor’s right lung, we were able to perform bilateral lobar transplantation for another recipient.

To the Editor: Lung adenocarcinoma (LUAD) with a high degree of malignancy is the most common histological subtype of lung cancer, and the leading cause of cancer-related death in the world.^[1]Tumor immune microenvironment is mainly composed of infiltrating immune and stromal cells, which have an important influence on the prognosis of LUAD patients.^[2]In recent years, rapid advances in high-throughput sequencing technology have profoundly changed our understanding of tumor research. It uses a large amount of public clinical data to help researchers more effectively to study the characteristics of tumors and improve our ability to diagnose, treat, and prevent cancer. The purpose of this study was to use the estimation of stromal and immune cells in malignant tumor tissues using expression data (ESTIMATE) algorithm to calculate the immune and stromal scores of LUAD patients from The Cancer Genome Atlas (TCGA) database, explore the correlation between the scores and the survival of LUAD patients, and identify differentially expressed genes (DEGs) based on the immune/stromal scores. In addition, we also judged the prognostic value of DEGs and further studied their potential molecular functions.

To the Editor: Primary lung cancer is the most commonly diagnosed type of malignant tumor and is the leading cause of cancer death worldwide. Non-small cell lung cancer (NSCLC) makes up 80% to 85% of the overall incidents of primary lung cancer and is classified into two distinct histological subtypes: lung adenocarcinoma (LUAD) and lung squamous cell carcinoma (LUSC). LUAD constitutes nearly 30% to 35% of all primary lung cancer cases, and the recurrence of it seems to be increasing in most countries.^[1]As a prevalent mRNA internal modification, N6-methyladenosine (m6A) methylation modification is of great significance for triggering the development of cancer and can be used as a cancer-promoting factor in many cancers.^[2,3]In this study, we aim to develop an m6A RNA methylation regulators-premised prognostic signature for LUAD patients.

To the Editor: Coronavirus disease 2019 (COVID-19) has become a global pandemic. To minimize the risk of viral transmission in the community, some countries require two negative results of reverse transcription polymerase chain reaction (RT-PCR) for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA on nasopharyngeal swabs collected ≥24 h apart for hospital discharge and discontinuation of isolation. Thus, understanding the time of viral clearance is essential. However, there are limited data reporting the viral clearance time and its risk factors in COVID-19 patients.^[1,2]Moreover, the numbers of patients were limited and the risk factors of viral clearance were inconsistent in these studies. Therefore, we aim to determine the time and risk factors of viral clearance in throat swabs in COVID-19 patients.

To the Editor: The etiology and the pathogenesis of pancreatic cancer (PC) are poorly understood. It has long been recognized that various intertwined factors, including genetic susceptibility and environmental stimuli (such as cigarettes, diabetes mellitus, and adiposity), are potential risk factors for high morbidity and mortality in PC patients.^[1]The regulatory role of adipokines in the pathogenesis of tumors remains controversial. It has been generally accepted that obesity-related conditions increase susceptibility to various tumors, including PC. PC patients with increased adiposity have a heightened risk for poorer prognosis and higher mortality, which suggests that the significance of obesity-related factors (ie, leptin and AdipoQ) in the onset and/or development of PC is becoming increasingly prominent. However, circulating leptin and AdipoQ levels in PC compared with those in non-PC individuals remain controversial. Hence, we sought to perform a meta-analysis to settle the issues in dispute.

To the Editor: Renal cell carcinoma (RCC) is one of the top 10 major cancer types in adults. Current research shows that clear cell renal cell carcinoma (ccRCC) remains a deadly tumor disease and is associated with most cancer-related deaths.^[1]Timely diagnosis is extremely important for the treatment of tumor patients. Adjusting the treatment plan based on the results of clinical diagnostic markers has a positive effect on patient prognosis. There is still a high risk of local recurrence or distant metastasis after nephrectomy. Tyrosine kinase inhibitor (TKI) has shown encouraging efficacy in advanced ccRCC patients.^[2]Unfortunately, TKI resistance still develops and can lead to poor progression in cancer.^[3]Thus, there is an urgent need to explore more effective prognostic biomarkers and to identify novel drug targets for ccRCC therapies.

To the Editor: Recurrent pregnancy loss (RPL) is defined as the loss of two or more pregnancies before 24 weeks of gestation, and it affects about 1% to 2% of couples.^[1]The etiology of RPL includes metabolic/endocrinological abnormalities, genetic factors, anatomical factors, immune disorders, thrombophilia, male factors, and psychological factors.^[1,2]Systemic autoimmune diseases like antiphospholipid syndrome (APS),^[3]undifferentiated connective tissue disease (UCTD),^[4]and systemic lupus erythematosus (SLE)^[5]have been found to be important causes of RPL in recent years. Understanding the associations between different systemic autoimmune diseases and RPL, as well as being able to provide suitable diagnoses and treatments, is of great significance.

In the article "Amyloid and tau positive mild cognitive impairment: clinical and biomarker characteristics of dementia progression" , which appeared in vol.134, issue14, page 1709 ofChinese Medical Journal, the following words "for Alzheimer’s disease Neuroimaging Initiative" should be added to the author section. The full authors should be corrected as "Hong-Chun Wei¹, Bing Li¹, Kok Pin Ng², Qing-Xi Fu³, Sheng-Jie Dong⁴, Mao-Wen Ba¹, Min Kong⁵; for Alzheimer’s disease Neuroimaging Initiative" . And in page 1717, the Funding Part shoule be corrected as "The study was supported by grants from the Shandong Provincial key research and development project (No.2018GSF118235), the Shandong Provincial Natural Science Foundation (No.ZR2016HL16) and Youth Research Start-up Fund of Yantai Yuhuangding Hospital Affiliated to Qingdao University (No.2020-25)."