<b>Introduction:</b> The ninth edition of the American Joint Committee on Cancer/Union for International Cancer Control (AJCC/UICC) tumor-node-metastasis (TNM) system for patients with non-metastatic nasopharyngeal carcinoma (NPC) was based on data from endemic regions, and it required validation for application to non-endemic regions. <br><br><b>Aim:</b> The aim of this study was to compare the performance of the 8<sup>th</sup> and the 9<sup>th</sup> editions of the AJCC/UICC TNM system for patients with non-metastatic NPC from non-endemic regions. <br><br><b>Materials and methods:</b> Using the Surveillance, Epidemiology, and End Results (SEER) database, 2004-2021, we re-classified NPC patients using 8<sup>th</sup> and the 9<sup>th</sup> edition staging based on surrogate markers of advanced radiological extranodal extension (nodes described as "fixed") and compared the prediction performance of both staging editions for overall survival (OS) and cancer specific survival (CSS). <br><br><b>Results:</b> Our analysis included 4981 patients with a median follow-up of 99.0 months. Comparing the 9<sup>th</sup> version of AJCC/UICC (TNM-9) with TNM-8, only 29 (1.8%) patients with N1 and 47 (3.3%) patients with N2 were upstaged to N3 in TNM-9. In comparisons based on OS, TNM-9 did not outperform TNM-8 with regard to hazard consistency (2.2326 <i>vs</i> 1.9160), hazard discrimination (1.4092 <i>vs</i> 1.0947), sample size balance (0.3476 <i>vs</i> 0.3398), and percent variance explained (0.2704 <i>vs</i> 0.2704). The area under the receiver operating characteristic curve (AUC) values of TNM-9 and TNM-8 were 0.5299 and 0.5330 for OS (P = 0.549), respectively. The AUC values of TNM-9 and TNM-8 were 0.5332 and 0.5357 for CSS (P = 0.345), respectively. <br><br><b>Conclusions:</b> Restaging according to TNM-9 criteria involved only minor changes in the distribution of nodal (N) categories. Neither the TNM-9 nor the TNM-8 staging system demonstrated adequate overall discrimination between each stage group classification in terms of OS and CSS for patients with non-metastatic NPC from a non-endemic region.

Purpose: This study aimed to illustrate the heterogeneity of immune features in small cell lung cancer (SCLC).Methods: Immunohistochemistry (IHC) staining of CD3, CD4, CD8 and PD-L1 were performed with 55 SCLC FFPE samples from radical resections. Quantitative assessment of CD3+ tumor-infiltrated lymphocytes (TILs) to present the heterogeneity in the tumor and the stroma areas. Hotspots of TILs were evaluated to illustrate the potential relationship between TIL-density and its immune competence. Programmed death ligand-1 (PD-L1) expressed on both tumor TILs (t-TILs) and stroma TILs (s-TILs) was evaluated and quantitatively described as values of tumor positive score (TPS) and combined positive score (CPS). The clinical value of TPS and CPS were further identified according to their relationship with disease-free survival (DFS).Results: More abundant CD3+ TILs were observed in the tumor stroma than that within the parenchyma (15.02 & PLUSMN;2.25% vs. 1.58 & PLUSMN;0.35%) . The amount of CD3+ s-TILs were positively correlated with DFS. The CD3+/CD4+ subset of the TILs was found more favorable to DFS compared to the CD3+/CD8+ subset. Hotspots of CD3+ TILs were observed in tumor regions and patients with more Hotspots of CD3+ TILs have better outcomes. CPS were more reliable than TPS to describe PD-L1 expression in SCLC and it was found positively correlated with tumor size and DFS.Conclusions: The immune microenvironment of SCLC was heterogeneous. Hotspots, the amount of CD3/CD4+ TILs and the CPS value were found valuable in determine the anti-tumor immunity and predicting the clinical outcome of SCLC patients.

The present study aimed to clarify the association between macrophages, tumor neo-vessels and programmed cell death-ligand 1 (PD-L1) in the tumor microenvironment and the clinicopathological features of patients with non-small cell lung cancer (NSCLC), and to explore the prognostic factors of stromal features in NSCLC. To determine this, tissue microarrays containing samples of 92 patients with NSCLC were studied using immunohistochemistry and immunofluorescence. The quantitative data demonstrated that in tumor islets, the number of CD68+ and CD206+ tumor-associated macrophages (TAMs) was 8-348 (median, 131) and 2-220 (median, 52), respectively (P<0.001). In tumor stroma, the number of CD68+ and CD206+ TAMs was 23-412 (median, 169) and 7-358 (median, 81), respectively (P<0.001). The number of CD68+ TAMs in each location of the tumor islets and tumor stroma was significantly higher than that of CD206+ TAMs, and they were significantly correlated (P<0.0001). The quantitative density of CD105 and PD-L1 in tumor tissues was 19-368 (median, 156) and 9-493 (median, 103), respectively. Survival analysis revealed that a high density of CD68+ TAMs in tumor stroma and islets and a high density of CD206+ TAMs and PD-L1 in tumor stroma were associated with worse prognosis (both P<0.05). Collectively, the survival analysis demonstrated that the high-density group was related to a worse prognosis regardless of combined neo-vessels and PD-L1 expression with the CD68+ TAMs in tumor islets and stroma, or CD206+ TAMs in tumor islets and stroma. To the best of our knowledge, the present study was the first to provide a multi-component combined prognostic survival analysis of different types of macrophages in different regions with tumor neo-vessels and PD-L1, which demonstrated the importance of macrophages in tumor stroma.

Tumor-infiltrating lymphocytes (TILs) offer a key for morphological diagnosis of lymphoepithelioma-like carcinoma (LELC) and are the foundation of oncoimmunology. To date, no reports have found a specific risk stratification value of TILs and related it to genomic variation in LELC. Based on the stromal TILs (str-TILs) ratio, we classified 105 Epstein-Barr virus (EBV)-associated LELC cases into two subtypes: patients with ≥60% str-TILs area ratio in tumor were classified as subtype I, and otherwise as subtype II. Subtype I patients had significantly better progression-free survival (PFS) and overall survival (OS). We also explored the genomic characteristics of EBV-associated LELC within different involved organs. We performed whole-exome sequencing for 51 patients with enough tissue and analyzed the genomic characteristics of EBV-associated LELC. Overall, EBV-associated LELCs were characterized by a low somatic mutation rate and copy number variations; the enriched genetic lesions affected RTK-RAS, PI3K, and cell cycle pathways. Moreover, EBV-associated LELCs from different organs were more similar to each other genetically as compared with other traditional carcinomas of the same sites-as evidenced by unsupervised clustering based on the quantitative data from both mutation signature and chromosomal aneuploidies. Notably, EBV-associated LELC patients with oncogenic driver alterations showed a worse prognosis compared with patients without such alterations. © 2022 The Pathological Society of Great Britain and Ireland.

Programmed cell death ligand 1 (PD-L1) is a critical biomarker for predicting the response to immunotherapy. However, traditional quantitative evaluation of PD-L1 expression using immunohistochemistry staining remains challenging for pathologists. Here we developed a deep learning (DL)-based artificial intelligence (AI) model to automatically analyze the immunohistochemical expression of PD-L1 in lung cancer patients. A total of 1,288 patients with lung cancer were included in the study. The diagnostic ability of three different AI models (M1, M2, and M3) was assessed in both PD-L1 (22C3) and PD-L1 (SP263) assays. M2 and M3 showed improved performance in the evaluation of PD-L1 expression in the PD-L1 (22C3) assay, especially at 1% cutoff. Highly accurate performance in the PD-L1 (SP263) was also achieved, with accuracy and specificity of 96.4 and 96.8% in both M2 and M3, respectively. Moreover, the diagnostic results of these three AI-assisted models were highly consistent with those from the pathologist. Similar performances of M1, M2, and M3 in the 22C3 dataset were also obtained in lung adenocarcinoma and lung squamous cell carcinoma in both sampling methods. In conclusion, these results suggest that AI-assisted diagnostic models in PD-L1 expression are a promising tool for improving the efficiency of clinical pathologists.

Previous small-size studies reported BRAF-mutated NSCLC patients have comparable sensitivity to immune checkpoint inhibitors (ICIs). However, how BRAF mutation affects the tumor immune microenvironment (TIME) is unknown. We performed Nanostring-panel RNA sequencing to evaluate TIME in 57 BRAF mutated and wild-type (WT) NSCLC specimens (cohort A). The efficacy of ICI monotherapy or combined therapies was determined in 417 patients with WT and BRAF mutated NSCLC (cohort B). We found that BRAF-mutant tumors had similar ratios of CD8+ T cells to Tregs, the balance of cytotoxicity gene expression signatures and immune suppressive features, and similar ICI-response-related biomarkers to WT NSCLC. A similar TIME pattern was observed between the BRAF V600E and Non-V600E subgroups of NSCLC. The further retrospective study confirmed that treatment with ICI monotherapy or combined therapies resulted in similar overall survival (OS) (HR: 0.85; 95% CI, 0.56 to 1.30; p = 0.47) and progress-free survival (PFS) (HR: 1.02; 95% CI, 0.72 to 1.44; p = 0.91) of patients with WT (n = 358) and BRAF mutant (n = 59) NSCLC. Similarly, both patients with BRAF V600E or Non-V600E NSCLC had similar responses to immunotherapy. Our findings support that BRAF mutation did not modulate TIME in NSCLC and therapeutic responses to ICIs. Patients with NSCLC harboring BRAF mutation should not be denied treatment with ICIs.

Abstract Purpose To evaluate the role of programmed death-ligand 1 (PD-L1) and mammalian target of rapamycin (mTOR) signaling pathway in locally advanced rectal cancer (LARC). Methods Between February 2012 and February 2018, 103 patients with LARC treated by neoadjuvant chemoradiotherapy (neoCRT) and total mesorectal excision (TME) were included. PD-L1, mTOR and p-mTOR of pair-matched pre-neoCRT biopsies and post-neoCRT surgical tissue were evaluated by immunohistochemistry. Results The mean combined positive score (CPS), tumor proportion score (TPS) and immune cell score (IC) of pre-neoCRT were 2.24 (0–70), 1.87 (0–70) and 0.67 (0–10), respectively. The mean CPS, TPS and IC of post-neoCRT were 2.19 (0–80), 1.38 (0–80) and 1.60 (0–20), respectively. Significant difference was observed in terms of IC between pre-neoCRT and post-neoCRT (p = 0.010). The 5-year disease-free survival (DFS) rate of the whole group was 62.4%. Multivariate analysis by Cox model indicated that pre-neoCRT TPS [hazard ratio (HR) 1.052, 95% confidence interval (CI) 1.020–1.086, p = 0.001] and post-neoCRT CPS (HR 0.733, 95% CI 0.555–0.967, p = 0.028) were associated with DFS. In the 89 patients without pathological complete response, p-mTOR and IC were upregulated after neoCRT. Conclusions For patients with LARC treated by neoCRT and TME, p-mTOR and IC were upregulated after neoCRT. Pre-neoCRT TPS and post-neoCRT CPS were independent prognostic predictors of DFS.

Background:According to the latest the World Health Organization (WHO) classification in 2015, invasive mucinous adenocarcinoma (IMA) is defined as a new pathological subtype of lung adenocarcinoma (LUAD). However, whether this rare subtype of lung pathology has any difference in prognosis than conventional LUAD is debatable. Our study attempted to compare clinical characteristics and prognosis of IMA vs. noninvasive mucinous adenocarcinomas (NMA).Methods:A total of 1,857 patients with LUAD who underwent radical resection were screened from 2010 to 2015 at Zhejiang Cancer Hospital. Patients with pulmonary IMA were matched 1:1 by using propensity scores with LUAD adjusted for clinicopathological characteristics. After follow-up, overall survival (OS) and disease-free survival (DFS) were explored by Kaplan-Meier and Cox regression analyses. Forest plots were used for subgroup analyses.Results:Following screening, 499 patients with LUAD were enrolled, with 97 IMA and 402 NMA. Compared to NMA of the lung, IMA was proportionately lower in women (50.5% vs. 63.4%; P=0.026) and nonsmokers (P<0.001). IMA was also associated with earlier tumor stage I (68.0% vs. 55.5%; P=0.033) and lower frequency of upper lobe tumors compared to NMA (P=0.007). Following propensity score matching, 97 pairs were selected, among which we found that patients with pulmonary IMA had a longer OS than those with NMA (P=0.014). According to the subgroup analysis, improved OS in the IMA cohort versus the NMA cohort was observed across various factors, including the absence of lymphovascular invasion or perineural invasion.Conclusions:In this study, we found that resectable IMA patients had a better OS than NMA patients. This study contributes to the understanding of IMA in depth, but it needs to be validated through additional multicenter studies.

Brain metastasis (BM) is a common malignant event in lung cancer. Here, we recruited 33 lung cancer patients with brain oligo-metastasis to explore the genomic features and tumor immune microenvironment (TIME) of the lung and BM independently. For genomic profiling, targeted sequencing was performed. We found that high-frequent ZFHX3 occurred in the lung (40%) and brain tumor (28%), which might relate to brain metastasis event; the vast majority of patients had lesions-shared mutations in primary tumor and BM, confirming the common clonal events; and EGFR was the most frequently clonal gene in both lung and BM, indicating its driver capability. To characterize TIME status, we also sequenced the T cell receptor (TCR) repertoires and performed immunohistochemistry (IHC) on CD8+ tumor-infiltrating lymphocytes (TILs) and PD-L1 expression in 28 patients who had paired samples. Through the comparison, the TCR clonality of BM was higher than lung tumor, indicating the distinct pattern of the stronger oligoclonal T cell expansion in BM; the primary tumor had a higher TMB than oligo-BM (13.9 vs 8.7 mutations, p = 0.019); CD8 + TILs of BM were significantly lower than lung tumor (10% vs 30%, p = 0.015), revealing the lower level of cytotoxic T cell infiltration; BM showed statistically equivalent level of PD-L1 compared with lung tumor (p = 0.722). We further investigated the potential biomarkers associated with overall survival (OS) after brain surgery. We found that higher TCR clonality was related to prolonged OS in EGFR-treated patients (HR 0.175, p < 0.001) but the worse outcomes in non-EGFR-treated (HR 2.623, p = 0.034). More CD8+ TILs were an independently positive indicator for OS, in EGFR-treated (HR 0.160, p = 0.001) and non-EGFR-treated patients (HR 0.308, p = 0.009). These findings provide a meaningful molecular and clinical understanding of lung carcinoma and brain oligo-metastasis.

Small-cell carcinoma of the esophagus (SCCE) is a rare and aggressive cancer. Although several consistent genomic changes were observed previously between SCCE and small-cell lung cancer (SCLC), detailed mutational landscapes revealing discrepancies in genetic underpinnings of tumorigenesis between these two cancers are scarce, and little attention has been paid to answer whether these genetic alterations were related to the prognosis. Herein by performing whole-exome sequencing of 48 SCCE and 64 SCLC tumor samples, respectively we have shown that the number of driver mutations in SCCE was significantly lower than in SCLC (p = 0.0042). In SCCE, 46% of recurrent driver mutations were clonal, which occurred at an early stage during tumorigenesis, while 16 driver mutations were found clonal in SCLC. NOTCH1/3, PIK3CA, and ATM were specifically clonal in SCCE, while TP53 was clonal in SCLC. The total number of clonal mutations differed between two cancers and presented lower in SCCE compared to SCLC (p = 0.0036). Moreover, overall survival (OS) was shorter in patients with higher numbers of clonal mutations for both cancers. In summary, SCCE showed distinct mutational background and clonal architecture compared with SCLC. Organ-specific clonal events revealed different molecular mechanisms underlying tumorigenesis, tumor development, patients' prognosis, and possible variations in therapeutic outcomes to candidate treatments.

BACKGROUND:Immune checkpoint inhibitors play a vital role in triple-negative breast cancer (TNBC) immunotherapy. A recent study showed that chemokine-like factor (CKLF)-like MARVEL transmembrane domain containing 6 (CMTM6) has a crucial role in programmed death-ligand 1 (PD-L1) stability. The aim of this study was to investigate the relationship between CMTM6 and PD-L1 in TNBC and the association with clinical characteristics.METHODS:A total of 143 patients, including 75 with human epidermal growth factor receptor 2 (HER2)-driven breast cancer and 68 with TNBC, were included in this study. In 83 paired primary breast cancers (PBCs) and metastatic breast cancers (MBC) comprising 45 HER2-driven breast cancers and 38 TNBC, CMTM6 and PD-L1 were detected based on immunohistochemistry (IHC) with FFPE tissues. Another 60 PBCs comprising 30 HER2-driven breast cancers and 30 TNBC in order to detect CMTM6 and PD-L1 mRNA expressions based on real-time polymerase chain reaction (RT-PCR) using frozen tissues. Furthermore, 153 patients comprising 30 TNBC and 123 HER2-driven breast cancer based on The Cancer Genome Atlas (TCGA) database were used to confirm the difference mRNA expression.RESULTS:The expression of CMTM6 in patients with TNBC was significantly higher than in those with HER2-driven PBC (IHC, P=0.036, mRNA, P=0.036, TCGA dataset, P=0.039). CMTM6 was correlated with PD-L1 based on IHC in triple-negative MBC (P=0.004); the same result was found based on mRNA data in triple- negative PBC (P=0.021). Moreover, a high expression of CMTM6 in TNBC was associated with poor progression-free survival (PFS) (P=0.030, 95% CI: 1.08-4.57, HR =2.22). After multiple Cox regression analysis, CMTM6 in TNBC emerged as an independent risk factor for PFS (P=0.027, 95% CI: 1.11-5.20, HR =2.40). The expression of PD-L1 was negatively correlated with lymph node metastasis (P=0.026) and was not associated with PFS.CONCLUSIONS:The expression of CMTM6 was higher in TNBC than in HER2-driven breast cancer. In TNBC, CMTM6 was correlated with PD-L1 expression, and potentially could be used as an independent risk factor for predicting PFS.

BACKGROUND:Esophageal carcinoma (EC) is the sixth most common cause of cancer-related mortality worldwide. Studying the associations of the tumor microenvironment (TME) with pathology and prognosis would illustrate the underlying mechanism of prognostic prediction and provide novel targets for immunotherapy in the treatment of EC.METHODS:Transcriptomic profiles of 159 EC patients were obtained from The Cancer Genome Atlas (TCGA) database. Stromal and immune scores were calculated using the ESTIMATE algorithm. Differentially expressed genes (DEGs) were identified by the optimal score cutoff. Functional enrichments were analyzed by DAVID, while prognostic genes were explored using the Kaplan-Meier method. Validation analysis was performed using immunohistochemistry in tissue microarrays containing samples from 145 EC patients. Multiplex immunofluorescence staining was performed to detect a panel of 6 immune markers, including T-cell immunoreceptor with Ig and ITIM domains (TIGIT), in 90 EC patients.RESULTS:Immune scores significantly increased with increasing age, while stromal scores were dramatically elevated with increasing tumor stage. Fifteen TME-related DEGs including allograft inflammatory factor 1 (AIF1) were identified as prognostic factors of EC. Furthermore, the validation cohort indicated that AIF1 was negatively associated with the prognosis of esophageal squamous cell carcinoma patients. Subsequent analyses suggested that AIF1 may affect immune infiltrates, including T cells and natural-killer cells. Moreover, a correlation between AIF1 and TIGIT was identified.CONCLUSIONS:These results indicate that the TME-related gene AIF1 is a promising predictor of prognosis and is related to immune infiltrates and TIGIT expression in EC. However, further mechanistic studies are needed.

目的 探讨表皮生长因子受体(EGFR)基因突变和间变性淋巴瘤激酶(ALK)重排在肺浸润性黏液腺癌(IMA)中的表达及与临床病理特征的关系.方法 选取2010年1月至2015年12月中国科学院大学附属肿瘤医院收治的IMA患者101例,应用突变扩增系统PCR法检测EGFR基因,VENTANA ALK免疫组化法检测ALK重排.结果 101例患者中共67例患者行EGFR和(或)ALK检测,其中EGFR/ALK均行检测32例,仅行EGFR检测20例,仅行ALK检测15例;34例患者未参与检测.EGFR突变阳性率为23.1％,在女性患者中表达率更高(P<0.05).ALK重排阳性率为27.7％,其表达情况与T分期有关,T分期越高越倾向于发生ALK重排(P<0.05).EGFR/ALK检测同时阳性占3.1％.结论 IMA患者的EGFR突变多见于女性,ALK重排与T分期有关.EGFR突变和ALK重排很少共存.

目的 探讨成纤维细胞生长因子受体4(FGFR4)在膀胱肌层浸润性尿路上皮癌(MIBUC)中的表达及意义.方法 采用免疫组化EnVision两步法以及Western blot方法检测50例MIBUC组织及其配对癌旁正常膀胱黏膜组织中FGFR4蛋白的表达,并分析其与MIBUC病理及预后的关系.结果(1)MIBUC组织中FGFT4蛋白表达高于癌旁正常膀胱黏膜组织(P<0.05);(2)FGFR4阳性表达与神经侵犯、淋巴结转移、肿瘤T分期有关(P<0.05),与患者的年龄、性别、肿瘤大小、脉管侵犯和组织学分级无明显关系(P>0.05);(3)FGFR4的表达上调可导致β-catenin异常表达.结论(1)FGFR4蛋白在MIBUC组织中呈高表达,其在MIBUC发生、发展中起重要作用;(2)β-catenin参与了上皮间质转化(EMT)的过程;(3)FGFR4蛋白阴性表达者预后好于阳性表达者,可作为临床上MIBUC预后的评价指标.

目的 探讨分化型甲状腺癌(DTC)神经内分泌分化(NED)与131I治疗疗效的关系.方法 收集46例1991年3月至2017年5月在中国科学院大学附属肿瘤医院行131I治疗伴随远处转移DTC患者的病变甲状腺组织石蜡标本,采用免疫组化法检测NED标志性蛋白嗜铬粒蛋白A(CgA)、神经细胞黏附分子1(CD56)、突触蛋白(Syn)、促甲状腺激素受体(TSHR)、钠碘同向转运体(NIS)的表达.采用χ2检验分析CgA、CD56、Syn表达与碘代谢关键基因TSHR、NIS表达的关系及CgA、CD56、Syn、TSHR、NIS的表达与临床特征、131I疗效的关系.结果 本组患者病灶摄碘率为60.8％(28/46),131I治疗有效率为36.9％(17/46).NIS、TSHR、CgA、CD56在DTC组织中的阳性表达率分别为28.2％(13/46)、15.2％(7/46)、8.6％(4/46)、15.2％(7/46),Syn无阳性表达.CgA与TSHR、NIS表达之间差异均无统计学意义(均P>0.05).CD56阳性患者中NIS阳性率为38.5％(5/13),高于CD56阴性的6.9％(2/29),差异有统计学意义(P<0.05),而CD56阳性与阴性患者TSHR表达率的差异无统计学意义(P>0.05).CgA、CD56表达水平在不同年龄、性别、单双侧、分期、远处转移及疗效的DTC患者之间差异均无统计学意义(均P>0.05);NIS表达水平在不同年龄DTC患者之间的差异有统计学意义(P<0.05),而在不同性别、单双侧、分期、远处转移、摄碘率及疗效的DTC患者之间差异均无统计学意义(均P>0.05);TSHR表达水平在不同性别的DTC患者之间差异有统计学意义(P<0.05),而在不同年龄、单双侧、分期、远处转移、摄碘率及疗效的DTC患者之间差异均无统计学意义(均P>0.05).结论 131I治疗可以使部分肺转移DTC患者获益,是远处转移DTC术后治疗的重要手段.部分DTC组织存在NED,CD56的表达与NIS表达之间差异有统计学意义,CD56可能对NIS的表达发挥作用,相关机制需进一步研究.

Small ubiquitin-like modifier 4 (SUMO4) is the latest member of the sumoylation family, which enhances the stability of protein, regulates the distribution and localization of the protein, and affects the transcription activity of the protein. However, the role of SUMO4 in non-small cell lung cancer (NSCLC) has not yet been reported. The present study first demonstrated that SUMO4 was upregulated in a number of tissues from patients with NSCLC. Immunohistochemistry was performed to demonstrate the expression level of SUMO4 in lung cancer tumor tissues. Following the transfection, The EMT status and signaling pathway activation regulated by SUMO4-siRNA was assessed by western blotting. The Transwell and wound healing assays were performed to investigate the regulatory effect of SUMO4-siRNA on cell migration and invasion. Cell Counting Kit-8 assay was performed to investigate whether SUMO4-siRNA affected the chemosensitivity of the NSCLC cells to cisplatin. Statistical analysis of immunohistochemical results from the tissues showed that the overexpression of SUMO4 was significantly associated with sex, tumor type, history of smoking, T stage and poor prognosis. It was also identified that SUMO4 small interfering RNA attenuated invasion and migration in NSCLC cell lines, as well chemosensitivity to cisplatin via the inhibition of the JAK2/STAT3 pathway. In conclusion, SUMO4 may play an important role in the poor prognosis of patients with NSCLC. The present study indicates that SUMO4 may be a potential therapeutic target for NSCLC.

Extracellular vesicles (EVs) are excellent potential vectors for the delivery of therapeutic drugs. However, issues with biological safety and disease targeting substantially limit their clinical application. EVs from red blood cells (RBC-EVs) are potential drug delivery vehicles because of their unique biological safety. Here, we demonstrated that EVs, including RBC-EVs, show natural liver accumulation. Mechanistically, the liver environment induces macrophages to phagocytize RBC-EVs in a C1q-dependent manner. RBC-EVs loaded with antisense oligonucleotides of microRNA-155 showed macrophage-dependent protective effects against acute liver failure (ALF) in a mouse model. These RBC-EVs were also effective in treatment of ALF. Furthermore, compared to routine doses of doxorubicin and sorafenib (SRF), RBC-EVs loaded with doxorubicin or SRF showed enhanced therapeutic effects on a murine model of orthotopic liver cancer through a mechanism dependent on macrophages. Importantly, drug-loaded RBC-EVs showed no systemic toxicity at therapeutically effective doses, whereas routine doses of doxorubicin and SRF showed obvious toxicity. Thus, drug-loaded RBC-EVs hold high potential for clinical applications in the treatment of liver disease therapy.

Background: Graded prognostic assessment for lung cancer using molecular markers (Lung-molGPA) for brain metastases is a powerful prognostic tool. However, it has not been validated for non-small-cell lung cancer (NSCLC) patients with synchronous or meta-chronous brain metastases. Methods: A total of 1184 NSCLC patients with synchronous or metachronous brain metastases were reviewed in this study. Comparative clinicopathological variables and survival analysis for these two groups (synchronous vs metachronous), as well as complimentary analysis of prognostic factors for the entire patient cohort, were performed. Afterward, patients were stratified using Lung-molGPA to evaluate the accuracy of the survival estimates. Results: A total of 763 patients (64.4%) had synchronous metastases while 35.6% (421 patients) had metachronous metastasis. Patients with synchronous metastases were more likely to have a smoking history, limited metastatic lesions, and absence of cerebral symptoms (P<0.05). Patients with metachronous metastatic NSCLC had an overall survival (OS) period of 16.5 (95% CI 14.5-18.6) months and were longer compared to patients with synchronous metastases (16.5 vs 13.5 [12.5-14.6] months, P=0.004). In Cox regression multivariable analysis, age (HR=1.25, P=0.008), Karnofsky performance status (HR=1.30, P=0.005), extracranial metastases (HR=1.57, P<0.001), number of brain metastases (HR=1.22, P=0.043), gene mutation (HR=1.40 [wild type vs mutation], P=0.050; HR=1.42 [unknown vs mutation], P=0.007), and treatment (including TKI, chemotherapy, and local brain treatment, P<0.05) were independent prognostic predictors of OS. Additionally, metachronous metastatic patients were at lower risk for disease-related death compared to synchronous metastatic patients (HR=0.69, P<0.001). Importantly, median OS stratified by Lung-molGPA of 0-1, 1.5-2, 2.5-3 and 3.5-4 scores were 11.0, 14.0, 24.9, and 26.3 months for synchronous brain metastases, and 13.1, 17.0, 37.2, and 66.5 months for metachronous metastases, respectively (P<0.001). Conclusion: Lung-molGPA could estimate the prognosis of NSCLC patients with synchronous or metachronous brain metastases. Hence, patients should be carefully stratified for consideration of aggressive therapy.

Adequate lymphadenectomy is critical for accurate nodal staging and planning adjuvant therapy in colon cancer. However, the optimal lymph node (LN) yield for stage II right-sided colon cancer (RSCC) is still unclear. This population-based study aimed to determine the optimal LN yield associated with survival and LN positivity in patients with stage II RSCC. All patients with stage II–III RSCC were identified from the Surveillance, Epidemiology, and End Results database over a 10-year interval (2006–2015). The optimal threshold for LN yield was explored using an outcome-oriented approach based on survival and LN positivity. The median number of LNs examined for all 17,385 patients with stage II RSCC was 17 (IQR 12–23). Nineteen LNs were determined as the optimal cut-off point to maximize survival benefit from lymphadenectomy. Increased LN yield was associated with a gradual increase in the risk of node positivity, with no change after 19 nodes. Compared with patients with 19 or more LNs examined, the group with fewer LNs had a significantly poor cancer-specific survival (< 12 nodes: hazard ratio (HR) 2.26, P < 0.001; 12–18 nodes: HR 1.58, P < 0.001) and overall survival (< 12 nodes: HR 1.80, P < 0.001; 12–18 nodes: HR 1.31, P < 0.001). Similar survival results were found in the validation cohort. Patients with older age, small tumor size, and appendix and transverse colon cancer were more likely to receive inadequate LN harvest. A minimum of 19 LNs is needed to be examined for optimal survival and adequate node staging in lymph node-negative RSCC.