In recent years, there have been breakthroughs in the preclinical research of respiratory diseases, such as organoids and organ tissue chip models, but they still cannot provide insight into human respiratory diseases well. Human lung slices model provides a promising in vitro model for the study of respiratory diseases because of its preservation of lung structure and major cell types. Human lung slices were manually prepared from small pieces of lung tissues obtained from lung cancer patients subjected to lung surgery. To evaluate the suitability of this model for lung fibrosis research, lung slices were treated with CdCl2 (30 μM), TGF-β1 (1 ng/ml) or CdCl2 plus TGF-β1 for 3 days followed by toxicity assessment, gene expression analysis and histopathological observations. CdCl2 treatment resulted in a concentration-dependent toxicity profile evidenced by MTT assay as well as histopathological observations. In comparison with the untreated group, CdCl2 and TGF-β1 significantly induces MMP2 and MMP9 gene expression but not MMP1. Interestingly, CdCl2 plus TGF-β1 significantly induces the expression of MMP1 but not MMP2, MMP7 or MMP9. Microscopic observations reveal the pathogenesis of interstitial lung fibrosis in the lung slices of all groups; however, CdCl2 plus TGF-β1 treatment leads to a greater alveolar septa thickness and the formation of fibroblast foci-like pathological features. The lung slice model is in short of blood supply and the inflammatory/immune-responses are considered minimal. The results are in favor of the hypothesis that idiopathic pulmonary fibrosis (IPF) is mediated by tissue damage and abnormal repair. Induction of MMP1 gene expression and fibroblast foci-like pathogenesis suggest that this model might represent an early stage of IPF.

Previous studies have demonstrated the tumor-suppressive function of microRNA-22-3p (miR-22-3p) in several cancers, whereas the significance of miR-22-3p in non-small cell lung cancer (NSCLC) remains unclear. In this study, we explored the biological function and molecular mechanism of miR-22-3p in NSCLC cells. First, we assessed the expression of miR-22-3p in NSCLC tissues and cells based on RT-qPCR and TCGA database. Compared with normal lung tissues and cells, miR-22-3p expression was dramatically decreased in lung cancer tissues and cells. miR-22-3p expression was also correlated with lymph node metastasis and tumor size, but not TNM stages. We further explored the in vitro function of miR-22-3p on the migration and epithelial-mesenchymal transition (EMT) of NSCLC cells. The results showed that overexpression of miR-22-3p suppressed the migration and EMT of NSCLC cells, whereas silencing miR-22-3p showed the opposite effect. Luciferase assay demonstrated that RAS-related C3 botulinum toxin substrate 1 (RAC1) was the target gene for miR-22-3p. Mechanistically, we demonstrated that miR-22-3p suppressed the cell migration and EMT via downregulation of RAC1 because the inhibitory effect of miR-22-3p on cell migration and EMT of NSCLC cells was reversed by RAC1 overexpression. Based on these novel data, the miR-22-3p/RAC1 axis may be an alternative target in the therapeutic intervention of NSCLC.

目的:分析比较表皮生长因子受体酪氨酸激酶抑制剂(EGFR-TKIs)联合化疗与EGFR-TKIs单药一线治疗晚期非小细胞肺癌(NSCLC)的疗效和安全性.方法:回顾性分析2010年09月至2015年12月晚期EGFR突变阳性NSCLC患者100例,根据治疗方案分为联合组(50例)和单药组(50例),联合组采用EGFR-TKIs联合化疗治疗,单药组采用EGFR-TKIs单药治疗.比较两组患者的治疗效果、预后情况以及毒副作用.结果:联合组总有效率为66％(33/50),单药组为48％(24/50),组间比较差异无统计学意义(P=0.106);联合组的疾病控制率为88％(44/50),单药组的疾病控制率为68％(34/50),组间比较差异有统计学意义(P=0.028).联合组和单药组的中位PFS分别为18.5个月和13.5个月,差异有统计学意义(P=0.013),两组的OS分别为36个月和28.5个月,差异无统计学意义(P=0.071).单药组和联合组的3-4级治疗相关不良反应发生率分别为6％和10％,差异无统计学意义(P=0.463).结论:与EGFR-TKIs单药相比,EGFR-TKIs联合化疗一线治疗晚期EGFR基因突变NSCLC患者,可提高PFS,且不良反应可耐受.

Background Histone deacetylases (HDACs) play crucial roles in cancers, but the role and mechanism of HDAC7 in NSCLC have not been fully understood. Methods A total of 319 patients with non-small cell lung cancer (NSCLC) who underwent surgery were enrolled in this study. Immunohistochemistry and Kaplan–Meier survival analysis were performed to investigate the relationship between HDAC7, fibroblast growth factor 18 (FGF18) expression, and clinicopathologic characteristics. Cell functional experiments were implemented both in vivo and in vitro to investigate the effects on NSCLC cell proliferation and metastasis. Recombinant lentivirus–meditated in vivo gene overexpression or knockdown, real-time polymerase chain reaction (PCR), western blotting, and coimmunoprecipitation assays were applied to clarify the underlying molecular mechanism of HDAC7 in promoting NSCLC progression. Results The elevated expression of HDAC7 or FGF18 was positively correlated with poor prognosis, tumor–node–metastasis (TNM) stage, and tumor differentiation of NSCLC patients. NSCLC patients with co-expressed HDAC7 and FGF18 suffered the worst prognosis. HDAC7 overexpression promoted NSCLC proliferation and metastasis by upregulating FGF18. Conversely, overexpression of FGF18 reversed the attenuated ability in tumor growth and metastasis mediated by downregulating HDAC7. In terms of mechanism, our results suggested that the interaction of HDAC7 with β-catenin caused decreased β-catenin acetylation level at Lys49 and decreased phosphorylation level at Ser45. As a consequence, the HDAC7-mediated posttranslational modification of β-catenin facilitated nuclear transfer and activated FGF18 expression via binding to TCF4. Furthermore, deubiquitinase USP10 interacted with and stabilized HDAC7. The suppression of USP10 significantly accelerated the degradation of HDAC7 and weakened NSCLC growth and migration. Conclusions Our findings reveal that HDAC7 promotes NSCLC progression through being stabilized by USP10 and activating the β-catenin-FGF18 pathway. Targeting this novel pathway may be a promising strategy for further developments in NSCLC therapy.

Aims Serine/threonine/tyrosine kinase 1 (STYK1) has been previously shown to have oncogenic properties, and emerging evidence suggests that STYK1 expression correlates with epithelial-mesenchymal transition (EMT). However, the mechanism of STYK1 involvement in oncogenesis remains unknown. The present study aimed to elucidate how STYK1 expression level relates to the metastasis, migration, invasion, and EMT in non-small cell lung cancer (NSCLC) and to determine the molecular mechanism of STYK1 effects. Methods Serine/threonine/tyrosine kinase 1 (STYK1) expression level and its relationship with the prognosis of NSCLC were determined using the ONCOMINE database and clinical cases. Non-small cell lung cancer cell lines with the overexpression or knockdown of STYK1 were established to determine whether STYK1 promotes cell migration, invasion, and EMT in vitro and in vivo. In addition, a constitutively active FoxO1 mutant (FoxO1AAA) was used to examine the role of FoxO1 in the STYK1-mediated upregulation of metastasis and EMT in NSCLC. Results Serine/threonine/tyrosine kinase 1 (STYK1) was upregulated in NSCLC tissues and cell lines, and its overexpression correlated with poor prognosis in patients with NSCLC after surgery. Enhanced expression of STYK1 potentiated the migration, invasion, and EMT in SW900 cells, thereby promoting metastasis, whereas knockdown of STYK1 inhibited these cellular phenomena in Calu-1 cells. Furthermore, STYK1 expression was positively related to the level of phosphorylated-FoxO1, whereas the constitutively active FoxO1 mutant protected against the positive effect of STYK1 overexpression on cell migration, invasion, and EMT. Conclusion Serine/threonine/tyrosine kinase 1 (STYK1) was upregulated in NSCLC and correlated with poor clinical outcomes. In addition, STYK1 suppressed FoxO1 functions, thereby promoting metastasis and EMT in NSCLC.

目的 探讨来自中国西北地区汉族人群的非小细胞肺癌(NSCLC)患者表皮生长因子受体(EGFR)、间变性淋巴瘤激酶(ALK)和C?ros原癌基因1?受体酪氨酸激酶(ROS1)基因突变特点及与临床病理特征的相关性.方法 收集2014年12月至2018年10月空军军医大学第二附属医院胸腔外科就诊的4580例NSCLC患者的肿瘤组织标本及临床资料,采用扩增阻滞突变系统(ARMS)检测EGFR基因的突变状态及ALK和ROS1融合基因表达情况,并分析其与临床病理特征的关系.结果 4580例NSCLC中EGFR基因突变率为38.30％(1754/4580),19?del和L858R突变为其主要突变类型,EGFR外显子双重突变率为3.14％(55/1754).EGFR基因在女性、腺癌、非吸烟、无淋巴结转移及早期患者中的突变率较高(P<0.001).ALK融合基因突变率为5.96％(273/4580),ROS1融合基因突变率为1.38％(63/4580),在女性、年龄≤60岁、腺癌、非吸烟、有淋巴结转移及晚期的患者中ALK与ROS1融合基因突变的发生率较高(P<0.001).EGFR和ALK基因突变共存29例,EGFR和ROS1基因突变共存3例.结论 NSCLC患者中EGFR基因突变率明显高于ALK及ROS1融合基因突变率.EGFR、ALK和ROS1基因突变均易发生于女性、腺癌及非吸烟患者中,但EGFR基因突变更易发生于早期患者中,而ALK和ROS1融合基因突变更多发生于晚期患者中.EGFR基因突变和ALK或ROS1融合基因突变均可共存.

Background: For low-risk areas, it was vital to the management of coronavirus disease 2019 (COVID-19) patients who had history of exposure in epicenter, however, most of them could not provide the clear exposure time. So this study was designed to predict the progression of COVID-19 by using the definite departure time as original observation point. Methods: In this retrospective study,all cases were selected from the northwest regions with the lowest population density in China to reduce the possibility of secondary exposure. As our study endpoints,we defined the incubation period as from the date of departure of Wuhan City to the date of symptom onset or the date of confirmed time(just for asymptomatic patients), the confirmed time was defined as the interval of symptom onset to the positive results (samples from the respiratory tract). Both of them were estimated by fitting a Weibull distribution on the departure date and symptom onset. The difference among the variables were analyzed. Findings: 139 patients (98 were residents and 41 patients were travellers) were finally enrolled, about 10.1% patients (14 patients) were absent of symptom during their disease course. We estimated the median incubation period was 4.0 days (interquartile intervals, 2.0 to 8.0), the 95th percentile of the distribution was 15 days. Moreover, about 75.2% symptomatic patients had onset of symptoms within a week (94/125), however, 5.6% patients (7 patients) experienced symptoms two weeks after leaving,for one patient developed after 23.0 days. Furthermore,we explored the issue about the optimal detection time after the symptom onset,the estimation median interval from symptom onset to final diagnosis was 4.0 days (interquartile intervals,2.0 to 6.0),the 95th percentile of the distribution was 12.0 days. Further analysis showed that old patients were detected positive result later than young (5.0 days vs 4.0 days,P=0.028). Finally,the median hospitalization time was 16.0 days, ranged from 3.0 days to 45.0 days. Univariate analysis showed that age (P=0.021) and severity status (P=0.001) were correlated significantly with hospitalization time. Interpretation: This retrospective study showed the median incubation period of COVID-19 was 4.0 days after they leaved outbreak area,the median interval from symptom onset to final diagnosis was also 4.0 days. Approximately 5.6% patients experienced symptoms two weeks after leaving Wuhan City, so the quarantine period of two weeks need to be prolonged.It was worth noting that 10.1% asymptomatic patients were detected positive results on the samples from the respiratory tract, so they should not be ignored. Funding Statement: This research was supported by grants from the Wu Jieping Medical Foundation (320.6750.17527) and Provincial Key R&D Program of Shaanxi Province (2017ZDCXL-SF-01-04-01).Declaration of Interests: We declared that we have no conflicts of interest,the opinions expressed by authors contributing to this journal do not necessarily reflect the opinions of the Centers for Disease Control and Prevention or the institutions with which the authors are affiliated.

BACKGROUND:Some non-small-cell lung cancer (NSCLC) patients are unexpectedly diagnosed with stage IIIA-N2 disease at the time of thoracoscopy or thoracotomy. Because of the limited statistical evidence of induction chemotherapy for these patients, it is necessary to develop more profound treatment strategies.METHODS:The demographic and clinical characteristics of patients with stage IIIA-N2 NSCLC harboring epidermal growth factor receptor (EGFR) mutations after radical resection were retrospectively reviewed. The patients were divided into 3 groups based on treatment: EGFR tyrosine kinase inhibitors (EGFR-TKIs, erlotinib or gefitinib), adjuvant chemotherapy (docetaxel plus cisplatin), and combination treatment (chemotherapy plus EGFR-TKIs). The effect of adjuvant therapy on survival rate was assessed using univariate and Cox regression analyses.RESULTS:Patients receiving EGFR-TKIs alone showed significantly improved disease-free survival (DFS; p = 0.025) when compared to those receiving chemotherapy alone. Compared to chemotherapy alone, the combination of chemotherapy and EGFR-TKIs resulted did not significantly improve DFS (p < 0.001) and overall survival (OS p < 0.001). The combination of EGFR-TKIs with chemotherapy as adjuvant therapy led to improvements in both DFS (p = 0.116) and OS (p = 0.039) compared to patients receiving a EGFR-TKI monotherapy. Toxicities were mild in the 3 treatment groups.CONCLUSIONS:Our study demonstrated that adjuvant EGFR-TKI treatment significantly increased the DFS of patients with stage IIIA-N2 NSCLC when compared with cisplatin-based chemotherapy. The use of EGFR-TKIs and chemotherapy is recommended in the setting of combined-modality therapy.

Background Grade prognostic assessment (GPA) is widely used to evaluate the prognosis of non-small cell lung cancer (NSCLC) patients with brain metastases (BMs). This study aimed to investigate whether lymph node status (LNS) could be included as one of the GPA variables for NSCLC with BMs. Methods Overall, 586 patients with NSCLC and BMs were retrospectively analyzed. Overall survival stratified by LNS was analyzed using the Kaplan-Meier method. Multivariate analysis was also performed to identify independent prognostic factors using the Cox proportional hazards progression model. In the updated GPA index, prognostic factors and criteria of GPA score were weighted by effect magnitude relative risk (RR) and statistical significance. Results In NSCLC patients with BMs, those with lymph node involvement had worse overall survival (mOS, 13.4 months vs. 25.9 months, P <0.001) than those without lymph node involvement. Multivariate analysis showed that LNS might be an independent prognostic factor (RR: 1.702, CI: 1.340-2.162, P <0.001). Finally, five prognostic factors including LNS, the age of the patient, Karnofsky performance status (KPS), the number of BMs, and extracranial metastases were enrolled in our novel GPA index. With the updated GPA index involving the N stage, survival analysis was also performed. Prognostic results were significantly different among these four subgroups (Class A vs. Class B, P=0.047; Class B vs. Class C, P<0.001; Class C vs. Class D, P=0.007). Conclusions These results indicate that LNS might be an indispensable prognostic factor in NSCLC with BM. The novel GPA model involving the N stage could provide more reliable evidence to estimate the survival of NSCLC patients with BMs.

Huntingtin interacting protein 1 (HIP1) is over-expressed in several human malignancies. However, the biological function of HIP1 in esophageal squamous cell carcinoma (ESCC), and its effect on the prognosis of patients remain unclear. The present study aimed to investigate HIP1 expression in ESCC via immunohistochemistry, reverse transcription-quantitative PCR and western blot analyses. The association between HIP1 expression and the clinicopathological characteristics of 173 patients with ESCC was statistically analyzed. The effect of HIP1 expression on patient prognosis was assessed via Kaplan-Meier and Cox regression analyses. Lentivirus-delivered RNA interfering technique was used to overexpress and downregulate HIP1 expression in ESCC cell lines. The results demonstrated that HIP1 expression was significantly higher in ESCC tissues compared with adjacent normal tissues, and HIP1 expression was associated with histological differentiation, tumor-node-metastasis stage and lymph node metastasis. Furthermore, the overall survival time of patients with high HIP1 expression was significantly shorter than those with low HIP1 expression. Cellular mobility demonstrated that overexpressing HIP1 increased ESCC proliferation, migration and invasion, whereas silencing HIP1 decreased ESCC proliferation, migration and invasion. Furthermore, overexpressing HIP1 induced ESCC cells to enter the S and G2 phases from the G1 phase, whereas HIP1 knockdown arrested the cell cycle in the G1 phase. Taken together, the results of the present study suggest that HIP1 is associated with proliferation and metastatic behaviors in ESCC, and thus may be used as a potential prog-nostic indicator for patients with ESCC.

[目的]探讨亨廷顿蛋白相互作用蛋白1 (huntingtin interacting protein 1,HIP1)促进食管鳞癌转移的作用及机制.[方法]采用qRT-PCR及Western blot检测Akt抑制剂处理HIP1高或低表达食管鳞癌细胞系中HIP1、GSK3β及EMT标志性分子E-cadherin和Vimentin的表达.采用IHC检测食管鳞癌组织样本中目的分子的表达,并分析其表达相关性.[结果] qRT-PCR和Western blot结果显示:与对照组、shRNA-对照组相比,shRNA-HIP1组细胞中Akt、GSK3β及Vimentin基因和蛋白表达显著降低(P＜0.001),E-cadherin基因和蛋白表达显著升高(P＜0.001);与对照组、OE-对照组相比,OE-HIP1组细胞中Akt、GSK3β及Vimentin基因和蛋白表达显著升高(P＜0.001),E-cadherin基因和蛋白表达显著降低(P＜0.001).IHC及相关性分析结果显示,HIP1与GSK3β、Vimentin表达呈正相关(r=0.336,P＜0.001;r=0.561,P＜0.001),与E-Cadherin表达呈负相关(r=-0.169,P=0.027);GSK3β与Vimentin表达呈正相关(r=0.317,P＜0.001),与E-Cadherin表达呈负相关(r=-0.171,P=0.025).Akt抑制剂处理HIP1高或低表达食管鳞癌细胞系结果显示,Akt抑制剂处理后E-cadherin表达升高,GSK3β及Vimentin表达降低.[结论]HIP1可能通过激活Akt/GSK3β信号通路促进食管鳞癌EMT的发生.

The evaluation of EGFR mutation genes in circulating tumor DNA (ctDNA) in blood sample is key for patients with lung cancer. Surface-enhanced Raman scattering (SERS) has potential for trace detection of DNA or RNA. The detection rate offered by current methods can not meet clinical demand. By combining asymmetric polymerase chain reaction (PCR) and SERS, a highly-selective detection for EGFR mutation genes in lung cancer was developed. Sea-urchin like Au nanoclusters (AuNCs) were synthesized via Ag seed-mediated growth. AuNCs with a diameter of 120 nm were covered with 79 nanopricks (20 nm). Then, EGFR mutation specific molecular beacons (MBs) labeled with Cy3 were coated on the surface of AuNCs. The loading amount of MBs was calculated as 5720 ± 740 on one AuNCs. These AuNCs probes had good efficiency (equilibrium time: 20 minutes) with high sensitivity (detection limit: 5.8 nM), high specificity (capable of single-base mismatch recognition) and good stability against nucleases. Following this, asymmetric PCR was performed to obtain large numbers of single-stranded DNA (ssDNA, E746-A750del). The ssDNA was incubated with the AuNCs probes and tested quantitatively based on the SERS signals of the AuNCs probes. This combined asymmetric PCR-SERS method had a very high detection threshold (4.24 fM). The asymmetric PCR-SERS method was shown to have an overall sensitivity of 75% and specificity of 100% in a further 15 clinical blood samples. This method is proved to be promising for non-invasive and sensitive detection of EGFR mutations in ctDNA.

Background: Few studies have reported the transmission characteristics of coronavirus disease (COVID-19) in low-density populations. This study has therefore analyzed the epidemiological characteristics and clinical outcomes of COVID-19 patients in Northwestern China, an area with low population density. Methods: From January 21 to March 11, 2020, data from patients diagnosed with novel coronavirus pneumonia (NCP) in areas of Northwestern China with lower population densities were retrospectively analyzed. Certain variables were categorized as numbers and percentages, with the ratio between resident patients (no history of going out during the epidemic) and imported patients representing the contagiousness of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) responsible for COVID-19. Hospitalization time was also calculated. Results: A total of 617 COVID-19 patients were reported in Northwestern China, and the morbidity and mortality rates of 0.000005 and 0.011, respectively. Further analysis showed that the morbidity was inversely proportional to population density and distance from Wuhan City. This study enrolled 473 confirmed cases; among these patients, there were 248 residents and 225 imported cases with a ratio of 1:1. The youngest and oldest patients were 1 and 94 years of age, respectively, with a median age of 42 years. Fifteen (3.2%) patients were children or infants. Two patients were pregnant, and one patient gave birth to a healthy baby with negative results during her disease course. About 17.3% of patients (82 cases) were healthy carriers without any symptoms during their disease course. One male patient (0.2%) had recurrence of a positive test result 4 days after discharge. The median hospitalization time was 16.0 days, ranging from 2.0 to 43.0 days. Further analysis showed that age (P = 0.03) and severity status (P < 0.001) were significantly correlated with hospitalization time. Conclusions: The morbidity and mortality rates of COVID-19 patients in the regions with a low population density were lower than those of the national average in China. All populations were susceptible to infection by SARS-CoV-2. Asymptomatic patients with positive results should be taken seriously, and the hospitalization time of patients is associated with their age and severity status.

Background CHCHD2 was identified a novel cell migration-promoting gene, which could promote cell migration and altered cell adhesion when ectopically overexpressed in NIH3T3 fibroblasts, and it was identified as a protein necessary for OxPhos function as well. However, the clinic relevance of CHCHD2 expression in NSCLC remains unclear. Here we assumed that CHCHD2 expression would accompanies the expression of HIF-1α to response hypoxia in the occurrence of NSCLC. Methods In order to verify this hypothesis, correlations among the expression levels of CHCHD2 and HIF-1α were detected and analyzed in 209 pair cases of NSCLC. The expression and location of these molecules were assessed using Immunohistochemistry, immunohistofluorescence, qRT-PCR and western blotting. The differences and correlations of the expression of these two molecules with clinical pathological characteristics in NSCLC were statistically analyzed using Wilcoxon (W) text, Mann-Whitney U, Kruskal-Wallis H and cross-table tests. Kaplan-Meier survival analysis and Cox proportional hazards models were used to estimate the effect of the expression of CHCHD2 and HIF-1α on the patients’ survival. Results Data showed that CHCHD2 and HIF-1α expression were higher in NSCLC than in normal tissues (all P = 0.000). CHCHD2 expression was significantly related with smoking, tumor size, differentiation degree, TNM Stage, lymph metastasis (all P< 0.05). The HIF-1α expression was significantly associated with smoking, tumor category, differentiation degree, TNM Stage, Lymph metastasis (all P< 0.05). There was a marked correlation of CHCHD2 and HIF-1α expression with histological type, differentiation and lymph metastasis of NSCLC (all P< 0.05, r s > 0.3). Immunohistofluorescence showed that there were co-localization phenomenon in cytoplasm and nucleus between CHCHD2 and HIF-1α expression. NSCLC patients with higher CHCHD2 and HIF-1α expression had a significantly worse prognosis than those with lower CHCHD2 and HIF-1α expression (all P = 0.0001; log-rank test). The multivariate analysis indicated that CHCHD2 expression was an independent prognostic factor in NSCLC (hazard ratio [HR], 0.492, P = 0.001). Conclusion Our results indicate that over-expression of CHCHD2 would promote the expression of HIF-1α to adapt the hypoxia microenviroment in NSCLC and CHCHD2 could serves as a prognostic biomarker in NSCLC.

Non-small-cell lung cancer (NSCLC) represents most of lung cancers, is often diagnosed at an advanced metastatic stage. Therefore, exploring the mechanisms underlying metastasis is key to understanding the development of NSCLC. The expression of B cell receptor-associated protein 31 (BCAP31), calreticulin, glucose-regulated protein 78, and glucose-regulated protein 94 were analyzed using immunohistochemical staining of 360 NSCLC patients. It resulted that the high-level expression of the four proteins, but particularly BCAP31, predicted inferior overall survival. What's more, BCAP31 was closely associated with histological grade and p53 status, which was verified by seven cohorts of NSCLC transcript microarray datasets. Then, three NSCLC cell lines were transfected to observe behavior changes BCAP31 caused, we found the fluctuation of BCAP31 significantly influenced the migration, invasion of NSCLC cells. To identify the pathway utilized by BCAP31, Gene Set Enrichment Analysis was firstly performed, showing Akt/m-TOR/p70S6K pathway was the significant one, which was verified by immunofluorescence, kinase phosphorylation and cellular behavioral observations. Finally, the data of label-free mass spectroscopy implied that BCAP31 plays a role in a fundamental biological process. This study provides the first demonstration of BCAP31 as a novel prognostic factor related to metastasis and suggests a new therapeutic strategy for NSCLC.

Background: Public health interventions presented effectiveness in controlling coronavirus infected disease 2019(COVID-19),most studies focused on the outbreak area or the high density population regions,but the related study in low density population areas was limited. So we designed this study to research the effect of public health intervention of COVID-19 in Northwestern China with low density population.Methods: From January 21st to March 11st 2020, all patients diagnosed with novel coronavirus pneumonia(NCP) in Northwestern China with a lower population density were retrospectively reviewed.Categorical variables were summarized as numbers and percentages,the ration between the local cases(no history of leaving Northwestern China) and imported cases can represent the contagious of COVID-19 virus. Hospitalization time was assessed as well.Findings: A total of 617 COVID-19 patients were reported in Northwestern China,the morbidity and mortality were 0.000005 and 0.011, respectively,further analysis showed that the morbidity was inversely proportional to population density and distance from Wuhan City. In this study, 473 confirmed cases were enrolled to be analyzed, for these patients, 248 patients were local cases and 225 patients were imported cases, the ration between them was 1.1. When the public health interventions performed, we found that four days after Wuhan City shutdown, the number of newly increased imported patients reached to peak with 22 cases, about 76.9% of imported cases (173/225) were diagnosed within 10 days of the closure of Wuhan City. To more powerfully prevent and control the COVID-19 spread, strictly countermeasures designed and approved by local authorities were performed. Four days after implement of controlled measures, the number of imported and local patients had significantly decreased, no newly confirmed patient occurred after 19 days hard work. Moreover, asymptomatic suspicious cases underwent nucleic acid testing, about 17.3% patients(82 cases) were detected positive results, they were healthy carriers, all of them were detected during the period of quarantine or medical observation. We also found that infants or children (less than 12 years) did not belong to a disease-exempt population,15 infants or children (3.25)were diagnoses with COVID-19. Finally,the median hospitalization time was 16.0 days,rang from 2.0 days to 43.0 days, further analysis showed that the following variables were correlated significantly with hospitalization time: age(P=0.03) and severity status(P<0.001).Interpretation: Both morbidity and mortality rate of COVID-19 patients in Northwestern China were lower than that of the national average of China,it might attributed to low population density and far away from epicenter.The public health interventions,as well as the management of asymptomatic patients and children,showing significance in controlling the outbreak.The hospitalization time of patients were associated with their age and severity status.Funding Statement: We declared that we have no conflicts of interest,the opinions expressed by authors contributing to this journal do not necessarily reflect the opinions of the Centers for Disease Control and Prevention or the institutions with which the authors are affiliated.Declaration of Interests: This research was supported by grants from the Wu Jieping Medical Foundation (320.6750.17527) and Provincial Key R&D Program of Shaanxi Province(2017ZDCXL-SF-01-04-01).

目的 本研究旨在探讨非小细胞肺癌(non-small cell lung cancer,NSCLC)中p44/WDR77表达状况及其与临床病理分化、肿瘤TNM分期及患者预后之间的关系.方法 采用免疫组化EnVinsion法检测p44/WDR77在192例患者NSCLC组织和癌旁正常组织中的表达情况,分析其表达与NSCLC病理分化、肿瘤TNM分期及患者预后之间的关系.结果 在192例NSCLC患者肿瘤组织中,p44/WDR77的阳性表达率为81.3％(156/192),在与之对应的癌旁正常组织中的阳性表达率为5.2％(10/192),二者比较差异显著(P<0.001).p44/WDR77在肺癌组织中高表达主要定位于胞核(鳞癌)和胞质(腺癌)中.鳞癌和腺癌组织中p44/WDR77的阳性表达率比较差异无统计学(P=0.067);p44/WDR77在高、中、低三种不同病理分化的NSCLC组织中阳性表达率分别为58.3％(7/12)、79.9％(115/144)、94.4％(34/36),差异有统计学意义(P=0.003);在临床肿瘤TNM不同分期的NSCLC组织中,p44/WDR77的阳性表达率分别为Ⅰ期77.1％(27/35)、Ⅱ期73.1％(49/67)、Ⅲ期85.7％(60/70)、Ⅳ期100％(20/20),表达差异显著(P<0.001);p44/WDR77的表达差异与患者的性别,解剖学肿瘤分类(中心型、周围型)均无统计学意义(P=0.861、P=0.462),而与NSCLC患者年龄、淋巴结转移与否、病灶范围显著相关(P<0.001、P=0.036、P=0.041).在随访的192例NSCLC病例中,p44/WDR77表达阳性和阴性的中位生存期(median survival time,MST)分别为21.3和37.5个月,差异具有统计学意义(P=0.049).结论 检测p44/WDR77的表达水平可能在NSCLC的诊断、恶性程度的预判、病灶转移及生存预后方面的预测具有重要的意义,有望成为诊断NSCLC和患者预后的潜在肿瘤标志物.

Objective To investigate the relationship between c-ros oncogene 1 (ROS1) fusion gene mutation and epidermal growth factor receptor (EGFR) mutation and clinicopathological characteristics in non-small cell lung cancer (NSCLC). Methods Real-time fluorescence quantitative PCR (QPCR) was performed to examine gene rearrangement of ROS1 fusion gene in 3487 NSCLC patients of Northwest China from December 2014 to December 2017. EGFR mutation was detected by ARMS method for patients with ROS1 fusion gene mutation. The clinicopathological features of patients with double mutations were analyzed. Results Among the 3487 patients, 54 patients (1. 5％) occurred ROS1 fusion gene mutation. ROS1 fusion gene mutation was associated with age, gender, smoking history, pathological types and clinical stage (P＜0. 05). Three patients were identified with EGFR mutation from 54 patients who harboring ROS1 fusion genes mutation, including 2 cases of EGFR19 exon deletion mutation (19-del), and 1 case of EGFR L858 R mutations. The 3 double mutative cases were of ROS1 variant 2 (R2). Conclusion ROS1 fusion gene mutative rate of NSCLC patients in Northwest China is 1. 5％. ROS1 fusion gene and EGFR mutations can coexist in NSCLC.

Serine Threonine Tyrosine Kinase 1 (STYK1) presents oncogenic properties in many studies, and emerging evidence suggests that ferroptosis serve as a novel tumor suppressor. However, the interplay between STYK1 and ferroptosis in NSCLC remains unclear. Our aim is to illustrate the expression of ferroptotic regulator Glutathione peroxidase 4 (GPX4) in NSCLC and the relationship between STYK1 and ferroptosis. Herein, results based on ONCOMINE database, clinical specimens, and cellular manipulation revealed GPX4 was upregulated in NSCLC tissues and cell lines, and high GPX4 expression predicted worse prognosis. High STYK1 expression predicted worse OS and was related to high GPX4 in NSCLC tissues; overexpression of STYK1 in lung cancer cell line SW900 upregulated the expression of GPX4, promoted proliferation, and attenuated diverse mitochondrial abnormalities specific to ferroptosis, whereas knockdown of GPX4 exacerbated such attenuations without affecting cell proliferation. Taken together, ferroptosis as an anti-tumor factor is inhibited in NSCLC, and targeting ferroptosis could be a novel therapeutic strategy for the management of NSCLC; furthermore, regulating ferroptosis could be another cancerous mechanism of STYK1.

Histone deacetylase 9 functions as an oncogene in a variety of cancers, but its role on non-small cell lung cancer (NSCLC) has not been reported. Melatonin was proven to possess anticancer actions, whereas its effect on NSCLC and underlying mechanisms remains poorly understood. In this study, 337 patients with complete clinicopathologic characteristics who underwent NSCLC surgery were recruited for the study. We found that NSCLC patients with high HDAC9 expression were correlated with worse overall survival and poor prognosis. HDAC9 knockdown significantly reduced NSCLC cell growth and induced apoptosis both in vivo and in vitro. Melatonin application also markedly inhibited cell proliferation, metastasis, and invasion and promoted apoptosis in NSCLC cells. Moreover, RNA-seq, real-time quantitative polymerase chain reaction, and western blot analyses showed that melatonin treatment decreased the HDAC9 level in NSCLC cells. A mechanistic study revealed that HDAC9 knockdown further enhanced the anticancer activities of melatonin treatment, whereas HDAC9 overexpression partially reversed the melatonin's anticancer effects. Additionally, the in vivo study found melatonin exerted anti-proliferative and pro-apoptotic effects on xenograft tumors which were also strengthened by HDAC9 knockdown. These results indicated that HDAC9 downregulation mediated the anti-NSCLC actions of melatonin, and targeting HDAC9 may be the novel therapeutic strategy for NSCLC.