Dihydroartemisinin (DHA), an active antimalaria metabolite derived from artemisinin, has received increasing attention for its anticancer activities. However, little is known about the anticancer mechanisms of DHA, although the existing data define its antimalaria effects by producing excessive reactive oxygen species (ROS). In this study, we showed that DHA effectively suppresses in vitro and in vivo tumor growth of non-small cell lung cancer (NSCLC) without perceptible toxicity on heart, liver, spleen, lung, and kidney tissues. Of note, DHA inhibited the expression of B7-H3 rather than PD-L1, whereas overexpression of B7-H3 completely rescued DHA's inhibition on cell proliferation and migration of NSCLC A549 and HCC827 cells. B7-H3 overexpression also largely inhibited DHA's induction on the apoptosis of the two cell lines. Furthermore, DHA treatment led to increased infiltration of CD8+ T Lymphocytes in the xenografts as compared with that of negative controls. Taken together, our results suggest that B7-H3 but not PD-L1 is involved in the antitumor effects of DHA in NSCLC, which may be indicative of an effective B7-H3 blockade and further combination with anti-PD-L1/PD-1 immunotherapy.

Vasculogenic mimicry (VM), an alternative microvascular circulation independent of angiogenesis, is formed by aggressive cancer cells. Tumor-expressed B7-H3 has been reported to promote VM formation in hepatocellular carcinoma and modulate angiogenesis in breast cancer and colorectal cancer. However, its effects on VM generation and angiogenesis in non-small cell Lung cancer (NSCLC) remained to be elucidated. CRISPR/Cas9-mediated B7-H3 knockout (KO) was conducted in NSCLC A549 and H3255 cells. The expression of VM-related proteins, including vascular endothelial (VE)-cadherin and matrix metalloproteinase 14 (MMP14), and the secretion of vascular endothelial growth factor (VEGF) were measured by western blotting and chemiluminescence assay in both B7-H3 KO and mock-edited A549 and H3255 cells. To examine VM formation, a three-dimensional (3D) culture model was used for B7-H3 KO and mock A549 and H3255 cells. For in vivo analysis, xenograft mice models were established using B7-H3 KO and mock-edited A549 cells, and immunohistochemical (CD31) and histochemical (periodic acid-Schiff, PAS) double staining were performed to identify VM and endothelial vessels in tumor tissues. Finally, specific signaling inhibitors were used to analyze B7-H3-induced signaling pathway responsible for VE-cadherin and MMP14 expression and VM generation. Higher expression of B7-H3 was associated with a worse prognosis and more advanced T-category in NSCLC. CRISPR/Cas9-mediated B7-H3 KO in A549 and H3255 cells led to decreased expression of VE-cadherin and MMP14; however, the secretion of VEGF by the two cell lines remained unchanged. In the 3D cell culture model, both B7-H3 KO A549 and H3255 cells showed a significant reduction in the formation of capillary-like tubular structures compared to mock-edited cells. In the in vivo xenograft model, mock-edited A549 cells formed excessive PAS+ CD31− VM channels, while B7-H3 KO restrained VM formation in the xenograft tumors. However, no significant differences were found in CD31+ endothelial vessels between xenografts formed by B7-H3 KO and mock-edited A549 cells. Finally, we analyzed the signaling pathway responsible for B7-H3-induced VM formation and found that selective inhibition of the phosphoinositide 3-kinase(PI3K)/protein kinase B (AKT) hyperactivation by LY294002 was associated with decreased expression of MMP14 and VE-cadherin, and in vitro VM formation by both A549 and H3255 cells. Tumor-expressed B7-H3 acts via PI3K/AKT signaling pathway to promote VM formation by NSCLC cells while bears no effects on angiogenesis in NSCLC.

目的 探讨双氢青蒿素(DHA)对非小细胞肺癌(NSCLC)细胞生长、迁移侵袭和血管生成拟态(VM)能力的影响.方法 将不同浓度的DHA加入NSCLC细胞株A549与H3255 中,待24h后,CCK8 法检测细胞活力,Transwell实验检测NSCLC细胞的迁移侵袭能力.qRT-PCR和 Western blot分别检测E-cadherin、N-cadherin、Vimentin mRNA和蛋白表达水平.三维细胞培养观察细胞的血管样形态生成情况.qRT-PCR和Weston blot检测VM的标志物VE-cadherin mR-NA和蛋白表达水平.结果 DHA抑制A549 和H3255 的细胞生长,并呈现时间依赖性和浓度依赖性;DHA抑制 A549和H3255 转移和侵袭能力(均P<0.001);DHA在A549 和H3255 细胞中上调E-cadherin mRNA(均P<0.001)和蛋白(P<0.001;P<0.01)的表达,下调N-cadherin mRNA(均P<0.01)和蛋白(均P<0.001)的表达以及Vimentin mRNA(P<0.01;P<0.001)和蛋白(均P<0.001)的表达.三维细胞培养结果表明,DHA处理的 A549 细胞和 H3255 细胞12hVM生成能力均降低,且下调 VE-cadherin mRNA(P<0.001;P<0.01)和蛋白(均P<0.001)表达.结论 DHA可抑制NSCLC细胞的生长、迁移侵袭和VM能力.

目的 探讨经典血管内皮生长因子A(VEGFA)和促血管拟态生成因子基质金属蛋白酶 14(MMP-14)在肺腺癌中表达及临床意义.方法 采用TCGA和UALCAN数据库分析肺腺癌MMP-14 和VEGFA基因及蛋白表达水平及其与生存预后的相关性.收集 69 例肺腺癌患者及 20 例健康受试者(对照组)血清,分别采用 ELISA 和化学发光法检测MMP-14 和VEGFA含量,分析二者与肿瘤临床病理学特征相关性及对肺腺癌的预测、诊断价值.结果 肺腺癌组织和血清中MMP-14 和VEGFA水平均高于对照组.肺腺癌早期组、晚期组和对照组血清VEGFA和MMP-14 表达水平差异均有统计学意义(P<0.001).MMP-14 在T3/T4 期高于T1/T2 分期(P = 0.045),N2/N3 期高于 N0/N1 期(P =0.035),胸膜转移组高于未转移组(P =0.034).VEGFA在M1 期高于M0 期(P =0.025).VEGFA水平升高为肺腺癌的危险因素(P =0.002).MMP-14、VEGFA和癌胚抗原(CEA)单项诊断肺腺癌的曲线下面积(AUC)分别为 0.793、0.849、0.851,三项联合检测AUC为 0.952.MMP-14、VEGFA低表达组的总生存期(OS)和疾病特异生存期(DSS)均长于 MMP-14、VEGFA高表达组.结论 MMP-14 和VEGFA在肺腺癌中高表达,分别与肺腺癌的生长、侵袭和转移有关,并对生存预后判断有一定意义.

This study aims at screening and validation of prospective genetic signature for lung adenocarcinoma (LUAD) prognosis and treatment. The immune-related genes (IRGs) were obtained from The Cancer Genome Atlas (TCGA) dataset where a total of 535 LUAD and 59 control samples were included. A risk model was then developed for the risk stratification of LUAD patients. The immune cell infiltration, clinical outcomes, and the therapeutic efficacy of programmed cell death protein 1 (PD-1) and its ligand (PD-L1) blockade were compared between high and low-risk groups. Gene set enrichment analysis (GSEA) and gene set variation analysis (GSVA) were used to explore the biological processes and signalling pathways associated with the IRGs. Finally, IRGs mRNA levels were assayed by reverse transcription quantitative real-time PCR (RT-qPCR) in LUAD and relevant cell lines. Two IRGs, P2RX1 (purinergic receptor P2X 1) and PCP4 (Purkinje cell protein 4), were screened from a module that possesses the highest correlation with plasma cells. RT-qPCR verified the expression of the two IRGs in plasmacytoma cell RPMI 8226 but not in LUAD cells. A higher risk score is associated with a lower infiltration of immune cells. Kaplan–Meier and nomogram analysis showed that the high-risk group has a lower survival rate than the low-risk cohort. Furthermore, the high-risk group had a worse response rate to PD-L1/PD-1 blockade. GSVA and GSEA-GO results indicated that a lower risk score is linked to signalling pathways and biological functions promoting immune response and inflammation. In contrast, a higher risk score is associated with signalling cascades promoting tumour growth. The immune-related prognostic model based on P2RX1 and PCP4 is conducive to predicting the therapeutic response of PD-L1/PD-1 blockade and clinical outcomes of LUAD.

BackgroundChaperonin-containing tailless complex polypeptide 1 (TCP1) subunit 3 (CCT3) has tumor-promoting effects in lung adenocarcinoma (LUAD). This study aims to investigate the molecular mechanisms of CCT3 in LUAD oncogenesis.MethodsThe UALCAN databases, Human Protein Atlas (HPA) and The Cancer Genome Atlas (TCGA) data were used to analyze CCT3 expression in LUAD. Both the Wilcoxon rank-sum test and the regression model were used to investigate the connection between clinicopathologic characteristics of LUAD patients and CCT3 expression. The prognostic value of CCT3 was determined by Cox regression models, the Kaplan-Meier method and Nomogram prediction. Next, we identified the most related genes with CCT3 via GeneMANIA and String databases, and the association between CCT3 and infiltrated immune cells using single-sample Gene Set Enrichment Analysis (ssGSEA). CCT3-related pathway enrichment analysis was investigated by GSEA. Finally, CCT3 roles in cell proliferation and apoptosis of LUAD A549 cells was verified by siRNA (small interfering RNA) mediated CCT3 knockdown.ResultsCCT3 was upregulated in LUAD both in mRNA and protein levels. CCT3 overexpression was associated with clinicopathological characteristics including sex, smoking, T- and N-categories, pathological staging, and a poor prognosis of LUAD patients. GeneMANIA and String databases found a set of CCT3-related genes that are connected to the assembly and stability of proteins involved in proteostasis of cytoskeletal filaments, DNA repair and protein methylation. Furthermore, CCT3 was found to be positively correlated with the infiltrating Th2 cells (r = 0.442, p < 0.01) while negatively correlated with mast cells (r = -0.49, p < 0.01) and immature dendritic cells (iDCs, r = -0.401, p < 0.001) according to ssGSEA analyzes. The pathway analysis based on GSEA method showed that the cell cycle pathway, the protein export pathway, the proteasome pathway and the ribosome pathway are enriched in CCT3 high group, whereas the JAK/STAT pathway, B cell receptor pathway, T cell receptor pathway and toll like receptor pathway were enriched in CCT3 low group. Finally, CCT3 knockdown substantially inhibited proliferation while promoted apoptosis of A549 cells.ConclusionIntegrated analyzes identify CCT3 as a modulator to shape immunosuppressive tumor microenvironment in LUAD and therefore, a prognostic factor for LUAD.

目的 探讨肺腺癌表皮生长因子受体(EGFR)基因突变与脑转移的相关性.方法 比较EGFR 19 外显子缺失(19 Del)和21 外显子点突变(21 L858R)患者初诊以及接受表皮生长因子受体酪氨酸激酶抑制剂(EGFR-TKIs)靶向治疗后脑转移情况;CCK-8 法检测并计算EGFR突变肺腺癌细胞株PC9(19 Del)和H3255(21 L858R)对吉非替尼的半抑制浓度(IC50).结果 纳入410 例肺腺癌患者,总体脑转移153 例(37.3%),年龄、淋巴结转移和EGFR 21 L858R突变是脑转移高危因素.其中初诊EGFR突变患者脑转移 48例,19 Del和21 L858R分别为 14 和 34 例(P =0.006);EG-FR-TKIs靶向治疗后有17 例 19 Del和 20 例 21 L858R患者发生脑转移,其脑转移中位数时间(月)分别为 15(8.50,25.00)和7(4.25,12.25)(P =0.013).PC9 和H3255 对吉非替尼的 IC50 分别(0.037±0.008)和(0.150±0.040)μmol/L(P =0.007).结论 年龄≤60 岁、淋巴结N2～3 分期及EGFR 21 L858R突变是肺腺癌患者发生脑转移的高危因素,后者在EGFR-TKIs治疗后发生脑转移时间仍短于 19 Del突变,可能与药物敏感性有关.

Backgrounds & aims Epstein-Barr virus (EBV) infection occurs commonly in children and may cause acute infectious mononucleosis (AIM) and various malignant diseases. Host immune responses are key players in the resistance to EBV infection. We here assessed the immunological events and laboratory indicators of EBV infection, as well as determined the clinical usefulness of evaluating the severity and efficacy of antiviral therapy in AIM patients. Methods We enrolled 88 children with EBV infection. The immune environment was defined by immunological events such as frequencies of lymphocyte subsets, phenotypes of T cells, and their ability to secrete cytokines, and so on. This environment was analyzed in EBV-infected children with different viral loads and in children in different phases of infectious mononucleosis (IM) from disease onset to convalescence. Results Children with AIM had higher frequencies of CD3 + T and CD8 + T cells, but lower frequencies of CD4 + T cells and CD19 + B cells. In these children, the expression of CD62L was lower and that of CTLA-4 and PD-1 was higher on T cells. EBV exposure induced granzyme B expression, but reduced IFN- γ secretion, by CD8 + T cells, whereas NK cells exhibited reduced granzyme B expression and increased IFN- γ secretion. The frequency of CD8 + T cells was positively correlated with the EBV DNA load, whereas the frequencies of CD4 + T cells and B cells were negatively correlated. During the convalescent phase of IM, CD8 + T cell frequency and CD62L expression on T cells were restored. Moreover, patient serum levels of IL-4, IL-6, IL-10, and IFN- γ were considerably lower throughout the convalescent phase than throughout the acute phase. Conclusion Robust expansion of CD8 + T cells, accompanied by CD62L downregulation, PD-1 and CTLA-4 upregulation on T cells, enhanced granzyme B production, and impaired IFN- γ secretion, is a typical characteristic of immunological events in children with AIM. Noncytolytic and cytolytic effector functions of CD8 + T cells are regulated in an oscillatory manner. Furthermore, the AST level, number of CD8 + T cells, and CD62L expression on T cells may act as markers related to IM severity and the effectiveness of antiviral treatment.

COVID-19 pandemic has already had a tremendous impact on the process of human society; the survival of mankind and the healthy living environment deterioration with the influence will last for many years. This meta-analysis aims to assess the risk of COVID-19 in patients with rheumatic diseases. PubMed, Web of Science, Embase, China National Knowledge Infrastructure (CNKI), and Chinese Biomedical Database (CBM) were systematically searched with no language restriction up to July 5, 2021. The pooled rates were synthesized by fixed effect model or random effect model depending on heterogeneity. A total of 83 articles were included in this meta-analysis. The incidence of COVID-19 in patient with rheumatic diseases was 0.0190 (95% CI: 0.0136-0.0252), and the hospitalization rate, intensive care unit admission rate, mechanical ventilation rate, and case fatality rate of patients with rheumatic diseases infected with COVID-19 were 0.4396 (95% CI: 0.3899-0.4898), 0.0635 (95% CI: 0.0453-0.0836), 0.0461 (95% CI: 0.0330-0.0609), and 0.0346 (95% CI: 0.0218-0.0493), respectively. Our research shows that patients with rheumatic diseases have great risk of COVID-19. Differences in COVID-19 incidence, hospitalization rates, and mortality rates in regions were statistically significant. We still need to pay attention to the risk of COVID-19 in patients with rheumatic diseases. • Although the risk of COVID-19 in patients with rheumatic diseases has been discussed in previous meta-analysis, their research directions were inconsistent, and few studies focus on prevalence or serious outcomes of COVID-19 in patient with rheumatic diseases, while the quality of these articles was variable. • The incidence of COVID-19 and serious clinical outcomes in patients with rheumatic diseases were still high along with differential risks in most regions. • The use of glucocorticoids and conventional synthetic disease-modifying antirheumatic drugs did not affect the hospitalization rate and mortality in rheumatism patients with COVID-19.

Background: Hepcidin is the master iron regulator hormone produced by the liver. The association of serum hepcidin with pegylated interferon therapy in patients with chronic hepatitis C infection has been studied. However, the role of serum hepcidin level in predicting the effect of pegylated interferon treatment in patients with chronic hepatitis B (CHB) infection is yet to be elucidated. Our study aims to investigate the correlation between hepcidin expression levels and the curative effect of interferon-alpha therapy in patients with CHB. Methods: A total of 47 patients with CHB who accepted pegylated interferon-alpha (PEG-IFN-alpha) treatment were recruited. The serum level of hepcidin was estimated by ELISA. The alternation in the gene expression level of hepcidin was detected by RT-PCR, and immunofluorescence cell staining was performed to detect hepcidin peptide. The induction of antiviral proteins was analyzed by Western blotting. The predictive value of early on-treatment variation in serum hepcidin during treatment progress was assessed by receiver operating charac-teristic analysis. Results: High levels of early on-treatment serum hepcidin were observed in patients who achieved a decline in HBsAg > 1 log10 IU/mL or HBV DNA > 1 log10 IU/mL. In vitro, an elevation of the hepcidin expression in HepG2.2.15 cells induced by PEG-IFN-alpha treatment was noted. Furthermore, combined treatment with hepcidin and PEG-IFN-alpha increased the levels of antiviral proteins. The predictive cut-off value of hepcidin for HBsAg decline > 1 log10 IU/mL was 239 pg/mL, and the sensitivity and specificity were 72.73% and 70.97%, respectively. The predictive cut-off value of hepcidin for the decline in HBV DNA > 1 log10 IU/mL was 190.4 pg/ mL, and the sensitivity and specificity were 72.73% and 61.11%, respectively. Early-on treatment changes in the hepcidin level signified the predictive value of the PEG-IFN-alpha curative effect. Conclusions: A higher early-on treatment hepcidin level indicates a higher possibility of HBsAg and HBV DNA decline in patients with CHB during PEG-IFN-alpha treatment. A high early-on treatment serum hepcidin level is significant in predicting the PEG-IFN-alpha therapeutic effect in patients with CHB.

Abstract Background Not all lung adenocarcinoma (LUAD) patients with activating epidermal growth factor receptor (EGFR) mutations respond to tyrosine kinase inhibitors (TKIs) as intended. Thus, biomarkers are needed to identify patients who benefit most from EGFR-targeted therapy. Our previous in vitro data has shown that the co-signal molecule B7-H3 determines EGFR-TKI gefitinib susceptibility of EGFR-mutated LUAD cell lines, based on the potential crosslinking between B7-H3-induced signaling and EGFR signaling. Methods We detected tumoral B7-H3 expression in the original biopsy from 56 treatment-naïve LUAD patients and analyzed the association between high/low B7-H3 expression with the clinical outcomes of first-line anti-EGFR therapy. The main criteria for the analysis of response were overall response rate (ORR), disease control rate (DCR), and progression-free survival (PFS), and the secondary criterion was overall survival (OS). Results In the subgroups of B7-H3 high and low expression, the ORR were 16.0% (4/25) and 74.2% (23/31) (p<0.001), and the DCR were 36.0% (9/25) and 87.1% (27/31) (p<0.001), respectively. The PFS of B7-H3 high [median 8.7, 95% confidence interval (CI) 4.0–13.4] was significantly worse than that of B7-H3 low (median not reached) [HR 6.54 (95% CI 2.18–19.60), p=0.001]. The median OS was 15.9 (95% CI 10.0–21.8) months in the B7-H3 high cohort and 25.7 (95% CI 9.0–42.4) months in the B7-H3 low subjects [HR 2.08 (95% CI 1.07–4.02), p=0.03], respectively. Both the univariate and multivariate analyses identified B7-H3 as an independent factor associated with poor PFS (p=0.001, p=0.000) and OS (p=0.03, p=0.015). Conclusion B7-H3 may serve as a potential biomarker to predict clinical outcomes in EGFR-mutated LUAD patients treated with first-line EGFR-TKIs.

Endoplasmic reticulum aminopeptidase 1 (ERAP1) is a processing enzyme of antigenic peptides presented to major histocompatibility complex (MHC) class I molecules. ERAP1-dependent trimming of epitope repertoire determines an efficacy of adoptive CD8+ T-cell responses in several viral diseases; however, its role in hepatitis B virus (HBV) infection remains unknown. Here, we show that the serum level of ERAP1 in patients with chronic hepatitis B (CHB) (n = 128) was significantly higher than that of healthy controls (n = 44) (8.78 ± 1.82 vs. 3.52 ± 1.61, p < 0.001). Furthermore, peripheral ERAP1 level is moderately correlated with HBV DNA level in patients with CHB (r = 0.731, p < 0.001). HBV-transfected HepG2.2.15 cells had substantially increased ERAP1 expression and secretion than the germline HepG2 cells (p < 0.001). The co-culture of ERAP1-specific inhibitor ERAP1-IN-1 pretreated HepG2.2.15 cells or ERAP1 knockdown HepG2.2.15 cells with CD8+ T cells led to 14–24% inhibition of the proliferation of CD8+ T cells. Finally, liquid chromatography tandem mass spectrometry (LC-MS/MS) test demonstrated that ERAP1-IN-1 blocks completely the production of a 9-mers peptide (30–38, LLDTASALY) derived from Hepatitis B core antigen (HBcAg). The predictive analysis by NetMHCpan-4.1 server showed that human leukocyte antigen (HLA)-C*04:01 is a strong binder for the 9-mers peptide in HepG2.2.15 cells. Taken together, our results demonstrated that ERAP1 trims HBcAg to produce 9-mers LLDTASALY peptides for binding onto HLA-C*04:01 in HepG2.2.15 cells, facilitating the potential activation of CD8+ T cells.

目的 探讨乙型肝炎病毒(HBV)感染所致的第10号染色体缺失的磷酸酶及张力蛋白同源的基因(PTEN)下调与α-干扰素(IFN-α)抗病毒活性的相关机制.方法 在适宜条件下培养HepG2细胞和HepG2.2.15细胞,以空白载体(pcDNA3.1)和HBV1.3质粒分别转染HepG2细胞,Western blot法检测PTEN蛋白质的表达;将pcDNA3.1和PTEN过表达(PTEN-OE)质粒分别瞬时转染HepG2.2.15细胞,化学发光法分析细胞培养上清液中HBV相关抗原的表达,实时荧光定量PCR(qRT-PCR)技术分析HBV前基因组RNA(HBV pgRNA)表达;使用合成RNA双链体poly(I:C)刺激PTEN-OE的细胞,qRT-PCR技术分析IFN-αmRNA的表达,West-ern blot法分析JAK/STAT信号通路中干扰素调节因子9(IRF9)、黏病毒抗性蛋白1(MxA)的表达.结果 瞬时表达HBV的HepG2细胞和稳定表达HBV的HepG2.2.15细胞中,PTEN蛋白的表达降低;PTEN-OE的HepG2.2.15细胞中,HBV相关抗原及HBV pgRNA的表达较对照组降低,经poly(I:C)作用后,IFN-αmRNA水平较对照组显著升高,且JAK/STAT信号通路相关蛋白IRF9、MxA的表达增加.结论 HBV可能通过下调PTEN的表达发挥拮抗IFN-α抗病毒活性的作用.

目的 探讨共信号分子B7-H3与肺腺癌表皮生长因子受体酪氨酸激酶抑制剂(EGFR-TKI)靶向治疗继发性耐药的相关性.方法 采用免疫组化方法检测56例肺腺癌组织B7-H3表达,将其分为低表达和高表达两组,比较两组患者EGFR-TKI靶向治疗继发性耐药发生比例和发生时间;体外诱导培养吉非替尼耐药的EGFR突变肺腺癌细胞株H3255和HCC827,以野生型细胞株作为对照,流式细胞术分析耐药株B7-H3表达水平.结果 56例肺腺癌患者中B7-H3低表达31例,高表达25例;截止该研究观察终止时间,B7-H3低表达和高表达患者继发性耐药发生例数分别为12例(38.7％)和14例(56.0％)(P=0.19,χ2=1.663),其中位耐药时间分别为13.2个月和5.1个月(P=0.004,t=3.193);与野生型H3255和HCC827细胞株比较,耐药株B7-H3表达水平进一步增加.结论 B7-H3表达水平与肺腺癌EGFR-TKI靶向治疗继发性耐药存在相关性.

Epithelial-mesenchymal transition (EMT) is a key step in cancer metastasis. B7-H3, a co-signaling molecule associated with poor prognosis of non-small cell lung cancer (NSCLC), promotes the metastasis of NSCLC by activating the EMT process. However, its underlying mechanism remains poorly understood. In the present study, it was shown that CRISPR/Cas9-mediated B7-H3 deletion downregulated the expression of the class III histone deacetylase, sirtuin-1 (SIRT1), in NSCLC A549 cells. Accordingly, SIRT1 silencing resulted in markedly decreased migration and invasion of A549 cells. Both B7-H3 gene-edited and SIRT1-silenced cells were typically characterized by an increased expression of the epithelial marker E-cadherin, and downregulation of the mesenchymal markers N-cadherin and vimentin, as compared with mock-edited and scrambled negative small interfering RNA control, respectively. It was further demonstrated that B7-H3 ablation significantly downregulated phosphorylated AKT/protein kinase B expression, and SIRT1 expression was substantially suppressed by the PI3K-specific inhibitor, LY294002. Taken together, the findings of the present study revealed that B7-H3-induced signaling upregulates SIRT1 expression via the PI3K/AKT pathway to promote EMT activation that is associated with metastasis in NSCLC.

INTRODUCTION:The introduction of antiviral therapy in chronic hepatitis B (CHB) infection depends on precise evaluation of hepatic lesions. Total serum bile acids (TSBAs) are highly sensitive in monitoring liver dysfunction. We evaluated the predictive role of TSBAs for hepatic lesions in CHB patients with borderline alanine aminotransferase (ALT) and high level of hepatitis B virus (HBV) DNA copies.METHODOLOGY:328 CHB patients were enrolled, 241 were hepatitis B e antigen (HBeAg)-positive and 87 were HBeAg-negative. Patients were further divided into two entities according to inflammation/fibrosis evaluated by liver biopsy, low-grade (inflammation grade < 2 and fibrosis stage < 2) and high-grade (inflammation grade ≥ 2 or/and fibrosis stage ≥ 2) cohorts. TSBAs were compared with noninvasive tools including aspartate aminnotransferase (AST)-to-platelet ratio index (APRI), fibrosis-4 (FIB-4) and red cell distribution width (RDW)-to-platelet ratio (RPR) to predict high-grade hepatic lesions in CHB subgroups.RESULTS:TSBAs, APRI, FIB-4 and RPR were statistically different between low- and high-grade patients in HBeAg-positive cohort. Only TSBAs showed significant difference between low and high grade in HBeAg-negative patients. Similarly, APRI, FIB-4 and RPR were correlated with different division of inflammation/fibrosis only in HBeAg-positive while TSBAs were correlated with inflammation/fibrosis levels in both HBeAg-positive and HBeAg-negative groups. Of the four indicators, the receiver operating characteristic (ROC) curve analysis showed that TSBAs have the maximum AUC (area under the curve) in HBeAg-negative group but the minimum in HBeAg-positive cohort.CONCLUSIONS:TSBAs can be used for predicting antiviral therapy in CHB patients with HBeAg-negative, borderline ALT and high HBV DNA.

目的 探讨外周血内质网氨肽酶-1(ERAP1)水平对肝细胞肝癌(HCC)病情严重程度和预后的诊断价值.方法 采用酶联免疫吸附实验(ELISA)检测HCC患者、肝硬化患者及健康对照(HC)血清ERAP1水平;分析ERAP1和临床各项指标的相关性,采用多因素Logistic回归分析病情程度和预后的独立危险因素.受试者工作曲线(ROC)评估ERAP1在诊断不同病情程度和预后的敏感性和特异性.结果 HCC患者血清ERAP1水平与HC组、肝硬化组比较,差异有统计学意义(P<0.05),ERAP1水平与肿瘤分期、大小及病灶数目均呈显著相关性(均P<0.05);ERAP1水平与丙氨酸氨基转移酶(ALT)、血清天冬氨酸氨基转移酶(AST)、血清总胆红素(TBIL)、甲胎蛋白(AFP)均呈正相关,与ALB呈负相关(P<0.05).多因素Logistic回归分析显示ERAP1为病情程度和不同预后的独立预测因素.ERAP1和AFP联合诊断早期HCC的曲线下面积(AUC)为0.932;诊断不良预后时AUC为0.742.结论 ERAP1水平在评估HCC患者病情程度和预后方面具有重要的临床意义和潜在应用价值.

Abstract Background Chaperonin-containing tailless complex polypeptide 1 (TCP1) subunit 3 (CCT3) is overexpressed in a wide range of cancers and has a connection with poor prognosis. However, its role in lung adenocarcinoma (LUAD), the most common type of non-small cell lung cancer (NSCLC) remained to be elucidated. Methods The Timer and UALCAN databases, Human Protein Atlas (HPA) and The Cancer Genome Atlas (TCGA) data were used to analyze CCT3 expression in LUAD. Both the Wilcoxon rank-sum test and the regression model were used to investigate the connection between clinicopathologic characteristics of LUAD patients and CCT3 expression. The diagnostic efficiency of CCT3 for LUAD was assessed by receiver operating curve, and the prognostic value of CCT3 was determined by Cox regression models, the Kaplan-Meier method and Nomogram prediction. Next, we identified the most related genes with CCT3 via GeneMANIA and String databases, and the association between CCT3 and infiltrated immune cells with single-sample Gene Set Enrichment Analysis (ssGSEA). CCT3-related infiltrating immune cells was investigated by GSEA. Finally, CCT3 roles in cell proliferation and apoptosis of LUAD A549 cells was verified by siRNA (small interfering RNA) mediated knockdown. Results CCT3 was upregulated in LUAD both in mRNA and protein levels. CCT3 overexpression was associated with clinicopathological characteristics including sex, smoking, T- and N-categories, pathological staging, and a poor prognosis of LUAD patients. GeneMANIA and String databases found a set of CCT3-related genes that are connected to the assembly and stability of proteins involved in proteostasis of cytoskeletal filaments, DNA repair and oncogenesis. Furthermore, CCT3 was found to be positively correlated with the infiltrating Th2 cells (r = 0.442, p < 0.01) while negatively correlated with mast cells (r = -0.49, p < 0.01) and immature dendritic cells (iDCs, r = -0.401, p < 0.001) according to ssGSEA analyzes. The pathway analysis based on GSEA method showed that the cell cycle pathway, the protein export pathway, the proteasome pathway and the ribosome pathway are enriched in CCT3 high group, whereas the JAK/STAT pathway, B cell receptor pathway, T cell receptor pathway and toll like receptor pathway were enriched in CCT3 low group. Finally, CCT3 knockdown substantially inhibited proliferation while promoted apoptosis of A549 cells. Conclusion Dysregulation of CCT3 in LUAD is associated with unique genetic and immune properties and has excellent diagnostic and prognostic performance for LUAD.

Introduction: The introduction of antiviral therapy in chronic hepatitis B (CHB) infection depends on precise evaluation of hepatic lesions. Total serum bile acids (TSBAs) are highly sensitive in monitoring liver dysfunction. We evaluated the predictive role of TSBAs for hepatic lesions in CHB patients with borderline alanine aminotransferase (ALT) and high level of hepatitis B virus (HBV) DNA copies. Methodology: 328 CHB patients were enrolled, 241 were hepatitis B e antigen (HBeAg)-positive and 87 were HBeAg-negative. Patients were further divided into two entities according to inflammation/fibrosis evaluated by liver biopsy, low-grade (inflammation grade < 2 and fibrosis stage < 2) and high-grade (inflammation grade ≥ 2 or/and fibrosis stage ≥ 2) cohorts. TSBAs were compared with noninvasive tools including aspartate aminnotransferase (AST)-to-platelet ratio index (APRI), fibrosis-4 (FIB-4) and red cell distribution width (RDW)-to-platelet ratio (RPR) to predict high-grade hepatic lesions in CHB subgroups. Results: TSBAs, APRI, FIB-4 and RPR were statistically different between low- and high-grade patients in HBeAg-positive cohort. Only TSBAs showed significant difference between low and high grade in HBeAg-negative patients. Similarly, APRI, FIB-4 and RPR were correlated with different division of inflammation/fibrosis only in HBeAg-positive while TSBAs were correlated with inflammation/fibrosis levels in both HBeAg-positive and HBeAg-negative groups. Of the four indicators, the receiver operating characteristic (ROC) curve analysis showed that TSBAs have the maximum AUC (area under the curve) in HBeAg-negative group but the minimum in HBeAg-positive cohort. Conclusions: TSBAs can be used for predicting antiviral therapy in CHB patients with HBeAg-negative, borderline ALT and high HBV DNA.