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Dysregulated cholesterol metabolism is a hallmark of colorectal cancer (CRC). However, the usage of cholesterol-lowering agents seemed to have no benefit in CRC patients. In this study, we focused on the cholesterol-nuclear receptors (NRs) axis as a strategy. Cholesterol and its derivatives work as ligands for different nuclear receptors, thus promoting cancer progression. The key NR downstream of cholesterol in CRC is unknown. Here, we treated CRC cells with a cholesterol-lowering agent and lipoprotein-depleted conditioned medium, and then detected the change of the putative NRs. The results revealed that RORα/γ (Retinoic acid receptor-related Orphan Receptor α/γ) levels exhibited the most obvious increases in CRC cells subjected them to cholesterol deprivation. RORα/γ agonists significantly inhibited CRC cells proliferation and migration in vitro and in vivo. Also, RORα/γ overexpression repressed CRC cells proliferation and migration in vitro and in vivo and RORα/γ knockdown promoted it. Mechanistically, RORα/γ agonists promoted c-myc degradation by activating the transcription of the ubiquitinase NEDD4. Intriguingly, the combination of RORα/γ agonists and atorvastatin had a synergistic effect on inhibiting CRC cells. These findings demonstrate that the cholesterol- RORα/γ axis is important for maintaining c-myc protein levels. Combination therapy with atorvastatin and RORα/γ agonist is a promising therapeutic strategy for CRC.

Background Due to its limited efficacy and potential toxicity, anti-PD-1 monoclonal antibody is not suitable for all advanced gastric cancer (AGC) patients and predictive biomarkers identifying patients who can benefit from it are urgently needed. This study aimed to evaluate the predictive and prognostic value of inflammatory markers in the context of the systemic inflammatory status and tumour microenvironment. Methods The study included 58 patients from a prospective study investigating the safety and efficacy of toripalimab in chemorefractory AGC patients. Patient characteristics, treatment outcomes, and haematological parameters were analysed. Immune-cell infiltration and gene expression in tumour tissue were examined using transcriptome sequencing. Results In this cohort, the median follow-up time was 4.5 months, the median progression-free survival was 1.9 months, and the median overall survival (OS) was 4.8 months. The objective response rate was 12.1% and th disease control rate (DCR) was 39.7%. Both the baseline blood neutrophil-to-lymphocyte ratio (bNLR) with a cut-point of 2.7 and the early elevated dynamic change of the bNLR (dNLR) with a cut-point of 1.5 were prognostic factors of survival. Patients in the high bNLR or dNLR group had remarkably poor DCR (25.8% vs 59.1%, P = 0.023; 15.8% vs 54.6%, P = 0.008). In multivariate analysis, bNLR and tumour mutational burden were independent prognostic factors of OS. Tumour RNA-seq analysis revealed enriched neutrophil infiltration and a higher tumour NLR in the bNLR-high group. Corresponding tumour gene-expression profiles were associated with neutrophil recruitment and inflammatory cytokine aggregation. Conclusions Our study demonstrated the potential clinical utility of NLR as a biomarker for patient selection and clinical management in predicting the prognosis of AGC patients as well as response to anti-PD-1 therapy. In addition, high bNLR reflected the imbalance of tumour-tissue-infiltrating neutrophils and lymphocytes, and was associated with an immunosuppressive and pro-tumour microenvironment.

Fat mass and obesity-associated protein (FTO), an N6-methyladenosine (m6A) demethylase, participates in tumor progression and metastasis in many malignancies, but its role in colorectal cancer (CRC) is still unclear. Here, we found that FTO protein levels, but not RNA levels, were downregulated in CRC tissues. Reduced FTO protein expression was correlated with a high recurrence rate and poor prognosis in resectable CRC patients. Moreover, we demonstrated that hypoxia restrained FTO protein expression, mainly due to an increase in ubiquitin-mediated protein degradation. The serine/threonine kinase receptor associated protein (STRAP) might served as the E3 ligase and K216 was the major ubiquitination site responsible for hypoxia-induced FTO degradation. FTO inhibited CRC metastasis both in vitro and in vivo. Mechanistically, FTO exerted a tumor suppressive role by inhibiting metastasis-associated protein 1 (MTA1) expression in an m6A-dependent manner. Methylated MTA1 transcripts were recognized by an m6A “reader”, insulin-like growth factor 2 mRNA binding protein 2 (IGF2BP2), which then stabilized its mRNA. Together, our findings highlight the critical role of FTO in CRC metastasis and reveal a novel epigenetic mechanism by which the hypoxic tumor microenvironment promotes CRC metastasis.

Fluxametamide is a novel isoxazoline insecticide and has been registered in Korea and Japan to control Lepidoptera pests. Rice stem borer, Chilo suppressalis (Walker), is a destructive Lepidoptera pest of rice in China, and novel effectively insecticides are required to be developed for controlling it due to its increasing resistance levels. Therefore, the lethal and sublethal effects of fluxametamide on C. suppressalis were investigated in the present study. In the lethal assay, the insecticidal activity of fluxametamide with median lethal dose (LD50) value of 1.308 mg/kg to the 4th instar larvae of C. suppressalis was higher than that of chlorantraniliprole (LD50, 3.112 mg/kg) and lower than that of emamectin benzoate (LD50, 0.006 mg/kg). In addition, the 3rd instar larvae of C. suppressalis were sensitive to fluxametamide than the 4th instar larvae. In the sublethal (LD10 and LD30) assay, the duration of 3rd to 6th instar larvae was significantly increased, whereas the pupal duration, pupation rate, and life-cycle rate were also significantly increased in F0 generation. Both length and weight of ovarian tube were decreased with the increase of fluxametamide dose, and they in the LD30 treatment were significantly lower than those of the control group. In F1 generation, only the duration of eggs was significantly increased with LD30 treatment of fluxametamide, other developmental parameters has no significant change. These results suggest that fluxametamide has excellent lethal and sublethal effects on C. suppressalis and probably is able to suppress the population growth and progeny of C. suppressalis.

Background Numerous reports on microRNAs have illustrated their role in tumor growth and metastasis. Recently, a new prognostic factor, miR‐125b‐2‐3p, has been identified for predicting chemotherapeutic sensitivity in advanced colorectal cancer (CRC). However, the specific mechanisms and biological functions of miR‐125b‐2‐3p in advanced CRC under chemotherapy have yet to be elucidated. Methods MiR‐125b‐2‐3p expression was detected by real‐time PCR (RT‐PCR) in CRC tissues. The effects of miR‐125b‐2‐3p on the growth, metastasis, and drug sensitivity of CRC cells were tested in vitro and in vivo. Based on multiple databases, the upstream competitive endogenous RNAs (ceRNAs) and the downstream genes for miR‐125b‐2‐3p were predicted by bioinformatic analysis, followed by the experiments including luciferase reporter assays, western blot assays, and so on. Results MiR‐125b‐2‐3p was significantly lowly expressed in the tissues and cell lines of CRC. Higher expression of miR‐125b‐2‐3p was associated with relatively lower proliferation rates and fewer metastases. Moreover, overexpressed miR‐125b‐2‐3p remarkably improved chemotherapeutic sensitivity of CRC in vivo and in vitro. Mechanistically, miR‐125b‐2‐3p was absorbed by long noncoding RNA (lncRNA) XIST regulating WEE1 G2 checkpoint kinase (WEE1) expression. The upregulation of miR‐125b‐2‐3p inhibited the proliferation and epithelial‐mesenchymal transition (EMT) of CRC induced by lncRNA XIST. Conclusions Lower miR‐125b‐2‐3p expression resulted in lower sensitivity of CRC to chemotherapy and was correlated with poorer survival of CRC patients. LncRNA XIST promoted CRC metastasis acting as a ceRNA for miR‐125b‐2‐3p to mediate WEE1 expression. LncRNA XIST‐miR‐125b‐2‐3p‐WEE1 axis not only regulated CRC growth and metastasis but also contributed to chemotherapeutic resistance to CRC., Low expression of miR‐125b‐2‐3p in CRC was linked to lower chemotherapeutic sensitivity and poor survival. The lncRNA XIST promoted CRC invasion and migration by functioning as a ceRNA for miR‐125b‐2‐3p to mediate WEE1 expression. Our findings suggest that the lncRNA XIST‐miR‐125b‐2‐3p‐WEE1 axis not only regulated CRC growth and metastasis but also contributed to chemotherapeutic resistance to CRC, which may shed light on their targeted applications.

Pre-stressed bolt anchorage is the key technology for jointed rock masses in rock tunnelling, slope treatment and mining engineering. To investigate the mechanical properties and reinforcement effect of jointed rock masses with pre-stressed bolts, in this study, uniaxial compression tests were conducted on specimens with different anchoring types and flaw inclination angles. ABAQUS software was used to verify and supplement the laboratory tests. The laws of the uniaxial compressive strength (UCS) obtained from the numerical simulations and laboratory tests were consistent. The results showed that under the same flaw angle, both the UCS and elastic modulus of the bolted specimens were improved compared with those of the specimens without bolts and the improvements increased with an increase in the bolt pre-stress. Under the same anchoring type, the UCS and elastic modulus of the jointed specimens increased with an increase in the flaw angle. The pre-stressed bolt could not only restrain the slip of the specimens along the flaw surface but also change the propagation mode of the secondary cracks and limit the initiation of cracks. In addition, the plot contours of the maximum principal strain and the Tresca stress of the numerical models were influenced by the anchoring type, flaw angle, anchoring angle and bolt position.

Background Inherited susceptibility accounts for nearly one‐third of colorectal cancer (CRC) predispositions and has an 80%‐100% lifetime risk of this disease. However, there are few data about germline mutations of hereditary CRC‐related genes in Chinese patients with CRC. This study aimed to assess the prevalence of gene mutations related to cancer susceptibility among Chinese patients with CRC, differences between Chinese and Western patients, and the phenotype‐genotype correlation. Methods We retrospectively collected tumor samples from 526 patients with CRC under 70 years old who underwent hereditary CRC genetic testing. A series of bioinformatic analyses, as well as statistical comparisons, were performed. Results We found that 77 patients (14.6%) harbored functional variants of the 12 genes. The mutation frequencies of the top 5 mutated genes were 6.5% for MutL homolog 1 (MLH1), 5.1% for MutS homolog 2 (MSH2), 1.0% for MSH6, 0.8% for PMS1 homolog 2 (PMS2), and 0.8% for APC regulator of the WNT signaling pathway (APC). Our data showed much higher rates of mutations of MSH6 and PMS2 genes among all mismatch repair (MMR) genes as compared with those in Western populations. Mutations in MLH1, MSH2, and MSH6 were found to be mutually exclusive. Patients with MLH1 or MSH2 mutations had higher frequencies of personal history of cancer (MLH1: 20.6% vs. 8.7%; MSH2: 25.9% vs. 8.6%) and family history of cancer than those without these mutations (MLH1: 73.5% vs. 48.4%; MSH2: 70.4% vs. 48.9%), and the lesions were more prone to occur on the right side of the colon than on the left side (MLH1: 73.5% vs. 29.3%; MSH2: 56.0% vs. 31.0%). The proportion of stage I/II disease was higher in patients with MLH1 mutations than in those without MLH1 mutations (70.6% vs. 50.7%), and the rate of polyps was higher in patients with APC mutations than in those with wild‐type APC (75.0% vs. 17.4%). Conclusion These results provide a full‐scale landscape of hereditary susceptibility over 12 related genes in CRC patients and suggest that a comprehensive multi‐gene panel testing for hereditary CRC predisposition could be a helpful analysis in clinical practice.

Mechanical properties of the interface between a pile and soil greatly affect the bearing capacity of a pile. The creep of soil causes a long-term strength of the pile–soil interface to be smaller than its instantaneous shear strength. It is helpful to understand the settlement deformation and strength characteristics of friction piles from the micromechanism by studying the shear creep mechanical properties of the clay–concrete interface. In this work, we carry out shear creep tests on the interfaces between clay and concrete slabs with different roughnesses under different normal stresses. We analyze the influence of the joint roughness coefficient (JRC) on the long-term cohesion and long-term friction angle of the interface. We propose an instantaneous damage factor of the shear modulus and improve the Nishihara creep model with the instantaneous damage effect of the shear modulus. The experimental results show: (1) the whole process test curve of the shear creep shows that the specimen is rapidly destroyed after the transient creep stage and the steady-state creep stage, but the tertiary creep stage is not obvious. (2) The long-term shear strength of the clay-concrete interface is greatly affected by the change in the normal stress under the same JRC conditions, and the long-term shear strength of the interface increases linearly with increasing normal stress. (3) The relationship between the long-term shear strength and normal stress of the interface can be described by the Mohr–Coulomb failure criterion, and the long-term cohesion and long-term friction angle under different JRC conditions can be calculated accordingly. There exists a critical value JRCcr for the long-term cohesion, and the long-term cohesion of the interface first increases and then decreases as the JRC increases. When the long-term cohesion of the interface decreases to a certain extent, it fluctuates around the long-term cohesion value of clay. Moreover, the long-term friction angle of the interface fluctuates slightly with varying JRC, and there is no obvious change rule. (4) Considering the instantaneous damage of the shear modulus, we establish a relationship between the instantaneous shear modulus and the damage factor and introduce the damage factor into the Nishihara creep model. The shear creep mechanical parameters of the clay–concrete specimens under multistage loading are inversed.

Gastrointestinal cancer causes countless deaths every year due to therapeutic resistance. However, whether metabolic alterations contribute to chemoresistance is not well understood. In this study, we report that fatty acid (FA) catabolism was activated in gastrointestinal cancer cells treated with oxaliplatin, which exhibited higher expression of the rate-limiting enzymes carnitine palmitoyltransferase 1B (CPT1B) and CPT2. The clinical analysis also showed that high expression of these enzymes was associated with poor oxaliplatin-based chemotherapy outcomes in patients. Furthermore, genetic or pharmacological inhibition of CPT2 with perhexiline disturbed NADPH and redox homeostasis and increased reactive oxygen species (ROS) generation and cell apoptosis in gastrointestinal cancer cells following oxaliplatin treatment. Specifically, the combination of oxaliplatin and perhexiline significantly suppressed the progression of gastrointestinal cancer in cell-based xenograft and patient-derived xenograft (PDX) models. Mechanistically, CPT2 was transcriptionally upregulated by nuclear factor of activated T cells 3 (NFATc3), which translocated to the nucleus in response to oxaliplatin treatment. In summary, our study suggests that the inhibition of CPT-mediated FA catabolism combined with conventional chemotherapy is a promising therapeutic strategy for patients with gastrointestinal cancers.

BackgroundLong noncoding RNAs (lncRNAs) play nonnegligible roles in the epigenetic regulation of cancer cells. This study aimed to identify a specific lncRNA that promotes the colorectal cancer (CRC) progression and could be a potential therapeutic target.MethodsWe screened highly expressed lncRNAs in human CRC samples compared with their matched adjacent normal tissues. The proteins that interact withLINRIS(Long Intergenic Noncoding RNA for IGF2BP2 Stability) were confirmed by RNA pull-down and RNA immunoprecipitation (RIP) assays. The proliferation and metabolic alteration of CRC cells withLINRISinhibited were tested in vitro and in vivo.ResultsLINRISwas upregulated in CRC tissues from patients with poor overall survival (OS), andLINRISinhibition led to the impaired CRC cell line growth. Moreover, knockdown ofLINRISresulted in a decreased level of insulin-like growth factor 2 mRNA-binding protein 2 (IGF2BP2), a newly found N6-methyladenosine (m6A) ‘reader’.LINRISblocked K139 ubiquitination of IGF2BP2, maintaining its stability. This process prevented the degradation of IGF2BP2 through the autophagy-lysosome pathway (ALP). Therefore, knockdown ofLINRISattenuated the downstream effects of IGF2BP2, especially MYC-mediated glycolysis in CRC cells. In addition, the transcription ofLINRIScould be inhibited by GATA3 in CRC cells. In vivo experiments showed that the inhibition ofLINRISsuppressed the proliferation of tumors in orthotopic models and in patient-derived xenograft (PDX) models.ConclusionLINRISis an independent prognostic biomarker for CRC. TheLINRIS-IGF2BP2-MYC axis promotes the progression of CRC and is a promising therapeutic target.

We study the balance problem caused by forward leaning of the wearer’s upper body during rehabilitation training with a lower limb rehabilitation exoskeleton. The instantaneous capture point is obtained by modeling the human-exoskeleton system and using the capture point theory. By comparing the stability region with instantaneous capture points of different gait phases, the balancing characteristics of different gait phases and changes to the equilibrium state in the gait process are analyzed. Based on a model of the human-exoskeleton system and the condition of balance of different phases, a trajectory correction strategy is proposed for the instability of the human-exoskeleton system caused by forward leaning of the wearer’s upper body. Finally, the reliability of the trajectory correction strategy is verified by carrying out experiments on the Zhejiang University Lower Extremity Exoskeleton. The proposed trajectory correction strategy can respond to forward leaning of the upper body in a timely manner. Additionally, in the process of the center of gravity transferred from a double-support phase to a single-support phase, the ratio of gait cycle to zero moment point transfer is reduced correspondingly, and the gait stability is improved.

Patients with stage II or III colorectal cancer (CRC) exhibit various clinical outcomes after radical treatments. The 5-year survival rate was between 50 and 87%. However, the underlying mechanisms of the variation remain unclear. Here we show that AMPKα1 is overexpressed in CRC patient specimens and the high expression is correlated with poor patient survival. We further reveal a previously unrecognized function of AMPKα1, which maintains high level of reduced glutathione to keep reduction–oxidation reaction (redox) homeostasis under stress conditions, thus promoting CRC cell survival under metabolic stress in vitro and enhancing tumorigenesis in vivo. Mechanistically, AMPKα1 regulate the glutathione reductase (GSR) phosphorylation possibly through residue Thr507 which enhances its activity. Suppression of AMPKα1 by using nano-sized polymeric vector induces a favorable therapeutic effect, especially when in combination with oxaliplatin. Our study uncovers a novel function of AMPKα1 in redox regulation and identifies a promising therapeutic strategy for treatment of CRC.

Background:Previous meta-analyses have suggested primary tumor location as a predictive factor for efficacy of anti–epidermal growth factor receptor (EGFR) therapies in patients with metastatic colorectal cancer (mCRC). However, the recent phase III TAILOR trial addressing this issue was not included in those analyses. This meta-analysis incorporated data from the TAILOR trial to evaluate the efficacy of chemotherapy plus anti-EGFR agents (cetuximab [Cet] or panitumumab [Pani]) versus chemotherapy alone forRASwild-type (wt) right- and left-sided mCRC.Patients and Methods:A PubMed-based literature search was conducted to identify randomized controlled trials (RCTs) studying the additional efficacy of Cet/Pani in combination with chemotherapy versus chemotherapy alone inRASwt left- and right-sided mCRC. Study-level pooled analyses of hazard ratios (HRs) for overall survival (OS) and progression-free survival (PFS) and odds ratios (ORs) for objective response rate (ORR) were performed.Results:Three first-line RCTs (CRYSTAL, PRIME, and TAILOR) and one second-line RCT (20050181) were included. Significant OS benefits from Cet/Pani were observed in the left-sided (HR, 0.76; 95% CI, 0.66–0.86) but not right-sided subgroups (HR, 0.99; 95% CI, 0.78–1.27). However, the addition of Cet/Pani to chemotherapy significantly improved PFS and ORR in both the left-sided (HR, 0.70; 95% CI, 0.57–0.86, and OR, 3.28; 95% CI, 1.95–5.51, respectively) and right-sided subgroups (HR, 0.76; 95% CI, 0.59–0.99, and OR, 1.78; 95% CI, 1.08–2.93, respectively).Conclusions:The addition of Cet/Pani to chemotherapy significantly benefits PFS and ORR in patients withRASwt right-sided mCRC, indicating that anti-EGFR therapies may remain an option for selected patients.

Background:The validity of progression-free survival (PFS) as a surrogate end point for overall survival (OS) in maintenance therapy trials of metastatic breast cancer (MBC) is uncertain. We aimed to compare treatment effect sizes and the strength of associations between OS and PFS in trials of maintenance therapy for MBC. Methods: We searched for randomized trials investigating maintenance chemotherapy, endocrine therapy or immunotherapy after first-line chemotherapy in MBC and selected those reporting results for both OS and PFS. Treatment effect size differences between OS and PFS by a ratio of hazard ratios (rHRs) with 95% confidence intervals [CIs] were evaluated using random effects analysis. Surrogacy were analyzed using a weighted linear regression model, correlations were evaluated by squared correlation R2. Results: We analyzed data from 16 trials and 3,898 patients that received maintenance chemotherapy, endocrine therapy or immunotherapy for MBC. In the all trial-level analysis, treatment effect sizes were 28% greater for PFS than for OS (combined rHR, 0.72; 95% CI, 0.62 to 0.85, P < 0.001), and the correlation coefficient R2 between PFS and OS was 18% (95% CI, 12% to 26%). Differences were greater with PFS than OS for trials of maintenance chemotherapy compared with observation (rHR, 0.72; 95% CI, 0.59 to 0.80, P < 0.001), and the correlation coefficient R2between treatment effects on PFS and on OS ranged from 12% (95% CI, 8% to 16%) when all trials were considered to 40% (95%CI, 30% to 54%) after exclusion of one highly influential trial by sensitivity analysis. Differences were also great for trials of maintenance endocrine therapy vs. observation (rHR, 0.54; 95% CI, 0.44 to 0.66), and immunotherapy vs. observation (rHR, 0.85; 95% CI, 0.80 to 0.91). Conclusion: PFS was greater than OS in the treatment effect sizes, which is a valid surrogate end point for OS to assess treatment effect in MBC maintenance therapy trials. PROSPERO registry: No. CRD42017071858; Support: ChiCTR-IIR-17014036, SYS-C-201801. Citation Format: Song E, Yao H, Yu Y, Ou Q, Wang Y. Progression-free survival is a surrogate of survival in maintenance therapy for metastatic breast cancer: Randomized trial level analysis [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr P1-16-07.

The genetic basis of colorectal cancer (CRC) and its clinical associations remain poorly understood due to limited samples or targeted genes in current studies. Here, we perform ultradeep whole-exome sequencing on 1015 patients with CRC as part of the ChangKang Project. We identify 46 high-confident significantly mutated genes, 8 of which mutate in 14.9% of patients: LYST, DAPK1, CR2, KIF16B, NPIPB15, SYTL2, ZNF91, and KIAA0586. With an unsupervised clustering algorithm, we propose a subtyping strategy that classisfies CRC patients into four genomic subtypes with distinct clinical characteristics, including hypermutated, chromosome instability with high risk, chromosome instability with low risk, and genome stability. Analysis of immunogenicity uncover the association of immunogenicity reduction with genomic subtypes and poor prognosis in CRC. Moreover, we find that mitochondrial DNA copy number is an independent factor for predicting the survival outcome of CRCs. Overall, our results provide CRC-related molecular features for clinical practice and a valuable resource for translational research.

Background Cholesterol is essential for colorectal cancer (CRC) cells. Cholesterol and its derivates are ligands for different nuclear receptors (NRs). However, which NR plays a key role in the downstream of cholesterol in CRC is unknown. This study aimed to elucidate the cholesterol-NRs axis in CRC. Methods We detected the levels of putative nuclear receptors of cholesterol metabolism in CRC cells treated with cholesterol deprivation. Then we built the RORα/γ knockdown and overexpression cell lines and examined the roles of RORα/γ in cell proliferation and migration of CRC in vitro and in vivo. Subsequently, we explored the downstream mechanisms and investigated the efficacy of the combination of Atorvastatin and RORα/γ agonists in mice. Results We found that the retinoic acid receptor-related orphan receptors RORα/γ levels increased most obviously in CRC cells after treated with cholesterol deprivation. RORα/γ can promote CRC cell proliferation and migration in vitro and in vivo. Mechanically, RORα/γ promotes c-myc degradation through activating ubiquitinase NEDD4 transcription. The combination of Atorvastatin and RORα/γ agonist SR1078 synergistically inhibited CRC cell growth and metastasis. Conclusions These findings demonstrate that cholesterol is essential for sustaining c-myc levels in CRC cells. The inhibition of RORα/γ by cholesterol and its derivatives is important for c-myc protein levels. And the combination of Atorvastatin and RORα/γ agonist might represent a therapeutic strategy in CRC.

BACKGROUND Revascularization is a successful therapeutic strategy for myocardial infarction. However, restoring coronary blood flow can lead to ischemia-reperfusion (I/R) injury. Low-dose 4-hydroxy-2-nonenal (HNE) therapy appears to play a key role in myocardial tolerance to I/R injury. We hypothesized that the positive effects of HNE on myocardial I/R injury may be UCP3-dependent. MATERIAL AND METHODS Adult male wild-type (WT) or UCP3 knockout (UCP3-/-) mice were pre-treated with the UCP inhibitor genipin or saline 1 h before ischemia and underwent 30-min coronary artery ligation followed by 24-h reperfusion. Mice were treated with intravenous HNE (4 mg/kg) or saline 5 min before reperfusion. Echocardiography was conducted to measure left ventricular end-diastolic posterior wall thickness (LVPWd), end-diastolic diameter (LVEDD), and fractional shortening (FS). Infarct size was measured by TTC staining. qRT-PCR and Western blotting were used to assess the expression of UCP3, UCP2, and the apoptosis markers cytochrome C and cleaved caspase-3. RESULTS HNE improved survival at 24 h post-MI in wild-type mice (p

Abstract Castration-resistant prostate cancer (CRPC), which is considered to contain cancer stem cells (CSCs), leads to a high relapse rate in patients with prostate cancer (PCa). However, the markers of prostate CSCs are controversial. Here we demonstrate that CD51, in part, correlates with the poor prognosis of PCa patients. Further, we find that CD51 is a functional molecule that is able to promote the malignancy of PCa through enhancing tumor initiation, metastatic potential, and chemoresistance. Moreover, we find that elevated CD51 expression in PCa specimens correlates with p53 loss of function. Mechanistically, we demonstrate that p53 acts via Sp1/3 to repress CD51 transcription, and CD51 is required for PCa stemness and metastasis properties, and is downregulated by p53. Taken together, these results indicate that CD51 is a novel functional marker for PCa, which may provide a therapeutic target for the efficiently restricting PCa progression.

Rationale: The antitumor activity of high-dose ascorbate has been re-evaluated recently, but the mechanism underlying cell-specific sensitivity to ascorbate has not yet been clarified. Methods: The effects of high-dose ascorbate on gastric cancer were assessed using cancer cell lines with high and low expression of GLUT1 via flow cytometry and colony formation assays in vitro and patient-derived xenografts in vivo. Results: In this study, we demonstrated that gastric cancer cells with high GLUT1 expression were more sensitive to ascorbate treatment than cells with low GLUT1 expression. GLUT1 knockdown significantly reversed the therapeutic effects of pharmacological ascorbate, while enforced expression of GLUT1 enhanced the sensitivity to ascorbate treatment. The efficacy of pharmacological ascorbate administration in mice bearing cell line-based and patient-derived xenografts was influenced by GLUT1 protein levels. Mechanistically, ascorbate depleted intracellular glutathione, generated oxidative stress and induced DNA damage. The combination of pharmacological ascorbate with genotoxic agents, including oxaliplatin and irinotecan, synergistically inhibited gastric tumor growth in mouse models. Conclusions: The current study showed that GLUT1 expression was inversely correlated with sensitivity of gastric cancer cells to pharmacological ascorbate and suggested that GLUT1 expression in gastric cancer may serve as a marker for sensitivity to pharmacological ascorbate.

Ginsenoside Rh2 (GRh2) and ginsenoside Rg3 (GRg3) are primary bioactive components in Panax ginseng. The present study aimed to investigate the underlying mechanisms of apoptotic cell-death induced by GRh2 and GRg3 in human leukemia Jurkat cells. The Cell Counting kit-8 assay was used to determine cell proliferation. Apoptosis was detected by nuclear morphologic observation by Hoechst 33342 staining and Annexin V-allophycocyanin and 7-amino-actinomycin D assay. mitoTEMPO, a mitochondrial reactive oxygen species (ROS) scavenger, was used to examine the effects of mitochondrial ROS on cell viability and mitochondrial membrane potential (MMP). Finally, the expression levels of numerous mitochondrial-associated apoptosis proteins were assessed by western blot analysis. These results demonstrated that GRh2 and GRg3 inhibited cell growth and induced apoptosis, and that GRh2 had greater cytotoxicity than GRg3. GRh2 induced generation of more mitochondrial ROS compared with GRg3 in Jurkat cells; however, this effect was ameliorated by subsequent treatment with mitoTEMPO. Furthermore, excess mitochondrial ROS induced by GRh2 was more potent than GRg3 in inhibiting cell proliferation and reducing MMP. In addition, expression levels of apoptosis-associated proteins were significantly increased in Jurkat cells treated with GRh2 than GRg3. In conclusion, these findings suggested that GRh2 and GRg3 induce mitochondrial-associated apoptosis by increasing mitochondrial ROS in human leukemia Jurkat cells. GRh2 may more effectively inhibit cell growth and accelerate apoptosis than GRg3. This study provides a potential novel strategy for the treatment of acute lymphoblastic leukemia.

Objectives Although the genomic landscape of small‐cell carcinoma of the oesophagus (SCCE) has been dissected, its transcriptome‐level aberration and immune microenvironment status are unknown. Methods Using ultra‐deep whole transcriptome sequencing, we analysed the expression profile of nine paired SCCE samples and compared the transcriptome with public transcriptomic data set of normal oesophageal mucosa and other cancer types. Based on the transcriptome data, the immune signatures were investigated. The genomic data of 55 SCCE samples were also applied for immune checkpoint blockade therapy (ICBT) biomarker evaluation including microsatellite instability (MSI) status, tumor mutation burden (TMB) and neoantigen burden (TNB). Also, we evaluated the CD8, CD68 and programmed death‐ligand 1 (PD‐L1) in 62 retrospective SCCE samples with IHC assay. Results Differential expression analysis revealed that the cell cycle, p53, and Wnt pathways are significantly deregulated in SCCE. Immune microenvironment analysis showed that high leucocyte infiltration and adaptive immune resistance did occur in certain individuals, while the majority showed a relatively suppressive immune status. Immune checkpoints such as CD276 and LAG‐3 were upregulated, and higher M2 macrophage infiltration in tumor tissues. Furthermore, normal tissues adjacent to the tumors of SCCE presented a more activated inflammatory status than tumor‐free healthy controls. These observations showed that ICBT might benefit SCCE patients. As the critical biomarker of ICBT, TMB of SCCE was 3.64 with the predictive objective response rate 13.2%, while the PD‐L1‐positive rate was 43%. Conclusions Our study systematically characterized the immune microenvironment in small‐cell carcinoma of the esophagus and provided evidence that several patients with SCCE may benefit from immune checkpoint blockade therapy., In this study, transcriptomic aberrance and immune microenvironment of small‐cell carcinoma of the esophagus (SCCE) was comprehensively characterized with transcriptome and immunohistochemistry analysis. Assessment of immune checkpoint blockade (ICB) treatment biomarkers provides a rationale for use of ICB treatment in patients with SCCE.

Background Immune checkpoint inhibitors (ICIs) have achieved impressive success in different cancer types, yet responses vary and predictive biomarkers are urgently needed. Growing evidence points to a link between DNA methylation and anti-tumor immunity, while clinical data on the association of genomic alterations in DNA methylation-related genes and ICI response are lacking. Methods Clinical cohorts with annotated response and survival data and matched mutational data from published studies were collected and consolidated. The predictive function of specific mutated genes was first tested in the discovery cohort and later validated in the validation cohort. The association between specific mutated genes and tumor immunogenicity and anti-tumor immunity was further investigated in the Cancer Genome Altas (TCGA) dataset. Results Among twenty-one key genes involving in the regulation of DNA methylation, TET1-mutant (TET1-MUT) was enriched in patients responding to ICI treatment in the discovery cohort (P 0.05 in both two non-ICI-treated cohorts). In TCGA dataset, TET1-MUT was strongly associated with higher tumor mutational burden and neoantigen load, and inflamed pattern of tumor-infiltrating T lymphocytes, immune signatures and immune-related gene expressions. Conclusions TET1-MUT was strongly associated with higher ORR, better DCB, longer PFS, and improved OS in patients receiving ICI treatment, suggesting that TET1-MUT is a novel predictive biomarker for immune checkpoint blockade across multiple cancer types.

Background Carnitine palmitoyltransferases (CPTs) are capable of transporting fatty acids (FAs) into mitochondria to synthesize cardiolipin and initiate β-oxidation. The aim of this study was to identify the inhibition of CPT-mediated FA catabolism on oxaliplatin sensitivity and its therapeutic potential in gastrointestinal cancers. Methods We investigated oxaliplatin-induced metabolic alterations, disease control rates (DCRs) using gastric cancer (GC) and colorectal cancer (CRC) cells as well as CRC tissues from patients treated with FOLFOX or XELOX regimens (n = 82). The effects of CPT inhibitor perhexiline on the growth of GC and CRC were tested in vitro and in vivo. Results CPT1B and CPT2 were significantly upregulated when GC and CRC cells were treated with oxaliplatin, and increased CPT1B and CPT2 expression correlated with a worse DCR in patients receiving FOLFOX or XELOX. Suppression of CPT2 or delivering perhexiline inhibited the progression of GC and CRC cells by inducing apoptosis, which was due to the loss of cardiolipin, suppression of β-oxidation, destruction of mitochondrial membrane integrity and reduction in ATP production. This process enhanced the antitumor effect of oxaliplatin in vitro and in vivo. Especially, perhexiline treatment significantly suppressed the progression of GC and CRC in patient-derived xenograft (PDX) models, which was closely associated with CPT2 expression. Conclusions Activated expression of FA catabolism is related to the chemoresistance of patients with GC and CRC. Inhibition of CPT cooperates with oxaliplatin to suppress their growth, as CPT1B and CPT2 are potential biomarkers for platinum-based chemotherapy in GC and CRC.

Background Long non-coding RNAs (lncRNAs) play an unneglectable role in epigenetic regulation of cancer cells, including binding to proteins and regulating cellular metabolism. The aim of this study was to identify a certain lncRNA promoting the progress of advanced colorectal cancer (CRC) with a therapeutic perspective. Methods We screened out highly expressed lncRNAs using samples from patients with stage IV CRC compared with matched adjacent normal tissues. The proteins interacted with linc00920 was confirmed with RNA pull-down and RNA immunoprecipitation (RIP) assay. The proliferation and metabolic alteration of CRC under linc00920 inhibition were tested in vitro and in vivo. Results Linc00920 was upregulated in CRC with poor overall survival and inhibition of linc00920 resulted in impaired growth of CRC cell lines. Moreover, knocking down of linc00920 was consistent with a lower level of insulin-like growth factor 2 mRNA-binding protein 2 (IGF2BP2), which is known as a m6A ‘reader’ and mRNA stabilizer. Linc00920 bound to the ubiquitination sites of IGF2BP2 and prevented its autophagic degradation, maintaining the MYC-mediated glycolysis in CRC. Moreover, inhibition of linc00920 suppressed the proliferation of tumors from patient-derived xenograft (PDX) models. Conclusions Linc00920-IGF2BP2-MYC axis promotes the progress of CRC as a promising therapeutic target.

Background: Accumulating evidence has demonstrated that microRNAs regulate diverse tumorigenic processes and play important roles in tumor metastasis and growth. Recently, miR-125b-2-3p was identified as a meaningful prognostic factor for predicting chemotherapeutic sensitivity in advanced colorectal cancer (CRC). However, the biological functions and molecular mechanisms of miR-125b-2-3p in the chemotherapy of advanced CRC have yet to be elucidated. Methods: MiR-125b-2-3p expression was detected by real-time PCR in CRC tissues. Gain-of-function experiments were performed to assess the effect of miR-125b-2-3p on CRC growth, metastasis, invasion and drug sensitivity in vitro and in vivo. Based on multiple databases, the competitive endogenous RNAs (ceRNAs) and target genes for miR-125b-2-3p were predicted and have been confirmed by bioinformatic analysis, luciferase reporter assays, rescue experiments and western blot assays. Findings: MiR-125b-2-3p expression was significantly downregulated in CRC tissues and cell lines. MiR-125b-2-3p overexpression was correlated with lower proliferation rates, fewer metastases and better chemotherapeutic sensitivity in vitro and in vivo. Mechanistically, miR-125b-2-3p was regulated by lncRNA XIST and influenced the expression of the WEE1 G2 checkpoint kinase (WEE1). Interpretation: These findings suggested that the lncRNA XIST-miR-125b-2-3p-WEE1 axis not only regulated CRC growth and metastasis but also contributed to chemotherapeutic resistance, thus identifying a potential therapeutic target for CRC. Funding: National Natural Science Foundation of China (81572392), Guangdong special support program (2016TQ03R614), National Key RD Natural Science Foundation of Guangdong Province (2014A030312015); Science and Technology Program of Guangdong (2019B020227002); Science and Technology Program of Guangzhou (201904020046, 201803040019, 201704020228). Declaration of Interest: The authors declare no conflicts of interest. Ethical Approval: This study was approved by the institutional ethics review board of SYSUCC (Guangzhou, China). Our animal study was approved by the Institutional Animal Care and Use Committee of Sun Yat-sen University.

Acute lymphoblastic leukaemia (ALL) is the most prevalent childhood malignancy. Although most children with ALL are cured, there is still a group of patients for which therapy fails owing to severe toxicities and drug resistance. Ginsenoside Rh2 (GRh2), a major bioactive component isolated from Panax ginseng, has been shown to have a therapeutic effect on some tumors. However, the molecular mechanisms of cell death induced by 20(S)-GRh2 in ALL cells remains unclear. In this study, we showed that 20(S)-GRh2 inhibited the cell growth and induced mitochondria-dependent apoptosis and autophagy. But it has no cytotoxic effect on human normal blood cells. Furthermore, autophagy plays a protective role in 20(S)-GRh2-induced apoptosis in ALL cell lines and human primary ALL cells. We demonstrated that either genetic or pharmacologic inhibition of autophagy could be more effective in reducing viability and enhancing 20(S)-GRh2-induced toxicity than 20(S)-GRh2 treatment alone. In addition, inhibition of autophagy could aggravate mitochondrial ROS generation and mitochondrial damage, and then accelerate mitochondria-dependent apoptosis. Taken together, these results suggest that inhibition of autophagy can sensitize ALL cells towards 20(S)-GRh2. The appropriate inhibition of autophagy could provide a powerful strategy to increase the potency of 20(S)-GRh2 as a novel anticancer agent for ALL therapy.

Background Long non-coding RNAs (lncRNAs) are involved in innate and adaptive immunities in cancers by mediating the functional states of immunologic cells, pathways, and genes. However, whether lncRNAs could serve as effective biomarkers for molecular classification and prediction of cancer immunotherapy efficacy are largely unknown. Methods This study analyzed lncRNA and genomic data of 348 atezolizumab-treated bladder cancer patients from phase II IMvigor 210 trial and 3,021 patients in lung cancer, breast cancer, bladder cancer, and melanoma cohorts from The Cancer Genome Atlas. We investigated lncRNA-based immune subtypes associated with cancer immunotherapy efficacy. We built a novel lncRNA score using computational algorithms and integrated it with programmed-death ligand 1 (PD-L1) expression and tumor mutation burden (TMB) to achieve accurate immunotherapeutic prediction. Results The results from IMvigor 210 trial showed that four distinct microenvironment-based subtypes characterized by lncRNA expression and tumor specific cytotoxic T lymphocytes (CTLs) infiltration had significant difference in overall survival (OS) (hazard ratio [HR] 0.77, 95%CI 0.67–0.88; P = 0.0002), with the greatest benefits in Immune-Active Class, followed by Immune-Exclusion Class, Immune-Dysfunctional Class, and Immune-Desert Class. High NKILA lncRNA expression was identified as a negative predictor of immunotherapeutic OS benefits and a critical regulator of dysfunctional immune response. Patients with low- versus high- lncRNA scores were associated with significantly longer OS in IMvigor 210 trial (HR 0.32, 95% CI 0.24–0.42; P Conclusion We suggested immunotherapy for patients in Immune-Functional Class, especially for those in high CTL Immune-Active Class. Multiomics comprising lncRNA score, PD-L1 expression and TMB could accurately predict immunotherapy efficacy. Legal entity responsible for the study Herui Yao. Funding Grants from the National Natural Science Foundation of China (81372819, 81572596, U1601223), the Natural Science Foundation of Guangdong Province (2017A030313828), the Guangzhou Science and Technology Program (201704020131), the Sun Yat-Sen University Clinical Research 5010 Program (2018007). Disclosure All authors have declared no conflicts of interest.

Background Tumour cells often exhibit altered energy metabolism for tumour progression, but whether long noncoding RNAs (lncRNAs) are involved in this process remains elusive. We aimed to identify novel lncRNAs significantly affecting the development of Esophageal Squamous Cell Carcinoma (ESCC) and investigate the metabolism-related mechanisms. Methods A siRNA library was established using top 100 lncRNAs highly expressed in ESCC screened from TCGA database and transfected into ESCC cells to identify lncRNAs that significantly affected cell glucose metabolism and proliferation. RNAscope in situ hybridization, qRT-PCR and RNAi assays were performed to investigate the functional role of lncRNA AGPG (Actin Gamma 1 Pseudogene) and clinical relevance. In vivo, cell-based and patient-derived xenograft (PDX) models were used to further explore roles of AGPG in ESCC tumorigenesis. RNA pull-down, mass spectrometry analyses, western blot and RNA-binding protein immunoprecipitation (RIP) were performed to identify interaction proteins and related mechanisms. Results We identified that lncRNA AGPG was markedly increased in ESCC and correlated with poor prognosis. AGPG depletion significantly repressed ESCC cell proliferation and glycolytic activity including decreased glucose uptake and increased lactate production. In vivo experiments further showed that silencing AGPG decreased tumours growth in cell-based xenografts and PDX models. Mechanistically, AGPG could interact with 6-phosphofructo-2-kinase/fructose-2, 6-biphosphatase 3 (PFKFB3) in the nucleus, thus enhancing PFKFB3 stability via preventing its ubiquitination and proteosomal degradation. Subsequently, AGPG-PFKFB3 promoted glycolysis and dysregulated cell cycle by suppressing p27 and over-activating cyclin-dependent kinase 1(CDK1), thereby leading to G1/S cell cycle progression. Additionally, TP53 could bind to the promoter of AGPG to negatively regulate its expression. Conclusions We first identified that AGPG, which is regulated by TP53, is playing a pivotal role in glucose metabolism remodelling and cell proliferation through enhancing PFKFB3 stability, thus facilitating the development of ESCC. Collectively, our study suggests that TP53-AGPG-PFKFB3 axis might serve as potential biomarkers and therapeutic targets in ESCC.

Mesenchymal stem cells (MSCs) have recently been shown to home to tumors and contribute to the formation of the tumor-associated stroma. In addition, MSCs can secrete paracrine factors to facilitate tumor progression. However, the involvement of MSC-derived cytokines in colorectal cancer (CRC) angiogenesis and growth has not been clearly addressed. In this study, we report that interleukin-8 (IL-8) was the most highly upregulated pro-angiogenic factor in MSCs co-cultured with CRC cells and was expressed at substantially higher levels in MSCs than CRC cells. To evaluate the effect of MSC-derived IL-8 on CRC angiogenesis and growth, we used MSCs that expressed small hairpin (interfering) RNAs (shRNA) targeting IL-8 (shIL-8-MSCs). We found that MSC-secreted IL-8 promoted human umbilical vein endothelial cell (HUVEC) proliferation and migration, tube-formation ability and CRC cell proliferation. Additionally, in vivo studies showed that MSCs promoted tumor angiogenesis partially through IL-8. Taken together, these findings suggest that IL-8 secreted by MSCs promotes CRC angiogenesis and growth and can therefore serve as a potential novel therapeutic target.

Outdoor SVG is the SVG that is derived from conventional SVG, and can be used on sites of limited space. After repeated technical upgrading, this kind of SVG is functioning more and more stably, and is well received by clients. However, since the internal space of outdoor SVG is greatly compressed, and overheat trip still occurs in “extreme environment”. Because the “extreme environment” is just occasional, increase in number or power of fans will greatly increase equipment cost and failure points, most SVG manufacturers are not willing to adopt this strategy. In this paper, the outdoor SVG “overheat capacity reduction” control strategy based on wireless bluetooth humidity and temperature monitoring system is proposed, which ensures that the equipment can run safely in “extreme environment”, saving labor and materials remarkably.

Anoikis is a critical obstacle to cancer metastasis. Colorectal cancer (CRC) exhibits a high rate of metastasis, leading to death, and the mechanisms involved in anoikis resistance are still unclear. We identified that the fatty acid oxidation (FAO) pathway was activated in detached CRC cells. Multiple genes in the FAO pathway, specifically the rate-limiting enzyme CPT1A, were upregulated in CRC cells grown in suspension. Reactive oxygen species elimination mediated by CPT1A in CRC cells was vital to anoikis resistance. In vivo experiments showed that CPT1A-suppressed CRC cells colonized the lung at a much lower rate than normal CRC cells, suggesting that CPT1A-mediated FAO activation increased metastatic capacity. In clinical tissue specimens from CRC patients, elevated expression of CPT1A was observed in metastatic sites compared with primary sites. Our results demonstrate that CPT1A-mediated FAO activation induces CRC cells to resist anoikis, suggesting that CPT1A is an attractive target for treating metastatic CRC.

In order to solve the problem of low voltage at a terminal rural power grid 66kV substation, a 5000-kilovar SVG is arranged in the substation to elevate and stabilize the 10kV bus voltage. Since put into operation, SVG runs well, solving the problem of low voltage while lowering the loss of the entire line. However, in some cases, people find that while stabilizing the bus voltage, SVG causes voltage drop in adjacent substation, and act of the intelligent capacitor unit in the adjacent substation, thus leading to opposite reactive response of SVG, affecting voltage stabilization of the two substations and adding up to equipment loss. This paper analyzes this phenomenon and has proposed an approach of correction.

We use Thevenin battery model and Kalman filter algorithm to online estimate lithium-ion battery pack state of charge (SOC) in this paper. In order to improve the accuracy of the model we use least square method and Dual Kalman filter (DEKF) algorithms to identify the parameters of model. The battery model can reflect the true state of internal battery well. The principle of Kalman filter algorithm is introduced. The relevant battery testing laboratory is designed. The algorithm has better accuracy when online estimate SOC and adapt to the environment well from experimental results. Finally, the convergence and robustness of DEKF algorithm are verified. It solves the problems that initial estimates are not accuracy and cumulative error.

A temperature prediction model based on Self-adaption Particle Swarm Optimization (SAPSO) and Extreme Learning Machine (ELM) is proposed in this paper. The nano-iron powder decomposing furnace temperature prediction model is established based on ELM. ELM, a neural network, is developed rapidly in recent years, but it requires a lot of hidden layer neurons to achieve ideal prediction accuracy. In order to solve this problem, SAPSO is used to optimize ELM input weight matrix and the hidden layer threshold value in this paper, which simplifies the ELM. The accuracy of this method is verified by choosing the related factors, using field data and simulation. Different numbers of neurons are set to compare with ordinary ELM. And the test shows that SAPSO-ELM temperature prediction model has better generalization.

Background: Tumor mutational burden (TMB) has been widely studied as a predictive biomarker of response to immune checkpoint inhibitors (ICIs). Besides, evidence suggests frameshift indels are a highly immunogenic mutational class and thus a potentially superior biomarker. However, the general prognostic impact of TMB and indel burden in patients with solid tumors has not been systematically investigated. Methods: We analyzed 20 primary solid cancer types from The Cancer Genome Atlas (TCGA) database. Clinicopathologic factors, TMB and indel burden were collected or calculated. For each cancer type, the impact of TMB or indel burden on overall survival (OS) was evaluated using the Kaplan-Meier method and Cox regression with the method of inverse probability of treatment weighting. Results: Twenty cancer types from 6,035 patients were analyzed. Survival analysis showed that TMB had a significant impact on OS in 14 out of these 20 cancer types. According to the general survival impact of TMB, they could be classified into three groups, namely the TMB-Worse (eight cancer types), TMB-Better (six cancer types) and TMB-Similar (six cancer types) group, in which higher TMB was associated with inferior, superior, or similar OS, respectively. The survival impacts of TMB in the TMB-Worse and TMB-Better groups were generally consistent when limited to genes from two FDA-approved panels. Notably, in two out of the six cancer types in the TMB-Similar group, the indel burden significantly affected OS. Conclusions: TMB, as well as indel burden, has divergent prognostic impact in different cancer types, thus could be incorporated in prognostication and risk stratification. More importantly, the general prognostic impact should be taken into account when establishing the predictive function of TMB to ICI treatment.

This cohort study analyzes the medical records of 47 721 patients with various cancer types with POLE/POLD1 mutations to evaluate whether the mutations were associated with immunotherapy outcomes.

Background/Aims: Small cell carcinoma of the esophagus (SCCE) is a rare and highly deadly malignancy with rapid disease progress and extensive metastasis, whereas present therapy for the cancer is limited, making the identification of a new treatment strategy an area of intense research. We previous reported the genomic characteristics and alterations of the SCCE, but the transcriptome abnormality of the cancer has not been investigated yet. Method: we performed an RNA Sequencing on nine paired samples from patients with SCCE and conducted a series of comprehensive analysis on the data. DESeq2 and GSEA were applied on transcriptome of SCCE tumor tissue and their para-normal to identify differential expressive genes and associated pathways. Besides, with MCP-counter and a list of immunity related genes, we also compared immune signal of the expression data in pair. By applying a rigorous batch removing strategy, including preprocessing raw data in identical method and RUVSeq, transcriptome data of healthy esophageal mucosa from GTEx was involved as control samples in comparison. In addition, we applied CIBERSORT to calculate the fraction of different tumor infiltrated leukocytes (TILs) of SCCE and compare the number with those of other cancers that have been proven to be potentially responsive to immunotherapy. Results: We identified 1486 and 1782 genes were significantly up-regulated and down-regulated in SCCE tumor tissue against their para-normal, respectively. Pathway analysis revealed the E2F Target and G2M checkpoint are significantly enriched in tumor tissue. Immune signal analysis across the samples in pair showed that in 3 out of 9 paired sample, immunity reaction upregulates in tumor tissue. T-SNE cluster across transcriptome data of SCCE tumor tissue, their para-normal and healthy esophageal mucosa showed the para-normal tissue, namely the normal tissue adjacent to tumor, is more similar to tumor tissue than the healthy. CIBERSORT analysis showed the difference in TILs fraction between SCCE and the compared cancers. For example, the relative fraction of plasma cell of SCCE is higher than those of ESCC(P=0.040), CIN type of STAD(P=0.040), while lower than those of SCLC(P=0.020). M2 macrophages is higher in SCCE than ESCC(P=0.002), EAC(P Conclusion: In summary, our analysis provides transcriptome evidence that immunotherapy might be an efficient treatment strategy for small cell esophagus cancer. Citation Format: Qi Zhao, Yan-xing Chen, Ying-nan Wang, Qi-nian Wu, Hui Sheng, JIa-jia Hu, Yun-xin Lu, Ze-xian Liu, Zhi-xiang Zuo, Feng Wang, Rui-hua Xu. Systematic transcriptome analysis of small cell carcinoma of esophagus suggests a possibility for immunotherapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 4242.

Background/Aims: Colorectal cancer (CRC) is a common cancer worldwide with increasing morbidity in China. Approximately 5% CRC are caused by well-defined hereditary CRC syndromes, whose inherited susceptibility account for nearly one-third of CRC predisposition. Most of hereditary CRC syndromes were diagnosed at the age of lower than 70, whereas little is known about the mutational profiles Chinese patients and their correlations with clinical features. Method: In our present research, we retrospectively collected 731 patients who were diagnosed as CRC from Sun Yat-sen University Cancer Center, with ages lower than 70. All of them received genetic mutation detection consisting of a set of 14 specific inherited genes, which used target area capture and the second-generation high throughput sequencing technology. The characteristics of he patients with or without mutations were compared. We applied several comparison analysis to investigate the germline mutations distribution of genes that associated with the defined hereditary CRC syndromes in Chinese cohort. Results: 397 patients (54.3%) harbored functionally variants of the 14 genes in CRC patients that diagnosis lower than 70. Intriguingly, we observed no statistical difference for clinical factors such as age, sex, smoke, drink, family history, pathology grade, clinic stage and overall survival between groups. In Mut group, the mutation frequency of each genes are MLH1(21.4%), MSH2(17.6%), MSH6(15.1%), PMS2(9.3%), EPCAM(8.3%), MLH3(8.1%), AXIN2(7.1%), PMS1(4.0%), MUTYH(12.8%), APC(12.3%), STK11(3.5%), BMPR1A(1.8%), SMAD4(0.3%), PTEN(1.0%). Genome mutual exclusivity analysis has identified a set of genes such as MLH1, MSH2, MUTYH, MSH6, APC and PMS2, for which most pairwise combinations are found rarely cooccurred. In correlation analysis between mutational genes and clinical features, pathology grade correlates strongly with PMS1 and SMAD4(P Conclusion: The clinic features of two groups have no statistic difference, which suggested other genes undiscovered may play a partial role in pathogenesis of patients of non-MUT group. In summary, we conducted a retrospective study to lead a comprehensive comparison of mutational profiles between MUT and non-MUT hereditary CRC patients, which supplemented the limited data of hereditary CRC in China. Citation Format: Feng Wang, Qi Zhao, Teng-jia Jiang, Ying-nan Wang, Fu-rong Liu, Jia-jia Hu, Pei-rong Ding, Zhi-zhong Pan, Jian-yong Shao, Rui-hua Xu. Germline mutational profileof Chinese patients with colorectal cancer and diagnosed lower than 70 years [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 4161.

This systematic review and meta-analysis examines randomized clinical trials (RCTs) of immune checkpoint inhibitors to evaluate the use of milestone rate and milestone restricted mean survival time as intermediate end points in immune checkpoint inhibitor trials., Key Points Question What intermediate end points could be useful in randomized clinical trials studying immune checkpoint inhibitors? Findings In this systematic review and meta-analysis of 26 trials studying immune checkpoint inhibitors in 12 892 participants, the ratio of milestone restricted mean survival time for overall survival was more strongly correlated with the overall survival hazard ratio than the ratio of overall survival milestone rates. Meaning Milestone restricted mean survival time could be studied as a potential intermediate end point for overall survival in future trials of immune checkpoint inhibitors., Importance Immune checkpoint inhibitors (ICIs) have unique patterns of response and survival that differ from conventional chemotherapies. Novel intermediate end points are urgently required to detect the early signals of ICI activity. Objective To evaluate milestone rate (Kaplan-Meier estimates of survival probabilities at given time points) and milestone restricted mean survival time (RMST, the area under survival curves up to given time points) as potential intermediate end points for ICI trials. Data Sources Electronic databases (pre-MEDLINE, MEDLINE, Embase, and the Cochrane Central Register of Controlled Trials) were searched for randomized clinical trials published between January 1, 2000, and December 31, 2017. Study Selection Phase 2 and phase 3 randomized clinical trials evaluating ICIs in advanced solid tumors. Data Extraction and Synthesis Two investigators extracted the data and reconstructed individual patient data to estimate the milestone rate or milestone RMST from the published Kaplan-Meier curves. Main Outcomes and Measures Trial-level milestone rates or milestone RMSTs were estimated for 6-month and 9-month progression-free survival (PFS) and 9-month and 12-month overall survival (OS). A weighted linear regression model evaluated the correlations of ratios of milestone rates or milestone RMSTs with OS hazard ratios (HRs). Results Twenty-six trials examining 8 different tumor types were identified, including 12 892 patients. Overall survival HR was correlated with the ratio of 9-month OS milestone rate (R2 = 0.45; 95% CI, 0.27-0.74), and with the ratio of 12-month OS milestone rate (R2 = 0.40; 95% CI, 0.22-0.70). The ratio of 9-month OS milestone RMST (R2 = 0.60; 95% CI, 0.28-0.74) and ratio of 12-month OS milestone RMST were correlated with OS HR (R2 = 0.64; 95% CI, 0.42-0.78). No correlations were observed between OS HR and the ratio of 6-month or 9-month PFS milestone rates or milestone RMSTs. Conclusions and Relevance Ratios of OS milestone RMSTs had a stronger correlation with OS HRs than ratios of OS milestone rates, whereas ratios of PFS milestone rates and ratios of PFS milestone RMSTs were not correlated with OS HRs. The OS milestone RMST could be further studied as an intermediate end point in future ICI trials.