Efficacy of Endoscopic Tissue Adhesive in Patients with Gastrointestinal Tumor Bleeding.

BACKGROUND: Gastrointestinal tumors bleeding remains a significantly clinical challenge due to its resistance to conventional endoscopic hemostasis methods. While the efficacy of endoscopic tissue adhesives (ETA) in variceal bleeding has been established, its role in gastrointestinal tumor bleeding (GITB) remains ambiguous. AIMS: This study aims to assess the feasibility and effectiveness of ETA in the treatment of GITB. METHODS: The study enrolled 30 patients with GITB who underwent hemostasis through Histoacryl® tissue glue injection. Hemostasis success rates, ETA-related adverse events, and re-bleeding rates were evaluated. RESULTS: ETA application achieved successful hemostasis at all tumor bleeding sites, with immediate hemostasis observed in all 30 (100.0%) patients. Among the initially hemostasis cases, 5 patients (17.0%) experienced re-bleeding within 30 days, and the 60 day re-bleeding rate was 20.0% (6/30). Expect for one case of vascular embolism, no adverse events related with ETA application were reported. The 6 month survival was 93%. CONCLUSION: ETA demonstrated excellent immediate hemostasis success rate in GITB cases and showed promising outcomes in prevention re-bleeding.

Identification and Validation of the Glycolysis and Immune-related Gene Signature for Prognosis in Colorectal Cancer.

BACKGROUND/AIM: Glycolysis has a role in regulating the tumor immune microenvironment. However, the functions and clinical role for facilitating the prognosis prediction of colorectal cancer (CRC) based on glycolysis and immune-related genes remain to be identified. MATERIALS AND METHODS: Genes associated with glycolysis and immunity (GI) were identified from established databases (MSigDB and ImmPort). The TCGA (training cohort) and GSE39582 (validation cohort) datasets were used. Cox regression and least absolute shrinkage and selection operator (LASSO) Cox regression analyses were applied for model construction. The prognostic power of the GI signature was examined by multivariate Cox regression analysis. The correlations between the GI signature, immune cell infiltration and immune checkpoint blockade (ICB) genes were analyzed. To further validate the identified gene signature, quantitative RT-PCR was performed. Cell proliferation assays were conducted for CCK8 detection. RESULTS: A new GI model was constructed, and this signature may serve as an independent prognostic biomarker in CRC. The GI signature remained an effective tool for predicting prognosis among each clinical subgroup. This signature was related to immune cell infiltration of myeloid dendritic cells, cancer-associated fibroblasts (CAFs), CD4+ and CD8+ T cells and response to the ICB immunotherapy-related genes IDO1, BTLA, PD-L1 and PD-L2. In addition, our findings showed that PMM2, IL20RB, and NTF4 exhibited high expression levels in CRC. The upregulation of these genes resulted in the promotion of the proliferation of CRC cells. CONCLUSION: This novel prognostic signature contributed to CRC risk stratification and survival prediction based on glycolysis and immune status.

A Study of the Detection of SARS-CoV-2 by the Use of Electrochemiluminescent Biosensor Based on Asymmetric Polymerase Chain Reaction Amplification Strategy.

A new and reliable method has been constructed for detecting severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2) open reading frames 1ab (ORF1ab) gene via highly sensitive electrochemiluminescence (ECL) biosensor technology based on highly efficient asymmetric polymerase chain reaction (asymmetric PCR) amplification strategy. This method uses magnetic particles coupled with biotin-labeled one complementary nucleic acid sequence of the SARS-CoV-2 ORF1ab gene as the magnetic capture probes, and [Formula: see text]-labeled amino-modified another complementary nucleic acid sequence as the luminescent probes, and then a detection model of magnetic capture probes-asymmetric PCR amplification nucleic acid products-[Formula: see text]-labeled luminescent probes is formed, which combines the advantages of highly efficient asymmetric PCR amplification strategy and highly sensitive ECL biosensor technology, enhancing the method sensitivity of detecting the SARS-CoV-2 ORF1ab gene. The method enables the rapid and sensitive detection of the ORF1ab gene and has a linear range of 1-[Formula: see text] copies/[Formula: see text], a regression equation of [Formula: see text] = [Formula: see text] + 2919.301 ([Formula: see text] = 0.9983, [Formula: see text] = 7), and a limit of detection (LOD) of 1 copy/[Formula: see text]. In summary, it can meet the analytical requirements for simulated saliva and urine samples and has the benefits of easy operation, reasonable reproducibility, high sensitivity, and anti-interference abilities, which can provide a reference for developing efficient field detection methods for SARS-CoV-2.

A Study of the Detection of SARS-CoV-2 ORF1ab Gene by the Use of Electrochemiluminescent Biosensor Based on Dual-Probe Hybridization.

To satisfy the need to develop highly sensitive methods for detecting the severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2) and further enhance detection efficiency and capability, a new method was created for detecting SARS-CoV-2 of the open reading frames 1ab (ORF1ab) target gene by a electrochemiluminescence (ECL) biosensor based on dual-probe hybridization through the use of a detection model of "magnetic capture probes-targeted nucleic acids-Ru(bpy)32+ labeled signal probes". The detection model used magnetic particles coupled with a biotin-labeled complementary nucleic acid sequence of the SARS-CoV-2 ORF1ab target gene as the magnetic capture probes and Ru(bpy)32+ labeled amino modified another complementary nucleic acid sequence as the signal probes, which combined the advantages of the highly specific dual-probe hybridization and highly sensitive ECL biosensor technology. In the range of 0.1 fM~10 µM, the method made possible rapid and sensitive detection of the ORF1ab gene of the SARS-CoV-2 within 30 min, and the limit of detection (LOD) was 0.1 fM. The method can also meet the analytical requirements for simulated samples such as saliva and urine with the definite advantages of a simple operation without nucleic acid amplification, high sensitivity, reasonable reproducibility, and anti-interference solid abilities, expounding a new way for efficient and sensitive detection of SARS-CoV-2.

Subgroup Identification with Gene Expression Profiles of Adipose Tissue in Patients with Coronary Artery Disease.

Among many diseases, coronary artery disease (CAD) is the primary cause of mortality and morbidity worldwide. With the aim of revealing the underlying genetic characteristics of the CAD subtypes, we recruited patients with CAD and categorized them into subgroups according to the transcriptome expression profiles of the adipose tissue.With the removal of the batch effect, consensus clustering was employed to determine the subgroup numbers. Subgroup-specific genes were determined to conduct analysis of Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG). Weighted gene co-expression network analysis (WGCNA) revealed the subgroup-specific WGCNA modules. Moreover, gene set enrichment analysis (GSEA) was conducted. Overrepresentation enrichment analysis (OEA) of subgroup-specific signatures was also conducted to reveal the significant gene module associated with the corresponding clinical characteristics.After the removal of the batch effect, 77 CAD objects were divided into three subgroups. It was observed that the patients in subgroup III tended to be fat. After analyzing the dominant pathways of each subgroup, we discovered that the protein digestion and absorption pathway was specifically upregulated in subgroup I, which might result from the lowest proportion of the epicardial adipose tissue (EAT) sample. Moreover, subgroup II patients had genetic characteristics of high expression of complement and coagulation cascades and TNF signaling pathway. Furthermore, Th17 cell differentiation was significantly upregulated in subgroup III, indicating that Th17 cell differentiation is related to the clinical characteristics of body mass index (BMI).In conclusion, the genetic classification of CAD subjects indicated that subjects from different subgroups may exhibit specific gene expression patterns, suggesting that more personalized treatment should be applied to patients in each subgroup.

Effectiveness and Safety of Cryoablation in Patients With Atrial Fibrillation Episodes of <24 h Duration: A Propensity-Matched Analysis.

Background: Paroxysmal atrial fibrillation (AF) is closely related to pathophysiologic processes and clinical outcomes. However, it is uncertain whether cryoablation of pulmonary veins isolation is effective and safe for patients with symptomatic and drug refractory AF episodes of <24-h duration. Methods: The patients were designed into Group A (253 patients with paroxysmal AF episodes of <24-h duration) and Group B (253 patients with paroxysmal AF lasting for 24 h or longer) on a 1:1 basis by identical propensity scores. Mortality, stroke/transient ischemic attack (TIA), and complications relevant to the cryoablation procedure were compared, and recurrence of atrial tachyarrhythmia was analyzed for clinical independent predictors. Results: The rate of atrial tachyarrhythmia recurrence was 21.74% in Group A and 30.04% in Group B, respectively (P = 0.042). At 12-month follow-up from the procedure, lower incidences of stroke/TIA endpoint of the patients were observed in Group A compared with Group B by Kaplan-Meier analysis [HR 0.34 (0.13-0.87), P = 0.025]. No significant differences in mortality and complications relevant to the cryoablation procedure were observed between Group A and Group B. Moreover, adjusted multivariable Cox regression analysis showed that <24-h paroxysmal AF type (HR 0.644, 95% CI: 0.455-0.913, P = 0.014) and left atrium diameter (LAD) (>40 mm) (HR 1.696, 95% CI: 1.046-2.750, P = 0.032) were independently associated with the incidence of recurrence of atrial tachyarrhythmia in the study. Conclusion: Our findings indicated that <24-h paroxysmal AF type was obviously associated with an increased success rate of cryoablation and reduced incidence of stroke/TIA during the follow-up period. Therefore, there is superior effectiveness and similar safety in patients with AF episodes of <24-h duration compared with patients with longer paroxysmal AF duration.

Recurrent atrial fibrillation after initial cryoballoon ablation: New perspectives for intensive ablation in right superior pulmonary vein ostium for atrial fibrillation.

A significant proportion of patients with recurrent atrial fibrillation (AF) require repeat radiofrequency (RF) ablation after cryoballoon (CB) ablation. However, little is known about the pulmonary vein (PV) potential reconnection to left atrium and localization of gaps in the initial lesion sets following cryoablation in patients with recurrent AF. The data of 29 consecutive patients with repeat RF ablation for recurrent AF were analyzed. During the second ablation procedures, PV foci of AF were explored in 116 PVs by the CARTO system. All patients had complete PV isolation from initial cryoablation procedure. The fluoroscopy duration, mean cryoablation time and mean cryoablation frequency were lowest for the right superior pulmonary vein (RSPV) (58.69 ± 9.18 s, 185.10 ± 49.25 s and 1.07 ± 0.26; p = 0.024, p = 0.042 and p = 0.032). A significantly higher incidence of conduction gaps per patient was found for the RSPVs compared to the other PVs (p < 0.05 or p < 0.01). For RSPVs, it seemed that gaps were predominantly located at the anterior segment (22 gaps) and inferior segment (22 gaps). RSPV reconnection was independently related to a lower risk of major adverse events after the second ablation during follow up in the study patients (HR 0.275, 95%CI 0.078-0.967, p = 0.044). AF recurrence in patients after cryoablation is significantly associated with conduction gaps in the anterior and inferior segments of RSPVs. Various ablation strategies of close touch of CB on anterior and inferior segments of RSPV ostium, more freezing time and frequency for RSPV may help achieving durable PV isolation during follow up.

Long non-coding RNA PROX1-AS1 knockdown upregulates microRNA-519d-3p to promote chemosensitivity of retinoblastoma cells via targeting SOX2.

OBJECTIVE: Long non-coding RNAs (lncRNAs) and microRNAs (miRNAs) participate in tumor progression, while the role of PROX1-antisense RNA1 (PROX1-AS1) sponging miR-519d-3p in retinoblastoma (RB) remains largely unknown. We aim to explore the effect of the PROX1-AS1/miR-519d-3p/sex determining region Y-box 2 (SOX2) in chemosensitivity of RB cells. METHODS: Expression of PROX1-AS1, miR-519d-3p and SOX2 in RB tissues and cells was determined. The drug-resistant cell lines were established and respectively intervened with PROX1-AS1 or miR-519d-3p expression to explore their roles in drug resistance and malignant behaviors of the drug-resistant cells. The binding relationships between PROX1-AS1 and miR-519d-3p, and between miR-519d-3p and SOX2 were evaluated. RESULTS: PROX1-AS1 and SOX2 were upregulated while miR-519d-3p was downregulated in RB tissues and cells, especially in drug-resistant cells. The PROX1-AS1 inhibition or miR-519d-3p elevation suppressed the drug resistance, proliferation, migration and invasion, and promoted apoptosis of the drug-resistant RB cells. Moreover, PROX1-AS1 sponged miR-519d-3p and miR-519d-3p targeted SOX2. CONCLUSION: PROX1-AS1 knockdown upregulates miR-519d-3p to promote chemosensitivity of RB cells via targeting SOX2.

A Novel Recombinant Virus-Like Particles Displaying B and T Cell Epitopes of Japanese Encephalitis Virus Offers Protective Immunity in Mice and Guinea Pigs.

Virus-like particles (VLPs) are non-replicative vectors for the delivery of heterologous epitopes and are considered one of the most potent inducers of cellular and humoral immune responses in mice and guinea pigs. In the present study, VLP-JEVe was constructed by the insertion of six Japanese encephalitis virus (JEV) envelope protein epitopes into different surface loop regions of PPV VP2 by the substitution of specific amino acid sequences without altering the assembly of the virus; subsequently, the protective efficacy of this VLP-JEVe was evaluated against JEV challenge in mice and guinea pigs. Mice immunized with the VLP-JEVe antigen developed high titers of neutralizing antibodies and 100% protection against lethal JEV challenge. The neutralizing and hemagglutination inhibition (HI) antibody responses were also induced in guinea pigs vaccinated with VLP-JEVe. In addition, immunization with VLP-JEVe in mice induced effective neutralizing antibodies and protective immunity against PPV (porcine parvovirus) challenge in guinea pigs. These studies suggest that VLP-JEVe produced as described here could be a potential candidate for vaccine development.

CD205+ polymorphonuclear myeloid-derived suppressor cells suppress antitumor immunity by overexpressing GLUT3.

Myeloid-derived suppressor cells (MDSCs) are responsible for antitumor immunodeficiency in tumor-bearing hosts. Primarily, MDSCs are classified into 2 groups: monocytic (M)-MDSCs and polymorphonuclear (PMN)-MDSCs. In most cancers, PMN-MDSCs (CD11b+ Ly6Clow Ly6G+ cells) represent the most abundant MDSC subpopulation. However, the functional and phenotypic heterogeneities of PMN-MDSC remain elusive, which delays clinical therapeutic targeting decisions. In the 4T1 murine tumor model, CD11b+ Ly6Glow PMN-MDSCs were sensitive to surgical and pharmacological interventions. By comprehensively analyzing 64 myeloid cell-related surface molecule expression profiles, cell density, nuclear morphology, and immunosuppressive activity, the PMN-MDSC population was further classified as CD11b+ Ly6Glow CD205+ and CD11b+ Ly6Ghigh TLR2+ subpopulations. The dichotomy of PMN-MDSCs based on CD205 and TLR2 is observed in 4T07 murine tumor models (but not in EMT6). Furthermore, CD11b+ Ly6Glow CD205+ cells massively accumulated at the spleen and liver of tumor-bearing mice, and their abundance correlated with in situ tumor burdens (with or without intervention). Moreover, we demonstrated that CD11b+ Ly6Glow CD205+ cells were sensitive to glucose deficiency and 2-deoxy-d-glucose (2DG) treatment. Glucose transporter 3 (GLUT3) knockdown by siRNA significantly triggered apoptosis and reduced glucose uptake in CD11b+ Ly6Glow CD205+ cells, demonstrating the dependence of CD205+ PMN-MDSCs survival on both glucose uptake and GLUT3 overexpression. As GLUT3 has been recognized as a target for the rescue of host antitumor immunity, our results further directed the PMN-MDSC subsets into the CD205+ GLUT3+ subpopulation as future targeting therapy.

Combined effects of mulch film-derived microplastics and atrazine on oxidative stress and gene expression in earthworm (Eisenia fetida).

With the wide use of mulch film and pesticides, mulch film-derived microplastics are very likely to produce combined effects with pesticides in agricultural soil. However, little is known about their combined toxicity on terrestrial organisms. This study aimed to investigate the combined toxicity of unused or farmland residual transparent low-density polyethylene mulch film-derived microplastics (MPs and MPs-aged, respectively) (550-1000 µm) and atrazine (ATZ; 0.02 and 2.0 mg/kg) on the earthworm (Eisenia fetida). After single and combined exposure to ATZ and microplastics for 28 d, the results showed an accumulation of reactive oxygen species, a decrease in superoxide dismutase, catalase, and glutathione-S-transferase activities, an increase in the malondialdehyde and 8-hydroxydeoxyguanosine levels, and abnormal expression of annetocin, heat shock protein 70, translationally controlled tumor protein and calreticulin genes. Integrated biological response (IBR) values calculated at the biochemical level indicated that the combined exposure to ATZ and microplastics, particularly to high concentrations of ATZ, induced greater oxidative stress in E. fetida compared with that of exposure to ATZ or microplastics alone. In addition, the IBR values calculated at the gene level did not show regular changes after combined exposure to ATZ and microplastics compared with those of a single exposure. The oxidative stress and abnormal expression of genes in E. fetida induced by MPs-aged were higher than those induced by MPs; a similar trend was observed for oxidative stress induced by MPs/MPs-aged + ATZ2.0, whereas an opposite trend was observed for the abnormal expression of genes in E. fetida induced by MPs/MPs-aged + ATZ0.02/ATZ2.0. Our results suggest that mulch film-derived microplastics have the potential to enhance the toxicity of ATZ within the soil environment.

STEAP4 Inhibits HIF-1α/PKM2 Signaling and Reduces High Glucose-Induced Apoptosis of Retinal Vascular Endothelial Cells.

BACKGROUND: Diabetic retinopathy (DR) is a vascular lesion induced by high glucose. STEAP4 is an indispensable membrane protein, which is closely related to hyperglycemic-induced cell inflammation and injury, while STEPT4 has not been studied in hyperglycemic-induced retinal vascular endothelial cell injury. METHODS: The expression of STEAP4 was detected by RT-qPCR and Western blot. CCK-8 was used to detect cell survival. STEAP4 was overexpressed by cell transfection. The expressions of cytokines TNF-α, IL-1, IL-6, ICAM-1, MDA, SOD and ROS were detected by ELISA. Cell apoptosis was detected by flow cytometry. The expressions of proteins associated with cell damage VEGF, KLF2, eNOS and apoptosis-related proteins Bax, cleaved caspase3 and Bcl2 were detected by Western blot. Finally, the expressions of HIFα and PKM2 were detected by immunofluorescence and Western blot. RESULTS: The expression of STEAP4 in hyperglycemic-induced retinal vascular endothelial cells (HRCECs) decreased gradually. Overexpression of STEAP4 reduced inflammation and apoptosis of HRCECs and improved dysfunction of them. Meanwhile, overexpression of steap4 inhibited the expression of HIF-1α/PKM2 signal. CONCLUSION: STEAP4 can be a potential therapeutic target for diabetic retinopathy by inhibiting HIF1/PKM2 signaling to reduce hyperglycemic-induced retinal cell apoptosis.

Antiproliferative effect of 2-Hydroxy-6-tridecylbenzoic acid from ginkgo biloba sarcotestas through the aryl hydrocarbon receptor pathway in triple-negative breast cancer cells.

This study aims to isolate the potential antiproliferative and cytotoxic compounds from ginkgo biloba sarcotestas (GBS) and investigates the underlying mechanism in human MDA-MB-231 and mouse 4T-1 triple-negative breast cancer cells. Our results showed that 2-Hydroxy-6-tridecylbenzoic acid was isolated by cytotoxicity-guided fractionation where different fractions were assessed using MTT assay against MDA-MB-231 and 4T-1 cells. Colony formation assay showed that 2-Hydroxy-6-tridecylbenzoic acid significantly inhibited cell proliferation. The inhibition was associated with the enhancement of cytochrome P450 (CYP) 1B1 expression in a dose- and time-dependent manner and no significant change of CYP1A1 expression by qPCR and Western blot assays in MDA-MB-231 and 4T-1 cells. The mechanism was further demonstrated by the activation of aryl hydrocarbon receptor (AhR) pathway with the upregulation of AhR, AhR nuclear translocator (ARNT) and AhR-dependent xenobiotic response elements (XRE) activity. These findings may have implications for development of anticancer agents containing 2-Hydroxy-6-tridecylbenzoic acid as functional additives.

Short-term efficacy of ticagrelor in acute ST-segment elevation myocardial infarction patients undergoing an emergency percutaneous coronary intervention.

In total, 97 acute ST-segment elevation myocardial infarction (STEMI) patients who received an emergency percutaneous coronary intervention (PCI) were enrolled and divided into a ticagrelor group and a clopidogrel group. Thrombolysis in myocardial infarction (TIMI) blood flow and the corrected TIMI frame count (CTFC) were used to assess the blood perfusion of culprit vessels. Thromboelastography (TEG) was used to evaluate the antiplatelet effect of drugs. The results showed that the incidence of TIMI grade III blood flow in the ticagrelor group was significantly higher than that in the clopidogrel group. The CTFC in the anterior descending, circumflex, and right coronary arteries was statistically significantly lower in the ticagrelor group as compared with that in the clopidogrel group. At 2 h and 7 d postdrug treatment, the adenosine diphosphate-induced platelet inhibition rate (ADP%) in the ticagrelor group increased significantly as compared with that in the clopidogrel group, and the platelet aggregation rate of the ADP pathway (MAADP) decreased significantly in the ticagrelor group versus that in the clopidogrel group. In conclusion, ticagrelor significantly improved TIMI blood flow and had a better antiplatelet effect than clopidogrel in STEMI patients undergoing an emergency PCI.

Development of a high efficient promoter finding method based on transient transfection.

In metazoan genome, the mechanism of gene expression regulation between transcriptional regulatory elements and their target gene is spatiotemporal. Active promoters possess many specific chromosomal features, such as hypersensitive to DNaseI and enrichment of specific histone modifications. In this article, we proposed a novel method which possesses a high efficiency to find promoters in vitro. A promoter-trap library was constructed with totally 706 random mouse genomic DNA fragment clones, and 260 promoter-active fragments of the library were screened by transient transfection into 4T1 cells. To demonstrate the accuracy of this promoter finding method, 13 fragments with promoter activities were randomly selected for published DNase-seq and ChIP-seq data analysis, downstream transcripts prediction and expression confirmation. qRT-PCR results showed that six predicted transcription units were successfully amplified in different mouse tissues/cells or in reconstituted mouse mammary tumors. Our results indicate that this promoter finding method can successfully detect the promoter-active fragments and their downstream transcripts.

Clinical Features of and Risk Factors for Hydrocephalus in Childhood Bacterial Meningitis.

OBJECTIVE: To explore the clinical characteristics of and analyze the risk factors for hydrocephalus in children with bacterial meningitis. METHODS: Retrospective study of a sample of children with bacterial meningitis seen on the pediatric service of Shengjing Hospital of China Medical University between January 1, 2010, and December 31, 2016. RESULTS: Overall, 9.36% (25/267) of patients presented with hydrocephalus. Among patients with hydrocephalus, the age at onset of bacterial meningitis was usually <6 months, 15 patients had confirmed bacterial etiology, and 1 patient died. The most significant results of multivariate analysis for hydrocephalus were a rural living situation, altered level of consciousness, previous treatment with antibiotics, initial cerebrospinal fluid protein >2 g/L, C-reactive protein >100 mg/L, and dexamethasone use. CONCLUSIONS: A severe clinical manifestation and significant laboratory index at admission are the most important predictors of hydrocephalus in children with bacterial meningitis.

Exploration of the immune-related signature and immune infiltration analysis for breast ductal and lobular carcinoma.

BACKGROUND: In this study, we aimed to explore the tumour associated immune signature of breast cancer (BC) and conduct integrative analyses with immune infiltrates in BC. METHODS: We downloaded the transcriptome profiling and clinical data of BC from The Cancer Genome Atlas (TCGA) database. The list of immune-related signatures was from the Innate database. The limma package was utilized to conduct the normalization, and we screened the differential immune signatures in BC. A univariate Cox regression model and the LASSO method were used to find the hub prognostic immune genes. The TAIG risk model was calculated based on the multivariate Cox regression results, and a receiver operating characteristic (ROC) curve was generated to assess the predictive power of TAIG. Moreover, we also conducted a correlation analysis between TAIG and the clinical characteristics. Additionally, we utilized the METABRIC cohort as the validation data set. The TIMER database is a comprehensive resource for performing systematic analyses of immune infiltrates across various malignancies. We evaluated the associations of immune signatures with several immune cells based on TIMER. Furthermore, we used the CIBERSORT algorithm to determine the fractions of immune cells in each sample and compared the differential distributions of immune infiltrates between two TAIG groups using the Wilcoxon rank-sum test. RESULTS: A total of 1,178 samples were obtained from the TCGA-BRCA database, but only 1,045 breast tumour samples were matched with complete transcriptome expression data. Meanwhile, we collected a total of 1,094 BC patients from the METABRIC cohort. We found a list of 1,399 differential immune signatures associated with survival, and functional analysis revealed that these genes participated in cytokine-cytokine receptor interactions, Th1 and Th2 cell differentiation and the JAK-STAT signalling pathway. The TAIG risk model was established from the multivariate Cox analysis, and we observed that high TAIG levels correlated with poor survival outcomes based on Kaplan-Meier analysis. The Kruskal-Wallis test suggested that high TAIG levels correlated with high AJCC-TNM stages and advanced pathological stages (P<0.01). We validated the well robustness of TAIG in METABRIC cohort and 5-year AUC reached up to 0.829. Moreover, we further uncovered the associations of hub immune signatures with immune cells and calculated the immune cell fractions in specific tumour samples based on gene signature expression. Last, we used the Wilcoxon rank-sum test to compare the differential immune density in the two groups and found that several immune cells had a significantly lower infiltrating density in the high TAIG groups, including CD8+ T cells (P=0.031), memory resting CD4+ T cells (P=0.026), M0 macrophages (P=0.023), and M2 macrophages (P=0.048). CONCLUSIONS: In summary, we explored the immune signature of BC and constructed a TAIG risk model to predict prognosis. Moreover, we integrated the identified immune signature with tumour-infiltrating immune cells and found adverse associations between the TAIG levels and immune cell infiltrating density.

Development of a high efficient promoter finding method based on transient transfection.

In metazoan genome, the mechanism of gene expression regulation between transcriptional regulatory elements and their target gene is spatiotemporal. Active promoters possess many specific chromosomal features, such as hypersensitive to DNaseI and enrichment of specific histone modifications. In this article, we proposed a novel method which possesses a high efficiency to find promoters in vitro. A promoter-trap library was constructed with totally 706 random mouse genomic DNA fragment clones, and 260 promoter-active fragments of the library were screened by transient transfection into 4T1 cells. To demonstrate the accuracy of this promoter finding method, 13 fragments with promoter activities were randomly selected for published DNase-seq and ChIP-seq data analysis, downstream transcripts prediction and expression confirmation. qRT-PCR results showed that six predicted transcription units were successfully amplified in different mouse tissues/cells or in reconstituted mouse mammary tumors. Our results indicate that this promoter finding method can successfully detect the promoter-active fragments and their downstream transcripts.

Construction and application of a novel genetically engineered Aspergillus oryzae for expressing proteases

Background: We aimed to test the possibility of improving polypeptide production from soybean meal fermentation by engineered Aspergillus oryzae strains. Four different protease genes were cloned and transformed into wild-type A. oryzae, and the engineered A. oryzae strains were then used for soybean meal fermentation. Results: The results showed different degrees of improvement in the protease activity of the four transformants when compared with wild-type A. oryzae. A major improvement in the polypeptide yield was achieved when these strains were used in soybean meal fermentation. The polypeptide conversion rate of one of the four transformants, A. oryzae pep, reached 35.9%, which was approximately twofold higher than that exhibited by wild-type A. oryzae. Amino acid content analysis showed that the essential amino acid content and amino acid composition of the fermentation product significantly improved when engineered A. oryzae strains were used for soybean meal fermentation. Conclusions: These findings suggest that cloning of microbial protease genes with good physicochemical properties and expressing them in an ideal host such as A. oryzae is a novel strategy to enhance the value of soybean meal.

Effect of ticagrelor on the serum level of hs-CRP, ESM-1 and short-term prognosis of patients with acute STEMI.

The aim of the present study was to observe and investigate the changes in the serum level of high-sensitivity C-reactive protein (hs-CRP), the endothelial cell-specific molecule-1 (ESM-1) and short-term prognosis of patients with ST-segment elevation myocardial infarction (STEMI) treated by ticagrelor. We enrolled 107 patients with acute STEMI who were admitted in the Department of Cardiology for the first time with occurrence of symptoms, and we successfully performed emergency operation of percutaneous coronary intervention. The patients were divided into two groups, 54 patients in the ticagrelor group (treatment group) and 53 patients in the clopidogrel group (control group), according to the administration of ticagrelor or clopidogrel in dual anti-platelet therapy. Then, we observed the changes at the time of admission, at 24 h, and 4th and 7th day after administration and investigated the correlations between them and the effect of ticagrelor on the short-term prognosis of acute STEMI patients. Significant increases of the serum levels of hs-CRP and ESM-1 were seen in patients of the two groups 24 h after administration of drugs with statistically significant differences between the groups (P<0.05), and on the 4th and 7th day we found a downward trend with statistically significant differences (P<0.05). The level of ESM-1 enhanced the increase of hs-CRP, indicating there was a positive correlation between ESM-1 and hs-CRP (r=0.535, P<0.001). A comparison of the occurrence rates of ischemic outcome event, bleeding and overall adverse events between the two groups yielded no statistically significant difference (P>0.05). In conclusion, the present study demonstrates that ticagrelor can reduce the prevalence of inflammatory reactions rapidly and stabilize the functions of vascular endothelium to improve the stability of atherosclerosis plaque and decrease the occurrence rate of thrombosis as well as ischemic outcome event without any obvious increase in the risk of bleeding. Thus, ticagrelor should be recommended in clinical practices for the treatment of patients with STEMI.