Serum cytokine biosignatures for identification of tuberculosis among HIV-positive inpatients.

BACKGROUND: Serum cytokines correlate with tuberculosis (TB) progression and are predictors of TB recurrence in people living with HIV. We investigated whether serum cytokine biosignatures could diagnose TB among HIV-positive inpatients. METHODS: We recruited HIV-positive inpatients with symptoms of TB and measured serum levels of inflammation biomarkers including IL-2, IL-4, IL-6, IL-10, tumour necrosis factor-alpha (TNF-α) and interferon-gamma (IFN-γ). We then built and tested our TB prediction model. RESULTS: 236 HIV-positive inpatients were enrolled in the first cohort and all the inflammation biomarkers were significantly higher in participants with microbiologically confirmed TB than those without TB. A binary support vector machine (SVM) model was built, incorporating the data of four biomarkers (IL-6, IL-10, TNF-α and IFN-γ). Efficacy of the SVM model was assessed in training (n=189) and validation (n=47) sets with area under the curve (AUC) of 0.92 (95% CI 0.88 to 0.96) and 0.85 (95% CI 0.72 to 0.97), respectively. In an independent test set (n=110), the SVM model yielded an AUC of 0.85 (95% CI 0.76 to 0.94) with 78% (95% CI 68% to 87%) specificity and 85% (95% CI 66% to 96%) sensitivity. Moreover, the SVM model outperformed interferon-gamma release assay (IGRA) among advanced HIV-positive inpatients irrespective of CD4+ T-cell counts, which may be an alternative approach for identifying Mycobacterium tuberculosis infection among HIV-positive inpatients with negative IGRA. CONCLUSIONS: The four-cytokine biosignature model successfully identified TB among HIV-positive inpatients. This diagnostic model may be an alternative approach to diagnose TB in advanced HIV-positive inpatients with low CD4+ T-cell counts.

An evaluation of Mp1p antigen screening for talaromycosis in HIV-infected antiretroviral therapy-naïve population in Guangdong, China.

BACKGROUND: Talaromycosis is one of the most common opportunistic infections in human immunodeficiency virus (HIV) infected patients. However, few researches have explored the prevalence in Southern China and fully assessed the value of the Mp1p antigen screening for the diagnosis of talaromycosis. METHODOLOGY/PRINCIPAL FINDINGS: We performed a cross-sectional study of HIV-infected antiretroviral therapy (ART)-naïve adult patients who were seen in 2018 at Guangzhou Eighth People's Hospital, Guangzhou Medical University. Serum samples collected from all the 784 enrolled patients were tested for Mp1p antigen using double-antibody sandwich enzyme-linked immunosorbent assay. A culture of pathogen was conducted in 350 clinically suspected patients to confirm talaromycosis. The overall prevalence of talaromycosis based on the Mp1p antigen detection was 11.4% (89/784) and peaked at 32.2% (75/233) in patients with CD4+ ≤50 Nr/µl. Logistic regression analysis found Mp1p antigen positive rate decreased with the increase in CD4+ counts (OR 0.982, 95% CI 0.977-0.987, P<0.01). The optimal cut-off point of the CD4+ count was 50 Nr/µl or less. Among the 350 patients received both fungal culture and Mp1p antigen detection, 95/350 (27.1%) patients were culture-positive for a Talaromyces marneffei, 75/350 (21.4%) patients were Mp1p antigen positive. The Mp1p antigen assay showed a good agreement to the culture of pathogen, and the sensitivity, specificity, positive predictive value, negative predictive value and kappa value was 71.6% (68/95), 97.3% (248/255), 90.7% (68/75), 90.2% (248/275), and 0.737, respectively. The screening accuracy of the Mp1p antigen assay in patients with CD4+ counts of ≤50 Nr/µl was superior to that in those with higher CD4+ counts. CONCLUSIONS/SIGNIFICANCE: Mp1p antigen screening can be an effective tool for more efficient diagnosis of Talaromycosis, especially in HIV/AIDS patients with low CD4+ counts. Future validation studies are needed.

Immunosensor with Enhanced Electrochemiluminescence Signal Using Platinum Nanoparticles Confined within Nanochannels for Highly Sensitive Detection of Carcinoembryonic Antigen.

Rapid, highly sensitive, and accurate detection of tumor biomarkers in serum is of great significance in cancer screening, early diagnosis, and postoperative monitoring. In this study, an electrochemiluminescence (ECL) immunosensing platform was constructed by enhancing the ECL signal through in situ growth of platinum nanoparticles (PtNPs) in a nanochannel array, which can achieve highly sensitive detection of the tumor marker carcinoembryonic antigen (CEA). An inexpensive and readily available indium tin oxide (ITO) glass electrode was used as the supporting electrode, and a layer of amino-functionalized vertically ordered mesoporous silica film (NH2-VMSF) was grown on its surface using an electrochemically assisted self-assembly method (EASA). The amino groups within the nanochannels served as anchoring sites for the one-step electrodeposition of PtNPs, taking advantage of the confinement effect of the ultrasmall nanochannels. After the amino groups on the outer surface of NH2-VMSF were derivatized with aldehyde groups, specific recognition antibodies were covalently immobilized followed by blocking nonspecific binding sites to create an immunorecognition interface. The PtNPs, acting as nanocatalysts, catalyzed the generation of reactive oxygen species (ROS) with hydrogen peroxide (H2O2), significantly enhancing the ECL signal of the luminol. The ECL signal exhibited high stability during continuous electrochemical scanning. When the CEA specifically bound to the immunorecognition interface, the resulting immune complexes restricted the diffusion of the ECL emitters and co-reactants towards the electrode, leading to a reduction in the ECL signal. Based on this immune recognition-induced signal-gating effect, the immunosensor enabled ECL detection of CEA with a linear range of 0.1 pg mL-1 to 1000 ng mL-1 with a low limit of detection (LOD, 0.03 pg mL-1). The constructed immunosensor demonstrated excellent selectivity and can achieve CEA detection in serum.

Research Note: Dietary resveratrol supplementation improves the hepatic antioxidant capacity and attenuates lipopolysaccharide-induced inflammation in yellow-feathered broilers.

This experiment investigated the protective effect of resveratrol (RES) on the hepatic antioxidant status and systemic inflammation in yellow-feathered broilers challenged with lipopolysaccharide (LPS). A total of 240 healthy 1-day-old yellow-feathered broilers were randomly divided into 4 groups (control, LPS, RES, and RES+LPS), with 5 replicates of 12 chickens per replicate. The experiment lasted 21 d. The broilers were fed with either the basal diet or the basal diet supplemented with 400 mg/kg RES followed by intraperitoneal challenge with LPS (1 mg/kg body weight) or the same amount of saline at d 16, 18, and 20. The results showed that dietary RES supplementation could improve the activities of total antioxidant capacity (T-AOC) and superoxide dismutase (SOD) in the liver of yellow-feathered broilers challenged with LPS (P < 0.05). Furthermore, LPS challenge increased the plasma interleukin-17 (IL-17) concentration, the hepatic interleukin-6 (IL-6) and interleukin-1ß (IL-1ß) concentrations, as well as the concentrations of tumor necrosis factor (TNF-α), IL-6, and IL-1ß in the spleen (P < 0.05), and decreased the transforming growth factor-ß (TGF-ß) concentrations in the plasma, liver, and spleen (P < 0.05). However, dietary RES supplementation could reduce the increased TNF-α levels in the plasma, liver, and spleen induced by LPS, and increased TGF-ß level in the liver and spleen (P < 0.05). Collectively, these results suggest that dietary RES supplementation could effectively improve the hepatic antioxidant capacity and attenuate LPS-induced inflammation in yellow-feathered broilers during the starter stage.

A simple electrochemical immunosensor based on a chitosan/reduced graphene oxide nanocomposite for sensitive detection of biomarkers of malignant melanoma.

The sensitive and specific detection of tumor biomarkers is crucial for early diagnosis and treatment of malignant melanoma. Immunoassay with a simple sensing interface and high sensitivity is highly desirable. In this work, a simple electrochemical immunosensor based on a chitosan/reduced graphene oxide (CS-rGO) nanocomposite was developed for sensitive determination of an S-100B protein, a tumor marker of malignant melanoma. CS-rGO nanocomposite were prepared by chemical reduction of graphene oxide in the presence of chitosan and modified on glassy carbon electrode (GCE) to provide a biofriendly, conductive, and easily chemically modified matrix for further immobilization of antibodies. Anti-S-100B antibodies were grafted onto the chitosan molecules to fabricate the immunorecognition interface by a simple glutaraldehyde cross-linking method. Electrochemical determination of S-100B was achieved by measuring the decreased current signal of solution phase electrochemical probes, which originated from the increased steric hindrance and insulation caused by the formation of antigen-antibody complexes at the electrode interface. Due to the good conductivity, high surface area, excellent biocompatibility, and good film-forming ability of CS-rGO, the constructed immunosensor exhibited good stability, high selectivity and sensitivity, a wide dynamic range from 10 fg mL-1 to 1 ng mL-1 and a low limit of detection of 1.9 pg mL-1 (S/N = 3). Moreover, the sensor was also applicable for the sensitive detection of S-100B protein in real human serum samples.

Core immune cell infiltration signatures identify molecular subtypes and promote precise checkpoint immunotherapy in cutaneous melanoma.

Yutao Wang, China Medical University, ChinaThe tumor microenvironment (TME) has been shown to impact the prognosis of tumors in patients including cutaneous melanoma (CM); however, not all components of TME are important. Given the aforementioned situation, the functional immune cell contents correlated with CM patient prognosis are needed to optimize present predictive models and reflect the overall situation of TME. We developed a novel risk score named core tumor-infiltrating immune cell score (cTICscore), which showed certain advantages over existing biomarkers or TME-related signatures in predicting the prognosis of CM patients. Furthermore, we explored a new gene signature named cTILscore-related module gene score (cTMGs), based on four identified TME-associated genes (GCH1, GZMA, PSMB8, and PLAAT4) showing a close correlation with the cTICscore, which was generated by weighted gene co-expression network analysis and least absolute shrinkage and selection operator analysis to facilitate clinical application. Patients with low cTMGs had significantly better overall survival (OS, P = 0.002,< 0.001, = 0.002, and = 0.03, respectively) in the training and validating CM datasets. In addition, the area under the curve values used to predict the immune response in four CM cohorts were 0.723, 0.723, 0.754, and 0.792, respectively, and that in one gastric cohort was 0.764. Therefore, the four-gene signature, based on cTICscore, might improve prognostic information, serving as a predictive tool for CM patients receiving immunotherapy.cutaneous melanoma, tumor microenvironment, prognosis, immunotherapy, cTICscore.

Effect of dietary resveratrol supplementation on growth performance, antioxidant capacity, intestinal immunity and gut microbiota in yellow-feathered broilers challenged with lipopolysaccharide.

Resveratrol (RES) displays strong antioxidant and anti-inflammatory properties in protecting the animals from various stressors and inflammatory injuries, but its interrelationship with the gut microbiota remained largely unclear. This study was carried out to investigate the effects of dietary RES supplementation on growth performance, antioxidant capacity, intestinal immunity and gut microbiota in yellow-feathered broilers challenged by lipopolysaccharide (LPS). A total of 240 yellow-feathered broilers were randomly assigned to four treatment groups in a 2 × 2 factorial design. The broilers were fed with the control diet or control diet supplemented with 400 mg/kg RES, followed by challenge with LPS or the same amount of saline. Dietary RES supplementation significantly alleviated the decreases in the final body weight (BW), average daily gain (ADG), and ADFI induced by LPS (P < 0.05). LPS challenge significantly increased plasma concentrations of triglyceride, high-density lipoprotein cholesterol (HDL-C), aspartate aminotransferase (AST), and cortisol levels, but decreased triiodothyronine (T3) and insulin levels (P < 0.05). Dietary supplementation with RES significantly reversed the elevated creatinine concentrations and the decreased concentrations of T3 and insulin caused by LPS (P < 0.05). Moreover, dietary RES supplementation significantly increased plasma total antioxidant capacity (T-AOC) and catalase (CAT) activities and superoxide dismutase (SOD) and T-AOC activities in jejunal mucosa and reduced malondialdehyde (MDA) concentration in the plasma (P < 0.05). The reduction in the villus height to crypt depth ratio in duodenum, jejunum and ileum and the shortening of villus height in jejunum and ileum caused by LPS were also alleviated by RES treatment (P < 0.05). Furthermore, the increased concentrations of intestinal tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and IL-1ß caused by LPS were significantly decreased by RES treatment (P < 0.05). Dietary RES treatment increased the mRNA expression of claudin-1, claudin-5, occludin, and zonula occludens-1 (ZO-1), and decreased mRNA expression of IL-1ß, IL-8, IL-17, and TNF-α after LPS challenge (P < 0.05). Dietary RES treatments significantly decreased the dominance of cecal microbiota, and increased the Pieiou-e and Simpson index. Moreover, dietary RES supplementation increased relative abundance of UCG_ 009, Erysipelotrichaceae, Christensenellaceae_R-7_group, Anaerotruncus, RF39, and Ruminococcus while decreasing the abundance of Alistipes at genus level. Spearman correlation analysis revealed that the microbes at the order and genus levels significantly correlated with indicators of growth performance, antioxidant capacity, and intestinal health. Collectively, dietary supplementation with 400 mg/kg RES could improve growth performance and antioxidant capacity, and modulate intestinal immunity in yellow-feathered broilers challenged by LPS at early stage, which might be closely associated with the regulation of gut microbiota community composition.

Effects of Dietary L-Theanine on Growth Performance, Antioxidation, Meat Quality, and Intestinal Microflora in White Feather Broilers With Acute Oxidative Stress.

In order to reduce the negative effects caused by oxidative stress on broilers, it is particularly important to find ways to alleviate oxidative stress. As a natural plant extract, L-theanine has a variety of biological effects, such as improving antioxidant capacity, promoting growth, and enhancing immunity and antitumor. This trial evaluated the effects of dietary supplementation of L-theanine on growth performance, antioxidation, meat quality, and intestinal microflora in 817 White Feather Broilers. A total of 108 21-day-old 817 broilers with similar body weight (BW) were randomly divided into three groups with six replicates per group and six chickens within each replicate. The three groups were corn-soybean-based diet (NC group); basal diet plus drinking water with 30 mg hydrocortisone/kg (PC group); and basal diet supplemented with 400 mg L-theanine/kg plus drinking water with 30 mg hydrocortisone/kg (LT group). Compared with the NC group, from 21 to 24 days of age, the PC and LT groups had decreased BW, average daily gain (ADG), and average daily feed intake (ADFI), and increased feed to gain ratio (F/G; p < 0.05). At 24 days of age, the LT group had improved superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activities in serum as compared to the NC group (p < 0.05). The LT group broilers also had significantly higher concentrations of malondialdehyde (MDA) in serum and liver (p < 0.05). On the 42nd days, the PC group had lower PH45min (p < 0.05) than the NC and LT groups and higher cooking loss and shear force (p < 0.05). Moreover, the villi height of the PC group was significantly lower in jejunum than the NC group (p < 0.05). The LT group had a higher ZO-1 content in duodenum than the NC and PC groups (p < 0.05). The activity of GSH-Px in the liver of the LT group was increased than in the PC group (p < 0.05). The relative abundance of Firmicutes in the LT group was significantly higher than in the NC and PC groups (p < 0.05). These results suggested that the effects of acute oxidative stress on growth performance and meat quality of broilers are continuous, and dietary supplementation of L-theanine could improve the growth performance and meat quality, enhance the intestinal mucosal barrier and antioxidant capacity, and improve the composition of the intestinal flora of broilers caused by acute oxidative stress.

[Clinical features of children with colorectal polyps and the efficacy of endoscopic treatment: an analysis of 1 351 cases]. / å„¿ç«¥ç»“ç›´è‚ æ¯è‚‰1 351ä¾‹çš„ä¸´åºŠç‰¹å¾åŠå† é•œä¸‹æ²»ç–—æ•ˆæžœåˆ†æž.

OBJECTIVES: To study the clinical features of children with colorectal polyps and the efficacy of endoscopic treatment. METHODS: A retrospective analysis was performed on the medical data of 1 351 children with colorectal polyps who were admitted and received colonoscopy and treatment in the past 8 years, including clinical features and the pattern and outcomes of endoscopic treatment. RESULTS: Among the 1 351 children, 893 (66.10%) were boys and 981 (72.61%) had an age of 2-<7 years, and hematochezia (1 307, 96.74%) was the most common clinical manifestation. Of all the children, 89.27% (1 206/1 351) had solitary polyps, and 95.77% (1 290/1 347) had juvenile polyps. The polyps were removed by electric cauterization with hot biopsy forceps (6 cases) or high-frequency electrotomy and electrocoagulation after snare ligation (1 345 cases). A total of 1 758 polyps were resected, among which 1 593 (90.61%) were pedunculated and 1 349 (76.73%) had a diameter of <2 cm. Postoperative complications included bleeding in 51 children (3.77%), vomiting in 87 children (6.44%), abdominal pain in 14 children (1.04%), and fever in 39 children (2.89%), while no perforation was observed. The children aged <3 years had the highest incidence rates of postoperative bleeding and fever (P<0.0125), and the children with a polyp diameter of ≥2 cm had significantly higher incidence rates of postoperative bleeding, vomiting, and fever (P<0.05). CONCLUSIONS: Solitary polyps, pedunculated polyps, and juvenile polyps are common types of pediatric colorectal polyps. Electric cauterization with hot biopsy forceps or high-frequency electrotomy and electrocoagulation after snare ligation can effectively remove colorectal polyps in children, with good efficacy and few complications. Younger age and larger polyp diameter are associated with a higher risk of postoperative bleeding.

Protective Effects of Lactobacillus plantarum Lac16 on Clostridium perfringens Infection-Associated Injury in IPEC-J2 Cells.

Clostridium perfringens (C. perfringens) causes intestinal injury through overgrowth and the secretion of multiple toxins, leading to diarrhea and necrotic enteritis in animals, including pigs, chickens, and sheep. This study aimed to investigate the protective effects of Lactobacillus plantarum (L. plantarum) Lac16 on C. perfringens infection-associated injury in intestinal porcine epithelial cell line (IPEC-J2). The results showed that L. plantarum Lac16 significantly inhibited the growth of C. perfringens, which was accompanied by a decrease in pH levels. In addition, L. plantarum Lac16 significantly elevated the mRNA expression levels of host defense peptides (HDPs) in IPEC-J2 cells, decreased the adhesion of C. perfringens to IPEC-J2 cells, and attenuated C. perfringens-induced cellular cytotoxicity and intestinal barrier damage. Furthermore, L. plantarum Lac16 significantly suppressed C. perfringens-induced gene expressions of proinflammatory cytokines and pattern recognition receptors (PRRs) in IPEC-J2 cells. Moreover, L. plantarum Lac16 preincubation effectively inhibited the phosphorylation of p65 caused by C. perfringens infection. Collectively, probiotic L. plantarum Lac16 exerts protective effects against C. perfringens infection-associated injury in IPEC-J2 cells.

Differential DNA Methylation and Gene Expression Between ALV-J-Positive and ALV-J-Negative Chickens.

Background: Avian leukosis virus subgroup J (ALV-J) is an oncogenic virus that causes serious economic losses in the poultry industry; unfortunately, there is no effective vaccine against ALV-J. DNA methylation plays a crucial role in several biological processes, and an increasing number of diseases have been proven to be related to alterations in DNA methylation. In this study, we screened ALV-J-positive and -negative chickens. Subsequently, we generated and provided the genome-wide gene expression and DNA methylation profiles by MeDIP-seq and RNA-seq of ALV-J-positive and -negative chicken samples; 8,304 differentially methylated regions (DMRs) were identified by MeDIP-seq analysis (p ≤ 0.005) and 515 differentially expressed genes were identified by RNA-seq analysis (p ≤ 0.05). As a result of an integration analysis, we screened six candidate genes to identify ALV-J-negative chickens that possessed differential methylation in the promoter region. Furthermore, TGFB2 played an important role in tumorigenesis and cancer progression, which suggested TGFB2 may be an indicator for identifying ALV-J infections.

Co-Cropping Indian Mustard and Silage Maize for Phytoremediation of a Cadmium-Contaminated Acid Paddy Soil Amended with Peat.

Co-cropping is an eco-friendly strategy to improve the phytoremediation capacity of plants growing in soils contaminated with heavy metals such as cadmium (Cd). This study was conducted to investigate the effects of co-cropping Indian mustard (Brassicajuncea) and silage maize (Zeamays) and applying peat on the phytoremediation of a Cd-contaminated acid paddy soil via characterizing plant growth and Cd uptake in pot experiments. There were six planting patterns (Control: no plants; MI-2 and MI-4: mono-cropping of Indian mustard at low and high densities, respectively; MS: mono-cropping of silage maize; CIS-2 and CIS-4: co-cropping of Indian mustard at low and high densities with silage maize, respectively) and two application rates of peat (NP: 0; WP: 30 g kg-1). When Indian mustard and silage maize were co-cropped, the shoot biomass of Indian mustard plants per pot was significantly (p < 0.05) lower than that obtained in the mono-cropping systems, with a substantial reduction (55-72%) in the same plant density group. The shoot biomass of silage maize plants in the mono-cropping systems did not differ significantly from that in the co-cropping systems regardless of the density of Indian mustard. The growth-promoting effect of the peat application was more pronounced in Indian mustard than silage maize. Under the low density of Indian mustard, the co-cropping systems significantly (p < 0.05) decreased Cd uptake by silage maize. Additionally, soil amendment with peat significantly (p < 0.05) increased shoot Cd removal rate and Cd translocation factor value in the co-cropping systems. Taken together, the results demonstrated that silage maize should be co-cropped with Indian mustard at an appropriate density in Cd-polluted soils to achieve simultaneous remediation of Cd-contaminated soils (via Indian mustard) and production of crops (here, silage maize). Peat application was shown to promote the removal of Cd from soil and translocation of Cd into shoots and could contribute to enhanced phytoremediation of Cd-contaminated acid paddy soil.

Hypoxanthine Induces Muscular ATP Depletion and Fatigue via UCP2.

Hypoxanthine (Hx), an intermediate metabolite of the purine metabolism pathway which is dramatically increased in blood and skeletal muscle during muscle contraction and metabolism, is characterized as a marker of exercise exhaustion. However, the physiological effects of Hx on skeletal muscle remain unknown. Herein, we demonstrate that chronic treatment with Hx through dietary supplementation resulted in skeletal muscle fatigue and impaired the exercise performance of mice without affecting their growth and skeletal muscle development. Hx increased the uncoupling protein 2 (UCP2) expression in the skeletal muscle, which led to decreased energy substrate storage and enhanced glycolysis. These effects could also be verified in acute treatment with Hx through intraperitoneal injection. In addition, muscular specifically knockout of UCP2 through intra-muscle tissue injection of adenovirus-associated virus reversed the effects of Hx. In conclusion, we identified a novel role of Hx in the skeletal muscular fatigue mediated by UCP2-dependent mitochondrial uncoupling. This finding may shed light on the pathological mechanism of clinical muscle dysfunctions due to abnormal metabolism, such as muscle fatigue and weakness.

Identification and analysis of immune-related subtypes of hepatocellular carcinoma.

Hepatocellular carcinoma is a malignance that remains difficult to cure. Immunotherapy has shown its potential application in a variety of refractory malignancies. Due to the complexity of immune microenvironment of hepatocellular carcinoma, the efficacy of immunotherapy for hepatocellular carcinoma is not as effective as expected. Expression data of hepatocellular carcinoma from the TCGA and ICGC databases were used for classification and verification of hepatocellular carcinoma subtypes. The immune-related functions and pathways were identified via gene set enrichment analysis, while the sections denoting the subsets of the immune cells were estimated using the CIBERSORT algorithm. Immunity low (Immunity_L), immunity medium (Immunity_M), and immunity high (Immunity_H) were specified as the three immune-related subtypes of hepatocellular carcinoma. The quantity of stromal and immune cells was the most substantial in Immunity_H, compared to the other subtypes. Interestingly, the proportion of M0 macrophages decreased from Immunity_L to Immunity_H, while the proportion of CD8 T cells increased. Furthermore, the HLA genes expression levels, as well as those of six immune checkpoint genes were substantially lower in Immunity_L than in Immunity_H. Functional analysis was performed for 1512 differentially expressed genes between Immunity_L and Immunity_H. Finally, the PPI network was constructed with 118 nodes. The highest connectivity degree nodes were B2M, HLA-DRA, and HLA-DRB1. The above results were verified in ICGC-JP and ICGC-FR databases with a consistent trend. In this study, we divided hepatocellular carcinoma into three subtypes and explored the immune-related characteristics of these subtypes. These results may provide new insights for immunotherapy of hepatocellular carcinoma.

Genes regulating membrane-associated E-cadherin and proliferation in adenomatous polyposis coli mutant colon cancer cells: High content siRNA screen.

Truncating mutations in the tumour suppressor gene APC occur frequently in colorectal cancers and result in the deregulation of Wnt signalling as well as changes in cell-cell adhesion. Using quantitative imaging based on the detection of membrane-associated E-cadherin, we undertook a protein coding genome-wide siRNA screen to identify genes that regulate cell surface E-cadherin in the APC-defective colorectal cancer cell line SW480. We identified a diverse set of regulators of E-cadherin that offer new insights into the regulation of cell-cell adhesion, junction formation and genes that regulate proliferation or survival of SW480 cells. Among the genes whose depletion promotes membrane-associated E-cadherin, we identified ZEB1, the microRNA200 family, and proteins such as a ubiquitin ligase UBE2E3, CDK8, sorting nexin 27 (SNX27) and the matrix metalloproteinases, MMP14 and MMP19. The screen also identified 167 proteins required for maintaining E-cadherin at cell-cell adherens junctions, including known junctional proteins, CTNND1 and CTNNA1, as well as signalling enzymes, DUSP4 and MARK2, and transcription factors, TEAD3, RUNX2 and TRAM2. A better understanding of the post-translational regulation of E-cadherin provides new opportunities for restoring cell-cell adhesion in APC-defective cells.

Identification of eight-protein biosignature for diagnosis of tuberculosis.

BACKGROUND: Biomarker-based tests for diagnosing TB currently rely on detecting Mycobacterium tuberculosis (Mtb) antigen-specific cellular responses. While this approach can detect Mtb infection, it is not efficient in diagnosing TB, especially for patients who lack aetiological evidence of the disease. METHODS: We prospectively enrolled three cohorts for our study for a total of 630 subjects, including 160 individuals to screen protein biomarkers of TB, 368 individuals to establish and test the predictive model and 102 individuals for biomarker validation. Whole blood cultures were stimulated with pooled Mtb-peptides or mitogen, and 640 proteins within the culture supernatant were analysed simultaneously using an antibody-based array. Sixteen candidate biomarkers of TB identified during screening were then developed into a custom multiplexed antibody array for biomarker validation. RESULTS: A two-round screening strategy identified eight-protein biomarkers of TB: I-TAC, I-309, MIG, Granulysin, FAP, MEP1B, Furin and LYVE-1. The sensitivity and specificity of the eight-protein biosignature in diagnosing TB were determined for the training (n=276), test (n=92) and prediction (n=102) cohorts. The training cohort had a 100% specificity (95% CI 98% to 100%) and 100% sensitivity (95% CI 96% to 100%) using a random forest algorithm approach by cross-validation. In the test cohort, the specificity and sensitivity were 83% (95% CI 71% to 91%) and 76% (95% CI 56% to 90%), respectively. In the prediction cohort, the specificity was 84% (95% CI 74% to 92%) and the sensitivity was 75% (95% CI 57% to 89%). CONCLUSIONS: An eight-protein biosignature to diagnose TB in a high-burden TB clinical setting was identified.

Detect molecular spectral features of newly developed Vicia faba varieties and protein metabolic characteristics in ruminant system using advanced synchrotron radiation based infrared microspectroscopy: A preliminary study.

Recently, advanced synchrotron radiation-based Fourier transform infrared microspectroscopy (SR-IMS) has been developed as a rapid, direct, non-destructive and bioanalytical technique. To date, there has been very little application of this technique to study the molecular structure make-up in pulse seeds. Thus, the objectives of this study were to detect the interactive association between protein molecular structure and nutrient availability of newly developed Vicia faba varieties. Two different varieties of faba beans (CDC Snowdrop = low-tannin variety; vs. FB9-4 = high-tannin variety) were selected for this study. The molecular spectra data were collected by using SR-IMS. The ratio of both amide I to II area and height were higher (P < 0.01), while the ratio of α-helix to ß-sheet was lower (P < 0.05) in CDC Snowdrop compared to FB9-4. The crude protein (CP) content and the predicted truly digestible nutrients as well as the bioenergy values did not vary between two varieties. The CDC Snowdrop had exhibited a higher (P < 0.01) rapidly degradable CP fraction (75.99 vs. 71.45% on CP) and a lower (P < 0.01) moderately degradable CP fraction (19.43 vs. 22.85% on CP), resulting in a higher (P < 0.01) rumen degradable protein and a lower (P < 0.01) rumen undegradable protein content than that of FB9-4 variety. However, the total supply of digestible rumen undegraded feed protein was higher (P < 0.05) in FB9-4 than CDC Snowdrop. Strong positive correlations were found between the ratio of α-helix to ß-sheet and CP contents (R = 0.86, P < 0.01) as well as the truly digestible CP contents (R = 0.83, P < 0.01); respectively. In conclusion, the results of this study reveal that the protein are metabolized differently between different type of faba bean varieties and the advanced SR-IMS molecular spectroscopy can be used to rapidly delineate protein molecular structure motifs along with their nutritive value in ruminant livestock system.

The correlation and prognostic value of serum levels of soluble programmed death protein 1 (sPD-1) and soluble programmed death-ligand 1 (sPD-L1) in patients with hepatocellular carcinoma.

BACKGROUND: Blocking the programmed death protein 1 (PD-1)/programmed death-ligand 1 (PD-L1) pathway in hepatocellular carcinoma (HCC) is a very promising approach in immunotherapy. However, the correlation and prognostic values of serum soluble PD-1 and PD-L1 (sPD-1/sPD-L1) have not been explored conjointly in HCC patients. METHODS: This study retrospectively included 120 HCC patients receiving radical resection. The serum levels of sPD-1/sPD-L1 and inflammatory cytokines were measured by antibody array assay. Immunohistochemistry was applied to assess both the expression of membrane-bound PD-L1, and the number of CD4+ tumor-infiltrating lymphocytes (TILs) and CD8+ TILs. RESULTS: The best cut-off values of sPD-1 and sPD-L1 for predicting disease-free survival (DFS) were 33.0 µg/ml and 11.2 µg/ml, respectively. Multivariable analysis showed that sPD-L1 was a negative independent prognostic factor [DFS, Hazard Ratio (HR) 2.58, 95% CI 1.14-5.84, P = 0.023; overall survival (OS), HR 1.77, 95% CI 1.01-3.12, P = 0.048], while sPD-1 was a favorable independent prognostic factor (DFS, HR 0.32, 95% CI 0.14-0.74, P = 0.007; OS, HR 0.54, 95% CI 0.30-0.98, P = 0.044) in HCC patients. We also observed some similar associations between inflammatory cytokines (IL-10, IL-17, TNF-α) and sPD-1 or sPD-L1, as well as a close positive association between sPD-1 and sPD-L1. No significant associations of sPD-1/sPD-L1 with either intra-tumoral PD-L1 expression, or the numbers of CD4+ TILs and CD8+ TILs were determined. CONCLUSIONS: Our findings indicate that sPD-1 and sPD-L1 are independent prognostic factors with opposite prognostic roles in predicting both DFS and OS in HCC patients.

Urokinase plasminogen activator secreted by cancer-associated fibroblasts induces tumor progression via PI3K/AKT and ERK signaling in esophageal squamous cell carcinoma.

Cancer-associated fibroblasts (CAFs) are believed to influence tumor behavior and clinical outcomes. We previously showed that conditioned medium (CM) from CAFs induces proliferation and motility of esophageal squamous cell carcinoma (ESCC) cells. Here, we investigated the molecular mechanisms by which the CAF-secreted proteins induce ESCC development and progression. Using antibody arrays, we identified urokinase plasminogen activator (uPA) as one of the main proteins whose release was increased in CAFs compared to normal fibroblasts (NFs). Immunohistochemical analysis of pathological sections showed that uPA-positive cells were localized at the boundaries of tumor and stroma tissues, in stroma between tumor nests, and within the tumors. Increased stromal uPA levels (132/146 cases) correlated with tumor invasion (p < 0.05) and overall survival of ESCC patients (p < 0.05). In vitro assays showed that uPA promotes ESCC cell proliferation, migration, and invasion via PI3K/AKT and ERK signaling pathways. In vivo, anti-uPA antibody suppressed tumor growth in ESCC xenografts. These results suggest that uPA released from stroma, and especially from CAFs, might be a predictive marker for ESCC diagnosis and prognosis, as well as an effective therapeutic target.

A retrospective analysis of the clinicopathological characteristics of large cell carcinoma of the lung.

The aim of the present study was to analyze and summarize the clinicopathological characteristics of large-cell lung carcinoma (LCLC) of the lung, in order to improve the definite diagnosis rate of LCLC. Clinicopathological data of 174 patients with LCLC, confirmed pathologically, were retrospectively reviewed. The 174 cases of LCLC accounted for 5.7% of the total lung cancer cases during the corresponding time period at the Affiliated Cancer Hospital of Tianjin Medical University (Tianjin, China), among which there were 131 males and 43 females with an average age of 61.4 years. The postoperative pathological diagnosis of the 174 cases showed 80 cases of classic LCLC, 64 cases of large cell neuroendocrine carcinoma (LCNEC), six cases of combined LCNEC, 19 cases of basaloid carcinoma, three cases of clear cell carcinoma and two cases of lymphoepithelioma-like carcinoma. Of the total 174 LCLC cases, 96 patients exhibited lymph node metastasis. LCLC is a highly aggressive malignancy with a high tendency of invasion and metastasis, although the incidence rate is low. A definite diagnosis of LCLC primarily relies on the pathological diagnosis. Each subtype of LCLC has its own pathomorphological and immunohistochemical characteristics.