Multiomics profiling reveals the benefits of gamma-delta (γδ) T lymphocytes for improving the tumor microenvironment, immunotherapy efficacy and prognosis in cervical cancer.

BACKGROUND: As an unconventional subpopulation of T lymphocytes, γδ T cells can recognize antigens independently of major histocompatibility complex restrictions. Recent studies have indicated that γδ T cells play contrasting roles in tumor microenvironments-promoting tumor progression in some cancers (eg, gallbladder and leukemia) while suppressing it in others (eg, lung and gastric). γδ T cells are mainly enriched in peripheral mucosal tissues. As the cervix is a mucosa-rich tissue, the role of γδ T cells in cervical cancer warrants further investigation. METHODS: We employed a multiomics strategy that integrated abundant data from single-cell and bulk transcriptome sequencing, whole exome sequencing, genotyping array, immunohistochemistry, and MRI. RESULTS: Heterogeneity was observed in the level of γδ T-cell infiltration in cervical cancer tissues, mainly associated with the tumor somatic mutational landscape. Definitely, γδ T cells play a beneficial role in the prognosis of patients with cervical cancer. First, γδ T cells exert direct cytotoxic effects in the tumor microenvironment of cervical cancer through the dynamic evolution of cellular states at both poles. Second, higher levels of γδ T-cell infiltration also shape the microenvironment of immune activation with cancer-suppressive properties. We found that these intricate features can be observed by MRI-based radiomics models to non-invasively assess γδ T-cell proportions in tumor tissues in patients. Importantly, patients with high infiltration levels of γδ T cells may be more amenable to immunotherapies including immune checkpoint inhibitors and autologous tumor-infiltrating lymphocyte therapies, than to chemoradiotherapy. CONCLUSIONS: γδ T cells play a beneficial role in antitumor immunity in cervical cancer. The abundance of γδ T cells in cervical cancerous tissue is associated with higher response rates to immunotherapy.

Dosimetric analysis of brachial plexopathy after stereotactic body radiotherapy: Significance of organ delineation.

OBJECTIVES: Examine the significance of contouring the brachial plexus (BP) for toxicity estimation and select metrics for predicting radiation-induced brachial plexopathy (RIBP) after stereotactic body radiotherapy. MATERIALS AND METHODS: Patients with planning target volume (PTV) ≤ 2 cm from the BP were eligible. The BP was contoured primarily according to the RTOG 1106 atlas, while subclavian-axillary veins (SAV) were contoured according to RTOG 0236. Apical PTVs were classified as anterior (PTV-A) or posterior (PTV-B) PTVs. Variables predicting grade 2 or higher RIBP (RIBP2) were selected through least absolute shrinkage and selection operator regression and logistic regression. RESULTS: Among 137 patients with 140 BPs (median follow-up, 32.1 months), 11 experienced RIBP2. For patients with RIBP2, the maximum physical dose to the BP (BP-Dmax) was 46.5 Gy (median; range, 35.7 to 60.7 Gy). Of these patients, 54.5 % (6/11) satisfied the RTOG limits when using SAV delineation; among them, 83.3 % (5/6) had PTV-B. For patients with PTV-B, the maximum physical dose to SAV (SAV-Dmax) was 11.2 Gy (median) lower than BP-Dmax. Maximum and 0.3 cc biologically effective doses to the BP based on the linear-quadratic-linear model (BP-BEDmax LQL and BP-BED0.3cc LQL, α/ß = 3) were selected as predictive variables with thresholds of 118 and 73 Gy, respectively. CONCLUSION: Contouring SAV may significantly underestimate the RIBP2 risk in dosimetry, especially for patients with PTV-B. BP contouring indicated BP-BED0.3cc LQL and BP-BEDmax LQL as potential predictors of RIBP2.

Can low-dose intravenous bevacizumab be as effective as high-dose bevacizumab for cerebral radiation necrosis?

Although intravenous bevacizumab (IVBEV) is the most promising treatment for cerebral radiation necrosis (CRN), there is no conclusion on the optimal dosage. Our retrospective study aimed to compare the efficacy and safety of high-dose with low-dose IVBEV in treating CRN associated with radiotherapy for brain metastases (BMs). This paper describes 75 patients who were diagnosed with CRN secondary to radiotherapy for BMs, treated with low-dose or high-dose IVBEV and followed up for a minimum of 6 months. The clinical data collected for this study include changes in brain MRI, clinical symptoms, and corticosteroid usage before, during, and after IVBEV treatment. At the 3-month mark following administration of IVBEV, a comparison of two groups revealed that the median percentage decreases in CRN volume on T2-weighted fluid-attenuated inversion recovery and T1-weighted gadolinium contrast-enhanced image (T1CE), as well as the signal ratio reduction on T1CE, were 65.8% versus 64.8% (p = 0.860), 41.2% versus 51.9% (p = 0.396), and 37.4% versus 35.1% (p = 0.271), respectively. Similarly, at 6 months post-IVBEV, the median percentage reductions of the aforementioned parameters were 59.5% versus 62.0% (p = 0.757), 39.1% versus 31.3% (p = 0.851), and 35.4% versus 28.2% (p = 0.083), respectively. Notably, the incidence of grade ≥3 adverse events was higher in the high-dose group (n = 4, 9.8%) than in the low-dose group (n = 0). Among patients with CRN secondary to radiotherapy for BMs, the administration of high-dose IVBEV did not demonstrate superiority over low-dose IVBEV. Moreover, the use of high-dose IVBEV was associated with a higher incidence of grade ≥3 adverse events compared with low-dose IVBEV.

Impact of radiation dose to the immune system on disease progression and survival for early-stage non-small cell lung cancer treated with stereotactic body radiation therapy.

OBJECTIVES: Although the effects of estimated dose of radiation to immune cells (EDRIC) in stage III NSCLC, LA-NSCLC, LS-SCLC and esophageal cancer on clinical outcomes have been studied, its impact in early-stage non-small cell lung cancer (ES-NSCLC) is unknown. In this study, we evaluated the role of EDRIC and identified the factors influencing EDRIC in this population. METHODS AND MATERIALS: We retrospectively analyzed 211 pathologically confirmed ES-NSCLC patients who were treated with SBRT between 2007 and 2020. EDRIC was calculated based on the model developed by Jin et al. and improved by Ladbury et al. Kaplan-Meier method and Cox proportional hazards regression were adopted to estimate CSS, PFS, LPFS, and DMFS. Pearson correlation was used to assess the correlation between variables. We further validated our findings in an independent cohort of 119 patients with ES-NSCLC. RESULTS: A total of 211 patients were included with median follow-up of 48 months in the training cohort. The median EDRIC was 2.178 Gy (range: 0.426-6.015). GTV showed a positive correlation with EDRIC (r = 0.707, P = 0.000). In multivariate analysis, higher EDRIC was significantly associated with worse CSS (HR = 1.468, P = 0.009) and DMFS (HR = 1.491, P = 0.016). Considering each EDRIC quartile, there was a significant difference in CSS between 1st and 4th and 1st and 3rd quartile (P = 0.000, P = 0.004, respectively); and DMFS between 1st and 4th,1st and 3rd, and 1st and 2nd quartile (P = 0.000, P = 0.000, P = 0.008, respectively). In the subgroup and validation cohort, EDRIC was also the important prognostic predictor of CSS and DMFS using multivariate analysis. CONCLUSION: EDRIC was an independent predictor of CSS and DMFS in ES-NSCLC, and it was affected by GTV and tumor location. Though EDRIC is a critical determinant of treatment outcomes, it is quantifiable and potentially modifiable. Additional researches exploring the feasibility of achieving lower EDRIC while maintaining adequate tumor coverage during radiotherapy are warranted.

Antibacterial and immunoregulatory activity of an antimicrobial peptide hepcidin in loach (Misgurnus anguillicaudatus).

Antimicrobial peptides (AMPs) are members of humoral immunity and particpate in resisting microbial invasion. In this study, an AMP gene hepcidin was obtained from the oriental loach Misgurnus anguillicaudatus and named Ma-Hep. This Ma-Hep encodes a peptide of 90 amino acids, with a predicted active peptide segment (Ma-sHep) of 25 amino acids at C terminus. Stimulation by a bacterial pathogen Aeromonas hydrophila resulted in significant up-regulation of Ma-Hep transcripts in loach midgut, head kidney, and gill. Ma-Hep and Ma-sHep proteins were expressed in Pichia pastoris and their antibacterial activity was examined. Results showed that Ma-sHep possessed stronger antibacterial activity against various Gram-positive and Gram-negative bacteria, compared to Ma-Hep. Scanning electron microscopy showed that Ma-sHep might kill bacteria by destroying bacterial cell membranes. Moreover, we found that Ma-sHep had an inhibitory effect on blood cell apoptosis induced by A. hydrophila and facilitated the bacterial phagocytosis and clearance in loach. Histopathological analysis indicated Ma-sHep could protect liver and gut of loach from bacterial infection. Ma-sHep has high thermal stability and PH stability, which is conducive to further feed addition. Feed supplemented with Ma-sHep expressing yeast improved the intestinal flora of loach by increasing the dominant bacteria and decreasing the harmful bacteria. Feed supplemented with Ma-sHep expressing yeast also regulated the expression of inflammatory related factors in various tissues of loach and reduced the mortality of loach upon bacterial infection. These findings show that the antibacterial peptide Ma-sHep is involved in the antibacterial defense of loach and can be used as a candidate for new antimicrobial agents in aquaculture.

[Prognosis Analysis of Early-stage Non-small Cell Lung Cancer Patients Treated with Stereotactic Body Radiotherapy].

BACKGROUND: With the aging of the population and the increased importance of lung cancer screening, the number of early-stage lung cancer patients has been on the rise in recent years, which can be classified into operable early-stage lung cancer and inoperable early-stage lung cancer. The most common pathological type is non-small cell lung cancer (NSCLC). Stereotactic body radiation therapy (SBRT) is the optimal treatment for inoperable early-stage NSCLC. The aim of this study was to investigate the prognosis of early-stage NSCLC patients treated with SBRT and its influencing factors in order to reduce the side effects of radiotherapy and improve the survival and quality of life. METHODS: Clinical data and follow-up outcomes of early-stage NSCLC patients treated with SBRT in our hospital from August 2010 to August 2020 were collected. Kaplan-Meier method was used to assess the prognosis, and the Cox proportional risk model was used for multivariate prognostic analysis. RESULTS: A total of 165 patients were included with a median follow-up time of 43.2 (range: 4.8-132.1) mon. The local control (LC) rates at 1-yr, 2-yr and 5-yr were 98.1%, 94.8% and 86.5% respectively. Karnofsky performance status (KPS) score greater than 80 was an independent prognostic factor for LC (P=0.02). The overall survival (OS) rates at 1-yr, 2-yr and 5-yr were 97.6%, 93.0% and 68.9% respectively. A biological equivalent dose when α/ß=10 (BED10) greater than 132 Gy was an independent prognostic factor for OS (P=0.04). Progression-free survival (PFS) rates at 1-yr, 2-yr and 5-yr were 93.3%, 79.5% and 55.3% respectively. The distance metastasis free survival (DMFS) rates at 1-yr, 2-yr and 5-yr were 94.5%, 83.2% and 58.4% respectively. BED10 greater than 150 Gy was an independent prognostic factor for DMFS (P=0.02). The regional control (RC) rates at 1-yr, 2-yr and 5-yr were 98.8%, 95.4% and 87.9% respectively. CONCLUSIONS: SBRT is effective in treating early-stage NSCLC. KPS greater than 80 is an independent prognostic factor for LC; BED10 greater than 132 Gy is an independent prognostic factor for OS; BED10 greater than 150 Gy is an independent prognostic factor for DMFS.

Impact of aluminum exposure on oxidative stress, intestinal changes and immune responses in red swamp crayfish (Procambarus clarkii).

Aluminum (Al) is an abundant metal that has been classified as a threatening pollutant due to indiscriminate use and anthropogenic activities. This study aimed to evaluate the impacts of Al on crayfish (Procambarus clarkii), including biochemical change, histological alteration, gut microbial community diversification, and immune changes. The bioaccumulation of Al was detected in the hemolymph and intestine of crayfish after Al exposure at different time points. Results showed that Al exposure significantly induced oxidative stress and caused pathohistological changes on intestinal barrier structures in crayfish. It was found that the intestinal microbiota was affected by retained Al and the intestinal community diversity was changed after Al treated in the crayfish. Furthermore, Al exposure affected the immunity in crayfish, by altering the expression of a set of immune-related genes, as well as reducing the phenoloxidase and lysozyme activities. Moreover, Al exposure promoted hemocytes apoptosis and impaired hemophagocytic capacity against Vibro parahamolyticus, resulting in higher mortality of crayfish upon bacterial infection. Taken these results together, we conclude that excessive Al exposure caused adverse effects on multiple biological processes of crayfish and Al pollution is a potential threat to crayfish culture.

Robust prognostic model based on immune infiltration-related genes and clinical information in ovarian cancer.

Immune infiltration of ovarian cancer (OV) is a critical factor in determining patient's prognosis. Using data from TCGA and GTEx database combined with WGCNA and ESTIMATE methods, 46 genes related to OV occurrence and immune infiltration were identified. Lasso and multivariate Cox regression were applied to define a prognostic score (IGCI score) based on 3 immune genes and 3 types of clinical information. The IGCI score has been verified by K-M curves, ROC curves and C-index on test set. In test set, IGCI score (C-index = 0.630) is significantly better than AJCC stage (C-index = 0.541, p < 0.05) and CIN25 (C-index = 0.571, p < 0.05). In addition, we identified key mutations to analyse prognosis of patients and the process related to immunity. Chi-squared tests revealed that 6 mutations are significantly (p < 0.05) related to immune infiltration: BRCA1, ZNF462, VWF, RBAK, RB1 and ADGRV1. According to mutation survival analysis, we found 5 key mutations significantly related to patient prognosis (p < 0.05): CSMD3, FLG2, HMCN1, TOP2A and TRRAP. RB1 and CSMD3 mutations had small p-value (p < 0.1) in both chi-squared tests and survival analysis. The drug sensitivity analysis of key mutation showed when RB1 mutation occurs, the efficacy of six anti-tumour drugs has changed significantly (p < 0.05).

CRISPR-Cas Assisted Shotgun Mutagenesis Method for Evolutionary Genome Engineering.

Genome mutagenesis drives the evolution of organisms. Here, we developed a CRISPR-Cas assisted random mutation (CARM) technique for whole-genome mutagenesis. The method leverages an entirely random gRNA library and SpCas9-NG to randomly damage genomes in a controllable shotgunlike manner that then triggers diverse and abundant mutations via low-fidelity repair. As a proof of principle, CARM was applied to evolve the capacity of Saccharomyces cerevisiae BY4741 to produce ß-carotene. After seven rounds of iterative evolution over two months, a ß-carotene hyperproducing strain, C7-143, was isolated with a 10.5-fold increase in ß-carotene production and 857 diverse genomic mutations that comprised indels, duplications, inversions, and chromosomal rearrangements. Transcriptomic analysis revealed that the expression of 2541 genes of strain C7-143 was significantly altered, suggesting that the metabolic landscape of the strain was deeply reconstructed. In addition, CARM was applied to evolve industrially relevant S. cerevisiae CEN.PK2-1C for S-adenosyl-L-methionine production, which was increased 2.28 times after just one round. Thus, CARM can contribute to increasing genetic diversity to identify new phenotypes that could further be investigated by reverse engineering.

Risk of dietary intake of organochlorine pesticides among the childbearing-age women: A multiple follow-up study in North China.

Exposure to organochlorine pesticides (OCPs) can cause adverse health effects in the female population. We investigated the dietary OCP intake of childbearing-age women living in large agricultural areas of Northern China, as well as their associated health risks. Ten childbearing-age women were recruited during 2015-2016. Their weekly dietary intake diaries and food samples were collected over the course of five visits. The OCP residues of 322 food samples from seven categories (i.e., cereal, vegetable, fruit, fish, meat, egg, and milk) were analyzed by gas chromatography-mass spectrometry. The average concentrations of the total hexachlorocyclohexanes (ΣHCH), dichlorodiphenyltrichloroethanes and their metabolites (ΣDDX), endosulfans (ΣES), and dieldrin and endrin (ΣDrin) in all food categories were, overall, much lower than the maximum residue limits. Relative high mean residues of ΣDrin and ΣES were found in fruits (ΣDrin: 0.687 ng g-1 wet weight (w.w.), ΣES: 2.24 ng g-1 w.w.) and vegetables (ΣDrin: 0.690 ng g-1 w.w., ΣES: 2.11 ng g-1 w.w.). The estimated daily dietary intake (EDI) of these compounds was calculated, with mean levels of 10.6 (ΣES) > 4.37 (ΣDrin) > 1.51 (ΣHCH) > 0.850 (ΣDDX) ng kg-1 day-1. Women during the heating period (from January to March) tended to ingest more ΣHCH, ΣDDX, ΣDrin, and ΣES. Overall, women had no obvious non-carcinogenic and carcinogenic risks due to intake of OCPs, but 83.9% of them has potential carcinogenic risk, with estimated life carcinogenic risk (LCR) exceeding 10-6. Furthermore, women had a higher potential carcinogenic risk during the heating period (mean LCR: 1.33 × 10-5) than during the non-heating period (mean LCR: 8.50 × 10-6). ΣDrin was the dominant OCP responsible for health risks, followed by ΣHCH. We concluded that women in North China still have some dietary OCP intake, especially during the heating period.

Associations of Dietary Exposure to Organochlorine Pesticides from Plant-Origin Foods with Lipid Metabolism and Inflammation in Women: A Multiple Follow-up Study in North China.

This study explored effects of dietary OCP intake from plant-origin foods (cereals, fruits, and vegetables) consumption on lipid metabolism and inflammation of women using a multiple follow-up study. The results showed that dietary intake of p,p'-dichlorodiphenyltrichloroethane (DDT) [ß = - 10.11, 95% confidence interval (95%CI): - 17.32, - 2.905] and o,p'-dichlorodiphenyldichloroethylene (DDE) (ß = - 6.077, 95%CI: - 9.954, - 2.200) were overall negatively associated with serum high-density lipoprotein cholesterol (HDL), whereas other OCPs were not. Serum interleukin (IL)-8 was positively associated with intake of dieldrin (ß = 0.390, 95%CI: 0.105, 0.674), endosulfan-ß (ß = 0.361, 95%CI: 0.198, 0.523), total endosulfan (ß = 0.136, 95%CI: 0.037, 0.234), and total OCPs (ß = 0.084, 95%CI: 0.016, 0.153), and negatively correlated with intake of p,p'-DDE (ß = - 2.692, 95%CI: - 5.185, - 0.198). We concluded that dietary intake of some individual DDT-, DDE- dieldrin-, and endosulfan-class chemicals from plant-origin foods may interfere with lipid metabolism and inflammation responses.

Bioaccessibility dependence of dietary exposure to dichlorodiphenyltrichloroethane and its metabolites and hexachlorocyclohexane isomers and their induced health risk: A case study in Beijing City, China.

Bioaccessibility is essential for evaluating dietary intake of contaminants. However, there is insufficient information on the dependence of dietary intake and risk assessment of dichlorodiphenyltrichloroethane and its metabolites (DDXs) and hexachlorocyclohexane isomers (HCHs) on bioaccessibility. Here, we investigated the bioaccessibilities of DDXs and HCHs in various foods and their influences on assessing exposure in the residents of Beijing City, China. Forty-three major foods in five types (fruit, vegetables, cereals, aquatic food, and meat) were sampled, and the bioaccessibility of DDXs and HCHs was evaluated using a static in vitro gastrointestinal digestion model. The bioaccessibility of DDXs in different food types ranked in the order of meat > vegetables > fruit > cereals > aquatic food, with mean ± standard deviation values of 62.2 ± 22.1%, 20.5 ± 10.6%, 12.4 ± 3.66%, 11.2 ± 9.69%, and 10.7 ± 4.97%, respectively. The highest average bioaccessibility of HCHs was found in meat (83.4 ± 14.2%), followed by fruit (41.0 ± 12.5%), vegetables (37.6 ± 18.1%), aquatic foods (24.2 ± 9.22%), and cereals (8.73 ± 4.07%). The estimated daily intakes (EDI) of the sum of DDXs and the sum of HCHs based on the bioaccessible concentration were only about 17% and 55% of the total EDI based on the residual concentration, respectively. Meat was found to play a more important role in EDI after bioaccessibility correction. The proportion of the population with potential non-carcinogenic and carcinogenic risks markedly decreased when considering bioaccessibility. It was concluded that bioaccessibility should be integrated into dietary exposure evaluation.

Prognostic model of patients with liver cancer based on tumor stem cell content and immune process.

Globally, liver hepatocellular carcinoma (LIHC) has a high mortality and recurrence rate, leading to poor prognosis. The recurrence of LIHC is closely related to two aspects: degree of immune infiltration and content of tumor stem cells. Hence, this study aimed to used RNA-seq and clinical data of LIHC from The Cancer Genome Atlas, Estimation of Stromal and Immune cells in Malignant Tumours, mRNA stemness index score, and weighted gene correlation network analysis methods to find genes significantly linked to the aforementioned two aspects. Key genes and clinical factors were used as input. Lasso regression and multivariate Cox regression were conducted to build an effective prognostic model for patients with liver cancer. Finally, four key genes (KLHL30, PLN, LYVE1, and TIMD4) and four clinical factors (Asian, age, grade, and bilirubin) were included in the prognostic model, namely Immunity and Cancer-stem-cell Related Prognosis (ICRP) score. The ICRP score achieved a great performance in test set. The area under the curve value of the ICRP score in test set for 1, 3, and 5 years was 0.708, 0.723, and 0.765, respectively, which was better than that of other prognostic prediction methods for LIHC. The C-index evaluation method also reached the same conclusion.

A Radiomics Model for Predicting the Response to Bevacizumab in Brain Necrosis after Radiotherapy.

PURPOSE: Bevacizumab is considered a promising therapy for brain necrosis after radiotherapy, while some patients fail to derive benefit or even worsen. Hence, we developed and validated a radiomics model for predicting the response to bevacizumab in patients with brain necrosis after radiotherapy. EXPERIMENTAL DESIGN: A total of 149 patients (with 194 brain lesions; 101, 51, and 42 in the training, internal, and external validation sets, respectively) receiving bevacizumab were enrolled. In total, 1,301 radiomic features were extracted from the pretreatment MRI images of each lesion. In the training set, a radiomics signature was constructed using the least absolute shrinkage and selection operator algorithm. Multivariable logistic regression analysis was then used to develop a radiomics model incorporated in the radiomics signature and independent clinical predictors. The performance of the model was assessed by its discrimination, calibration, and clinical usefulness with internal and external validation. RESULTS: The radiomics signature consisted of 18 selected features and showed good discrimination performance. The model, which integrates the radiomics signature, the interval between radiotherapy and diagnosis of brain necrosis, and the interval between diagnosis of brain necrosis and treatment with bevacizumab, showed favorable calibration and discrimination in the training set (AUC 0.916). These findings were confirmed in the validation sets (AUC 0.912 and 0.827, respectively). Decision curve analysis confirmed the clinical utility of the model. CONCLUSIONS: The presented radiomics model, available as an online calculator, can serve as a user-friendly tool for individualized prediction of the response to bevacizumab in patients with brain necrosis after radiotherapy.

Scalable dextran-polypyrrole nano-assemblies with photothermal/photoacoustic dual capabilities and enhanced biocompatibility.

Polypyrroles have shown great potential in photoacoustic imaging and photothermal therapy owing to its excellent photothermal conversion capabilities. However, the synthesis of polypyrrole-based nano-assemblies which have colloidal stability in biological buffers requires a number of steps, including the polymerization of pyrrole monomers, self-assembly of polypyrrole-based copolymers, and even an additional step to increase the biocompatibility of the nano-assemblies. Herein, a "polymerization/assembly" two-in-one synthesis is proposed for the first time to achieve the one-step synthesis of a new family of polypyrrole-based nano-assemblies, dextran-polypyrrole nano-assemblies (Dex-PPy NAs), under ambient conditions and in aqueous media. In addition, the approach employs tetravalent cerium ions as initiators which can initiate the polymerization of pyrrole monomers through the initiation of free radicals from dextran molecular chains. The resultant Dex-PPy NAs have a photothermal conversion efficiency reaching as high as 41 % and an excellent photostability. More importantly, the NAs with controllable nanoscale dimensions display no signs of cytotoxicity in both in vitro and in vivo studies owing to their biocompatible dextran "shell". An in vivo study further confirmed that the Dex-PPy NAs have excellent real-time photoacoustic imaging and photothermal therapy capabilities for malignant tumors. Therefore, this study represents an important step towards the scalable synthesis of polypyrrole-based nano-assemblies with photothermal/photoacoustic dual capabilities and enhanced biocompatibility.

Identification of Key Genes Related to Lung Squamous Cell Carcinoma Using Bioinformatics Analysis.

Lung squamous cell carcinoma (LUSC) is often diagnosed at the advanced stage with poor prognosis. The mechanisms of its pathogenesis and prognosis require urgent elucidation. This study was performed to screen potential biomarkers related to the occurrence, development and prognosis of LUSC to reveal unknown physiological and pathological processes. Using bioinformatics analysis, the lung squamous cell carcinoma microarray datasets from the Gene Expression Omnibus (GEO) and The Cancer Genome Atlas (TCGA) databases were analyzed to identify differentially expressed genes (DEGs). Furthermore, PPI and WGCNA network analysis were integrated to identify the key genes closely related to the process of LUSC development. In addition, survival analysis was performed to achieve a prognostic model that accomplished good prediction accuracy. Three hundred and thirty-seven up-regulated and 119 down-regulated genes were identified, in which four genes have been found to play vital roles in LUSC development, namely CCNA2, AURKA, AURKB, and FEN1. The prognostic model contained 5 genes, which were all detrimental to prognosis. The AUC of the established prognostic model for predicting the survival of patients at 1, 3, and 5 years was 0.692, 0.722, and 0.651 in the test data, respectively. In conclusion, this study identified several biomarkers of significant interest for additional investigation of the therapies and methods of prognosis of lung squamous cell carcinoma.

Molecular properties, structure, and antioxidant activities of the oligosaccharide Hep-2 isolated from cultured mycelium of Hericium erinaceus.

The oligosaccharide Hep-2 from cultured mycelium of Hericium erinaceus was obtained with a hollow-fiber ultrafiltration cartridge and purified with a DEAE Sephadex A-50 column followed by a Bio-Gel P-30 column. The properties, structure, and antioxidant activities of Hep-2 were studied. Hep-2 had the molecular weight of 1,080 Da and consists of Glc and Gal in a molar ratio of 1.0:0.4. Fragmentation analysis by GC-MS suggests that the structure of Hep-2 consists of three linear sugar residues. Furthermore, we used the LC-MSn combined methylation method to determine the types of bonds between sugar residues. We found that the structure of Hep-2 is based on 2-7 sugars. Among these, the trisaccharides and pentasaccharides consist of 1 â†’ 6-Gal, whereas the tetrasaccharides, hexasaccharides, and heptasaccharides consist of 1 â†’ 4-Glc and 1 â†’ 6-Gal. The activity tests indicated that Hep-2 significantly reduced the damage caused by H2 O2 in GES-1 cells, and could induce expression of T-SOD and GSH-px, scavengers of oxygen free radicals, in a concentration-response manner. Hep-2 also reduced cell apoptosis as assessed by changes in the ratio of Bcl-2/Bax proteins by western blot. Both sets of results suggest that Hep-2 might possess significant antioxidant activity. PRACTICAL APPLICATIONS: This paper reports the physical and the chemical parameters, structure and biological potential of oligosaccharides from Hericium erinaceus, a common edible fungus. Hericium erinaceus has been used as an anti-atrophic gastritis drug in China with good effect. Oligosaccharides are more easily digested and utilized by human body, and have strong antioxidant activity. These results can increase people's interest in the product, and thus have a positive impact on the oligosaccharides of Hericium erinaceus as health food.

PmtA functions as a ferrous iron and cobalt efflux pump in Streptococcus suis.

Transition metals are nutrients essential for life. However, an excess of metals can be toxic to cells, and host-imposed metal toxicity is an important mechanism for controlling bacterial infection. Accordingly, bacteria have evolved metal efflux systems to maintain metal homeostasis. Here, we established that PmtA functions as a ferrous iron [Fe(II)] and cobalt [Co(II)] efflux pump in Streptococcus suis, an emerging zoonotic pathogen responsible for severe infections in both humans and pigs. pmtA expression is induced by Fe(II), Co(II), and nickel [Ni(II)], whereas PmtA protects S. suis against Fe(II) and ferric iron [Fe(III)]-induced bactericidal effect, as well as Co(II) and zinc [Zn(II)]-induced bacteriostatic effect. In the presence of elevated concentrations of Fe(II) and Co(II), ΔpmtA accumulates high levels of intracellular iron and cobalt, respectively. ΔpmtA is also more sensitive to streptonigrin, a Fe(II)-activated antibiotic. Furthermore, growth defects of ΔpmtA under Fe(II) or Co(II) excess conditions can be alleviated by manganese [Mn(II)] supplementation. Finally, PmtA plays a role in tolerance to H2O2-induced oxidative stress, yet is not involved in the virulence of S. suis in mice. Together, these data demonstrate that S. suis PmtA acts as a Fe(II) and Co(II) efflux pump, and contributes to oxidative stress resistance.

An efficient method to simultaneously analyze multi-class organic pollutants in human serum.

The degree of population exposure to various organic pollutants (OPs), including polycyclic aromatic hydrocarbons, organochlorinated pesticides, polychlorinated biphenyls, and polybrominated diphenyl ethers, can be determined by measuring their concentrations in human serum. However, performing large-scale measurements with such a variety of compounds in serum is challenging in terms of efficiency and cost. We describe herein the development of a high-efficiency extraction and sample cleanup protocol for simultaneous and quantitative analyses of OPs using gas chromatography-mass spectrometry. OPs, together with crude lipid impurities, were extracted from human serum with a mixture of n-hexane and methyl tert-butyl ether. A disperse sorbent composed of primary secondary amine and C18 (PSA/C18) was used to roughly remove co-extracted impurities. A combined column of neutral silica gel and neutral alumina oxide (AlO/SiG) was then used for deep cleanup. For the removal of impurities, the overall performance of our protocol for the analysis of OPs in serum was comparable to that of traditional gel permeation chromatography (GPC) and dramatically better than that of PSA/C18, which is a frequently used QuEChERS (quick, easy, cheap, effective, rugged, safe) based method. While both the proposed protocol and GPC yielded recoveries of 80%-110% for four classes of OPs, our protocol consumed about 10 times less solvent, resulting in lower experimental expenses and a lower risk of contamination from residual OPs in the solvent and other supplies. In contrast to GPC, our protocol also permits efficient batch processing of serum samples, allowing for large sample sizes such as those encountered in epidemiological studies.