Selection and validation of internal control genes for quantitative real-time RT‒qPCR normalization of Phlebopus portentosus gene expression under different conditions.

Phlebopus portentosus (Berk. and Broome) Boedijn is an attractive edible mushroom and is considered the only bolete for which artificial cultivation in vitro has been achieved. Gene expression analysis has become widely used in research on edible fungi and is important for elucidating the functions of genes involved in complex biological processes. Selecting appropriate reference genes is crucial to ensuring reliable RT‒qPCR gene expression analysis results. In our study, a total of 12 candidate control genes were selected from 25 traditional housekeeping genes based on their expression stability in 9 transcriptomes of 3 developmental stages. These genes were further evaluated using geNorm, NormFinder, and RefFinder under different conditions and developmental stages. The results revealed that MSF1 domain-containing protein (MSF1), synaptobrevin (SYB), mitogen-activated protein kinase genes (MAPK), TATA-binding protein 1 (TBP1), and SPRY domain protein (SPRY) were the most stable reference genes in all sample treatments, while elongation factor 1-alpha (EF1), actin and ubiquitin-conjugating enzyme (UBCE) were the most unstably expressed. The gene SYB was selected based on the transcriptome results and was identified as a novel reference gene in P. portentosus. This is the first detailed study on the identification of reference genes in this fungus and may provide new insights into selecting genes and quantifying gene expression.

LC/MS- and GC/MS-based metabolomic profiling to determine changes in flavor quality and bioactive components of Phlebopus portentosus under low-temperature storage.

Introduction: Low temperature is the most common method used to maintain the freshness of Phlebopus portentosus during long-distance transportation. However, there is no information regarding the nutritional changes that occur in P. portentosus preserved postharvest in low temperature. Methods: In this study, the changes in flavor quality and bioactive components in fruiting bodies stored at 4 °C for different storage periods were determined through LC/MS and GC/MS analyses. Sampling was performed at 0, 3, 5, 7, and 13 days storage. Results and Discussion: Based on the results, the metabolites present in caps and stipes were different at the same period and significantly different after 7 days of storage. A total of 583 and 500 different metabolites were detected in caps and stipes, respectively, and were mainly lipids and lipid-like molecules, organic acids and derivatives, organic oxygen compounds and others. Except for prenol lipids and nucleotides, the expression levels of most metabolites increased with longer storage time. In addition, geosmin was identified as the major contributor to earthy-musty odors, and the level of geosmin was increased when the storage time was short. Conclusion: The variations in these metabolites might cause changes in flavor quality and bioactive components in P. portentosus. Variations in these metabolites were thoroughly analyzed, and the results revealed how storage processes affect the postharvest quality of P. portentosus for the first time.

Does temporary ileostomy via specimen extraction site affect the short outcomes and complications after laparoscopic low anterior resection in rectal cancer patients? A propensity score matching analysis.

PURPOSE: The purpose of the current study was to compare the outcomes of temporary stoma through the specimen extraction site (SSES) and stoma through a new site (SNS) after laparoscopic low anterior resection. METHODS: The rectal cancer patients who underwent laparoscopic low anterior resection plus temporary ileostomy were recruited in a single clinical database from Jun 2013 to Jun 2020. The SSES group and the SNS group were compared using propensity score matching (PSM) analysis. RESULTS: A total of 257 rectal cancer patients were included in this study, there were 162 patients in the SSES group and 95 patients in the SNS group. After 1:1 ratio PSM, there was no difference in baseline information (p > 0.05). The SSES group had smaller intraoperative blood loss (p = 0.016 < 0.05), shorter operation time (p < 0.01) and shorter post-operative hospital stay (p = 0.021 < 0.05) than the SNS group before PSM. However, the SSES group shorter operation time (p = 0.006 < 0.05) than the SNS group after PSM, moreover, there was no significant difference in stoma-related complications (p > 0.05). In the multivariate analysis, longer operation time was an independent factor (p = 0.019 < 0.05, OR = 1.006, 95% CI = 1.001-1.011) for the stoma-related complications. CONCLUSION: Based on the current evidence, the SSES group had smaller intraoperative blood loss, shorter operation time and shorter post-operative hospital stay before PSM, and shorter operation time after PSM. Therefore, SSES might be superior than SNS after laparoscopic low anterior resection for rectal cancer patients.

Effect of enhanced recovery after surgery on inflammatory bowel disease surgery: A meta-analysis.

BACKGROUND: The purpose of enhanced recovery after surgery (ERAS) was to reduce surgical pressure and accelerate postoperative functional recovery. Although the application of biologics in treating inflammatory bowel disease (IBD) has changed treatment strategies, most patients with IBD still require surgery. AIM: To evaluate the advantage of ERAS in IBD surgery. METHODS: The PubMed, EMBASE and Cochrane Library databases were searched from inception to March 21, 2021 to find eligible studies. The primary outcome was postoperative complications, and the secondary outcomes included operation time, time to first flatus, time to bowel movement, postoperative hospital stay and readmission. The PROSPERO registration ID of this meta-analysis is CRD42021238052. RESULTS: A total of eight studies involving 1939 patients were included in this meta-analysis. There were no differences in baseline information between the ERAS group and the non-ERAS group. After pooling up all of the data, no significant difference was found between the ERAS group and the non-ERAS group in terms of postoperative overall complications [odds ratio = 0.82, 95% confidence interval (CI) = 0.66 to 1.02, P = 0.08]. The ERAS group had a lower prevalence of anastomotic fistula (odds ratio = 0.36, 95%CI = 0.13 to 0.95, P = 0.04), less time to first flatus [mean difference (MD) = -2.03, 95%CI = -3.89 to -0.17, P = 0.03], less time to bowel movement (MD = -1.08, 95%CI = -1.60 to -0.57, P < 0.01) and shorter postoperative hospital stays (MD = -1.99, 95%CI = -3.27 to -0.71, P < 0.01) than the non-ERAS group. CONCLUSION: ERAS was effective for the quicker recovery in IBD surgery and did not lead to increased complications.

Whole-Genome and Transcriptome Sequencing of Phlebopus portentosus Reveals Its Associated Ectomycorrhizal Niche and Conserved Pathways Involved in Fruiting Body Development.

Phlebopus portentosus (Berk. and Broome) Boedijin, a widely consumed mushroom in China and Thailand, is the first species in the order Boletaceae to have been industrially cultivated on a large scale. However, to date, the lignocellulose degradation system and molecular basis of fruiting body development in P. portentosus have remained cryptic. In the present study, genome and transcriptome sequencing of P. portentosus was performed during the mycelium (S), primordium (P), and fruiting body (F) stages. A genome of 32.74 Mb with a 48.92% GC content across 62 scaffolds was obtained. A total of 9,464 putative genes were predicted from the genome, of which the number of genes related to plant cell wall-degrading enzymes was much lower than that of some saprophytic mushrooms with specific ectomycorrhizal niches. Principal component analysis of RNA-Seq data revealed that the gene expression profiles at all three stages were different. The low expression of plant cell wall-degrading genes also confirmed the limited ability to degrade lignocellulose. The expression profiles also revealed that some conserved and specific pathways were enriched in the different developmental stages of P. portentosus. Starch and sucrose metabolic pathways were enriched in the mycelium stage, while DNA replication, the proteasome and MAPK signaling pathways may be associated with maturation. These results provide a new perspective for understanding the key pathways and hub genes involved in P. portentosus development.

Bacterial Profiling and Dynamic Succession Analysis of Phlebopus portentosus Casing Soil Using MiSeq Sequencing.

Phlebopus portentosus (Berk. and Broome) Boedijin is a popular edible mushroom found in China and Thailand. To date, P. portentosus is the only species in the order Boletales that can be successfully cultivated worldwide. The use of a casing layer or casing soil overlaying the substrate is a crucial step in the production of this mushroom. In this study, bacterial profiling and dynamic succession analyses of casing soil during the cultivation of P. portentosus were performed. One hundred and fifty samples were collected, and MiSeq sequencing of the V3-V4 region of the 16S rRNA gene was conducted. After performing a decontamination procedure, only 38 samples were retained, including 6 casing soil-originated samples (OS), 6 casing soil samples (FHCS) and 5 upper substrate samples (FHCU) from the period of complete colonization by mycelia; 6 casing soil samples (PCS) and 5 upper substrate samples (PCU) from the primordium period; and 6 casing soil samples (FCS) and 4 upper substrate samples (FCU) from fruit body period. The results revealed that bacterial diversity increased sharply from the hyphal to the primordium stage and then decreased during harvesting. The non-metric multidimensional scaling (NMDS) ordination and analysis of similarities (ANOSIM) analysis suggested that the community composition during different stages was significantly different in casing soil. The most abundant phyla in all of the samples were Proteobacteria, Chloroflexi, Acidobacteria, Actinobacteria, Saccharibacteria, and Bacteroidetes. Burkholderia was the most abundant genus in all the samples except the OS samples. The relative abundance of Burkholderia in the FHCS samples (55.79%) decreased to 35.14% in the PCS samples and then increased to 45.60% in the FCS samples. The abundances of Acidobacterium, Rhizobium, Acidisphaera, Bradyrhizobium, and Bacillus increased from the FHCS to PCS samples. The linear discriminant analysis (LDA) effect size (LEfSe) suggested that Acidobacterium and Acidisphaera are micromarkers for PCS, whereas Bradyrhizobium, Roseiarcus, and Pseudolabrys were associated with fruit body stages. The network analyses resulted in 23 edges, including 4 negative and 19 positive edges. Extensive mutualistic interactions may occur among casing soil bacteria. Furthermore, these bacteria play important roles in mycelial elongation, primordium formations, and the production of increased yields.

Clinical significance of serum IGFBP-3 in colorectal cancer.

BACKGROUND: IGF-binding protein 3（IGFBP-3）has previously been identified as tumor marker. The present study aimed to investigate the clinical significance of serum IGFBP-3 in colorectal cancer (CRC). METHODS: Serum was collected from 70 CRC patients and 50 healthy volunteer controls. Serum IGFBP-3 and carcinoembryonic antigen (CEA) levels were measured using electrogenerated chemiluminescence immunoassay and compared between groups. Relationships between serum IGFBP-3 level and the clinical characteristics of CRC were also analyzed. A receiver operator characteristic (ROC) curve was plotted to investigate diagnostic efficacy of serum IGFBP-3 and CEA, respectively, for CRC. Data were analyzed using SPSS 13.0. RESULTS: Serum IGFBP-3 levels in CRC were lower than those of controls (4.68 [3.56, 5.77] vs 5.44 [4.77, 6.10] µg/mL, P < 0.05). Furthermore, serum IGFBP-3 levels were higher in early cancer stages (I and II) than those of advanced stages (III and IV) (4.78 [3.92, 5.49] vs 3.77 [2.65, 4.59] µg/mL, P < 0.05). In addition, patients with lymph node metastasis absent had elevated serum IGFBP-3 levels than those of patients with lymph node metastasis present (4.73 [3.92, 5.72] vs 4.11 [2.45, 4.83] µg/mL, P = 0.02). Finally, ROC curve indicated that serum IGFBP-3 had a better diagnostic power for CRC than CEA. When serum IGFBP-3 and carcinoembryonic antigen were used together to detect CRC, the AUC was 0.949, with a sensitivity of 75% and a specificity of 90%. CONCLUSIONS: Serum IGFBP-3 might be a potential biomarker for CRC.

Incidences of diabetes and prediabetes among female adult breast cancer patients after systemic treatment.

Patients with cancer frequently show glucose intolerance. This study is to estimate the status of total diabetes and prediabetes in breast cancer patients after systemic treatment through an oral glucose tolerance test (OGTT) in China. All the 119 breast cancer patients more than 3 months after systemic treatment with surgery and chemotherapy participated in the study. All the patients without the diagnosis of diabetes underwent OGTT, and fasting and 2-h glucose levels were measured to identify undiagnosed diabetes and prediabetes. Previously diagnosed diabetes were determined on the self-report and the medical record. Of the 119 breast cancer patients, with the median age of 50.1 years and the mean age of about 48 years when they were initially diagnosed with breast cancer, which showed the similar characters of China and Asia breast cancer patients, the overall incidences of total diabetes and prediabetes were 21.8 and 43.7 %, respectively. About 80 % of the diabetes were previously undiagnosed. About 80.0 % of the cases of undiagnosed diabetes and prediabetes met the criteria for elevated 2-h plasma glucose levels through OGTT but not the criteria for elevated fasting glucose levels. Our study firstly documents high incidences of previously undiagnosed diabetes and prediabetes in breast cancer patients during follow-up after systemic treatment through OGTT, indicating that greater diabetes screening, especially through OGTT, prevention, and treatment strategies among breast cancer patients, after systemic treatment for these patients is needed.

[AMPK: a novel target controlling inflammation].

The AMP-activated protein kinase (AMPK) is a pivotal serine/threonine kinase participating in the regulation of glucose, lipid as well as protein metabolism and maintenance of energy homeostasis. Recent studies demonstrated that AMPK can also inhibit nuclear factor-κB, suppress the expression of inflammatory genes and attenuate inflammatory injury through phosphorylating its downstream targets including SIRT1, PGC-lα, p53 and FoxO3a. In addition, the widely used antidiabetic metformin also exerts its anti-inflammatory effects through activating AMPK. Therefore, AMPK is emerging as a promising novel target for the development of anti-inflammatory drugs. This review summarized the anti-inflammatory effects of AMPK and the underling molecular mechanisms.