High-Molecular-Weight Exopolysaccharides Production from Tuber brochii Cultivated by Submerged Fermentation.

Truffles are known worldwide for their peculiar taste, aroma, and nutritious properties, which increase their economic value. However, due to the challenges associated with the natural cultivation of truffles, including cost and time, submerged fermentation has turned out to be a potential alternative. Therefore, in the current study, the cultivation of Tuber borchii in submerged fermentation was executed to enhance the production of mycelial biomass, exopolysaccharides (EPSs), and intracellular polysaccharides (IPSs). The mycelial growth and EPS and IPS production was greatly impacted by the choice and concentration of the screened carbon and nitrogen sources. The results showed that sucrose (80 g/L) and yeast extract (20 g/L) yielded maximum mycelial biomass (5.38 ± 0.01 g/L), EPS (0.70 ± 0.02 g/L), and IPS (1.76 ± 0.01 g/L). The time course analysis of truffle growth revealed that the highest growth and EPS and IPS production was observed on the 28th day of the submerged fermentation. Molecular weight analysis performed by the gel permeation chromatography method revealed a high proportion of high-molecular-weight EPS when 20 g/L yeast extract was used as media and the NaOH extraction step was carried out. Moreover, structural analysis of the EPS using Fourier-transform infrared spectroscopy (FTIR) confirmed that the EPS was ß-(1-3)-glucan, which is known for its biomedical properties, including anti-cancer and anti-microbial activities. To the best of our knowledge, this study represents the first FTIR analysis for the structural characterization of ß-(1-3)-glucan (EPS) produced from Tuber borchii grown in submerged fermentation.

The Diagnostic Value of Artificial Intelligence Ultrasound S-Detect Technology for Thyroid Nodules.

This study aimed to evaluate the consistency of ultrasound TI-RADS classification used by sonographers with different ultrasound diagnosis experience in the diagnosis of thyroid nodules and the diagnostic value of using artificial intelligence ultrasound S-Detect technology in the differentiation of benign and malignant thyroid lesions. 100 patients who underwent ultrasound examination of thyroid masses in our hospital from June 2019 to June 2021 and were further punctured or operated on were included in the study. Pathological results were used as the gold standard to evaluate ultrasound S-Detect technology and the value of TI-RADS classification and the combined application of the two in diagnosing benign and malignant thyroid TI-RADS 4 types of nodules, and the consistency of judgments of doctors of different ages is assessed by a Kappa value. There were 128 nodules in 100 patients, 51 benign nodules, and 77 malignant nodules. For senior physicians, the sensitivity of diagnosis using TI-RADS classification combined with ultrasound S-Detect technology is 93.5%, specificity is 94.1%, and accuracy is 93.8%; for middle-aged physicians using TI-RADS classification combined with ultrasound S-Detect technology for diagnosis, the sensitivity is 89.6%, specificity is 92.2%, and accuracy is 90.6%; for junior doctors, the sensitivity of diagnosis using TI-RADS classification combined with ultrasound S-Detect technology is 83.1%, specificity is 88.2%, and accuracy is 85.1%. Regardless of seniority, the combined application of artificial intelligence ultrasound S-Detect technology and TI-RADS classification can improve the diagnostic ability of sonographers for thyroid nodules and at the same time improve the consistency of judgment among physicians, and this is especially important for radiologists.

Effectiveness of Ultrasound Imaging in Assessing the Palpation Skills of Rotating Physicians.

As an important means of physical examination, palpation is usually limited to the physical examination before surgery and used as an auxiliary method for disease diagnosis in the field of surgery. In practice, palpation is also used in every aspect of the surgical procedure, and its application is of great significance to surgery. The purpose of this study was to investigate the ability of ultrasound imaging to assess the ability of rotating physicians to locate musculoskeletal structures by palpation. Rotating physicians were asked to palpate and locate the long head tendon of the biceps (LHB), posterior tibialis (TPT), acromioclavicular joint (ACJ), and medial tibiofemoral joint (TFJ) spaces on two volunteer models. After positioning, a truncated steel needle was attached to the skin and parallel to the palpable structure, and the position of the steel needle relative to the designated structure was assessed by ultrasound imaging, using the Cohen kappa test to study the inter-rater agreement. The results showed that the assessor's Kappa coefficient for judging the location of all structures was 0.816, LHB was 1.00, TPT was 0.912, ACJ gap was 0.796, and TFJ medial space was 0.844, and the success rate of palpation for TPT was 62.2%, TFJ medial space was 37.8%, ACJ clearance was 24.3%, and LHB was 8.1%. In conclusion, the teaching methods of anatomy and palpation skills need further improvement, and ultrasound imaging is an effective tool for assessing palpation skills.

In vitro activity of tigecycline and proteomic analysis of tigecycline adaptation strategies in clinical Enterococcus faecalis isolates from China.

OBJECTIVES: This study aimed to investigate the in vitro activities of tigecycline (TGC) and the underlying molecular mechanisms of TGC stress response and resistance in clinical Enterococcus faecalis isolates from China. METHODS: Antimicrobial susceptibility and antibiofilm activities of TGC in 399 E. faecalis isolates were evaluated. Heteroresistance was evaluated by population analysis profiling. Resistance and heteroresistance mechanisms were investigated by identifying genetic mutations in tetracycline (tet) target sites and through analysis of efflux protein inhibitors (EPIs). Furthermore, quantitative proteomics was used to investigate the global proteomic response of E. faecalis to TGC stress, as well as the resistance mechanisms of TGC within in vitro induced resistant isolate. RESULTS: TGC minimum inhibitory concentrations (MICs) against clinical E. faecalis isolates were ≤0.5 mg/L. TGC displayed remarkable inhibitory activity against biofilm formation. The occurrence rate of TGC heteroresistance was 1.75% (7/399), and the increased TGC MIC values of heteroresistance-derived clones could be reversed by EPI. TGC resistance was associated with mutations in the 16S rRNA site or 30S ribosomal protein S10. A total of 105 and 356 differentially expressed proteins was identified after being exposed to 1/2× MIC concentrations of TGC, while 356 differentially expressed proteins was identified in TGC-resistant isolate. The differentially expressed proteins were enriched in the translation and DNA replication process. In addition, multiple adenosine triphosphate (ATP)-binding cassette (ABC) transporters were upregulated. CONCLUSIONS: TGC exhibited excellent activity against a substantial proportion of clinical isolates from China. However, E. faecalis exhibited a strong adaptation mechanism during TGC exposure: mutation of TGC target sites and elevated expression of efflux pumps under TGC selection, resulting in TGC resistance.

The Mechanism of Action of Ginkgolic Acid (15:1) against Gram-Positive Bacteria Involves Cross Talk with Iron Homeostasis.

With the increasing reports of community-acquired and nosocomial infection caused by multidrug-resistant Gram-positive pathogens, there is an urgent need to develop new antimicrobial agents with novel antibacterial mechanisms. Here, we investigated the antibacterial activity of the natural product ginkgolic acid (GA) (15:1), derived from Ginkgo biloba, and its potential mode of action against the Gram-positive bacteria Enterococcus faecalis and Staphylococcus aureus. The MIC values of GA (15:1) against clinical E. faecalis and S. aureus isolates from China were ≤4 and ≤8 µg/mL, respectively, from our test results. Moreover, GA (15:1) displayed high efficiency in biofilm formation inhibition and bactericidal activity against E. faecalis and S. aureus. During its inhibition of the planktonic bacteria, the antibacterial activity of GA (15:1) was significantly improved under the condition of abolishing iron homeostasis. When iron homeostasis was abolished, inhibition of planktonic bacteria by GA (15:1) was significantly improved. This phenomenon can be interpreted as showing that iron homeostasis disruption facilitated the disruption of the functions of ribosome and protein synthesis by GA (15:1), resulting in inhibition of bacterial growth and cell death. Genetic mutation of ferric uptake regulator (Fur) led to GA (15:1) tolerance in in vitro-induced resistant derivatives, while overexpression of Fur led to increased GA (15:1) susceptibility. Additionally, GA (15:1) significantly decreased the bacterial loads of S. aureus strain USA300 in the lung tissues of mice in a pneumonic murine model. Conclusively, this study revealed an antimicrobial mechanism of GA (15:1) involving cross talk with iron homeostasis against Gram-positive pathogens. In the future, the natural product GA (15:1) might be applied to combat infections caused by Gram-positive pathogens. IMPORTANCE The increasing emergence of infectious diseases associated with multidrug-resistant Gram-positive pathogens has raised the urgent need to develop novel antibiotics. GA (15:1) is a natural product derived from Ginkgo biloba and possesses a wide range of bioactivities, including antimicrobial activity. However, its antibacterial mechanisms remain unclear. Our current study found that the function of ferric uptake regulator (Fur) was highly correlated with the antimicrobial activity of GA (15:1) against E. faecalis and that the antibacterial activity of GA (15:1) could be strengthened by the disruption of iron homeostasis. This study provided important insight into the mode of action of GA (15:1) against Gram-positive bacteria and suggested that GA (15:1) holds the potential to be an antimicrobial treatment option for infection caused by multidrug-resistant Gram-positive pathogens.

Double contrast-enhanced ultrasonography evaluation of preoperative Lauren classification of advanced gastric carcinoma.

INTRODUCTION: The clinical value of double contrast-enhanced ultrasonography (DCUS) in determining the Lauren classification of advanced gastric carcinoma needed investigation. MATERIAL AND METHODS: Fifty-eight patients with gastric cancer proved by endoscopic biopsy underwent preoperative DCUS examination in which an oral contrast agent was combined with an intravenous agent, and the findings were compared with the postoperative pathological findings using haematoxylin-eosin and Alcian Blue-Periodic Acid Schiff (AB-PAS) staining. RESULTS: Of 58 patients, 34 (59%) were the intestinal type and 24 (41%) the diffuse type on pathological examination of resected specimens. Among intestinal type patients, 30 (88%) showed homogeneous vascular enhancement and 4 (12%) heterogeneous enhancement with the "sandwich" pattern in 2 patients (50%) and "barrier" pattern in 2 patients (50%). In the diffuse type, 22 of 24 patients (92%) enhanced heterogeneously, with stippled and peripheral enhancement in 9 (41%), the "sandwich" pattern in 8 (36%) and "barrier" pattern in 5 (23%). Two of 24 patients (8%) with the diffuse type enhanced homogeneously. The proportion of heterogeneous enhancement was significantly different between the 2 subtypes of tumour (p = 0.0001). The sensitivity and specificity of heterogeneous enhancement in diagnosing the diffuse type of advanced gastric cancer were 92% and 88%, respectively. Youden's index was 0.8. CONCLUSIONS: Double contrast-enhanced ultrasonography is a new and useful method to determine Lauren classification in patients with gastric carcinoma.

Comparative research of the thin transverse sectional anatomy and the multislice spiral CT on Pterygopalatine Fossa.

AIM: To explore a method to obtain sub-millimeter data of the thin transverse section of the pterygopalatine fossa (PPF), and to study the thin transverse sectional anatomy of the adult pterygopalatine fossa and its communicating structure for providing anatomic gist for the imaging diagnosis and minimal invasive operation when PPF diseased. MATERIAL AND METHODS: Two heads of adult cadaver without macroscopic trauma (four sides of PPF) were selected to observe. Images of 0.6 mm-thick multi-planar construction (MPR) were obtained with multislice spiral CT (MSCT) based on the superior orbitomeatal line. Then, the specimens were sliced into 0.1 mm serial section on the transverse plane with the computerized milling machine, the figures were taken with digital camera and the sectional data were stored in the computer. Lastly, the thin transversal section of PPF was investigated and compared with multislice spiral CT images acquired by MPR technique to explore and discuss the anatomy of the thin transverse section of the internal structure of PPF. RESULTS: PPF was divided into four portions: infrapterygopalatine portion, pterygopalatine ganglionic one, suprapterygopalatine one and roof of PPF according to the structural characteristics of the transverse section of PPF. The infrapterygopalatine portion communicated laterally with the infratemporal fossa through the pterygomaxillary fissure and communicated downwards with the oral cavity via palatine greater and lesser canals. The pterygopalatine ganglion was shown clearly in the pterygopalatine ganglionic portion, and its dimensions were 3.91x1.92 mm at the best layer. In the suprapterygopalatine portion, the sphenopalatine foramen and artery were obviously shown on the medial wall, while the palatovaginal canal and artery, the pterygoid canal and artery, and the foramen rotundum and maxillary nerve were shown from the inferiomedial to laterosuperior on the posterior wall. The vomerovaginal canal and artery were located at the slightly superior portion of the medial side of the palatovaginal canal. CONCLUSION: Figures of thin transverse section and multislice spiral CT have highly consistency for the display of PPF. Both of them can correctly identify the micro-structure, the complex relationship of the connectivity and the spatial localization in the narrow space of PPF. It can provide reference gist for the imaging diagnosis and minimal invasive operation.

D-beta-hydroxybutyrate prevents MPP+-induced neurotoxicity in PC12 cells.

Numerous studies show that D-beta-Hydroxybutyrate (DbetaHB) is neuroprotective. The present study was to explore the neuroprotective effects of DbetaHB against the cell death and apoptosis induced by 1-methyl-4-phenylpyridinium ion (MPP+) in PC12 cells. PC12 cells were pretreated with DbetaHB and followed by MPP+ exposure. The cell viability was determined by MTT assay. The morphological characteristics of apoptosis was observed by Acridine Orange (AO) staining and apoptotic rates were measured by flow cytometer. The product of lipid peroxidation, malondialdehyde (MDA), was measured using thiobarbituric acid method. The mitochondrial membrane potential (MMP), intracellular ROS and total glutathione were detected by microplate reader. In PC12 cells, pretreatment with DbetaHB significantly reduced MPP+-induced the decrease of cell viability. AO staining and flow cytometric analysis found DbetaHB inhibited MPP+-induced apoptosis. The measurement of MDA formation showed that DbetaHB alleviated lipid peroxidation induced by MPP+. The loss of MMP induced by MPP+ was preventive by DbetaHB. The changes of intracellular ROS and total glutathione induced by MPP+ were reversed by DbetaHB. DbetaHB protected PC12 cells against MPP+-induced death and apoptosis.