The Integrated Bioinformatic Assay of Genetic Expression Features and Analyses of Traditional Chinese Medicine Specific Constitution Reveal Metabolic Characteristics and Targets in Steatosis of Nonalcoholic Fatty Liver Disease.

Purpose: In this study, our primary aim is to analyze the genetic expression feature and analyze specific Traditional Chinese medicine (TCM) constitution distribution in non-alcoholic fatty liver disease (NAFLD) and reveal the metabolic characteristic of NAFLD. Materials and Methods: For revealing genetic features, we obtained the gene expression data from the Gene Expression Omnibus (GEO) database of the National Center for Biotechnology Information (NCBI). The genetic data on NAFLD were analyzed by identifying differentially expressed genes (DEGs), associated pathways, co-expressed genetic networks, and gene set enrichment function. Concurrently, we assessed specific constitution distributions among local NAFLD patients through established TCM constitution models and determined the independent variable, including specific constitution to the NAFLD via the regression analyses. Results: The analyses on GEO datasets showed that simple steatosis in NAFLD is strongly associated with HOMA-insulin resistance (HOMA-IR). Analyses of GEO datasets revealed significantly altered genetic expression profiles between NAFLD and normal populations. For TCM constitution analyses, we demonstrated a decline in yin-yang harmony (YYH) and yang-asthenia (YAAC) constitution, whereas there was an increase in qi-stagnation (QSC) and phlegm-dampness (PDC) in NAFLD. The binary logistic regression analysis indicated that besides other metabolic parameters, YYH, qi asthenia (QAC), YYAC, and yin-asthenia (YAC) were the independent variables of NAFLD, while YAC was the independent variables of T2D. The multilinear regression analyses suggested that NAFLD, DM, BMI, waist, TC, TG, hypertension, ALT, AST, and YAC were the significant determinators of the FPG. Conclusion: This study presents a relatively comprehensive metabolic profile in steatosis of NAFLD, revealed by significant genetic expression feature alterations and different TCM constitution distribution in NAFLD. Through this method, the study intends to associate the genetic feature with the phenotype of TCM constitution. The results could be applied to assist integrative medicine research in exploring the appropriate personalized approaches for NAFLD.

Whole genome sequencing of multidrug-resistant Proteus mirabilis strain PM1162 recovered from a urinary tract infection in China.

OBJECTIVES: Proteus mirabilis is an important opportunistic Gram-negative pathogen. This study reports the whole genome sequence of multidrug-resistant (MDR) P. mirabilis PM1162 and explores its antibiotic resistance genes (ARGs) and their genetic environments. METHODS: P. mirabilis PM1162 was isolated from a urinary tract infection in China. Antimicrobial susceptibility was determined, and whole genome sequencing (WGS) was performed. ARGs, insertion sequence (IS) elements, and prophages were identified using ResFinder, ISfinder, and PHASTER software, respectively. Sequence comparisons and map generation were performed using BLAST and Easyfig, respectively. RESULTS: On its chromosome, P. mirabilis PM1162 harboured 15 ARGs, including cat, tet(J), blaCTX-M-14 (three copies), aph(3')-Ia, qnrB4, blaDHA-1, qacE, sul1, armA, msr(E), mph(E), aadA1, and dfrA1. We focused our analysis on the four related MDR regions: (1) genetic contexts associated with blaCTX-M-14; (2) the prophage containing blaDHA-1, qnrB4, and aph(3')-Ia; (3) genetic environments associated with mph(E), msr(E), armA, sul, and qacE; and (4) the class II integron harbouring dfrA1, sat2, and aadA1. CONCLUSION: This study reported the whole genome sequence of MDR P. mirabilis PM1162 and the genetic context of its ARGs. This comprehensive genomic analysis of MDR P. mirabilis PM1162 provides a deeper understanding of its MDR mechanism and elucidates the horizontal spread of its ARGs, thus providing a basis for the containment and treatment of the bacteria.

CircLDLR acts as a sponge for miR-667-5p to regulate SIRT1 expression in non-alcoholic fatty liver disease.

BACKGROUND: Non-alcoholic fatty liver (NAFLD) is a complex metabolic disease characterized by fatty degeneration of hepatocytes. Circular RNAs (circRNAs) have been reported to be essential for (NAFLD progression. The potential mechanism of circRNA low-density lipoprotein receptor (circLDLR) in the NAFLD was investigated in this study. METHODS: Hepatocyte (Hepa1-6) cells treated with oleic acid/palmitic acid (OA/PA) were used as the in vitro NAFLD model, and C57BL/6 mice fed with high-fat diet (HFD) were used as the in vivo NAFLD model. The circLDLR, LDLR, and miR-667-5p expression were measured by quantitative real-time polymerase chain reaction (qRT-PCR), while the protein levels of Light Chain Microtubule-Associated Protein 3 (LC3) and Sequestosome-1(p62) was examined by western blot. The circLDLR location was confirmed using RNA fluorescence in situ hybridization. Oil red O staining was carried out to measure lipid deposition in cells. The secreted levels of triglyceride (TG) and total cholesterol (TC) were detected through Enzymatic. The existence of the circLDLR/miR-667-5p/sirtuin 1 (SIRT1) regulatory axis was validated by applying the dual-luciferase reporter assay. RESULTS: The circLDLR expression showed a prominent down-regulation in OA/PA-treated Hepa1-6 cells, whereas the LDLR expression was up-regulated. Overexpression of circLDLR significantly attenuated lipid droplet accumulation in NAFLD models in vitro/vivo, reduced TG, TC, and p62 levels, and increased LC3-II levels and the amount of the green fluorescent protein (GFP)-LC3 puncta in cells. CircLDLR and SIRT1 are common targets of miR-667-5p to inhibit the TG and TC and promote the autophagy pathway. SIRT1 knockdown reversed the effects of circLDLR overexpression. CONCLUSIONS: CircLDLR alleviated the development of NAFLD by inducing autophagic flux while modulating the miR-667-5p/SIRT1 axis reversed its effects, suggesting that targeting circLDLR/miR-667-5p/SIRT1 axis may be a promising therapeutic strategy for NAFLD.

A Pilot Study of Modified Mini-Clinical Evaluation Exercises (Mini-CEX) in Rotation Students in the Department of Endocrinology.

Background: The mini-clinical evaluation exercise (mini-CEX) is an excellent tool for assessing the clinical abilities of medical students in intense clinical practice. In this study, the Mini-CEX was adapted to professional questionnaires for Diabetes Mellitus (DM), and examined in medical students completing their clerkship rotation in the department of endocrinology. Methods: From January 2021 to January 2022, all rotating medical students at Shanghai Pudong Hospital completed two mini-CEX exams before and following their rotation under the supervision and guidance of six tutors. The mini-CEX form was modified in this study primarily for inpatient management based on our clinical experience and updated DM guidelines of the American Diabetes Association (ADA), the European Association for the Study of Diabetes (EASD), and the Chinese Diabetes Society (CDS). Each component of the mini-CEX assessment, including medical interviews, physical examination, clinical judgment, clinical management, and overall clinical competence was evaluated using a nine-item questionnaire. Results: Our findings revealed that the second-round performance on the assessments significantly improved, as indicated by higher scores on each component. The Pearson association analysis revealed that the feedback time of the first examination was markedly associated with improved overall scores (r= 0.391, p<0.001). However, no correlations were discovered between patient age, gender, disease severity disparity, or the interval between examinations (p>0.05). Additional regression analysis revealed that the feedback time during the initial examination was the most significant contributor to the increased overall scores (ß=0.391, p<0.001). Conclusion: This newly designed mini-CEX form based on current ADA and EASD guidelines may assist trainees in more effectively diagnosing and managing DM in inpatients, particularly those with macrovascular, microvascular, or peripheral nerve neuropathy. This study aims to assess the efficacy of administering a modified mini-CEX form to rotating trainees participating in an endocrine clerkship.

Clinical Utility of Circulating Tumor Cells in the Early Detection of Lung Cancer in Patients with a Solitary Pulmonary Nodule.

Objective: Lung cancer is the most common cancer and can appear as a solitary pulmonary nodule. Early detection of lung cancer in this patient population would be beneficial for the disease management. In this study, the potential application of circulating tumor cells (CTCs) on early detection of lung cancer in this population was investigated. Methods: The number of CTCs in bronchoalveolar lavage fluid and serum levels of tumor-related markers, cancer antigen 125 (CA125), carcinoembryonic antigen (CEA) and neuron-specific enolase (NSE) were measured in patients with a solitary pulmonary nodule. The association between CTCs and lung cancer was examined. The diagnosis performances of CTCs and selected tumor-related markers were compared. Results: The CTC positivity was significantly associated with lung cancer (P = .009). The sensitivity of CTCs and CA125, CEA, NSE, and CA125/CEA/NSE was 75%, 5.6%, 0%, 25%, and 33%, respectively. The sensitivity of CTCs was improved from 75% to 83% by the combination with CA125 or NSE. Conclusion: CTCs may be helpful for the early detection of lung cancer in patients with a solitary pulmonary nodule.

[Effect of vertical anterior laryngectomy with pedicled sternohyoid flap for vocal cord reconstruction and glottal morphology].

Objective:To explore the effect of vocal cord reconstruction with sternohyoid muscle flap pedicled with vertical anterior laryngectomy. Method:The clinical data of 43 cases of laryngeal carcinoma were analyzed retrospectively. According to whether the vocal cords were reconstructed, they were divided into vocal cord reconstruction group and non reconstruction group. Among them, 20 patients in the reconstruction group were reconstructed with pedicled sternohyoid muscle flap and 23 patients in non-reconstruction group. In the non reconstruction group, the external membrane of thyroid cartilage was used to repair the wounds. Postoperative respiratory function, swallowing function, pronunciation function, postoperative prognosis, complications, and recurrence rate were compared between the two groups. Reconstruction of glottis after vocal cord reconstruction was evaluated by electronic laryngoscope and CT scan. Result:①Patients in both groups survived during the follow-up period. One patient in the non-reconstructed group had recurrence, and the reconstituted group had no relapse, 3 cases with complications occurred in the reconstruction group, including 2 cases with granulation tissue in the glottic area, 1 case with laryngeal fistula, and 2 cases with aspiration pneumonia were found in the non-reconstruction group. ②1 year postoperative tracheal cannula removal rate, gastric tube removal and pronunciation quality: the tracheal cannula removal rate was 100% in the two groups after surgery; the gastric tube removal time in the reconstruction group was（13.2±2.8） days, and （16.6±5.3） days in the non-reconstruction group （P<0.05）; reconstruction group had good pronunciation in 10 cases, moderate in 6 cases, and poor in 4 cases. Non-reconstructed group had good pronunciation in 4 cases, medium in 14 cases, and poor in 5 cases. Those with moderate or higher were compared no significant difference （P>0.05）, and those with good pronunciation were statistically different （P<0.05）. ③The transverse and anteroposterior diameter in reconstruction group was similar with the normal people （P>0.05）; however, the transverse and anteroposterior diameter in the non-reconstructed group was significantly different with that of the normal people （P>0.05）, the transverse diameter of the reconstructed group and the non-reconstructed group were compared with no significant difference （P>0.05）, but there was significant difference in the anteroposterior diameter between the two groups （P<0.05）. The area in both groups were different with the normal people （P<0.05）; ④Glottic area morphology: the two groups of patients showed different degrees of swelling in the arytenoid cartilage area, the shape of the glottic region in the reconstructed group was approximately triangular, and the glottic morphology in the non-reconstructed group was approximately circular. Conclusion:After vocal cord reconstruction, there were increased rate of tracheal cannula extubation, well covered wallowing and phonation function, and the quality of life of patients was improved.

Astragalus polysaccharides affect insulin resistance by regulating the hepatic SIRT1-PGC-1α/PPARα-FGF21 signaling pathway in male Sprague Dawley rats undergoing catch-up growth.

The present study investigated the effects of Astragalus polysaccharides (APS) on insulin resistance by modulation of hepatic sirtuin 1 (SIRT1)­peroxisome proliferator­activated receptor (PPAR)­Î³ coactivator (PGC)­1α/PPARα­fibroblast growth factor (FGF)21, and glucose and lipid metabolism. Thirty male Sprague Dawley rats were divided into three groups: A normal control group, a catch­up growth group and an APS­treated (APS-G) group. The latter two groups underwent food restriction for 4 weeks, prior to being provided with a high fat diet, which was available ad libitum. The APS­G group was orally treated with APS for 8 weeks, whereas the other groups were administered saline. Body weight was measured and an oral glucose tolerance test (OGTT) was conducted after 8 weeks. The plasma glucose and insulin levels obtained from the OGTT were assayed, and hepatic morphology was observed by light and transmission electron microscopy. In addition, the mRNA expression levels of PGC­1α/PPARα, and the protein expression levels of SIRT1, FGF21 and nuclear factor­κB were quantified in the liver and serum. APS treatment suppressed abnormal glycolipid metabolism and insulin resistance following 8 weeks of catch­up growth by improving hepatic SIRT1­PPARα­FGF21 intracellular signaling and reducing chronic inflammation, and by partially attenuating hepatic steatosis. The suppressive effects of APS on liver acetylation and glycolipid metabolism­associated molecules contributed to the observed suppression of insulin resistance. However, the mechanism underlying the effects of APS on insulin resistance requires further research in order to be elucidated. Rapid and long­term treatment with APS may provide a novel, safe and effective therapeutic strategy for type 2 diabetes.

Exendin-4 Promotes Beta Cell Proliferation via PI3k/Akt Signalling Pathway.

BACKGROUND/AIMS: Prevention of diabetes requires maintenance of a functional beta-cell mass, the postnatal growth of which depends on beta cell proliferation. Past studies have shown evidence of an effect of an incretin analogue, Exendin-4, in promoting beta cell proliferation, whereas the underlying molecular mechanisms are not completely understood. METHODS: Here we studied the effects of Exendin-4 on beta cell proliferation in vitro and in vivo through analysing BrdU-incorporated beta cells. We also analysed the effects of Exendin-4 on beta cell mass in vivo, and on beta cell number in vitro. Then, we applied specific inhibitors of different signalling pathways and analysed their effects on Exendin-4-induced beta cell proliferation. RESULTS: Exendin-4 increased beta cell proliferation in vitro and in vivo, resulting in significant increases in beta cell mass and beta cell number, respectively. Inhibition of PI3K/Akt signalling, but not inhibition of either ERK/MAPK pathway, or JNK pathway, significantly abolished the effects of Exendin-4 in promoting beta cell proliferation. CONCLUSION: Exendin-4 promotes beta cell proliferation via PI3k/Akt signaling pathway.