Natural flavonoids: Potential therapeutic strategies for non-alcoholic fatty liver disease.

The incidence of non-alcoholic fatty liver disease (NAFLD) is increasing rapidly worldwide; however, there are currently limited treatments for NAFLD. The disease spectrum includes simple fatty liver, non-alcoholic steatohepatitis (NASH), fibrosis, cirrhosis, and progression to hepatocellular carcinoma (NASH-HCC). The therapeutic effects of NAFLD remain controversial. Although researchers have conducted studies on the pathogenesis of NAFLD, its pathogenesis and anti-NAFLD mechanisms have not been fully elucidated. Previous studies have found that flavonoids, as natural substances with extensive pharmacological activity and good therapeutic effects, have excellent antioxidant, anti-inflammatory, metabolic disease improvement, anti-tumor, and other properties and can significantly alleviate NAFLD. Flavonoids could be further developed as therapeutic drugs for NAFLD. In this paper, the pathogenesis of NAFLD and the mechanisms of flavonoids against NAFLD are summarized to provide a theoretical basis for screening flavonoids against non-alcoholic liver injury.

Untargeted metabolomic approach to study the serum metabolites in women with polycystic ovary syndrome.

BACKGROUND: Polycystic ovary syndrome (PCOS) is not only a kind of common endocrine syndrome but also a metabolic disorder, which harms the reproductive system and the whole body metabolism of the PCOS patients worldwide. In this study, we aimed to investigate the differences in serum metabolic profiles of the patients with PCOS compared to the healthy controls. MATERIAL AND METHODS: 31 PCOS patients and 31 matched healthy female controls were recruited in this study, the clinical characteristics data were recorded, the laboratory biochemical data were detected. Then, we utilized the metabolomics approach by UPLC-HRMS technology to study the serum metabolic changes between PCOS and controls. RESULTS: The metabolomics analysis showed that there were 68 downregulated and 78 upregulated metabolites in PCOS patients serum compared to those in the controls. These metabolites mainly belong to triacylglycerols, glycerophosphocholines, acylcarnitines, diacylglycerols, peptides, amino acids, glycerophosphoethanolamines and fatty acid. Pathway analysis showed that these metabolites were enriched in pathways including glycerophospholipid metabolism, fatty acid degradation, fatty acid biosynthesis, ether lipid metabolism, etc. Diagnosis value assessed by ROC analysis showed that the changed metabolites, including Leu-Ala/Ile-Ala, 3-(4-Hydroxyphenyl) propionic acid, Ile-Val/Leu-Val, Gly-Val/Val-Gly, aspartic acid, DG(34:2)_DG(16:0/18:2), DG(34:1)_DG(16:0/18:1), Phe-Trp, DG(36:1)_DG(18:0/18:1), Leu-Leu/Leu-Ile, had higher AUC values, indicated a significant role in PCOS. CONCLUSION: The present study characterized the difference of serum metabolites and related pathway profiles in PCOS patients, this finding hopes to provide potential metabolic markers for the prognosis and diagnosis of this disease.

DIA proteomics analysis through serum profiles reveals the significant proteins as candidate biomarkers in women with PCOS.

BACKGROUND: The aim of this study was to apply proteomic methodology for the analysis of proteome changes in women with polycystic ovary syndrome (PCOS). MATERIAL AND METHODS: All the participators including 31 PCOS patients and 31 healthy female as controls were recruited, the clinical characteristics data was recorded at the time of recruitment, the laboratory biochemical data was detected. Then, a data-independent acquisition (DIA)-based proteomics method was performed to compare the serum protein changes between PCOS patients and controls. In addition, Western blotting was used to validate the expression of identified proteomic biomarkers. RESULTS: There were 80 proteins differentially expressed between PCOS patients and controls significantly, including 54 downregulated and 26 upregulated proteins. Gene ontology and Kyoto Encyclopedia of Genes and Genomes analysis showed that downregulated proteins were enriched in platelet degranulation, cell adhesion, cell activation, blood coagulation, hemostasis, defense response and inflammatory response terms; upregulated proteins were enriched in cofactor catabolic process, hydrogen peroxide catabolic process, antioxidant activity, cellular oxidant detoxification, cellular detoxification, antibiotic catabolic process and hydrogen peroxide metabolic process. Receiver operating characteristic curves analysis showed that the area under curve of Histone H4 (H4), Histone H2A (H2A), Trem-like transcript 1 protein (TLT-1) were all over than 0.9, indicated promising diagnosis values of these proteins. Western blotting results proved that the detected significant proteins, including H4, H2A, TLT-1, Peroxiredoxin-1, Band 3 anion transport protein were all differently expressed in PCOS and control groups significantly. CONCLUSION: These proteomic biomarkers provided the potentiality to help us understand PCOS better, but future studies comparing systemic expression and exact role of these candidate biomarkers in PCOS are essential for confirmation of this hypothesis.

Pneumocystis jiroveci pneumonia, Nocardia brasiliensis, and Mycobacterium tuberculosis co-infection in a myasthenia gravis patient: A case report.

RATIONALE: Myasthenia gravis (MG) is an autoimmune disorder of the neuromuscular junctions that leads to fluctuating weakness and disabling fatigability. Due to difficulty in breathing caused by weakness of the respiratory muscles, patients with MG are more susceptible to pneumonia and other respiratory infections. As many patients with MG are given immunosuppressive therapy, this makes them more prone to infections. However, coinfection with 3 pathogens is very rare. PATIENT CONCERNS: Here, we report the case of a 41-year-old gentleman with MG who was receiving long-term steroid therapy. He presented with a cough with pale brown expectoration that occurred without obvious inducement, severe pain in the scapula, as well as swelling and weakness of both legs. Despite undergoing treatment, but his symptoms did not improve, prompting two additional hospital admissions over a period of several months. DIAGNOSIS: Bronchoscopy and bronchoalveolar lavage (BAL) were performed, revealing the presence of Pneumocystis jirovecii , Nocardia brasiliensis, and Mycobacterium tuberculosis (MTB). N brasiliensis was identified by positive modified acid-fast Kinyoun staining as well as a positive colony culture identified by matrix-assisted laser desorption ionization-time of flight mass spectrometry from the BAL sample. MTB was confirmed using GeneXpert, and due to the limitations of the culture conditions, methenamine silver stain was used to confirm Pneumocystis jirovecii. Next-generation sequencing (NGS) assay of the BAL samples also confirmed these pathogens. INTERVENTIONS: The patient was transferred to a designated tuberculosis hospital and received anti-infective and anti-TB treatment. OUTCOMES: During treatment at the designated hospital, the patient developed gastrointestinal bleeding and impaired liver function. One month later, he developed multiple organ failure, consolidation of the left lower lung, and pan-drug resistant bacteremia. He refused further treatment and was discharged. CONCLUSION: In conclusion, physicians should be aware of the predisposition of MG patients to co-infections, especially patients with metabolic disorders, to avoid inadequate treatment and poor patient outcomes. Due to the limitations of culture conditions, NGS should be considered as a new technique for identifying pathogens.

Circulating microRNA let­7e is decreased in knee osteoarthritis, accompanied by elevated apoptosis and reduced autophagy.

Knee osteoarthritis (KOA) is a major cause of leg disability in the elderly population. Recently, the expression levels of circulating microRNA (miRNA) let­7e have been reported to be significantly reduced in KOA. The aims of the present study were to assess the feasibility of let­7e as a serum marker for detecting KOA and to explore the underlying mechanisms of its involvement. Based on previous studies and bioinformatics analysis, let­7e may regulate apoptosis and autophagy of articular chondrocytes. A total of 10 patients with KOA and 10 patients with trauma without KOA were recruited to examine the levels of let­7e in peripheral blood. Subsequently, KOA rat models were established, and the levels of let­7e in the cartilage and serum were examined, the expression of apoptotic proteins and autophagy­related proteins in the cartilage were investigated, and apoptotic and autophagic activities of primary cultured chondrocytes were also detected. In patients with KOA, let­7e levels in the peripheral serum were significantly decreased compared with the control group, and this result was confirmed in the peripheral serum and cartilage of KOA rats. In addition, the expression levels of proteins involved in the apoptotic pathway were increased in the cartilage of KOA rats, and apoptotic activity was increased. The expression of autophagy­related proteins beclin 1 and microtubule associated protein 1 light chain 3 ß (LC3B) II/LC3BI in the articular cartilage of KOA rats was lower compared with the controls, and autophagy was decreased. Si­Miao­San (SMS) treatment restored the expression of let­7e and reversed the changes in apoptosis and autophagy. Therefore, the present study provided additional evidence that circulating let­7e may be a potential serum biomarker for the diagnosis and treatment of KOA. Elevated apoptosis levels and decreased autophagy levels of cartilage tissue are involved in KOA, and treatment with SMS may reverse these effects.

[Predominant ß-lactamase genotypes of Escherichia coli isolates and induction and inhibition mechanisms of ß-lactamase gene expression].

OBJECTIVE: To understand the predominant ß-lactamase genotypes and their carrying modes of Escherichia coli isolates in Zhejiang province, and the effects of ß-lactam antibiotics on inducing or histidine kinase inhibitor closantel (CLO) on inhibiting the expression of ß-lactamase genes. METHODS: Micro-dilution method and E-test were applied to measure the resistant rate and minimal inhibitory concentration (MIC) in E. coli isolates against ß-lactam antibiotics. PCR and sequence analysis of PCR products were conducted to detect the ß-lactamase genotypes and their carrying modes. Real-time fluorescent quantitative RT-PCR and ß-lactamase confirmation test were performed to determine the influence of 1/4 MIC penicillin and cefotaxime, and CLO on the transcription and expression of ß-lactamase genes in the resistant E. coli isolates. RESULTS: Among the 462 E. coli strains isolated in Zhejiang, 285 (61.7%) were resistant to penicillin, ampicillin, cefoxitin, cefotaxim and ceftazidime. In the 285 resistant isolates, the detection rate of TEM or CTX-M ß-lactamase gene (83.2% or 75.1%) was significantly higher than that of KPC, SHV or OXA ß-lactamase gene (1.4%-10.2%) (P<0.01) and the carrying rate of two or more ß-lactamase genes (68.8%) was also significantly higher than that of single ß-lactamase gene (31.2%) (P<0.01), and 61.4% of the resistant isolates carried TEM+CTX-M genes (P<0.01). Except KPC gene, 1/4 MIC of cefotaxim and penicillin induced a rapid increase of TEM-mRNA, CTX-M-mRNA, SHV-mRNA or OXA-mRNA levels (P<0.01), but 50-500 µg/ml CLO inhibited these levels (P<0.01). After pre-treatment with 100 µg/ml CLO, 82.8%-85.6% of the resistant isolates became sensitive to ß-lactam antibiotics (P<0.01), while the detection rate of ß-lactamases was also decreased from 95.1% to 16.1% (P<0.01). CONCLUSION: TEM and CTX-M are the predominant ß-lactamase genotypes in E. coli isolates in Zhejiang and TEM+CTX-M is the predominant carrying mode of ß-lactamase genes. Low concentrations of ß-lactam antibiotics can up-regulate the expression levels of ß-lactamase genes in E. coli through bacterial two-component signaling systems, but this effect can be inhibited by CLO, a histidine kinase inhibitor.

[Inhibition effect of hedyotis diffusa wild injection on HL-60 cells and its mechanism].

This study was aimed to explore the inhibition effect and mechanism of hedyotis diffusa wild injection (HDI) on leukemia cell line (HL-60) in vitro. The leukemia cell line HL-60 was used as target cells. The inhibitory effects of HDI on proliferation of HL-60 cells were observed by MTT assay. The positive rate of cell apoptosis and the surface marker of granulocytic differentiation (CD33 and CD15) were measured by flow cytometry. The expressions of anti-apoptosis related gene (survivin and bcl-2) were detected by RT-PCR. The results showed that the growth of HL-60 cells was inhibited by higher concentration of HDI (3.12 - 12.5 ml/L) and inhibited obviously in dose-dependent manner (p < 0.05 or p < 0.01), but not suppressed by low concentration of HDI (1.56 ml/L) in liquid culture system (p > 0.05). The FCM and DNA Ladder results showed that the phenomenon of typical apoptosis did not detected after HL-60 cells were treated with the different concentrations of HDI for 24, 48 and 72 hours respectively. After HL-60 cells were treated with HDI (1.56, 3.12, 6.25 and 12.5 ml/L) for one week, the expression level of CD15 surface marker was all enhanced obviously. When treated with HDI (6.25 ml/L) for 3 weeks, the expression levels of survivin and bcl-2 gene were also decreased obviously by 60% and 44% respectively. It is concluded that HDI can inhibit HL-60 cells in the presence of its higher concentrations. The mechanisms of HDI may induce HL-60 cells differentiation, and suppress the expression of anti-apoptosis related gene (survivin or bcl-2) to inhibit the growth of HL-60 cells.