Comparative analysis of the synergetic effects of Diwuyanggan prescription on high fat diet-induced non-alcoholic fatty liver disease using untargeted metabolomics.

Non-alcoholic fatty liver disease (NAFLD) is one of the most common chronic liver disorders worldwide and had no approved pharmacological treatments. Diwuyanggan prescription (DWYG) is a traditional Chinese medicine preparation composed of 5 kinds of herbs, which has been used for treating chronic liver diseases in clinic. Whereas, the synergistic mechanism of this prescription for anti-NAFLD remains unclear. In this study, we aimed to demonstrate the synergetic effect of DWYG by using the disassembled prescriptions and untargeted metabolomics research strategies. The therapeutic effects of the whole prescription of DWYG and the individual herb were divided into six groups according to the strategy of disassembled prescriptions, including DWYG, Artemisia capillaris Thunb. (AC), Curcuma longa L. (CL), Schisandra chinensis Baill. (SC), Rehmannia glutinosa Libosch. (RG) and Glycyrrhiza uralensis Fisch. (GU) groups. The high fat diets-induced NAFLD mice model was constructed to evaluate the efficacy effects of DWYG. An untargeted metabolomics based on the UPLC-QTOF-MS/MS approach was carried out to make clear the synergetic effect on the regulation of metabolites dissecting the united mechanisms. Experimental results on animals revealed that the anti-NAFLD effect of DWYG prescription was better than the individual herb group in reducing liver lipid deposition and restoring the abnormality of lipidemia. In addition, further metabolomics analysis indicated that 23 differential metabolites associated with the progression of NAFLD were identified and 19 of them could be improved by DWYG. Compared with five single herbs, DWYG showed the most extensive regulatory effects on metabolites and their related pathways, which were related to lipid and amino acid metabolisms. Besides, each individual herb in DWYG was found to show different degrees of regulatory effects on NAFLD and metabolic pathways. SC and CL possessed the highest relationship in the regulation of NAFLD. Altogether, these results provided an insight into the synergetic mechanisms of DWYG from the metabolic perspective, and also supported a scientific basis for the rationality of clinical use of this prescription.

Dynamical changes of tea metabolites fermented by Aspergillus cristatus, Aspergillus neoniger and mixed fungi: A temporal clustering strategy for untargeted metabolomics.

Dark tea fermentation involves various fungi, but studies focusing on the mixed fermentation in tea remain limited. This study investigated the influences of single and mixed fermentation on the dynamical alterations of tea metabolites. The differential metabolites between unfermented and fermented teas were determined using untargeted metabolomics. Dynamical changes in metabolites were explored by temporal clustering analysis. Results indicated that Aspergillus cristatus (AC) at 15 days, Aspergillus neoniger (AN) at 15 days, and mixed fungi (MF) at 15 days had respectively 68, 128 and 135 differential metabolites, compared with unfermentation (UF) at 15 days. Most of metabolites in the AN or MF group showed a down-regulated trend in cluster 1 and 2, whereas most of metabolites in the AC group showed an up-regulated trend in cluster 3 to 6. The three key metabolic pathways mainly composed of flavonoids and lipids included flavone and flavonol biosynthesis, glycerophospholipid metabolism and flavonoid biosynthesis. Based on the dynamical changes and metabolic pathways of the differential metabolites, AN showed a predominant status in MF compared with AC. Together, this study will advance the understanding of dynamic changes in tea fermentation and provide valuable insights into the processing and quality control of dark tea.

Efficacy and safety of a new disposable percutaneous positioning device invented to facilitate the precision of percutaneous core needle lung biopsies: a prospective, open and randomized controlled study.

BACKGROUND: A new disposable percutaneous positioning device was developed which permits adjustment of the fixing puncture angle while performing a percutaneous core needle lung biopsy (CNLB). The aim of the study was to explore the positioning accuracy and clinical safety of the new device during CT-guided percutaneous CNLB. METHODS: A prospective, open and randomized controlled study with two parallel groups was conducted on 150 patients with pulmonary nodules between July 1, 2018 and June 30, 2019 including 101 males and 49 females who were divided (allocation ratio: 1:1) into a standard CNLB group without the percutaneous positioning device (control, n=75) and a new percutaneous positioning device group combined with CNLB (experimental, n=75). The efficacy was determined by the success rate of reaching the target location on the first percutaneous attempt, the number of CT scans performed and the time required for successful puncture positioning. Safety evaluations included complications related to percutaneous surgery and the new positioning device. RESULTS: The success rate of reaching the target location on the first percutaneous attempt in the experimental group was significantly higher than in the control group [96.0% (72/75) vs. 42.7% (32/75), P<0.001]. Patients in the experimental group received 3.05±0.28 times CT scans on average for successful percutaneous positioning, which was significantly lower than for the control group (3.95±1.03) (P<0.001). The time required to complete the percutaneous positioning was significantly lower in the experimental group compared to the control group (8.73±3.11 vs. 12.79±4.55 min, P<0.001). There was no significant difference in percutaneous-related complications between the two groups [48.0% (36/75) vs. 61.3% (46/75), P=0.101], except that the bleeding rate in the experimental group was lower than in the control group [26.7% (20/75) vs. 46.7% (35/75), P=0.032]. CONCLUSIONS: The use of the optimized percutaneous technology with the new positioning device significantly improved the accuracy and precision of percutaneous lung biopsies. TRIAL REGISTRATION: ChiCTR-INR-17010322.

Changes in Triterpenes in Alismatis rhizoma after Processing Based on Targeted Metabolomics Using UHPLC-QTOF-MS/MS.

Alismatis rhizoma (AR) has been used as an herbal medicine in China for over a thousand years. Crude AR, salt-processed AR (SAR), and bran-processed AR (BAR) are recorded in the Pharmacopoeia of the People's Republic of China. However, the differences of chemical composition between crude AR and its processing products remain limited. In this study, triterpenes were identified from crude AR, SAR, and BAR by ultra-high performance liquid chromatography coupled with quadrupole time-of-flight-mass spectrometer (UHPLC-QTOF-MS/MS). Subsequently, the differences of triterpenes between the crude AR and processed ARs were compared via a targeted metabolomics approach. Finally, a total of 114 triterpenes were identified, of which 83, 100, and 103 triterpenes were found in crude AR, SAR, and BAR, respectively. After salt-processing, there were 17 triterpenes newly generated, 7 triterpenes with trends of increasing, and 37 triterpenes decreased. Meanwhile, 56 triterpenes including 21 newly generated and 35 with significant increases were observed in BAR. This study could be benefit to investigate the processing mechanism of AR, as well as support their clinical applications.

Protoporphyrin IX catalyzed hydrogen peroxide to generate singlet oxygen.

AIM: To study the role of protoporphyrin IX (pPIX) in mitochondrial metabolism of hydrogen peroxide (H2O2). METHODS: O2 (-) specific fluorescent markers DMA (9,10-dimerthylanthracence) and SOSG (Singlet Oxygen Sensor Green reagent) were used for measurement of singlet oxygen ((1)O2). Catalyzing conversion of H2O2 into (1)O2 by pPIX was monitored in vitro under varied H2O2 content, temperature, and PH value in the reaction. Ex vivo mitochondrial model was used to analyze effects of ferrochelatase (FECH) and high energy X-rays on this catalytic reaction. RESULTS: In complete dark, measurable (1)O2 was generated when 1.5 mM of H2O2 was incubated with 24 µM of pPIX H2O2 at 37°C for 3 hours. Mitochondrial yield of H2O2 was 0.11±0.03 nmole/mg/min. Mitochondrial FECH significantly improve the catalytic ability of pPIX converting H2O2 into (1)O2. At presence of high-energy X-ray, incubation of 14.4 µM of pPIX with 0.54 µM of H2O2 also generated (1)O2, during which the fluorescence density of 1.05 µM of DMA decreased by 41.5% (P < 0.05). This conversion was not observed when pPIX was replaced with structurally similar hematoporphyrin. CONCLUSION: pPIX can catalyze conversion of H2O2 into (1)O2.