Dynamic changes of key metabolites during liver fibrosis in rats.

BACKGROUND: Fibrosis is the single most important predictor of significant morbidity and mortality in patients with chronic liver disease. Established non-invasive tests for monitoring fibrosis are lacking, and new biomarkers of liver fibrosis and function are needed. AIM: To depict the process of liver fibrosis and look for novel biomarkers for diagnosis and monitoring fibrosis progression. METHODS: CCl4 was used to establish the rat liver fibrosis model. Liver fibrosis process was measured by liver chemical tests, liver histopathology, and Masson's trichrome staining. The expression levels of two fibrotic markers including α-smooth muscle actin and transforming growth factor ß1 were assessed using immunohistochemistry and real-time polymerase chain reaction. Dynamic changes in metabolic profiles and biomarker concentrations in rat serum during liver fibrosis progression were investigated using ultra-performance liquid chromatography coupled to quadrupole time-of-flight mass spectrometry. The discriminatory capability of potential biomarkers was evaluated by receiver operating characteristic (ROC) curve analysis. RESULTS: To investigate the dynamic changes of metabolites during the process of liver fibrosis, sera from control and fibrosis model rats based on pathological results were analyzed at five different time points. We investigated the association of liver fibrosis with 21 metabolites including hydroxyethyl glycine, L-threonine, indoleacrylic acid, ß-muricholic acid (ß-MCA), cervonoyl ethanolamide (CEA), phosphatidylcholines, and lysophosphatidylcholines. Two metabolites, CEA and ß-MCA, differed significantly in the fibrosis model rats compared to controls (P < 0.05) and showed prognostic value for fibrosis. ROC curve analyses performed to calculate the area under the curve (AUC) revealed that CEA and ß-MCA differed significantly in the fibrosis group compared to controls with AUC values exceeding 0.8, and can clearly differentiate early stage from late stage fibrosis or cirrhosis. CONCLUSION: This study identified two novel biomarkers of fibrosis, CEA and ß-MCA, which were effective for diagnosing fibrosis in an animal model.

Alterations and correlations of the gut microbiome, metabolism and immunity in patients with primary biliary cirrhosis.

We selected 42 early-stage primary biliary cirrhosis (PBC) patients and 30 healthy controls (HC). Metagenomic sequencing of the 16S rRNA gene was used to characterize the fecal microbiome. UPLC-MS/MS assaying of small molecules was used to characterize the metabolomes of the serum, urine and feces. Liquid chip assaying of serum cytokines was used to characterize the immune profiles. The gut of PBC patients were depleted of some potentially beneficial bacteria, such as Acidobacteria, Lachnobacterium sp., Bacteroides eggerthii and Ruminococcus bromii, but were enriched in some bacterial taxa containing opportunistic pathogens, such as γ-Proteobacteria, Enterobacteriaceae, Neisseriaceae, Spirochaetaceae, Veillonella, Streptococcus, Klebsiella, Actinobacillus pleuropneumoniae, Anaeroglobus geminatus, Enterobacter asburiae, Haemophilus parainfluenzae, Megasphaera micronuciformis and Paraprevotella clara. Several altered gut bacterial taxa exhibited potential interactions with PBC through their associations with altered metabolism, immunity and liver function indicators, such as those of Klebsiella with IL-2A and Neisseriaceae with urinary indoleacrylate. Many gut bacteria, such as some members of Bacteroides, were altered in their associations with the immunity and metabolism of PBC patients, although their relative abundances were unchanged. Consequently, the gut microbiome is altered and may be critical for the onset or development of PBC by interacting with metabolism and immunity.

[Research on Real-Time Trace Gas Detection System Based on QEPAS].

Quartz-enhanced photoacoustic spectroscopy (QEPAS) technology was invented lately. Therefore it's an innovative method for trace gas detection compared with other existed technologies. In this paper, we studied the trace gas detection system based on QEPAS, and the atmospheric H2O was selected as the target analyte. In theory, the principles of laser wavelength modulation and signal harmonic detection were analyzed firstly, and the realizing solutions for the gas concentration retrieving and laser wavelength locking were obtained. Furthermore, the selection principle of absorption line for high sensitivity gas detection was discussed. In experiments, a continuous-wave distributed feedback(DFB) single mode diode laser emitting at 1.39 µm was used as the exciting source for the H2O vapor measurement. Using wavelength modulation spectroscopy and 2nd harmonic detection, the influence of laser wavelength modulation depth on QEPAS signal level was investigated, and the acoustic wave enhancement of the addition of micro-resonator in the acoustic detection module was analyzed as well. After optimization of the QEPAS system, a detection limit of 5.9 ppm for H2O vapor was obtained. We measured the H2O vapor with different concentrations, and the R-Square of 0.98 was achieved after the experimental data was linear fitted, indicated that the QEPAS system had an excellent linear response ability. Finally, continuous monitoring of atmospheric H2O concentration levels for a period of 12 hours was performed when the line locking mode was employed with the help of 3rd harmonic detection. The experimental results showed that this QEPAS scheme had a stable performance and outstanding continuous measuring capacity, and it can be widely used in high sensitivity on-line measurement for other trace gases detection fields.

A serum metabonomic study on the difference between alcohol- and HBV-induced liver cirrhosis by ultraperformance liquid chromatography coupled to mass spectrometry plus quadrupole time-of-flight mass spectrometry.

BACKGROUND: Liver cirrhosis is the fatal consequence of chronic hepatitis, making early diagnosis of liver cirrhosis critical. Liver biopsy is still the standard diagnostic method for liver cirrhosis, although its use in a broad population with alcoholism or hepatitis B virus (HBV) infection remains difficult. In this study, we used a metabonomic approach to detect potential biomarkers for early diagnosis of liver cirrhosis. METHODS: Serum specimens were collected prospectively from normal control subjects (n = 22) and patients with alcoholic cirrhosis (n = 18) or HBV-induced cirrhosis (n = 19). The serum metabonome was analyzed using ultraperformance liquid chromatography (LC)/time-of-flight mass spectrometry (MS) integrated with chemometrics. The acquired LC-MS data were normalized and processed using principal component analysis (PCA) and partial least squares discrimination analysis (PLS-DA). RESULTS: Significant differences in the metabonomics among the three groups were observed. Lysophosphatidyl cholines (LPCs) (LPC C16:0, LPC C18:0, LPC C18:2, LPC C18:3, LPC C20:3, LPC C20:5) were decreased in the serum of patients with hepatic cirrhosis, whereas bile acids (glycocholic acid, glycochenodeoxycholic acid), hypoxanthine, and stearamide were increased in the serum of patients with hepatic cirrhosis. These metabolites are considered "common" biomarkers for hepatic cirrhosis. Oleamide and myristamide were increased in the serum of patients with alcoholic cirrhosis but decreased in those with HBV-induced cirrhosis. These could be specific biomarkers for differential diagnosis between alcohol- and HBV-induced hepatic cirrhosis. CONCLUSIONS: There are significant metabonomic differences between alcohol- and HBV-induced liver cirrhosis. Metabonomics is a top-down systems biology tool for conducting research on clinical problems.

Synthesis and biological evaluation of new flavonoid fatty acid esters with anti-adipogenic and enhancing glucose consumption activities.

Oleoyl Formononetin (OF) has good weight loss activity and hypolipidemic activity, could improve insulin sensitivity and suppress adipogenesis. To acquire better biological activities, three series of flavonoid fatty acid esters were designed and synthesized by optimizing the structure of OF. Their bioactivities were assayed in vitro. Some of these novel compounds could effectively inhibit preadipocyte proliferation and adipogenesis. Moreover, they could enhance glucose consumption in adipocytes notably.

[Study of the effect of light source stability on the signal to noise ratio in degenerate four wave mixing experiment].

The effects of the stability of dye laser on the signal to noise ratio in degenerate four-wave mixing (DFWM) were first investigated in iodine vapor using forward geometries. Frequency-doubled outputs from a multi-mode Nd : YAG laser pumped dye laser with laser dye PM580 dissolved in ethanol was used. With the help of forward compensated beam-split technique and imaging detecting system, the saturation intensity of DFWM spectrum in the iodine vapor at 5 554.013 nm was first measured to be 290 microJ under the condition of atmospheric pressure and room temperature. The features of the dye laser such as wavelength ranges, beam quality and energy conversion efficiency decreased gradually with increasing pumping service use, pulse number and intensity. Additionally, with the comparison of the stable and unstable dye laser output, it was found that the instability of dye laser output had greatly influenced the DFWM signal and decreased the signal to background noise ratio. Shot to shot jitter and the broadening in the output frequency leads to an effective broadening of the recorded spectrum and loss of the DFWM signal to noise ratio under the same pumping intensity at different time. The study is of importance to the detection of trace atom, molecule and radical in combustion diagnosis.

[The expression of lefty protein in adult normal skin, human embryonic skin and hyperplastic scar].

OBJECTIVE: To observe the expression of lefty in adult normal skin (ANS), human embryonic skin (HES) and hyperplastic scar (HS), and to explore the effect of lefty on HS and the relationship between lefty and scarless wound healing in embryo. METHODS: Samples of ANS, HES and HS were collected for frozen section for immunofluorescence staining. The morphology of fibroblast and the expression of the lefty were observed by laser confocal microscopy, and the positive cell rates were calculated. RESULTS: Fibroblasts in ANS and HS were long and fusiform with regularity, their nuclei were fusiform or stellate and irregular. Fibroblasts in HES were fusiform, while nuclei were elliptic or fusiform and regular. Positive cell rates of lefty protein in HS (15.38%) were lower than that in NS (67.92%) and FS (81.67%, P < 0.01), and it was lower in ANS compared with HES (P <0.05). CONCLUSION: Lefty protein may inhibit the formation of scar, its high expression may be related to the embryo scarless wound healing.