A novel Alisma orientale extract alleviates non-alcoholic steatohepatitis in mice via modulation of PPARα signaling pathway.

Non-alcoholic fatty liver disease (NAFLD), particularly advanced non-alcoholic steatohepatitis (NASH), leads to irreversible liver damage. This study investigated the therapeutic effects and potential mechanism of a novel extract from traditional Chinese medicine Alisma orientale (Sam.) Juzep (AE) on free fatty acid (FFA)-induced HepG2 cell model and high-fat diet (HFD) + carbon tetrachloride (CCl4)-induced mouse model of NASH. C57BL/6 J mice were fed a HFD for 10 weeks. Subsequently, the mice were injected with CCl4 to induce NASH and simultaneously treated with AE at daily doses of 50, 100, and 200 mg/kg for 4 weeks. At the end of the treatment, animals were fasted for 12 h and then sacrificed. Blood samples and liver tissues were collected for analysis. Lipid profiles, oxidative stress, and histopathology were examined. Additionally, a polymerase chain reaction (PCR) array was used to predict the molecular targets and potential mechanisms involved, which were further validated in vivo and in vitro. The results demonstrated that AE reversed liver damage (plasma levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), hepatocyte ballooning, hepatic steatosis, and NAS score), the accumulation of hepatic lipids (TG and TC), and oxidative stress (MDA and GSH). PCR array analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis revealed that AE protects against NASH by regulating the adipocytokine signaling pathway and influencing nuclear receptors such as PPARα. Furthermore, AE increased the expression of peroxisome proliferator-activated receptor gamma coactivator-1α (PPARGC1α) and reversed the decreased expression of PPARα in NASH mice. Moreover, in HepG2 cells, AE reduced FFA-induced lipid accumulation and oxidative stress, which was dependent on PPARα up-regulation. Overall, our findings suggest that AE may serve as a potential therapeutic approach for NASH by inhibiting lipid accumulation and reducing oxidative stress specifically through the PPARα pathway.

A unified deep learning framework for water quality prediction based on time-frequency feature extraction and data feature enhancement.

Deep learning methods exhibited significant advantages in mapping highly nonlinear relationships with acceptable computational speed, and have been widely used to predict water quality. However, various model selection and construction methods resulted in differences in prediction accuracy and performance. Hence, a unified deep learning framework for water quality prediction was established in the paper, including data processing module, feature enhancement module, and data prediction module. In the established model, the data processing module based on wavelet transform method was applied to decomposing complex nonlinear meteorology, hydrology, and water quality data into multiple frequency domain signals for extracting self characteristics of data cyclic and fluctuations. The feature enhancement module based on Informer Encoder was used to enhance feature encoding of time series data in different frequency domains to discover global time dependent features of variables. Finally, the data prediction module based on the stacked bidirectional long and short term memory network (SBiLSTM) method was employed to strengthen the local correlation of feature sequences and predict the water quality. The established model framework was applied in Lijiang River in Guilin, China. The maximum relative errors between the predicted and observed values for dissolved oxygen (DO), chemical oxygen demand (CODMn) were 12.4% and 20.7%, suggesting a satisfactory prediction performance of the established model. The validation results showed that the established model was superior to all other models in terms of prediction accuracy with RMSE values 0.329, 0.121, MAE values 0.217, 0.057, SMAPE values 0.022, 0.063 for DO and CODMn, respectively. Ablation tests confirmed the necessity and rationality of each module for the established model framework. The established method provided a unified deep learning framework for water quality prediction.

Three undescribed coumarins from the roots of Angelica pubescens.

Three undescribed coumarins, 6-(3-hydroxy-3-methyl-2-oxobutyl)-7-methoxycoumarin (1), (R)-6-(1,3-dihydroxy-3-methyl-2-oxobutyl)-7-methoxycoumarin (2), angelol N (6), together with four known coumarins were isolated from the roots of Angelica pubescens. Their structures were determined by extensive spectroscopic analyses. The absolute configurations of compounds 2 and 6 were assigned via electronic circular dichroism (ECD). Their inhibitory effects on lipopoly-saccharide (LPS)-induced nitric oxide (NO) production in RAW264.7 cells were evaluated. Compounds 1-4 exhibited moderate inhibitory activities.

Characteristic of water quality indicators and its response to climate conditions in the middle and lower reaches of Lijiang River, China.

With global climate change and increasingly extreme weather conditions, the water quality of the Lijiang River Basin (LRB) is facing huge threats. At present, there is still a lack of systematic research on water quality indicators and the influence of indirect factors such as meteorological factors on it in the LRB. Therefore, this study is based on the meteorological, hydrological, and water quality data of the LRB from 2012 to 2018, using the Mann-Kendall test, Morlet wavelet method, Spearman's rank correlation coefficient, sensitivity, and contribution rate to quantitative analysis of the relationship between climate conditions and water quality indicators. The results show that the change trends of these hydrological and climatic conditions have almost no significant sudden change; precipitation and streamflow are decreasing each year; the streamflow trend exhibits time hysteresis; precipitation has a stronger influence downstream than on the local area; water quality indicators of both stations exhibited a change period of around 18 to 20 months, with the exception of pH. Water quality indicators are insensitive to precipitation and streamflow, and sensitive to humidity and wind speed; DO was negatively correlated with climate indicators apart from wind speed; almost all water quality indicators in Yangshuo are highly sensitive to air temperature, and the contribution rate of air temperature to ORP and TP reached 4.81% and 3.56%, respectively; sunshine duration has a positive impact on reducing NH4-N and TP. The difference between Yangshuo and Guilin is mostly due to the input of external sources on both sides of the Lijiang River, which results in variations in climate conditions sensitivities.

Comparison of bacterial community structure in PM2.5 during hazy and non-hazy periods in Guilin, South China.

In recent years, significant efforts have been made to study changes in the levels of air pollutants at regional and urban scales, and changes in bioaerosols during air pollution events have attracted increasing attention. In this study, the bacterial structure of PM2.5 was analysed under different environmental conditions during hazy and non-hazy periods in Guilin. A total of 32 PM2.5 samples were collected in December 2020 and July 2021, and the microbial community structures were analysed using high-throughput sequencing methods. The results show that air pollution and climate change alter the species distribution and community diversity of bacteria in PM2.5, particularly Sphingomonas and Pseudomonas. The structure of the bacterial community composition is related to diurnal variation, vertical height, and urban area and their interactions with various environmental factors. This is a comprehensive study that characterises the variability of bacteria associated with PM2.5 in a variety of environments, highlighting the impacts of environmental effects on the atmospheric microbial community. The results will contribute to our understanding of haze trends in China, particularly the relationship between bioaerosol communities and the urban environment. Supplementary Information: The online version contains supplementary material available at 10.1007/s10453-022-09777-0.

Temporal and Spatial Variation Characteristics of Water Quality in the Middle and Lower Reaches of the Lijiang River, China and Their Responses to Environmental Factors.

As the climate and the external environment have changed, the environmental factors of the Lijiang River Basin (LRB) have changed, posing new threats to the environmental quality, ecosystem balance, and management and protection of the water environment of the Lijiang River. Water quality indicators and environmental factors vary spatially along the Lijiang River, which runs through urban areas, farmland, and karst areas. However, research on the response of water quality to water environmental factors is still lacking. Within this context, this study considered statistical methods and hydrological, meteorological, and water quality data of the middle and lower reaches of the Lijiang River from 2012 to 2018, expounded on the temporal and spatial change characteristics and evolution trends of water quality indicators; we analyzed the correlation between water quality indicators and environmental factors; we quantitatively assessed the sensitivity and contribution rate of water quality indicators to environmental factors. The results demonstrated that rainfall feedback on the river streamflow was lagging, and upstream precipitation often affected downstream streamflow. The water quality in the upper reaches of Guilin has improved year by year, and pollution levels have increased slightly when flowing through the urban area of Guilin. In spite of this, it still falls within the range of self-purification. River characteristics heavily influence the impact of environmental factors on water quality indicators; in contrast, the effects of different locations along the same river are more similar. Four water quality indicators are negatively correlated with water temperature, pH, and dissolved oxygen (DO). The sensitivities of ammonia nitrogen (NH4-N) and chemical oxygen demand (CODMn) to streamflow increase with the flow direction. The contribution rates of DO-to-total phosphorus (TP) and pH-to-TP are over -6%. Water temperature is the major contributing factor in the Lijiang River, while DO has a higher contribution in tributaries. The external sources affect the concentration of various water quality indicators and the sensitivity of water quality indicators to the external environment. There should be a series of measures implemented to reduce pollution, such as using oxygenation or chemical means to increase pH in Dahe and Yangshuo to control water pollutants. Tourism and particular karst topography make LRB's calculations unique, but the research method can be applied to other watersheds as well.

Analysis of wet deposition characteristics in the city of Guilin, China.

In this study, the rainfall, pH, conductivity, and ionic component data for Guilin from 2015 to 2017 were analyzed. Specifically, the relationship between the pH value of the rainfall, the change of each ion in the rainfall, and the primary ion sources was examined. The main results obtained were as follows. During the 3-year study period, the average annual pH value of Guilin was 5.45 and exhibited a downward trend. The seasonal variation of rainfall acidity was pronounced, with high pH values and low frequencies of acid rain in summer, and low pH values and high frequencies of acid rain in winter. From 2015 to 2017, the relative order of the average concentrations of the ionic components in the rainfall was SO42- > NO3- > Ca2+ > Cl- > NH4+ > Na+ > K+ > Mg2+ > F-, the annual average concentration of each ionic component displayed a downward trend, and seasonal changes were obvious. Only NH4+ showed an upward trend in rainfall. The (SO42-)/(NO3-) ratio was basically < 3 and manifested a downward trend; (Ca2+)/(NH4+) rose sharply in August and September each year. Using correlation analysis and enrichment factor analysis, it was concluded that the rainfall in Guilin is mainly affected by SO2, NOx, and NH3, with the geological conditions in the karst area also contributing a certain amount to the rainfall acidity. Calculating the enrichment factor revealed that most of the Ca2+ came from a crustal source; half the Mg2+ came from the ocean and half came from the crust; and most of K+ and Cl- originated from the ocean. Human activities contributed most of the SO42- and NO3- in the rainfall.

Four new flavonol glycosides from the leaves of Ginkgo biloba.

Four new flavonol glycosides, 5, 7, 5'-trihydroxy-3', 4'-dimethoxyflavonol-3-O-α-L-rhamnopyranosyl-(1→6)-ß-D-glucopyranoside (1), quercetin 3-O-(6-trans-feruloyl)-ß-D-glucopyranosyl-(1→2)-α-L-rhamnopyranoside (2), kaempferol 3-O-(6-trans-caffeoyl)-ß-D-glucopyranosyl-(1→2)-α-L-rhamnopyranoside (3), myricetin 3-O-(6-trans-p-coumaroyl)-ß-D-glucopyranosyl-(1→2)-α-L-rhamnopyranoside (4), together with nine known flavonoids and two known lignans, were isolated from the leaves of Ginkgo biloba. Their structures were determined by extensive spectroscopic analyses. Their cardioprotective effects against H2O2-induced apoptosis in H9c2 cells were also evaluated. The flavonol glycosides had stronger activity than the acylated flavonol glycosides at the concentration of 50 µM.

[Preparation of baicalein using thermophilic and sugar-tolerant beta-glucosidase].

The reaction conditions of baicalin hydrolyzed into baicalein by a kind of thermophilic and sugar-tolerant beta-glucosidase were studied in this paper. The beta-glucosidase could catalyze baicalin into baicalein well in the acetic acid-sodium acetate buffer. The optimal enzyme activity was at 85 degrees C and pH 5.5. The enzyme was stable at the temperature less than 85 degrees C and pH range of 5-7.5. The maximum reaction rate V. and michaelis constant K. were 0.41 mmol x L(-1) x min(-1) and 3.31 mmol x L(-1) respectively. Different metal ions had different effects on the activity of enzyme. Na+ existing in acetic acid-sodium acetate buffer had an activation effect on enzyme. The enzyme activity was enhanced by the concentrations of glucose below 0.6 mol x L(-1), and was gradually inhibited when monosaccharide concentration was over 0.6 mol x L(-1). When the monosaccharide concentration reached 1.2 mol x L(-1), the inhibition rate of enzyme activity was about 50%, which showed good glucose tolerance. The good reaction conditions through the experiment have been determined as follows, the substrate: enzyme dose was 1 g: 0.2 mL, acetic acid-sodium acetate buffer pH 5.5, reaction temperature 85 degrees C, reaction time 10 h, and the enzymatic hydrolyzation ratio could reach 97%.

Desorption corona beam ionisation (DCBI) mass spectrometry for in-situ analysis of adsorbed phenol in cigarette acetate fiber filter.

The study of spatial distribution characteristics of the adsorbed compounds for absorbent materials has significant importance in understanding the behaviors of aerosols while they migrating in the absorbent materials. Herein, for the first time, desorption corona beam ionization-mass spectrometry (DCBI-MS) has proposed for direct in-situ analysis of adsorbed aerosol for absorbent materials. DCBI is a novel atmospheric pressure chemical ionization (APCI)-related technique developed by our group in recent years. It can facilitate accurately localizing sampling by forming a visible thin corona beam and avoid the risk of sample contamination and matrix interference compared with other similar techniques. The advantages of DCBI-MS allow rapid screening of the spatial distribution characteristics of the adsorbed compounds for absorbent materials. The distribution characteristic of phenol in cigarette filter tip filled with cellulose acetate fiber was studied as a model case for demonstrating the feasibility of the developed method. As a comparison, conventional HPLC was also used for the study of the distribution characteristic of phenol. The results revealed DCBI-MS had highly improved assay simplicity in spatial distribution characteristic analysis of phenol for the acetate fiber tip, therefore, exhibiting a great potential for convenient, rapid and cost-efficient analysis of the spatial distribution characteristic investigation of adsorbed compounds for adsorbent materials.

Adaptive variable-weighted support vector machine as optimized by particle swarm optimization algorithm with application of QSAR studies.

Representing a compound by a numerous structural descriptors becomes common in quantitative structure-activity relationship (QSAR) studies. As every descriptor carries molecular structure information more or less, it seems more advisable to investigate all the possible descriptor vectors rather than traditional variable selection when building a QSAR model. Based on particle swarm optimization (PSO) algorithm, a more flexible descriptor selection and model construction method variable-weighted support vector machine (VW-SVM) is proposed. The new strategy adopted in this paper is to weight all structural descriptors with continuous non-negative values rather than removing or reserving any ones arbitrarily. The manner of invoking PSO to seek non-negative weights of variables can be regarded as a process of searching optimized rescaling for every molecular structural descriptor. Moreover, PSO is employed to search the optimal parameters of VW-SVM model besides variable weights, enables the construction of a rational and adaptive parameter-free QSAR model according to the performance of the total model. Results obtained by investigating glycogen synthase kinase-3α inhibitors and carbonic anhydrase II inhibitors indicate VW-SVM can hold more useful structure information of compounds than other methods as optimally weighting all the descriptors, consequently leading to precisely QSAR models coupled with developed performance both in training and in prediction.

Nucleic acids detection using cationic fluorescent polymer based on one-dimensional microfluidic beads array.

An assay for rapid and direct detection of DNA/mRNA using cationic fluorescent polymer based on one-dimensional microfluidic beads array (1-D chip) has been developed. The cationic water-soluble polythiophene derivatives can easily transduce hybridization events into measurable optical signal due to the conformational changes of the conjugated backbone, when mixed with single-stranded or double-stranded oligonucleotides. In this paper, the polymer was introduced into 1-D chip for fluorescence detection of nucleic acids, and demonstrated its capability on rapid detection of p53 complementary DNA (cDNA) with different concentration. Using this system, we have evaluated the mRNA expression changes of three tumor-associated genes (p53, c-myc and cyclin-d1) in human nasopharyngeal carcinoma CNE2 cell lines before and after 5-flouorouracil (5-FU) stimuli. These results were validated by the conventional reverse transcriptase-PCR. The most important advantage of this assay is not needed target or report labeling prior to hybridization, which makes the experiment process easy to handle and low-cost for multi-target measurement.

Telomerase catalyzed fluorescent probes for sensitive protein profiling based on one-dimensional microfluidic beads array.

A nucleic acid-based signal amplified method for multiple proteins detection based on one-dimensional beads array using telomerase catalyzed fluorescent probes has been developed in this paper. The biotin labeled fluorescent probes were synthesized by telomerase in homogeneous solution. Approximately 360-480 fluorescein molecules were inserted in each probe. The limit of detection for p53 protein is 1.1 pM (S/N=3) and a 3 orders of linear dynamic range is obtained. The sensitivity is nearly two orders higher than the two-site "sandwich" immunoassay using the same platform. Using this method, cellular p53 protein contents of as few as 85 CNE2 cells per mul sample can be determined specifically. The expression changes of p53, c-myc and beta-actin in CNE2 cells were further examined as a function of anti-cancer drug treatment, and the results are consistent with our previous reports. Compared with immuno-polymerase chain reaction and immuno-rolling circle amplification, this method is simple, fast, cheap and suitable for multi-protein analysis. The multiplexed proteins profiling of cellular samples should facilitate the new opportunities to the fundamental research of tumor development and progression, especially to the low abundant tumor-associated proteins analysis.

On-chip oligonucleotide ligation assay using one-dimensional microfluidic beads array for the detection of low-abundant DNA point mutations.

The detection of low-abundant DNA point mutations is very important for the early prediction of cancer, diagnostics of disease and clinical prognosis. In this paper, an on-chip oligonucleotide ligation approach that arrayed a series of functionalized beads in a single microfluidic channel was described for detection of low-abundant point mutations in p53 gene. This gene carried the point mutation with high diagnostic value for assessment of tumor progression and resectional borders. This work extended our prior efforts using one-dimensional (1-D) microfluidic beads array for protein and nucleic acid molecular profiling, and displayed high discrimination sensitivity to mutations detection due to the enhanced mass transport capability caused by microfluidic addressing format of beads array. As a demonstration, it was found that the on-chip beads ligation held high mutation discrimination sensitivity in 1 pM quantities at a SNR (signal-to-noise ratio) >2 using synthesized DNA oligonucleotides in accordance with target fragment. The RT-PCR products of tumor cell line A549, CNE2 and SKBr-3 were further examined to distinguish the point mutation at codon 175 of p53 gene. This approach was capable of detecting a point mutation in a p53 oncogene at a level of 1 mutant in 1000 wild-type sequences using PCR products without the need of LDR amplification. Additionally, this on-chip beads ligation approach also displayed other microfluidic-based advantages of simple handling (one sample injection per test), little reagent quantities, and low potential of contaminations.

Detection of single-base mutations using 1-D microfluidic beads array.

The application of a 1-D microfluidic beads array that is composed of individually addressable functionalized SiO2 beads has been demonstrated for detection of single-base mutations based on "sandwich" hybridization assay without additional sample labeling and PCR amplification. We concentrated on detection of mutations in the human p53 tumor suppressor gene with more than 50% mutation frequency in the known human cancers. Using a microinjection system, functionalized beads could be selectively and linearly arrayed in a single microfluidic channel comprising many periodic chambers. This 1-D microfluidic beads array was sufficiently sensitive to identify single-nucleotide mutations in 40 pM quantities of DNA targets and could discriminate the mutated alleles in an excess of nonmutated alleles at a level of one mutant in 100 wild-type sequences. The surface of beads was regenerated and rehybridized up to six times without obvious loss of signal. The entire reaction process was done at room temperature within minutes, and only 2-10 microL sample solution was needed to complete the whole detection process. The p53 genotypes of A549, CNE2, and SKBr-3 cell lines were also correctly evaluated by using mRNA extracts as target without need for sample labeling and amplification. Thus, this platform enabled rapid and exact discrimination of gene mutations with the advantages of reusability, simple handling of liquid, low cost, and little reagent consumption.

A novel sandwich assay with molecular beacon as report probe for nucleic acids detection on one-dimensional microfluidic beads array.

A novel sandwich assay with molecular beacons as report probes has been developed and integrated into one-dimensional microfluidic beads array (1-D chip) to pursue a label-free and elution-free detection of DNA/mRNA targets. In contrast with the immobilized molecular beacons, this sandwich assay can offer lower fluorescence background and correspondingly higher sensitivity. Furthermore, this sandwich assay on 1-D chip operating in conjunction with molecular beacon technique allows multiple targets detection without the need of laborious and time-consuming elution, which makes the experiment process simple, easy to handle, and reproducible results. In the experiment, the synthesized DNA targets with different concentrations were detected with a detection limit of approximately 0.05 nM. Moreover, the mRNA expression changes in A549 cells before and after anticancer drug 5-flouorouracil treatments were detected and the results were validated by the conventional RT-PCR method.

One-dimensional microfluidic beads array for multiple mRNAs expression detection.

A one-dimensional microfluidic beads array for in vitro rapid measurement of multiple mRNAs expression is presented in this paper. Gene specific capture DNA-functional beads were deposited along a microchannel to form an addressable beads array. We demonstrated that the one-dimensional beads array could perform simultaneous multiple nucleic acid targets detection and a DNA detection limit of 0.02 nM was obtained. Using this array, transcripts expression of three tumor-associated genes, including p53, H-ras, and NME1, both in CNE2 nasopharyngeal carcinoma cell lines and in normal human nasopharyngeal epithelial cells were evaluated. The responses of these three genes expression in CNE2 cells to 5-flouorouracil (5-Fu) stimuli were also assessed. Validation of these results was performed using reverse transcriptase-PCR. The presented methodology combines high throughput of microarrays and low sample consumption, convenient liquid handling of microfluidics. It enables rapid and facile determination of multi-gene expression and holds great potential in early cancer diagnostics and molecular biology.

Quantitative intracellular molecular profiling using a one-dimensional flow system.

We report on the development of one-dimensional microfluidic bead arrays for rapid and quantitative molecular profiling of human cancer cells. This new bioanalytical platform integrates the rapid binding kinetics of suspension bead carriers, the multiplexing and encoding capabilities of gene/protein chips, and the liquid handling advantages of microfluidic devices. Using antibody-conjugated beads in a two-site "sandwich" format, we demonstrate that the proteomic contents of as few as 56 human lung epithelial cancer cells can be determined with high sensitivity and specificity. The results indicate that each cell contains approximately 6 x 10(5) copies of the tumor suppressor protein P53. We have further examined the expression changes of P53, c-Myc, and beta-Actin as a function of anticancer drug treatment and have validated these changes by using Western blotting. This ability to quantitatively analyze normal and diseased cells raises new possibilities in studying cancer heterogeneity and circulating tumor cells.