Discrimination and characterization of volatile organic compounds in Lonicerae Japonicae flos and Lonicerae flos using multivariate statistics combined with headspace gas chromatography-ion mobility spectrometry and headspace solid-phase microextraction gas chromatography-mass spectrometry techniques.

RATIONALE: The volatile organic compounds (VOCs) of Lonicerae Japonicae flos (LJF) and Lonicera flos (LF) play a pivotal role in determining their sensory characteristics, medicinal properties, and subsequent impact on market pricing and consumer preferences. However, the differences and specificity of these VOCs remain obscure. Hence, it is crucial to conduct a comprehensive characterization of the VOCs in LJF and LF and pinpoint their potential differential VOCs. METHODS: In this study, headspace gas chromatography-ion mobility spectrometry (HS-GC/IMS) and headspace solid-phase microextraction gas chromatography-mass spectrometry (HS-SPME-GC/MS) were employed to comprehensively investigate the compositional characteristics and distinctions in VOCs between LJF and LF. Multivariate statistical analysis was used to identify candidate differential VOCs of LJF and LF samples. RESULTS: A total of 54 and 88 VOCs were identified using HS-GC/IMS and HS-SPME-GC/MS analysis, respectively. Primary VOCs detected in LJF include leaf alcohol, (E)-2-hexen-1-ol dimer, 2-octyn-1-ol, and (E)-3-hexen-1-ol. Key VOCs prevalent in LF encompass farnesol, heptanoic acid, octanoic acid, and valeric acid. Multivariate statistical analysis indicates that compounds such as phenethyl alcohol and leaf alcohol were selected as potential VOCs for distinguishing between LJF and LF. CONCLUSION: This research conducted a comprehensive analysis of the fundamental volatile components in both LJF and LF. It subsequently elucidated the distinctions and specificities within their respective VOC profiles. And this study enables differentiation between LJF and LF through the analysis of VOCs, offering valuable insights for enhancing the quality control of both LJF and LF.

Integration of non-targeted multicomponent profiling, targeted characteristic chromatograms and quantitative to accomplish systematic quality evaluation strategy of Huo-Xiang-Zheng-Qi oral liquid.

Huo-Xiang-Zheng-Qi oral liquid (HXZQOL) is a well-known traditional Chinese medicine formula for the treatment of gastrointestinal diseases, with the pharmacologic effects of antiinflammatory, immune protection and gastrointestinal motility regulation. More significantly, HXZQOL is recommended for the treatment of COVID-19 patients with gastrointestinal symptoms, and it has been clinically proven to reduce the inflammatory response in patients with COVID-19. However, the effective and overall quality control of HXZQOL is currently limited due to its complex composition, especially the large amount of volatile and non-volatile active components involved. In this study, aimed to fully develop a comprehensive strategy based on non-targeted multicomponent identification, targeted authentication and quantitative analysis for quality evaluation of HXZQOL from different batches. Firstly, the non-targeted high-definition MSE (HDMSE) approach is established based on UHPLC/IM-QTOF-MS, utilized for multicomponent comprehensive characterization of HXZQOL. Combined with in house library-driven automated peak annotation and comparison of 47 reference compounds, 195 components were initially identified. In addition, HS-SPME-GC-MS was employed to analyze the volatile organic compounds (VOCs) in HXZQOL, and a total of 61 components were identified by comparison to the NIST database, reference compounds as well as retention indices. Secondly, based on the selective ion monitoring (SIM) of 24 "identity markers" (involving each herbal medicine), characteristic chromatograms (CCs) were established on LC-MS and GC-MS respectively, to authenticate 15 batches of HXZQOL samples. The targeted-SIM CCs showed that all marker compounds in 15 batches of samples could be accurately monitored, which could indicate preparations authenticity. Finally, a parallel reaction monitoring (PRM) method was established and validated to quantify the nine compounds in 15 batches of HXZQOL. Conclusively, this study first reports chemical-material basis, SIM CCs and quality evaluation of HXZQOL, which is of great implication to quality control and ensuring the authenticity of the preparation.

The Discrimination and Characterization of Volatile Organic Compounds in Different Areas of Zanthoxylum bungeanum Pericarps and Leaves by HS-GC-IMS and HS-SPME-GC-MS.

The pericarps of Zanthoxylum bungeanum (ZBP) and leaves of Zanthoxylum bungeanum (ZBL) are popular spices in China, and they have pharmacological activities as well. In this experiment, the volatile organic compounds (VOCs) of the pericarps of Zanthoxylum bungeanum in Sichuan (SJ) and its leaves (SJY) and the pericarps of Zanthoxylum bungeanum in Shaanxi (SHJ) and its leaves (SHJY) were analyzed by headspace-gas chromatography-ion mobility spectrometry (HS-GC-IMS) and headspace solid phase microextraction-gas chromatography-mass spectrometry (HS-SPME-GC-MS). The fingerprint of HS-GC-IMS and the heat maps of HS-SPME-GC-MS were established. Principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA) were performed. The results showed that a total of 95 components were identified, 62 components identified by HS-SPME-GC-MS and 40 components identified by HS-GC-IMS, of which 7 were the same. The analysis found that SJ and SHJ were obviously distinguished, while SJY and SHJY were not. There were considerably fewer VOCs in the leaves than in the pericarps. In the characterization of the VOCs of ZBL and ZBP, the flavor of ZBP was more acrid and stronger, while the flavor of ZBL was lighter and slightly acrid. Thirteen and eleven differential markers were identified by HS-GC-IMS and HS-SPME-GC-MS, respectively. This is helpful in distinguishing between SHJ and SJ, which contributes to their quality evaluation.

Discrimination and Characterization of the Volatile Organic Compounds in Schizonepetae Spica from Six Regions of China Using HS-GC-IMS and HS-SPME-GC-MS.

Volatile organic compounds (VOCs) are the main chemical components of Schizonepetae Spica (SS), which have positive effects on the quality evaluation of SS. In this study, HS-SPME-GC-MS (headspace solid-phase microextraction-gas chromatography-mass spectrometry) and HS-GC-IMS (headspace-gas chromatography-ion mobility spectrometry) were performed to characterize the VOCs of SS from six different regions. A total of 82 VOCs were identified. In addition, this work compared the suitability of two instruments to distinguish SS from different habitats. The regional classification using orthogonal partial least squares discriminant analysis (OPLS-DA) shows that the HS-GC-IMS method can classify samples better than the HS-SPME-GC-MS. This study provided a reference method for identification of the SS from different origins.

Ultrahigh-performance liquid chromatography coupled to ion mobility quadrupole time-of-flight mass spectrometry profiling and unveiling the transformation of ginsenosides by the dual conditions of citric acid and high-pressure steaming.

RATIONALE: Many methods have been reported for the production of rare ginsenosides, including heat treatment, acid hydrolysis, alkaline hydrolysis, enzymatic hydrolysis, and microbial transformation. However, the conversion of original ginsenosides to rare ginsenosides under the dual conditions of citric acid and high-pressure steam sterilization has rarely been reported. METHODS: In this study, a method involving ultrahigh-performance liquid chromatography coupled to ion mobility quadrupole time-of-flight mass spectrometry was developed for analysis of chemical transformation of protopanaxatriol (PPT)-type ginsenosides Rg1 and Re, protopanaxadiol (PPD)-type ginsenoside Rb1 , and total ginsenosides in the dual conditions of citric acid and high-pressure steam sterilization. An internal ginsenoside database containing 126 known ginsenosides and 18 ginsenoside reference compounds was established to identify the transformation products and explore possible transformation pathways and mechanisms. RESULTS: A total of 54 ginsenosides have been preliminarily identified in the transformation products of PPD-type ginsenosides Rg1 and Re, PPD-type ginsenoside Rb1 , and total ginsenosides, and the possible transformation pathways were as follows: Rg1 , Re → 20(S)-Rh12 , 20(R)-Rh12 ; Rg1 , Re → 20(S)-Rh1 , 20(R)-Rh1 → Rk3 , Rh4 , Rh5 ; Rb1 → gypenoside LXXV; Rb1 → 20(S)-Rg3 , 20(R)-Rg3 → Rk1 , Rg5 ; Re → 20(S)-Rg2 , 20(R)-Rg2 → 20(S)-Rf2 , 20(R)-Rf2 , Rg4 , F4 . CONCLUSIONS: The results elucidated the possible transformation pathways and mechanisms of ginsenosides in the dual conditions of citric acid and high-pressure steam sterilization, which were helpful for revealing the mechanisms of ginsenosides and enhanced safety and quality control of pharmaceuticals and nutraceuticals. Meanwhile, a simple, efficient, and practical method was developed for the production of rare ginsenosides, which has the potential to produce diverse rare ginsenosides on an industrial scale.

Characterization of γ-Cadinene Enzymes in Ganoderma lucidum and Ganoderma sinensis from Basidiomycetes Provides Insight into the Identification of Terpenoid Synthases.

Enzymes boost protein engineering, directed evolution, and the biochemical industry and are also the cornerstone of metabolic engineering. Basidiomycetes are known to produce a large variety of terpenoids with unique structures. However, basidiomycetous terpene synthases remain largely untapped. Therefore, we provide a modeling method to obtain specific terpene synthases. Aided by bioinformatics analysis, three γ-cadinene enzymes from Ganoderma lucidum and Ganoderma sinensis were accurately predicted and identified experimentally. Based on the highly conserved amino motifs of the characterized γ-cadinene enzymes, the enzyme was reassembled as model 1. Using this model as a template, 67 homologous sequences of the γ-cadinene enzyme were screened from the National Center for Biotechnology Information (NCBI). According to the 67 sequences, the same gene structure, and similar conserved motifs to model 1, the γ-cadinene enzyme model was further improved by the same construction method and renamed as model 2. The results of bioinformatics analysis show that the conservative regions of models 1 and 2 are highly similar. In addition, five of these sequences were verified, 100% of which were γ-cadinene enzymes. The accuracy of the prediction ability of the γ-cadinene enzyme model was proven. In the same way, we also reanalyzed the identified Δ6-protoilludene enzymes in fungi and (-)-α-bisabolol enzymes in plants, all of which have their own unique conserved motifs. Our research method is expected to be used to study other terpenoid synthases with a similar or the same function in basidiomycetes, ascomycetes, bacteria, and plants and to provide rich enzyme resources.

Effect of Oily Sludge Treatment with Molten Blast Furnace Slag on the Mineral Phase Reconstruction of Water-Quenched Slag Properties.

Blast furnace slag, which is the main by-product of the ironmaking process discharged at 1450 °C, contains high-quality sensible heat, while oily sludge is the main solid waste produced in the process of gas exploration, storage, and transportation. The energy and resource utilization of blast furnace slag is complementary to the environmentally friendly treatment of oily sludge, which has provided a new idea for the multi-factor synergistic cycle and energy transformation of the two wastes. The pyrolysis of the oily sludge with the molten blast furnace slag was conducted in the current paper. Results showed that the oily sludge was rapidly pyrolyzed, and the heavy metal elements in the oily sludge were solidified. The solidification rate of the heavy metals exceeds 90%, except for vanadium. The reconstituted water-quenched blast furnace slag still has good activity, and it will not affect the further use of the slag after pyrolysis (BFS-P).

Minimally Invasive Surgery is Superior to Conventional Craniotomy in Patients with Spontaneous Supratentorial Intracerebral Hemorrhage: A Systematic Review and Meta-Analysis.

BACKGROUND: Outcomes of minimally invasive surgery (MIS) versus conventional craniotomy (CC) for patients with spontaneous supratentorial intracerebral hemorrhage (SICH) have not been compared previously. We reviewed the current evidence regarding the safety and efficacy of MIS compared with CC in patients with SICH. METHODS: We conducted a meta-analysis of studies comparing MIS and CC in patients with computed tomography-confirmed SICH published between January 2000 and April 2018 in MEDLINE, Embase, and the Cochrane Controlled Trials Register based on PRISMA inclusion and exclusion criteria. Binary outcomes comparisons between MIS and CC were described using odds ratios (ORs). RESULTS: Five randomized controlled trials (RCTs) and 9 prospective controlled studies (non-RCTs), involving a total of 2466 patients, met our inclusion criteria. There was a statistically significant difference in mortality rate between MIS and CC (OR, 0.76; 95% confidence interval [CI], 0.60-0.97). MIS was associated with a lower rate of rebleeding (OR, 0.42; 95% CI, 0.28-0.64) and a higher rate of good recovery compared with CC (OR, 2.27; 95% CI, 1.34-3.83). CONCLUSIONS: Patients with SICH may benefit more from MIS than CC. Our study could help clinicians optimize treatment strategies in SICH.

Development of a promising fish model (Oryzias melastigma) for assessing multiple responses to stresses in the marine environment.

With the increasing number of contaminants in the marine environment, various experimental organisms have been "taken into labs" by investigators to find the most suitable environmentally relevant models for toxicity testing. The marine medaka, Oryzias melastigma, has a number of advantages that make it a prime candidate for these tests. Recently, many studies have been conducted on marine medaka, especially in terms of their physiological, biochemical, and molecular responses after exposure to contaminants and other environmental stressors. This review provides a literature survey highlighting the steady increase of ecotoxicological research on marine medaka, summarizes the advantages of using O. melastigma as a tool for toxicological research, and promotes the utilization of this organism in future studies.

Deep sequencing-based transcriptome profiling analysis of Oryzias melastigma exposed to PFOS.

Oryzias melastigma is a newly emerging marine fish model. However, the application of this model has been restricted because of the lack of genomic information. Perfluorooctane sulfonate (PFOS), a persistent organic pollutant, is widely distributed in the estuarine/marine environment. The toxicological effects of PFOS on organisms, especially on marine fish species, remain largely unknown. In this study, the transcriptome of O. melastigma was sequenced using newly developed sequencing technology (Illumina RNA-seq). A total of 145,394 unigenes were obtained with 565 bp of unigene N50. These genes were further enriched in various molecular pathways involved in the toxicological response. We also investigated the transcriptional response of O. melastigma embryos after PFOS exposure from 2 days post fertilization (dpf) to 6 dpf by digital gene expression (DGE) technology. The differentially expressed genes were related to neurobehavioral defects, mitochondrial dysfunction and the metabolism of proteins and fats. A further quantitative RT-PCR study showed the down-regulation of ATP synthase and the up-regulation of uncoupling protein 2 (UCP2), which indicated mitochondrial dysfunction. In all, the transcriptome data represent the most comprehensive expressed gene catalog for O. melastigma and will serve as an important reference for various marine fish that are yet to be sequenced. The transcriptome profiling of O. melastigma embryos after exposure to PFOS are also expected to improve our current understanding of the molecular toxicology of PFOS.

PFOS induced precocious hatching of Oryzias melastigma--from molecular level to individual level.

Perfluorooctane sulfonate (PFOS) showed a risk of causing adverse effects including hatching of animals. However, there are no systematical studies about the effects of PFOS on fish embryo hatching. In this study, a precocious hatching was detected in Oryzias melastigma embryos upon PFOS exposure. The average hatching time was shortened and the hatching rate increased after PFOS exposure. The subsequent survival rate decreased significantly in the larvae hatching from the PFOS-treated embryos compared to the control. The influence occurred in a PFOS concentration-dependent manner. To further elucidate the inner molecular change, full length cDNAs of hatching enzymes HCE and LCE were first cloned in O. melastigma by degenerate RT-PCR and RACE. PFOS elicited a transcriptional response of both HCE and LCE. The expressions of both genes were significantly up-regulated earlier in the PFOS-treated group than in the control group. The activity of hatching enzyme also significantly increased upon exposure. In all, our results showed that exposure to PFOS could induce the hatching enzyme both at transcriptional and enzymatic activity level and further lead to the precocious hatching of O. melastigma embryos, which induced the decrease of the subsequent larvae survival.

PFOS elicits transcriptional responses of the ER, AHR and PPAR pathways in Oryzias melastigma in a stage-specific manner.

Perfluorooctane sulfonate (PFOS) is widely distributed in seawater. However, the effects of PFOS on important receptors involved in the toxicity of POPs have not been fully defined, especially for the embryonic stages of marine fish. In this study, we examined the transcriptional responses and PFOS accumulation in the marine medaka embryos at the early and late developmental stages of 4 and 10 dpf upon PFOS exposure (1, 4, and 16mg/L). PFOS accumulated in the embryos, and the embryonic burdens of PFOS at 10 dpf were markedly higher than those at 4 dpf. Moreover, thirteen genes involved in three important POPs-related receptor pathways, including ER, AHR and PPAR, were cloned and investigated. The mRNA expression levels of ERα and ERγ were not significantly altered, but the estrogenic marker genes were downregulated upon PFOS exposure at 4 dpf. Conversely, ERs and related marker genes all were significantly upregulated at 10 dpf. The expressions of ARNT and cyp1a were both upregulated at 4 dpf, while no obvious changes were detected at 10 dpf. The expressions of cyp19a and cyp19b were regulated by PFOS in a stage-specific manner. PFOS produced different effects on three isoforms of PPAR. PPARα and PPARß were first inhibited at 4 dpf and were induced at 10 dpf. PFOS did not elicit a change in PPARγ expression at either stage. In conclusion, this study showed that PFOS has an estrogenic activity and endocrine-disruptive properties. Meanwhile, PFOS could elicit transcriptional responses on POPs-related pathways in a stage-specific manner.

Perfluorooctane sulfonate impairs the cardiac development of a marine medaka (Oryzias melastigma).

Perfluorooctane sulfonate (PFOS) is a persistent organic contaminant and has been widely detected in the sea water. However, toxic effects of PFOS on cardiac development in marine organisms have not been reported. In the present study, we investigated the toxicity of PFOS on the cardiac development using Oryzias melastigma embryos. The embryos at 2 days post-fertilization (dpf) were continuous exposed to PFOS (1, 4 and 16 mg/L) for various periods, cardiac function and morphology were examined at different developmental stages. The results showed that exposure to 4 and 16 mg/L PFOS resulted in enlarged the sinus venosus (SV)-bulbus arteriosus (BA) distance and altered the heart rate. We further investigated eight heart-development related genes to test the effects of PFOS on molecular level. Seven genes were first cloned in O. melastigma and their temporal expression patterns were assayed. Most of the genes were highly expressed in the 6dpf, which is the critical stage for heart development. Their expression levels upon PFOS exposure were studied. The expressions of GATA4 and NKX2.5 were significantly down-regulated while COX-2, FGF8 and ATPase were significantly up-regulated at 6dpf. Our results showed for the first time that PFOS exposure affected the expression of cardiac development-related genes, development and function of heart in the marine medaka.

Embryonic exposure to low concentration of bisphenol A affects the development of Oryzias melastigma larvae.

The prevalence of bisphenol A (BPA) in the environment has attracted increasing attention because of the toxicity of this manmade pollutant. However, the toxicity related to cardiac development remains largely unknown. In the present paper, we investigated the cardiac toxicity of BPA using marine medaka (Oryzias melastigma) embryos. At 2 days postfertilization (dpf), the embryos were continuously exposed to a low concentration of BPA (200 µg/L) for the whole embryonic stage. Heart rate and sinus venosus (SV)-bulbus arteriosus (BA) distance were measured under microscopy. The mRNA expression levels of genes were quantified by SYBR real-time RT-PCR, and hematoxylin and eosin (H&E) staining was used to examine the histology of fish larvae hearts. Neither the heart rate nor the SV-BA distance of the embryos was affected by BPA exposure. However, the mRNA expression levels of Na(+)-K(+)-ATPase, BMP4, COX-1, FGF8, GATA4, and NKX2.5 were all downregulated at the critical developmental stages (6 and 10 dpf). Interestingly, the mRNA expression levels of COX-2 and LERP were significantly upregulated at 10 dpf. The mRNA expressions of inflammation-related genes (TNFα, IL1ß, SOD, and CCL11) were all significantly upregulated after exposure. Moreover, we found that both the body length and the body width decreased in the larvae after embryonic exposure to BPA. The distributed foci of inflammation were observed in the juveniles after 2 weeks' depuration. Exposure to BPA at embryonic stages could alter the expression of heart development-related genes and inflammation-related genes of O. melastigma. The larvae hatched from exposed embryos showed the foci of inflammation in the heart ventricles and the decrease of the body length and width.