Comprehensive metabolome characterization and comparison between two sources of Dragon's blood by integrating liquid chromatography/mass spectrometry and chemometrics.

Dragon's Blood (DB) serves as a precious Chinese medicine facilitating blood circulation and stasis dispersion. Daemonorops draco (D. draco; Qi-Lin-Jie) and Dracaena cochinchinensis (D. cochinchinenesis; Long-Xue-Jie) are two reputable plant sources for preparing DB. This work was designed to comprehensively characterize and compare the metabolome differences between D. draco and D. cochinchinenesis, by integrating liquid chromatography/mass spectrometry and untargeted metabolomics analysis. Offline two-dimensional liquid chromatography/ion mobility-quadrupole time-of-flight mass spectrometry (2D-LC/IM-QTOF-MS), by utilizing a powerful hybrid scan approach, was elaborated for multicomponent characterization. Configuration of an XBridge Amide column and an HSS T3 column in offline mode exhibited high orthogonality (A0 0.80) in separating the complex components in DB. Particularly, the hybrid high-definition MSE-high definition data-dependent acquisition (HDMSE-HDDDA) in both positive and negative ion modes was applied for data acquisition. Streamlined intelligent data processing facilitated by the UNIFI™ (Waters) bioinformatics platform and searching against an in-house chemical library (recording 223 known compounds) enabled efficient structural elucidation. We could characterize 285 components, including 143 from D. draco and 174 from D. cochinchinensis. Holistic comparison of the metabolomes among 21 batches of DB samples by the untargeted metabolomics workflows unveiled 43 significantly differential components. Separately, four and three components were considered as the marker compounds for identifying D. draco and D. cochinchinenesis, respectively. Conclusively, the chemical composition and metabolomic differences of two DB resources were investigated by a dimension-enhanced analytical approach, with the results being beneficial to quality control and the differentiated clinical application of DB.

[Variations of glucose content in Massa Medicata Fermentata during processing based on quantitative proton nuclear magnetic resonance].

A quantitative proton nuclear magnetic resonance(qHNMR) method was established to determine the glucose content in commercially available Massa Medicata Fermentata(MMF) products and explore the variations of glucose content in MMF products during processing. The qHNMR spectrum of MMF in deuterium oxide was obtained with 2,2,3,3-d_4-3-(trimethylsilyl) propionate sodium salt as the internal standard substance. With the doublet peaks of terminal hydrogen of glucose with chemical shift at δ 4.65 and δ 5.24 as quantitative peaks, the content of glucose in MMF samples was determined. The glucose content showed a good linear relationship within the range of 0.10-6.44 mg·mL~(-1). The relative standard deviations(RSDs) of precision, stability, repeatability, and recovery for determination were all less than 2.3%. The glucose content varied in different commercially available MMF samples, which were associated with the different fermentation days, wheat bran-to-flour ratios, and processing methods. The glucose content in MMF first increased and then decreased over the fermentation time. Compared with the MMF products fermented with wheat bran or flour alone, the products fermented with both wheat bran and flour had increased glucose. The glucose content of bran-fried MMF was slightly lower than that of raw MMF, while the glucose content in charred MMF was extremely low. In conclusion, the qHNMR method established in this study is simple, fast, and accurate, serving as a new method for determining the glucose content in MMF. Furthermore, this study clarifies the variations of glucose content in MMF during processing, which can not only indicate the processing degree but also provide a scientific basis for revealing the fermentation mechanism and improving the quality control of MMF.

[Quality evaluation of Psoraleae Fructus based on QAMS method and TLC identification].

The present study established a determination method of Psoraleae Fructus by quantitative analysis of multi-components by the single marker(QAMS) and further improved the thin-layer chromatography(TLC) method. The QAMS method was established by UPLC with psoralen as the internal marker, and the content of psoralenoside, isopsoralenoside, psoralen, and isopsoralen was simultaneously determined. As revealed by the comparison with results of the external standard method, the QAMS method was accurate and feasible. According to the current quality standards of Psoraleae Fructus, the TLC method was further optimized and improved, and bakuchiol was added for identification based on the original TLC method with psoralen and isopsoralen as indicators. This study provides a reference for improving the quality control method of Psoraleae Fructus.

Effects of Light Intensity and Spectral Composition on the Transcriptome Profiles of Leaves in Shade Grown Tea Plants (Camellia sinensis L.) and Regulatory Network of Flavonoid Biosynthesis.

Black net shade treatment attenuates flavonoid biosynthesis in tea plants, while the effect of light quality is still unclear. We investigated the flavonoid and transcriptome profiles of tea leaves under different light conditions, using black nets with different shade percentages, blue, yellow and red nets to alter the light intensity and light spectral composition in the fields. Flavonol glycosides are more sensitive to light intensity than catechins, with a reduction percentage of total flavonol glycosides up to 79.6% compared with 38.7% of total catechins under shade treatment. A total of 29,292 unigenes were identified, and the KEGG result indicated that flavonoid biosynthesis was regulated by both light intensity and light spectral composition while phytohormone signal transduction was modulated under blue net shade treatment. PAL, CHS, and F3H were transcriptionally downregulated with light intensity. Co-expression analysis showed the expressions of key transcription factors MYB12, MYB86, C1, MYB4, KTN80.4, and light signal perception and signaling genes (UVR8, HY5) had correlations with the contents of certain flavonoids (p < 0.05). The level of abscisic acid in tea leaves was elevated under shade treatment, with a negative correlation with TFG content (p < 0.05). This work provides a potential route of changing light intensity and spectral composition in the field to alter the compositions of flavor substances in tea leaves and regulate plant growth, which is instructive to the production of summer/autumn tea and matcha.

[Recent advances in treatment of viral pneumonia using Chinese patent medicine].

Viral pneumonia is caused by a spreading of lung infection caused by respiratory viruses. Some virus infections were found to be highly aggressive, leading to lung inflammation and severe damage in respiratory system with high fatality rate. Currently, there is no effective therapeutic drugs in the clinic. The common clinical symptoms of viral pneumonias include fever, rhinitis, runny nose, nonproductive cough, fatigue, myalgias and headaches after the immune system being tricked by driving cytokines and overactivated immune response induced by cytokine storms. Patients with severe symptoms could get persistent high fever, dysfunctional breathing, consciousness disorders and even respiratory failure, post-inflammatory pulmonary fibrosis, multi-organ damages, shock and so on. Most clinical treatments are used to inhibit virus replication, relieve symptoms, inhibit excessive inflammatory response, regulate immune balance and protect organs. Both applied and basic research demonstrate that Chinese patent medicine has certain anti-viral effects, effectively inhibiting viral pneumonia transiting from mild to severe, rapid relieving of patient symptoms because of their multi-component and multi-target integrated roles. This review has summarized the reports on the treatment of viral pneumonia. Based on the pathogenic characteristics of viral pneumonia, this paper summarizes the diverse roles of the marketed Chinese patent medicine, such as their effects in inhibiting the progress of viral replication and overactivated inflammatory response, regulating immune balance, attenuating pulmonary fibrosis and so forth. Our paper summarizes the advantages of Chinese patient medicine in the treatment of viral pneumonia, based on which improvements of clinical therapy are expected to be made soon.

Isopsoralen induces different subchronic toxicities and metabolomic outcomes between male and female Wistar rats.

Isopsoralen is a major active and quality-control component of Fructus Psoraleae, but lacks a full safety evaluation. We evaluated the oral toxicity of isopsoralen in Wistar rats treated for 3 months at doses of 0, 3.5, 7.0, and 14 mg/kg. Additionally, the plasma metabolomics of isopsoralen in male and female rats treated for 3 months at doses of 0 and 14 mg/kg were investigated by gas chromatography-mass spectrometry. Many abnormalities were observed in the isopsoralen-treated rats, including suppression of body weight gain, and changes in serum biochemical parameters and visceral coefficients. Histopathological changes in liver, pancreatic, and reproductive system tissues were also observed in the isopsoralen-treated rats. The metabolomic analyses showed alterations in many metabolites (19 in female rats; 28 in male rats) after isopsoralen administration. The significant changes in these metabolites revealed metabolomic alterations in the isopsoralen-treated rats, especially in amino acid metabolism regardless of sex, including phenylalanine, tyrosine, and tryptophan biosynthesis and glycine, serine, and threonine metabolism. Furthermore, fatty acid metabolism comprised the main affected pathways in female rats, while lipid metabolism and energy metabolism were the main affected pathways in male rats.

[Establishment pharmacological research platform for "Concurrent treatment of the brain and heart" and its application on dissecting mechanism of Danhong injection].

The Concurrent treatment of the brain and heart (CTBH) theory is proposed based on traditional Chinese medical theory and clinical practice. In this study, a framework for the pharmacological research platform was established to investigate the principles of concurrent treatment of the brain and heart. The platform for CTBH includes several key techniques for network modeling, discovery of active substances, dissecting mechanism of action and investigation of pharmacokinetic property of TCM. Taking network modeling of CTBH as an example, using database search, literature mining, network construction and module analysis, the that network modules closely associated with the pathological progress of cardiovascular and cerebrovascular diseases were identified, while further functional enrichment analysis of these modules indicated that the key biological processes included oxidative stress, metabolism and inflammation. GSK3B, NOTCH1, CDK4 were identified as key nodes in these network modules. The above-mentioned platform was applied to construct component-biomolecules network of Danhong injection for the identification of common targets and pathways. Among them, GSK3B had the highest correlation with the composition of Danhong injection in the network, and the biological function of whose cluster was related to cell oxidative stress. Based upon results of network analysis, validation experiments suggested that Danhong injection significantly improved the survival rate of oxidative injured myocardial cells and nerve cells, and the protective effect was related to the increase of phosphorylated GSK3ß protein expression. Moreover, extracts of Salviae Miltiorrhizae Radix et Rhizoma and Carthami Flos exerted the synergisticcytoprotective effect. The results indicated that the mechanism of treatment of cardiovascular and cerebrovascular diseases of Danhong injection could be studied through network modeling and other methods. In summary, the proposed pharmacological platform provided a feasible way for revealing the mechanism of CTBH by using modern scientific methods.

Danshen enhanced the estrogenic effects of Qing E formula in ovariectomized rats.

BACKGROUND: Menopause is characterized by a decrease in life quality due to the appearance of uncomfortable symptoms. Nowadays, Understanding menopause-associated pathophysiology and developing new strategies to improve the treatment of menopausal-associated symptoms is an important issue. Our study was to evaluate the synergistic effects of Danshen (salvia miltiorrhiza bunge) and the phytoestrogenic effects of 3 modified Qing E formulas, to explore a better formula for menopausal disorders. METHODS: 100 rats were randomized into 5 groups: Sham (Sham operation group), OVX (model group of ovariectomized rat), BDL (group with low concentration of Qing E Formula), BDH (group with high concentration of Qing E Formula) and BDD (group with high concentration of Qing E Formula Plus Danshen), receiving vehicle and extract of different modified Qing E formula respectively. The food intake, body weight, uterus weight, blood levels of triglycerides (TG), total cholesterol (TC) and cholesterol fractions were assessed. The mammary glands and uterus were morphologically analyzed. The bone density of tibias were measured by peripheral quantitative computed tomography (pQCT). Additionally, luciferase induction assays were performed in Hela cells with the mixtures derived from Qing E formula plus Danshen (BDD). RESULTS: Qing E formula plus Danshen significantly increased the uterus wet weight, enhanced the thickness of uterine wall, endometrial epithelium and glandular epithelium, improved trabecular bone and total density evidently, reduced the levels of low density lipoprotein cholesterol (LDL-C) and TG, possessed notable estrogen receptor beta (ERß) and estrogen receptor alpha (ERα) agonist activity. CONCLUSION: Qing E formula plus Danshen exerted more evident estrogen-like effects, thus it has a potential therapeutic use to treat menopausal disorders.

[Recent advances in anti-aging study of 2,3,5,4'-tetrahydroxystilbene-2-O-beta-D-glucopyranoside--a main component of Polygonum multiflorum].

Aging is an inevitable process of life caused by a combination of organs and tissues events which manifests as loss of structure and function. It is accompanied with organ hypofunction, decline in defense against stress and energy metabolism, and lots of age-related diseases. Therefore, it is critical to understand the mechanism of aging, as well as to discover new drugs to slow aging. Polygonum multiflorum has long been recognized in traditional Chinese medicine as anti-aging medicine. Interestingly, its extract has been shown to prolong lifespan in drosophila and elegans. Tetrahydroxystilbene glucoside (TSG), the main ingredient of P. multiflorum, shares a strong structural similarity with resveratrol, a well-known anti-aging natural product. This review summarized up-to-date literature reports and our laboratory findings on the mechanism of TSG anti-aging efficiency and function against age-related disease. It could provide reference for searching for anti-aging tradition Chinese medicine.

Phenolic Compounds from the Roots of Rhodiola crenulata and Their Antioxidant and Inducing IFN-γ Production Activities.

In the present study, two new phenolic compounds 1 and 11, a pair of lignan isomers 12 and 13 with their absolute configurations established for the first time, were isolated from the ethanol extract of the roots of Rhodiola crenulata, together with 13 known phenolic compounds, and their structures were elucidated via NMR, HRESIMS, UV, IR and CD analyses. All the isolated compounds were evaluated for their in vitro antioxidant activities using the 2,2-diphenyl-1-picryhydrazyl (DPPH) and 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging assays. Ten of them exhibited significant antioxidant activities compared to ascorbic acid. Furthermore, the inducibilities of the isolated compounds to IFN-γ production were also assessed. Compounds 1, 8, 9, 12, 13, 14 and 15 could moderately stimulate IFN-γ expression.

Characterization of in vivo antioxidant constituents and dual-standard quality assessment of Danhong injection.

Antioxidants and oxidative stress play a critical role in cardiovascular diseases. Danhong injection (DHI) is a well prescribed cardiovascular medication in China, but its detailed chemical basis and mechanisms of action remain unknown. To prove the antioxidant activity of DHI, its free radical scavenging capacity (RSC) was assessed by 2,2-diphenyl-1-picrylhydrazyl (DPPH) spectrophotometric assay. The 50% radical scavenging activity value was 1:129.2 mL/mL, against 0.95 mM DPPH. To further identify the antioxidant compounds, modified thin-layer chromatography combined with DPPH bioautography assay was used. Compared with vitamin C, 11 of 16 available compounds displayed strong antioxidant activity, which were also detected in rat serum after intravenous administration of DHI by ultra-performance liquid chromatography-tandem mass spectrometry, except for hydroxysafflor yellow A. Therefore, 10 antioxidants remaining in the blood as key markers, and six other compounds as general markers, were employed to perform the quality control of DHI by ultra-performance liquid chromatography-ultraviolet detection after systematic methodological validation. The analytical results indicate a high correlation (r = 0.9) between the total content of those antioxidants remaining in blood and RSC of DHI among 10 batches. Further, the antioxidant profiling and chemical marker quantification as dual-standard quality assessment was successfully applied to evaluate Danshen and safflower injections.

Intensive carbon combustion in sintering packed bed via steam spraying: An experimental study on carbon monoxide emission reduction.

Improving the combustion efficiency of solid fuels is important for reducing carbon monoxide emissions in the iron ore sintering process. In this paper, the surface steam spraying technology is introduced in the sintering process based on the auxiliary combustion effect of steam on coke, and its potential to reduce carbon monoxide emissions is demonstrated. Thermogravimetric analysis experiments of coke breeze in air and air-steam mixed atmosphere are carried out, and the results show that the introduction of steam can reduce the concentration of carbon monoxide in the exhaust gas from 183×10-6 to 78×10-6. At the same time, the mechanisms of carbon monoxide emission reduction by surface steam spraying technology are analyzed from the thermodynamic and kinetic perspectives. Then, a series of laboratory-scale sintering pot tests are carried out under no spraying operation, interval spraying operation, and continuous spraying operation. The results indicate that both interval and continuous spraying operations can reduce carbon monoxide emissions. The optimal mode of steam spraying under the present experimental conditions is continuously spraying for 13 min at a volume rate of 0.053 m3/min. Compared with no spraying, the average carbon monoxide concentration in the exhaust gas is reduced from 7565×10-6 to 6231×10-6, and total carbon monoxide emissions for per ton sinter are reduced from 13.46 m3/t to 9.51 m3/t.

Isolation and characterization of two new chroman-4-ones from the endophytic fungus Penicillium chrysogenum obtained from Eucommia ulmoides Oliver.

Two new chroman-4-ones penicichromanone A (1) and penicichromanone B (2), together with three known compounds conioxepinol C (3), emodin (4) and moniliphenone (5), were obtained from the endophytic fungus Penicillium chrysogenum, which was isolated from the bark of Eucommia ulmoides Oliver. The structures of 1 and 2 were elucidated by detailed analysis of HRESIMS, 1D/2D NMR and ECD spectra. All the compounds were evaluated for their anti-inflammatory activities using HEK293 cells, and compounds 1, 3, 4 and 5 exhibited significant inhibitory effects on TNF-α-stimulated NF-κB activation.

Ikaros is degraded by proteasome-dependent mechanism in the early phase of apoptosis induction.

Ikaros is an important transcription factor involved in the development and differentiation of hematopoietic cells. In this work, we found that chemotherapeutic drugs or ultraviolet radiation (UV) treatment could reduce the expression of full-length Ikaros (IK1) protein in less than 3h in leukemic NB4, Kasumi-1 and Jurkat cells, prior to the activation of caspase-3. Etoposide treatment could not alter the mRNA level of IK1 but it could shorten the half-life of IK1. Co-treatment with the proteasome inhibitor MG132 or epoxomicin but not calpain inhibitor calpeptin inhibited etoposide-induced Ikaros downregulation. Overexpression of IK1 could accelerate etoposide-induced apoptosis in NB4 cells, as evidenced by the increase of Annexin V positive cells and the more early activation of caspase 3. To our knowledge, this is the first report to show that upon chemotherapy drugs or UV treatment, IK1 could be degraded via the proteasome system in the early phase of apoptosis induction. These data might shed new insight on the role of IK1 in apoptosis and the post-translational regulation of IK1.

Aerobic denitrification performance and nitrate removal pathway analysis of a novel fungus Fusarium solani RADF-77.

Increasing nitrogenous contaminants have caused immense challenges to the environment and human health. As compared to physical and chemical methods, biological denitrification is considered to be an effective solution due to its environmental friendliness, high efficiency, and low cost. In the present work, a novel fungal strain identified as Fusarium solani (RADF-77) was isolated from cellulose material-supported denitrification reactor; this strain is capable of removing nitrogen under aerobic conditions. The average NO3--N removal rate for RADF-77 were 4.43 mg/(L·h) and 4.50 mg/(L·d), when using glucose and tea residue as carbon source, respectively. The nitrogen balance revealed that 53.66% of N vanished via gaseous products. Transcriptional results revealed that respiratory and assimilative nitrate reductases may work together for nitrate removal. Our results indicate that RADF-77 could be used as a potential means of enhancing nitrate-removal performance, as well as recycling tea residue, which is the main byproduct of the manufacture of tea extracts.

Protecting Intestinal Microenvironment Alleviates Acute Graft-Versus-Host Disease.

Acute gut graft-versus-host disease (aGVHD) is a leading threat to the survival of allogeneic hematopoietic stem cell transplantation (allo-HSCT) recipients. Abnormal gut microbiota is correlated with poor prognosis in allo-HSCT recipients. A disrupted intestinal microenvironment exacerbates dysbiosis in GVHD patients. We hypothesized that maintaining the integrity of the intestinal barrier may protect gut microbiota and attenuate aGVHD. This hypothesis was tested in a murine aGVHD model and an in vitro intestinal epithelial culture. Millipore cytokine array was utilized to determine the expression of proinflammatory cytokines in the serum. The 16S rRNA sequencing was used to determine the abundance and diversity of gut microbiota. Combining Xuebijing injection (XBJ) with a reduced dose of cyclosporine A (CsA) is superior to CsA alone in improving the survival of aGVHD mice and delayed aGVHD progression. This regimen also reduced interleukin 6 (IL-6) and IL-12 levels in the peripheral blood. 16S rRNA analysis revealed the combination treatment protected gut microbiota in aGVHD mice by reversing the dysbiosis at the phylum, genus, and species level. It inhibited enterococcal expansion, a hallmark of GVHD progression. It inhibited enterococcal expansion, a hallmark of GVHD progression. Furthermore, Escherichia coli expansion was inhibited by this regimen. Pathology analysis revealed that the combination treatment improved the integrity of the intestinal tissue of aGVHD mice. It also reduced the intestinal permeability in aGVHD mice. Besides, XBJ ameliorated doxorubicin-induced intestinal epithelial death in CCK-8 assay. Overall, combining XBJ with CsA protected the intestinal microenvironment to prevent aGVHD. Our findings suggested that protecting the intestinal microenvironment could be a novel strategy to manage aGVHD. Combining XBJ with CsA may reduce the side effects of current aGVHD prevention regimens and improve the quality of life of allo-HSCT recipients.

Ultra-low noise and high bandwidth bipolar current driver for precise magnetic field control.

Current sources with extremely low noise are significant for many branches of scientific research, such as experiments of ultra-cold atoms, superconducting quantum computing, and precision measurements. Here we construct and characterize an analog-controlled bipolar current source with high bandwidth and ultra-low noise. A precise and stable resistor is connected in series with the output for current sensing. After being amplified with an instrumentation amplifier, the current sensing signal is compared with an ultra-low noise reference, and proportional-integral (PI) calculations are performed via a zero-drift low-noise operational amplifier. The result of the PI calculation is sent to the output power operational amplifier for closed-loop control of the output current. In this way, a current of up to 16 A can be sourced to or sunk from a load with a compliance voltage of greater than ±12 V. The broadband current noise of our bipolar current source is about 0.5 µA/Hz and 1/f corner frequency is less than 1 Hz. Applications of this current source in a cold atom interferometer, as well as active compensation of a stray magnetic field, are presented. A method for measuring high-frequency current noise in a 10 A DC current with a sensitivity down to a level of 10 µA is also described.

Chemical constituents from the fruits of Psoralea corylifolia and their protective effects on ionising radiation injury.

Two new flavonoids, corylifol F (1) and corylifol G (2), together with 19 known compounds, were isolated from the fruits of Psoralea corylifolia L.. The structures of these compounds were determined by interpretation of spectroscopic data and comparison with literature properties. The radioprotective effects of the isolated compounds against ionising radiation damage were also evaluated in vitro. The results showed that corylifol A exhibited radioprotective effects in both HBL-100 and MCF-7 cells, while psoralen, isopsoralen, corylifol C and bakuchiol showed obvious selective action to protect HBL-100 cells against damage caused by ionising radiation.

[HPLC determination of four active components in dengzhanxixin injection].

OBJECTIVE: To establish a method for determination of chlorogenic acid, caffeic acid, caffeoylquinic acid and ScuteIlarin in Dengzhan Xixin injection. METHOD: The HPLC method was carried out on Agilent Zorbax SB-C18 column (150 mm x 4. 6 mm, 5 pim) evaluated with acetonitrile-0.1% formic acid as mobile phase, gradient elution; the flow rate was 1.0 mL x min(-1); the temperatue of column was at 35 degrees C; the detection wavelength was at 335 nm for UV detection. RESULT: The calibration curves were linear in the range of 0.025 to 0.800 microg (r = 0.9990) for chlorogenic acid, 0.027 to 0.850 microg (r = 0.9999) for caffeic acid, 0.062 to 1.978 microg (r = 0.9997) for caffeoylquinic acid and 0.118 to 3.770 microg (r = 0.9999) for scutellarin,respectilely. The average recoveries were 97.19% with RSD 1.16%; 100.45% with RSD 1.16%; 97.32% with RSD 1.43% and 103.81% with RSD 0.70%, respectively. CONCLUSION: The assay demonstrated that the method was simple, it had adequate accuracy and selectivity to quantify the four active components in Dengzhan Xixin injection.