IFITM3 restricts porcine deltacoronavirus infection by targeting its Spike protein.

The discovery of antiviral molecules is crucial for controlling porcine deltacoronavirus (PDCoV). Previous studies have provided evidence that the IFN-inducible transmembrane protein 3 (IFITM3), which is coded by an interferon-stimulated gene, prevents the infections of a number of enveloped viruses. Nevertheless, the involvement of IFITM3 in PDCoV infection remains unexplored. In this study, it was observed that the overexpression of IFITM3 successfully restrictes the infection of PDCoV in cell cultures. Conversely, the suppression of IFITM3 facilitates the infection of PDCoV in IPI-2I and IPEC-J2 cells. Further studies revealed that IFITM3 limits the attachment phase of viral infection by interacting with the S1 subunit of the PDCoV Spike (S) protein. In addition, IFITM3 is verified as a member of the CD225 family, the GxxxG conserved motif of this family is important for it to limit PDCoV infection. In summary, this study reveals the mechanism of IFITM3 as an antiviral molecule to inhibit PDCoV infection, and also provides theoretical supports for screening effective anti-PDCoV drugs.

Can digital finance reduce carbon emission intensity? A perspective based on factor allocation distortions: evidence from Chinese cities.

The world is facing the challenges of climate change and energy structure adjustments. The role of digital finance, a new branch of business that combines digital technology and traditional financial products, in reducing global carbon emissions needs to be studied. This paper uses panel data on 280 cities in China from 2011 to 2019 to empirically examine the efficacy of digital finance for governing carbon emission reductions and the mechanism by which it does so. The results show that (1) digital finance can facilitate carbon emission reductions and help reduce carbon emission intensity within regions; (2) digital finance helps promote the rational allocation of resources and alleviates factor distortions by encouraging firms to rationally use their own factor endowments so as to reduce carbon emission intensity, which holds robustly after considering the endogenous issues such as possibly omitting variables and collinearity; and (3) differences in geographical location, the vitality of regional innovation and entrepreneurship, regional willingness to protect the environment, and environmental protection levels lead to heterogeneity in the effect of digital finance on carbon emission intensity. Therefore, it is necessary to vigorously develop digital finance as a long-term tool for carbon governance.

Comparative Investigation of Raw and Processed Radix Polygoni Multiflori on the Treatment of Vascular Dementia by Liquid Chromatograph-Mass Spectrometry Based Metabolomic Approach.

Radix Polygoni Multiflori (PM) is a well-known nootropic used in traditional Chinese medicine (TCM). Considering the efficacy and application discrepancy between raw (RPM) and processed PM (PPM), the similarities and differences between them in the treatment of vascular dementia (VaD) is intriguing. In this study, a VaD rat model was constructed by 2-vessel occlusion (2-VO). During 28 days of treatment, plasma was collected on days 7, 14, 21, and 28 after the start of dosing and the metabolic profile was analyzed by HPLC-MS/MS-based metabolomics. The Morris Water Maze Test, hematoxylin-eosin and Nissl staining, and biochemical analysis were used to assess cognitive function, pathogenic alterations and oxidative stress, respectively. RPM and PPM effectivelyreducedthe 2VO-induced cognitive impairment and mitigated histological alterations in hippocampus tissue. The 2-VO model significantly elevated MDA level and decreased SOD activity and GSH level, indicating severe oxidative stress, which could also be attenuated by RPM and PPM treatment. RPM outperformed PPM in decreasing MDA levels while PPM outperformed RPM in increasing GSH levels. Differential metabolites were subjected to Metabolite Set Enrichment Analysis (MSEA) and genes corresponding to proteins having interactions with metabolites were further annotated with Gene Ontology (GO). Both RPM and PPM ameliorated VaD-relevant vitamin B6 metabolism, pentose phosphate pathways, and taurine and hypotaurine metabolism. In addition, the metabolism of cysteine and methionine was regulated only by RPM, and riboflavin metabolism was modulated only by PPM. The results suggested that raw and processed PM had comparable efficacy in the treatment of VaD but also with some mechanistic differenece.

Grossamide attenuates inflammation by balancing macrophage polarization through metabolic reprogramming of macrophages in mice.

Macrophages exhibited different phenotypes in response to environmental cues. To meet the needs of rapid response to stimuli, M1-activated macrophages preferred glycolysis to oxidative phosphorylation (OXPHOS) in mitochondria to quickly produce energy and obtain ample raw materials to support cell activation at the same time. Activated macrophages produced free radicals and cytokines to eradicate pathogens but also induced oxidative damage and enhanced inflammation. Grossamide (GSE), a lignanamide from Polygonum multiflorum Thunb., exhibited notable anti-inflammatory effects. In this study, the potential of GSE on macrophage polarization was explored. GSE significantly down-regulated the levels of M1 macrophage biomarkers (Cd32a, Cd80 and Cd86) while increased the levels of M2 indicators (Cd163, Mrc1 and Socs1), showing its potential to inhibit LPS-induced M1 polarization of macrophages. This ability has close a link to its effect on metabolic reprogramming of macrophage. GSE shunted nitric oxide (NO) production from arginine by up-regulation of arginase and down-regulation of inducible nitric oxide synthase, thus attenuated the inhibition of NO on OXPHOS. LPS created three breakpoints in the tricarboxylic acid cycle (TCA) cycle of macrophage as evidenced by down-regulated isocitrate dehydrogenase, accumulation of succinate and the inhibited SDH activity, significantly decreased level of oxoglutarate dehydrogenase expression and its substrate α-ketoglutarate. Thus GSE reduced oxidative stress and amended fragmented TCA cycle. As a result, GSE maintained redox (NAD+/NADH) and energy (ATP/ADP) state, reduced extracellular acidification rate and enhanced the oxygen consumption rate. In addition, GSE decreased the release of inflammatory cytokines by inhibiting the activation of the LPS/TLR4/NF-κB pathway. These findings highlighted the central role of immunometabolism of macrophages in its functional plasticity, which invited future study of mode of action of anti-inflammatory drugs from viewpoint of metabolic reprogramming.

Pharmacokinetics and metabolism of olerciamide A from Portulaca oleracea L. in rats by UHPLC-UV and UHPLC-ESI-Q-TOF/MS.

The aim of this study was to elucidate the pharmacokinetics of olerciamide A in rats after oral and intravenous administration of Portulaca oleracea L. extract by a simple and rapid ultra high-performance liquid chromatography method with bergapten as internal standard. The pharmacokinetic results indicated that olerciamide A was rapidly distributed with a time to peak concentration of 30 min after oral administration and presented a low oral absolute bioavailability of 4.57%. The metabolism of olerciamide A in rats was also investigated using ultra-high-performance liquid chromatography electrospray coupled with quadrupole-time of flight mass spectrometry to elucidate the reason for the low absolute bioavailability of olerciamide A and seven metabolites of oleraciamide A were found in rat plasma and urine.

Two new similar alkaloids from Portulaca oleracea L.

Two novel alkaloids named oleraciamide A (1) and oleraciamide B (2) were isolated from Portulaca oleracea L., and spectroscopic methods including 1D and 2D nuclear magnetic resonance and high-resolution electrospray ionisation quadrupole-time of flight mass spectrometer spectrometry techniques are employed to determine their structures. Oleraciamide A (1) was evaluated no cytotoxicity at concentrations up to 80 µM over 72 h against human adipose-derived stem cells (hADSCs) by CCK-8 method.

Correction to Three Novel Alkaloids from Portulaca oleracea L. and Their Anti-inflammatory Effects.

Oleracimine and oleracimine A were isolated from Portulaca oleracea L. and described in the J. Agric. Food Chem, but the alternative structures of the two compounds are proposed on the basis of NMR analyses.

Three Novel Alkaloids from Portulaca oleracea L. and Their Anti-inflammatory Effects.

Three novel carbon skeleton alkaloids, named oleracimine (1), oleracimine A (2), and oleracone A (3), with one novel azulene carbon skeleton compound, oleracone B (4), and one known compound, ß-carboline (5), were first isolated from Portulaca oleracea L. The structures were determined using spectroscopic methods, including one- and two-dimensional nuclear magnetic resonance and high-resolution electrospray ionization time-of-flight mass spectrometry techniques. In addition, oleracimine (1) was used to investigate the anti-inflammatory effects on lipopolysaccharide-stimulated macrophages. The results of enzyme-linked immunosorbent assay, western blot, and real-time polymerase chain reaction showed that oleracimine (1) remarkably inhibited nitric oxide production and could dose-dependently decrease the secretions of interleukin 6, tumor necrosis factor α, nitric oxide, and prostaglandin E2 in cell culture supernatants as well as the mRNA of cyclooxygenase-2 and inducible nitric oxide synthase.

Polarity, selectivity and performance of hydrophilic organic/salt-containing aqueous two-phase system on counter-current chromatography for polar compounds.

The essential attributes of a solvent system for separation polar compounds on CCC are polarity, selectively and performance. Here, hydrophilic organic/salt-containing aqueous two-phase system (HO/S TPS) was evaluated as an alternative solvent system for CCC separation of polar compounds. Polarity measurements based on Rohrschneider-Snyder parameter was developed as quantitative assessing the polarity of HO/S TPS and comparing with an organic/aqueous system. All investigated 1-butanol/ethanol/saturated ammonium sulfate solution/water (BEAsWat) and 1-butanol/ethanol/saturated dipotassium hydrogen phosphate solution/water (BEDhpWat) systems with polarity values of organic phase from 4.5 to 6.8, were more polar than chloroform/methanol/water (1/1/1). The considerable water content of BEAsWat and BEDhpWat (0/1/1/1/) was 45.4 and 42.6% (w%) of hydrophilic organic phase, and 66.4 and 51.2% (w%) of salt-containing aqueous phase, respectively, closed to conventional aqueous two-phase system. Therefore, the polarity of HO/S TPS is in the middle of organic/aqueous and aqueous two-phase system. The LogKC values of twenty four polar compounds as model mixture confirmed that the polarities of HO/S TPSs were matched to that of the polar compounds and shown to be a very selective technique capable of separating positional isomers. Moreover, BEAsWat and BEDhpWat systems can be easily retained in CCC column with suitable elution mode. The hydrodynamic behavior reversion of HO/S TPS on hydrodynamic CCC was observed and was tentatively explained based on the density difference. Finally, caffeoylquinic acid isomers and dihydroxybenzoic acid isomers were successfully separated with HO/S TPS on CCC, respectively. Those results demonstrate that HO/S TPS on CCC is a performant and stable way to separate polar compounds from natural products.

Cytotoxic biflavones from Stellera chamaejasme.

Bioassay-guided phytochemical studies on Stellera chamaejasme led to the isolation of two new biflavones, chamaejasmenin E (1) and chamaejasmin D (2), together with ten known compounds. The structures of new compounds were elucidated by extensive spectroscopic analyses and their absolute configurations on 2, 3, 2″ and 3″ were confirmed by TDDFT quantum chemical calculated ECD spectra combined with experimental ECD spectra. All isolated biflavones were evaluated for their cytotoxic activities against Bel-7402 and A549 tumor cell lines, and sikokianin D (3) was found to possess the most potential cytotoxic activities against both the two cell lines with IC50 values of 1.29 ± 0.21 and 0.75 ± 0.25 µM, respectively. Moreover, some structure-function relationships of these bioflavones for cytotoxic activities were explored and summarized.

Hydrophilic organic/salt-containing aqueous two-phase solvent system for counter-current chromatography: a novel technique for separation of polar compounds.

Hydrophilic organic/salt-containing aqueous two-phase system composing of ethanol, water and ammonium sulfate for separation polar compounds was investigated on multilayer coil associated with J-type HSCCC devices. Compared to the classical polar solvent system based on 1-butanol-water or PEG1000-ammonium sulfate-water, the water content of upper phase in ethanol-ammonium sulfate-water systems was from 53.7% to 32.8% (wt%), closed to PEG1000-ammonium sulfate-water aqueous two-phase systems and higher than 1-butanol-water (22.0%, wt%). Therefore, the polarity of ethanol-ammonium sulfate-water is in the middle of 1-butanol-water and PEG-ammonium sulfate-water system, which is quite good for separating polar compounds like phenols, nucleosides and amino acids with low partition coefficient in 1-octanol-water system. The retention of stationary phase in four elution mode on type-J counter-current chromatography devices with multilayer coil column changed from 26% to 71%. Hydrodynamic trend possess both intermediate and hydrophilic solvent system property, which closely related to the composition of solvent system. The applicability of this system was demonstrated by successful separation of adenosine, uridine guanosine and cytidine.

A novel preparative liquid chromatograph for repetitive enrichment and purification of low-abundance compounds.

A novel recycle preparative HPLC system was developed to repeatedly enrich and purify low-abundance compounds from complex samples. The recycle separation involves injecting a large volume of the sample through a trap column, releasing the absorbed sample for preparative HPLC separation, and then capturing the target component in the same trap column for the next separation. After several cycles, the final purified target compound absorbed in the trap column is eluted with a small volume of solvent. The capture and recycle procedure was realized by adjusting the strength of the mobile phase using an auxiliary pump and switching the two valves connecting the trap and preparative columns. Compared with standard HPLC or existing recycle chromatography, this system not only reduces the solvent side-effects and sample volume overloading during injection but also avoids peak-broadening and sensitivity loss during the recycle runs. Moreover, the final purified products can be easily concentrated. With the aid of this system, a 10% polyphenol in extracts from Saussurea involucrata cultured cells reached >95% purity with more than 95% recovery after three recycle purifications, and a 1% unstable epimer impurity in the raw material of the drug silybin reached >95% purity with 75% recovery after a six-cycle separation.

Activity-guided isolation of antioxidant compounds from Rhizophora apiculata.

Rhizophora apiculata (R. apiculata) contains an abundance of biologically active compounds due its special salt-tolerant living surroundings. In this study, the total phenolic content and antioxidant activities of various extract and fractions of stem of R. apiculata were investigated. Results indicated that butanol fraction possesses the highest total phenolic content (181.84 mg/g GAE/g dry extract) with strongest antioxidant abilities. Following in vitro antioxidant activity-guided phytochemical separation procedures, lyoniresinol-3α-O-ß-arabinopyranoside (1), lyoniresinol-3α-O-ß-rhamnoside (2), and afzelechin-3-O-L-rhamno-pyranoside (3) were separated from the butanol fraction. These compounds showed more noticeable antioxidant activity than a BHT standard in the DPPH, ABTS and hydroxyl radical scavenging assays. HPLC analysis results showed that among different plant parts, the highest content of 1-3 was located in the bark (0.068%, 0.066% and 0.011%, respectively). The results imply that the R. apiculata might be a potential source of natural antioxidants and 1-3 are antioxidant ingredients in R. apiculata.

A new diterpenoid from Rhizophora apiculata.

A new diterpenoid, 15(S)-isopimar-7-en-1-oxo-15,16-diol (1), was isolated from the stems of mangrove plant Rhizophora apiculata. The structure of the new compound was elucidated by MS, IR, 1D, and 2D NMR techniques, including HMQC, HMBC, and NOESY correlations. In addition, seven known constituents were isolated from this plant for the first time.

[Preparation of flavonol glycoside reference standard series from Epimedium brevicornum Maxim using pilot-scale preparative high performance liquid chromatography].

Icariin and epimedins A, B, C are a series of active flavonol glycoside in Epimedium brevicornum Maxim. A pilot-scale preparative high performance liquid chromatographic (HPLC) method was developed to purify the four flavonol glycosides as reference standards from the crude extract of Epimedium brevicornum Maxim. After the crude extract containing approximately 20% flavonols was enriched using macropore resin, the obtained target fractions were subjected to pilot-scale preparative HPLC purification. With the aid of a self-packed pilot-scale preparative column (220 mm x 77 mm, 10 microm), the 4 target compounds were separated well within 35 min in a single chromatographic run by the elution with acetonitrile-water (26:74 or 30:70, v/v). By repetitive injection of the enriched target fraction onto the preparative column, 33 g icariin as well as 4.6 g epimedin C, 3.7 g epimedin B, and 0.6 g epimedin A were obtained from 300 g crude extract. The purities of all products were greater than 98%. This pilot-scale preparative HPLC technique and the two step separation technology for 4 target compounds are quite useful for the production of the reference standard series with good purity like icariin, epimedins A, B, C standards due to its high performance, rapid separation and more amounts of products obtained.

[Preparation of gentiopicroside from the extract of Swertia mussotii Franch by preparative high performance liquid chromatography].

A new method for isolation of gentiopicroside (GPS) from Swertia mussotii Franch with reversed phase preparative high performance liquid chromatography (Pre-HPLC) is described. After 95% ethanol extract was obtained with solvent extraction and macroporous resin separation, a fraction containing GPS was acquired and subjected to Pre-HPLC for isolation of target component. The optimum operation parameters were selected as follows: a C18 column (200 mm x 50 mm, 5 microm), methanol-0.1% acetic water (30:70, v/v) as the mobile phase at flow rate of 75 mL/min, the detection wavelength at 254 nm, and the injection volume of 500 microL at the mass concentration of 225 g/L. The purity of the product was detected by HPLC method. The result showed that the purity of product was above 99%. The effective and rapid method has been successfully applied to the preparation of GPS from Swertia mussotii Franch.

[Optimization of the collection region in preparative high performance liquid chromatography with exponentially modified Gaussian (EMG) model].

Preparative high performance liquid chromatography (HPLC) are widely used recently. The optimization of its operating conditions is important for improving efficiency and saving operating cost. Now, most works such as linear scale up technology were focused on the optimization of mobile phase. The sample collection region is another important operating parameter in preparative HPLC. In this study, a software was written according to the exponentially modified Gaussian (EMG) model. With this software, the operators of preparative HPLC can easily choose a suitable sample collection region.

[Analysis of chlorogenic acids in Helianthus tuberosus Linn leaves using high performance liquid chromatography-mass spectrometry].

To identify chlorogenic acids in Helianthus tuberosus Linn leaves, a method of high performance liquid chromatography-ultraviolet-mass spectrometry (HPLC-UV-MS) was developed. HPLC analysis was performed on an Inertsil ODS-3 column (250 mm x 4.6 mm, 5 microm). The mobile phase consisted of 1% acetic acid (A) and methanol (B). A gradient program was adopted as follows: 0 - 10 min, 20% B to 35% B; 10 - 25 min, 35% B to 50% B; 25 - 35 min, 50% B to 80% B. The flow rate was set at 1.0 mL/min and the column temperature was 35 degrees C. HPLC chromatogram was extracted at 327 nm. The mass spectrometer used was a TSQ triple quadrupole MS equipped with an electrospray ionization (ESI) interface. Initially, the mass spectrometer was programmed to perform full scan ranging from m/z 250 - 1 200. For MS/MS, negative ion monitoring mode was used and the collision energy was set at 10 - 45 eV. By analyzing UV characteristics and MS fragmentation patterns, 7 chlorogenic acids were assigned to be three categories, i. e., three caffeoylquinic acids, one feruloylquinic acid and three dicaffeoylquinic acids. As the results of the method development efforts, an effective and fast method for the qualitative identification of the chlorogenic acids in Helianthus tuberosus Linn leaves was established.

[Preparation of asiaticoside and madecassoside from the extract of Centella asiatica (L.) Urb using preparative high performance liquid chromatography].

A high performance liquid chromatographic (HPLC) method was successfully used to separate asiaticoside and madecassoside from the extract of Centella asiatica (L.) Urb. The raw material containing 20% asiaticoside and 45% madecassoside was dissolved in methanol and used for the preparation of the pure compounds. The flow rate of the mobile phase, injection volume and detection wavelength were optimized. The optimum operation parameters were then selected as follows: a C18 column (50 mm x 200 mm, 5 microm) with methanol-water (60 : 40, v/v) as the mobile phase at a flow rate of 100 mL/min, the detection wavelength of 220 nm, and the injection volume of 1.5 mL at the concentration of 40 mg/mL. Asiaticoside and madecassoside the can be obtained in a 20 min runtime, and the purities of the products were detected by melting point, thin layer chromatography (TLC) and HPLC method. The results showed that the purities of products were above 98%. The effective and rapid method has been successfully applied and to the preparation of asiaticoside and madecassoside from Centella asiatica (L. ) Urb.

[Multicenter clinical trial on the efficacy of thrombolytic therapy with urokinase and/or anticoagulant with low molecular weight heparin in acute pulmonary embolism].

OBJECTIVE: To evaluate the efficacy of thrombolytic therapy with urokinase and/or anticoagulant with low molecular weight heparin in patients with acute pulmonary embolism. METHODS: 127 patients with documented acute pulmonary embolism were treated with different regimens: thrombolysis combined with anticoagulant therapy in 70 cases, thrombolytic therapy alone in 31 cases and pure anticoagulant therapy in 26 cases. RESULTS: Dyspnea was the most common symptom (91.3%), then cough (74.0%), chest pain (63.0%) and syncope (21.3%). As compared with pre-treatment in each group, there was significant improvement in respiratory rate, heart rate, partial pressure of oxygen in artery and partial pressure of carbon dioxide in artery. The effective rate of thrombolysis combined with anticoagulant therapy, thrombolytic therapy and anticoagulant therapy were 90.0%, 77.4% and 61.5%, respectively, the total effective rate of thrombolytic therapy was 86.1%. The best efficacy was demonstrated in patients within one week after onset of symptoms. The therapy might be effective for cases with duration longer than two weeks after the onset of symptoms. CONCLUSION: Thrombolytic therapy with urokinase combined with low molecular weight heparin is effective and safe for acute pulmonary embolism.