Structure-Based Discovery of the SARS-CoV-2 Main Protease Noncovalent Inhibitors from Traditional Chinese Medicine.

Traditional Chinese medicine (TCM) has been extensively employed for the treatment of coronavirus disease 2019 (COVID-19) caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). However, there is demand for discovering more SARS-CoV-2 Mpro inhibitors with diverse scaffolds to optimize anti-SARS-CoV-2 lead compounds. In this study, comprehensive in silico and in vitro assays were utilized to determine the potential inhibitors from TCM compounds against SARS-CoV-2 Mpro, which is an important therapeutic target for SARS-CoV-2. The ensemble docking analysis of 18263 TCM compounds against 15 SARS-CoV-2 Mpro conformations identified 19 TCM compounds as promising candidates. Further in vitro testing validated three compounds as inhibitors of SARS-CoV-2 Mpro and showed IC50 values of 4.64 ± 0.11, 7.56 ± 0.78, and 11.16 ± 0.26 µM, with EC50 values of 12.25 ± 1.68, 15.58 ± 0.77, and 29.32 ± 1.25 µM, respectively. Molecular dynamics (MD) simulations indicated that the three complexes remained stable over the last 100 ns of production run. An analysis of the binding mode revealed that the active compounds occupy different subsites (S1, S2, S3, and S4) of the active site of SARS-CoV-2 Mpro via specific poses through noncovalent interactions with key amino acids (e.g., HIS 41, ASN 142, GLY 143, MET 165, GLU 166, or GLN 189). Overall, this study provides evidence indicating that the three natural products obtained from TCM could be further used for anti-COVID-19 research, justifying the investigation of Chinese herbal medicinal ingredients as bioactive constituents for therapeutic targets.

Antiplatelet effect of ginkgo diterpene lactone meglumine injection in acute ischemic stroke: A randomized, double-blind, placebo-controlled clinical trial.

This study was designed to evaluate antiplatelet effect and therapeutic effect of ginkgo diterpene lactone meglumine injection (GDLI) in acute ischemic stroke (AIS) patients. In this randomized, double-blind, placebo-controlled trial, we randomly assigned 70 inpatients within 48 hr after the onset of AIS to combination therapy with GDLI and aspirin (GDLI at a dose of 25 mg/d for 14 days plus aspirin at a dose of 100 mg/d for 90 days) or to placebo plus aspirin in a ratio of 1:1. Platelet function, the National Institute of Health Stroke Scale (NIHSS), and the modified Rankin Scale (mRS) were evaluated. A good outcome was defined as NIHSS scores decrease ≥5 or mRS scores decrease ≥2. Results showed that arachidonic acid induced maximum platelet aggregation rate (AA-MAR) and mean platelet volume (MPV) of the GDLI-aspirin group were much lower than that of the aspirin group (p = 0.013 and p = 0.034, respectively) after the 14-day therapy. The combination of GDLI and aspirin was superior to aspirin alone, and had significant impact on the good outcome at day 90 (ORadj 7.21 [95%CI, 1.03-50.68], p = 0.047). In summary, GDLI has antiplatelet effect and can improve the prognosis of AIS patients.

Identification of Prognostic LncRNAs Subtypes Predicts Prognosis and Immune Microenvironment for Glioma.

Glioma is the most commonly occurring primary neuroepithelial neoplasm. Long noncoding RNAs (lncRNAs) are emerging as pivotal modulators of gene expression in the immune system and play critical roles in the growth, progression, and immune response of carcinomas. In this study, we performed univariate Cox regression analysis on survival data from TCGA and identified 20 prognostic lncRNAs. Moreover, we revealed that these prognosis-related lncRNAs (PRLnc) were dysregulated in glioma. Furthermore, we constructed a signature based on the expression levels of these prognosis-related lncRNAs based on 13 prognostic lncRNAs, including AGAP2-AS1, CYTOR, MIR155HG, LINC00634, HOTAIRM1, SNHG18, LINC01841, LINC01842, LINC01426, MIR9-3HG, TMEM220-AS1, LINC00641, LINC01270, and LINC01503. The Kaplan-Meier curves show that high-risk patients had a shorter survival time. Finally, the glioma samples were classified into 2 subgroups based on the median expression of prognosis-related lncRNAs in each sample. In summary, these findings suggest that PRLnc is associated with tumor-infiltrating immune cells in glioma and that subtype 2 patients may respond more positively to immunotherapy.

Folic acid Ameliorates the Declining Quality of Sodium Fluoride-Exposed Mouse Oocytes through the Sirt1/Sod2 Pathway.

Excessive sodium fluoride (NaF) intake interferes with reproductive function in humans and animals; however, strategies to prevent these effects are still underexplored. Here, we showed that in vivo and in vitro supplementation of folic acid (FA) efficaciously improved the quality of NaF-exposed oocytes. FA supplementation not only increased ovulation of oocytes from NaF-treated mice but also enhanced oocyte meiotic competency and fertilization ability by restoring the spindle/chromosome structure. Moreover, FA supplementation could exert a beneficial effect on NaF- exposed oocytes by restoring mitochondrial function, eliminating reactive oxygen species accumulation to suppress apoptosis. We also found that FA supplementation restored the defective phenotypes in oocytes through a Sirt1/Sod2-dependent mechanism. Inhibition of Sirt1 with EX527 abolished the FA-mediated improvement in NaF-exposed oocyte quality. Collectively, our data indicated that FA supplementation is a feasible approach to protect oocytes from NaF-related deterioration.

Noni (Morinda citrifolia L.) fruit phenolic extract supplementation ameliorates NAFLD by modulating insulin resistance, oxidative stress, inflammation, liver metabolism and gut microbiota.

The liver-protective activity of phenolics has been consistently reported, but the underlying protective mechanism of phenolic extract from noni fruit (NFE) against high-fat-diet (HFD)-induced nonalcoholic fatty liver disease (NAFLD) remains unclear. Mice were fed with HFD or combination of HFD and NFE for 10 weeks, and then the gut microbiota and liver metabolites were compared. In this study, NFE supplementation alleviated HFD-induced liver injury and metabolic comorbidities, as evidenced by reduced liver function markers, decreased lipid profile levels, and improved obesity and insulin resistance. NFE supplementation restored the composition of gut microbiota with a remarkable elevation in the relative abundance of Parabacteroides, Lactobacillus, Roseburia, Akkermansia and a significant reduction in Helicobacter, norank_f_Desulfovibrionaceae, Desulfovibrio, Mucispirillum at the genus level. Liver metabolomics demonstrated that NFE supplementation favorably regulated the metabolic pathways involved in oxidative stress and inflammation, including purine metabolism, glutathione metabolism, primary bile acid biosynthesis, glycerophospholipid metabolism, pentose phosphate pathway, ascorbate and aldarate metabolism, galactose metabolism etc. Furthermore, NFE supplementation inhibited the HFD-induced activation of the liver endotoxin - TLR4 - NF-κB pathway, and alleviated liver inflammation. In conclusion, the findings of this study provide new evidences supporting that NFE can be used as a therapeutic approach for HFD-induced NAFLD via modulating the gut microbiota composition, liver metabolite profile and suppressing inflammatory reaction.

Cardioprotective effect of anisodamine against ischemia/reperfusion injury through the mitochondrial ATP-sensitive potassium channel.

Previous clinical studies have shown that anisodamine could improve no-reflow phenomenon and prevent reperfusion arrhythmias, but whether this protective effect is related to the antagonism of the M-type cholinergic receptor or other potential mechanisms is uncertain. The aim of the present study was to investigate the role of the mitochondrial ATP-sensitive potassium channel (mitoK ATP ) in cardioprotective effect of anisodamine against ischemia/reperfusion injury. Anisodamine and 5- hydroxydecanoic acid were used to explore the relationship between anisodamine and mitoK ATP . Using a Langendorff isolated heart ischemia/reperfusion injury model, hemodynamic parameters and reperfusion ventricular arrhythmia were evaluated; in addition, changes in myocardial infarct size, cTnI from coronary effluent and myocardial ultrastructure, as well as ATP, MDA and SOD in myocardial tissues, were detected. In the hypoxia/reoxygenation injury model of neonatal rat cardiomyocyte, cTnI release in the culture medium and levels of ATP, MDA and SOD in cardiomyocytes and mitochondrial membrane potential, were analyzed. Overall, anisodamine could significantly improve the hemodynamic indexes of isolated rat heart injured by ischemia/reperfusion, reduce the occurrence of ventricular reperfusion arrhythmia and myocardial infarction area, and improve the ultrastructural damage of myocardium and mitochondria. The in vitro results demonstrated that anisodamine could improve mitochondrial energy metabolism, reduce oxidative stress and stabilize mitochondrial membrane potential. The cardioprotective effects were significantly inhibited by 5-hydroxydecanoic acid. In conclusion, this study suggests that the opening of mitoK ATP could play an important role in the protective effect of anisodamine against myocardial ischemia/reperfusion injury.

Novel Bithiophene Dimers from Echinops latifolius as Potential Antifungal and Nematicidal Agents.

Three novel dimeric bithiophenes, echinbithiophenedimers A-C (1-3), along with two known thiophenes, 4 and 5, were obtained from Echinops latifolius, and their structures were identified through extensive spectroscopic analysis and electronic circular dichroism calculations. Compounds 1-3 possessed new carbon skeletons; they are dimeric bithiophenes with 1 and 2 featuring an unprecedented 1,3-dioxolane ring system and 3 featuring an unusual 1,4-dioxane ring. These compounds are the first examples of bithiophene dimers furnished by different cyclic diethers. Dimeric bithiophenes 1-3 had good antifungal activities against five phytopathogenic fungi, and compound 3 showed excellent activity against Alternaria alternate and Pyricularia oryzae, with a minimal inhibitory concentration value of 8 µg/mL, which was close to or higher than that of carbendazim. Moreover, its effect on the mycelial morphology was observed by scanning electron microscopy. Compounds 1-3, which were demonstrated to be nonphototoxic thiophenes, exhibited better nematicidal activity than the commercial nematicide ethoprophos against Meloidogyne incognita. This study revealed that dimeric bithiophenes containing 1,3-dioxolane or 1,4-dioxane rings could be used as novel antifungal and nematicidal agents for controlling plant fungal and nematode pathogens.

Sesquiterpenoids from Artemisia vestita and Their Antifeedant and Antifungal Activities.

Four new sesquiterpenoids, named artemivestinolide D-G (1-4) and three known sesquiterpenoids (5-7), were isolated from Artemisia vestita. The structures of these new compounds were determined based on extensive spectroscopic data analyses. Furthermore, the electronic circular dichroism data determined the absolute configurations of the new compounds. The antifeedant and antifungal activities of the isolates were evaluated against third-instar larvae of Plutella xylostella and three plant pathogenic fungi. Compounds 1-7 showed moderate antifeedant activities and compounds 1-4 and 6-7 exhibited antifungal activities.

Thiophenes from Echinops grijsii as a Preliminary Approach To Control Disease Complex of Root-Knot Nematodes and Soil-Borne Fungi: Isolation, Activities, and Structure-Nonphototoxic Activity Relationship Analysis.

Naturally occurring thiophenes possess excellent nematicidal and fungicidal activities. However, thiophenes often have limited application in soil due to their light-dependent toxicity given the living and reproductive condition of soil-borne pathogens. In this study, six new (1-6) and six known thiophenes (7-12) were isolated from Echinops grijsii. Compounds 1-2, 4-5, 8-9, 11 , and 12 showed stronger nematicidal activity against Meloidogyne incognita than commercial nematicide abamectin. 4-10 were demonstrated as nonphototoxic thiophenes. Among these, 4 and 8 were the most potent thiophenes (LC50 values 2.57 and 0.91 µg/mL in light, 1.80 and 0.86 µg/mL in dark, respectively) against M. incognita. SAR revealed that thiophene skeleton was essential for nematicidal activity, while disubstituted groups were helpful for nonphototoxicity. Although an increased number of acetylenes improved activity, it decreased nonphototoxicity. Acyl groups could suppress the effects of light on activity, with the level of inhibitory effects depending on its number and chain length, while chlorine played important roles in promoting activity. Additionally, compounds 1-2, 4-5, 7, 8, and 10 displayed antifungal activity against six soil-borne fungi in various degrees. The discovery of nonphototoxic thiophenes and elucidation of SAR provide important information for the exploitation and utilization of thiophenes in the integrative management regarding disease complexes caused by the combination of root-knot nematode and soil-borne fungi.

Characterization of the chemical constituents in Hongjingtian injection by liquid chromatography quadrupole time-of-flight mass spectrometry.

Hongjingtian injection is made from Rhodiola wallichiana and used in the treatment of stable angina pectoris associated with coronary heart disease. In this study, the chemical constituents in Hongjingtian injection were comprehensively studied using liquid chromatography quadrupole time-of-flight mass spectrometry. A total of 49 compounds were identified or assumed, including 10 organic acids, nine phenylethanoids, 10 phenylpropanoids, two flavonoid glycosides, seven monoterpene glycosides, seven octylglycosides and four other types of compounds. The structures of seven compounds were confirmed by comparing their retention times, MS and UV spectra with the corresponding authentic standards. Amongst the 49 compounds, 35 were firstly found in R. wallichiana, while they have been reported in other species of the genus Rhodiola, including Rhodiola crenulata, Rhodiola sacra, Rhodiola rosea and Rhodiola kirilowii. The possible fragmentation pathways in the mass spectrometry of the major types of compounds are proposed and summarized. Our study demonstrates a rapid method for characterizing the chemical constituents present in the Hongjingtian injection, which could also be applied to the identification of chemical constituents in other TCM formulae containing R. wallichiana.

[Effect of Acupuncture Combined with Rehabilitation on Immune and Neural Functions in Elderly Patients with Stroke].

OBJECTIVE: To investigate the therapeutic effect of acupuncture combined with rehabilitation and neuro-immune functional activities in elderly patients with stroke. METHODS: A total of 196 elderly stroke patients were randomly divided into control (rehabilitation exercise) and observation(acupuncture＋ rehabilitation exercise)groups (n＝98 in each). Patients of the control group were treated by general healing treatment including good limb placement, timely conversion of body position, joint-motor exercise, sitting balance exercise, sitting-standing-walking, walking up and down stairs, daily life ability exercise, etc. and those of the observation group were treated by using the same methods mentioned in the control group and manual acupuncture stimulation of Sishencong (EX-HN 1), Xuanli (GB 6), the midpoint of EX-HN 1 and GB 6, Baihui (GV 20), Qubin (GB 7), and the midpoint between GV 20 and GB 7 on the affected side (once a day for three 10-day courses). Serum interleukin-2 (IL-2), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), and high-sensitive C-reactive protein (hs-CRP) contents were assayed using ELISA, and serum CD 3＋， CD 4＋ and CD 8＋ contents assayed using flow cytometry. The disability severity was assessed by using National Institutes of Health Stroke Scale (NIHSS, for dysneuria), modified Rankin Scale，(mRS, stroke severity), and Barthel Index (BI, performance in activities of daily living), separately. The therapeutic effect was determined according to NIHSS score, clinical symptoms and daily activity ability. RESULTS: On day 30 after the treatment, serum IL-2, IL-6, TNF-α and hs-CRP contents were significantly decreased in the observation group in comparison with its own pre-treatment and day 10 after the treatment, and on day 10 and 30 after the treatment, the above were lower than the control group (P<0.05). On day 30 after the treatment, serum CD 3＋， CD 4＋ and CD 4＋/CD 8＋ levels were significantly increased in both control and observation groups in comparison with their own pre-treatment and 10 days' treatment (P<0.05), while CD 8＋ levels obviously decreased in both groups relevant to their own pre-treatment and 10 days' treatment (P<0.05), and the increased CD 3＋， CD 4＋ and CD 4＋/CD 8＋ levels were significant higher in the observation group than in the control group (P<0.05), and the CD 8＋ level was obviously lower in the control group than in the observation group (P<0.05). On day 30 after the treatment, the NIHSS and mRS scores were significantly decreased in both groups (P<0.05), and significantly lower in the observation group than in the control group (P<0.05); the BI scores were evidently increased in both groups relevant to their own pre-treatment (P<0.05), and the BI level was considerably higher in the observation group than in the control group (P<0.05). The total effective rate was 72.4% (71/98) and 93.9% (92/98) respectively in the control and observation groups, being obviously higher in the latter group than in the former (P<0.05). CONCLUSION: Acupuncture combined with rehabilitation therapy is effective in promoting the recovery of neurological function and in regulating T lymphocyte subsets and the expression of inflammatory factors in elderly patients with stroke.

Oleanane-Type Saponins from the Roots of Ligulariopsis shichuana and Their α-Glucosidase Inhibitory Activities.

Five new oleanane-type saponins, named ligushicosides A-E, and three known oleanane-type saponins were isolated from the roots of Ligulariopsis shichuana. Their structures were established by a combination of spectroscopic techniques, including 1D and 2D NMR and high resolution electrospray ionization mass spectroscopy (HR-ESI-MS). Furthermore, all isolates were evaluated for their yeast α-glucosidase inhibitory effects and exhibited potent inhibition against α-glucosidase, while compounds 1 and 2 showed excellent inhibitory activities. The 3-O-glycoside moiety in oleanane-type saponin is important for the α-glucosidase inhibitory effects.

Hongjingtian Injection Attenuates Myocardial Oxidative Damage via Promoting Autophagy and Inhibiting Apoptosis.

Natural products with antioxidative activities are widely applied to prevent and treat various oxidative stress related diseases, including ischemic heart disease. However, the cellular and molecular mechanisms of those therapies are still needed to be illustrated. In this study, we characterized the cardioprotective effects of Hongjingtian Injection (HJT), an extensively used botanical drug for treating coronary heart disease. The H/R-induced profound elevation of oxidative stress was suppressed by HJT. HJT also attenuates oxidative injury by promoting cell viability, intracellular ATP contents, and mitochondrial oxygen consumption. Validation experiments indicated that HJT inhibited H/R-induced apoptosis and regulated the expression of apoptosis-associated proteins Bcl-2 and cleaved caspase3. Interestingly, HJT significantly regulated the expression of autophagy-related proteins LC3, Beclin, and mTOR as well as ERK and AKT. We provide evidence that the mechanism involves activation of AKT/Beclin-1, AKT, and ERK/mTOR pathway in cardiomyocyte autophagy. Histological and physiological evaluation revealed that HJT significantly decreased the infarct area of the heart, improved cardiac function, and increased the expression of LC3B in a rat model of coronary occlusion. From the obtained data, we proposed that HJT diminished myocardial oxidative damage through regulating the balance of autophagy and apoptosis and reducing oxidative stress.

Schizophrenia affects speech-induced functional connectivity of the superior temporal gyrus under cocktail-party listening conditions.

The superior temporal gyrus (STG) is involved in speech recognition against informational masking under cocktail-party-listening conditions. Compared to healthy listeners, people with schizophrenia perform worse in speech recognition under informational speech-on-speech masking conditions. It is not clear whether the schizophrenia-related vulnerability to informational masking is associated with certain changes in FC of the STG with some critical brain regions. Using sparse-sampling fMRI design, this study investigated the differences between people with schizophrenia and healthy controls in FC of the STG for target-speech listening against informational speech-on-speech masking, when a listening condition with either perceived spatial separation (PSS, with a spatial release of informational masking) or perceived spatial co-location (PSC, without the spatial release) between target speech and masking speech was introduced. The results showed that in healthy participants, but not participants with schizophrenia, the contrast of either the PSS or PSC condition against the masker-only condition induced an enhancement of functional connectivity (FC) of the STG with the left superior parietal lobule and the right precuneus. Compared to healthy participants, participants with schizophrenia showed declined FC of the STG with the bilateral precuneus, right SPL, and right supplementary motor area. Thus, FC of the STG with the parietal areas is normally involved in speech listening against informational masking under either the PSS or PSC conditions, and declined FC of the STG in people with schizophrenia with the parietal areas may be associated with the increased vulnerability to informational masking.

Two-stage integrated care versus antipsychotic medication alone on outcomes of schizophrenia: One-year randomized controlled trial and follow-up.

Integrated care can reduce rate of relapse and improve personal and social functions in patients with schizophrenia. We established and evaluated a new model of "intensive-consolidation" two-stage integrated care (IC) for inpatients with schizophrenia. Data were collected between 2012 and 2015. Chinese inpatients with schizophrenia (n=170) diagnosed according to DSM-IV were randomly assigned to antipsychotic medication-alone (n=84) or two-stage IC (n=86) and followed up for 12 months. The IC model included intensive treatments (antipsychotics plus the cognitive behavior therapy and rehabilitation treatment) during hospitalization and 3-time consolidation treatments with 3-month intervals at clinics. Outcome measures included the rate of relapse, psychiatric symptoms and social functioning. Compared with medication-alone group, the rate of relapse were significantly lower in IC group (p=0.012); the Mixed-Effects Model for Repeated-Measures analyses showed that the IC group significantly improved in positive symptoms over time; greater improvement in self-care and less aggressive behaviors were observed over time in IC group (all p<0.008). The findings support the feasibility and effectiveness of the new two-stage model of integrated care as an intervention for middle-acute-phase inpatients with schizophrenia. The model is particularly informative to countries where medical resources are mainly distributed in developed regions.

[Anti-tumor metastatic constituents from Rhodiola wallichiana].

To study the anti-tumor metastatic constituents in Rhodiola wallichiana (HK) S H Fu var Cholaensis (Praeg) S H Fu, chemical constituents were isolated and purified by repeated column chromatography (silica gel, Toyopearl HW-40C and preparative HPLC). Their structures were elucidated on the basis of spectral data analysis. The anti-tumor metastasis assay was applied to evaluate the activities of the isolated compounds. Ten compounds (1-10) were isolated and their structures were identified by comparison of their spectral data with literature as follows: syringic acid (1), salidroside (2), tyrosol (3), scaphopetalone (4), berchemol (5), 2,6-dimethoxyacetophenone (6), rhobupcyanoside A (7), miyaginin (8), chavicol-4-O-ß-D-apiofuranosyl-(1 --> 6)-O-ß-D-glucopyranoside (9), eugenyol-O-ß-D-apiofuranosyl-(1 --> 6)-O-ß-D-glucopyranoside (10). Compounds 4-6 and 8-10, were isolated from this genus for the first time, while compound 7 was isolated from this plant for the first time. Compounds 2, 6-8 showed positive anti-tumor metastatic activities, and compounds 2 and 8 showed significant anti-tumor metastatic activities.

[Eudesmane sesquiterpenes from twigs of Manglietia hookeri].

Chemical constituents from the acetone extract of twigs of Manglietia hookeri were isolated and purified by various column chromatographic methods over silica gel and sephadex LH-20, and preparative TLC. The structures of these compounds were identified on the basis of physicochemical properties and spectral analysis, including NMR and MS spectra. Six eudesmane sesquiterpenes were obtained and their structures were identified as trans-eudesmane-4, 11-diol(1), ß-eudesmol(2), (-) -10-epi-5ß-hydroxy-ß-eudesmol (3), epi-carrisone (4), 6-hydroxy-eudesm-4(14) -ene(5) and gynurenol(6). All the compounds were isolated from this plant for the first time. Furthermore, the 13C-NMR data of compound 3 were reported for the first time.

A new illicinolide from leaves of Illicium micranthum Dunn.

The ethyl acetate extract from leaves of Illicium micranthum Dunn exhibited radical scavenging ability on DPPH radical-scavenging assay. Further phytochemical investigation led to the isolation of a new sesquiterpenoid, illicinolide C (1), and eight known compounds (2-9). The structural elucidations of the compounds were based on spectroscopic methods and by comparison of their spectroscopic data with those reported in the references. Except compound 9, all the other compounds were isolated from I. micranthum Dunn for the first time, while compounds 1 and 2 were new to the Illiciaceae family.

6,8-Dihy-droxy-8a-methyl-3,5-dimethyl-idenedeca-hydro-naphtho-[2,3-b]furan-2(3H)-one.

The title compound, C(15)H(20)O(4), is a eudesmanolide isolated from the Chinese medicinal plant Carpesium tris-te Maxim. The mol-ecule contains three rings, viz. two fused six-membered rings in chair conformations and a five-membered ring in a flattened envelope conformation. In the crystal, two hy-droxy groups are involved in the formation of intra- and inter-molecular O-H⋯O hydrogen bonds. The H atoms in these groups are split, with site-occupation factors of 0.5. The inter-molecular hydrogen bonds link mol-ecules into chains propagating in [010].