Aberrant cerebral blood flow and functional connectivity in patients with vestibular migraine: a resting-state ASL and fMRI study.

BACKGROUND: Prior neuroimaging studies on vestibular migraine (VM) have extensively certified the functional and structural alterations in multiple brain regions and networks. However, few studies have assessed the cerebral blood flow (CBF) in VM patients using arterial spin labeling (ASL). The present study aimed to investigate CBF and functional connectivity (FC) alterations in VM patients during interictal periods. METHODS: We evaluated 52 VM patients and 46 healthy controls (HC) who received resting-state pseudo-continuous ASL and functional magnetic resonance imaging (fMRI) scanning. Comparisons of voxel-based CBF and seed-based FC were performed between the two groups. Brain regions showed significant group differences in CBF analyses were chosen as seeds in FC analyses. Additionally, the associations between abnormal imaging results and clinical features were explored. RESULTS: Compared with HC, VM patients showed higher normalized CBF in the right precentral gyrus (PreCG), left postcentral gyrus (PostCG), left superior frontal gyrus and bilateral insular (p < 0.05, FDR corrected). Furthermore, VM patients exhibited increased FC between the right PreCG and areas of the left PostCG, left cuneus and right lingual gyrus (p < 0.05, FDR corrected). In addition, we observed decreased FC between the left insular and regions of the left thalamus and right anterior cingulate cortex, as well as increased FC between the left insular and right fusiform gyrus in VM patients (p < 0.05, FDR corrected). Moreover, these variations in brain perfusion and FC were significantly correlated with multiple clinical features including frequency of migraine symptoms, frequency of vestibular symptoms and disease duration of VM (all p < 0.05). CONCLUSIONS: Patients with VM during interictal period showed hyperperfusion and abnormal resting-state FC in brain regions potentially contributed to disrupted multi-sensory and autonomic processing, as well as impaired ocular motor control, pain modulation and emotional regulation. Our study provided novel insights into the complex neuropathology of VM from a CBF perspective.

Characteristic volatile components analysis of Camelina sativa (L.) Crantz by GC-MS and their antibacterial and antioxidant activities.

Camelina sativa (L.) Crantz is an oilseed plant common in Europe and Asia. This study used the gas chromatography-mass spectrometry (GC-MS) to examine the differences in the aroma on the basis of extraction method such as water distillation extraction (CSPW), Solid-phase microextraction (CSPM) and subcritical extraction (CSPS). Antibacterial test was evaluated by the microdilution method against Salmonella typhimurium, Streptococcus pneumoniae, Escherichia coli, Strepococcus pyogenens, Staphylococcus aureus, and antioxidant activity was determined through DPPH free radical, hydroxyl free radical, and superoxide anion radical scavenging capacity activity. The result revealed that three extraction methods were distinct from each other based on their volatile compounds. Sixty-one volatiles of diverse chemical nature were identified and quantified. The volatile components contain thioether, aldehydes, alcohols, ketones, acids, esters, alkene, alkanes, amide, and furan compounds. The volatile components of Camelina sativa (L.) Crantz have good antibacterial and antioxidant activities. Furthermore, this work provides reference methods for detecting novel volatile organic compounds in plants and products.

Metabolomic analysis reveals the changing trend and differential markers of volatile and nonvolatile components of Artemisiae argyi with different aging years.

INTRODUCTION: Artemisia argyi Folium (AAF) is a traditional medicinal herb and edible plant. Analyzing the differential metabolites that affect the efficacy of AAF with different aging years is necessary. OBJECTIVE: The aim of the study was to investigate the changing trend and differential markers of volatile and nonvolatile metabolites of AAF from different aging years, which are necessary for application in clinical medicine. METHODOLOGY: Metabolites were analyzed using a widely targeted metabolomic approach based on ultrahigh-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) and gas chromatography tandem mass spectrometry (GC-MS). RESULTS: A total of 153 volatile metabolites and 159 nonvolatile metabolites were identified. Principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA) could clearly distinguish AAF aged for 1 year (AF-1), 3 years (AF-3), and 5 years (AF-5). Seven flavonoids and nine terpenoids were identified as biomarkers for tracking the aging years. CONCLUSIONS: The metabolomic method provided an effective strategy for tracking and identifying biomarkers of AAF from different aging years. This study laid the foundation for analysis of the biological activity of Artemisia argyi with different aging years.

Quality Evaluation of Peony Petals Based on the Chromatographic Fingerprints and Simultaneous Determination of Sixteen Bioactive Constituents Using UPLC-DAD-MS/MS.

In this study, a validated quality evaluation method with peony flower fingerprint chromatogram combined with simultaneous determination of sixteen bioactive constituents was established using UPLC-DAD-MS/MS. The results demonstrated that the method was stable, reliable, and accurate. The UPLC chemical fingerprints of 12 different varieties of peonies were established and comprehensively evaluated by similarity evaluation (SE), hierarchical cluster analysis (HCA), principal component analysis (PCA), and quantification analysis. The results of SE indicated that similar chemical components were present in these samples regardless of variety, but there were significant differences in the content of chemical components and material basis characteristics. The results of HCA and PCA showed that 12 varieties of samples were divided into two groups. Four flavonoids (11, 12, 13, and 16), five monoterpenes and their glycosides (3, 4, 6, 14, and 15), three tannins (7, 9, and 10), three phenolic acids (1, 2, and 5), and one aromatic acid (8) were identified from sixteen common peaks by standards and liquid chromatography-mass spectrometry (LC-MS). The simultaneous quantification of six types of components was conducted with the 12 samples, it was found that the sum contents of analytes varied obviously for peony flower samples from different varieties. The content of flavonoids, tannins, and monoterpenes (≥19.34 mg/g) was the highest, accounting for more than 78.45% of the total compounds. The results showed that the flavonoids, tannins, and monoterpenes were considered to be the key indexes in the classification and quality assessment of peony flower. The UPLC-DAD-MS/MS method coupled with multiple compounds determination and fingerprint analysis can be effectively applied as a feature distinguishing method to evaluate the compounds in peony flower raw material for product quality assurance in the food, pharmaceutical, and cosmetic industries. Moreover, this study provides ideas for future research and the improvement of products by these industries.

Astrocyte-specific knockout of YKL-40/Chi3l1 reduces Aß burden and restores memory functions in 5xFAD mice.

Glial cell-mediated neuroinflammation and neuronal attrition are highly correlated with cognitive impairment in Alzheimer's disease. YKL-40 is a secreted astrocytic glycoprotein that serves as a diagnostic biomarker of Alzheimer's disease. High levels of YKL-40 are associated with either advanced Alzheimer's disease or the normal aging process. However, the functional role of YKL-40 in Alzheimer's disease development has not been firmly established. In a 5xFAD mouse model of Alzheimer's disease, we observed increased YKL-40 expression in the cerebrospinal fluid of 7-month-old mice and was correlated with activated astrocytes. In primary astrocytes, Aß1-42 upregulated YKL-40 in a dose-dependent manner and was correlated with PI3-K signaling pathway activation. Furthermore, primary neurons treated with YKL-40 and/or Aß1-42 resulted in significant synaptic degeneration, reduced dendritic complexity, and impaired electrical parameters. More importantly, astrocyte-specific knockout of YKL-40 over a period of 7 days in symptomatic 5xFAD mice could effectively reduce amyloid plaque deposition in multiple brain regions. This was also associated with attenuated glial activation, reduced neuronal attrition, and restored memory function. These biological phenotypes could be explained by enhanced uptake of Aß1-42 peptides, increased rate of Aß1-42 degradation and acidification of lysosomal compartment in YKL-40 knockout astrocytes. Our results provide new insights into the role of YKL-40 in Alzheimer's disease pathogenesis and demonstrate the potential of targeting this soluble biomarker to alleviate cognitive defects in symptomatic Alzheimer's disease patients.

Metagenomics insights into gentamicin degradation and the dynamic of antibiotic resistance genes during co-composting of the gentamicin myelial residues with addition of various organic wastes.

Increasing antibiotic mycelial residues (AMRs) and related antibiotic resistance genes (ARGs) pose a significant threat to ecosystems and public health. Composting is a crucial method for recycling AMRs. However, the variation in ARGs and gentamicin degradation in the composting process of gentamicin mycelial residues (GMRs) has received little attention on an actual industrial scale. This research investigated the metabolic pathways and functional genes on the gentamicin and ARGs removal during the co-composting of GMRs with addition of various organic wastes (rice chaff, mushroom residue, etc.) under various C/N ratios (15:1, 25:1, 35:1). The results showed that the removal efficiencies of gentamicin and the total ARGs were 98.23 % and 53.20 %, respectively, with the C/N ratio of 25:1. Moreover, metagenomics and LS-MS/MS analysis demonstrated that the acetylation was the primary pathway for gentamicin biodegradation and the corresponding degrading genes were the categories of aac(3) and aac(6'). However, the relative abundance of aminoglycoside resistance genes (AMGs) were increased after 60 days composting. The partial least squares path modeling analysis demonstrated that the AMG abundance was directly influenced by the predominant mobile gene elements intI1 (p < 0.05) which was closely related to the bacterial community composition. Therefore, the ecological environmental risks should be assessed in the future application of GMRs composting products.

Near-infrared laser-irradiated upconversion nanoparticles with dexamethasone precise released for alleviating lung ischemia-reperfusion injury.

Introduction: Dexamethasone (DEX), as an important enduring-effect glucocorticoid (GC), holds great promise in the field of lung ischemia-reperfusion injury (LIRI) comprehensive therapy owing to its immunomodulatory properties, such as inducing apoptosis and cell cycle distribution. However, its potent anti-inflammatory application is still restricted because of multiple internal physiologic barriers. Methods: Herein, we developed upconversion nanoparticles (UCNPs) coated with photosensitizer/capping agent/fluorescent probe-modified mesoporous silica (UCNPs@mSiO2[DEX]-Py/ß-CD/FITC, USDPFs) for precise DEX release synergistic LIRI comprehensive therapy. The UCNPs were designed by covering an inert YOF:Yb shell on the YOF:Yb, Tm core to achieve high-intensity blue and red upconversion emission upon Near-Infrared (NIR) laser irradiation. Results: Under suitable compatibility conditions, the molecular structure of photosensitizer can be damaged along with capping agent shedding, which endowed USDPFs with an outstanding capability to carry out DEX release controlling and fluorescent indicator targeting. Furthermore, the hybrid encapsulating of DEX significantly increased utilization of nano-drugs, improving the water solubility and bioavailability, which was conducive to developing the anti-inflammatory performance of USDPFs in the complex clinical environment. Discussion: The response-controlled release of DEX in the intrapulmonary microenvironment can reduce normal cell damage, which can effectively avoid the side effects of nano-drugs in anti-inflammatory application. Meanwhile, the multi-wavelength of UCNPs endowed nano-drugs with the fluorescence emission imaging capacity in an intrapulmonary microenvironment, providing precise guidance for LIRI.

Oligosaccharides isolated from Rehmannia glutinosa protect LPS-induced intestinal inflammation and barrier injury in mice.

Objectives: We investigated the protective effect of Rehmannia glutinosa oligosaccharides (RGO) on lipopolysaccharide (LPS)-induced intestinal inflammation and barrier injury among mice. Methods: RGO is prepared from fresh rehmannia glutinosa by water extraction, active carbon decolorization, ion exchange resin impurity removal, macroporous adsorption resin purification, and decompression drying. LPS could establish the model for intestinal inflammation and barrier injury in mice. Three different doses of RGO were administered for three consecutive weeks. Then the weight, feces, and health status of the mice were recorded. After sacrificing the mice, their colon length and immune organ index were determined. The morphological changes of the ileum and colon were observed using Hematoxylin-eosin (H&E) staining, followed by measuring the villus length and recess depth. RT-qPCR was utilized to detect the relative mRNA expression of intestinal zonula occludens-1 (ZO-1) and occludin. The expression of inflammatory factors and oxidation markers within ileum and colon tissues and the digestive enzyme activities in the ileum contents were detected using ELISA. The content of short-chain fatty acids (SCFAs) in the colon was determined with GC. The gut microbial composition and diversity changes were determined with 16S-rRNA high-throughput sequencing. The association between intestinal microorganisms and SCFAs, occludins, digestive enzymes, inflammatory factor contents, and antioxidant indexes was also analyzed. Results: RGO significantly increased the weight, pancreatic index, thymus index, and colon length of mice compared with the model group. Moreover, it also improved the intestinal tissue structure and increased the expression of intestinal barrier-related junction proteins ZO-1 and Occludin. The contents of IL-6, IL-17, IL-1ß, and TNF-α in the intestinal tissues of mice were significantly reduced. Additionally, the activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and catalase (CAT) were elevated. In contrast, the malondialdehyde (MDA) content decreased. Trypsin and pancreatic lipase activities in the ileum enhanced, and the SCFA contents such as acetic acid, propionic acid, and butyric acid in the colon increased. The study on intestinal flora revealed that RGO could enhance the abundance of intestinal flora and improve the flora structure. After RGO intervention, the relative abundance of Firmicutes, Lactobacillus, and Akkermania bacteria in the intestinal tract of mice increased compared with the model group, while that of Actinomycetes decreased. The intestinal microbiota structure changed to the case, with probiotics playing a dominant role. The correlation analysis indicated that Lactobacillus and Ackermann bacteria in the intestinal tract of mice were positively associated with SCFAs, Occludin, ZO-1, pancreatic amylase, SOD, and CAT activities. Moreover, they were negatively correlated with inflammatory factors IL-6, IL-17, IL-1ß, and TNF-α. Conclusions: RGO can decrease LPS-induced intestinal inflammation and intestinal barrier injury in mice and protect their intestinal function. RGO can ameliorate intestinal inflammation and maintain the intestinal barrier by regulating intestinal flora.

Asymmetric Synthesis of Sterically Hindered 1-Substituted Tetrahydro-ß-carbolines Enabled by Imine Reductase: Enzyme Discovery, Protein Engineering, and Reaction Development.

We report the discovery of a new imine reductase (IRED), named AtIRED, by genome mining. Site-saturation mutagenesis on AtIRED generated two single mutants M118'L and P120'G and the double mutant M118'L/P120'G with improved specific activity toward sterically hindered 1-substituted dihydro-ß-carbolines. The synthetic potential of these engineered IREDs was showcased by the preparative-scale synthesis of nine chiral 1-substituted tetrahydro-ß-carbolines (THßCs), including (S)-1-t-butyl-THßC and (S)-1-t-pentyl-THßC, in 30-87% isolated yields with excellent optical purities (98-99% ee).

Quantitative fingerprint and antioxidative properties of Artemisia argyi leaves combined with chemometrics.

Quantitative fingerprint and differences of Artemisia argyi from different varieties, picking time, aging year, and origins were analyzed combing with chemometrics. The antioxidant activity was determined and antioxidant markers of Artemisia argyi were screened. Variety WA3 was significantly different from that of the other varieties. Fingerprint peak response and antioxidant activity of A. argyi picked in December were lower than samples collected in May and August. Fresh A. argyi leaves were significantly superior to withered leaves and stems. Artemisia argyi aging 1-5 years presented a classification trend. Antioxidant activity of A. argyi produced in Nanyang was generally superior to others origins. Peak 9, isochlorogenic acid A, and 6-methoxyluteolin contributed greatly for classification of A. argyi from different variety, picking time, aging year, and origin. Isochlorogenic acid A, isochlorogenic acid C, 6-methoxyluteolin, and chlorogenic acid were selected as antioxidant marker of A. argyi. The method based on quantitative fingerprint, antioxidant activity evaluation, and chemometrics was reliable to analyze the differences of A. argyi samples from different sources.

Qualitative and quantitative analysis of polyphenols in camelina seed and theirs antioxidant activities.

Camelina [Camelina sativa (L.) Crantz] seed has long been consumed as a source of food in Canada. But limited information is available concerning the systematical evaluation of the composition, content, and antioxidant activity of Camelina seed polyphenol extract (CSPE). Therefore, the aim of this study was to identify, quantify and evaluate the antioxidant activity of CSPE. The result showed that eight compositions were identified and determined by the UPLC-DAD-ESI-MS2 analysis. CSPE has potent free radical scavenging capacity. CSPE treatment significantly increased the activities of the antioxidant enzymes (superoxide dismutase and catalase) and glutathione content in a dose-dependent manner in RAW264.7 cells with oxidative injury and also reduced malondialdehyde content (P < 0.01). It may be concluded that CSPE has a strong antioxidant activity as depicted by the in vitro experiments and thus possesses the potential to be developed as food antioxidants or as an ingredient in functional foods.

Identification of Novel Artemisinin Hybrids Induce Apoptosis and Ferroptosis in MCF-7 Cells.

A series of novel 1,3,4-oxadiazole-artemisinin hybrids have been designed and synthesized. An MTT assay revealed that most of tested hybrids showed more enhanced anti-proliferative activities than artemisinin, among which A8 had the superior potency with IC50 values ranging from 4.07 µM to 9.71 µM against five tested cancer cell lines. Cell colony formation assays showed that A8 could inhibit significantly more cell proliferation than artemisinin and 5-fluorouracil. Further mechanism studies reveal that A8 induces apoptosis and ferroptosis in MCF-7 cells in a dose-dependent manner, and CYPs inhibition assays reveal that A8 has a moderate inhibitory effect on CYP1A2 and CYP3A4 in the human body at 10 µM. The present work indicates that hybrid A8 may merit further investigation as a potential therapeutic agent.

A Three-Dimensional Inversion Method for Small-Scale Magnetic Objects Based on Normalized Magnetic Source Strength.

The exploration of some dangerous or important small-scale magnetic objects requires accurate three-dimensional inversion results. In this paper, a three-dimensional inversion method for small-scale magnetic objects is proposed. Normalized magnetic source strength, which is weakly sensitive to the magnetization direction, is used for inversion, which avoids the influence of the remanence of magnetic objects. The planted inversion method is improved to make the inversion results more similar to the real results; normalized magnetic source strength is used to estimate the center position of the magnetic objects, which provides a priori information for the inversion; the weighting function is added in the inversion process to improve the inversion accuracy. The simulation and experimental results show that the method is not affected by the remanence, and effectively reduces the interference of non-target field sources. The obtained inversion results have higher accuracy.

A Small Target Localization Method Based on the Magnetic Gradient Tensor.

Currently, many small target localization methods based on a magnetic gradient tensor have problems, such as complex solution processes, poor stability, and multiple solutions. This paper proposes an optimization method based on the Euler deconvolution localization method to solve these problems. In a simulation, the Euler deconvolution method, an improved method of the Euler deconvolution method and our proposed method are analyzed under noise conditions. These three methods are evaluated in the field with complex magnetic interference in an experiment. The simulations show that the accuracy of the proposed method is higher than that of the improved Euler deconvolution method and is slightly lower for noisy conditions. The experimental results show that the proposed method is more precise and accurate than the Euler deconvolution and enhanced methods.

The mitotic regulator RCC2 promotes glucose metabolism through BACH1-dependent transcriptional upregulation of hexokinase II in glioma.

Weighted gene co-expression network analysis (WGCNA) identified a cell-cycle module that is associated with poor prognosis and aggressiveness of glioma. One of the core members, Regulator of chromatin condensation 2 (RCC2) is a component of the chromosome passenger complex. Accumulating evidence suggests that RCC2 plays a vital role in the mitotic process and that abnormal RCC2 expression is involved in cancer development. Gene silencing experiments show that RCC2 is required for glioma cell proliferation and migration. RNA-Sequencing analysis reveals a dual role of RCC2 in both the cell cycle and metabolism. Specifically, RCC2 regulates G2/M progression via CDC2 phosphorylation at Tyrosine 15. Metabolomic analysis identifies a role for RCC2 in promoting the glycolysis and pentose phosphate pathway. RCC2 exerts effects on metabolism by stabilizing the transcription factor BACH1 at its C-terminus leading to the transcriptional upregulation of hexokinase 2 (HK2). These findings elucidate a novel PTEN/RCC2/BACH1/HK2 signaling axis that drives glioma progression through the dual regulation of mitotic cell cycle and glycolytic events.

Analysis of Diagnostic Value of Electrogastrography for Parkinson's Disease and its Predictive Value for the Disease Progression.

The study aimed to evaluate the diagnostic and prognostic value of indexes detected by electrogastrography in Parkinson's disease patients. One hundred twenty early Parkinson's disease patients and 120 healthy controls were recruited, and underwent electrogastrography to detect dominant frequency (DF), instability coefficient of DF (ICDF), low frequency range (LFR), high frequency range (HFR), and normal frequency range (NFR). The receiver operating characteristic (ROC) curve was drawn for the diagnostic value analysis. The motor function was scored with the Unified Parkinson's Disease Rating Scale (UPDRS). Sniffin' Sticks test was used for the olfactory evaluation, and the TDI score consisting of odor threshold (T), odor discrimination (D) and odor identification (I) tests was calculated. The preprandial ICDF of Parkinson's disease patients was significantly higher than that of the control group, and decreased slowly during the late postprandial phase. The levels of LFR%, HFR% and NFR% in Parkinson's disease patients were higher than the control group during both the preprandial and late postprandial phase, and the changes of each index before and after meals were not obvious. Preprandial ICDF value and TDI score had the ability to distinguish Parkinson's disease patients with the AUC of 0.874 and 0.859 respectively. The ICDF detected by electrogastrography has high clinical value in the early diagnosis of Parkinson's disease, and the combination of ICDF and TDI can improve the diagnostic sensitivity and specificity of a single indicator. High ICDF levels during the preprandial phase are related to the poor prognosis of Parkinson's disease patients after treatment.

Application of Ketoreductase in Asymmetric Synthesis of Pharmaceuticals and Bioactive Molecules: An Update (2018-2020).

With the rapid development of genomic DNA sequencing, recombinant DNA expression, and protein engineering, biocatalysis has been increasingly and widely adopted in the synthesis of pharmaceuticals, bioactive molecules, fine chemicals, and agrochemicals. In this review, we have summarized the most recent advances achieved (2018-2020) in the research area of ketoreductase (KRED)-catalyzed asymmetric synthesis of chiral secondary alcohol intermediates to pharmaceuticals and bioactive molecules. In the first part, synthesis of chiral alcohols with one stereocenter through the bioreduction of four different ketone classes, namely acyclic aliphatic ketones, benzyl or phenylethyl ketones, cyclic aliphatic ketones, and aryl ketones, is discussed. In the second part, KRED-catalyzed dynamic reductive kinetic resolution and reductive desymmetrization are presented for the synthesis of chiral alcohols with two contiguous stereocenters.

New technology to improve the thermal stability of botulinum toxin type D by biomimetic mineralization.

The advanced biomimetic mineralization technology was applied to protect the Botulinum neurotoxin type D, and the processing of the mineralization granule of botulinum toxin type D was successfully screened. The loss of activity of the toxin protein at different temperatures and the destructive strength of the gastrointestinal tract against the toxin were determined biologically. The lethal toxicity of the mineralized toxin to wild rodents was determined by median lethal dose. Protective tests at different temperatures showed that the preservation period of botulinum toxin type D mineralized sample 2 was significantly higher than that of the control group at three different temperatures, and its toxicity loss was significantly reduced. The damage intensity of the mineralized toxin to the gastrointestinal contents of plateau zokor and plateau pika was significantly reduced. The minimum lethal doses of the mineralized toxin particles to plateau zokor, plateau pika, and mice were 5200, 8,600,000, and 25,000 MLD/kg. These results showed that biomimetic mineralization could greatly improve the thermal stability of botulinum toxin type D and reduce the damaging effect of the gastrointestinal contents of target animals to botulinum toxin type D. The mineralized toxin could be used to control the population density of urban rodents. This research provides new insights into the protection of toxin protein substances.

Butylphthalide soft capsules combined with modified tonic exercise therapy on neurological function and ability of daily living of patients with stroke hemiplegia.

OBJECTIVE: To explore the effect of butylphthalide soft capsules combined with modified tonic exercise therapy on neurologic function and the abilities of daily living in patients with stroke hemiplegia. METHODS: In this retrospective trial, a total of 90 patients with stroke hemiplegia admitted to our hospital from January 2019 to January 2020 were enrolled and divided into a control group and an experimental group according to different treatment methods. The two groups were both treated with butylphthalide soft capsules, and the experimental group was additionally treated by modified tonic exercise therapy. The clinical efficacy, endothelial injury indicators, molecular indicators of oxidative stress, and adverse reactions of the two groups were compared. Generic Quality of Life Inventory-74 (GQOLI-74) was used to assess the quality of life of patients after treatment. The Fugl-Meyer Upper Extremity Assessment (FMA) was used to evaluate their limb function before and after treatment, the National Institute of Health Stroke Scale (NIHSS) was used to evaluate their brain nerve function before and after the treatment, and the activities of daily living (ADL) were employed to assess their activities of daily living before and after treatment. RESULTS: After treatment, the experimental group outperformed the control group in terms of total clinical efficacy (P<0.05). The experimental group had significantly lower endothelial injury indicators and higher molecular indicators of oxidative stress than the control group (all P<0.001). The incidence of adverse reactions in the experimental group was lower than that in the control group (P<0.05). Higher GQOLI-74, FMA, and ADL scores and a significantly lower NIHSS score were obtained in the experimental group than the control group after treatment (P<0.001). CONCLUSION: For patients with stroke hemiplegia, butylphthalide soft capsule combined with modified tonic exercise therapy effectively improves their neurologic function, abilities of daily living, and quality of life.

Asymmetric Synthesis of a Key Dextromethorphan Intermediate and Its Analogues Enabled by a New Cyclohexylamine Oxidase: Enzyme Discovery, Reaction Development, and Mechanistic Insight.

(S)-1-(4-Methoxybenzyl)-1,2,3,4,5,6,7,8-octahydroisoquinoline [(S)-1-(4-methoxybenzyl)-OHIQ, (S)-1a] is a key synthetic intermediate in the industrial production of dextromethorphan, one of the most widely used over-the-counter antitussives. We report here that a new cyclohexylamine oxidase discovered by genome mining, named CHAOCCH12-C2, was able to completely deracemize 100 mM 1a under Turner's deracemization conditions to afford (S)-1a in 80% isolated yield and 99% ee at a semipreparative scale (0.4 mmol). When this biocatalytic reaction was scaled up to a gram scale (5.8 mmol), without reaction optimization (S)-1a was still isolated in 67% yield and 96% ee. The relatively higher kcat determined for CHAOCCH12-C2 was rationalized as one major factor rendering this enzyme capable of oxidizing 1a effectively at elevated substrate concentrations. Protein sequence alignment, analysis of our co-crystal structure of CHAOCCH12-C2 complexed with the product 1-(4-methoxybenzyl)-3,4,5,6,7,8-hexahydroisoquinoline [1-(4-methoxybenzyl)-HHIQ, 2a], and the structure-guided mutagenesis study together indicated L295 is one of the critical residues for this efficient enzymatic oxidation process and supported the presence of two cavities as well as a catalytically important "aromatic cage" formed by F342, Y433, and FAD. The synthetic applicability of CHAOCCH12-C2 was further underscored by the stereoselective synthesis of various enantioenriched 1-benzyl-OHIQ derivatives of potential pharmaceutical importance at a semipreparative scale.