To investigate the effects of phosphorus fertilizer on the morphological traits, active ingredients and rhizosphere soil microbial community of Polygala tenuifolia. The phosphorus fertilizer was calculated in terms of P_2O_5. Five treatments were set up: 0(CK), 17(P1), 34(P2), 51(P3), and 68(P4) kg per Mu(1 Mu≈667 m~2). A randomized block design was adopted. Samples of P. tenuifolia and its rhizosphere soil were collected under different superphosphate fertilizer treatments. Illumina high-throughput sequencing was used to analyze the rhizosphere soil microbial community, 9 morphological traits were measured and the content of 11 active ingredients were determined. The results showed that the whole plant weight, shoot fresh weight, root weight, and root peel thickness were the highest under P1 treatment, increasing by 34.41%, 38.80%, 39.21%, and 3.17% respectively compared to CK. Under P2 treatment, the plant height, stem diameter, root thickness, and core thickness were significantly higher than CK. Phosphorus fertilizer had a significant impact on the content of tenuifolin, sibiricose A5, sibiricose A6, arillanin A, 3,6'-disinapoyl sucrose, and polygalaxanthone Ⅲ. Correlation analysis results showed that the relative abundance of Arthrobacter, Bacillus, norank＿f＿Vicinamibacteraceae, norank＿o＿Vicinamibacterales, MND1 and other bacteria, as well as the relative abundance of Neocosmospora, Paraphoma and other fungi were positively correlated with root diameter, wood core diameter, the whole plant weight, root weight, shoot fresh weight of P. tenuifolia. Bacillus, Neocosmospora, Subulicystidium were significantly positively correlated with oligosaccharides such as 3,6'-disinapoyl sucrose, sibiricose A5、sibiricose A6、glomeratose A、arillanin A and tenuifoliside C. Arthrobacter, Humicola, Aspergillus, Paraphoma were positively correlated with tenuifolin and norank＿f＿Vicinamibacteraceae, norank＿o＿Vicinamibacterales, Fusarium were positively correlated with polygalaxanthone Ⅲ. Evidently, appropriate phosphorus application is conducive to the growth and quality improvement of P. tenuifolia, and can increase the abundance of beneficial microorganisms in the soil.
Rhizosphere ; Phosphorus/pharmacology* ; Soil Microbiology ; Polygala/anatomy & histology* ; Fertilizers/analysis* ; Bacteria/metabolism* ; Soil/chemistry* ; Microbiota/drug effects* ; Plant Roots/metabolism*

ObjectiveTo observe the effects of Hedysarum polysaccharides（HPS）on the signaling pathways of B-cell lymphoma 2 （Bcl-2）， cysteinyl aspartate-specific protease 3 （Caspase-3）， and Bcl-2-associated X protein （Bax） in Schwann cells（SCs）cultured in high glucose，and explore the possible mechanism of HPS against diabetic peripheral neuropathy（DPN）. MethodFour SD suckling mice aged 5-7 days were randomly divided into a normal group，a high-glucose group，an HPS + high-glucose group，and an α-lipoic acid（α-LA）+ high-glucose group. SCs were extracted from the sciatic nerve and cultured in a 37 ℃，5% CO₂ incubator. After the cells reached 80% confluence，Cell Counting Kit-8（CCK-8）was used to screen the experimental concentrations suitable for high glucose，HPS， and α-LA interventions. Western blot and Real-time polymerase chain reaction （Real-time PCR）were used to detect the protein and mRNA expression of Bcl-2，Bax，and Caspase-3. The apoptosis rate of SCs was detected by flow cytometry using Annexin V-fluorescein isothiocyanate （FITC）/propidium iodide （PI）. ResultAs revealed by Western blot and real-time PCR，compared with the normal group，the high-glucose group showed reduced protein and mRNA expression of Bcl-2 and increased protein and mRNA expression of Bax and Caspase-3（P<0.01）. Compared with the high-glucose group，the HPS + high-glucose group and the α-LA + high-glucose group showed increased protein and mRNA expression of Bcl-2 and decreased protein and mRNA expression of Bax and Caspase-3（P<0.01）. As displayed by the results of flow cytometry using Annexin V/PI， compared with the normal group，the high-glucose group showed increased apoptosis rate；compared with the high-glucose group，the HPS + high-glucose group and the α-LA + high-glucose group showed reduced apoptosis rate（P<0.01）. ConclusionHPS can alleviate the apoptotic response of SCs，and its mechanism may be related to the inhibition of the activation of the Bcl-2/Caspase-3 signaling pathway.

Objective To evaluate the effects of early exercise intervention in patients who have undergone primary isolated valve surgery. Methods Forty patients scheduled for mitral, aortic, and/or tricuspid valve surgery were allocated to receive a supervised exercise intervention consisting of cycling for 3 min/d at the bedside after operation (intervention group, n=20, mean age [49.05±3.728] years) or to receive no exercise intervention (control group, n=20, mean age [47.95 ± 3.214] years). Oxygen saturation (SpO2) was measured by pulse oximetry continuously before and after the 6-minute walk test. Psycho-educational counseling was provided, and patients were assessed using standard patient questionnaires. Results The arterial SpO2 level increased significantly in the intervention group after exercise compared with the control group (P<0.05). Heart rate returned to baseline in the intervention group postoperatively and was significantly lower than that in the control group (P<0.05). Conclusions A small amount of supervised cycling exercise at the bedside is a safe activity that may improve peripheral arterial SpO2 and reduce heart rate to the baseline level following longer distance walk before discharge in patients who have undergone isolated valve surgery.

Coronavirus disease 2019(COVID-19),which is caused by SARS-CoV-2,varies with regard to symptoms and mortality rates among populations.Humoral immunity plays critical roles in SARS-CoV-2 infection and recovery from COVID-19.However,differences in immune responses and clinical features among COVID-19 patients remain largely unknown.Here,we report a database for COVID-19-specific IgG/IgM immune responses and clinical parameters(named COVID-ONE-hi).COVID-ONE-hi is based on the data that contain the IgG/IgM responses to 24 full-length/truncated proteins corresponding to 20 of 28 known SARS-CoV-2 proteins and 199 spike protein peptides against 2360 serum samples collected from 783 COVID-19 patients.In addition,96 clinical parameters for the 2360 serum samples and basic information for the 783 patients are integrated into the database.Furthermore,COVID-ONE-hi provides a dashboard for defining samples and a one-click analysis pipeline for a single group or paired groups.A set of samples of interest is easily defined by adjusting the scale bars of a variety of parameters.After the"START"button is clicked,one can readily obtain a comprehensive analysis report for further interpretation.COVID-ONE-hi is freely available at www.COVID-ONE.cn.

OBJECTIVES: To explore the possible mechanism of Yunaconitine poisoning by studying the changes of urine metabolic profile in rats chronically poisoned by Yunaconitine via non-targeted metabolomics. METHODS: A rat model of Yunaconitine poisoning was established, and a metabolomics method based on UPLC-QTOF-MS technology was used to obtain the urine metabolic profile. Principal component analysis (PCA), orthogonal projections to latent structures-discriminant analysis (OPLS-DA), variable importance in projection (VIP) value greater than 1, fold change (FC) value greater than 3 or less than 0.33 and P value less than 0.05 were used to screen potential biomarkers related to the toxicity of Yunaconitine. The metabolic pathway analysis was performed through the MetaboAnalyst website and pathological changes of related tissues were observed. RESULTS: Sixteen potential biomarkers including L-isoleucine were screened, which mainly involved six metabolic pathways including the biosynthesis and degradation of valine, leucine and isoleucine, pentose and glucuronate interconversions, and propanoate metabolism, alanine, aspartate and glutamate metabolism, tyrosine metabolism. Pathological studies showed that rat toxic change in nervous system, liver and cardiac caused by Yunaconitine. CONCLUSIONS Yunaconitine may cause neurotoxicity, hepatotoxicity and cardiotoxicity by affecting amino acid and glucose metabolism.
Aconitine/analogs & derivatives* ; Animals ; Biomarkers/metabolism* ; Chromatography, High Pressure Liquid ; Metabolome ; Metabolomics ; Rats

Aim Air-liquid interface (ALI) cultures of mouse tracheal epithelial cells (MTEC) are a well-established model to study airway epithelial cells. MTEC provides a powerful ap¬proach for the evaluation of the inhalation toxicological in vitro. Methods C57BL/6 mouse tracheal-bronchial epithelial cells were obtained by digestion with protease in cold temperature o- vemight, and the digestion time was optimized to ensure the quantity and viability of the obtained cells. The cells were cul¬tured into collagen coated Transwell inserts. Proliferating phase and air-liquid interface culture were promoted with different cul¬ture media. The expression of tight junction protein and cell trans-epithelial electrical resistance(TEER) were used to evalu¬ate the formation of tight junction between cells and the analysis of cell polarity. The cilia structure was confirmed by electron mi¬ croscopy and immunofluorescence. Results Highly purified and viable primary airway epithelial cells could be harvested and subcultured by our methods, including morphology and immuno- cytochemistry staining confirmed the expression of MUC5AC, a- tubulin, p-tubulin-IV and ZO-1. The development of tight-junc¬tions and epithelium were similar with pseudostratified ciliated columnar epithelium morphology. Conclusions A comprehen¬sive protocol for ALI culture was established, reproducing the characteristic pseudostratified ciliated columnar epithelium mor¬phology and physiological functions in vitro. The MTEC protocol provides a stable and reliable method for the isolation, mucocili¬ary differentiation and reproducing.

OBJECTIVE: Increased carotid artery intima-media thickness (CIMT) and carotid plaque as manifestations of carotid atherosclerosis have been used as markers of cardiovascular disease (CVD). The components of metabolic syndrome (MetS) are linked to CVD, but the association between MetS and CVD is controversial. METHODS: A total of 8,933 Chinese adults aged 40 years or older from 2010 to 2014 were selected from the Jidong and Kailuan communities. MetS was defined by the International Diabetes Federation criteria. CIMT and carotid plaque were measured using color Doppler ultrasound. Logistic regression models were used to assess the association of MetS with carotid plaque and CIMT. RESULTS: MetS was found among 3,461 (3,461/8,933) participants. The odds ratio and 95% confidence internal (CI) for carotid plaques in participants with MetS was 1.16 (1.03-1.30). The risk of carotid plaques increased with the number of MetS components. The average CIMT was higher in participants with MetS (β = 0.020, 95% CI, 0.014-0.027) and in participants with more MetS components. CONCLUSION Individuals with MetS are at an increased risk for carotid atherosclerosis compared to those without MetS.

OBJECTIVE: To explore the role of Ca-NFAT signaling pathway in Ph-ALL drug resistance mediated by bone marrow stromal cells. METHODS: The transcription level of NFAT mRNA in Sup-B15 cells and Ph ALL primary cells was detected by polymerase chain reaction. The expression of P-glycoprotein in Sup-B15 cells was detected by flow cytometry. The change of NFAT protein in Sup-B15 cells was detected by Western blot. AnnexinV/7-AAD was used to label cells. Flow cytometry was used to detect cell apoptosis; Fluo 3-AM dye was used to label cells, and flow cytometry used to detect changes of Ca concentration in leukemia cells. RESULTS NFAT expression could be detected in both Sup-B15 and Ph ALL primary cells; P-glycoprotein could not be detected by flow cytometry; CAS could significantly inhibit NFAT protein expression in clinically applied drug concentrations (2.5, 5 μmol/L); Clinically applied concentration of CAS (2.5, 5 μmol / L) has no significant effect on the apoptosis of Sup-B15 cells, while higher concentration of CAS (10 μmol / L) could induce apoptosis of Sup-B15 cells. Bone marrow stromal cells OP9 could, decrease the sensitivity of Sup-B15 cells and Ph ALL primary cells to imatinib (IM); After co-culture with bone were marrow stromal cells, the Ca concentration in Sup-B15 cells was enhanced, the levels of NFAT protein and nullear protein in sup-B15 cells also were enhanced. The addition of CAS in co-culture system could inlibit the Ca-NFAT signaling pathway, reduce the protective effect of OP9 on Sup-B15 cells.Conclution:The Ca-NFAT sigualing pathway, contributes to the survival of Ph ALL cells. Bone marrow stromal cells can mediate the resistance of Ph ALL cells to IM by activating Ca-NFAT signaling pathway.
Bone Marrow Cells ; Cell Line, Tumor ; Humans ; Imatinib Mesylate ; Mesenchymal Stem Cells ; NFATC Transcription Factors ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; Signal Transduction

The present study aims to investigate the lignans from the flower buds of Magnolia biondii. The isolation and purification of the compounds were performed by column chromatographies on Diaion HP-20, silica gel, and Sephadex LH-20, combined with semi-preparative HPLC. Their structures were elucidated on the basis of spectral data and physiochemical properties. Eighteen compounds were isolated and identified as magnolin (1), epimagnolin (2), eudesmin (3), kobusin (4), aschantin (5), lirioresinol B dimethyl ether (6), pinoresinol monomethy ether (7), (+)-de-O-methylmagnolin (8), isoeucommin A (9), syringaresinol 4-O-β-D-glucopyranoside (10), phillygenin (11), lariciresinol-4'-O-β-1-D-glucoside (12), conicaoside (13), (7'S, 8'R)-dihydrodehydrodiconiferylalcohol (14), 7R*, 8S*-dihydrodehydrodiconiferyl alcohol 4-O-β-D-glucopyranoside (15), 7S, 8R-erythro-7, 9, 9'-trihydroxy-3, 3'-dimethoxy-8-O-4'-neolignan 4-O-β-D-glucopyranoside (16), 7S, 8R-erythro-4, 9, 9'-trihydroxy-3, 3'-dimethoxy-8-O-4'-neolignan-7-O-β-D-glucopyranoside (17), and (+)-isolariciresinol (18). Compounds 7-18 are isolated from this plant for the first time.

Objective To explore the efficacy of ganoderma lucidum preparation(Ling Zhi) in treating APP/PS-1 transgenic mouse models of Alzheimer's disease(AD).Methods APP/PS-1 transgenic mice of 4 months were randomly divided into model group,ganoderma lucidum treatment groups,including high [2250 mg/(kg·d)] and middle [750 mg/(kg·d)] dose groups,i.e.LZ-H and LZ-M groups,and the positive control group(treated with donepezil hydrochloride [2 mg/(kg·d)]).In addition,C57BL/6J wild mice were selected as normal group.The animals were administered for 4 months.Histopathological examinations including hematoxylin-eosin(HE) staining,immunohistochemistry,special staining,and electron microscopy were applied,and then the pathological morphology and structures in different groups were compared. Results The senile plaques and neurofibrillar tangles in the cerebrum and cerebellum were dissolved or disappeared in LZ-H and LZ-M groups.Decrease of amyloid angiopathy was found in LZ-H and LZ-M groups.The immature neurons appeared more in hippocampus and dentate nucleus of LZ-H and LZ-M groups than those in AD model and donepezil hydrochloride groups(hippcampus:F=1.738,P=0.016;dentate nucleus:F=1.924,P=0.026),and these immature neurons differentiated to be neurons.More Purkinje cells loss occurred in AD model mice than that in LZ-H and LZ-M groups(F=9.46,P=0.007;F=9.46,P=0.010).The LZ-H and LZ-M groups had more new neuron stem cells grown up in cerebellum.Electromicroscopic examination showed the hippocampal neurons in LZ-H and LZ-M group were integrated,the nuclear membrane was intact,and the mitochondria in the cytoplasm,endoplasmic reticulum,Golgi bodies,microtubules,and synapses were also complete.The microglial cell showed no abnormality.No toxicity appeared in the pathological specimens of mice treated with ganoderma lucidum preparation.Conclusion The ganoderma lucidum preparation can dissolve and decline or dismiss the senile plaques and neurofibrillar tangles in the brain of AD mice and also reduce the amyloid angiopathy.