Soil cadmium pollution can adversely affect the cultivation of the ornamental plant, Coleus scutellarioides. Upon cadmium contamination of the soil, the growth of C. scutellarioides is impeded, and it may even succumb to the toxic accumulation of cadmium. In this study, we investigated the effects of arbuscular mycorrhizal fungi (AMF) on the adaptation of C. scutellarioides to cadmium stress, by measuring the physiological metabolism and the degree of root damage of C. scutellarioides, with Aspergillus oryzae as the test fungi. The results indicated that cadmium stress increased the activity of superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT), and the content of malondialdehyde (MDA) and proline (Pro) within the cells of C. scutellarioides, but inhibited mycorrhizal infestation rate, root vigour and growth rate to a great degree. With the same cadmium concentration, the inoculation of AMF significantly improved the physiological indexes of C. scutellarioides. The maximum decrease of MDA content was 42.16%, and the content of secondary metabolites rosemarinic acid and anthocyanosides could be increased by up to 27.43% and 25.72%, respectively. Meanwhile, the increase of root vigour was as high as 35.35%, and the DNA damage of the root system was obviously repaired. In conclusion, the inoculation of AMF can promote the accumulation of secondary metabolites, alleviate root damage, and enhance the tolerance to cadmium stress in C. scutellarioides.
Cadmium/toxicity* ; Mycorrhizae/physiology* ; Plant Roots/drug effects* ; Soil Pollutants/toxicity* ; Stress, Physiological ; Superoxide Dismutase/metabolism*

Laccases (LACs), belonging to the multicopper oxidase family, are closely associated with various biological functions including lignin synthesis and responses to biotic and abiotic stresses in plants. However, few studies have reported the laccase genes in China rose (Rosa chinensis). Prickles cause difficulties to the management and harvest of R. chinensis and have become a trait concerned in the breeding. To investigate the expression patterns of laccase genes in roses, we cloned a laccase gene from an ancient variety R. chinensis 'Old Blush' and named it RcLAC15. The expression level of RcLAC15 in prickles was significantly higher than those in roots, stems, and leaves. Fifty-eight laccase genes were identified in the genome of R. chinensis, and bioinformatics analysis revealed that RcLAC15 was a homolog of AtLAC15, predicting that RcLAC15 was a stable hydrophilic protein without transmembrane structures. The recombinant expression vector pBI121-proRcLAC15:: GUS was introduced into Arabidopsis, and GUS staining results showed that the RcLAC15 promoter specifically drove GUS gene expression at the edges of Arabidopsis leaves. In summary, RcLAC15 is a gene specifically expressed in the prickles of R. chinensis. This discovery provides a reference for exploring the biological functions of laccase genes in the prickles of R. chinensis.
Laccase/metabolism* ; Rosa/enzymology* ; Cloning, Molecular ; Gene Expression Regulation, Plant ; Plant Proteins/metabolism* ; Arabidopsis/metabolism* ; Plants, Genetically Modified/metabolism*

OBJECTIVE：To study the co rrelation of the chro maticity value of Schizonepeta tenuifolia charcoal of different processing time（0-40 min，similarly herein after）with multiple indicators ，and to reveal the quality change law of S. tenuifolia charcoal during processing and confirm the terminal time. METHODS ：The contents of ethanol-soluble extracts from S. tenuifolia charcoal decoction pieces of different processing time were determined. UPLC fingerprint of S. tenuifolia decoction pieces and S. tenuifolia charcoal decoction pieces of different processing time were established ，and the similarity evaluation was also conducted. The chromatographic peaks were confirmed by comparison with substance control. The same UPLC conditions were used to determine the contents of index components （hesperidin，rosmarinic acid ，menthone）in S. tenuifolia charcoal decoction pieces of different processing time. The colorimetric method was used to measure the chromaticity value of S. tenuifolia charcoal decoction pieces of different processing time. Meanwhile ，sample of processing 0 min was used as a control to calculate the total color value （E）and the total color difference value （ΔE）. Pearson correlation analysis ，cluster analysis and orthogonal partial least squares discriminant analysis （OPLS-DA）were performed on the ethanol-soluble extracts ，index component contents ，chromatographic peak area and chromaticity value. The terminal time of processing S. tenuifolia charcoal was conf irmed，and validation test was also conducted. RESULTS ：With the extension of processing time ， the content of ethanol-soluble extract in S. tenuifolia charcoal qq.com decoction pieces gradually decreased. A total of 17 chromato- graphic peaks were identified in 12 batches of S. tenuifolia decoction piece ，and its si milarity with the control fingerprint was greater than 0.9. 21 chromatographic peaks were identified in S. tenuifolia charcoal decoction pieces of different processing time，and its similarity with the chromatogram of sample of processing 0 min decreased with the processing time ，and the similarity after 18 min was lower than 0.9. The chromatographic peak 9 was hesperidin ，peak 10 was rosmarinic acid and peak 17 was menthone. The determination of content and chromaticity value showed that with the extension of processing time ，the contents of hesperidin ，rosmarinic acid and menthone decreased gradually ；the color L，b and E values of S. tenuifolia charcoal decoction piece powder decreased gradually ，and the a and ΔE values increased gradually. Pearson correlation analysis showed that the contents of ethanol-soluble extract ，hesperidin，rosmarinic acid and menthone ，the peak areas of 15 chromatographic peaks （peak 2，7-15，17-21）were significantly positively correlated with E value（P＜0.01）；the peak areas of 5 chromatographic peaks （peak 1，3-6）were significantly negatively correlated with E value（P＜0.01），but peak area of peak 16 was not related to E value（P＞ 0.05）. Results of cluster analysis showed that S. tenuifolia charcoal decoction pieces of different processing time were divided into 2 categories；the first category was processed for 0-16 min，and the second category was processed for 18-40 min. The results of OPLS-DA showed that the VIP values of peak 6 area（2.800 75），L value（2.327 54），peak 3 area（1.793 39），b value（1.735 78） and peak 5 area（1.244 04）were greater than 1. The final processing time of S. tenuifolia charcoal was 18 min. The results of validation experiment showed that the L，a and b values of S. tenuifolia charcoal decoction piece were 20.22-22.00，4.44-7.67， 9.78-13.00，and ΔE were 13.50-14.12，respectively. CONCLUSIONS ：The chromaticity value of S. tenuifolia charcoal decoction pieces of different processing time is closely related to the contents of ethanol-soluble extract ，hesperidin，rosmarinic acid ， menthone and the area of 20 chromatographic peaks. It is suggested that the terminal time of processing S. tenuifolia is 18 min.

A new coronavirus (SARS-CoV-2) has been identified as the etiologic agent for the COVID-19 outbreak. Currently, effective treatment options remain very limited for this disease; therefore, there is an urgent need to identify new anti-COVID-19 agents. In this study, we screened over 6,000 compounds that included approved drugs, drug candidates in clinical trials, and pharmacologically active compounds to identify leads that target the SARS-CoV-2 papain-like protease (PLpro). Together with main protease (M
Antiviral Agents/therapeutic use* ; Binding Sites ; COVID-19/virology* ; Coronavirus Papain-Like Proteases/metabolism* ; Crystallography, X-Ray ; Drug Evaluation, Preclinical ; Drug Repositioning ; High-Throughput Screening Assays/methods* ; Humans ; Imidazoles/therapeutic use* ; Inhibitory Concentration 50 ; Molecular Dynamics Simulation ; Mutagenesis, Site-Directed ; Naphthoquinones/therapeutic use* ; Protease Inhibitors/therapeutic use* ; Protein Structure, Tertiary ; Recombinant Proteins/isolation & purification* ; SARS-CoV-2/isolation & purification*

Objective: A method for identification of root cortex and woody core of Morindae Officinalis Radix (MOR) was established based on Ultra Performance Liquid Chromatography (UPLC) characteristic chromatogram and chemical pattern recognition technique. Methods: Using UPLC technique, the characteristic chromatograms of iridoids and oligosaccharides of root cortex and woody core of MOR were established, combined with the similarity analysis, variance analysis, cluster analysis, principal component analysis (PCA) methods for chemical pattern recognition research. Results: The UPLC characteristic chromatograms of iridoids and oligosaccharides of different parts of MOR were established, and 12 and 20 common characteristic peaks were confirmed, respectively. The UPLC characteristic chromatograms of root cortex and woody core of MOR were obviously different. Conclusion: UPLC characteristic chromatograms of iridoids and oligosaccharides of MOR combined with chemical pattern recognition analysis method can reflect the difference of root cortex and woody core of MOR integrally, comprehensively and truly, which provides more sufficient basis for the necessity of removing woody core from MOR.

PURPOSE: Staphylococcal enterotoxin B (SEB) has been well-documented to induce liver injury. miRNA-222-3p (miR-222-3p) was implicated in SEB-induced lung injury and several liver injuries. This study aimed to explore the role of miR-222-3p in SEB-induced liver injury. MATERIALS AND METHODS: Expression of miR-222-3p and suppressors of cytokine signaling 1 (SOCS1) was detected using real-time quantitative PCR and western blot. Liver injury was determined by levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), and inflammatory cytokines, numbers of infiltrating mononuclear cells using AST/ALT assay kit, enzyme-linked immunosorbent assay (ELISA), and hematoxylin-eosin staining, respectively. Target binding between miR-222-3p and SOCS1 was predicted on targetScan software, and confirmed by luciferase reporter assay. RESULTS: SEB induced liver injury in D-galactosamine (D-gal)-sensitized mice, as demonstrated by increased serum levels of AST and ALT, elevated release of interferon-gamma (INF-γ), tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), and IL-2, and promoted infiltrating immune cells into liver. Expression of miR-222-3p was dramatically upregulated, and SOCS1 was downregulated in SEB-induced liver injury both in mice and splenocytes. Moreover, miR-222-3p knockout (KO) mice exhibited alleviated liver injury accompanied with SOCS1 upregulation. Besides, splenocytes under SEB challenge released less INF-γ, TNF-α, IL-6, and IL-2 during miR-222-3p knockdown. Mechanically, SOCS1 was targeted and downregulated by miR-222-3p. Upregulation of SOCS1 attenuated INF-γ, TNF-α, IL-6, and IL-2 release in SEB-induced splenocytes; downregulation of SOCS1 could block the suppressive role of miR-222-3p knockdown in SEB-induced splenocytes. CONCLUSION: Inhibition of miR-222-3p relieves SEB-induced liver inflammatory injury by upregulating SOCS1, thereby providing the first evidence of miR-222-3p in SEB-induced liver injury.
Alanine Transaminase ; Animals ; Aspartate Aminotransferases ; Blotting, Western ; Cytokines ; Down-Regulation ; Enterotoxins ; Enzyme-Linked Immunosorbent Assay ; Interferon-gamma ; Interleukin-2 ; Interleukin-6 ; Liver ; Luciferases ; Lung Injury ; Mice ; Polymerase Chain Reaction ; Tumor Necrosis Factor-alpha ; Up-Regulation

Pathology is a subject that studies the etiology,pathogenesis,pathological changes,progression and outcome of diseases.Pathology links the basic research and clinical practice and is an important part of translational medicine.In order to cultivate qualified pathological graduates with solid pathological theories and the abilities of proposing and addressing scientific hypotheses from pathological morphology changes,we employ modularized special training to divide the pathology training courses into morphology learning module,article searching and reading module,project design module,experiment operation module and scientific presentation module.The training contents among these modules are relatively independent but closely connected,and compose a strategy that aims to improve the scientific innovation ability of pathological graduates.

Objective In our previous study, we had generated various zebrafish mutant lines with tissue?specific GFP expression by Tol2 transposon?mediated insertional mutagenesis. Among these mutants,the Tol2:20141221t line ex?presses GFP in nervous system, while the position within zebrafish genome where transposon inserted has not yet been iden?tified. The aim of this study was to identify and analyze this genetically modified mutant. Methods The transgenic inser?tion loci in the genome of Tol2:20141221t line was identified byTAIL?PCR and the spatial and temporal expression profile of the affected gene was examined by in situ hybridization. Homozygous mutant of Tol2:20141221t was generated for explo?ring related developmental defects. Results Tol2 transposon was inserted into the 8th intron region of sat1.a gene, and in?duced premature transcription termination. The maternal and zygotic mutants of Tol2:20141221t was generated, while with?out apparent developmental defects. Conclusions We have generated and identified the zebrafishsat1.a mutant mediated by Tol2 transposon. This gene insertion mutant exhibits no obvious developmental abnormalities, but may serve as a power?ful tool to study the development of nervous system.

Objective To clarify the influence on component and pharmacological action of Astragalus polysaccharides (APS) as complementary therapeutic agents prepared by different extraction and purification techniques. Methods Components of APS prepared by different extraction and purification techniques were analyzed, and these APS were used for synergy and attenuation of chemotherapy, radiotherapy treatment with H22 liver cancer and Lewis lung cancer of tumor-bearing mice, and also used for the regulation of immune function to immunosuppression mice. Results Experimental data were analyzed by means of statistical method to get pharmaco-result: A3 (extracted by microwave assistance and purified by membrane separation) > A4 (extracted by refluxing and purified by membrane separation) > A1 (extracted by refluxing and no purification)≈ A2 (extracted by microwave assistance and no purification). There were no significant differences on pharmacodynamic action between A1 and A2. However, compared with A1 and A2,it was worth noting that A3 and A4 exhibited good pharmacodynamic action. Then A3-in and A4-in, the samples in dialyzer after dialysis, were separated and purified to get homogeneous APS, which were the principal constituents of APS in dialyzer, with the molecular weight (Mw) of 7669 and 14 142 determined by HPGPC, respectively. The average Mw of APS outside of the dialyzer, A3-out was 3102 and A4-out 3256, which were the main compositions of A3 and A4, accounted for 79.63% and 53.92%, respectively. Conclusion APS with Mw about 5000 Da exhibit better antitumor effect and immunological activity. Refluxing, microwave assistance extractions, and membrane enrichment techniques bring different cases on Mw distribution, components and pharmacodynamic action, and obviously exhibit relationship among component, Mw distribution, and pharmacological action.

Objective To explore an ideal form of graft for pituitary transplantation, which could restore normal pituitary function and avoid hyperprolactinmia providing it is implanted outside the hypophysiotrophic area.Methods Pituitary cells, hypothalamic cells and nigral cells were obtained from human embryos (4-6 months gestation) during therapeutic abortion, and three different cultures were conducted: group P (n=38), half amount of pituitary cells per embryo; group PH (n=18), half amount of pituitary cells mixed with double amount of hypothalamic cells obtained from each embryo; and group PHN (n=20), half amount of pituitary cells mixed with double amount of hypothalamic and nigral cells acquired from each embryo. The histological changes of the cultured cells were observed and the secretive levels of growth hormone and prolactin indifferent medium were measured.Results All the monolayer cultured cells grew well throughout the 30-day culture period. Ultrastructurally, the cells in group P degenerated significantly at the 21st day of culture. However, in the PH and PHN groups, the cells kept well viability. The secretive level of growth hormone in culture medium declined steadily in group P, but kept at 20 ng/ml in both PH and PHN groups. Prolactin concentration decreased rapidly following an initial increase in group P, but kept at a high level in PH group. In PHN group, the prolactin level was valuable, remaining at about 15 ng/ml.Conclusions In the mixed culture of fetal pituitary-hypothalamic-nigral cells, the pituitary cells kept high viability in a long period and its hormone secretion remained at ideal levels. These indicate that the mixed fetal pituitary-hypothalamic-nigral cells are a good graft, which can be implanted outside the cranium while maintaining normal pituitary function with no increase in blood prolactin concentration.