Artemisia argyi is a perennial herbaceous herb of the Artemisia family, with leaves for medical use. However, the germplasm of A. argyi is seriously unclear and mixed during production, and it is urgent to protect new varieties of A. argyi. The distinctness, uniformity, and stability(DUS) testing of the new varieties of plants is the basis for the protection of new varieties of plants, and the development of the DUS testing guidelines is the technical basis for DUS testing. To develop the DUS testing guidelines of A. argyi, A. argyi of 100 germplasm was used as the research objects, and their agronomic and medicinal quality characters were observed and measured during six growth stages, and each character was graded and described. A total of 53 test characters were determined, including 19 characters that must be tested; there were four plant characters, two rhizome characters, five stem characters, three branching characters, 29 leaf characters, three floral characters, five medicinal quality characters, and two other characters. It also involved 16 quality characters, 22 quantitative characters, and 15 pseudo-quantitative characters. Seven grouping characters were determined from 53 characters, including "emergence period" "plant-plant type" "branching-primary branching site" "stem-color" "middle leaf-number of leaf splits" "budding period", and "plant-height". By searching for standard characters, 16 standard varieties were ultimately determined. The preparation of this guideline was of great significance for the review and protection of new A. argyi varieties, the protection of breeders' rights, and the promotion of the development of A. argyi industry.
Artemisia/chemistry* ; Plant Leaves/chemistry* ; Quality Control ; Plants, Medicinal/classification* ; Guidelines as Topic

Lithocarpus litseifolius is rich in the chalcones phloridzin and trilobatin, the biosynthesis pathways of which have not been fully demonstrated. Chalcone synthase(CHS) is the first key rate-limiting enzyme in the biosynthesis of flavonoids in plants. To explore the functions of CHS gene family in chalcone synthesis of L. litseifolius, this study screened out two CHS genes(LlCHS1 and LlCHS2) from the transcriptome data of this plant, and then bioinformatics analysis and functional characterization were performed for the two genes. The bioinformatics analysis showed that LlCHS1 and LlCHS2 were acidic hydrophilic stable proteins with no transmembrane domain, composed of 395 and 390 amino acid residues, respectively. Both of them contained the characteristic amino acid sequence "WGVLFGFGPGL" and highly conserved active sites(Cys-164, Phe-215, His-303, and Asn-336) of the CHS family. The phylogenetic tree showed that LlCHS1 shared the same clade with similar genes in Aquilaria sinensis, and LlCHS2 was closely related to similar genes in Malus domestica. Under exogenous addition of phloretic acid, co-expression of LlCHS1 or LlCHS2 with Aa4CL from Aromatoleum aromaticum in Escherichia coli catalyzed the production of phloretin from phloretic acid. This study laid a theoretical foundation for revealing the functions of CHS in plants and provided new enzymatic modules for producing phloretin by synthetic biology.
Acyltransferases/chemistry* ; Phylogeny ; Plant Proteins/chemistry* ; Cloning, Molecular ; Amino Acid Sequence

This study aimed to explore the anti-inflammatory material basis and molecular mechanism of Artemisia stolonifera based on the analysis of the chemical components in different extracted fractions of A. stolonifera and their antioxidant and anti-inflammatory effects in combination with network pharmacology and molecular docking. Thirty-two chemical components were identified from A. stolonifera by ultra-performance liquid chromatography coupled to tandem quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS). Among them, there were 7, 21 and 22 compounds in water, n-butanol and ethyl acetate fractions, respectively. The antio-xidant capacity of different extracted fractions was evaluated by measuring their scavenging ability against 1,1-diphenyl-2-picrylhydrazyl radical 2,2-diphenyl-1-(2,4,6-trinitrophenyl) hydrazyl(DPPH) and 2,2'-azinobis-(3-ethylbenzthiazoline-6-sulphonic acid)(ABTS) free radicals and total antioxidant capacity [ferric reducing antioxidant power(FRAP) assay]. The inflammatory model of RAW264.7 cells was induced by lipopolysaccharide(LPS), and the levels of nitrite oxide(NO), tumor necrosis factor-α(TNF-α), interleukin-6(IL-6) in the supernatant and the mRNA expression of related inflammatory factors in cells were used to evaluate the anti-inflammatory effects. The results revealed that ethyl acetate fraction of A. stolonifera was the optimal antioxidant and anti-inflammatory fraction. By network pharmacology, it was found that flavonoids such as rhamnazin, eupatilin, jaceosidin, luteolin and nepetin could act on key targets such as TNF, serine/threonine protein kinase 1(AKT1), tumor protein p53(TP53), caspase-3(CASP3) and epidermal growth factor receptor(EGFR), and regulate the phosphatidylinositol-3-kinase-protein kinase B(PI3K-AKT) and mitogen-activated protein kinase(MAPK) signaling pathways to exert the anti-inflammatory effects. Molecular docking further indicated excellent binding properties between the above core components and core targets. This study preliminarily clarified the anti-inflammatory material basis and mechanism of ethyl acetate fraction of A. stolonifera, providing a basis for the follow-up clinical application of A. stolonifera and drug development.
Antioxidants/chemistry* ; Molecular Docking Simulation ; Artemisia ; Network Pharmacology ; Phosphatidylinositol 3-Kinases ; Anti-Inflammatory Agents/chemistry* ; Drugs, Chinese Herbal/pharmacology* ; Interleukin-6

OBJECTIVE: To evaluate the effects of electromyography on the clinical manifestations and prognosis after posterior lumbar interbody fusion(PLIF) of degenerative lumbar diseases. METHODS: A retrospective analysis was performed on 68 patients with degenerative lumbar diseases, including 29 males and 39 females, aged 21 to 84 years old, who underwent electromyogram (EMG) from January 2018 to October 2019. The patients were divided into negative and positive groups according to whether theresults of EMG was normal or abnormal, PLIF surgery was performed in both groups. The preoperative duration of illness, postoperative recovery time, operative time, intraoperative blood loss, postoperative ambulation time and length of postoperative hospital stay were recorded. The clinical efficacy was evaluated by visual analogue scale(VAS) of low back and lower limb, the Japanese Orthopedic Association(JOA) score before and after operation. RESULTS: All patients were follow-up from 26 to 39 months. The subjective symptoms, clinical signs, daily activities and JOA total scores after operation in two groups were significantly higher than those before preoperation(P<0.05);the clinical signs score and total JOA score in the negative group at 3 months after operation were higher than those in the positive group(P<0.05). The VAS score of leg pain in the negative group after 1 and 3 months was less than that in the positive group(P<0.05). Patients 's illness time, postoperative recovery time, hospitalization time and implantation time in the negative group were shorter than those in the positive group(P<0.05). At other time points, there was no significant difference in low pain VAS, leg pain VAS, JOA scores in the two groups(P>0.05). There was no significant difference in the operation time and intraoperative bleeding volume between the two groups(P>0.05). CONCLUSION Patients with normal electromyography had shorter disease duration than ones with abnormal electromyography in lumbar degenerative disease;after PLIF, patients with normal electromyography recovered faster than ones with abnormal electromyography, but the results of electromyography had no effect on the final prognosis of PLIF surgery.
Male ; Female ; Humans ; Young Adult ; Adult ; Middle Aged ; Aged ; Aged, 80 and over ; Retrospective Studies ; Spinal Fusion/methods* ; Lumbar Vertebrae/surgery* ; Treatment Outcome ; Pain

This study explores the effect of apigenin(APG), oxymatrine(OMT), and APG+OMT on the proliferation of non-small cell lung cancer cell lines and the underlying mechanisms. Cell counting kit-8(CCK-8) assay was used to detect the vitality of A549 and NCI-H1975 cells, and colony formation assay to evaluate the colony formation ability of the cells. EdU assay was employed to examine the proliferation of NCI-H1975 cells. RT-qPCR and Western blot were performed to detect the mRNA and protein expression of PLOD2. Molecular docking was carried out to explore the direct action ability and action sites between APG/OMT and PLOD2/EGFR. Western blot was used to study the expression of related proteins in EGFR pathway. The viability of A549 and NCI-H1975 cells was inhibited by APG and APG+OMT at 20, 40, and 80 μmol·L~(-1) in a dose-dependent manner. The colony formation ability of NCI-H1975 cells was significantly suppressed by APG and APG+OMT. The mRNA and protein expression of PLOD2 was significantly inhibited by APG and APG+OMT. In addition, APG and OMT had strong binding activity with PLOD2 and EGFR. In APG and APG+OMT groups, the expression of EGFR and proteins in its downstream signaling pathways was significantly down-regulated. It is concluded that APG in combination with OMT could inhibit non-small lung cancer, and the mechanism may be related to EGFR and its downstream signaling pathways. This study lays a new theoretical basis for the clinical treatment of non-small cell lung cancer with APG in combination with OMT and provides a reference for further research on the anti-tumor mechanism of APG in combination with OMT.
Humans ; Carcinoma, Non-Small-Cell Lung ; Lung Neoplasms ; Apigenin ; Molecular Docking Simulation ; Alkaloids ; Quinolizines ; RNA, Messenger ; ErbB Receptors

To clarify the content characteristics of mineral elements in different Artemisia argyi germplasm resources and their relationship with the quality properties of Artemisiae Argyi Folium, this study measured the content of 10 mineral elements including nitrogen(N), phosphorus(P), potassium(K), calcium(Ca), magnesium(Mg), aluminum(Al), manganese(Mn), iron(Fe), copper(Cu), and zinc(Zn) in 100 Artemisia argyi germplasm samples. Besides, their relationship with the quality properties of Artemisiae Argyi Folium was explored by correlation analysis, path analysis, and cluster analysis. The results demonstrated that the variation coefficient of the 10 mineral elements in Artemisiae Argyi Folium ranged from 12.23% to 64.38%, and the genetic diversity index from 0.97 to 3.09. The genetic diversities of N, P, and Zn were obvious. As revealed by the correlation analysis, N, P, and K showed strong positive correlations with each other. Except that Mg and Al were negatively correlated, Ca, Mg, Al, Mn, Fe, Cu, and Zn were positively correlated. The correlation analysis of mineral elements with the quality properties of Artemisiae Argyi Folium proved the significant correlations of 17 pairs of characters. According to the path analysis, P, K, Ca, and Mn greatly affected the yield of Artemisiae Argyi Folium, P, K, and Mg the output rate of moxa, N, P, and K the content of total volatile oil, P and K the content of eucalyptol, and P, K, and Ca the content of eupatilin. The 100 germplasm samples were clustered into three groups. Specifically, in cluster Ⅰ, the enrichment capacity of P, K, and Mg elements was strong, and the comprehensive properties of mineral elements were better, implying good development potential. Ca, Mn, Fe, and Zn elements in cluster Ⅱ and N and Al in cluster Ⅲ displayed strong enrichment capacities. This study has provided new ideas for resource evaluation and variety breeding of A. argyi and also reference for fertilizer application.
Artemisia/genetics* ; Iron ; Minerals/analysis* ; Plant Breeding ; Plant Leaves/chemistry*

Objective: To explore the heterogeneity and correlation of clinical phenotypes and genotypes in children with disorders of sex development (DSD). Methods: A retrospective study of 1 235 patients with clinically proposed DSD in 36 pediatric medical institutions across the country from January 2017 to May 2021. After capturing 277 DSD-related candidate genes, second-generation sequencing was performed to analyzed the heterogeneity and correlation combined with clinical phenotypes. Results: Among 1 235 children with clinically proposed DSD, 980 were males and 255 were females of social gender at the time of initial diagnosis with the age ranged from 1 day of age to 17.92 years. A total of 443 children with pathogenic variants were detected through molecular genetic studies, with a positive detection rate of 35.9%. The most common clinical phenotypes were micropenis (455 cases), hypospadias (321 cases), and cryptorchidism (172 cases) and common mutations detected were in SRD5A2 gene (80 cases), AR gene (53 cases) and CYP21A2 gene (44 cases). Among them, the SRD5A2 mutation is the most common in children with simple micropenis and simple hypospadias, while the AMH mutation is the most common in children with simple cryptorchidism. Conclusions: The SRD5A2 mutation is the most common genetic variant in Chinese children with DSD, and micropenis, cryptorchidism, and hypospadias are the most common clinical phenotypes. Molecular diagnosis can provide clues about the biological basis of DSD, and can also guide clinicians to perform specific clinical examinations. Target sequence capture probes and next-generation sequencing technology can provide effective and economical genetic diagnosis for children with DSD.
3-Oxo-5-alpha-Steroid 4-Dehydrogenase/genetics* ; Child ; China/epidemiology* ; Cryptorchidism/genetics* ; Disorders of Sex Development/genetics* ; Female ; Genital Diseases, Male ; Genotype ; Humans ; Hypospadias/genetics* ; Male ; Membrane Proteins/genetics* ; Penis/abnormalities* ; Phenotype ; Retrospective Studies ; Steroid 21-Hydroxylase/genetics*

Background: and Purpose Recent studies suggested an increased incidence of cerebral venous thrombosis (CVT) during the coronavirus disease 2019 (COVID-19) pandemic. We evaluated the volume of CVT hospitalization and in-hospital mortality during the 1st year of the COVID-19 pandemic compared to the preceding year. Methods: We conducted a cross-sectional retrospective study of 171 stroke centers from 49 countries. We recorded COVID-19 admission volumes, CVT hospitalization, and CVT in-hospital mortality from January 1, 2019, to May 31, 2021. CVT diagnoses were identified by International Classification of Disease-10 (ICD-10) codes or stroke databases. We additionally sought to compare the same metrics in the first 5 months of 2021 compared to the corresponding months in 2019 and 2020 (ClinicalTrials.gov Identifier: NCT04934020). Results: There were 2,313 CVT admissions across the 1-year pre-pandemic (2019) and pandemic year (2020); no differences in CVT volume or CVT mortality were observed. During the first 5 months of 2021, there was an increase in CVT volumes compared to 2019 (27.5%; 95% confidence interval [CI], 24.2 to 32.0; P<0.0001) and 2020 (41.4%; 95% CI, 37.0 to 46.0; P<0.0001). A COVID-19 diagnosis was present in 7.6% (132/1,738) of CVT hospitalizations. CVT was present in 0.04% (103/292,080) of COVID-19 hospitalizations. During the first pandemic year, CVT mortality was higher in patients who were COVID positive compared to COVID negative patients (8/53 [15.0%] vs. 41/910 [4.5%], P=0.004). There was an increase in CVT mortality during the first 5 months of pandemic years 2020 and 2021 compared to the first 5 months of the pre-pandemic year 2019 (2019 vs. 2020: 2.26% vs. 4.74%, P=0.05; 2019 vs. 2021: 2.26% vs. 4.99%, P=0.03). In the first 5 months of 2021, there were 26 cases of vaccine-induced immune thrombotic thrombocytopenia (VITT), resulting in six deaths. Conclusions During the 1st year of the COVID-19 pandemic, CVT hospitalization volume and CVT in-hospital mortality did not change compared to the prior year. COVID-19 diagnosis was associated with higher CVT in-hospital mortality. During the first 5 months of 2021, there was an increase in CVT hospitalization volume and increase in CVT-related mortality, partially attributable to VITT.

Objective:To explore the effect of different proportions of organic fertilizer and chemical fertilizer on the agronomic traits， yield and quality of Artemisiae Argyi Folium in Qichun county， and provide a theoretical basis for scientific fertilization of its planting. Method:Field plot experiment was carried out to set up 5 treatment methods with different proportions of organic fertilizers and chemical fertilizers［OM₀ （no combined application of biological organic fertilizer）， OM₁₇ （combined application of 17% biological organic fertilizer）， OM₃₃ （combined application of 33% biological organic fertilizer）， OM₆₇ （combined application of 67% biological organic fertilizer）， OM₁₀₀ （combined application of 100% biological organic fertilizer）］. The effects of different treatment methods on the agronomic characters， leaf yield， output rate of moxa， volatile oil content， flavonoid and phenolic acid contents and mineral element contents of Artemisiae Argyi Folium in Qichun county were determined. Result:With the increase of the proportion of organic fertilizer in application， the seedling number per unit area， plant height， stem diameter， leaf number， leaf width， leaf length， height of dead leaves and leaf yield of Artemisiae Argyi Folium were increased at first and then decreased. Among them， the yield of Artemisiae Argyi Folium in OM₃₃ treatment was 61.37% higher than that in OM₀ treatment. With the increase of the proportion of organic fertilizer， the output rate of moxa of Artemisiae Argyi Folium showed continuously increasing trend， contents of volatile oil and volatile components （eucalyptol， α-thujone， borneol， camphor and caryophyllene oxide） increased at first and then decreased， while the contents of α-caryophyllene and β-syringene decreased gradually， the contents of phenolic acids （neochlorogenic acid， chlorogenic acid， isochlorogenic acid B， A and C） increased at first and then decreased， while the contents of flavonoids （jaceosidin and eupatilin） increased continuously， and the contents of mineral elements （Ca， Cu and Zn） continued to increase， but the content of K decreased significantly at the high proportion of organic fertilizer. After treated with principal component analysis （PCA）， it was found that OM₁₇ treatment had the highest quality， while OM₁₀₀ and OM₀ treatment had low quality. Conclusion:Based on comprehensive analysis of agronomic traits， yield and quality indexes of Artemisiae Argyi Folium in Qichun county， it is suggested that 17%-33% proportion of organic fertilizer should be used in its production， in order to improve the quality and efficiency of Artemisiae Argyi Folium industry in Qichun county.

Objective:To analyze the quality differences among different germplasm resources of Artemisia argyi and to screen out the specific germplasm by comprehensively evaluating 14 quality traits of 100 germplasm resources. Method:Germplasm resources of A. argyi were collected from all over the country. The output rate of moxa and the content of total volatile oil in A. argyi leaves were determined，and the contents of 12 flavonoids and phenolic acids in A. argyi leaves were detected by ultra performance liquid chromatography（UPLC）. The correlation analysis，principal component analysis and clustering analysis were used to comprehensively evaluate the quality of A. argyi. Result:There was rich genetic diversity of A. argyi germplasm resources，and the variation coefficients of 14 quality traits ranged from 25.67% to 127.34%，among which the coefficient of variation of chlorogenic acid，cryptochlorogenic acid，isoxiafotaside，isochlorogenic acid B and isochlorogenic acid A was more than 70%，with high variation. The output rate of moxa was negatively correlated with 9 quality traits，while the content of total volatile oil was positively correlated with 10 quality traits，and most of the flavonoids and phenolic acids had synergistic effects. 12 flavonoids and phenolic acids were analyzed by principal component analysis，and 4 principal components could be extracted. The highest contents of flavonoids and phenolic acids were found in S98（Hangzhou，Zhejiang province），S84（Longhui county，Shaoyang city，Hunan province），S66（Futian river town，Macheng city，Hubei province），S35 （Balihu town，Qichun county，Huanggang city，Hubei province），and S15 （Fudao town，Tangyin county，Anyang city，Henan province）. The systematic clustering analysis showed that the 100 germplasm could be divided into four groups when the euclidean distance was 8.0，with 90，3，3，3 and 4 accessions in group Ⅰ，Ⅱ，Ⅲ and Ⅳ，respectively. The germplasm resources in group Ⅱ contained the highest content of flavonoids and phenolic acids，the group Ⅲ contained the highest content of total volatile oil and the group Ⅳ contained the highest output rate of moxa. Conclusion:The leaf quality of different A. argyi germplasms is different. This study can provide the basis for the quality evaluation and variety breeding of A. argyi germplasm resources.