OBJECTIVE To analyze the serum chemical composition of rats after intragastric administration of water extract of crude Anemarrhenae Rhizoma-crude Phellodendri Chinensis and salted Anemarrhenae Rhizoma-salted Phellodendri Chinensis based on liquid chromatography-mass spectrometry(LC-MS)technology,predict the effect of salt processing on the treatment of type 2 dia-betes in Anemarrhenae Rhizoma-Pheellodendri Chinensis combined with network pharmacology,and preliminarily verify it through in vitro experiments.METHODS Rats were continuously intragastrically administered with crude Anemarrhenae Rhizoma-crude Phello-dendri Chinensis drug pair and salted Anemarrhenae Rhizoma-salted Phellodendri Chinensis drug pair water extract twice,with an in-terval of 1 h.After 60 min of the last administration,the blood was taken from the abdominal aorta,and the protein was precipitated by methanol.After dissolution,the chromatographic column was Shim-pack GIST C18(4.6 mm×150 mm,5 μm);the mobile phase A was 0.1%formic acid water,and the mobile phase B was 0.1%formic acid-acetonitrile;gradient elution,positive and negative ion full scan mode,mass scan range 100-1 500 m/z.Combined with the secondary spectrum of the database and literature,the blood compo-nents of crude Anemarrhenae Rhizoma-crude Phellodendri Chinensis drug pair and salted Anemarrhenae Rhizoma-salted Phellodendri Chinensis drug pair were analyzed and identified.The disease targets of type 2 diabetes were retrieved,and the protein interaction net-work analysis,GO and KEGG pathway enrichment analysis were performed on the intersection targets of blood components and diseases.The"blood components-targets"network diagram was constructed,and the selected core components and core targets were verified by molecular docking using AutoDock software.In the verification experiment,HepG2 cells were used as the experimental ob-ject,and the insulin resistance model was induced by high insulin and high glucose.CCK8 method was used to test the effect of Rhizo-ma Anemarrhenae-Phellodendri Chinensis on cell proliferation before and after salt processing.Western blot was used to detect the ex-pression of PI3K-AKT signaling pathway-related proteins.RESULTS 15 prototype components and 1 mangiferin metabolic compo-nent were identified in the serum of rats.17 prototype components and 1 mangiferin metabolite were identified in the rat serum of the water extract of Anemarrhenae Rhizoma-Phellodendri Chinensis after salt processing.The contents of mangiferin,berberine and 3-isobutylglutaric acid in the blood components after salt-processing were higher than those in the raw products.According to the results of KEGG and GO,the treatment of type 2 diabetes may be related to the transcriptional regulation of RNA polymerase,inflammatory re-sponse,AGE-RAGE,PI3K-AKT pathway and insulin resistance.Cell experiments showed that the ratio of p-PI3K/PI3K,p-AKT/AKT and GLUT4 protein expression could be up-regulated before and after salt processing,and the effect of salt processing group was better than that of the crude group.CONCLUSION This experiment preliminarily revealed the components of Anemarrhenae Rhizo-ma-Phellodendri Chinensis drug pair entering the blood before and after salt exposure,and suggested that ferulic acid,berberine,ber-berrubine,mangiferin,mTOR,SIRT1,EGFR and PPARA may be the main components and targets of Rhizoma Anemarrhenae-Phel-lodendri Chinensis after salt processing to enhance the therapeutic effect of type 2 diabetes.The mechanism may be to enhance the role of PI3K-AKT and other related signaling pathways,providing an important reference for the pharmacodynamic material basis and clini-cal application of Anemarrhenae Rhizoma-Phellodendri Chinensis before and after salt processing.

Aim To investigate how the long non-cod-ing RNA uc.48+(lncRNA uc.48+)affected calci-tonin gene-related peptide(CGRP)in the trigeminal ganglion(TG)of rats with trigeminal neuralgia(TN)and its potential mechanism.Methods Chronic con-striction injury of the infraorbital nerve(CCI-ION)in rats was used to create the animal model for trigeminal neuralgia.After modeling,uc.48+siRNA was injec-ted locally via the infraorbital foramen to knock down lncRNA uc.48+,and uc.48+plasmid was transfect-ed into normal rats to over-express lncRNA uc.48+.The face mechanical pain threshold(MWT)of each group was measured by behavioral test,and the content and changes of CGRP in rat TG were observed using qPCR and protein blotting.The change in serum in-flammatory cytokine 1L-1β was determined using ELISA.Results The MWT in TN rats treated with the uc.48+siRNA increased significantly,but the protein and mRNA levels of CGRP in TG decreased significantly(P＜0.01),and the level of 1L-1β de-creased as well(P＜0.01).In addition,the MWT of normal rats transfected with uc.48+plasmid was sig-nificantly diminished,and the mRNA and protein lev-els of CGRP in TG were markedly elevated(P＜0.01),as were the levels of 1L-1β(P＜0.01),compared to normal rats.Conclusions Knocking out uc.48+in TN rats reduces pain,while overexpressing uc.48+exacerbates pain transmission in trigeminal neuralgia.The mechanism by which uc.48+small in-terference inhibits trigeminal neural pathology pain may be through decreasing CGRP expression in TG of rats with TN,therefore ameliorating mechanical pain sensi-tivity.

Tigliane type macrocyclic diterpenoids with special structures and diverse bioactivities are mainly extracted from plants of Euphorbiaceae and Thymelaeaceae. According to the different functional groups, they can be classified into types of phorbol esters, C-4 deoxyphorbol esters, C-12 deoxyphorbol esters, C-16 or C-17 substituted phorbol esters and others. Most of them present promising antiviral activities and cytotoxic activities and are expected to be developed as candidates for anti-AIDS, anti-tuberculosis, and anti-tumor clinical trials, demonstrating great potential for the application in healthcare. This paper reviews 115 novel tigliane-type diterpenoids discovered since 2013 and summarize their chemical structures and bioactivities, aiming to lay a foundation for further development and utilization of these compounds and provide new ideas for the development of clinical drugs.
Phorbols ; Molecular Structure ; Diterpenes/chemistry* ; Antiviral Agents ; Phorbol Esters

Objective:To analyze the epidemiological characteristics of 8 clusters of coronavirus disease 2019 （COVID-19） in Chenzhou City， and provide scientific basis for epidemic prevention and control. Methods:Descriptive epidemiological analysis was conducted for 8 COVID-19 clusters， comparing and analyzing the differences of infection rates among close contacts within and outside the family， and emphatically describing two typical cases. Results:8 COVID-19 clusters were reported in Chenzhou with a total of 31 cases from January to February， 2020. The main source of infection of the family index cases was Hubei Province. Cough symptoms were observed in 67.74% of the cases， followed by fever （54.84%）. The infection rate of close contacts within the family （55.00%） was higher than that outside the family （2.56%）， and the difference was statistically significant （χ²=28.177， P<0.001）. The infection rate of spouse of the family index cases was 85.71%， higher than that of parents （77.78%）， other family members （44.44%） and children （40.00%）， and the difference was not statistically significant （χ²=6.004， P=0.120）. Two typical cases suggested that both COVID-19 pre-symptomatic and asymptomatic patients have the potential to excrete the virus from the body and become sources of infection. Conclusion:Effective family prevention and control measures and early sampling and screening of people in key epidemic areas are conducive to early detection， early isolation and early treatment of infected people， so as to avoid the occurrence and spread of family clusters.

In the context of the coronavirus disease 2019 (COVID-19)pandemic,thousands of health care wor- kers (HCWs)worldwide infected with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2),some even have lost their lives.At the early stage of the epidemic,some Chinese HCWs were infected.Owing to limited knowledge of characteristics of SARS-CoV-2,more than 3,000 HCWs in Hubei Province contracted SARS-CoV-2 at the early stage of the outbreak.Due to overloaded work of HCWs in local hospitals,more than 42,000 HCWs (including HCWs from the military)were dispatched to Hubei Province from all over the country.At the peak of epidemic,one in 10 intensive care HCWs in China were working in Wuhan.During fighting against COVID-19 in China,although a certain number of HCWs were infected with SARS-CoV-2 at the early stages of the epidemic, effective prevention was achieved through timely adoption of prevention measures,including fast diagnosis,timely isolation of patients,strengthening of HCWs'safety,intensified training on basic protective knowledge and unified management of HCWs,there was no report about infection among the 42,632 members of the national medical teams sent to Hubei,and the number of COVID-19 cases among HCWs in local hospitals also significantly de- creased,thereby indicating that healthcare-associated infection (HAI)of COVID-19 among HCWs are fully pre- ventable.This paper explores how to prevent HCWs from contracting SARS-CoV-2 through effective measures during the epidemic in Wuhan,China.

Plant breeding is well recognized as one of the most important means to meet food security challenges caused by the ever-increasing world population. During the past three decades, plant breeding has been empowered by both new knowledge on trait development and regulation (e.g., functional genomics) and new technologies (e.g., biotechnologies and phenomics). Gene editing, particularly by clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein (Cas) and its variants, has become a powerful technology in plant research and may become a game-changer in plant breeding. Traits are conferred by coding and non-coding genes. From this perspective, we propose different editing strategies for these two types of genes. The activity of an encoded enzyme and its quantity are regulated at transcriptional and post-transcriptional, as well as translational and post-translational, levels. Different strategies are proposed to intervene to generate gene functional variations and consequently phenotype changes. For non-coding genes, trait modification could be achieved by regulating transcription of their own or target genes via gene editing. Also included is a scheme of protoplast editing to make gene editing more applicable in plant breeding. In summary, this review provides breeders with a host of options to translate gene biology into practical breeding strategies, i.e., to use gene editing as a mechanism to commercialize gene biology in plant breeding.

A method was established for simultaneous determination of 21 active constituents including flavanols, isoflavones, flavonols, dihydroflavones, dihydroflavonols, chalcones, pterocarpan, anthocyanidins and phenolic acids in Spatholobi Caulis by ultra fast liquid chromatography with triple quadrupole linear ion trap mass spectrometry(UFLC-QTRAP-MS/MS). Then, it was employed to analyze and evaluate the dynamic accumulation of multiple bioactive constituents in Spatholobi Caulis. The chromatographic separation was performed on a XBridge®C_(18)(4.6 mm×100 mm, 3.5 μm) at 30 ℃ with a gradient elution of 0.3% formic acid aqueous solution-methanol, and the flow rate was 0.8 mL·min~(-1), using multiple-reaction monitoring(MRM) mode. A comprehensive evaluation of the multiple bioactive constituents was carried out by gray correlation analysis(GRA). The 21 target components showed good linearity(r>0.999 0) in the range of the tested concentrations. The average recovery rates of the 21 components were from 97.46% to 103.6% with relative standard deviations less than 5.0%. There were differences in the contents of 21 components in Spatholobi Caulis at diffe-rent harvest periods. Spatholobi Caulis had high quality from early November to early December, which is consistent with the local tradi-tional harvest period. This study reveals the rule of the dynamic accumulation of 21 components in Spatholobi Caulis and provides basic information for the suitable harvest time. At the same time, it provides a new method reference for the comprehensive evaluation of the internal quality of Spatholobi Caulis.
Chromatography, High Pressure Liquid ; Fabaceae/chemistry* ; Phytochemicals/isolation & purification* ; Plant Stems/chemistry* ; Plants, Medicinal/chemistry* ; Tandem Mass Spectrometry

Molecular biology is a new subject that clarifies the phenomena and nature of life at the molecular level. Its development provides new biotechnology and methods for the study of traditional pharmacognosy. The formation of molecular biology has brought the development of pharmacognosy into a new era of gene research. Lonicerae Japonicae Flos is a classical Chinese medicine. Many scholars of home and abroad have carried out relevant studies on its molecular biology on the basis of the in-depth study with traditional methods, and have achieved certain results. In order to provide references on the method, technical for promoting the modernization of Lonicerae Japonicae Flos, and the development, protection, and utilization of other traditional Chinese medicine resources. This article summarized the application status of molecular biology methods and techniques on the identification, biosynthesis of active constituents, and molecular mechanism of secondary metabolite under stress conditions of Lonicerae Japonicae Flos in recent years. In hybridization technology of tag(RFLP), molecular markers based on PCR(RAPD, AFLP, SSR and ISSR), based on DNA sequence analysis of SNP and DNA barcode for the variety identification, diagnosis, identification of Lonicerae Japonicae Flos, and so forth in detail. At the same time, it is proposed that multi-omics technology can be used to build systems biology technology and platforms, and establish related models of secondary metabolite biosynthesis, so as to deepen acknowledge the molecular mechanism of the active component biosynthesis of Lonicerae Japonicae Flos and the accumulation of metabolites, life activities of other medicinal plants under adverse environment, then to regulate them.
Amplified Fragment Length Polymorphism Analysis ; Chromatography, High Pressure Liquid ; DNA Barcoding, Taxonomic ; Drugs, Chinese Herbal/pharmacology* ; Lonicera/chemistry* ; Medicine, Chinese Traditional ; Microsatellite Repeats ; Plants, Medicinal/chemistry* ; Polymorphism, Restriction Fragment Length ; Polymorphism, Single Nucleotide ; Random Amplified Polymorphic DNA Technique ; Secondary Metabolism

BACKGROUND: Desminopathy, a hereditary myofibrillar myopathy, mainly results from the desmin gene (DES) mutations. Desminopathy involves various phenotypes, mainly including different cardiomyopathies, skeletal myopathy, and arrhythmia. Combined with genotype, it helps us precisely diagnose and treat for desminopathy. METHODS: Sanger sequencing was used to characterize DES variation, and then a minigene assay was used to verify the effect of splice-site mutation on pre-mRNA splicing. Phenotypes were analyzed based on clinical characteristics associated with desminopathy. RESULTS: A splicing mutation (c.735+1G>T) in DES was detected in the proband. A minigene assay revealed skipping of the whole exon 3 and transcription of abnormal pre-mRNA lacking 32 codons. Another affected family member who carried the identical mutation, was identified with a novel phenotype of desminopathy, non-compaction of ventricular myocardium. There were 2 different phenotypes varied in cardiomyopathy and skeletal myopathy among the 2 patients, but no significant correlation between genotype and phenotype was identified. CONCLUSIONS We reported a novel phenotype with a splicing mutation in DES, enlarging the spectrum of phenotype in desminopathy. Molecular studies of desminopathy should promote our understanding of its pathogenesis and provide a precise molecular diagnosis of this disorder, facilitating clinical prevention and treatment at an early stage.
Animals ; Asian Continental Ancestry Group ; Cardiomyopathies ; genetics ; pathology ; Desmin ; genetics ; Electrocardiography ; Female ; Genotype ; Humans ; Male ; Middle Aged ; Muscular Dystrophies ; genetics ; pathology ; Mutation ; genetics ; Pedigree ; Phenotype

Objective: To explore the effects of noscapine (Nos) on the expression of cadherin 17 (CDH17) in colon cancer SW480 cells and the mechanism of Nos on cell migration. Methods: SW480 cells were divided into the control group, empty vector (si-EV) group, CDH17 interference (si-CDH17) group, Nos treatment group, and CDH17 interference+Nos treatment (si-CDH17+Nos) group. Small interfering RNA (siRNA) was used to knockdown CDH17, and the selected concentration of Nos was (55.30±2.21) µg/ml (IC50). The mRNA expression of CDH17 was detected by qPCR; the apoptosis and migration abilities of SW480 cells were observed by Hoechst33258 staining and Transwell assay; the contents of VEGF, MMP2 and MMP9 in SW480 cells were measured by ELISA, and the protein expressions of CDH17, Wnt3a and β-catenin were determined by WB. Results: Compared with the control group, mRNA and protein expressions of CDH17 obviously decreased, cell apoptosis and migration significantly reduced, while the contents of VEGF, MMP2 and MMP9 as well as the protein expressions of Wnt3a and β-catenin significantly decreased in Nos treatment group, siCDH17 group and si-CDH17+Nos treatment group (all P<0.01).The effect of si-CDH17+Nos treatment was more significant than that of si-CDH17 (P<0.01). Conclusion: Nos induces apoptosis and inhibits the migration of human colon cancer SW480 cells, which may be related to the down-regulation of CDH17 expression and inhibition of the Wnt3a/β-catenin signaling pathway.