Peptide-RNA complexation-induced fluorescence"turn on"displacement assay for the recognition of small ligands targeting HIV-1 RNA / 药物分析学报

The regulator of expression of virion(Rev)protein binds specifically to the Rev-responsive element(RRE)RNA in order to regulate the expression of the human immunodeficiency virus(HIV)-1 genes.Fluores-cence indicator displacement assays have been used to identify ligands that can inhibit the Rev-RRE interaction;however,the small fluorescence indicators cannot fully replace the Rev peptide or protein.As a result,a single rhodamine B labeled Rev(RB-Rev)model peptide was utilized in this study to develop a direct and efficient Rev-RRE inhibitor screening model.Due to photon-induced electron transfer quenching of the tryptophan residue on the RB fluorophore,the fluorescence of RB in Rev was weakened and could be dramatically reactivated by interaction with RRE RNA in ammonium acetate buffer(approximately six times).The interaction could reduce the electron transfer between tryptophan and RB,and RRE could also increase RB fluorescence.The inhibitor screening model was evaluated using three known positive Rev-RRE inhibitors,namely,proflavin,6-chloro-9-[3-(2-chloroethylamino)pro-pylamino]-2-methoxyacridine(ICR 191),and neomycin,as well as a negative drug,arginine.With the addition of the positive drugs,the fluorescence of the Rev-RRE decreased,indicating the displacement of RB-Rev.This was confirmed using atomic force microscopy(AFM)and the fluorescence was essentially unaffected by the addition of arginine.The results demonstrated that RB-Rev can be used as a fluorescent probe for recognizing small ligands that target RRE RNA.The Rev-RRE inhibitor screening model offers a novel approach to evaluating and identifying long-acting Rev inhibitors.

Clinical study on the 18F-FDG PET/CT " one-stop" evaluation of the volume and activity of epicardial adipose tissue / 中华核医学与分子影像杂志

Objective:To investigate the accuracy of free-breathing CT in evaluating the volume and shape of epicardial adipose tissue (EAT), and further explore the characteristics of EAT volume and activity in patients with atrial fibrillation using 18F-FDG PET/CT " one-stop" imaging. Methods:(1) Retrospective analysis was performed on 20 patients (16 males, 4 females, age: 33-86 (61.1±14.2) years) who underwent 18F-FDG PET/CT imaging and without obvious diseases affecting the images of the heart and surrounding lungs between March 2020 and May 2020 in Beijing Chaoyang Hospital. Free-breathing CT and breath-hold high resolution CT (HRCT) images were reviewed. Spearman rank correlation analysis, Bland-Altman consistency analysis and intraclass correlation coefficient (ICC) were used to evaluate the correlation and consistency of the EAT volume and shape, as well as the repeatability of the two operators′ measurements. (2) Prospective analysis was conducted to compare the differences in EAT volume and 18F-FDG uptake values between 20 patients (6 males, 14 females, age: 52-76 (66.0±6.4) years) with atrial fibrillation and 10 healthy controls (3 males, 7 females, age: 59-69 (66.0±3.6) years) collected between August 2017 and August 2018 in Beijing Chaoyang Hospital. Mann-Whitney U test was used to compare the differences in EAT volume and 18F-FDG SUV max between patients with atrial fibrillation and healthy controls. EAT volume measurement was conducted by the combination of Mimics Research 21.0 software and manual analysis. The shape of EAT was automatically calculated by the same software to obtain the maximum length of the projection of the three-dimensional (3D) model on the reference axes ( x, y, z). SUV max of EAT was manually measured. Results:The measurements of EAT volume had good repeatability (intra-operator ICC=0.999; inter-operator ICC=0.997). There was a good correlation and a good consistency between EAT volumes measured by free-breathing CT and breath-hold HRCT (96.6 (79.9, 136.4) vs 96.2 (80.9, 135.8) ml; rs=0.929, P<0.001); data of 19 cases were within 95% limits of agreement (95% LoA). The maximum projection length of EAT 3D model on the reference coordinate axis also showed good correlation and consistency ( x axis: rs=0.869, P<0.001, data of 19 cases were within 95% LoA; y axis: rs=0.854, P<0.001, data of 18 cases were within 95% LoA; z axis: rs=0.586, P=0.007, data of 20 cases were within 95% LoA). EAT volume of atrial fibrillation group was higher than those of healthy control group (137.2 (113.9, 202.9) vs 94.4 (76.6, 134.4) ml; z=-2.11, P=0.035) and SUV max of EAT in the atrial fibrillation group was higher than that in healthy control group (1.2 (1.1, 1.5) vs 1.1 (1.0, 1.2); z=-2.14, P=0.035). Conclusions:Free-breathing CT and breath-hold HRCT have good correlation, consistency and repeatability in measurement of EAT volume and shape. 18F-FDG PET/CT can be a " one-stop" imaging strategy for the evaluation of EAT volume and activity.

The single nucleotide polymorphism rs1814521 in long non-coding RNA ADGRG3 associates with the susceptibility to silicosis: a multi-stage study

BACKGROUND@#This study aimed to evaluate the correlation between long non-coding RNA (lncRNA)-related single nucleotide polymorphisms (SNPs) and susceptibility to silicosis.@*METHODS@#First, RNA-sequencing (RNA-seq) data were comprehensively analyzed in the peripheral blood lymphocytes of eight participants (four silicosis cases and four healthy controls) exposed to silica dust to identify differentially expressed lncRNAs (DE-lncRNAs). The functional SNPs in the identified DE-lncRNAs were then identified using several databases. Finally, the association between functional SNPs and susceptibility to silicosis was evaluated by a two-stage case-control study. The SNPs of 155 silicosis cases and 141 healthy silica-exposed controls were screened by genome-wide association study (GWAS), and the candidate SNPs of 194 silicosis cases and 235 healthy silica-exposed controls were validated by genotyping using the improved Mutiligase Detection Reaction (iMLDR) system.@*RESULTS@#A total of 76 DE-lncRNAs were identified by RNA-seq data analysis (cut-offs: fold change > 2 or fold change < 0.5, P < 0.05), while 127 functional SNPs among those 76 DE-lncRNAs were identified through multiple public databases. Furthermore, five SNPs were found to be significantly correlated with the risk of silicosis by GWAS screening (P < 0.05), while the results of GWAS and iMLDR validation indicated that the variant A allele of rs1814521 was associated with a reduced risk of silicosis (OR = 0.76, 95% CI = 0.62-0.94, P = 0.011).@*CONCLUSION@#The presence of the SNP rs1814521 in the lncRNA ADGRG3 is associated with susceptibility to silicosis. Moreover, ADGRG3 was found to be lowly expressed in silicosis cases. The underlying biological mechanisms by which lncRNA ADGRG3 and rs1814521 regulate the development of silicosis need further study.

Gut microbiota and gestational diabetes mellitus: a review / 中华围产医学杂志

The pathogenesis of gestational diabetes mellitus (GDM) remains unclear. Recent studies have found that a dysbiosis of gut flora could lead to metabolism and immune system disorders in human beings, resulting in GDM. The gut microbiome changes can be affected by various factors, of which diet and antibiotic exposure are the most dominant. From the perspective of the gut microbiome changes, individualized dietary guidance, evidence-based administration of antibiotics, appropriate intake of prebiotics, and probiotics supplement may help prevent and treat GDM.

Research progress on the role of surfactant protein A and D in pulmonary fibrosis / 中国职业医学

Pulmonary fibrosis is an interstitial lung disease caused by different pathogenic factors. It has the characteristics of high morbidity and poor prognosis, which seriously affects the quality of life of patients. However, its pathogenesis has not yet been fully elucidated. Surfactant protein(SP)-A and SP-D are lipoprotein complexes distributed at the air-liquid interface of alveoli, synthesized and secreted by alveolar type Ⅱ epithelial cells and bronchiolar cells. They are important parts of the innate immune system, which participate in the host defense process through a variety of regulatory methods, and play an important role in regulating cell apoptosis and lung inflammation, promoting the process of epithelial repair and maintaining the stability of alveolar structure. The disorder and mutation of SP-A and SP-D may be the influencing factors of the pathogenesis of pulmonary fibrosis. Serum SP-A and SP-D levels are differentially expressed in patients with pulmonary fibrosis and normal healthy individuals, and are related to the severity of pulmonary fibrosis. They are considered to be a class of biomarkers that sensitively reflect lung epithelial cell damage and can be used in the diagnosis, treatment and prognostic evaluation of pulmonary fibrosis.

Construction of yeast cell factories for production of azadirachtin precursor tirucalla-7,24-dien-3β-ol / 中国中药杂志

Azadirachtin, as a botanical insecticide, is a highly oxidized limonoid triterpenoid existing in the seeds of Azadirachta indica. However, due to the low content in the seeds, the production of azadirachtin by seed extraction has low yield. Chemical synthesis of azadirachtin is characterized by complex process and low yield. Synthetic biology provides an alternative for the supply of azadirach-tin. In this study, two oxidosqualene cyclases AiOSC1 and MaOSC1 respectively derived from A. indica and Melia azedarach were identified in yeast. A yeast strain producing tirucalla-7,24-dien-3β-ol was constructed by integration of AiOSC1, Arabidopsis thaliana-derived squalene synthase gene(AtAQS2), and Saccharomyces cerevisiae-derived truncated 3-hydroxy-3-methyl-glutaryl coenzyme A reductase gene(PgtHMGR) into the delta site of yeast. Then, the function of MaCYP71BQ5 was successfully verified in yeast after this gene was introduced into the constructed yeast strain. This study not only laid a foundation for the biosynthesis of tirucalla-7,24-dien-3β-ol, but also provided a chassis cell for the functional identification of cytochrome oxidases(CYP450 s) in azadirachtin biosynthesis pathway.

Regulation of β-mercuryl alcohol metabolic flow in Saccharomyces cerevisiae cells / 中国中药杂志

In this study, citrate synthase gene(CIT2), and malate synthase gene(MLS1) were successfully knocked out in β-amyrin-producing yeast cells by using CRISPR/CAS9. The promoter of phosphoglucose isomerase gene(PGI1) was replaced by that of cytochrome c oxidase subunit Ⅶa(Cox9)to weaken its expression, aiming to channel more carbon flux into the NADPH-producing pathway. The fermentation results showed that CIT2 deletion had no effect on the β-amyrin production. Compared with the control strain, the production of β-amyrin was increased by 1.85 times after deleting MLS1, reaching into 3.3 mg·L~(-1). By replacing the promoter of PGI1, the β-amyrin yield was 3.75 times higher than that of the control strain, reaching up to 6.7 mg·L~(-1). This study successfully knocked out the CITT2 and MLS1 genes and weakened the PGI1 gene by using CRISPR/CAS9, which directly influenced the production of β-amyrin and provided some reference for the the metabolic engineering of triterpernoid producing strain.

Study of heterologous efficient synthesis of β-amyrin and high-density fermentation / 中国中药杂志

In this study, the synthetic pathway of β-amyrin was constructed in the pre-constructed Saccharomyces cerevisiae chassis strain Y0 by introducing β-amyrin synthase from Glycyrrhiza uralensis, resulting strain Y1-C20-6, which successfully produced β-amyrin up to 5.97 mg·L~(-1). Then, the mevalonate pyrophosphate decarboxylase gene(ERG19), mevalonate kinase gene(ERG12), 3-hydroxy-3-methylglutaryl-CoA synthase gene(ERG13), phosphomevalonate kinase gene(ERG8) and IPP isomerase gene(IDI1)were overexpressed to promoted the metabolic fluxto the direction of β-amyrin synthesis for further improving β-amyrin production, resulting the strain Y2-C2-4 which produced β-amyrin of 10.3 mg·L~(-1)under the shake flask fermentation condition. This is 100% higher than that of strain Y1-C20-6, illustrating the positive effect of the metabolic engineering strategy applied in this study. The titer of β-amyrin was further improved up to 157.4 mg·L~(-1) in the fed-batch fermentation, which was almost 26 fold of that produced by strain Y1-C20-6. This study not only laid the foundation for the biosynthesis of β-amyrin but also provided a favorable chassis strain for elucidation of cytochrome oxidases and glycosyltransferases of β-amyrin-based triterpenoids.

The relationship between serum and seminal plasma vitamin D and male reproductive function / 医学研究生学报

Objective Male infertility accounts for 40 to 50% of the total number of infertility in the world. Among many factors that cause male infertility, vitamin D is considered to be directly related to male fertility. The purpose of this study was to explore the relationship between serum and seminal plasma vitamin D and male reproductive function, and provide a more comprehensive research direction for studying the specific mechanism of vitamin D on male reproduction. Methods A total of 198 infertile males, receiving andrological examination from June 2017 to January 2018 in the Center for Reproductive Medicine, Jinling Hospital (Nanjing, China) was included in our study. Serum and seminal plasma vitamin D levels were measured by electrochemiluminescence immunoassay (ECLIA) kits. The associations between vitamin D and biomarkers of male reproduction were analyzed. Results Serum 25(OH) vitamin D level [26.17(19.61-31.99)ng/mL] was in positive relation with semen volume[3.8(3.1-4.8)ng/mL] (r=0.229,P=0.003). Seminal plasma 25(OH) vitamin D level was not related to serum 25(OH) vitamin D level, but in negative relation with sperm concentration(r=0.174,P=0.016) and positive relation with semen volume(r=0.271,P=0.0001). Serum 25(OH) vitamin D level was in positive relation with seminal plasma fructose concentration(r=0.256,P=0.002), total fructose content (r=0.310,P=0.0002) and total zinc content(r=0.26,P=0.002). The level of serum and seminal plasma vitamin D leve was not related to serum anti-Mullerian hormone(AMH), seminal plasma AMH, serum inhibin (INH B) and seminal plasma INH B(P>0.05). Conclusion Vitamin D is associated with affiliated gland function. The seminal vesicles and prostate produced by semen may be the main source of vitamin D in the male reproductive system.

Chemical screen identifies a geroprotective role of quercetin in premature aging

Aging increases the risk of various diseases. The main goal of aging research is to find therapies that attenuate aging and alleviate aging-related diseases. In this study, we screened a natural product library for geroprotective compounds using Werner syndrome (WS) human mesenchymal stem cells (hMSCs), a premature aging model that we recently established. Ten candidate compounds were identified and quercetin was investigated in detail due to its leading effects. Mechanistic studies revealed that quercetin alleviated senescence via the enhancement of cell proliferation and restoration of heterochromatin architecture in WS hMSCs. RNA-sequencing analysis revealed the transcriptional commonalities and differences in the geroprotective effects by quercetin and Vitamin C. Besides WS hMSCs, quercetin also attenuated cellular senescence in Hutchinson-Gilford progeria syndrome (HGPS) and physiological-aging hMSCs. Taken together, our study identifies quercetin as a geroprotective agent against accelerated and natural aging in hMSCs, providing a potential therapeutic intervention for treating age-associated disorders.

Investigation and optimization on ability of enzymatic hydrolysis of Mori Cortex residue / 中国中药杂志

Residue of Mori Cortex was studied to optimize its enzymatic hydrolysis process, and explore its potential as a carbon source for biochemistry and biofuel production. The cellulose content of diluted acid pretreated (DAP) and non-pretreated from Mori Cortex were measured in this study, and the results showed that the cellulose content of DAP and non-pretreated from Mori Cortex were 52.5% and 47%, respectively. This higher cellulose content indicated that residue of Mori Cortex had the potential to act as a carbon source for biochemistry and biofuel production. Enzymatic hydrolysis of pretreated and non-pretreated from Mori Cortex was conducted under different enzyme loading amount. 40 FPU·(g DW）⁻¹ enzyme loading was determined as the optimal amount by comparing the yield of sugar and the rate of enzymolysis. Under this condition, the concentrations of glucose, xylose, arabinose sugar were 23.82, 4.84, 3.6 g·L⁻¹, and the corresponding enzymatic hydrolysis rate was 45.33% which was 2.3 times higher than that of non-pretreated from Morus alba residues. Fed-batch enzymatic hydrolysis was conducted finally to get higher sugar yield, and the final glucose concentration reached up to 38 g·L⁻¹ with the enzymatic hydrolysis rate of 36.19%. The results indicated that Mori Cortex residue had higher cellulose and hemicellulose contents, so it had the potential to become a carbon source to produce the bio-chemicals and biofuels. Through enzymatic hydrolysis, it can be converted into microbial available monosaccharides; and through fermentation, it can be converted into high value-added chemicals, biofuels, etc., to solve the problem of residue pollution, and achieve the sustainable development and greening of Chinese pharmaceutical production process.

Influence of light on gene expression of key synthesis enzyme genes FtANR and FtLAR about proanthocyanidin in seeds of homologous plant of food and medicine Fagopyrum tataricum / 中国中药杂志

Tartary buckwheat Fagopyrum tataricum is an important medicinal and functional herb due to its rich content of flavonoids in the seeds. F.tataricum exhibited good functions for free radicals scavenging， anti-oxidation， anti-aging activities. Although much genetic knowledge of the synthesis， regulation， accumulation of rutin， the genetic basis of proanthocyanidins(PAs) in tartary buckwheat and their related gene expression changes under different lights(blue， red， far red， ultraviolet light) remain largely unexplored. In this study， we cloned one anthocyanidin reductase gene(ANR) and two leucocyanidin reductase gene(LAR) named FtANR，FtLAR1，FtLAR3 involved in formation of(+)-catechin and(-)-epicatechin precusor proanthocyanidin by digging out F. tataricum seed transcriptome data. The expression data showed that the opposite influence of red light on these gene transcript level compared to others lights. The expression levels of FtANR and FtLAR1 decreased and FtLAR3 appeared increment after exposed in the red light， while the expression levels of those genes appeared opposite result after exposed in the blue and far red light.

Chemical constituents of amides from Zanthoxylum planispinum / 中草药

Objective: To study the chemical constituents of Zanthoxylum planispinum. Methods: The compounds were isolated and purified by repeated silica gel column chromatography, Sephadex LH-20 gel column chromatography, and preparative HPLC. Their chemical structures were identified on the basis of physico-chemical properties and spectral analysis. Results: Seven amides were isolated and elucidated as (2E, 7E, 9E)-N-isobutyl-6,11-dioxo-2,7,9-dodecatrienamide (1), armatamide (2), pellitorine (3), lanyuamide I (4), tetrahydro-bungeanool (5), hydroxy-γ-isosanshool (6), and dihydrobungeanool (7). Conclusion: Compound 1 is a new unsaturated alkylamide named planispinamide, and compounds 2-7 are obtained from this plant for the first time.

Recent advances of synthetic biology for production of functional ingredients in Chinese materia medica / 中国中药杂志

The functional ingredients in Chinese materia medica are the main active substance for traditional Chinese medicine and most of them are secondary metabolites derivatives. Until now,the main method to obtain those functional ingredients is through direct extraction from the Chinese materia medica. However, the income is very low because of the high extraction costs and the decreased medicinal plants. Synthetic biology technology, as a new and microbial approach, can be able to carry out large-scale production of functional ingredients and greatly ease the shortage of traditional Chinese medicine ingredients. This review mainly focused on the recent advances in synthetic biology for the functional ingredients production.

Lengthen-stem bipolar-femur prosthetic replacement for the treatment of old patients with intertrochanteric fracture osteoporosis / 中国基层医药

Objective To evaluate the clinical application and significance of lengthen-stem bipolar-femur prosthetic replacement for the treatment of old-age patients with intertrochanteric fracture osteoporosis.Methods 28 cases of patients aging from 75 to 99 years old of intertrochanteric fracture osteoporosis treated with lengthen- stem cemented bipolar prosthesis were studicd from March 2000 to December 2006.After taking the blank margin, the bones of different sizes were replaced and the steel wire was fixed.After determining the depth of the front an- gle,the artificial bone was placed.Results After 28 examples attaining the following-up examination for 7 months to 3 years,with an average of 1.5 years,its function according to Harris standard was evaluated 3 months after the operation,8 examples were excellent,13 examples good,5 examples pass,2 examples inferior.The excellent or good rate reached 75% ,with no abnormal cases,no joint dislocation during the followed-up period.1 example had the phantom phenomenon 1 year after the operation.2 examples among the inferior had got more serious internal medicine disease which affected the restoring function.1 example died of the internal medicine disease 1 year after the operation.Conclusion By using the lengthen-stem bipolar-femur prosthetic replacement for the treatment of old patients with inrertrochanteric fracture osteoporosis,the patients will restore quickly after the operation and can carry a heavy load at an early time.The illness complication and the mortality rate will be redaced.But its related disease must be strictly dealt with and the surgery operating skill must be grasped.