Fragments of ? and ? genes were cloned into expression plasmid pET-21b. Bacteria harbouring expression plasmid was induced with IPTG,and a band of expressed protein of 16 kD was found in PAGE. They expressed as soluble products. ? expression product reached about 5% of total bacterial protein,? expression product reached about 15% of total bacterial protein. The products were confirmed by Western-blotting.

Objective:To review the chemical and pharmacological activities of Codonopsis lanceolata in order to provide reference for the further development of C. lanceolata. Methods:The related literatures at home and abroad in the past 40 years were reviewed and analyzed, and then the chemical components and pharmacological actions of C. lanceolata were summarized. Results: The major chemical constiturents in C. lanceolata were terpenoids, alkaloids, phenylethanoid glycoside and flavonoids. The pharmacological ac-tivities were antioxidation, anti-inflammatory, anti-tumor, antiplatelet aggregation, hypoglycemic and hypolipidemic effects, etc. Con-clusion:The review provides reference for the further development and comprehensive utilization of C. lanceolata. The development of relevant safe and effective agents is still needed, and at present, the definition of mechanism and the extension of clinical application remain as the primary tasks of the exploration of C. lanceolata.

In order to meet the requirements of medical teaching contents and curriculum system reform, we adopt the experience ofHarvard University School of Medicine to integrate Biochemistry and Cell Biology into a new one—— "Chemistry and Biology of theCell". We have carried out the integration of the two courses three times. In this new course, repeated contents have been reduced, thelinkage of the two courses has been increased, and the burden of students was lightened,which makes students contact the clinic earlierand enables to improve the students' ability to solve problems.

To investigate the function of testis sperm binding protein (TSBP) in sperm capacitation and acrosome reaction, the effect of the recombinant TSBP on the activity of protein kinase A was detected in the transfected cell line. With the use of prokaryotic expressing plasmid pGEX-5X-1/tsbp as template, the novel gene tsbp was amplified by PCR and a eukaryotic expressing vector pcDNA3.1/myc-His(-)B/tsbp was constructed. DNA sequencing and restriction endonuclease digestion analysis indicated that the eukaryotic expression vector pcDNA3.1/myc-His(-)B/tsbp had been constructed successfully. After the recombinant plamid being transfected into HEK293 cells, Western blot verified the expression of tsbp. Fusion protein His6-TSBP was purified from the cell lysate by immobilized metal-ion affinity chromatography (IMAC). Radioautograph revealed a higher PKA activity in the transfected HEK293 cells than in the control group, which indicates that TSBP can increase the activity of PKA.
Acrosome Reaction ; drug effects ; Cell Line ; Cyclic AMP-Dependent Protein Kinases ; drug effects ; Humans ; Male ; Recombinant Proteins ; biosynthesis ; genetics ; pharmacology ; Seminal Plasma Proteins ; biosynthesis ; genetics ; pharmacology ; Sperm Capacitation ; drug effects

Antibiotics are a category of chemical compounds used to treat bacterial infections and are widely applied in cultivation,animal husbandry,aquaculture,and pharmacy.Currently,residual antibiotics and their metabolites pose a potential risk of allergic reactions,bacterial resistance,and increased cancer incidence.Residual antibiotics and the resulting bacterial antibiotic resistance have been recognized as a global challenge that has attracted increasing attention.Therefore,monitoring antibiotics is a critical way to limit the ecological risks from antibiotic pollution.Accordingly,it is desirable to devise new analytical platforms to achieve efficient antibiotic detection with excellent sensitivity and specificity.Quantum dots(QDs)are regarded as an ideal material for use in the development of antibiotic detection biosensors.In this review,we characterize different types of QDs,such as silicon,chalcogenide,carbon,and other doped QDs,and summarize the trends in QD-based antibiotic detection.QD-based sensing applications are classified according to their recognition strategies,including molecularly imprinted polymers(MIPs),aptamers,and immunosensors.We discuss the advantages of QD-derived antibiotic sensors,including low cost,good sensitivity,excellent stability,and fast response,and illustrate the current challenges in this field.

Chromosomal integration of heterologous genes or pathways is preferred over the use of episomal plasmids for its inherently stability and thus more desirable in the industrial setting. However, the position of integration of heterologous genes in the genome influences the expression levels. In combination of high throughput transformation of the Yeast Knock-out Collection (YKO) and FACS analysis, the position effect on heterologous reporter gene gfp was identified across the whole genome in yeast. In total 428 high-expressed sites and 444 low-expressed sites were spotted, providing massive data to analyze patterns and reasons for region dependency of gene expression on the genome-wide scale.
Gene Expression Regulation, Fungal ; Gene Knock-In Techniques ; Genes, Reporter ; Genome, Fungal ; Saccharomyces cerevisiae ; genetics

The purpose of this study is to reveal the role of Girdin in regulating the aggregation of actin filaments by studying the relationship between PKB/Akt and Girdin. First we used Scansite software (http://scansite.mit.edu) to predict relevant target sites of PKB/Akt on mouse Girdin. To gain insight into the role of phosphorylation of Girdin by PKB/Akt, we assessed the location of phosphorylated Girdin in fertilized eggs by staining with anti-P-Girdin 1 417 Ab. We detected a distinct increase in the fluorescence signal of F-actin and P-Girdin 1 417 after microinjection of Akt WT and myr-Akt. The addition of myr-Akt induced phosphorylation of Girdin in mouse fertilized eggs. In addition, siRNA-mediated Akt-knockdown blocked phosphorylation of Girdin. The distribution of actin filaments was obviously scattered. These results strongly suggest that PKB/Akt could directly phosphorylate Girdin on Ser1 417 and promote its function in mouse fertilized eggs.
Actins ; physiology ; Animals ; Mice ; Microfilament Proteins ; physiology ; Phosphorylation ; Proto-Oncogene Proteins c-akt ; physiology ; RNA, Small Interfering ; Vesicular Transport Proteins ; physiology ; Zygote

ObjectiveTo explore the prevalence rate and related risk factors of anxiety， depression and insomnia symptoms in close contacts of COVID-19 patients undergoing centralized quarantine and observation， so as to provide references for follow-up targeted psychological crisis intervention. MethodsFrom February 6 to 8， 2020， a total of 136 close contacts at designated quarantine sites in four districts and counties of Ya'an city were enrolled， and assessed using the general information questionnaire， Hospital Anxiety and Depression Scale （HAD） and Insomnia Severity Index （ISI）. ResultsA total of 26 quarantined individuals （19.12%） had various degrees of anxiety， depression or insomnia， including 16 cases （11.76%） with anxiety， 9 cases （6.62%） with depression and 16 cases （11.76%） with insomnia. Correlation analysis showed that HAD （A） score was positively correlated with HAD （D） score （r=0.642， P<0.01）. HAD （A） score was positively correlated with ISI score （r=0.590， P<0.01）. HAD （D） score was positively correlated with ISI score （r=0.329， P<0.01）. Multivariate Logistic regression analysis showed that the risk factor of anxiety was the first week of isolation （OR=3.290， P<0.05）， meantime， the risk factors of insomnia included the old age （OR=7.990， P<0.05） and the first week of isolation （OR=4.659， P<0.05）. ConclusionThe close contacts of COVID-19 infections undergoing centralized quarantine and observation in Ya’an have high prevalence rate of anxiety and insomnia， and the symptoms are most severe during the first week of quarantine.