Objective To investigate effects of diabetes mellitus (DM) on pharmacokinetics (PK) of cephradine (CED).Methods DM model was induced by iv.alloxan 60 mg·kg-1.A reversed phase HPLC internal standard method was developed for measurement of CED plasma concentration.After blood collection,rats were sacrificed to collect kidneys for calculating kidney index(KW/BW).DM and normal control (CTL) rats were randomly assigned to receive iv.or ig.CED at a dose of 180 or 90 mg·kg-1.The 3p97 program was used to calculate PK parameters.Results The developed HPLC method was validated to have high specificity,precision,recovery and good storage stability,and met requirements for PK study of CED.The CED in rats of both DM and CTL groups showed the iv.two-compartment PK and ig.one-compartment PK and followed the first-order kinetics.Following iv.dosing,a remarkably decreased t1/2β and MRT,increased CLt were evident in DM group as compared with CTL group (P ＜ 0.05).After ig dosing,a significant decrease in t1/2k and t a remarkable increase in CLt and Cm=were observed for DM group as compared with CTL group (P ＜ 0.05).The DM rats showed a trend of decreased t1/2ka vs CTL rats.There was no significant difference in the oral bioavailability between the two groups (P ＞ 0.05).KW and KW/BW in DM group were increased remarkably compared with CTL group (P ＜ 0.05).Conclusion The DM vs CTL results in faster absorption and elimination of CED in rats,but does not have significantly affect in oral bioavailability.The compensatory hypertrophy and hyperfunction of early-stage diabetic kidneys may constitute one of causes of quick elimination of CED in rats with DM.

To investigate the cytotoxicity of the homemade peptide cationic liposome CDO14 and its efficacy of RNA interference (RNAi). MTT method was used to determine the cytotoxicity of the liposome to a human lung cancer cell line Luc-A549 that can express luciferase stably. Luciferase siRNA (Luc-siRNA) was transfected into Luc-A549 cells by CDO14. Contents of luciferase in the transfected cells were detected by luminous instrument and contents of total protein in these cells were detected by BCA method. Nude mice were inoculated with Luc-A549 cells in axilla to establish xenograft tumor model. Complexes of Luc-siRNA and the cationic liposomes were injected into the modeling mice via tail vein. Contents of luciferase in the transfected mice were detected by the whole body imaging system. The cytotoxicity of the homemade cationic liposome was similar to that of commercial liposome DOTAP, and lower than that of Lipo2000. The siRNA transfection efficacy mediated by CDO14 was higher than that mediated by DOTAP. The homemade peptide cationic liposome CDO14 is expected to serve as delivery vector in gene therapy because of its low cytotoxicity and high transfection efficiency.
Animals ; Cations ; Cell Line, Tumor ; Fatty Acids, Monounsaturated ; Genetic Therapy ; Genetic Vectors ; Humans ; Liposomes ; Luciferases ; Lung Neoplasms ; Mice ; Mice, Nude ; Peptides ; Quaternary Ammonium Compounds ; RNA Interference ; RNA, Small Interfering ; Transfection

Digital therapeutics(DTs)refers to a non-drug intervention method that uses electronic devices such as computers,smartphones,and wearable devices to evaluate and intervene through software programs and Internet technologies.It has been confirmed that there is a good therapeutic effect on a variety of mental disorders.Digital therapeutics can improve the insomnia problems of insomniacs,enhance the attention and work memory ability of patients with attention deficit hyperactivity disorder,and can also alleviate symptoms such as depression and anxiety disorder.Digital therapy will develop towards personalized treatment,popular treatment,fragmented treatment,and entertainment treatment in the future and have broad development prospects.

Objective: To analyze the gene characterization on the first imported D8 genotype measles virus in Liaoning province. Methods: In this study, Vero/Slam cells were used to isolate measles viruses from throat swabs. Fragments of the H gene (1854 nucleotides) and N gene (450 nucleotides) were amplified by reverse transcription-polymerase chain reaction (RT-PCR) and the PCR products were sequenced and analyzed. Results: The measles virus isolates and World Health Organization (WHO) D8 genotype reference strain (MVi/Manchester.GBR/30.94) belonged to the same branch in the genetic relationship tree. The nucleotide homology of the N and H gene was 98.9%. Phylogenetic trees were constructed with reference strains of the genotype D8 measles virus of China downloaded from GenBank. The result showed that the nucleotide similarities between the measles virus isolated in this study and the D8 genotype measles virus prevalent in Hong Kong from 2012 to 2016 and MVi/LosPatios.COL/11.18/D8 in Columbia was 100%. Conclusions It is the first time to do surveillance for the D8 genotype measles virus since measles virus surveillance was carried out. It was of great significance to accumulate the bases of measles virus molecular epidemiology in Liaoning province, and helpful to analyze and trace the transmission of measles virus in the whole country and the world.

Exonuclease Ⅷ (Exo Ⅷ), an ATP-independent dsDNA 5'-3' exonuclease, is a candidate protein with great application value for in vitro DNA recombination. However, the application of Exo Ⅷ in DNA recombination in vitro has not been reported. In this study, the recombinant expression vector of the truncated Exo Ⅷ (tExo Ⅷ) with the full exonuclease activity was built and used to achieve the overexpression of tExo Ⅷ in Escherichia coli. Based on the purified tExo Ⅷ protein with high-purity, the feasibility of tExo Ⅷ applied in vitro DNA recombination and effects of the reaction temperatures, reaction duration, and homology arm lengths were examined. The results showed that tExo Ⅷ was highly expressed in soluble form in E. coli. One liter of bacterial culture yielded 92.40 mg of purified tExo Ⅷ with the specific activity of 1.21×10⁵ U/mg. In a 10 μL recombination system containing 2.5 U tExo Ⅷ, the highest cloning efficiency was achieved in a reaction at 25 °C for 12.5 min and followed by incubation at 50 °C for 50 min. With addition of Pfu DNA polymerase, the homology arm extension strategy can effectively improve the recombination efficiency. Using competent E. coli Mach1 T1 with 2.2×10⁶ cfu/μg transformation efficiency as recipient cell, the recombination of a 1 kb fragment with a 21 bp homology arm and a 5.8 kb linearized vector can form about 1.1×10⁴ recombinant clones per μg vector, and the positive rates was over 80%. The recombination efficiency was increased with the increasing length of homology arm ranged from 8 to 21 bp. Under the optimal reaction condition, only 8 bp homology arm can still achieve valid DNA recombination. This novel in vitro DNA recombination system mediated by tExo Ⅷ was particularly characterized by its easy preparation, no limitation on restriction sites and high recombination cloning efficiency. All results revealed that the new efficient gene cloning system has potential application in the field of molecular biology.
Cloning, Molecular ; Escherichia coli ; genetics ; Exonucleases ; genetics ; Recombinant Proteins ; metabolism ; Recombination, Genetic

Objective: To analyze the genetic characterization of 2B genotype rubella virus strains first isolated in Liaoning province. Methods: Vero/Slam cells were used to isolate rubella viruses from throat swabs. Fragments of the E1 gene (739 nucleotides) were amplified by reverse transcription-polymerase chain reaction (RT-PCR) and the PCR products were sequenced and analyzed. The phylogenetic trees were constructed with rubella 2B genotypes reference strains. Results: RVi/Liaoning.CHN/13.18/1[2B] was isolateded from the case which had a history of international travel and probably imported from abroad. RVi/Liaoning.CHN/13.18/1[2B] was in the same 1ineage with RVs/Edinburgh.GBR/6.12[2B] and RVs/Kawasaki.JPN/17.11/1[2B], but far from the epidemic strains isolated in China in recent years. RVi/Liaoning.CHN/14.18/1[2B] belonged to the same lineage with rubella virus 2B genotype strains isolated in China from 2014 to 2015, and had the highest homology with RVi/Chouzhou.Anhui.CHN/17.15/2[2B] and RVi/Shaoyang.Zhejiang. CHN/13.13[2B]. Conclusions It was the first time to isolate the 2B genotype rubella virus in Liaoning province and the strain belonged to different 1ineage.

5-Hydroxytryptamine (5-HT) type 3 receptor (5-HT₃R) is the only type of ligand-gated ion channel in the 5-HT receptor family. Through the high permeability of Na⁺, K⁺, and Ca²⁺ and activation of subsequent voltage-gated calcium channels (VGCCs), 5-HT₃R induces a rapid increase of neuronal excitability or the release of neurotransmitters from axon terminals in the central nervous system (CNS). 5-HT₃Rs are widely expressed in the medial prefrontal cortex (mPFC), amygdala (AMYG), hippocampus (HIP), periaqueductal gray (PAG), and other brain regions closely associated with anxiety reactions. They have a bidirectional regulatory effect on anxiety reactions by acting on different types of cells in different brain regions. 5-HT₃Rs mediate the activation of the cholecystokinin (CCK) system in the AMYG, and the γ‍-aminobutyric acid (GABA) "disinhibition" mechanism in the prelimbic area of the mPFC promotes anxiety by the activation of GABAergic intermediate inhibitory neurons (IINs). In contrast, a 5-HT₃R-induced GABA "disinhibition" mechanism in the infralimbic area of the mPFC and the ventral HIP produces anxiolytic effects. 5-HT₂R-mediated regulation of anxiety reactions are also activated by 5-HT₃R-activated 5-HT release in the HIP and PAG. This provides a theoretical basis for the treatment of anxiety disorders or the production of anxiolytic drugs by targeting 5-HT₃Rs. However, given the circuit specific modulation of 5-HT₃Rs on emotion, systemic use of 5-HT₃R agonism or antagonism alone seems unlikely to remedy anxiety, which deeply hinders the current clinical application of 5-HT₃R drugs. Therefore, the exploitation of circuit targeting methods or a combined drug strategy might be a useful developmental approach in the future.
Serotonin ; Receptors, Serotonin, 5-HT3 ; Anxiety ; Neurons ; gamma-Aminobutyric Acid

Gypenosides, structurally analogous to ginsenosides and derived from a sustainable source, are recognized as the principal active compounds found in Gynostemma pentaphyllum, a Chinese medicinal plant used in the treatment of the metabolic syndrome. By bioactive tracking isolation of the plants collected from different regions across China, we obtained four new gypenosides (1-4), together with nine known gypenosides (5-13), from the methanol extract of the plant. The structures of new gypenosides were elucidated by one-dimensional (1D) and two-dimensional (2D) nuclear magnetic resonance (NMR) spectra, complemented by chemical degradation experiments. Through comprehensive evaluation involving COL1A1 promoter assays and PP2Cα activity assays, we established a definitive structure-activity relationship for these dammarane-type triterpenoids, affirming the indispensability of the C-3 saccharide chain and C-17 lactone ring in effectively impeding extracellular matrix (ECM) deposition within hepatic stellate cells. Further in vivo study on the CCl₄-induced liver damage mouse model corroborated that compound 5 significantly ameliorated the process of hepatic fibrosis by oral administration. These results underscore the potential of dammarane-type triterpenoids as prospective anti-fibrotic leads and highlight their prevalence as key molecular frameworks in the therapeutic intervention of chronic hepatic disorders.
Animals ; Mice ; Gynostemma ; Liver Cirrhosis/drug therapy* ; Triterpenes/pharmacology* ; Ginsenosides ; Extracellular Matrix ; Dammaranes

One major strategy to generate genetically modified mouse models is gene targeting in mouse embryonic stem (ES) cells, which is used to produce gene-targeted mice for wide applications in biomedicine. However, a major bottleneck in this approach is that the robustness of germline transmission of gene-targeted ES cells can be significantly reduced by their genetic and epigenetic instability after long-term culturing, which impairs the efficiency and robustness of mouse model generation. Recently, we have established a new type of pluripotent cells termed extended pluripotent stem (EPS) cells, which have superior developmental potency and robust germline competence compared to conventional mouse ES cells. In this study, we demonstrate that mouse EPS cells well maintain developmental potency and genetic stability after long-term passage. Based on gene targeting in mouse EPS cells, we established a new approach to directly and rapidly generate gene-targeted mouse models through tetraploid complementation, which could be accomplished in approximately 2 months. Importantly, using this approach, we successfully constructed mouse models in which the human interleukin 3 (IL3) or interleukin 6 (IL6) gene was knocked into its corresponding locus in the mouse genome. Our study demonstrates the feasibility of using mouse EPS cells to rapidly generate mouse models by gene targeting, which have great application potential in biomedical research.