1.Quality control of recombinant oncolytic adenovirus/p53.
Kai GAO ; Hua BI ; Youxue DING ; Yonghong LI ; Chunmei HAN ; Ying GUO ; Chunming RAO
Acta Pharmaceutica Sinica 2011;46(12):1476-82
To establish a detection method of oncolytic adenovirus/p53 and standard of quality control, human telomerase reverse transcriptase (hTERT) promoter, CMV fusion promoter containing hypoxia reaction element (HRE) and p53 gene were identified by vector DNA restriction enzyme digestion and PCR analysis. The result conformed that all modified regions were in consistent with theoretical ones. Particle number was 2.0 x 10(11) mL(-1) determined by UV (A260). Infectious titer was 5.0 x 10(10) IU mL(-1) analyzed by TCID50. In vitro p53 gene expression in human lung cancer cell H1299 was determined by ELISA, and A450 ratio of nucleoprotein in virus infection group to control group was 5.2. Antitumor potency was evaluated by cytotoxicity assay using human lung cancer cell A549, and the MOI(IC50) of this gene therapy preparation was 1.0. The tumor cells targeted replication ability of recombinant virus was determined by TCID50 titer ratio of filial generation virus between human lung cancer cell A549 and human diploid epidermal fibrolast BJ cells after infected by virus with same MOI. TCID50 titer ratio of tumor cell infection group to normal cell infection control group was 398. The IE-HPLC purity of virus was 99.5%. There was less than 1 copy of wild type adenovirus within 1 x 10(7) VP recombinant virus. Other quality control items were complied with corresponding requirements in the guidance for human somatic cell therapy and gene therapy and Chinese pharmacopeia volume III. The detection method of oncolytic adenovirus/p53 was successfully established for quality control standard. The study also provided reference for quality control of other oncolytic viral vector products.
2.Practice and Exploration on Building a Financial Supervision System to Promote High-Quality Development of Public Hospitals
Yujiao YANG ; Chunmei BI ; Tao XU
Chinese Health Economics 2023;42(12):104-107
Public hospitals are the mainstay of China's healthcare service system and play a crucial role in promoting the construction of a Healthy China.A scientific financial and accounting supervision system can effectively ensure the realization of the financial and accounting supervision objectives of public hospitals.Through the construction of the"1455"framework,it determines the goals and requirements for financial supervision,clarifies the functions,scope,and path of financial supervision.Taking China-Japan Friendship Hospital as an example,it summarizes the innovative practices and achievements in improving systems,strengthening budget management,and promoting digitalization,based on the five major systems and five functions of the supporting framework.It provides practical experience for the establishment and improvement of the financial supervision system in public hospitals,promotes the realization of the goals of financial supervision in the healthcare industry,and advances the modernization of the healthcare governance system and governance capabilities.
3.Sulforaphane protects human platelets from high glucose-induced cellular apoptosis through down-regulating PI3K/Akt signaling pathway
Chunmei ZHANG ; Xinhui HUANG ; Jinqiu HU ; Xiaoyan BI ; Fuli YA
The Journal of Practical Medicine 2024;40(18):2530-2536
Objective To investigate the protective effects of Sulforaphane(SFN)against high glucose(HG)-induced human platelet apoptosis and elucidate the underlying mechanisms in vitro.Methods Purified platelets obtained from healthy individuals were pre-incubated with various concentrations of SFN(5,10,or 20 μmol/L)or a vehicle control(0.05%DMSO)for 40 minutes at 37℃.Subsequently,the platelets were stimulated with normal glucose(NG,5 mmol/L)or high glucose(HG,25 mmol/L)for an additional duration of 90 minutes.Flow cytometry was employed to assess platelet mitochondrial membrane potential depolarization(Δψm),exposure of phosphatidylserine(PS),and generation of total intraplatelet reactive oxygen species(ROS).Phosphorylation levels of PI3K and Akt were determined using Western blot.Results Compared to NG-treated human platelets,HG significantly induced depolarization of platelet Δψm and exposure of PS(P<0.001).These effects of HG were markedly attenuated by various concentrations of SFN(P<0.001).Mechanistically,SFN down-regulated the phosphorylation levels of PI3K(P<0.01)and Akt(P<0.05),which were increased by HG when compared to the vehicle control,and substantially reduced total intracellular ROS levels(P<0.001).The inhibitory effects of SFN on HG-induced phosphorylation of PI3K and Akt,as well as its efficacy on total intracellular ROS generation,Δψm depolarization,and PS exposure in HG-stimulated human platelets were completely reversed by a specific agonist for PI3K,740 Y-P.Conclusions The current study demonstrates that SFN exerts a protective effect on platelet apoptosis induced by HG,potentially through the down-regulation of the PI3K/Akt-mediated pathway in human platelets in vitro.These findings suggest that SFN may hold promise for improving thrombosis in diabetes mellitus and related chronic metabolic diseases.