2.Preparation and in vitro release characteristics of nerve growth factor thermosensitive gel for treating deafness
Zilan ZHU ; Wei LIU ; Laiyou WANG ; Zhiqing PANG ; Xuesong YU
Chinese Journal of Primary Medicine and Pharmacy 2014;21(8):1121-1123,后插2
Objective To prepare polypeptide nerve growth factor (NGF) thermosentitive gel and observe drug release in vitro,in order to provide scientific information for biopharmaceuticals delivery system design aiming to treat the inner disease.Methods The thermosensitive in situ gel was prepared with NGF as main component and Pluronic F127 as the gel matrixes.The effect of concentration of gel matrix PF127 on lower critical solution temperature(LCST) were investigated.In vitro release kinetic studies were performed using membraneless dissolution method and reversed-phase high performance liquid chromatography (HPLC) method was adopted to determine NGF content in the dissolution medium.Results The average LCST of NGF-loaded gel prepared by different concentration of PF127 was 28.48-36.26℃ and the gels had good stability.The in vitro release kinetics was well characterized by sustained release and can be fitted by zero order kinetics.The in vitro accumulated release ratio of NGF in the thermosensitive gel reached to above 95%.hβ-NGF loaded in higher PF127 gel resulted in a more sustained release of hβ-NGF from the thermosensitive gel.Conclusion Well-prepared NGF thermosensitive gel is a promising inner ear-oriented drug delivery system for treating deafness and deserves to further development.
3.Draft Genome Assembly and Annotation for Cutaneotrichosporon dermatis NICC30027, an Oleaginous Yeast Capable of Simultaneous Glucose and Xylose Assimilation
Laiyou WANG ; Shuxian GUO ; Bo ZENG ; Shanshan WANG ; Yan CHEN ; Shuang CHENG ; Bingbing LIU ; Chunyan WANG ; Yu WANG ; Qingshan MENG
Mycobiology 2022;50(1):66-78
The identification of oleaginous yeast species capable of simultaneously utilizing xylose and glucose as substrates to generate value-added biological products is an area of key economic interest. We have previously demonstrated that the Cutaneotrichosporon dermatisNICC30027 yeast strain is capable of simultaneously assimilating both xylose and glucose, resulting in considerable lipid accumulation. However, as no high-quality genome sequencing data or associated annotations for this strain are available at present, it remains challenging to study the metabolic mechanisms underlying this phenotype. Herein, we report a 39,305,439 bp draft genome assembly for C. dermatis NICC30027 comprised of 37 scaffolds, with 60.15% GC content. Within this genome, we identified 524 tRNAs, 142 sRNAs, 53 miRNAs, 28 snRNAs, and eight rRNA clusters. Moreover, repeat sequences totaling 1,032,129 bp in length were identified (2.63% of the genome), as were 14,238 unigenes that were 1,789.35 bp in length on average (64.82% of the genome). The NCBI non-redundant protein sequences (NR) database was employed to successfully annotate 11,795 of these unigenes, while 3,621 and 11,902 were annotated with the Swiss-Prot and TrEMBL databases, respectively. Unigenes were additionally subjected to pathway enrichment analyses using the Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), Cluster of Orthologous Groups of proteins (COG), Clusters of orthologous groups for eukaryotic complete genomes (KOG), and Non-supervised Orthologous Groups (eggNOG) databases.Together, these results provide a foundation for future studies aimed at clarifying the mechanistic basis for the ability of C. dermatis NICC30027 to simultaneously utilize glucose and xylose to synthesize lipids.
4.Effects and Mechanism of Oncolytic Virus M 1 Inducing the Apoptosis of Cervical Cancer C- 33A Cells
Xiao XIAO ; Yasi ZHOU ; Chuyin PENG ; Jinqing DENG ; Laiyou WANG ; Wenbo ZHU
China Pharmacy 2021;32(23):2827-2831
OBJECTIVE:To study the effects and mechanism of oncolytic virus M 1(called M 1 virus for short )inducing the apoptosis of cervical cancer C-33A cells. METHODS :MTT assay was used to detect survival rate of C- 33A cells that were treated with different titers (0,0.001,0.01,0.1,1,10 PFU/cell)of M 1 virus. C- 33A cells were divided into control group (0 PFU/cell), low-dose,medium-dose and high-dose groups of M 1 virus(0.001,0.01,0.1 PFU/cell). After treated with corresponding titers of M1 virus for 48 h,flow cytometry was used to detect the apoptotic rate and infection rate of cells;Western blot was performed to detect the protein expression of C/EBP homologous proteins (CHOP),caspase-12,caspase-3 and cleaved-caspase- 3. RESULTS : After treated with different titers of M 1 virus,the survival rate of C- 33A cells decreased significantly (P<0.01),and showed a dose-dependent tr end. Compared with control group ,the apoptotic rate and infection rate of cells in M 1 virus groups as well as the protein expression of CHOP ,caspase-12 and cleaved-caspase- 3(except for medium-dose group )in M 1 virus medium-dose and high-dose groups were increased significantly (P<0.01). CONCLUSIONS :M1 virus can induce the apoptosis of cervical cancer C-33A cells ,and its mechanism may be related to the activation of endoplasmic reticulum stress pathway.