1.Xinfukang drug-containing serum on bone marrow stem cell transcription and secretion of SDF-1α
Bohong KAN ; Yijing WANG ; Xiantong HU ; Qing GAO ; Yingchang FAN ; Yingna HE ; Kongyin ZHAO
International Journal of Biomedical Engineering 2014;37(5):275-278
Objective To clarify the effects of Xinfukang containing-serum on stromal cell-derived factor-1α (SDF-1α) translation and protein secretion of bone marrow stem cells (BMSCs).Methods BMSCs were isolated and amplified using bone marrow culture method,and were identified by flow cytometry.mRNA and protein secretion of SDF-1α were detected by quantitative PCR (q-PCR) and enzyme linked immunosorbent assay (ELISA),respectively.Results The expression of SDF-1α mRNA were significantly increased after 72 h in drug-containing serum,and SDF-1α mRNA in the experimental group was approximately 200 times as that in the control group (P<0.05).Secretion of SDF-1 α in the experimental group (277.561 1 ± 15.651 8) pg/ml was nearly doubled compared with that in the control group (153.107 1±14.765 1) pg/ml (P<0.05).Conclusions BMSCs from whole bone marrow adherent culture have high purity,and drug-containing serum can promote BMSCs to express SDF-1 α mRNA and secretion of SDF-1 α.
2.Study on the Experience of SUN Jie with"Theory of Sweat Pore"in Treating Dysuria Based on the Grounded Theory
Yao-Kan WU ; Bohong CAI ; Jie SUN
Journal of Zhejiang Chinese Medical University 2024;48(7):807-812
[Objective]To understand"theory of sweat pore(Xuanfu)"and its relationship with dysuria,as well as explore Professor SUN Jie's clinical experience in using"theory of sweat pore"to treat dysuria.[Methods]Through the research method of grounded theory,this paper analyzed and constructed a theoretical model that used"theory of sweat pore"to differentiate and treat the dysuria.The clinical documents of 135 cases of"dysuria"came from Professor SUN Jie's clinical diagnosis and treatment database.Besides,according to the qualitative research method by grounded theory,it concluded the core and built the theoretical framework by the three-level coding analysis.[Results]This study concluded that Professor SUN Jie summarized his experience to conclude the core category of"paying attention to the deficiency and excess of sweat pore,regulating and promoting Qi and body fluid".Two categories of"treatment based on syndrome differentiation with theory of sweat pore"and"treatment based on the principle"of dredging and assisting sweat pore were extracted.Based on this,Professor SUN Jie's experience in distinguishing and treating dysuria was derived,which focused on syndrome differentiation and treatment with sweat pore and on distinguishing the deficiency and stagnation of sweat pore and the nature of evil stagnation,and establishing treatment principle of dredging and assisting sweat pore,dispelling evil and supplementing deficiency as the treatment method.When using herbs,he used the prescriptions represented by Cassia Twig and Poria Cocos as the core,and combined alleviating water to dredge and assist sweat pore,warming and dredging to assist sweat pore,particularly good at using herbs of activating Yang.[Conclusion]Based on the research and summary of"theory of sweat pore",this paper summarized Professor SUN Jie's experience in using"theory of sweat pore"to treat dysuria,and constructed a theoretical model for Professor SUN Jie's use of"theory of sweat pore"in the treatment of dysuria.
3.The effect of miR⁃9 on the proliferation of neural stem cells by regulating β⁃tubulin Ⅲ and GFAP
Yu Wang ; Lan Zhao ; Bohong Kan ; Huiyan Shi
Acta Universitatis Medicinalis Anhui 2022;57(1):10-14
Objective :
To explore whether miR⁃9 affects the proliferation of neural stem cells (NSCs) by regulating β ⁃tubulin Ⅲ and glial fibrillary acidic protein (GFAP) .
Methods :
NSCs cells were isolated and cultured on the purchased healthy pregnant mice and the isolated cells were divided into NO group ( non⁃transfected NSCs cell line) , NN group (NSCs transfected with miR⁃9 ⁃NC) , NM group (NSCs transfected) transfected with miR⁃9 mimics) , NI group ( transfected with miR⁃9 inhibitor) . Nestin was identified by immunofluorescence , the contents of Mir⁃9 , β ⁃tubulin Ⅲ and GFAP were measured by QRT⁃PCR , the proliferation of NSCs was measured by MTT method , the apoptosis of NSCs was measured by flow cytometry , and the protein expressions of Mir⁃9 , β ⁃tubulin Ⅲ and GFAP were detected by Western blot.
Results :
The detection index Nestin of NSCs for 24 h after cell passage was positive in 90% of the NSCs , and co⁃localized with the nucleus , indicating the successful isolation of NSCs. The expression of miR⁃9 mRNA in NSCs of the NM group was the highest among the four groups , indicating that the transfection was successful. Among the four groups , β ⁃tubulin Ⅲ mRNA and protein expression was the highest in the NM group , and the GFAP mRNA and protein expression was the lowest. The NI group was the opposite (P < 0. 05) . In the four groups after 24 h , the cell activity of NSCs in the NM group was the strongest and showed a trend of gradually increasing with time (P < 0. 05) . The cell activity of NI was significantly weaker than that of the other three groups (P < 0. 05) . Among the four groups , the apoptosis rate of the NM group was lower than that of the other three groups (P < 0. 05) , and the apoptosis rate of NI was higher than that of the other three groups (P < 0. 05) . The difference in the above indicators between the NO group and the NN group was smaller.
Conclusion
Overexpressed miR⁃9 promotes the proliferation of NSCs by promoting the expression of β ⁃tubulin Ⅲ and inhibiting the expression of GFAP.