1.Construction and characterization of liposomal magnetofection system in pig kidney cells.
Wenjie CHEN ; Haixin CUI ; Xiang ZHAO ; Jinhui CUI ; Yan WANG ; Changjiao SUN
Chinese Journal of Biotechnology 2014;30(6):972-981
Magnetic nano gene vector is one of the non-viral gene vectors, modified by functional group to bind cationic transfect reagents. Coupling magnetofection with the universal lipofection we developed a novel somatic cell transfection method as the so-called liposomal magnetofection (LMF). This approach is potential to provide somatic cell cloning with stable genetic cell lines to cultivate transgenic animals. In order to construct such liposomal magnetic gene vectors complexes system, we used nano magnetic gene vector to combine with liposomal cationic transfect reagents by molecular self-assembly. This vectors system successfully carried exogenous gene and then transfected animal somatic cells. Here, we conducted atomic force microscopy (AFM), zeta potential-diameter analysis and other characterization experiments to investegate the size distribution and morphology of magnetic nanoparticles, the way of the vectors to load and concentrate DNA molecules. Our data reveal that, the LMF of Pig Kidney cells exhibited higher transfection efficiency comparing with the transfection mediated by the commercial lipofectamine2000. Moreover, LMF method overcomes the constraint of transient expression mediated by lipofection. Meanwhile, MTT assay showed low cytotoxicity of LMF. Hence, LMF is a feasible, low cytotoxic and effective method of cell transfection.
Animals
;
Cations
;
Cell Line
;
DNA
;
Genetic Vectors
;
Kidney
;
cytology
;
Liposomes
;
Magnetics
;
Nanoparticles
;
Swine
;
Transfection
3.Establishment and assessment of the early maternal separation combined adult chronic unpredictable mild stress depression animal model
Chongkun SHEN ; Rong ZHANG ; Kerun CAO ; Haixin MO ; Ran YE ; Yongfei CUI ; Zhaoyang DONG ; Yafei SHI
Chinese Journal of Behavioral Medicine and Brain Science 2020;29(5):464-470
Objective:To explore the establishment and assessment of the depression model of adult SD rats induced by early maternal infant separation stress combined with adult chronic unpredictable mild stress(CUMS), and to observe the behavior and synaptic protein expression of SD rats.Methods:Twenty-four male rats were randomly divided into three groups: control group (CON group), chronic unpredictable stress group (CUMS group), maternal infant separation combined with chronic unpredictable stress group (MS + CUMS group). The depression model of rats in cums group was established by CUMS, and the depression model of rats in MS + CUMS group was established by maternal infant separation + CUMS. The depression model was evaluated by body mass and appearance observation, sugar water preference experiment, open field test and forced swimming test.Synapse-associated proteins (PSD-95 and synaptophysin (Syn)) of the hippocampus were assayed using Western blot. Syn protein expression was measured by immunohistochemistry.Results:After modeling, compared with CON group, the rats in CUMS group and MS + CUMS group had significantly lower body mass((126.43±3.88) g, (91.04±3.85) g, (69.89±6.67) g; t=5.03, 8.03, both P<0.01), significantly lower sugar water preference((94.21±0.56)%, (79.30±1.13)%, (72.73±1.82)%; t=8.24, 11.87, both P<0.01) and significantly increased immobility time((3.50±0.84)s, (13.59±2.40)s, (15.70±2.97)s; t=3.16, 3.82, both P<0.01). Among them, the body mass and sugar water preference of MS + CUMS group were lower than those of CUMS group ( t=3.00, 3.63; both P<0.01). (2)The center time of the rats in MS+ CUMS group markedly reduced compared with that of CON group((21.41±4.65) s, (8.96±2.37) s; t=2.66, P<0.05). (3)In the immunohistochemistry experiment, compared with the CON group, the percentage of Syn positive area in hippocampus of rats in CUMS group and MS+ CUMS group decreased significantly ((24.42±0.76)%, (14.00±0.95)%, (10.38±1.38)%; t=6.93, 9.33, P<0.01). (4)The results of Western blot showed that compared with CON group, both the expression of PSD-95 ((1.18±0.02), (0.74±0.06), (0.52±0.05), t=6.29, 9.31; both P<0.05) and Syn ((1.12±0.08), (0.95±0.06), (0.90±0.07); t=3.10, 4.04; both P<0.05) in hippocampus of rats in CUMS group and MS+ CUMS group were significantly decreased.And compared with the CUMS group, the expression of PSD-95 in MS+ CUMS group was lower( t=0.93, P<0.05). Conclusion:The method of MS combined with CUMS can effectively induce depression like behavior and decrease the expression of synaptic plasticity related protein of hippocampus in rats.
4.Correlations among negative emotion, coping style and social support in patients with total laryngectomy
Kezhen XIAO ; Fei LI ; Shaojuan CUI ; Haixin LONG ; Rui LIU ; Fang NAN
Chinese Journal of Modern Nursing 2019;25(12):1554-1557
Objective? To explore the mental state of laryngeal carcinoma patients after total laryngectomy and to analyze its correlations with coping style and social support. Methods? From January 2017 to October 2017, we selected 36 laryngeal carcinoma patients with total laryngectomy in otolaryngology dead and neck surgery at Beijing Tongren Hospital, Capital Medical University as subjects by convenience sampling. All of the patients were investigated with the Self-rating Depression Scale (SDS), Self-rating Anxiety Scale (SAS), Medical Coping Modes Questionnaire (MCMQ) and Social Support Rating Scale (SSRS). A total of 36 questionnaires were sent out and 34 of them were collected. Results? Among 34 patients, the total scores of SAS and SDS were (45.38±10.27) and (50.32±11.82) respectively. The score of "avoidance" coping style of patients after total laryngectomy was higher than that of norm with a statistical difference (P< 0.05). Pearson correlation analysis showed that SAS and SDS were negatively correlated with the total score of social support (P<0.05);SAS and SDS were positively correlated with the confrontation dimension of coping style (P<0.05), and were negatively correlated with the avoidance dimension (P< 0.05). Conclusions? After total laryngectomy, patients are with high levela of anxiety and depression. Medical staff should guide patients to take correct coping styles after surgery to face diseases, and improve patients' social support to relieve patients' negative emotions and total laryngectomy,and to promote the rehabilitation of diseases.