1.Effects of HO-1 modified rat bon e marrow mesenchymal stem cells on lymphocytes in vitro
Nannan FU ; Hongli SONG ; Benjuan WU ; Tao LIU
Chinese Journal of Microbiology and Immunology 2013;(11):801-806
Objective To investigate the immunomodulatory effects of heme oxygenase-1 ( HO-1) modified rat bone marrow mesenchymal stem cells ( BM-MSCs) on lymphocytes .Methods Rat BM-MSCs were cultured and identified in vitro.HO-1 gene was transduced into BM-MSCs via recombinant adenovirus (rAdV) to construct HO-1/BM MSCs, which were then co-cultured with rat spleen lymphocytes .The cellu-lar activity of lymphocytes was detected by using MTT method .The cell cycle distributions were analyzed by propidium iodide staining and flow cytometry .ELISA was used to detect the level of cytokines in the super-natant of cultured cells .The expression of lymphocyte activation markers , CD25 and CD71, were measured by using fluorescence labeled antibodies and flow cytometry .Results Compared with the control group , the lymphocytes co-cultured with HO-1/BM MSCs showed a suppressed proliferation and cell cycle progression . HO-1/BM MSCs inhibited the secretion of IL-2, TNF-αand IFN-γ, but enhanced the secretion of IL-10 in lymphocytes.Furthermore, the expression of CD25 and CD71 were also down-regulated by HO-1/BM MSCs. Conclusion Compared with BM MSCs , HO-1/BM MSCs show higher immunosuppressive effects on lym-phocytes .
2.Construction and Application of Lentiviral Vectors Expressing microRNA-203 Tough Decoy
Tao LIU ; Yuliang WANG ; Hongli SONG ; Nannan FU ; Benjuan WU ; Zhongyang SHEN
Tianjin Medical Journal 2014;(10):961-964
Objective To establish method of constructing lentiviral vectors to express microRNA (miRNA) ''tough decoy''(TuD)and to detect the effects of the TuD on cellular endogenous miRNA level and cellular phenotypes. Methods Two-step cloning strategy was utilized to first generate a universal miRNA TuD frame vector,followed by con-structing the TuD expression vector specially targeting miR-203. The package of the recombinant lentivirus was per-formed in 293T cells. Then the rat bone marrow mesenchymal stem cells(BM-MSCs)were infected by the miR-203 TuD expression lentivirus. The pSIH1-H1-copGFP vector was also packaged and the BM-MSCs infected by this lentivirus were served as control. Endogenous miR-203 level in BM-MSCs was measured by quantitative RT-PCR,and cellular vi-ability and apoptosis were detected by CCK-8 test and Annexin V-PI staining respectively. Results The miR-203 TuD expression vector was successfully constructed and inserted sequence was validated. At the 3rd,6th and 9th days after in-fected by the miR-203 TuD expression lentivirus,rat BM-MSCs exhibited a repressed endogenous miR-203 level. The miR-203 TuD also promoted viability and inhibited apoptosis of BM-MSCs. All these differences between miR-203 TuD group and control group were statistically significant. Conclusion The two-step cloning method for the construction of miRNA TuD expression vector is simple and efficient. The miRNA TuD can effectively suppress the level of the target miRNA and affect cellular phenotypes.
3.Comparison of esketamine versus dexmedetomidine in improving adverse mood after cesarean section
Dongmei ZHU ; Fengzhi LIU ; Ximing LI ; Xiaoyan ZHANG ; Benjuan LIU ; Lina ZHONG ; Peng XIA
Chinese Journal of Anesthesiology 2024;44(3):277-281
Objective:To compare esketamine versus dexmedetomidine in improving the adverse mood after cesarean section.Methods:One hundred and fourteen pregnant women undergoing elective cesarean section, aged 20-45 yr, with body mass index≤33 kg/m 2, of American Society of Anesthesiologists Physical Status classification Ⅱ or Ⅲ, were divided into 3 groups ( n=38 each) by the random number table method: esketamine group (group S), dexmedetomidine group (group D) and control group (group C). After delivery, esketamine was intravenously injected as a bolus of 0.3 mg/kg, followed by an infusion of 0.3 mg·kg -1·h -1 throughout the surgery in group S, dexmedetomidine was intravenously injected as a bolus of 0.6 μg/kg, followed by an infusion of 0.6 μg·kg -1·h -1 throughout the surgery in group D, while the equal volume of normal saline was given instead, followed by an infusion of 14 ml/h throughout the surgery in group C. Patient-controlled intravenous analgesia was performed after the end of surgery. Esketamine 50 mg, sufentanil 50 μg and ondansetron 8 mg were given in group S, dexmedetomidine 200 μg, sufentanil 50 μg and ondansetron 8 mg were given in group D, while sufentanil 50 μg and ondansetron 8 mg were given in group C. When the visual analog scale score ≥4 within 48 h after operation, flurbiprofen axidate was intravenously injected as a rescue analgesic. Self-rating Anxiety Scale (SAS) scores and Edinburgh Postnatal Depression Scale (EPDS) scores were assessed at 1 day before surgery and 2 and 7 days after surgery. Serum levels of brain-derived neurotrophic factor (BDNF) were measured by enzyme-linked immunosorbent assay at 1 day before surgery and 2 days after surgery. The effective pressing times of patient-controlled analgesia (PCA) and requirement for rescue analgesia after operation were recorded. The occurrence of adverse reactions during operation and within 48 h after operation was also recorded. Results:Compared with group C, SAS scores and EPDS scores were significantly decreased at 2 and 7 days after surgery, serum BDNF concentrations were increased at 2 days after surgery, the effective pressing times of PCA were reduced, the requirement for rescue analgesia was decreased, and the incidence of intraoperative nausea and vomiting was reduced in S and D groups ( P<0.05). Compared with group D, SAS scores and EPDS scores were significantly decreased at 7 days after surgery, the effective pressing times of PCA were reduced ( P<0.05), and no significant change was found in serum BDNF concentrations at 2 days after surgery and requirement for rescue analgesia in group S ( P>0.05). The incidence of dreaminess was significantly higher in group S than in group C and group D ( P<0.05). Conclusions:Esketamine is better than dexmedetomidine in improving the adverse mood after cesarean section.