目的:探讨穴位贴敷对妇科腹腔镜患者围术期低体温发生率的影响.方法:将2020年1月～2021年1月于我院择期手术的80例全麻妇科腹腔镜患者纳入研究,采用随机数字表法分为两组,各40例.对照组给予常规护理,观察组在对照组基础上给予穴位贴敷.比较两组患者的体温变化、术后寒战等不良反应发生率、苏醒时间、护理满意度.结果:观察组麻醉1 h后、手术结束后的中心温度明显高于对照组,苏醒时间明显短于对照组,差异有统计学意义(P＜0.05);观察组术后寒战等不良反应发生率明显低于对照组,差异有统计学意义(P＜0.05);观察组护理满意度明显优于对照组,差异有统计学意义(P＜0.05).结论:用吴茱萸等药材贴敷于穴位上可有效预防妇科腹腔镜患者围术期低体温的发生,产生积极的影响,值得临床推广.

目的:建立转染高效可调控的携带肝生长因子(hepatocyte growth factor,HGF)为目的基因的重组杆状病毒vAcrtTA2s-Ptight-HGF,并探寻其在转染骨髓间充质干细胞(bone marrow mesenchymal stem cells,BM-MSCs)后的最佳诱导浓度.方法:用转基因的方法构建重组杆状病毒v AcrtTA2s-Ptight-HGF,随后转染BM-MSCs,用ELISA和Western blot检测不同浓度强力霉素(doxycycline,DOX)诱导体外培养兔BM-MSCs分泌的HGF浓度.结果:构建了高效可调控重组杆状病毒vAcrtTA2s-Ptight-HGF,并且成功转染兔BM-MSCs,用系列浓度DOX诱导时ELISA和Western blot均检测到BM-MSCs中HGF的表达有DOX诱导剂量依赖关系并且连续7d持续表达,且实验组的HGF的表达量明显高于对照组(P＜0.001).结论:成功构建一次性转染、高效可调控的重组杆状病毒vAcrtTA2s-Ptight-HGF,并实现对HGF表达的调控,当DOX诱导浓度为1μg/ml时为其体外最佳诱导浓度.

Bone marrow-derived mesenchymal stem cells (BM-MSCs) transplantation is widely adopted for the curing of osteonecrosis of femoral head (ONFH) in recent years. Furthermore, it is known that introducing hepatocyte growth factor (HGF) into BM-MSCs will greatly improve the therapeutic effect of stem-cell therapy owing to the great angiogenic and anti-fibrotic capabilities of HGF. However, continuing overexpression of HGF in vivo may cause sarcomas, such as Kaposi's sarcoma. Aiming at enhancing the therapeutic effect and preventing the side effects of HGF-modified stem-cell transplantation on ONFH, we sought to construct a gene regulation system to control HGF expression in BM-MSCs rigorously and accurately. We selected baculovirus as the gene vector and introduced pTet-on advanced system into that. Finally, a virus vector vAc(rtTA2s-Ptight-HGF) was successfully built and delivered into BM-MSCs to regulate the accurate expression of HGF. As shown in the results, different levels of HGF expression were verified by ELISA and Western blot with different induction doses of doxycycline (DOX). There was a dose-response relationship between them, and the optimum dose of DOX to induce HGF expression in BM-MSCs in vitro was 1 µg/mL. We conclude that it is feasible to regulate HGF expression in BM-MSCs by baculovirus-mediated one-off transduction.

Objective To construct novel recombinant baculoviruses with Tet-On system and enhanced green fluorescent protein (EGFP) or hepatic growth factor (HGF) which could be regulated by different concentrations of doxycycline (DOX).Methods The recombinant plasmids pFast-Tet,pTRE-EGFP and pTRE-HGF were digested.The target fragments were collected and connected to pFast-Tet,the resultants were used to transform DH10Bac competent cells containing AcMNPV Bacmid and helper plasmid,and the Bacmid DNA were identified (named Ac-EGFP and Ac-HGF) after selection and extraction.Ac-EGFP and Ac-HGF were then transfected into bone mesenchymal stem cells (BMSCs),and the expression of EGFP and HGF were regulated by different concentrations of DOX (EGFP:0,200,500,and 1 000 ng/mL; HGF:0,10,100,500,1 000,and 1 200 ng/mL) ; EGFP expression was observed under fluorescence microscope and the level of HGF expression was detected by ELISA.Results It was verified that Tet-On system was successfully constructed in a baculovirus vector with EGFP or HGF,and they were highly transfected into BMSCs.EGFP and HGF were highly expressed when exposed to high concentrations of DOX.And the expression of EGFP and HGF were gradually decreased at low concentration or absence of DOX.Conclusion Tet-On system can be used to construct a new recombinant baculovirus vector containing EGFP or HGF and it can stably and highly transfect BMSCs; different concentrations of DOX can lead to different expression of EGFP and HGF,and they are in low background expression without DOX.