【摘要】 目的 探讨LIM矿化蛋白（LIM mineralization protein，LMP）-1和LMP-3双基因共转染骨髓间充质干细胞（bone mesenchymal stem cells，BMSC）的表达情况。 方法 采用人工设计合成人LMP-1和LMP-3基因片段，分别与质粒pEGFP-N2连接，经酶切、测序鉴定后。分离培养新西兰兔BMSC，用脂质体包裹转染BMSC，按转染情况分为5组:未转染组（A组）、转染空载体组（B组）、转染LMP-1基因组（C组）、转染LMP-3基因组（D组）、LMP-1与LMP-3双基因共转染组（E组）。采用实时聚合酶链反应（real-time polymerase chain reaction，RT-PCR）和蛋白质印迹法检测LMP-1和LMP-3的表达。 结果 酶切及测序表明真核表达质粒pEGFP-N2-LMP-1和pEGFP-N2-LMP-3构建成功。E组可同时较高水平表达LMP-1和LMP-3分子。对RT-PCR及蛋白质印迹法检测结果行灰度值测量并行统计学分析显示:LMP-1 mRNA及蛋白水平的表达，5组间差异有统计学意义（Plt;0.05），但E组与C组的差异无统计学意义（Pgt;0.05）；LMP-3 mRNA及蛋白水平的表达，5组间差异有统计学意义（Plt;0.05），且E组与D组差异也有统计学意义（Plt;0.05）。 结论 双基因共转染的BMSC能在体外同时表达LMP-1与LMP-3，为基因修复骨缺损带来新思路。【Abstract】 Objective To study the expression of LIM mineralization protein (LMP)-1 and LMP-3 genes after cotransfecting them into bone mesenchymal stem cells (BMSC) of rabbit in vitro. Methods Fragments of LMP-1 gene and LMP-3 gene were gained through artificial synthesis, and were constructed respectively into the plasmid vector pEGFP-N2. The inserted target genes in plasmid were verified by nucleotide sequencing and enzymes. The plasmids carrying LMP-1 and LMP-3 genes were cotransfected into chondrocytes by liposome method. According to the transfected situation, the BMSC were divided into 5 groups: the non-transfected group (Group A), the group transfected by empty vector (Group B), the group transfected by LMP-1 (Group C), the group transfected by LMP-3 (Group D) and the group transfected by both LMP-1 and LMP-3 (Group E). The expressions of LMP-1 and LMP-3 were detected by RT-PCR and western bloting technique. Results The plasmid pEGFP-N2-LMP-1 and pEGFP-N2-LMP-1 were obtained successfully by cloning technique and verified by nucleotide sequencing and enzymes. The LMP-1 and LMP-3 molecules were both expressed at a high level in Group E. The results of RT-PCR and western bloting were measured with the grey value. For the expression of LMP-1 mRNA and protein of LMP-1, the differences between groups A, B and groups C, D, E were significant (P＜0.05), while the difference between groups C and E was not significant (P＞0.05); For the expression of LMP-3 mRNA and protein of LMP-3, the differences between groups A, B and groups C, D, E were significant (P＜0.05), and the difference between groups D and E was also significant（P＜0.05). Conclusion LMP-1 and LMP-3 genes can be expressed effectively after being cotransfected into BMSC, which provides a basis for gene therapy for treating bone defects.
Objective To explore the prognostic value of red cell volume distribution width (RDW) for hematological malignancies. Methods PubMed, Embase, Web of Science, Cochrane Library, China National Knowledge Infrastructure, Wanfang, Chongqing VIP, and SinoMed were searched for related literatures on myelodysplastic syndrome, leukemia and other hematological malignancies and pretreatment RDW from the establishment of databases to April 5, 2022. The main statistical indicators were Hazard ratio (HR) and its 95% confidence interval (CI). Stata 12.0 SE software was used for analysis, and Q test was used to evaluate literature heterogeneity. Subgroup pooled analysis was used to evaluate the prognostic value of RDW. Results A total of 7 articles were included, with a total of 804 patients. A fixed-effect model was selected for meta-analysis, and the results showed that patients with elevated pretreatment RDW had worse overall survival [HR=2.91, 95%CI (2.01, 4.22), I2=0%, P=0.714]. The results of subgroup analysis for different types of diseases showed that in myelodysplastic syndrome group [HR=2.61, 95%CI (1.28, 5.31), I2=22.0%, P=0.258)], chronic myeloid leukemia group [HR= 3.24, 95%CI (1.91, 5.51), I2=0%, P=0.546], and adult T-cell leukemia/lymphoma group [HR=2.64, 95%CI (1.22, 5.70)], the overall survival rate of patients with elevated pretreatment RDW were worse. Sensitivity analysis showed that the study was stable and there was no heterogeneity in the overall study result.Conclusion Elevated pretreatment RDW is associated with overall survival and can be used as an indicator for evaluating the prognosis of hematological malignancies, but large sample studies are still needed to determine the best predictive cutoff for various diseases.
At present, the incidence of Parkinson’s disease (PD) is gradually increasing. This seriously affects the quality of life of patients, and the burden of diagnosis and treatment is increasing. However, the disease is difficult to intervene in early stage as early monitoring means are limited. Aiming to find an effective biomarker of PD, this work extracted correlation between each pair of electroencephalogram (EEG) channels for each frequency band using weighted symbolic mutual information and k-means clustering. The results showed that State1 of Beta frequency band (P = 0.034) and State5 of Gamma frequency band (P = 0.010) could be used to differentiate health controls and off-medication Parkinson’s disease patients. These findings indicated that there were significant differences in the resting channel-wise correlation states between PD patients and healthy subjects. However, no significant differences were found between PD-on and PD-off patients, and between PD-on patients and healthy controls. This may provide a clinical diagnosis reference for Parkinson’s disease.
Adverse drug reactions (ADR) are one important type of safety data for investigational products and post-market drugs. Standardized coding is beneficial to normative ADR analysis and reporting. However, terms peculiar to traditional Chinese medicine (TCM) are not included in the widely used international adverse reaction terminologies. This paper briefly introduced the differences of World Health Organization adverse reaction terminology (WHOART) and medical dictionary for regulatory activities (MedDRA) which was developed by the International Council for Harmonization of Technical Requirements for Pharmaceuticals for Human Use (ICH). Based on the existing development of TCM terminologies for coding ADRs, basic path for developing internationally recognized TCM terminologies was proposed in this paper.