1,8-桉叶油素自微乳给药系统的制备及质量评价和细胞摄取研究

目的优化1,8-桉叶油素(1,8-cineole,1,8-Cin)自微乳给药系统(1,8-cineole self-microemulsion drug delivery system,1,8-Cin-SMEDDS)处方,对其进行表征并进行细胞摄取考察。方法通过绘制伪三元相图,确定1,8-Cin-SMEDDS有效自乳化区域,进行初步处方筛选。以粒径和载药量为指标,采用星点设计-效应面法对1,8-Cin-SMEDDS处方进行优化并验证。荧光显微镜观察高糖损伤的人脐静脉内皮细胞(HUVEC)对1,8-Cin-SMEDDS的摄取情况。结果 1,8-Cin-SMEDDS的最佳处方是大豆油(7.5%)与1,8-Cin(22.5%)为混合油相,HS15(56%)为乳化剂,乙醇(14%)为助乳化剂,滴加纯水至8m L得半透明略带蓝色乳光液体。透射电镜观察其外观呈球形液滴,激光粒度Zeta电位测定仪测得平均粒径为(131.68±1.44)nm,Zeta电位为(-10.03±1.63)m V;HPLC法测得包封率为(99.890±0.012)%,载药量为(224.750±0.028)mg/g。HUVEC细胞摄取实验结果表明,细胞对1,8-Cin-SMEDDS的摄取高于游离1,8-Cin。结论 1,8-Cin-SMEDDS制备方法简便,重复性好,所得1,8-Cin-SMEDDS外观良好,包封率高,理化性质稳定,且能促进细胞摄取。     

PFKFB4对胃癌细胞侵袭和迁移能力的影响及其作用

目的 探讨激酶PFKFB4是否参与调控胃癌细胞侵袭和迁移并对其作用机理进行初步研究。方法 使用免疫共沉淀检测与PFKFB4有相互作用的SRC家族蛋白,通过转录组测序寻找下游调控基因,利用Western blot和qRT-PCR验证下游基因与磷酸化SRC蛋白的关系,并通过Transwell方法检测PFKFB4相关通路对胃癌细胞迁移和侵袭能力的影响。结果 PFKFB4能够与SRC-2相互作用,但PFKFB4不影响SRC-2的表达,而是磷酸化SRC-2第698位的丝氨酸;SRC-2 Ser698磷酸化后,其下游调控基因LKB1的mRNA和蛋白的表达水平都受到抑制,同时胃癌细胞的迁移和侵袭能力增强。结论 PFKFB4通过磷酸化SRC-2负调控抑癌因子LKB1的表达增强胃癌细胞的迁移和侵袭能力。     

A prognostic signature based on immune-related genes for cervical squamous cell carcinoma and endocervical adenocarcinoma

Cervical squamous cell carcinoma and endocervical adenocarcinoma (CESC) is the fourth commonest female malignancy worldwide. CESC progresses in immune-microenvironment mainly composed of infiltrating immune and stromal cells. Here, we performed an integrated analysis incorporating the expression profiles from the Cancer Genome Atlas (TCGA) database and scores of immune and stromal cells calculated by Estimation of Stromal and Immune cells in Malignant Tumours using Expression data (ESTIMATE) algorithm. A two-gene signature (CD1C and CD6 genes) was established to predict the prognosis of CESC. Based on this signature, patients were divided into the high- and low-risk groups, and this signature showed good prognostic performance according to the results of Kaplan-Meier analysis and receiver operating characteristic (ROC) analysis in train set and two validation sets. A nomogram was built for evaluating the clinical applicability of this signature. In addition, based on Tumor Immune Estimation Resource (TIMER) database, 2 hub genes showed negative correlations with tumor purity and positive correlations with infiltrating levels of immune filtrating cells. What’s more, we propose new treatment strategies for the two prognostic subtypes. Low- risk patients were found presenting with a higher level of immune checkpoint molecules and showing higher immunogenicity in immunophenoscore (IPS) analysis, which indicated a better response for immunotherapy. Meanwhile, estimated by Genomics of Drug Sensitivity in Cancer (GDSC) database, the high-risk patients showed sensitive responses to five chemotherapy drugs. Finally, 10 candidate small-molecule drugs for CESC were defined. In summary, the CD1C-CD6 signature can accurately predict the prognosis of CESC.     

MiRNA let-7a对人喉鳞癌细胞增殖的抑制作用CSCD

目的探讨miRNA let-7a调控高迁移率族蛋白2(HMGA2)对人喉鳞癌细胞株TU212增殖的影响。方法合成let-7a模拟体(let-7a mimics)并采用阳离子脂质体法瞬时转染入喉癌TU212细胞,裸鼠皮下注射TU212细胞,构建裸鼠移植瘤模型。RT-qPCR和Western blot法分别检测转染后TU212细胞及移植瘤内let-7a和HMGA2的表达。结果在过表达let-7a的喉癌TU212细胞中,HMGA2的转录和翻译水平下调,细胞增殖能力降低。体外成瘤实验证实,与let-7a NC组和空白对照组相比,转染let-7a mimics的喉癌细胞TU212成瘤组织的质量和体积均显著下降;let-7a过表达的TU212成瘤组织中HMGA2的mRNA和蛋白水平均明显下调。结论let-7a能显著抑制HMGA2的mRNA和蛋白表达,进而显著抑制喉癌细胞的增殖。     

以机油为分散剂高温热解人发合成新型碳点及其生物效应研究

目的以机油为分散剂高温热解人发后发现新型纳米碳点,通过动物实验评价了该碳点的生物效应。方法利用高温热解法对人发和机油进行炭化,对炭化产物进行萃取、过滤分离和透析得到一种具有水溶性的新型物质碳点,并命名为JYRF-CDs。利用透射电子显微镜(TEM)、高分辨透射电子显微镜(HR-TEM)、紫外-可见分光光谱、傅里叶变换红外光谱、荧光光谱、X射线光电子能谱分析(XPS)等多种方法对JYRF-CDs进行表征,利用小鼠单核巨噬细胞RAW264.7细胞进行CCK-8毒性实验来评价JYRF-CDs的安全性,并通过小鼠耳肿胀实验和小鼠醋酸扭体实验对JYRF-CDs的生物效应进行评价。结果 JYRF-CDs外形为类球形,粒径均匀分布在1.8～3.6 nm,晶格间距为0.219 7 nm。细胞毒性实验结果显示,JYRF-CDs具有低毒性,动物实验结果表明JYRF-CDs具有良好的抗炎和镇痛作用。结论首次以机油为分散剂高温热解人发后发现一种全新的碳点JYRF-CDs,以JYRF-CDs为突破口,更加明确阐释以机油为分散剂高温热解人发后炭化产物具有生物效应的物质基础,为纳米类成分的研究提供了一种新方法。     

Functional beta-cells derived from umbilical cord blood mesenchymal stem cells for curing rats with streptozotocin-induced diabetes mellitus

INTRODUCTIONThis study aimed to investigate the therapeutic response to injected human umbilical cord blood mesenchymal stem cells (UCBMSCs) among albino rats with streptozotocin (STZ)-induced diabetes mellitus.METHODSControl group (GI; n = 25) rats were fed with standard rat diet. Rats with STZ-induced diabetes mellitus without (GII; n = 25) and with (GIII; n = 25) differentiated human UCBMSCs implantation were the test groups. Rats were sacrificed in Week 11 following implantation. Liver biopsies were sectioned and stained in order to highlight both the presence and function of impregnated cells in the liver tissue.RESULTSHaematoxylin and eosin-stained sections in GI and GII rats showed normal liver architecture while GIII rats showed presence of cell clusters inside the liver tissue and around the central veins. Cell clusters with blue cytoplasm were present in sections in GIII rats but absent in GI and GII rats, indicating the presence of injected differentiated human UCBMSCs. The anti-human insulin immunostaining of GIII rats showed clusters of cells within the liver parenchyma and around central veins, indicating that these cells were active and secreting insulin.CONCLUSIONUCBMSCs are proficient in differentiating into insulin-producing cells in vivo under specific conditions and, when transplanted into the liver of albino rats with STZ-induced diabetes mellitus, were able to secrete insulin and partially control the status of diabetes mellitus in rats.     

COPD患者相关炎性因子水平与其病情严重程度的相关性研究

探讨慢性阻塞性肺病(COPD)患者的可溶性髓系细胞触发受体-1(sTREM-1)、结缔组织生长因子(CTGF)、亲环素-A(CyPA)水平与其病情严重程度的相关性。ELISA检测COPD患者与健康人血清sTREM-1、CTGF、CyPA水平。结果显示,COPD患者sTREM-1、CTGF、CyPA水平高于健康人。随着肺功能分级的增加,COPD患者血清中sTREM-1、CTGF、CyPA水平也随之增加。sTREM-1、CTGF、CyPA水平与FEV1/FVC呈现显著负相关性(P＜0.05)。Logistic回归分析显示,sTREM-1、CTGF、CyPA水平是严重COPD的影响因素。结果说明,COPD患者血清sTREM-1、CTGF、CyPA水平显著升高,并且与患者肺功能指标显著相关,可用于判断COPD患者的病情严重程度。     

Therapeutic effect of mesenchymal stem cell on Hashimoto’s thyroiditis in a rat model by modulating Th17/Treg cell balance

Abstract

The autoimmune condition Hashimoto’s thyroiditis (HT) is a disease wherein lymphocytes mediate the autoimmune damage and destruction of the thyroid gland. There are currently no effective means of treating HT, with the primary strategies of thyroid hormone therapy, surgery, or immunomodulatory therapy being associated with serious risks and side effects. There is thus a clear and urgent need to identify novel treatments for HT. In this study, we utilize female SD rats induced HT to evaluated the ability of transplanted MSCs to regulate Th17/Treg interactions in a rat Hashimoto’s thyroiditis (HT) model system. The results showed that Rats in the HT model group exhibited increased thyroid autoantibody levels consistent with successful model development, whereas these levels were lower in rats treated with MSCs. There were also fewer thyroid lesions and less lymphoid infiltration of the thyroid in MSC-treated rats relative to HT model rats, as well as fewer Th17 cells and more Treg cells – an observation consistent with the cytokine analyses. All of these showed that MSCs can regulate Th17/Treg interactions in a rat Hashimoto’s thyroiditis (HT) model system. It suggested that transplanted MSCs could be a potential immunotherapy strategy for the treatment of Hashimoto’s thyroiditis.     

Mesenchymal stem cells from orthodontic premolar teeth

BackgroundConsidering the limitations in isolating Bone Marrow Mesenchymal Stem Cells (BMSCs), alternate sources of Mesenchymal Stem Cells (MSCs) are being intensely investigated. This study evaluated dental pulp MSCs (DP-MSCs) isolated from orthodontically extracted premolar teeth from a bone tissue engineering perspective.MethodsMSCs isolated from premolar teeth pulp were cultured and studied using BMSCs as the control. Flow cytometry analysis was performed for the positive and negative MSC markers. Multilineage differentiation focusing on bone regeneration was evaluated by specific growth induction culturing media and by alkaline phosphatase (ALP) activity. Data were compared by repeated measurement analysis of variance and Student's t-test at a p value <0.05.ResultsProliferation rate, population doubling time, and colony formation of DP-MSCs were significantly higher (p < 0.001) than BMSCs. More than 85% of DP-MSCs expressed CD44, CD73, CD90, CD105, and CD166. Negative reaction was found for CD11b CD33, CD34, and CD45. Positive reaction was displayed by 7.2% of cells for early MSC marker, Stro-1. Both the cell populations differentiated into adipogenic, osteogenic, and chondrogenic lineages, with adequate ALP expression.ConclusionBecause DP-MSCs from orthodontic premolars hold a neural crest/ectomesenchymal ancestry, its prudent growth characteristics and multilineage differentiation open up exciting options in craniofacial tissue engineering.     

Percentage of residual B cells after 2 weeks of rituximab treatment predicts the improvement of systemic sclerosis-associated interstitial lung disease

The benefit of rituximab (RTX) for systemic sclerosis-associated interstitial lung disease (SSc-ILD) has been shown in previous clinical trials. However, predictors of RTX efficacy have not been clarified. We investigated whether B-cell responsiveness to RTX is related to therapeutic effect. Ten SSc-ILD patients treated with RTX in an independent clinical trial (Japan Registry of Clinical Trials, jRCTs031180373) were included in this analysis. Peripheral B-cell counts were examined retrospectively before RTX administration (baseline) and at 2, 4, 12, and 24 weeks after the first RTX administration, along with percent-predicted forced vital capacity (%FVC) before and 24 weeks after RTX treatment. Relative to baseline, the percentage of residual peripheral blood B cells at 2 weeks after RTX was negatively correlated with the %FVC improvement at the 24-week assessment (r = ?0.41, p = 0.04). In the subgroup with less than 5% B-cell persistence at week 2, %FVC at the 24-week assessment was significantly improved compared to baseline (p = 0.02). In another subgroup with more than 5% residual B cells, %FVC was not significantly different after 24 weeks compared to baseline (p = 0.41). In conclusion, the removal rate of B cells after 2 weeks of RTX treatment may be a useful surrogate marker of subsequent SSc-ILD improvement.