Assessment of the Safety and Efficacy of an Attenuated Live Vaccine Based on Highly Pathogenic Porcine Reproductive and Respiratory Syndrome Virus

The safety and efficacy of the JXA1-R vaccine, an attenuated strain of highly pathogenic porcine reproductive and respiratory syndrome virus (HP-PRRSV), were examined using an intramuscular challenge model in piglets. The JXA1-R vaccine was obtained by passing HP-PRRSV JXA1 through Marc-145 cells (82nd passage). Genomic sequence comparisons showed that strain JXA1-R and its parental strain, JXA1, differ by 47 amino acids, and most of these differences are scattered throughout the PRRSV genome. Four-week-old PRRSV-free piglets were inoculated intramuscularly with JXA1-R vaccine (10^3.0, 10^4.0, 10^5.0, 10^6.0, and 10^7.0 50% tissue culture infective doses [TCID₅₀]/ml for groups 1 to 5, respectively) and then challenged intramuscularly with the 5th passage virus of JXA1 virus (JXA1-F5, 3 ml × 10^4.5 TCID₅₀/ml) 28 days after inoculation. The humoral immune response, swine growth, clinical signs, and differential organ lesions were monitored. The results showed that all vaccinated piglets had a perceptible humoral immune response to vaccination after day 7, which then promptly increased, almost reaching the maximum sample/positive (S/P) ratio value at 28 days postimmunization. Viremia detection indicated that the viral replication levels of the challenge virus in the immunized groups (immunization doses ≥10^4.0/ml) were significantly lower than that of the virus-challenged unvaccinated control group. Piglets in groups 2 to 5 were effectively protected against lethal HP-PRRSV infection and did not show any obvious changes in body temperature or clinical signs of disease at any point during the experiment. However, two of five piglets in group 1 showed mild pathological lesions and transitory high fever. These results suggest that JXA1-R (TCID₅₀/ml ≥10^4.0) is sufficiently attenuated and can provide effective protection against the lethal wild-type HP-PRRSV.     

内镜下射频消融术治疗胃低级别上皮内瘤变的临床研究

目的通过临床大样本研究,进一步明确内镜下射频消融术(RFA)治疗胃低级别上皮内瘤变(LGIN)的有效性及安全性。 方法回顾性选择2014年10月至2019年12月经解放军总医院消化内镜中心术前筛查的175例患者证实为胃LGIN的255例病变纳入研究,并进行内镜下RFA,关注其围手术期并发症发生情况,术后采用Wong-Baker面部表情量表进行疼痛评分,并追踪其复查随访结果。 结果255例病变均成功完成内镜下RFA,术中无明显并发症发生;术后3个月、6个月、1年、2年、3年的治愈率分别为91.3%、90.8%、89.4%、88.2%、86.5%,术后腹痛为主要并发症。 结论内镜下RFA是治疗胃LGIN的一种安全有效、操作简便、可门诊治疗的新方法,具有良好的临床应用前景。     

白细胞介素27在结核性胸腔积液和恶性胸腔积液鉴别诊断中的价值

目的研究胸腔积液中白细胞介素27(IL-27)浓度对于结核性胸腔积液(TPE)和恶性胸腔积液(MPE)的鉴别诊断价值.方法纳入2015~2018年收住北京朝阳医院诊断为TPE或MPE的患者,通过酶联免疫吸附法测定胸腔积液及血浆中IL-27浓度.通过受试者工作特征曲线获取胸腔积液中IL-27的浓度、胸腔积液与血浆IL-27浓度的差值和比值的诊断价值.结果143例患者中TPE组78例,MPE组65例.TPE患者中胸腔积液IL-27浓度显著高于MPE患者,胸腔积液IL-27诊断TPE的诊断界值为499.71 ng/L,敏感度为93.6%,特异度为95.4%,曲线下面积为0.952(95%CI:0.913~0.991),Z统计分析显示,胸腔积液与血浆IL-27浓度差值和比值的诊断价值不高于胸腔积液IL-27浓度.结论胸腔积液IL-27浓度是鉴别TPE和MPE的理想生物标志物.     

Branched polyethyleneimine-assisted 3-carboxybenzoboroxole improved Wulff-type boronic acid functionalized magnetic nanoparticles for the specific capture of cis-diol-containing flavonoids under neutral conditions

Flavonoids have shown a variety of biological activities such as antimicrobial, antibacterial, antifungal, antiviral, antiinflammatory, antitumor, antiatherogenic, and antihyperglycemic activities. A lot of important flavonoids contain cis-diols such as rutin (Ru), quercetin (Qu), luteolin (Lu), myricetin (Myr) and baicalein (Ba) and so on. It is necessary to establish a simple, low-cost and efficient purification method for cis-diol-containing flavonoids from plant extracts. Boronate affinity materials are able to reversibly bind the cis-diols via boronic acids by forming a five- or six-membered boronic cyclic ester in aqueous media. However, conventional boronate affinity materials have to be used in alkaline media, which can lead to the oxidation of cis-diols in compounds. In this study, the polyethyleneimine (PEI)-assisted 3-carboxybenzoboroxole-functionalized magnetic nanoparticles (MNPs) were prepared to achieve efficient capture of cis-diol-containing flavonoids under neutral conditions. Branched PEI was applied as a scaffold to amplify the number of boronic acid moieties, while 3-carboxybenzoboroxole, exhibiting high affinity and excellent water solubility toward flavonoids, was used as an affinity ligand. The prepared boronate affinity MNPs exhibited high binding capacity and fast binding kinetics (equilibrium in 3 min) under neutral conditions. In addition, the obtained boronate affinity MNPs exhibited high binding affinity (K_d ≈ 10⁻⁴ M), low binding pH (pH ≥ 6.0) and tolerance of the interference to abundant sugars.

Flavonoids have shown a variety of biological activities such as antimicrobial, antibacterial, antifungal, antiviral, antiinflammatory, antitumor, antiatherogenic, and antihyperglycemic activities.     

Quantitative phosphoproteomic analysis reveals system-wide signaling pathways downstream of SDF-1/CXCR4 in breast cancer stem cells

Breast cancer is the leading cause of cancer-related mortality in women worldwide, with an estimated 1.7 million new cases and 522,000 deaths around the world in 2012 alone. Cancer stem cells (CSCs) are essential for tumor reoccurrence and metastasis which is the major source of cancer lethality. G protein-coupled receptor chemokine (C-X-C motif) receptor 4 (CXCR4) is critical for tumor metastasis. However, stromal cell-derived factor 1 (SDF-1)/CXCR4–mediated signaling pathways in breast CSCs are largely unknown. Using isotope reductive dimethylation and large-scale MS-based quantitative phosphoproteome analysis, we examined protein phosphorylation induced by SDF-1/CXCR4 signaling in breast CSCs. We quantified more than 11,000 phosphorylation sites in 2,500 phosphoproteins. Of these phosphosites, 87% were statistically unchanged in abundance in response to SDF-1/CXCR4 stimulation. In contrast, 545 phosphosites in 266 phosphoproteins were significantly increased, whereas 113 phosphosites in 74 phosphoproteins were significantly decreased. SDF-1/CXCR4 increases phosphorylation in 60 cell migration- and invasion-related proteins, of them 43 (>70%) phosphoproteins are unrecognized. In addition, SDF-1/CXCR4 upregulates the phosphorylation of 44 previously uncharacterized kinases, 8 phosphatases, and 1 endogenous phosphatase inhibitor. Using computational approaches, we performed system-based analyses examining SDF-1/CXCR4–mediated phosphoproteome, including construction of kinase–substrate network and feedback regulation loops downstream of SDF-1/CXCR4 signaling in breast CSCs. We identified a previously unidentified SDF-1/CXCR4-PKA-MAP2K2-ERK signaling pathway and demonstrated the feedback regulation on MEK, ERK1/2, δ-catenin, and PPP1Cα in SDF-1/CXCR4 signaling in breast CSCs. This study gives a system-wide view of phosphorylation events downstream of SDF-1/CXCR4 signaling in breast CSCs, providing a resource for the study of CSC-targeted cancer therapy.Breast cancer is the most common cancer in women, with an estimated 1.7 million new cases and 522,000 deaths around the world in 2012 alone. Tumor metastasis is the major source of cancer lethality. Cancer stem cells (CSCs) are small-percentage subpopulation within tumors, which are essential for tumor reoccurrence and metastasis (1). G protein-coupled receptor CXCR4 is critical for tumor growth and metastasis and plays important roles in CSC migration, invasion, and proliferation (2). Chemokine stromal cell-derived factor 1 (SDF-1) (CXCL-12) binds to chemokine (C-X-C motif) receptor 4 (CXCR4) and induces SDF-1/CXCR4 signaling. SDF-1 or CXCR4 knockout mice are embryonic lethal. SDF-1 and CXCR4 are vital for tumor angiogenesis and metastasis (3). The large-scale signal transduction and the feedback regulation downstream of SDF-1/CXCR4 signaling in breast CSCs are unknown but critical to understanding the cellular physiology of breast tumor regrowth and metastasis.Protein phosphorylation and dephosphorylation are essential for cellular signal processing (4). Dynamic regulation of reversible, site-specific protein phosphorylation is critical to the signaling networks that regulate CSC self-renewal, differentiation, and metastasis. Protein-reversible phosphorylation has been extensively analyzed in examining one or a few protein phosphorylation events that affect CSC signaling (1). However, the phosphoproteome composed by protein kinase-driven and phosphatase-regulated signaling networks largely controls CSC fate. Therefore, large-scale analysis of differentially regulated protein phosphorylation is central to understanding complex cellular events, such as CSC maintenance and dissemination.To unveil the signal transduction downstream of SDF-1/CXCR4 signaling in CSCs, in this study we have carried out isotope reductive dimethylation and large-scale liquid chromatography tandem mass spectrometry (LC-MS/MS)-based phosphoproteomic profiling and quantification in human breast CSCs upon SDF-1/CXCR4 stimulation. The phosphorylation events presented here include SDF-1/CXCR4–mediated phosphorylation sites in several key kinases and phosphatases, and several important signaling pathways in breast CSCs.     

平喘益气颗粒联合沙美特罗替卡松气雾剂治疗慢性阻塞性肺疾病稳定期的临床研究

目的探讨平喘益气颗粒联合沙美特罗替卡松气雾剂治疗慢性阻塞性肺疾病稳定期的临床疗效。方法选取2017年6月—2018年6月在天津市黄河医院治疗的94例慢性阻塞性肺疾病稳定期患者为研究对象,将患者按随机数表法分为对照组和治疗组,每组各47例。对照组吸入沙美特罗替卡松吸入气雾剂,2揿/次,2次/d。治疗组在对照组基础上口服平喘益气颗粒,2袋/次,3次/d。两组患者连续治疗3个月。观察两组的临床疗效,比较两组的肺功能指标、SGRQ评分、6 min步行距离、免疫功能、炎症因子。结果治疗后,对照组和治疗组的总有效率分别为76.60%、91.49%,两组比较差异有统计学意义(P0.05)。治疗后,两组第1秒用力呼气容积(FEV1)、用力肺活量(FVC)、FEV1/FVC均显著提高,同组治疗前后比较差异有统计学意义(P0.05);且治疗后治疗组FEV1、FVC、FEV1/FVC明显高于对照组,差异有统计学意义(P0.05)。治疗后,两组圣乔治呼吸问卷(SGRQ)评分显著降低,6 min步行距离显著增加,同组治疗前后比较差异有统计学意义(P0.05);且治疗后治疗组SGRQ评分低于对照组,6 min步行距离长于对照组,差异有统计学意义(P0.05)。治疗后,治疗组CD3~+、CD4~+、CD4~+/CD8~+明显高于治疗前,且治疗组CD3~+、CD4~+、CD4~+/CD8~+明显高于对照组,差异有统计学意义(P0.05)。治疗后,两组肿瘤坏死因子-α(TNF-α)、白细胞介素-8(IL-8)水平显著下降,同组治疗前后比较差异有统计学意义(P0.05);且治疗后治疗组炎症因子水平明显低于对照组,差异有统计学意义(P0.05)。结论平喘益气颗粒联合沙美特罗替卡松气雾剂治疗慢性阻塞性肺疾病稳定期具有较好的临床疗效,可改善患者临床症状及肺功能,调节免疫功能及相关炎症因子,提高生活质量,具有一定的临床推广应用价值。