Different roles of IL-15 from IL-2 in differentiation and activation of human CD3+CD56+ NKT-like cells from cord blood in long term culture

The abundantly available source of stem cells and the low incidence of graft-versus-host disease (GVHD) made cord blood an attractive alternative source of hematopoietic stem cells for transplantation. Besides T cell and NK cell, NKT cell played an important role in low incidence of GVHD during allogeneic transplantation. IL-2 and IL-15 can stimulate T cell and NK cell proliferation, survival and activation in vitro. But they exhibited different effects on the GVHD during allogeneic transplantation. In this study, we explored the different effects of exogenous IL-2 and IL-15 on the expansion of CD3+CD56+ NKT-like cells by in vitro long term culture of cord blood mononuclear cells (CBMCs). The results showed that CD3+CD56+ NKT-like cells were derived from CD34-CD56- CBMCs and IL-2 improved CD3+CD56+ NKT-like cell expansion more strongly than IL-15. Interestingly, CD3+CD56+ NKT-like cells from IL-15-cocultured CBMCs had significantly lower apoptotic frequency and higher levels of activation markers (CD161, CD25, and IFN-gamma) than those from IL-2-cocultured CBMCs. The anti-apoptotic and activating effects of IL-15 on CD3+CD56+ NKT-like cells from CBMCs might possibly explain the pathogenic role of IL-15 in GVHD during allogeneic transplantation.     

Lyophilized Cheliensisin A submicron emulsion for intravenous injection: characterization, in vitro and in vivo antitumor effect

Growing attentions have been focused on natural antitumor drugs. Recently, a novel and potent antitumor drug Cheliensisin A (GC-51) with broad-spectrum efficiency has been developed. However, due to its poor water solubility and chemical instability, choosing the appropriate dosage form is of great significance. This study aimed at developing a lyophilized submicron emulsion for GC-51 and further improving the therapeutic index of the drug. The resultant lyophilized GC-51 submicron emulsion was much more stable than its solution, which can be stored for years without significant change on physicochemical properties. And its solubility was increased from 6.74 ± 0.14 to 2.00 ± 0.10 mg mL⁻¹. The 50% inhibitory concentration IC₅₀ values were calculated from growth curves by MTT assay on various tumor cell lines. Compared with the IC₅₀ of GC-51 crude drug, that of lyophilized GC-51 submicron emulsion decreased from 24.04 ± 1.97 to 8.23 ± 1.84 μg mL⁻¹ on HepG2, and from 31.08 ± 2.56 to 10.85 ± 2.09 μg mL⁻¹ on CT-26, from 17.90 ± 1.83 to 7.49 ± 1.87 μg mL⁻¹ on HeLa and from 16.38 ± 2.41 to 10.13 ± 2.12 μg mL⁻¹ on A549, respectively. In the time-dependent assay of tumor cell viability, lyophilized GC-51 submicron emulsion exhibited significantly lower inhibition rate in the initial action times, but increased gradually afterwards. That means lyophilized submicron emulsion as the vector for GC-51 had some protective and delayed release effect. Further, the in vivo therapeutic efficacy was measured in pulmonary metastasis of colon cancer-bearing BALB/c mice model. An obvious enhanced antitumor activity was observed after administration of lyophilized GC-51 submicron emulsion (P < 0.05), which increased from 22.78 ± 3.5 to 41.42 ± 4.2% compared with GC-51 injection. And the life span of tumor-bearing mice in lyophilized GC-51 submicron emulsion group was significantly longer than that of the mice in GC-51 injection and normal saline groups. Compared with crude drug, the lyophilized GC-51 submicron emulsion showed a significantly higher antitumor efficiency both in vivo and in vitro, suggesting a potential application in tumor chemotherapy.     

Suppression of ClC-3 channel expression reduces migration of nasopharyngeal carcinoma cells

Recent studies suggest that chloride (Cl-) channels regulate tumor cell migration. In this report, we have used antisense oligonucleotides specific for ClC-3, the most likely molecular candidate for the volume-activated Cl- channel, to investigate the role of ClC-3 in the migration of nasopharyngeal carcinoma cells (CNE-2Z) in vitro. We found that suppression of ClC-3 expression inhibited the migration of CNE-2Z cells in a concentration-dependent manner. Whole-cell patch-clamp recordings and image analysis further demonstrated that ClC-3 suppression inhibited the volume-activated Cl- current (I(Cl,vol)) and regulatory volume decrease (RVD) of CNE-2Z cells. The expression of ClC-3 positively correlated with cell migration, I(Cl,vol) and RVD. These results strongly suggest that ClC-3 is a component or regulator of the volume-activated Cl- channel. ClC-3 may regulate CNE-2Z cell migration by modulating cell volume. ClC-3 may be a new target for cancer therapies.     

PEGylated nanostructured lipid carriers loaded with 10-hydroxycamptothecin: an efficient carrier with enhanced anti-tumour effects against lung cancer

Most drugs do not have the pharmacokinetic features required for optimal pulmonary delivery. In this study, we developed PEGylated nanostructured lipid carriers (PEG-NLCs) to improve the delivery of anti-tumour agents to lung tumours. PEG-40 NLCs modified with PEG-40 stearate (molecular weight 2000 Da), PEG-100 NLCs modified with PEG-100 stearate (molecular weight 5000 Da) and NLCs without PEG modification were prepared by melt-emulsification and homogenization, and were loaded with 10-hydroxycamptothecin (HCPT). They were investigated in terms of physiological characteristics, biodistribution, cellular uptake, and anti-tumour effect in-vivo. PEG-NLCs exhibited regular morphology, with a spherical shape. The particle size (measured by laser diffraction) was approximately 100 nm. Encapsulation in PEG-NLCs protected the active lactone form of HCPT compared with HCPT solution after incubation with plasma. In biodistribution studies, PEG-NLCs, especially PEG-40 NLCs, had longer circulation time and decreased uptake by the reticuloendothelial system (RES) compared with unmodified NLCs. PEG-NLCs accumulated in the lungs after i.v. injection in mice. PEG-NLCs showed enhanced cellular uptake by human lung adenocarcinoma epithelial A549 cells. In-vivo experiments indicated that PEG-NLCs loaded with HCPT have superior efficacy against A549 lung cancer compared with HCPT solution and NLCs. These results suggest that PEG-NLCs is a promising delivery system for HCPT in the treatment of lung cancer.     

3 340例住院患儿血清25-羟维生素D水平的临床观察

摘 要 目的：了解3 340例住院患儿血清25 羟维生素D[25 (OH)D]的基线水平和儿科常见疾病谱、不同年龄段、不同性别及季节的25-(OH)D水平,并分析其与临床指标之间的相关性。 方法: 随机选取3 340例住院患儿,ELISA方法检测血清25 (OH)D的水平,并用Pearson相关分析探讨25-(OH)D水平与患儿临床指标如肝功能、心肌酶谱、免疫球蛋白、T淋巴细胞亚型、甲状腺功能的相关性。结果: 3 340例患儿中男1 850例,女1 490例,平均血清25-(OH)D水平为（33.00±13.42）ng.m1^-1。新生儿疾病患儿血清25 (OH)D水平最低,其次是原发性肾病综合征、过敏性紫癜及幼年类风湿性关节炎患儿。足月儿较早产儿血清25-(OH)D水平高。除新生儿外,随年龄增加,患儿血清25-(OH)D水平逐渐降低,不足百分比逐渐增加。不同年龄组男性、女性25-(OH)D水平无显著差异。住院患儿夏季25-(OH)D水平最高。25-(OH)D与体质指数(BMI)、ALT、AST、磷酸肌酸激酶（CK）、磷酸肌酸激酶同工酶（CK-MB）、乳酸脱氢酶（LDH）、游离T4(FT4)、游离T3(F3)呈正相关,与AKP、免疫球蛋白E（IgE）、免疫球蛋白M（IgM）呈负相关。结论:住院患儿维生素D不足发生率较高,不同疾病其不足程度不同,血清25 (OH)D水平与BMI、过敏、心肌受损、甲状腺功能可能有一定的相关性。     

天麻钩藤饮联合丁苯酞治疗急性脑梗死患者的临床效果

目的:探讨天麻钩藤饮联合丁苯酞治疗急性脑梗死(ACI)的临床效果。方法:选取2018年6月至2019年4月六安市中医院收治的ACI患者50例作为研究对象,按照随机数字表法分为对照组和观察组,每组25例。对照组给予常规治疗,观察组在此基础上给予天麻钩藤饮联合丁苯酞治疗。统计2组患者临床治疗有效率;比较2组患者治疗前后血清丙二醛(MDA)、超氧化物歧化酶(SOD)、谷胱甘肽过氧化酶(GSH-PX)水平、氧化型低密度脂蛋白(ox-LDL)水平变化情况;观察2组患者治疗前后美国国立卫生研究院卒中量表(NIHSS)和改良Rar Ikin量表(MRS)评分、日常生活活动能力(BI)评分情况;结果:治疗后观察组NIHSS、MRS评分与MDA、ox-LDL水平均显著低于治疗前,且NIHSS、MRS评分与MDA水平均明显低于对照组,差异有统计学意义(P 0. 05);治疗后观察组BI评分和SOD、GSH-PX水平均明显高于治疗前与对照组,差异有统计学意义(P 0. 05);治疗后观察组临床治疗有效率明显高于对照组,差异有统计学意义(P 0. 05)。结论:天麻钩藤饮联合丁苯酞治疗可有效改善ACI患者的临床症状,增强机体清除自由基的能力,减轻氧化刺激反应,减轻脑缺血损伤,提高临床治疗有效率,且疗效优于常规治疗。     

Salubrinal Suppresses IL-17-Induced Upregulation of MMP-13 and Extracellular Matrix Degradation Through the NF-kB Pathway in Human Nucleus Pulposus Cells

Matrix metalloproteinase 13 (MMP-13) plays an important role in the process of pro-inflammatory cytokine-induced intervertebral disc degeneration (IDD). This study examined the effect of IL-17 on the regulation of MMP-13 and the extracellular matrix (ECM) in the intervertebral disc (IVD). We then examined whether salubrinal, a known inhibitor of eIF2α dephosphorylation, inhibited the IL-17-induced changes mentioned above. Furthermore, we demonstrated a potential therapeutic role for salubrinal in alleviating the chronic inflammatory-dependent degenerative state commonly observed in IDD. After inflammatory distress with IL-17, RT-PCR and western blot were employed to investigate the expression of MMP-13, collagen type II (COL2A1), collagen type I (COL1A1), and aggrecan (ACAN) in nucleus pulpous (NP) tissue. Activation of the NF-kB pathway was measured by western blot and immunocytochemistry following IL-17 treatment. We also examine the level of eIF2α phosphorylation after IL-17 treatment with or without salubrinal. Then, we investigated interactions of the NF-kB pathway to eIF2α phosphorylation. Moreover, we employed salubrinal and a specific inhibitor of NF-kB (BAY11-7082) to evaluate their effects on IL-17-driven regulation of MMP-13 and the ECM, as well as on the activation of NF-kB. The results showed that IL-17 increased the production of MMP-13 and decreased expression of COL2A1 and ACAN via the NF-kB pathway. Either IL-17 or salubrinal increased the level of eIF2α phosphorylation, but the effects of BAY11-7082 on the level of p-eIF2α were not detectable. BAY11-7082 and salubrinal significantly suppressed IL-17-driven intervertebral disc degeneration. Furthermore, salubrinal produced stronger effects than BAY11-7082. These results imply the potential involvement of IL-17 in IDD through activation of NF-kB signaling, which successively upregulated the expression of MMP-13 and led to the degradation of the ECM. Furthermore, salubrinal can inhibit this process through inhibition of NF-kB activation that is not directly linked to eIF2α phosphorylation, suggesting a potential therapeutic role in IDD.     

CT静脉成像在布加综合征诊疗中的应用

目的：探讨CT静脉成像（CTV）技术在布加综合征（BCS）诊疗过程中的应用价值。方法对2012年1月至2014年1月该院收治的58例BCS患者进行CTV检查,对下腔静脉、肝静脉、门静脉及侧支血管进行重建分析。结果所有患者均完成CTV检查,被明确诊断为BCS。其中下腔静脉隔膜型19例,短段病变型15例,长段病变型8例,肝静脉型9例,合并血栓形成的7例。所有患者均见到侧支血管不同程度的代偿扩张,肝脏硬化及脾脏肿大等表现。结论 CTV技术的应用对于BCS的诊疗具有图像清晰、直观、全面,能够显示病变血管部位、性质及长度的优点,还能观察侧支血管扩张及肝硬化程度。