目的 通过采用静脉注射α-半乳糖基神经酰胺(α-GalCer),观察其对小鼠肝脏恒定性自然杀伤T细胞(iNKT)频率及亚群的影响.方法 研究时间2021年11月至2022年2月,选取C57BL/6J健康小鼠,采用随机数字表法分为观察组和对照组,观察组静脉注射α-GalCer,对照组静脉注射生理盐水,在注射后的第2、4、6、8、10天取小鼠肝脏,采用流式细胞术检测小鼠肝脏的iNKT细胞及亚群变化情况;采用酶联免疫吸附测定(ELISA)检测外周血血清中白细胞介素-4(IL-4),干扰素-γ(IFN-γ),白细胞介素-17A(IL-17A)的表达水平.结果 与对照组相比,静脉注射α-GalCer可引起肝脏iNKT细胞频率及亚群频率变化,且差异有统计学意义(P<0.05).观察组肝脏iNKT细胞频率明显升高,但随时间呈逐渐降低趋势,自第6天恢复至与正常水平相当[观察组第2、4、6、8、10天分别为(20.87±2.15)％、(5.61±0.51)％、(2.65±0.26)％、(2.62±0.64)％、(2.51±0.11)％,对照组分别为(2.72±0.19)％、(2.72±0.19)％、(2.93±0.24)％、(3.06±0.45)％、(3.14±0.35)％];注射α-GalCer后,iNKT1亚群频率低于对照组,于第10天恢复至正常水平[观察组第2、4、6、8、10天分别为(1.04±0.37)％、(0.48±0.12)％、(1.99±1.04)％、(0.60±0.11)％、(3.25±0.53)％,对照组分别为(3.68±1.02)％、(2.74±0.21)％、(2.81±0.33)％、(2.93±0.29)％、(3.12±0.11)％];iNKT2亚群频率高于对照组,后逐渐降低[观察组第2、4、6、8、10天分别为(75.13±3.48)％、(33.83±9.08)％、(17.20±5.37)％、(12.00±1.99)％、(6.26±1.28)％,对照组分别为(38.03±1.70)％、(40.13±3.20)％、(40.16±3.15)％、(38.03±1.70)％、(36.37±1.18)％];iNKT1亚群频率总体上低于iNKT2亚群;iNKT17亚群频率低于对照组,第4天亚群频率最高,后又降至正常水平以下;与对照组相比,血清中IL-4水平在注射第2天无明显变化(P>0.05),注射后第6天,IL-4水平明显降低(P<0.05);IFN-γ水平自注射α-GalCer后明显升高,第6天又降低(P<0.05),IL-17A水平均低于正常水平(P<0.05).结论 静脉注射α-GalCer可诱导小鼠肝脏iNKT细胞的增殖,影响其亚群的分化.

Epigenetic regulation affects the development and differentiation of iNKT cells. Our previous study found that the number of iNKT cells in thymus of RA mice was reduced and the ratio of subsets was unbalanced, but the related mechanism remains unclear. We adopted an adoptive infusion of iNKT2 cells with specific phenotypes and functions to RA mice and used the α-Galcer treatment group as control. The findings revealed that: 1. Adoptive treatment of iNKT cells decreased the proportion of iNKT1 and iNKT17 subsets in the thymus of RA mice, and increased the proportion of iNKT2 subsets. 2. Following treatment with iNKT cells, the expression of PLZF in thymus DP T cells was increased whereas the expression of T-bet in thymus iNKT cells was decreased in RA mice. 3. Adoptive therapy reduced the modification levels of H3Kb7me3 and H3K4me3 in the promoter regions of Zbtb16 (encoding PLZF) and Tbx21 (encoding T-bet) gene in thymus DP T cells and iNKT cells, and the reduction of H3K4me3 was particularly significant in the cell treatment group. Furthermore, adoptive therapy also upregulated the expression of UTX (histone demethylase) in thymus lymphocytes of RA mice. As a result, it is hypothesized that adoptive therapy of iNKT2 cells may affect the level of histone methylation in the promoter region of important transcription factor genes for iNKT development and differentiation, thereby directly or indirectly correcting the imbalance of iNKT subsets in the thymus of RA mice. These findings offer a fresh rationale and concept for the management of RA that targets.

目的 探讨腹腔注射 α-半乳糖苷神经酰胺(α-GalCer)对小鼠肺恒定自然杀伤 T细胞(iNKT)、细胞亚群及外周血细胞因子的影响,从而为治疗相关肺部疾病提供基础研究数据.方法 60只6～8周龄雄性C57BL/6 小鼠,采用随机数字表法分为α-GalCer干预组和正常对照组,每组30只,选择不同时间点观察干预组与对照组小鼠肺iNKT 细胞、亚群及外周血细胞因子的变化.结果 组间比较发现,与对照组(3.49±0.25)%相比,腹腔注射α-GalCer可引起肺脏iNKT细胞频率及亚群的变化,差异有统计学意义(P<0.05).α-GalCer干预组肺iNKT细胞频率前2 d(6.56±0.05)%明显升高而后[d6(2.56±0.08)%、d8(2.64±0.14)%、d10(2.37±0.24)%]降低(P<0.05).iNKT 1细胞频率低于对照组且呈降低的趋势(P<0.05).iNKT 2细胞频率高于对照组在第4天肺部iNKT2细胞频率达到峰值,随后降低(P<0.05).iNKT 17细胞频率略高于对照组,差异无统计学意义(P>0.05),与第2天相比,第4、6、8、10天iNKT17细胞频率均显著升高(P<0.05).组间比较发现,干预组与对照组外周血血清IFN-γ、IL-4、IL-17A浓度比较,差异有统计学意义,α-GalCer干预组在不同时间点IFN-γ、IL-4、IL-17A 浓度比较,差异有统计学意义,与第2天相比,IFN-γ、IL-4水平于第6天、第8天均降低(P<0.05);IL-17A 水平于第6天降低(P<0.05).α-GalCer干预组的IFN-γ水平与对照组相比均有明显升高,差异有统计学意义(P<0.05);除第8天外,α-GalCer干预组的IL-4水平与对照组比较,差异有统计学意义(P<0.05);第2天、第8天α-GalCer干预组的IL-17A水平均低于对照组,差异有统计学意义(P<0.05).结论 腹腔注射α-GalCer对小鼠肺组织 iNKT细胞和亚群有不同的激活效应;iNKT细胞激活后,会由脉管系统迁入间质;腹腔注射α-GalCer可诱导血清细胞因子的变化,提示 α-GalCer注射可影响机体免疫功能.

目的 分析药物对罗氏Accu-Chek Performa血糖仪检测准确性的干扰.方法 采取4名健康人的肝素抗凝全血(采集3管/人,3 ml/管),分别对应A(对乙酰氨基酚组)、B(多巴胺组)、C(羟苯磺酸钙组)、D(甘露醇组)4组.所有的全血室温静置24 h后测量基础血糖值,再根据所需血糖浓度水平加入不同剂量50％的葡萄糖溶液,调整血糖浓度至低、中、高三个水平范围内(1.1～3.3 mmol/L、11.1～16.7 mmol/L和18.1～22.2 mmol/L).每个浓度水平分为3管(1 ml/管),标记为1号(不加药物)、2号(加入正常剂量的药物)、3号(加入有药物毒性剂量的药物).用罗氏血糖仪检测不同药物剂量各10次后收集数据进行计算整理.结果 对这4种不同药物在低、中、高浓度的血糖中进行测定,发现多巴胺对低浓度血糖浓度检测有影响,使检测结果偏高,而羟苯磺酸钙对不同浓度的血糖检测均有影响,使血糖浓度偏高,另外2种药物影响不明显.结论 多巴胺对低浓度的血糖,羟本磺酸钙对高中低浓度的血糖检测有干扰作用,在使用过程中应注意避免药物影响导致血糖检测值不准确.

Herein, the migration distribution and safety of specific phenotypic and functionally identified spleen-derived invariant natural killer T2 (iNKT2) cells after adoptive infusion in mice were studied. The proliferation and differentiation of iNKT cells were induced by intraperitoneal injection of α-galactosylceramide (α-GalCer) in vivo. Mouse spleens were isolated in a sterile environment. iNKT cells were isolated by magnetic-activated cell sorting columns (MS columns). Cytometric bead array (CBA) assay was used to detect cytokine secretion in the supernatant stimulated by iNKT cells. The basic life status of the mice was observed, and systematic quantitative scoring was conducted after injecting spleen-derived iNKT cells through the tail vein. An in vivo imaging system was used to trace the migration and distribution of iNKT cells in DBA mice. The percentage of the iNKT2 subgroup was the highest in 3 days after intraperitoneal injection of α-GalCer, and iNKT2 subsets accounted for more than 92% after separation and purification by magnetic-activated cell sorting (MACS). Anti-inflammatory cytokine IL-4 was mainly found in the supernatant of cell cultures. The adoptive infusion of iNKT cells into healthy mice resulted in no significant change in the basic life status of mice compared with the noninjected group. iNKT cells were detected in the lung, spleen, and liver, but no fluorescence was detected in lymph nodes and thymus. After dissecting the mice, it was found that there were no significant abnormalities in the relevant immune organs, brain, heart, kidney, lung, and other organs. Intraperitoneal injection of α-GalCer results in a large number of iNKT2 cells, mainly secreting anti-inflammatory cytokine IL-4, from the spleen of mice. After adoptive infusion, the iNKT2 cells mainly settled in the liver and spleen of mice with a satisfactory safety profile.

Non-small cell lung cancer (NSCLC) is a type of lung cancer which has a high mortality and low survival rate. Previous studies have revealed that long non-coding RNAs participate in tumorigenesis and metastasis in NSCLC. In the present study, the function of small nucleolar RNA host gene 12 (SNHG12) was investigated in NSCLC. Using reverse transcription-quantitative polymerase chain reaction analysis, it was identified that SNHG12 was significantly overexpressed in NSCLC specimens. Furthermore, overexpression of SNHG12 was identified to be associated with tumor progression and poor overall survival rates. Knockdown of SNHG12 in NSCLC cells could effectively induce cell apoptosis and suppress cell viability, proliferation, migration and invasion via inhibition of the epithelial-mesenchymal transition process. Furthermore, a direct interaction between microRNA (miR)-218 and the binding site of SNHG12 was identified. SNHG12 acted as an endogenous sponge for miR-218. Knockdown of SNHG12 upregulated the expression level of miR-218 as well as downregulating the Slug/zinc finger E-box-binding homeobox 2 EMT signaling pathway, and thus inhibited cell migration and invasion. Therefore, SNHG12 may serve as a key biomarker and a potential therapeutic target for the treatment of NSCLC.

目的 为准确测量脑脊液(CSF)样本中的乳酸脱氢酶(LDH),对血清LDH测量程序进行改良,并对正确度、精密度、线性范围等指标进行性能评估.方法 根据血清LDH测量程序,通过增加样本量的方法改善其分析性能,评估其正确度、精密度、线性范围.结果 当样本中LDH＜ 25 U/L时,改良测量程序的重复性、实验室内精密度均优于未改良测量程序,检测经参考方法定值稀释后的样本偏移分别为-4.4％、-6.57％,正确度优于未改良程序;改良测量程序线性方程为)y=0.995 6x-0.031 1,r=0.999 8,未改良程序线性方程为y=0.807 9x +0.817 8,r=0.989 5.结论 改良脑脊液LDH测量程序的精密度、正确度、线性均要优于未改良程序,为准确测量脑脊液中的LDH提供了一种简单、有效的方法.