[摘 要] 目的：探讨Ⅱ型固有淋巴细胞（ILC2）-双调蛋白（AREG）-调节性T（Treg）细胞轴在宫颈癌免疫微环境调控中的作用及机制。方法：收集2021年5月—2022年5月于新疆医科大学第一附属医院收治的Ⅰ~ⅡA期宫颈癌患者肿瘤组织（n = 8），以子宫肌瘤手术患者的正常宫颈组织（n = 8）作为对照；另收集全分期宫颈癌患者的外周血样本（n = 30），并以健康志愿者外周血作为对照（n = 30）。利用GEPIA数据库分析AREG与叉头框蛋白P3（Foxp3）的mRNA表达水平。采用多重免疫荧光技术、流式细胞术检测组织及外周血中ILC2、Treg细胞浸润水平；通过ELISA、IHC和WB法验证AREG、Foxp3及IL-10的表达水平，并对ILC2、AREG、Treg细胞及IL-10进行相关性分析。体外分离宫颈癌患者ILC2与PBMC，分别使用重组人IL-33（rhIL-33）、抗人IL-33抗体（α-IL-33）及重组人AREG（rhAREG）、抗人AREG抗体（α-AREG）进行干预；采用CCK-8法与流式细胞术检测不同浓度rhAREG对HeLa、SiHa细胞增殖及凋亡的影响，ELISA检测上清中AREG、IL-10浓度，流式细胞术检测Treg细胞比例变化。此外，本研究还比较了宫颈癌患者手术前后外周血中ILC2、AREG、Treg细胞及IL-10的水平差异。结果：宫颈癌患者组织及外周血中ILC2浸润水平和AREG表达显著高于对照组，Treg细胞比例、Foxp3及IL-10表达亦明显上调（P < 0.05）；相关性分析显示，ILC2、Treg细胞、AREG与IL-10彼此正向关联。体外实验表明，不同浓度rhAREG对宫颈癌细胞（HeLa、SiHa）的增殖及凋亡无明显作用（P > 0.05）；rhIL-33可激活ILC2并上调AREG分泌（P < 0.01），而α-IL-33可逆转该效应（P < 0.05）；rhAREG可促进Treg细胞分化及IL-10分泌（P < 0.001），α-AREG则显著逆转上述作用（P < 0.01）。此外，宫颈癌患者术后外周血中ILC2、Treg细胞、AREG及IL-10水平均显著降低（P < 0.000 1）。结论：ILC2-AREG-Treg免疫调控轴异常激活，可能通过介导免疫抑制性肿瘤微环境形成参与宫颈癌进展。

Objective To investigate the effect and underlying mechanism of miR-137 on athero-sclerosis(AS)plaques in apolipoprotein E(ApoE)gene knockout(ApoE)mice.Methods Sixty ApoE-/-mice were fed with high-fat diet for 12 weeks to establish an AS model.Then they were assigned into AS group,negative control group,miR-137 group,Ad negative control group,and combination group,with 12 mice in each group;Another 12 wild-type C57BL/6 mice fed with chow diet were subjected as the Control group.Fully automated biochemical analyzer was applied to detect serum lipid levels,including total cholesterol(TC),triglycerides(TG),low-density lipo-protein cholesterol(LDL-C),and high-density lipoprotein cholesterol(HDL-C).ELISA was con-ducted to detect the levels of serum inflammatory factors,including TNF-α,IL-4,IL-6,and IL-10.HE staining was used to observe the morphological changes of mouse aortic tissue.Oil red O stai-ning was employed to observe the overall formation of aortic plaques.Immunofluorescence stai-ning was utilized to detect the expression of inducible nitric oxide synthase(iNOS)and arginase-1(Arg-1)in the aortic tissue.Real-time qPCR was applied to detect the mRNA expression of miR-137 and sex determining region Y box protein 4(SOX4)in the aorta.Results The miR-137 group has significantly lower serum levels of TG,TC,LDL-C,TNF-α and IL-6,and higher levels of HDL-C,IL-10 and IL-4 when compared with the AS group and negative control group(P＜0.05).The combination treatment resulted in increased serum levels of TG,TC,LDL-C,TNF-αand IL-6,while decreased levels of HDL-C,IL-10 and IL-4 in comparison with the Ad negative control group(P＜0.05).Larger aortic plaque area,more severe overall aortic plaque injury and stronger iNOS fluorescence intensity were observed in the AS group than the control group(P＜0.05).Treatment of miR-137 reversed above histological changes,resulting in smaller aortic plaque area,attenuated overall aortic plaque injury,decreased iNOS fluorescence intensity,and elevated Arg-1 fluorescence intensity when compared with the AS group and negative control group(P＜0.05).Compared with the Ad negative control group,the aortic plaque area,overall aortic plaque injury and iNOS fluorescence intensity were increased,while the Arg-1 fluorescence intensity was significantly decreased in the combined group(P＜0.05).Double luciferase assay showed that the luciferase activity of SOX4-containing wild-type cells was significantly decreased after transfection of miR-137 mimics when compared with transfection of mimics negative control(0.37±0.05 vs 1.00±0.08,P＜0.05).Conclusion Overexpression of miR-137 inhibits the activa-tion of rat sarcoma/mitogen-activated protein kinase pathway probably by down-regulating SOX4 expression,and then suppress M1 macrophage polarization and promote M2 macrophage polariza-tion,reduces inflammatory response and the formation of AS plaques.

Objective To investigate the effect and underlying mechanism of miR-137 on athero-sclerosis(AS)plaques in apolipoprotein E(ApoE)gene knockout(ApoE)mice.Methods Sixty ApoE-/-mice were fed with high-fat diet for 12 weeks to establish an AS model.Then they were assigned into AS group,negative control group,miR-137 group,Ad negative control group,and combination group,with 12 mice in each group;Another 12 wild-type C57BL/6 mice fed with chow diet were subjected as the Control group.Fully automated biochemical analyzer was applied to detect serum lipid levels,including total cholesterol(TC),triglycerides(TG),low-density lipo-protein cholesterol(LDL-C),and high-density lipoprotein cholesterol(HDL-C).ELISA was con-ducted to detect the levels of serum inflammatory factors,including TNF-α,IL-4,IL-6,and IL-10.HE staining was used to observe the morphological changes of mouse aortic tissue.Oil red O stai-ning was employed to observe the overall formation of aortic plaques.Immunofluorescence stai-ning was utilized to detect the expression of inducible nitric oxide synthase(iNOS)and arginase-1(Arg-1)in the aortic tissue.Real-time qPCR was applied to detect the mRNA expression of miR-137 and sex determining region Y box protein 4(SOX4)in the aorta.Results The miR-137 group has significantly lower serum levels of TG,TC,LDL-C,TNF-α and IL-6,and higher levels of HDL-C,IL-10 and IL-4 when compared with the AS group and negative control group(P＜0.05).The combination treatment resulted in increased serum levels of TG,TC,LDL-C,TNF-αand IL-6,while decreased levels of HDL-C,IL-10 and IL-4 in comparison with the Ad negative control group(P＜0.05).Larger aortic plaque area,more severe overall aortic plaque injury and stronger iNOS fluorescence intensity were observed in the AS group than the control group(P＜0.05).Treatment of miR-137 reversed above histological changes,resulting in smaller aortic plaque area,attenuated overall aortic plaque injury,decreased iNOS fluorescence intensity,and elevated Arg-1 fluorescence intensity when compared with the AS group and negative control group(P＜0.05).Compared with the Ad negative control group,the aortic plaque area,overall aortic plaque injury and iNOS fluorescence intensity were increased,while the Arg-1 fluorescence intensity was significantly decreased in the combined group(P＜0.05).Double luciferase assay showed that the luciferase activity of SOX4-containing wild-type cells was significantly decreased after transfection of miR-137 mimics when compared with transfection of mimics negative control(0.37±0.05 vs 1.00±0.08,P＜0.05).Conclusion Overexpression of miR-137 inhibits the activa-tion of rat sarcoma/mitogen-activated protein kinase pathway probably by down-regulating SOX4 expression,and then suppress M1 macrophage polarization and promote M2 macrophage polariza-tion,reduces inflammatory response and the formation of AS plaques.

OBJECTIVETo investigate the role of miR-144-3p in regulating osteogenic differentiation of bone marrow mesenchymal stem cells and predict its target genes.

METHODSRat bone marrow mesenchymal stem cells (BMSCs) with induced osteogenic differentiation were examined for the expressions of Runx2, OCN and miR-144-3p. The effects of transfection with a miR-144-3p mimic or a miR-144-3p inhibitor were tested on the osteogenic differentiation of the BMSCs. The changes in the expressions of the predicted target of miR-144-3p in the BMSCs during induced osteogenic differentiation were examined using Western blotting and qRT-PCR.

RESULTSRat BMSCs with induced differentiation into osteoblasts exhibited a progressive increase in the expressions of Runx2 and OCN (two markers of osteogenic differentiation), while the expression of miR-144-3p gradually decreased during the differentiation till reaching the lowest level at 21 days of induction. In rat BMSCs, transfection with the miR-144-3p mimic significantly decreased ALP activity ( < 0.05) wile transfection with the miR-144-3p inhibitor significantly increased ALP activity ( < 0.05) in rat BMSCs. Analysis based on miRanda, microRNA.org database and TargetScan suggested that Smad4 was the most likely target gene of miR-144-3p. The results of qRT-PCR showed no significant differences in expression levels of Smad4 among the cells with different treatments ( > 0.05), while Western blotting revealed a significantly decreased expression of Smad4 in the cells transfected with miR-144-3p mimics and an increased Smad4 expression in the cells transfected with the miR-144-3p inhibitor as compared with the control cells ( < 0.05).

CONCLUSIONSmiR-144-3p participates in the regulation of osteogenic differentiation of rat BMSCs, and its inhibitory effect on osteogenic differentiation is achieved probably by decreasing the expression level of Smad4.

Objective To understand the life attitude of college students in earthquake-stricken area. Methods 166 college students were conducted by life value scale (LVS) and suicide attitude questionnaire (QSA). Results ①On the LVS, males( (1.85 ±0.57) , (1.73±0.55) ) got significantly higher scores than females ((1.66±0.43),(1.54±0.40)) on survival values-oriented and spiritual values-oriented, and lower scores on tool values-oriented ((2.59±0.46 vs (2.76±0.51) ) , there was significant difference on overcome values-oriented between different grades(F = 3.710, P = 0.013). On the QSA, males (3.01 ±0.51) got significantly lower scores than females (3.28 ±0.51) on the cognition of suicidal behavior difference (t=3.110, P=0.002). There were significant differences on attitude to relationship of suicides between different gender and areas (P<0.05) , and attitude to euthanasia between different gender, grades and subjects (P<0.05). ②Social valuesoriented can predict cognition of suicidal behavior, personal and tool values-oriented can predict attitude to relationship of suicides, the total of life value can predict cognition of suicidal behavior and attitude to relationship of suicides. Conclusion The life value can predict the suicide attitude of college students in earthquake-stricken area, thus the life education should be enhanced to improve their quality of life.

Objective To evaluate the therapeutic effects of home made Nitrol stents in the teatment of malignant or benign tracheobronchial stenosis.Methods Thirteen patients with malignant or benign tracheobronchial stenosis were treated by Nitrol stents. The stenosed sites located in trachea in 5, tracheobronchi in 6, main bronchus in 2. All cases were malignant except one was benign. Results 15 stents were successfully placed the expected position with dyspnea rapidly improved. The average survival time was 11.4 months. Conclusions It is an effective way to place Nitrol stent in treatment of tracheobronchial stenosis under x ray guidance with fibertracheobronchoscopy.