[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。

To investigate effects of verapamil on primary cultured human urethral scar fibroblasts (USFs) and to provide basis for protecting the formation of urethra scar.
 Methods: The cell proliferation was evaluated with the cell counting kit (CCK)-8 method after USFs were incubated various verapamil concentrations (50, 100, 150, 200, or 250 μmol/L) or solvent for 12, 24, or 48 h. The protein level of matrix metalloproteinase (MMP) was evaluated with ELISA after cells were incubated with verapamil (100 μmol/L) or solvent (control cells) for 24 h.
 Results: The proliferation of USFs was obviously suppressed after verapamil treatment, which was in a dose-dependent and time-dependent manner. Meanwhile, the protein levels of MMP-2 and MMP-9 in the verapamil treatment group increased obviously compared with those of the control groups (P<0.05).
 Conclusion: Calcium channel blockers may prevent the excessive formation of urethra scar by inhibiting the proliferation of urethral scar fibroblasts and enhancing the activity of MMP.
Calcium Channel Blockers ; pharmacology ; Cell Proliferation ; drug effects ; Cells, Cultured ; Cicatrix ; prevention & control ; Fibroblasts ; drug effects ; Humans ; Matrix Metalloproteinase 2 ; drug effects ; Matrix Metalloproteinase 9 ; drug effects ; Matrix Metalloproteinase Inhibitors ; pharmacology ; Up-Regulation ; drug effects ; Urethra ; cytology ; pathology ; Verapamil ; pharmacology

Objective To evaluate the efficacy of percutaneous pneumatic nephrolithotripsy combined with ultrasonic lithotripsy for the treatment of renal calculi. Methods From January 2005 to December 2006, 132 cases of renal calculi were treated by percutaneous pneumatic nephrolithotripsy and ultrasonic lithotripsy. Percutaneous nephrolithotripsy was performed under the guidance of B-ultrasonography, and then the calculi were removed by pneumatic nephrolithotripsy combined with ultrasonic lithotripsy.Results Single-tract procedure was performed on 122 cases, while two-tract in the other 10. One-stage lithotripsy was performed on 127 cases, and two-stage operation was carried out in 5. The operation time ranged from 30 to 150 minutes with a mean of (60?12) minutes. The stone clearance rate was 90.2% (119/132) one week after the operation. Five patients had 200-to 500-ml hemorrhage 1 to 4 days after the operation, and were cured by blood transfusion and anti-infection treatments. No serious complications were found in the other patients. The nephrostomy tube was withdrawn 4 to 7 days (mean 6 days) after the operation. The hospital stay was 6 to 22 days with a mean of 15 days. Among the patients, 98 were followed up for 3 to 20 months (mean 13 months). No hemorrhage, infection, or recurrence of renal calculi was found during this period. Conclusion Percutaneous pneumatic nephrolithotripsy combined with ultrasonic lithotripsy is safe, effective, with mild surgical trauma and a few complications for patients with renal calculi.