[摘 要] 目的：探讨KH型剪切调节蛋白（KHSRP）靶向调控JAK1/STAT3信号轴对食管胃结合部腺癌（AEG）细胞增殖、迁移和侵袭及移植瘤生长与肺转移的影响。方法：收集2017年1月至2018年12月间在淮河医院确诊的64例AEG组织和癌旁组织标本及临床资料，采用免疫组化法观察AEG组织和癌旁组织中KHSRP的表达水平。qPCR法检测AEG细胞OE-19、TE-7、BIC-1、FLO-1、SK-GT-4、BE-3与正常食管上皮细胞Het-1A中KHSRP的表达差异。通过慢病毒载体对KHSRP进行敲减和过表达处理，分别转染OE-19与TE-7细胞、FLO-1与SK-GT-4细胞，实验分为sh-NC组、sh-KHSRP组和Vector组、KHSRP过表达组（KHSRP组）。采用qPCR法检测敲减或过表达效率，CCK-8法、Transwell小室法分别检测敲减或过表达KHSRP对AEG细胞增殖、迁移和侵袭的影响。构建小鼠异种移植瘤和肺转移模型，观察KHSRP对移植瘤体内生长和转移的作用。WB法验证KHSRP靶向JAK/STAT信号通路。细胞拯救实验验证KHSRP是否通过调节JAK1/STAT3信号通路促进AEG细胞的恶性进程。结果：与癌旁组织相比，AEG组织中KHSRP表达水平显著增高（P < 0.05或P < 0.01）。细胞功能实验分析显示，KHSRP过表达在体外均显著促进AEG细胞增殖、迁移和侵袭（P < 0.05或P < 0.01）。动物实验结果显示，KHSRP在裸鼠体内具有促进AEG细胞移植瘤生长与肺转移的作用（P < 0.05或P < 0.01）。在敲减KHSRP后，JAK/STAT信号通路中JAK1、STAT3磷酸化水平均明显降低，过表达KHSRP后情况则均反之（P < 0.05或P < 0.01）。细胞拯救实验显示，KHSRP可以逆转敲减JAK1/STAT3对细胞增殖、迁移和侵袭的抑制作用（P < 0.05或P < 0.01）。结论：KHSRP通过激活JAK1/STAT3信号通路调控AEG细胞转移的恶性进程，KHSRP有望成为AEG临床治疗的潜在靶点。

[摘 要] 目的：探究蛋白酪氨酸磷酸酶D（PTPRD）去甲基化通过PI3K/Akt/mTOR通路对胃癌细胞增殖、迁移及化疗耐药性的影响。方法：体外培养胃癌细胞MKN-74、MKN-45和人胃黏膜上皮细胞GES-1并检测PTPRD表达。常规培养MKN-45细胞及耐药MKN-45/5-FU细胞，分别转染PTPRD空载体（NC组、NC/5-FU组）、PTPRD过表达腺病毒（PTPRD组、PTPRD/5-FU组）、shRNA空载体（sh-NC组、sh-NC/5-FU组）、shRNA-PTPRD慢病毒（sh-PTPRD组、sh-PTPRD/5-FU组）和PTPRD过表达腺病毒 + 10 μmol/L 740Y-P处理（PTPRD + 740Y-P组、PTPRD + 740Y-P/5-FU组）。MTT法、划痕愈合实验检测各组细胞的增殖活力和迁移能力，细胞自噬实验检测细胞的自噬水平，WB法检测细胞中EMT和PI3K/Akt/mTOR通路相关蛋白的表达。采用0、2.5、5、10、20、40 μmol/L的5-aza处理MKN-45细胞，qPCR法、MTT法检测细胞中PTPRD mRNA表达和细胞增殖活力。结果：PTPRD mRNA和蛋白在胃癌细胞中均呈低表达（P < 0.05）。与MKN-45组相比，PTPRD组自噬体与自噬溶酶体数量、PTPRD、上皮钙黏素（E-cadherin）、BAX蛋白表达均增加（均P < 0.05），细胞增殖活力、细胞迁移率、p-PI3K、波形蛋白（vimentin）、p-Akt、p-mTOR蛋白表达均降低（均P < 0.05），sh-PTPRD组细胞增殖活力、细胞迁移率、p-PI3K、vimentin、p-Akt、p-mTOR蛋白表达均增加（均P < 0.05），自噬体与自噬溶酶体数量、PTPRD、E-cadherin、BAX蛋白表达均减少（均P < 0.05）；与PTPRD组相比，PTPRD + 740Y-P组细胞增殖活力、细胞迁移率、p-PI3K、vimentin、p-Akt、p-mTOR蛋白表达均增加（均P < 0.05），自噬体与自噬溶酶体数量、PTPRD、E-cadherin、BAX蛋白表达减少（均P < 0.05）。随着5-aza浓度的增加，MKN-45细胞中PTPRD mRNA表达增加、细胞增殖活力均降低（均P < 0.05）。与MKN-45/5-FU组相比，PTPRD/5-FU组细胞迁移率、细胞增殖活力均降低（均P < 0.05），sh-PTPRD/5-FU组细胞迁移率、细胞增殖活力均增加（均P < 0.05）；与PTPRD/5-FU组相比，PTPRD + 740Y-P/5-FU组细胞迁移率、细胞增殖活力均增加（均P < 0.05）。结论：PTPRD在胃癌细胞中呈低表达状态，PTPRD去甲基化可能通过抑制PI3K/Akt/mTOR信号通路抑制胃癌细胞增殖、迁移并增强其对化疗的敏感性。

Background/Aims: Systemic Inflammation (SI) is considered a key mechanism in disease progression and development of complications in decompensated liver cirrhosis. SI is mainly driven by portal hypertension and bacterial translocation. Transjugular intrahepatic portosystemic shunt (TIPS) insertion represents an effective treatment for portal hypertension. This study aims to investigate the impact of TIPS insertion on SI and bacterial translocation. Methods: We prospectively included 59 cirrhotic patients undergoing TIPS insertion. Blood samples were collected at TIPS insertion and follow-up (FU) 1, 3, 6, and 12 months thereafter. At all time points, we performed a comprehensive analysis of SI including 43 soluble inflammatory markers (SIMs), and surrogates of bacterial translocation (sCD14, sCD163). To investigate long-term kinetics of SI, C-reactive protein (CRP) and white blood cells (WBC) were retrospectively analyzed in a cohort of 177 patients up to 3 years after TIPS insertion. Results: At TIPS insertion, 30/43 SIMs, sCD14, and sCD163 measured significantly higher in cirrhotic patients compared to healthy controls. By FU6 25 SIMs and sCD14 measured at significantly lower levels compared to baseline. Interestingly, in patients with TIPS indication of refractory ascites, IL-6 decreased to levels documented in earlier stages of cirrhosis. In long-term follow-up, CRP levels significantly decreased after TIPS insertion, which translated into lower mortality in Cox regression analysis (HR 0.968, p=0.042). Notably, patients with residual ascites post-TIPS showed significantly higher CRP and IL-6 levels across all follow-ups compared to patients with resolved ascites. Conclusions Decreasing portal hypertension via TIPS insertion leads to a significant attenuation of SI and bacterial translocation over time.

Background/Aims: Systemic Inflammation (SI) is considered a key mechanism in disease progression and development of complications in decompensated liver cirrhosis. SI is mainly driven by portal hypertension and bacterial translocation. Transjugular intrahepatic portosystemic shunt (TIPS) insertion represents an effective treatment for portal hypertension. This study aims to investigate the impact of TIPS insertion on SI and bacterial translocation. Methods: We prospectively included 59 cirrhotic patients undergoing TIPS insertion. Blood samples were collected at TIPS insertion and follow-up (FU) 1, 3, 6, and 12 months thereafter. At all time points, we performed a comprehensive analysis of SI including 43 soluble inflammatory markers (SIMs), and surrogates of bacterial translocation (sCD14, sCD163). To investigate long-term kinetics of SI, C-reactive protein (CRP) and white blood cells (WBC) were retrospectively analyzed in a cohort of 177 patients up to 3 years after TIPS insertion. Results: At TIPS insertion, 30/43 SIMs, sCD14, and sCD163 measured significantly higher in cirrhotic patients compared to healthy controls. By FU6 25 SIMs and sCD14 measured at significantly lower levels compared to baseline. Interestingly, in patients with TIPS indication of refractory ascites, IL-6 decreased to levels documented in earlier stages of cirrhosis. In long-term follow-up, CRP levels significantly decreased after TIPS insertion, which translated into lower mortality in Cox regression analysis (HR 0.968, p=0.042). Notably, patients with residual ascites post-TIPS showed significantly higher CRP and IL-6 levels across all follow-ups compared to patients with resolved ascites. Conclusions Decreasing portal hypertension via TIPS insertion leads to a significant attenuation of SI and bacterial translocation over time.

Background/Aims: Systemic Inflammation (SI) is considered a key mechanism in disease progression and development of complications in decompensated liver cirrhosis. SI is mainly driven by portal hypertension and bacterial translocation. Transjugular intrahepatic portosystemic shunt (TIPS) insertion represents an effective treatment for portal hypertension. This study aims to investigate the impact of TIPS insertion on SI and bacterial translocation. Methods: We prospectively included 59 cirrhotic patients undergoing TIPS insertion. Blood samples were collected at TIPS insertion and follow-up (FU) 1, 3, 6, and 12 months thereafter. At all time points, we performed a comprehensive analysis of SI including 43 soluble inflammatory markers (SIMs), and surrogates of bacterial translocation (sCD14, sCD163). To investigate long-term kinetics of SI, C-reactive protein (CRP) and white blood cells (WBC) were retrospectively analyzed in a cohort of 177 patients up to 3 years after TIPS insertion. Results: At TIPS insertion, 30/43 SIMs, sCD14, and sCD163 measured significantly higher in cirrhotic patients compared to healthy controls. By FU6 25 SIMs and sCD14 measured at significantly lower levels compared to baseline. Interestingly, in patients with TIPS indication of refractory ascites, IL-6 decreased to levels documented in earlier stages of cirrhosis. In long-term follow-up, CRP levels significantly decreased after TIPS insertion, which translated into lower mortality in Cox regression analysis (HR 0.968, p=0.042). Notably, patients with residual ascites post-TIPS showed significantly higher CRP and IL-6 levels across all follow-ups compared to patients with resolved ascites. Conclusions Decreasing portal hypertension via TIPS insertion leads to a significant attenuation of SI and bacterial translocation over time.

Thermal analysis technology has emerged as a pivotal tool for the identification and quality control of traditional Chinese medicine （TCM） owing to its advantages of high sensitivity and capability for simultaneous multi-parameter detection. The application progress on thermogravimetric analysis （TGA）, differential thermal analysis （DTA）, and differential scanning calorimetry （DSC） in four key areas: authenticity identification of herbal medicines, optimization of processing techniques, evaluation of extract thermal stability, and construction of quality evaluation systems were summarized. Thermal analysis technology enables rapid authentication of medicinal materials by establishing a thermal fingerprint. When integrated with hyphenated techniques （e.g., FTIR and GC-MS）, it facilitates in-depth analysis of compositional differences in complex matrices. In Future, the development of thermal analysis databases and multi-technology integration will be expected to further promote the standardization of TCM quality control.

[摘 要] 目的：基于生物信息学分析和体内、体外实验研究长链非编码RNA00511（LINC00511）敲减对非小细胞肺癌细胞增殖、凋亡、侵袭等恶性生物学行为的影响，并初步探究其作用机制。方法：通过基因表达谱交互分析（GEPIA）数据库分析LINC00511在非小细胞肺癌的表达水平，及其与患者肿瘤分期、生存期等临床特征、肿瘤细胞恶性生物学行为有关基因的相关性；利用shRNA慢病毒载体构建LINC00511敲减的H1299肺癌细胞株，克隆形成实验、划痕愈合实验和流式细胞术分别检测对H1299细胞增殖、迁移、细胞周期和凋亡能力的影响，qRT-PCR检测相关调控基因表达，WB法检测肿瘤相关蛋白的表达；构建裸鼠皮下移植瘤模型，取瘤组织进行免疫组织化学实验检测Ki67表达情况。结果：GEPIA数据库分析表明LINC00511在非小细胞肺癌组织中表达水平升高，且与该病的临床分期情况相关（P < 0.05），LINC00511与肺癌中CASP3、CCNB1、CDK4等多种基因表达均有相关性（P < 0.01）；LINC00511敲减可抑制细胞的克隆形成和迁移能力、促进肺癌细胞凋亡并影响细胞周期进展（P < 0.05，P < 0.01）；LINC00511敲减可下调肺癌细胞CCNB、CDK4、TGF-β1基因的表达（P < 0.01），对CCND1、VEGFA基因表达无明显影响，LINC00511敲减可抑制细胞内MMP9、CTNNB1表达，上调CASP3的表达（P < 0.05，P < 0.01）；裸鼠体内实验证实，LINC00511敲减可抑制移植瘤体组织内Ki67的表达（P < 0.01）。结论：LINC00511在非小细胞肺癌组织中呈高表达，与肺癌临床分期和多种基因表达具有相关性，LINC00511敲减可能通过影响相关基因、蛋白的表达，抑制肺癌H1299细胞的恶性生物学行为。

[摘 要] 目的：利用昆虫杆状病毒表达系统（昆虫BEVS）表达糖酵解酶α-烯醇化酶（ENO1）及其3种酶活性位点缺失突变的ENO1蛋白ENO1-M1、ENO1-M2和ENO1-M3，为后续宫颈癌的代谢治疗研究奠定基础。方法：利用分子克隆技术将优化后ENO1序列插入pFastBacTM1载体，获得含有目的基因的重组质粒pFastBac-ENO1。分别缺失ENO1发挥糖酵解酶功能的3个活性位点，进行优化后将其插入pFastBacTM1载体，获得3个活性位点缺失的重组质粒pFastBac-M1、pFastBac-M2和pFastBac-M3。通过转座、转染后获得重组杆状病毒rBV-ENO1、rBV-M1、rBV-M2和rBV-M3，利用WB法对目的蛋白的表达及特异性进行检测。结果：成功扩增重组杆粒rBacmid-ENO1、rBacmid-M1、rBacmid-M2和rBacmid-M3，获得大小约2 000 bp的基因片段，与预期大小相符。昆虫BEVS可表达ENO1蛋白及其3个酶活位点缺失的重组蛋白ENO1-M1、ENO1-M2和ENO1-M3，其分子量约为52 000，与预期相符。WB法鉴定这些蛋白能与特异性标签His-tag发生反应。结论：通过昆虫BEVS成功表达目的蛋白ENO1及其酶活性位点缺失蛋白ENO1-M1、ENO1-M2和ENO1-M3，这些蛋白具有反应原性，为后续测定这些蛋白与ENO1单抗亲和力创造了条件。

[摘 要] 目的：探讨高良姜素（Gal）是否通过调节cGAS/STING信号通路影响骨肉瘤MG63细胞增殖、迁移、侵袭和凋亡。方法：体外培养人骨肉瘤MG63细胞，分别使用0、5、15、25、50、100、200 μmol/L的Gal培养48 h后，CCK-8法检测Gal对细胞活力的影响。将MG63细胞分为对照组（未处理细胞）、Gal低浓度组（Gal-L组，50 μmol/L Gal处理）、Gal高浓度组（Gal-H组，100 μmol/L Gal处理）和Gal-H+STING抑制剂组（Gal-H+H-151组，100 μmol/L Gal+8 μmol/L H-151处理）。采用EdU染色法、划痕愈合实验、Transwell小室法、流式细胞术检测各组细胞增殖活力、迁移、侵袭和凋亡能力，WB法检测各组细胞中cGAS、STING蛋白的表达水平。结果：与对照组比较，Gal-L组、Gal-H组细胞增殖活力、迁移、侵袭能力均显著降低（均P<0.05），细胞凋亡率和cGAS、STING蛋白表达水平均显著升高（均P<0.05）；与Gal-H组比较，Gal-H+H-151组细胞增殖活力、迁移和侵袭能力均显著升高（均P<0.05），细胞凋亡率和cGAS、STING蛋白表达水平均显著降低（均P<0.05）。结论：Gal可能通过激活cGAS/STING信号通路抑制骨肉瘤MG63细胞增殖、迁移、侵袭并促进细胞凋亡。

[摘 要] 目的：探讨解偶联蛋白2（UCP2）抑制剂京尼平（GEN）对人下咽癌FaDu细胞增殖及线粒体功能的影响。方法：使用不同浓度的GEN作用于FaDu细胞24 h，实验分为GEN 0（对照）、50、100、200和400 μmol/L组。采用CCK-8法检测各组细胞增殖能力，DCFH-DA探针及JC-1染色联合流式细胞术检测GEN对FaDu细胞活性氧（ROS）含量及线粒体膜电位的影响，激光共聚焦显微镜观察GEN对FaDu细胞线粒体膜通透性转换孔的影响，可见分光光度法检测细胞中乳酸的含量，WB法检测细胞中UCP2蛋白的表达变化。结果：与对照组相比，GEN可显著抑制FaDu细胞的增殖活力（P<0.05或P<0.01）、细胞中UCP2蛋白的表达（P<0.05），降低线粒体膜电位（P<0.05或P<0.01）、乳酸含量（P<0.000 1），改变细胞线粒体膜孔道通透性，提高细胞中ROS水平（P<0.05或P<0.01）。结论：GEN通过调节细胞中UCP2的表达水平进而影响细胞的氧化还原能力及线粒体功能，从而发挥抑制人下咽癌FaDu细胞增殖并诱导细胞凋亡的作用。