Objective Veratraldehyde is an important pharmaceutical intermediate and chemical raw material. Taking common synthetic methods of Veratraldehyde into consideration, the study is aimed to further investigate a more efficient synthetic route in order to provide a solid foundation for the application of Veratraldehyde.Methods This article introduces an alternative method based on the Triton－B catalyzed olefin isomerization of methyl Eugenol followed by ozonation/reduction reaction.Results In the experiment,the optimal conditions was selected by investigating the influence of reaction time, reaction temperature and catalyst dosage on the yield of Veratraldehyde. The total yield of this 2－step reactions was 64% . Conclusion The above－mentioned synthesis method only requires raw materials that are easily acquired and its relative reaction conditions are mild, thus it will provide values in the industrial applications.

A sample preparation method was developed to simulate the process of intracellular metabolites metabonomics analysis of Escherichia coli. The Escherichia coli cell was firstly quenched with cold sodium chloride solution ( 0. 85 %，precooled at -80℃ for 15 min).The quenched bacterial cell was treated by using the technique of vacuum freeze-drying and liquid nitrogen freezing combined with ultrasonic processing to increase cell membrane penetrability. Finally，a cold aqueous solution of methanol (MeOH:H2O, 1:1，V/ V, 4 ℃) was used as extraction solvent to extract metabolites. In the present research，flow cytometry and OD value recovery were performed to evaluate the degree of cell damage caused by quenching at single cell level and at integral level respectively. The tested results indicated that the degree of damage to cells caused by cold sodium chloride solution was less than 5%. The peak quantity and the total ion intensity detected by LC-TOF in low collision energy were used to evaluate extraction effects. Three different cell membrane penetrability modes and 4 kinds of extraction solvents were investigated and compared.The results showed that the technique of liquid nitrogen freezing combined with ultrasonic processing for cell membrane penetrability and a cold aqueous solution of methanol (MeOH/H2O，1:1，V/V, 4℃) for extraction of metabolites had the best extraction effect(peak quantity was greater than or equal to 105，and total ion intensity was in the range of 106-107).Therefore，in this work，the freeze drying，grinding with liquid nitrogen and ultrasonic extraction were combined to extract metabolites. In this way，it effectively promoted cell lysis and improved the efficiency of extraction. The result of synthetic analysis showed that the method proposed here could meet the requirements of the metabonomics analysis of Escherachaa coli.

[摘 要] 多发性骨髓瘤是骨髓中浆细胞异常增生导致的恶性肿瘤，是第二大常见的血液系统恶性肿瘤。日益增多的生物治疗方法为多发性骨髓瘤治疗提供新的思路和方向，CAR-T细胞疗法更是为复发/难治性多发性骨髓瘤患者带来治愈新希望。已有多种靶向多发性骨髓瘤特异性靶标分子CAR-T细胞在临床试验中显示出较好的疗效，然而CAR-T细胞疗法仍存在疗效持续时间不够长、肿瘤易复发等问题，这可能与CAR-T细胞持续性不足、肿瘤细胞表面抗原表达丢失、抗原逃逸、免疫抑制微环境损害T细胞活性等因素相关。已有临床研究通过优化CAR设计、调整制备过程以产生富含特定T细胞亚群的CAR-T细胞、构建健康志愿者来源的通用型CAR-T细胞、引入修饰基因以调节免疫抑制微环境或改善CAR-T细胞增殖能力等方法来提高CAR-T细胞的效应功能并延长其持续作用时间，通过降低CAR结构中抗体免疫原性、引入开关机制等方法来提高CAR-T细胞疗法安全性。众多研究为多发性骨髓瘤的CAR-T细胞治疗注入新的活力，也为抗肿瘤免疫治疗提供新的方法与选择。

[摘 要] 目的：评价DC联合CIK治疗156例局部晚期或晚期胰腺癌的临床疗效。方法：回顾性分析2011年11月至2023年12月在东部战区总医院肿瘤科进行自体DC联合CIK治疗的156例局部晚期或晚期胰腺癌患者的临床资料。统计患者治疗前后血清肿瘤标志物、淋巴细胞亚群、细胞因子水平的变化、不良反应发生情况以及近期疗效、远期疗效。结果：156例胰腺癌患者中有92例治疗前后均进行了影像学检查，结果显示CR 0例，PR 0例，SD 42例，PD 50例，ORR为0%，DCR为45.65%。外周血CA199水平在治疗前后无显著差异，但有19例患者治疗后下降超过20%。治疗前后患者外周血CD3+、CD4+、CD8+、CD56+、CD25+淋巴细胞亚群水平和CD4+/CD8+ T细胞比值无统计学差异（P>0.05），治疗后患者外周血IL-2、IFN-γ的平均水平均显著高于治疗前（P<0.05），TNF-α和IL-6无显著差异（P>0.05）。156例患者mOS为8.53个月，1年累积生存率为39%，2年累积生存率为15%，3年累积生存率为15%，没有随访到5年的生存数据。治疗过程中未发生严重不良反应。结论：DC-CIK能使局部晚期和晚期胰腺癌患者产生抗肿瘤免疫反应，取得一定的客观疗效并可能使患者生存期延长。

[摘 要] 目的：采用生物信息学方法分析E3泛素连接酶（HECW2）在胃腺癌组织中的表达及其临床意义，为寻找胃腺癌诊断和预后生物标志物提供新的线索。方法：用R语言分析HECW2在泛癌中的表达及其与泛癌预后的关系。借助UCSC Xena、HPA、Kaplan-Meier Plotter等数据库分析HECW2在胃腺癌组织中的表达及其与临床病理特征之间的关系。采用WB法检测中国人胃腺癌组织及其癌旁组织中HECW2蛋白水平以验证数据库中的分析结果。借助TIMER和Cibersort数据库分析HECW2与胃腺癌免疫浸润的关系。通过LinkedOmics数据库对胃腺癌中HECW2进行GO功能分析和KEGG信号通路富集分析及相关性基因分析。结果：生物信息学分析结果表明，33种不同类型肿瘤中，包括胃腺癌在内的12种肿瘤中HECW2呈显著高表达（均P<0.05），WB法结果显示，中国人胃腺癌组织中HECW2也呈显著高表达（P<0.05）。HECW2表达水平越高，胃腺癌患者OS越短，包括CD4+ T细胞、巨噬细胞、中性粒细胞和树突状细胞在内的免疫细胞浸润丰度越高（均P˂0.01）。此外，HECW2相关信号通路主要富集于细胞外基质受体相互作用、黏着斑、细胞黏附和氧化磷酸化等生理病理过程（均P˂0.01）。结论：HECW2在胃腺癌组织中呈显著高表达且其与胃腺癌预后不良和免疫细胞浸润密切相关，有成为胃腺癌预后标志物和治疗靶点的潜力。

[Abstract] Objective: To investigate the effect of polyadenosine diphosphate ribose polymerase 1 (PARP1) on the proliferation and 5-FU resistance of gastric cancer cells and its potential mechanism. Methods: The tumor tissues and corresponding paracancerous tissues of 72 patients with gastric cancer who were treated in Central Hospital of Enshi Tujia and Miao Autonomous Prefecture from May 2018 to December 2019 were collected. AGS cells were transfected with siFUT8 to knock down FUT8 gene expression. qPCR and immunohistochemistry were used to detect the expression of PARP1 in gastric cancer and adjacent tissues. CCK-8, flow cytometry, and colony formation assay were employed to detect the effects of AG14361 on the proliferation, apoptosis and colony formation of AGS cells. MTT assay was used to detect the effect of AG14361 on the 5-FU sensitivity of gastric cancer cells. The overall distribution of differential genes in AGS cells treated with AG14361 was analyzed by mRNA sequencing, and related signaling pathways were analyzed by KEGG enrichment. qPCR and WB were used to detect the expression of α-1,6-fucosyltransferase (FUT8) in AGS cells and the interference effect of FUT8. CCK-8, flow cytometry, and colony formation assay were employed to detect the effects of AG14361 on the proliferation, apoptosis, and colony formation of AGS cells disturbed by siFUT8. Results: Compared with paracancer tissues, PARP1 expression was significantly increased in gastric cancer tissues (P<0.001). AG14361 can significantly inhibit the proliferation and colony formation of AGS cells, thus promoting the apoptosis of AGS cells (all P<0.01). AG14361 treatment reduced the IC50 of 5-FU against gastric cancer cells, especially against AGS cells, with IC50 decreased by more than 60%. mRNA sequencing results showed that FUT8 was a key glycosyltransferase of AG14361 in inhibiting the proliferation of AGS cells (P<0.05). Compared with the siNC group, treatment of AG14361 with IC50 significantly reversed the promotion of AGS cells proliferation caused by inerference with FUT8, promoted apoptosis and BAX protein expression, decreased Bcl2 protein expression and inhibited the increase in AGS cell colony formation caused by interference with FUT8 (all P<0.01). Conclusion: PARP1 can promote malignant transformation of gastric cancer cells by regulating N-glycosyltransferase FUT8. AG14361 can enhance the chemotherapy sensitivity of 5-FU, and PARP1 may become a potential target for gastric cancer treatment.

[摘 要] 目的：评价肿瘤特异性个体化多靶点树突状细胞-细胞因子诱导的杀伤细胞（DC-CIK）治疗晚期非小细胞肺癌（NSCLC）患者的临床疗效和安全性。方法：回顾性分析2019年10月1日至2022年10月31日东部战区总医院生物治疗科行肿瘤特异性个体化多靶点DC-CIK治疗晚期NSCLC患者的临床资料。统计NSCLC患者的临床疗效和不良反应，分析治疗前后血清中肿瘤标志物的变化，FCM检测患者治疗前后的淋巴细胞亚群和各种细胞因子的表达情况，用质谱仪检测治疗前后靶点的变化。结果: 共入组52例晚期NSCLC患者，其中女性21例、男性31例；年龄32~71岁，平均年龄（50.97±10.72）岁，中位年龄47.5岁。经DC-CIK治疗后，CR 0例，PR 0例，SD 27例，PD 25例。与治疗前比较，DC-CIK治疗后：（1）CEA和CYFRA21-1水平无显著改变，CA125水平显著低于治疗前（P<0.01）；（2）治疗后患者淋巴细胞亚群无显著变化；（3）治疗后患者外周血IL-2、IL-4、IFN-γ和TNF-α水平显著升高（均P<0.01），IL-6、IL-10及IL-17水平无明显变化；（4）治疗后靶点数下降明显。DC-CIK治疗过程中无严重不良反应发生。结论: 晚期NSCLC患者行肿瘤特异性个体化多靶点自体DC-CIK治疗是安全的，能使患者产生抗肿瘤免疫反应并得到一定的临床获益。

目的：通过向C57Bl/6J小鼠腹腔注射IFN-γ腺病毒（Ad-mIFN-γ）建立细胞因子释放综合征（CRS）的动物模型。方法：构建Ad-mIFN-γ及对照Ad-lacZ腺病毒载体，分别以MOI=100体外转染小鼠腹腔巨噬细胞，流式细胞术检测其对细胞mIFN-γ分泌的影响。将40只雌性C57Bl/6J小鼠按随机数字表法分为对照组、载体对照组、病毒低、中、高剂量组（每组8只），分别向各组小鼠腹腔注射PBS（200 μL）、Ad-lacZ（2×107 PFU/只）、Ad-mIFN-γ（5×106 PFU/只）、Ad-mIFN-γ（1.5×107 PFU/只）和Ad-mIFN-γ（2×107 PFU /只）。每日观测小鼠的体质量及生存情况；第3天时采用流式细胞术检测小鼠外周血和脾内单核细胞（CD11b+）、巨噬细胞（CD11b+/CD86+）比例，免疫荧光染色法检测脾内CD11b+的单核细胞比例；第9天时采用流式细胞术检测小鼠血清中细胞因子的分泌水平；第14天，采用颈椎脱臼法处死小鼠，H-E染色法观察小鼠肝、脾、肺和肾的病理和组织学变化。结果： Ad-mIFN-γ体外感染小鼠腹腔巨噬细胞，在第3天检测到巨噬细胞分泌mIFN-γ达到峰值（118.34±2.90）pg/mL，并在一周内持续高分泌mIFN-γ，Ad-lacZ对照组IFN-γ分泌水平较低后，第3天时为（0.17±0.08）pg/mL。小鼠腹腔注射Ad-mIFN-γ后，在14 d内病毒低、中剂量组无小鼠死亡，病毒高剂量组小鼠体质量持续减轻（P<0.001）；第3天，病毒高剂量组小鼠外周血和脾组织内单核细胞、巨噬细胞比例较对照组和中剂量组均显著增加（P<0.05或P<0.01）；第9天，病毒低、中、高剂量组小鼠血清中mIFN-γ、IL-6、单核细胞趋化蛋白-1（MCP-1）、IL-1、TNF-α等细胞因子的水平均显著升高（P<0.001）；10 d内病毒高剂量组小鼠死亡率达100%。组织病理检测可见病毒高剂量组小鼠的肝、脾、肺、肾组织有明显损伤。结论： Ad-mIFN-γ体外感染小鼠原代腹腔巨噬细胞后，可以快速分泌mIFN-γ；腹腔注射高剂量（2×107 PFU/只）Ad-mIFN-γ导致小鼠出现CRS典型表现，可作为CAR-T细胞治疗诱发CRS的动物模型。