丹参酮生物合成相关SmERF108转录因子的鉴定及其靶基因分析＊

目的 鉴定与丹参酮合成相关转录因子AP2/ERF家族中SmERF108转录因子，并分析转录因子SmERF108的靶基因。方法 利用生物信息学在线分析平台如NCBI、PFAM等分析SmERF108的序列特征；MEGA-X软件用于构建SmERF108与不同功能转录因子的系统进化树；拟南芥原生质体转化法鉴定SmERF108蛋白的亚细胞定位；通过实时荧光定量PCR（Real-time qPCR）对SmERF108基因在丹参不同器官和组织差异表达进行检测；利用酵母体系对SmERF108的转录激活活性进行探究并用酵母单杂技术确定其靶基因。结果 SmERF108具有典型的AP2/ERF保守结构域，属于ERF-B3亚组，系统进化树和保守基序分析显示SmERF108与丹参中SmERF128、青蒿中AaERF2亲缘关系较近且保守基序分布一致；亚细胞定位显示SmERF108蛋白定位于细胞核；SmERF108基因在周皮中表达最高，且呈现周皮（R1）>韧皮部（R2）>木质部（R3）的规律；酵母自激活验证SmERF108具有转录激活活性，同时酵母单杂交确认其能与关键酶基因SmCPS1启动子结合。结论 鉴定到丹参中一个新转录因子SmERF108，生物信息学分析及差异表达分析预测与丹参酮合成相关，分子互作初步证实靶基因为SmCPS1二萜环化酶。     

Effect of Yinqi Ointment on Wound Morphology and Growth Factor in Treating Diabetic Foot Ulcer

Objective: To investigate the effect of Yinqi ointment on wound morphology and growth factor in treating diabetic foot ulcer(DFU).Methods: From December 2016 to December 2017, 92 cases of DFU with deficiency of both Qi and Yin syndrome were randomly divided into treatment group and control group(44 cases in each group). The treatment group was treated with Yinqi ointment, while the control group was treated with mupirocin ointment. After 4 weeks of treatment, the ulcer healing effect, ulcer area, granulation tissue, epithelial tissue coverage,pain score, and dynamic analysis of vascular endothelial growth factor(VEGF), epidermis growth factor(EGF), and basic fibroblast growth factor(bFGF) in local granulation tissue were statistically analyzed before and after treatment in both groups. Results: The total effective rate was 88.37% in the treatment group and 74.42% in the control group. The wound reduction rate, epithelial tissue coverage rate, granulation tissue growth rate, and local pain relief rate in the treatment group were significantly superior to those in the control group(P 0.05). Through the local granulation detection, the treatment group and the control group have increased VEGF, EGF, and bFGF, but the treatment group increased the role of growth factor than the control group. Conclusion: Yinqi ointment can promote the healing of DFU, and its mechanism may be related to the increase of the content of growth factor in granulation tissue.     

麻黄细辛附子汤对肾阳虚外感小鼠及正常小鼠的效-毒作用研究

目的研究麻黄细辛附子汤(MXF)干预肾阳虚外感小鼠的药效作用及其对正常小鼠的毒性作用差异,并考察其效-毒作用机制。方法BABL/C雄性小鼠随机分为正常组、正常毒性低、中、高剂量组;肾阳虚外感模型组;肾阳虚外感低、中、高剂量组,连续灌胃给药6 d,记录体重、肛温等药理指标并观察小鼠的一般体征,检测生化指标、炎症因子。结果药效实验中,与正常组小鼠相比,肾阳虚外感模型组小鼠生化指标肌酸激酶(CK)、肌酸激酶-同工酶(CK-MB)、羟丁酸脱氢酶(α-HBDH)、乳酸盐脱氢酶(LDH)、丙氨酸氨基转移酶(ALT)、天冬氨酸转移酶(AST)、碱性磷酸酶(ALP)、尿素(urea)含量明显升高(P<0.05),炎症因子白介素-6(IL-6)、单核细胞去化蛋白(MCP-1)、肿瘤坏死因子-α(TNF-α)、白介素-10(IL-10)、干扰素-γ(INF-γ)以及白介素12(IL-12p70)含量升高明显(P<0.05)。经给药治疗后,各给药组CK、ALT、urea等生化指标含量较模型组有所回调,各给药组IL-6、IL-10、IFN-γ等炎症因子含量较模型组有不同程度的回调;毒性研究中,各给药组生化指标CK、urea、半胱氨酸(CYS-C)的含量较正常组显著升高(P<0.05),炎症因子白介素-10(IL-10)含量明显上升、白介素-12(IL-12p70)含量明显下降(P<0.05)。结论经MXF给药治疗后,肾阳虚外感给药组小鼠的一些生化指标均有不同程度的回调,MXF可通过调节IL-6、IL-10、IFN-γ、MCP-1、肿瘤坏死因子-α(TNF-α)等炎症因子改善机体的炎症反应;而正常小鼠给予不同剂量的MXF后心脏、肝脏和肾脏功能出现失调,机体通过细胞转移、打破免疫应答的平衡,从而使正常小鼠出现炎症反应。     

六味地黄汤干预抑郁大鼠学习记忆的作用机制

目的:探讨六味地黄汤对慢性抑郁大鼠记忆障碍的改善作用,并探讨其可能的作用机制。方法:Wistar雌性大鼠随机分为正常组(生理盐水)、慢性不可预见性温和应激(CUMS)模型组(生理盐水)、六味地黄汤低、中、高剂量组(2. 60,7. 81,23. 50 g·kg~(-1)·d~(-1))。除正常组外,其余各组造成CUMS模型。每周称体质量,观察其行为学指标变化;实时荧光定量聚合酶链式反应(Real-time PCR)检测其海马G蛋白偶联雌激素受体(GPR30),胞内磷脂酰肌醇激酶(PI3K),环磷酸腺苷反应元件结合蛋白(CREB),脑源性神经营养因子(BDNF) mRNA的表达;酶联免疫吸附测定(ELISA)检测血清中雌激素含量。结果:与正常组比较,模型组体质量、活动能力、兴趣等明显降低(P 0. 05,P 0. 01);与模型组比较,六味地黄汤2. 60,7. 81,23. 50 g·kg~(-1)可明显提高CUMS大鼠糖水偏好度(P 0. 01)和旷场实验的站立次数(P 0. 01);7. 81,23. 50 g·kg~(-1)明显提高旷场实验的总距离(P 0. 05,P 0. 01);2. 60,7. 81 g·kg~(-1)可缩短水迷宫实验寻台潜伏期(P 0. 01);7. 81 g·kg~(-1)可增加血清中雌激素含量(P 0. 05);CUMS模型组大鼠海马组织内的GPR30,PI3K,CREB,BDNF mRNA表达明显下降(P 0. 05,P 0. 01),六味地黄汤2. 60 g·kg~(-1)显著增加海马组织内的GPR30,CREB mRNA表达(P 0. 05,P 0. 01),7. 81 g·kg~(-1)明显增加海马组织内的GPR30,PI3K,CREB,BDNF mRNA表达量(P 0. 05,P 0. 01)。结论:六味地黄汤具有抗抑郁作用,逆转CUMS大鼠抑郁样行为及学习记忆障碍,其中中剂量组药效最显著。其作用机制可能与增加血清中雌激素和提高大鼠海马GPR30,PI3K,CREB,BDNF mRNA表达有关。     

Comparison of survival between adolescent and young adult vs older patients with hepatocellular carcinoma

BACKGROUNDDue to the special clinical features and biologic characteristics of adolescent and young adult (AYA) cancers, AYA cancers are different from cancers in children and elderly individuals. However, there are few reports on AYA hepatocellular carcinoma (HCC).AIMTo investigate the overall survival (OS) of AYA (15-39 years) and elderly (40-74 years) patients with HCC.METHODSThe data of all the HCC cases were extracted from the Surveillance, Epidemiology, and End Results database from 2004 to 2015 and were then divided into two groups based on age: AYA group (15-39 years) and older group (40-74 years). Kaplan-Meier curves and log-rank tests were used to compare the OS of the two groups. Propensity score matching (PSM) was employed to analyze the OS difference between the two groups. The Cox proportional hazards regression model was used to perform multivariate analysis to explore the risk factors for OS of HCC patients.RESULTSCompared to elderly cancer patients, AYA patients with HCC had a worse Surveillance, Epidemiology, and End Results stage, including the distant stage (22.1% vs 15.4%, P < 0.001), and a more advanced American Joint Committee on Cancer (AJCC) stage, including AJCC III and IV (49.2% vs 38.3%, P < 0.001), and were more likely to receive surgery (64.5% vs 47.5%, P < 0.001). Before PSM, the AYA group had a longer survival in months (median: 20.00, interquartile range [IQR]: 5.00-62.50) than the older group (median: 15.00, IQR: 4.00-40.00) (P < 0.001). After PSM, the AYA group still had a longer survival in months (median: 21.00, IQR: 5.00-64.50) than the older group (median: 18.00, IQR: 6.00-53.00) (P < 0.001). The Cox proportional hazards regression model showed that advanced age (hazard ratio [HR] = 1.405, 95%CI: 1.218-1.621, P < 0.001) was a risk factor for OS of HCC patients. In the subgroup analysis, the Cox proportional hazards regression model showed that in AJCC I/II HCC patients, advanced age (HR = 1.749, 95%CI: 1.352-2.263, P < 0.001) was a risk factor for OS, while it was not a risk factor in AJCC III/IV HCC patients (HR = 1.186, 95%CI: 0.997-1.410, P = 0.054) before PSM. After PSM, advanced age (HR = 1.891, 95%CI: 1.356-2.637, P < 0.001) was still a risk factor for OS in AJCC I/II HCC patients, but was not a risk factor for OS in AJCC III/IV HCC patients (HR = 1.192, 95%CI: 0.934-1.521, P = 0.157) after PSM.CONCLUSIONAYA patients with HCC have different clinical characteristics from older adults. In different AJCC stages, the two groups of patients have different OS: In AJCC I/II HCC patients, advanced age is a risk factor for OS, but it is not a risk factor for OS in the AJCC III/IV HCC patient group.     

First‐in‐human phase I study of E7090, a novel selective fibroblast growth factor receptor inhibitor,in patients with advanced solid tumors

Fibroblast growth factor receptors (FGFR) are a family of transmembrane receptor tyrosine kinases involved in regulating cellular processes. FGFR mutations are implicated in oncogenesis, representing therapeutic potential in the form of FGFR inhibitors. This phase I, first‐in‐human study in Japan evaluated safety and tolerability of E7090, a potent selective FGFR1‐3 inhibitor, in patients with advanced solid tumors. Dose escalation (daily oral dose of 1‐180 mg) was carried out to assess dose‐limiting toxicity (DLT), maximum tolerated dose, and pharmacokinetics. Pharmacodynamic markers (serum phosphate, fibroblast growth factor 23, and 1,25‐(OH)₂‐vitamin D) were also evaluated. A total of 24 patients refractory to standard therapy or for whom no appropriate treatment was available were enrolled. No DLT were observed up to the 140‐mg dose; one patient in the 180‐mg cohort experienced a DLT (increased aspartate aminotransferase/alanine aminotransferase, grade 3). The maximum tolerated dose was not reached. Dose‐dependent increases in the maximum concentration and area under the curve from time 0 to the last measurable concentration were observed up to 180 mg. Dose‐dependent increases were observed in all pharmacodynamic markers and plateaued at 100‐140 mg, indicating sufficient FGFR pathway inhibition at doses ≥100 mg. In conclusion, E7090 showed a manageable safety profile with no DLT at doses ≤140 mg. Maximum tolerated dose was not determined. The recommended dose for the follow‐up expansion part, restricted to patients with tumors harboring FGFR alterations, was determined as 140 mg, once daily.     

基于网络药理学的中期诊疗方干预新型冠状病毒肺炎(COVID-19)作用机制研究

目的通过网络药理学方法研究新型冠状病毒肺炎(COVID-19)中期诊疗方(XGZQZLF)治疗COVID-19的作用机制。方法采用网络药理学方法,以中药系统药理学数据库、GeneCards数据库等为支撑,通过Cytoscape构建"药物-活性成分-靶点-疾病"网络,进一步进行基因富集分析和通路注释,研究XGZQZLF治疗COVID-19的物质基础和作用机制。结果筛选出XGZQZLF有效成分80个,通过调控蛋白激酶(AKT1)、白细胞介素6(IL-6)、丝裂原活化蛋白激酶3(MAPK3)、血管内皮生长因子A(VEGFA)、半胱氨酸蛋白酶3(CASP3)等208个肺炎相关靶点,以及主要通过调控TNF信号通路、IL-17信号通路、Th17细胞分化、缺氧诱导因子1(HIF-1)信号通路等炎症和免疫相关通路发挥治疗COVID-19的作用。结论 XGZQZLF通过多成分、多靶点、多通路协同作用发挥治疗COVID-19的作用,为XGZQZLF治疗COVID-19的生物机制研究提供数据支撑,同时为COVID-19药物研发提供参考。     

靶向沉默髓鞘转录因子MyT1对脑胶质瘤细胞恶性生物学行为的影响

目的 探讨靶向沉默髓鞘转录因子1（MyT1）对人脑胶质瘤细胞迁移、侵袭和黏附的影响和相关分子机制。方法 设计特异性靶向沉默MyT1基因的shRNA，包装慢病毒后感染人脑胶质瘤U-118MG和U-87MG细胞，qPCR和Western blot检测两种细胞中MyT1的表达水平，BrdU实验、细胞划痕实验、Transwell实验和细胞黏附实验分别检测两种细胞的迁移、侵袭和黏附能力的变化，qPCR检测细胞黏附和肿瘤转移相关基因的表达水平。结果 在HEK293T细胞中包装了特异性靶向MyT1基因的shRNA慢病毒，并成功感染了U-118MG和U-87MG细胞；两种细胞中MyT1 mRNA和蛋白表达水平均显著下调（均P<0.05），细胞的迁移、侵袭和黏附能力均有一定程度的降低（均P<0.05），细胞黏附相关基因表达水平显著下降，肿瘤转移相关基因表达水平显著上升（均P<0.05）。结论 靶向沉默MyT1基因对人脑胶质瘤U-118MG和U-87MG细胞的迁移、侵袭和黏附能力有抑制作用，其机制可能与调控细胞黏附和肿瘤转移相关基因的表达有关。MyT1基因的表达，可能是脑胶质瘤诊疗的一个潜在靶标。     

肾炎方联合缬沙坦分散片对慢性肾小球肾炎患者的疗效与肾功能的影响

目的:探究肾炎方联合缬沙坦分散片治疗慢性肾小球肾炎患者的疗效及对肾功能的影响。方法:选取2016年2月至2019年2月重庆医科大学附属一医院合川医院肾脏内科收治的慢性肾小球肾炎患者122例作为研究对象,随机分为对照组和观察组,每组61例。对照组患者采用下缬沙坦分散片治疗,观察组在对照组基础上采用自拟肾炎方与缬沙坦分散片联合治疗,分析比较2组患者的肾功能指标的动态变化、炎性因子的表达水平、临床疗效及治疗后不良反应。结果:治疗后观察组TC、TG、BUN、Scr及24 hUPE5项指标的动态变化均低于对照组,差异有统计学意义(P<0.05),且观察组ALB动态变化高于对照组,差异有统计学意义(P<0.05);治疗后观察组IL-6、IL-10、CRP及TNF-α水平明显低于对照组,差异有统计学意义(P<0.05),观察组治疗有效率(98.4%)优于对照组(86.9%),差异有统计学意义(P<0.05);观察组不良反应发生率(6.6%)低于对照组(31.1%),差异有统计学意义(P<0.05)。结论:采用自拟肾炎方与缬沙坦分散片联合的治疗方案可增强疗效,提升肾功能,降低促炎性因子的表达水平及不良反应,值得在临床中推广普及。     

益气养阴解毒方联合环磷腺苷葡胺注射液治疗病毒性心肌炎35例

目的:探讨益气养阴解毒方联合环磷腺苷葡胺注射液对病毒性心肌炎患者心功能及外周血清肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、转化生长因子-β1(transforming growth factor-β1,TGF-β1)和干扰素-γ(interferon-γ,IFN-γ)水平的影响。方法:将70例病毒性心肌炎患者按照随机数字法分为对照组和观察组,每组各35例。两组患者均给予常规治疗,对照组在常规治疗的基础上给予环磷腺苷葡胺注射液静脉滴注,观察组在对照组的基础上联合益气养阴解毒方治疗。比较两组患者的临床疗效及治疗前后左室短轴缩短分数(left ventricular shortening fraction,LVFS)、左心室射血分数(left ventricular ejection fraction,LVEF)、心脏指数(cardiac index,CI)、TNF-α、TGF-β1和IFN-γ水平。结果:观察组有效率为88.57%,对照组有效率为68.57%,两组患者有效率比较,差异有统计学意义(P<0.05)。两组患者治疗后LVEF、LVFS高于本组治疗前,且观察组高于对照组,差异具有统计学意义(P<0.05)。两组患者治疗后TGF-β1、TNF-α水平低于治疗前比较,IFN-γ高于治疗前,且观察组TGF-β1、TNF-α低于对照组,IFN-γ高于对照组,差异均有统计学意义(P<0.05)。两组患者不良反应发生率比较,差异无统计学意义(P>0.05)。结论:益气养阴解毒方联合环磷腺苷葡胺注射液治疗病毒性心肌炎疗效更显著,可减轻机体的炎症反应,恢复患者心功能。