关键词： 人乳头瘤病毒   子宫颈癌  

摘要： 大量研究证实,高危型人乳头瘤病毒(HR-HPV)DNA检测阴性的女性发生子宫颈癌前病变及子宫颈癌的风险显著低于细胞学检查结果正常者^([1,2]),因此,全球多国已逐步将子宫颈癌筛查策略调整为基于人乳头瘤病毒(HPV)的初筛。然而,单纯HPV感染并不等同于疾病状态,约90%的感染可在3年内自然清除,仅少数持续感染最终进展为癌前病变及癌^([3])。因此,需通过分流进行风险分层避免过度诊疗。尽管子宫颈脱落细胞学检查具有高特异性,是目前HPV初筛阳性女性分流管理的主要手段,但是基于单一形态学标准的风险分层可能会高估部分低危风险型别HPV感染的临床意义。

关键词： 养胃颗粒   幽门螺杆菌   三联疗法   铋剂四联疗法   慢性胃炎   消化性溃疡   meta分析  

摘要： 目的:系统评价养胃颗粒辅助三联疗法(triple therapy,TR)/铋剂四联疗法(bismuth quadruple therapy,BQT)治疗幽门螺杆菌(Helicobacter pylori,HP)阳性胃炎及消化性溃疡的有效性及安全性。方法:计算机检索CNKI、万方、维普、CBM、PubMed、Embase和Cochrane library数据库,搜集养胃颗粒联合TR/BQT治疗HP阳性胃炎及消化性溃疡的随机对照试验(RCT),检索时间均从建库至2024年2月29日,根据纳入与排除标准选择2名研究人员进行文献筛选和资料提取,同时对纳入研究进行偏倚风险评价,采用RevMan 5.3及Stata 17.0软件进行meta分析。结果:共纳入17个RCT、2030例患者,meta分析结果显示:与TR/BQT比较,养胃颗粒联合TR/BQT在HP根除率(RR=1.18,95%CI:1.13~1.23,P<0.001)、临床总有效率(RR=1.20,95%CI:1.14~1.26,P<0.001)、提高血清胃泌素水平(RR=1.30,95%CI:0.77~1.84,P<0.001)、降低血清内皮素水平(RR=-4.13,95%CI:-5.93~-2.32,P<0.001)以及降低血清胃动素水平(RR=-11.78,95%CI:-16.30~-7.26,P<0.001)方面均优于对照组,均差异有统计学意义;同时可以显著提高胃蛋白酶原Ⅰ水平(RR=7.45,95%CI:5.37~9.53,P<0.001)及胃蛋白酶原比值(RR=0.73,95%CI:0.51~0.95,P<0.001),均差异有统计学意义;在不良反应发生率(RR=0.82,95%CI:0.46~1.46,P=0.50)以及降低胃蛋白酶原Ⅱ水平(RR=-0.23,95%CI:-1.11~0.64,P=0.60)方面差异无统计学意义。结论:与TR或BQT方案相比,养胃颗粒在HP根除率、临床总有效率方面均优于对照组,同时可改善HP阳性胃炎及消化性溃疡患者胃肠激素、血管活性肽及胃蛋白酶原水平,并且在治疗过程中没有明显增加不良反应发生率。

关键词： 糖尿病患者   糖尿病肾脏病   早期信号  

摘要： 糖尿病肾脏病是糖尿病常见的并发症,也是糖尿病患者应该重视预防的。糖尿病肾脏病发展缓慢,往往没有明显的症状,早期诊断和干预可以有效防止病情的进一步恶化。糖尿病患者及其家属应当了解肾病的早期信号,以便及早发现、及时治疗,减少并发症的发生。本文将介绍糖尿病肾脏病的早期信号,并提供一些防治建议,帮助糖尿病患者保护肾脏健康。

关键词： 丹红注射液   动脉粥样硬化   炎症   机制  

摘要： 目的采用生物信息学的方法分析丹红注射液干预动脉粥样硬化炎症反应的潜在机制。方法运用TCMSP数据库获得丹红注射液成分信息,预筛选丹红注射液的化学小分子对应的靶蛋白。利用STRING数据库对靶蛋白进行蛋白质互作(PPI)分析。基于R包对靶蛋白进行基因本体论(GO)功能、京都基因与基因组百科全书(KEGG)通路富集分析,靶蛋白进行疾病本体论(DO)富集分析。在CTD数据库中以“Atherosclerosis”为关键词搜索动脉粥样硬化相关基因,选取推理得分≥100的靶标与上一步所预测的靶蛋白取交集进行筛选以缩小靶蛋白范围。对目标靶蛋白进行GO功能、KEGG通路富集分析,利用STRING数据库对靶蛋白进行PPI分析,使用Cytoscape软件对其进行网络图构建,通过Cytohubba插件计算MCC得分筛选出关键的前10靶标。利用Cytoscape构建中药-成分-靶蛋白-通路药理学网络,通过Autodock软件研究目标蛋白与小分子之间的对接稳定性。结果丹红注射液的中药成分为红花和丹参,11个活性成分分子。筛选得到与炎症、动脉粥样硬化相关的有效活性成分7个小分子,10个hub基因,选取交集靶标相关的2个中药成分、7个分子、69个靶标、81条通路构建药理学网络,网络共159个节点,874条关系对。丹参酚醌I、新隐丹参酮与白细胞介素6(IL-6)、IL-1β、肿瘤坏死因子(TNF)分子对接稳定。结论丹红注射液对动脉粥样硬化的潜在机制涉及多个关键靶点和多条信号通路,各靶点与各通路之间产生的交叉作用,可能会抑制炎症反应,为其在动脉粥样硬化性心脑血管疾病中的应用提供参考依据。

关键词： Bioactive equivalent combinatorial   components   Xiao-Xu-Ming decoction   Coronary artery spasm   Calmodulin   MLCK/MLC   Vascular smooth muscle cells  

摘要： Background: Coronary artery spasm (CAS) is a severe pathological disorder with limited treatment options. Inhibiting vascular smooth muscle cell (VSMC) proliferation has emerged as a promising therapeutic strategy for CAS. Purpose: This study aims to investigate the potential function of Xiao-Xu-Ming decoction (XXMD) in the attenuation of CAS in rats. Methods: The involvement of bioactive equivalent combinatorial components (BECCs) of XXMD in CAS was predicted using network pharmacological analysis. A CAS rat model was established using pituitrin (Pit), and an Ang II-induced cell model was developed to assess the effects of BECCs on VSMC contraction. The expression of contractile phenotype markers was analyzed using RT-qPCR and Western blotting. Additionally, MTT assay, flow cytometry, Annexin V/PI staining, and Transwell assay were performed to evaluate cell proliferation, cell cycle progression, apoptosis, and migration. A collagen gel contraction assay was conducted to assess VSMC contraction. Results: Network pharmacological analysis suggested that BECCs in XXMD may influence CAS development through the Calmodulin (CaM)-MLCK/MLC axis. The drug-containing XXMD serum significantly inhibited VSMC proliferation and migration by reducing the expression of MLCK, alpha-SMA, Calponin, and SMHHC. Furthermore, it suppressed VSMC proliferation by downregulating MLCK expression via CaM inhibition. Importantly, BECCs of XXMD promoted anti-inflammatory responses, vasorelaxation, and oxidative stress reduction in Pit-induced CAS rats. Conclusion: These findings suggest that BECCs in XXMD may counteract CAS by modulating the CaM-mediated MLCK/MLC pathway in VSMCs. This mechanism may offer a foundation for novel and effective therapeutic strategies to treat CAS and prevent associated cardiac events, such as myocardial infarction and sudden cardiac death. Novelty: This study identifies a novel mechanism by which the "active ingredient cluster" of XXMD alleviates CAS through modul

关键词： calmodulin   calmodulin-kinase cascade   CaMKK alpha/1   oligomerization   protein-protein interaction   regulatory domain  

摘要： Calcium/calmodulin-dependent protein kinase kinase (CaMKK) is an activating kinase for calcium/calmodulin-dependent protein kinase type 1 (CaMKI), calcium/calmodulin-dependent protein kinase type IV (CaMKIV), RAC-alpha serine/threonine-protein kinase (PKB), and AMP-activated protein kinase (AMPK) that has been reported to form an active oligomer in cells. Glutathione S-transferase (GST) pulldown assay from the extracts of COS-7 cells expressing GST- and His6-CaMKK alpha/1 mutants showed that the C-terminal region containing the autoinhibitory and calmodulin (CaM)-binding sequence (residues 438-463) is required for CaMKK alpha/1 homo-oligomerization. This was confirmed by the fact that the GST-CaMKK alpha/1 C-terminal domain (residues 435-505) directly interacted with EGFP-CaMKK alpha/1 residues 435-505 as well as with wild-type CaMKK alpha/1. Notably, once oligomerized in cells, CaMKK alpha/1 is neither exchangeable between the oligomeric complexes nor dissociated by Ca2+/CaM binding. These results support stable oligomerization of CaMKK in the cells by intermolecular self-association of its C-terminal region containing a regulatory domain.

关键词： p53   MDM2   PP2A   IER5   Phosphorylation   Ubiquitination  

摘要： The tumor suppressor p53 activates transcription of the IER5 gene, which encodes an adapter protein of protein phosphatase PP2A. IER5 binds to both the B55 regulatory subunit of PP2A and PP2A's target proteins, facilitating PP2A/B55-catalyzed dephosphorylation of these proteins. Here, we show that IER5 functions as a positive regulator of p53 by inhibiting its ubiquitination, thereby increasing cellular p53 levels. Mechanistically, this effect of IER5 requires its nuclear localization and binding to both PP2A/B55 and the p53 ubiquitin E3 ligase MDM2. Importantly, IER5 fails to inhibit p53 ubiquitination in cells treated with the MDM2 inhibitor Nutlin-3. The IER5-PP2A/B55 complex dephosphorylates MDM2 at Ser166, leading to MDM2 ubiquitination and a reduction in nuclear MDM2. Altogether, our data provide evidence that IER5-PP2A/B55 regulates the nuclear balance between MDM2 and p53 via MDM2 dephosphorylation.

关键词： calcium   calmodulin   EGFR   FBXL2   protein degradation   NSCLC   EGFR-TKI resistance  

摘要： Epidermal growth factor receptor (EGFR)-targeted therapy by tyrosine kinase inhibitors (TKIs) is the first-line treatment of non-small cell lung cancer (NSCLC). However, EGFR mutation-mediated TKIs resistance remains a major hurdle for NSCLC treatment. This study found the opposite roles of calcium (Ca2+) and its receptor calmodulin (CaM) in regulating EGFR protein stability. Elevated Ca2+ facilitates degradation of EGFR TKI-resistant mutant proteins, whereas CaM protects them from degradation. Mechanistically, Ca2+ binding to CaM triggers heat shock protein 70-mediated lysosomal degradation of CaM protein, resulting in reduced CaM binding to EGFR, thereby facilitating EGFR proteasomal degradation mediated by the E3 ubiquitin ligase FBXL2. Notably, a combination of Ca2+ activator curcumin, FBXL2 activator nebivolol, and osimertinib significantly inhibits EGFRT790M/C797S-driven TKI-resistant NSCLC growth. Together, this study highlights the importance of the negative feedback loop between Ca2+ and CaM and the critical regulatory role of Ca2+/CaM in the FBXL2-mediated EGFR degradation, providing a viable therapeutic strategy for TKI-resistant NSCLC.

关键词： calcium/calmodulin dependent kinase II   calmodulin   epilepsy   hyperexcitability   Na(V)1.2 channel  

摘要： Objective Dysfunction of calcium/calmodulin (CaM)-dependent kinase II (CaMKII) has been involved in hyperexcitability-related disorders including epilepsy. However, the relationship between CaMKII and neuronal excitability remains unclear. Methods Neuronal excitability was detected in vivo and in vitro by electroencephalography (EEG), patch clamp and multi-electrode array (MEA), respectively. Next, we assessed the currents of voltage-gated sodium channels (VGSCs) by patch clamp, and mRNA and protein expressions of VGSCs were determined by real-time quantitative reverse transcription-polymerase chain reaction (qRT-PCR) and western blot, respectively. Meanwhile, the association between the nuclear receptor subfamily 4 group A member 2 (NR4A2) and promoters of Scn2a, was determined by chromatin immunoprecipitation (ChIP)-qPCR. In addition, we utilized co-immunoprecipitation (Co-IP), immunofluorescence labeling, and pull-down to determine the interaction between VGSCs and CaM. Results Prolonged CaMKII inhibition by KN93, an inhibitor of CaMKII, for 24 h and CaMKII knockdown induced more seizure-like events in Wistar rats, TRM rats and C57BL/6 mice, and led to hyperexcitability in primary hippocampal neurons and human induced-pluripotent stem cell (hiPSC)-derived cortical neurons. In addition, prolonged CaMKII inhibition resulted in elevated persistent sodium current (INaP)/transient sodium current (INaT) and increased mRNA and protein expressions of NaV1.2. Meanwhile, prolonged CaMKII inhibition by KN93 decreased NR4A2 expression and contributed to a transcriptional repression role of NR4A2 in Scn2a regulation, leading to increased NaV1.2 expression. Moreover, an increased interaction between NaV1.2 and CaM was attributable to enhanced binding of CaM to the isoleucine-glutamine (IQ) domain at the C-terminus of the NaV1.2 channel, which may also lead to the potentiation in INaP/INaT and channel activity. Furthermore, a peptide that antagonized CaM binding to NaV1.2 IQ do

关键词： 麸炒苍术   粗多糖   精制多糖   免疫抑制   肠道损伤   代谢组学   亚油酸代谢  

摘要： 以环磷酰胺诱导的免疫抑制和肠道损伤小鼠为模型，评价麸炒苍术精制多糖改善免疫抑制小鼠肠道损伤的药效优势，并探讨其作用机制。50只C57BL/6雄性小鼠随机分为正常组、模型组、麸炒苍术粗多糖组(1200 mg·kg-1)、麸炒苍术精制多糖组(842 mg·kg-1)、阳性药组(盐酸左旋咪唑40 mg·kg-1)，每组10只，正常组灌胃生理盐水，其他组灌胃相应药物，连续15 d，除正常组外，其余各组小鼠在第8～10天腹腔注射环磷酰胺(80 mg·kg-1)构建免疫抑制和肠道损伤小鼠模型。实验过程中监测小鼠体质量，实验结束后测定小鼠脾脏和胸腺指数；采用苏木精-伊红（HE）染色评估脾脏、胸腺及小肠组织的病理变化；采用实时荧光定量PCR（qRT-PCR）检测小肠组织中肿瘤坏死因子-α（TNF-α）、白细胞介素-6（IL-6）、白细胞介素-1β（IL-1β）的mRNA表达水平；通过阿利新蓝-过碘酸-雪夫染色（AB-PAS）、免疫组化染色检测小肠组织中杯状细胞数量、黏蛋白2（MUC2）的表达；通过免疫荧光检测小肠组织中紧密连接蛋白-1（ZO-1）、闭合蛋白（occludin）和免疫球蛋白A（IgA）分泌细胞的表达；采用酶联免疫吸附测定法（ELISA）检测小肠组织中分泌型免疫球蛋白A（sIgA）的含量；通过非靶向代谢组学检测小鼠小肠组织代谢物变化。结果显示，与模型组小鼠相比，麸炒苍术粗多糖组和麸炒苍术精制多糖组小鼠体质量、胸腺指数、脾脏指数、杯状细胞数量、MUC2、TNF-α、IL-6、IL-1β、ZO-1、occludin和IgA分泌细胞、sIgA的表达显著升高。麸炒苍术精制多糖对胸腺指数、ZO-1、occludin、TNF-α、IL-1β、IL-6的调控能力显著强于粗多糖。代谢组学分析表明，模型组、麸炒苍术粗多糖组、麸炒苍术精制多糖组鉴定出13个差异代谢物，代谢途径涉及不饱和脂肪酸生物合成代谢、二羧酸代谢、亚油酸代谢等15条代谢途径，其中亚油酸代谢为麸炒苍术多糖调节的主要代谢途径。与麸炒苍术粗多糖相比，精制多糖对小肠组织代谢物具有更好的调节能力。综上，该研究表明麸炒苍术粗多糖和精制多糖均能改善环磷酰胺诱导的免疫抑制和肠道损伤，且精制多糖药效更优，其作用机制与亚油酸代谢通路密切相关。