Objectives:COVID-19 has been identified as causing damage to the nervous and endocrine systems, with observed effects on hypopituitarism. However, the effects of COVID-19 on hypopituitarism are controversial. In this study, we conducted MR analysis to investigate the relationship between COVID-19 and hypopituitarism. Study design:The MR study utilized publicly available summary statistics from an independent European cohort. Data were sourced from the EBI database for COVID-19(case N = 8316 and control N = 1549,095) and from the FinnGen database for hypopituitarism (case N = 910 and control N = 211,123). Independent genetic variants significantly associated with each exposure(P < 5 ×10-8) were considered as instrumental variables. The MR analysis and colocalisation analysis were underwent. Methods:Four methods were employed: IVW, MR-Egger regression, weighted median and weighted mode with sensitivity analyses conducted. Finally, colocalisation analysis was also conducted. Results:MR study analyses were performed using the IVW fixation method, and we found a significant causal association between COVID-19 and hypopituitarism. IVW OR= 1.283, 95 %CI(1.017, 1.617), p = 0.035; MR-Egger OR= 1.355, 95 % CI(0.850, 2.160), p = 0.291; OR= 1.327, 95 %CI(1.017, 1.732), p = 0.037 in the weighted median approach and OR= 1.330, 95 % CI(0.993, 1.782), p = 0.128 in the weighted modal approach, with the results of the sensitivity analysis showing a stable and suggestive of a causality that COVID-19 increases the risk of developing hypopituitarism. The probability of colocalization of COVID-19 and hypopituitarism was PP.H4.abf= 9.55 %. Conclusion:In this study, we have established a causal relationship between COVID-19 and the risk of hypopituitarism. This suggests that vigilance against the development of hypopituitarism is warranted following SARS-CoV-2 infection, and underscores the potential for long-term autonomic dysfunction in post-COVID-19 clinical management.

Background:Alzheimer's disease (AD) is a chronic neurodegenerative disease and the most prevalent form of dementia. Fenugreek seeds possess anti-inflammatory and antioxidant effects, making them valuable therapeutic agents in managing neurodegenerative diseases. Objective:This study aimed to investigate the primary biological pathways and specific mechanisms underlying the protective effects of fenugreek in preventing mice of AD by employing bioinformatics and experimental verification. Methods:We administered fenugreek extract as an intervention in mice model of AD and then assessed their cognitive ability and histopathological changes. We predicted the key target genes associated with fenugreek action on AD and the main biological pathways using the bioinformatics method. Furthermore, we observed the different expression of target genes by western blot (WB). Results:The bioinformatics analysis revealed a strong correlation between fenugreek and AD. The behavioral experiments confirmed that fenugreek improved the behavioral and cognitive dysfunction in mice with AD. The histopathology revealed significant changes that fenugreek can inhabit Nissl bodies. Western blot experiments confirmed that fenugreek exerted statistically significant modulatory effects on the levels of inflammatory proteins [interleukin-6 (IL-6), IL-10, and IL-1β] and oxidative stress-related proteins (amyloid-β protein precursor, apolipoprotein E, and presenilin 1). Conclusions:This study suggested that fenugreek might be involved in the AD pathway and effectively prevented the progression of AD through significant anti-inflammatory and antioxidant effects.

Focal adhesion kinase (FAK) belongs to the nonreceptor tyrosine kinases, which selectively phosphorylate tyrosine residues on substrate proteins. FAK is associated with bladder, esophageal, gastric, neck, breast, ovarian and lung cancers. Thus, FAK has been considered as a potential target for tumor treatment. Currently, there are six adenosine triphosphate (ATP)-competitive FAK inhibitors tested in clinical trials but no approved inhibitors targeting FAK. Defactinib (VS-6063) is a second-generation FAK inhibitor with an IC50 of 0.6 nM. The binding model of VS-6063 with FAK may provide a reference model for developing new antitumor FAK-targeting drugs. In this study, the VS-6063/FAK binding model was constructed using ensemble docking and molecular dynamics simulations. Furthermore, the molecular mechanics/generalized Born (GB) surface area (MM/GBSA) method was employed to estimate the binding free energy between VS-6063 and FAK. The key residues involved in VS-6063/FAK binding were also determined using per-residue energy decomposition analysis. Based on the binding model, VS-6063 could be separated into seven regions to enhance its binding affinity with FAK. Meanwhile, 60 novel defactinib-based compounds were designed and verified using ensemble docking. Overall, the present study improves our understanding of the binding mechanism of human FAK with VS-6063 and provides new insights into future drug designs targeting FAK. Communicated by Ramaswamy H. Sarma

Abstract Alzheimer's disease (AD), a neurodegenerative disease, has a complex pathological mechanism involving oxidative stress, endoplasmic reticulum (ER) stress and other pathways. Thioredoxin 1 (Trx-1), the major redox regulator, may be an effective treatment strategy for AD. Therefore, the present study explored the role and possible mechanism of Trx-1 in AD. The expression of Trx-1 in the hippocampus of AD was verified by WB and RT-PCR, and the effects of Trx-1 on behavioral function and neuropathological damage of AD mice were analyzed by open field test, water maze experiment, HE staining, and protein expression analysis of amyloid β-protein (Aβ), Tau protein (Tau) and p-Tau. At the same time, oxygen species (ROS) and ER stress were detected to further analyze the mechanism. Trx-1 expression in the hippocampus of AD model mice was reduced, and the oe-Trx-1 remarkably enhanced it. We observed an obvious behavioral cognitive dysfunction and neuropathological damage in AD model mice. Trx-1 significantly ameliorated the anxiety-like behavior and cognitive ability in AD mice, alleviated the pathological damage of hippocampal tissue, and reduced the protein expressions of Aβ, Tau, and p-Tau, suggesting that Trx-1 could alleviate behavioral cognitive dysfunction and neuropathological damage in AD. In addition, Trx-1 significantly reduced ROS levels and the GRP78, PERK, IRE1α, and CHOP protein expression and relieved ER stress in AD mice. Trx-1 may alleviate behavioral cognitive dysfunction and neuropathological damage in AD mice by regulating ER stress.

Alzheimer's disease (AD) is the most commonly seen neurodegenerative brain disorder. The paracrine effects of mesenchymal stem cells (MSCs) signify to trigger immunomodulation and neural regeneration. However, the role and mechanism of bone marrow MSC- (BMSC-) derived CX3CL1 in AD remains elusive. In this study, Aβ1-42-intervened SH-SY5Y cells were used for AD cell model construction. pcDNA-ligated CX3CL1 overexpression plasmids were transfected into BMSCs. The levels of soluble and membrane-bound CX3CL1 were detected by ELISA and Western blotting (WB), respectively. The growth, apoptosis, and pathology of AD model cells were evaluated by CCK-8, flow cytometry, immunofluorescence, morphology observation, biochemical examination, and WB. It was found that Aβ1-42 significantly reduced CX3CL1 expression either in soluble or membrane-bound form, cell viability, relative protein expression of synaptic markers, SOD, CAT, and GSH-Px contents, as well as Trx protein expression; in addition, it enhanced the apoptosis rate, the relative expression of cleaved caspase-3, Aβ, tau, p-Tau, Iba1, MDA, TXNIP, and NLRP3 in SH-SY5Y cells; however, the above effects were prominently reversed by the coculture of BMSCs. Moreover, overexpression of CX3CL1 in BMSCs observably strengthened the corresponding tendency caused by BMSCs. In conclusion, through the TXNIP/NLRP3 pathway, CX3CL1 derived from BMSCs inhibited pathological damage in Aβ1-42-induced SH-SY5Y.

目的 探讨飞燕草素对光化学损伤661W细胞的保护作用及其机制.方法 取661W细胞进行培养,根据预实验结果采用(2000±200)lux光照强度持续照射细胞48 h作为造模条件,采用5 μmol·L-1飞燕草素、3 mmol·L-1抗氧化剂N-乙酰-L-半胱氨酸(NAC)为最佳用药浓度.细胞分组如下:对照组,常规避光培养48 h;光照组,(2000±200)lux光照培养48 h;光照飞燕草素组,(2000±200)lux光照培养24 h,换含5 μmol.L-1飞燕草素的培养基继续光照培养24 h;避光飞燕草素组,避光培养24 h,换含5 μmol·L-1飞燕草素的培养基继续避光培养24 h;光照NAC组,(2000±200)lux光照培养24 h,换含3 mmol·L-1 NAC的培养基继续光照培养24 h.采用显微镜观察各组细胞状态、CCK-8 检测细胞生存率、DCFH-DA荧光探针染色检测细胞活性氧(ROS)水平、JC-10 染色检测线粒体膜电位、Annexin V-FITC/PI染色检测细胞凋亡率、Western blot检测氧化应激线粒体凋亡通路相关蛋白表达水平.结果 显微镜下可见,对照组细胞生长状态良好;光照组细胞皱缩卷曲,脱落细胞增加.CCK-8检测结果显示:与对照组相比,光照组细胞生存率均明显降低,差异有统计学意义(P＜0.05);避光飞燕草素组细胞生存率无明显变化(P＞0.05).与光照组相比,光照飞燕草素组、光照NA C组和避光飞燕草素组细胞生存率均明显回升,差异均有统计学意义(均为P＜0.05).与对照组相比,光照组ROS含量均明显上升,差异有统计学意义(P＜0.05);避光飞燕草素组RO S含量减少不明显,差异无统计学意义(P＞0.05).与光照组相比,光照飞燕草素组、光照NAC组和避光飞燕草素组ROS含量均明显下降,差异均有统计学意义(均为P＜0.05).与对照组相比,光照组线粒体膜电位均明显下降,差异有统计学意义(P＜0.05);避光飞燕草素组线粒体膜电位变化不明显,差异无统计学意义(P＞0.05).与光照组相比,光照飞燕草素组、光照NAC组和避光飞燕草素组线粒体膜电位明显上升,差异均有统计学意义(均为P＜0.05).与光照组相比,光照飞燕草素组、光照NAC组和避光飞燕草素组细胞凋亡率均明显降低,差异均有统计学意义(均为P＜0.05).与光照组相比,光照飞燕草素组、光照NAC组和避光飞燕草素组的iNOS、Bax、细胞色素C、Cleaved-Caspase-3蛋白表达均下调,Bcl-2蛋白表达均上调,Bcl-2/Bax值均明显上升,差异均有统计学意义(均为P＜0.05).结论 飞燕草素可通过调节氧化应激线粒体凋亡通路相关蛋白表达,对光化学损伤661W细胞产生保护作用.

背景:临床药物单独使用对脊髓损伤特定病理变化具有较好地阻断作用,但单一用药往往存在较为复杂的不良反应,如雌激素可以缓解氧化损伤导致的细胞凋亡,但同时会激活体内的炎症反应.在此大背景下,联合协同用药成为临床创新发展的趋势,其联合用药机制也迫切需要阐明.目的:探究地塞米松联合雌激素治疗对脊髓钝挫伤大鼠白细胞介素6、Caspase3和Bcl-2表达的影响.方法:雌性4周龄SD大鼠48只,随机分为假手术组、生理盐水组、地塞米松组、联合治疗组,每组12只.假手术组显露脊髓后直接缝合,其他3组采用改良Allen's法建立脊髓损伤模型,术后30 min和1,2,3,4,5d时假手术组和生理盐水组大鼠注射生理盐水(0.2 mL/kg),地塞米松组大鼠注射地塞米松(0.2 mg/kg),联合治疗组大鼠先后注射雌二醇(0.04 mg/kg)、地塞米松(0.2 mg/kg),注射部位均为大鼠前肢大腿外侧肌.术后14d内采用BBB评分观察大鼠后肢运动功能恢复情况,术后3,7,21d采用免疫组织化学检测脊髓组织中白细胞介素6、Bcl-2和Caspase3的表达.结果 与结论:①联合治疗组运动功能恢复开始时间较地塞米松组和生理盐水组更早,在整个恢复时期,联合治疗组的BBB评分显著高于地塞米松组(P＜0.05);②与地塞米松组相比,联合治疗组白细胞介素6和Casepase3表达显著降低、Bcl-2表达明显升高(P＜0.05);③结果表明,联合治疗能有效减少脊髓损伤后促凋亡因子Caspase3和促炎因子白细胞介素6的表达,增加抗凋亡因子Bcl-2的表达,通过抑制凋亡和炎症促进脊髓损伤后功能恢复.

Abstract Background: Spinal cord contusion (SCC) results in a series of pathophysiologic consequences such as edema, apoptosis, and inflammation. However, inflammation may also be beneficial for the recovery of motor function after SCC, but the underlying mechanisms remain incompletely elucidated. Interleukin-1 beta (IL-1β) is a pro-inflammatory factor that has synergistic effects with other inflammatory factors to aggravate spinal cord injury. Inflammatory factors have been found to activate the serine/threonine-specific protein kinase, protein kinase B (AKT) and to inhibit cell survival, but it is not clear whether inflammation upregulates the expression of IL-1β in the rat model of SCC and subsequently interferes in the phosphatidylinositol-3-kinase (PI3K)/AKT signaling pathway. Therefore, this study explored whether IL-1β affects the recovery of motor function in spinal cord injury by interfering with the PI3K/AKT signaling pathway. Method: SCC rats were established by the Allen method. The Basso Beattie Bresnahan (BBB) scoring was used to assess motor function in the spinal cord of injured rats. Quantitative polymerase chain reaction and Western blot were used to determine the expression of genes and proteins of IL-1β, PI3K, and AKT1. Immunohistochemistry and immunofluorescence were used to locate and detect IL-1β and AKT1 proteins in spinal cord tissue. To further explore the underlying mechanism of IL-1β, lentivirus was constructed by RNA interfering (RNAi) technique to inhibit the expression of IL-1β, and bioinformatics was applied to show the relationship between IL-1β and AKT1. Results: BBB scores decreased after SCC, and IL-1β and AKT1 was located in the cytoplasm of spinal cord anterior horn neurons. In the early stage of SCC, the expression level of IL-1β gene and protein in the experimental group was higher than that in the sham operated group. At the same time, expression of the AKT1 gene decreased. After expression of IL-1β mediated by lentivirus was inhibited, BBB scores increased significantly, and spinal cord motor function improved. Bioinformatic analysis revealed a relationship between IL-1β and AKT1. In addition, AKT1 gene expression was upregulated and PI3K expression was unchanged in the PI3K/AKT signaling pathway. Conclusion IL-1β not only exacerbates the inflammatory response after SCC, but also interferes with motor function. Inhibition of IL-1β may promote recovery of spinal cord injury by upregulating AKT1 in the PI3K/AKT signaling pathway, which provides a new perspective for future clinical practice in treating spinal cord injury