关键词： 癌症   癌症复发恐惧   心理干预   网状meta分析  

摘要： 目的:分析不同心理干预对癌症患者癌症复发恐惧(FCR)的影响。方法:检索中英文献数据库中关于不同心理干预对癌症患者FCR影响的随机对照试验,采用网状meta分析探讨干预效果,以标准化均数差(SMD)、95%置信区间(CI)和累积排序概率图下面积(SUCRA)为效应指标。结果:共纳入34项研究(n=3772),包括接纳承诺疗法(ACT)、感恩拓展干预(GET)、认知行为疗法(CBT)、正念疗法(MT)和综合心理干预(Mul)5种心理干预,根据效果评价时间将其分为干预后即刻组、干预后1~2月组、干预后3~6月组。与常规护理相比,干预后即刻组中,ACT(SMD=-1.80,95%CI:-2.47~-1.13)、GET(SMD=-1.33,95%CI:-2.07~-0.59)、MT(SMD=-0.59,95%CI:-1.03~-0.14)和CBT(SMD=-0.54,95%CI:-0.93~-0.15)可有效降低FCR,ACT的SUCRA值最大(0.96);干预后1~2月组中,GET(SMD=-2.32,95%CI:-2.99~-1.65)、ACT(SMD=-1.46,95%CI:-2.23~-0.70)可有效降低FCR,GET的SUCRA值最大(0.99);干预后3~6月组中,Mul(SMD=-1.82,95%CI:-3.03~-0.61)和MT(SMD=-1.43,95%CI:-2.33~-0.54)可有效降低FCR,Mul的SUCRA值最大(0.90)。结论:不同心理干预在不同效果评价时间点对FCR的效果不同,临床应结合患者特点和资源选择适宜方法。

关键词： 糖尿病足溃疡   中药塌渍法   富血小板血浆   清创   负压持续吸引   细菌转阴率   炎症因子水平  

摘要： 目的探讨中药塌渍法联合富血小板血浆(PRP)治疗在糖尿病足溃疡(DFU)中的应用效果。方法按随机数表法将于我院治疗的82例DFU患者分为两组,病例选取时间为2022年1月-2023年12月,每组各41例。对照组采用常规法治疗,于对照组基础上,观察组加用中药塌渍法联合PRP治疗。比较两组细菌转阴率、炎症因子水平[白细胞(WBC)、C反应蛋白(CRP)及红细胞沉降率(ESR)]、疼痛评分、二期修复准备时间、创面愈合时间、住院时间及复发情况。结果较对照组,观察组治疗后细菌培养转阴率更高(P<0.05)。较对照组,观察组治疗后WBC、CRP、ESR水平及视觉模拟评分法(VAS)评分更低,二期修复准备时间、创面愈合时间及住院时间更短(P<0.05)。较对照组,观察组复发率更低(P<0.05)。结论中药塌渍法联合PRP治疗DFU可加速创面的再上皮化,缓解患者疼痛,提高细菌转阴率,降低WBC、CRP、ESR水平,缩短二期修复准备时间及创面愈合时间,减少疾病复发。

关键词： IgA肾病   麻黄连翘赤小豆汤   炎症反应   补体激活   C3a/C3aR信号通路   大鼠  

摘要： 目的基于C3a/C3aR信号通路探讨麻黄连翘赤小豆汤对IgA肾病大鼠炎症反应的影响及作用机制。方法30只雄性Wistar大鼠随机选取9只作为对照组,其余21只采用牛血清白蛋白-脂多糖-四氯化碳复合免疫法制备IgA肾病大鼠模型。将造模成功的18只大鼠随机分为模型组、中药组和西药组。中药组予麻黄连翘赤小豆汤混悬液灌胃,西药组予氯沙坦钾混悬液灌胃,对照组和模型组予生理盐水灌胃,连续4周。HE、PAS、Masson染色观察肾组织形态,免疫荧光染色检测肾小球系膜区IgA表达,ELISA检测肾组织白细胞介素(IL)-6、肿瘤坏死因子-α(TNF-α)、C3a含量,Western blot检测肾组织C3/C3b/C3c、C3aR、补体B因子(CFB)、Toll样受体4(TLR4)、核因子-κB(NF-κB)p65蛋白表达,免疫组化检测肾组织C3、C3aR、CFB阳性表达。结果与对照组比较,模型组大鼠肾小球扩张、系膜细胞增生,系膜基质扩张,肾小球系膜区IgA免疫复合物沉积明显增多(P<0.01),肾组织IL-6、TNF-α、C3a含量明显升高(P<0.01),肾组织C3/C3b/C3c、C3aR、CFB、TLR4、NF-κBp65蛋白表达明显升高(P<0.05),C3、C3aR、CFB阳性表达明显升高(P<0.01);与模型组比较,中药组和西药组大鼠肾组织病理损伤减轻,IgA免疫复合物沉积明显减少(P<0.01),肾组织IL-6、TNF-α含量明显降低(P<0.01,P<0.05),C3/C3b/C3c、C3aR、CFB、TLR4、NF-κBp65蛋白表达降低(P<0.01,P<0.05),C3、C3aR、CFB阳性表达明显降低(P<0.01)。结论麻黄连翘赤小豆汤可抑制IgA肾病大鼠炎症反应,改善肾组织病理损伤,其作用机制可能与抑制替代途径补体激活,调控C3a/C3aR和TLR4/NF-κB信号通路有关。

关键词： Schistosoma   calcium/calmodulin-dependent protein kinase   II (CaMKII)   sarco/endoplasmic reticulum calcium-ATPase   SERCA   praziquantel resistance   calcineurin   P-glycoprotein (P-gp)  

摘要： Calcium/calmodulin-dependent protein kinase II (CaMKII) performs diverse essential functions through integrating a range of calcium signals. In Schistosoma, two Calmodulin (CaM) genes are characterized. CaMKII exhibits distinct expression patterns across the developmental stages of the parasite. Its significance lies in sustaining Schistosoma survival and maintaining calcium homeostasis. As it is a calcium sensing protein, its function is closely related to the efficacy of praziquantel, the mainstay drug against schistosomiasis. The relationship between CaMKII and praziquantel involves several potential factors. Praziquantel induces an increased calcium influx into Schistosoma that binds CaM and activates CaMKII, which in turn mitigates the effect of the drug and potentially contributes to praziquantel resistance in several ways. By maintaining calcium homeostasis, CaMKII opposes the surge in calcium influx induced by praziquantel. It modulates voltage-gated calcium channels and reduces calcium influx. It also inhibits ryanodine receptors and inositol triphosphate receptors, thus preventing the release of calcium from the sarcoplasmic/endoplasmic reticulum. CaMKII activates nuclear factor kappa B and subsequently activates sarco/endoplasmic reticulum calcium-ATPase (SERCA), which increases calcium uptake into the sarcoplasmic/endoplasmic reticulum and decreases cytosolic calcium. Nuclear factor-kappa B, activated by CaMKII may lead to up-regulation of P-glycoprotein, which facilitates praziquantel efflux. CaMKII also activates calcineurin that inhibits SERCA. Given its pivotal role in Schistosoma homeostasis and survival, CaMKII emerges as a promising target for novel anthelmintic therapies, and its modulation might enhance the efficacy of praziquantel.

关键词： Antimicrobial peptides   Defensin   Fusion protein   Calmodulin   Escherichia coli   Recombinant production   Disulfide bonds  

摘要： Human alpha-defensin 5 (HD5), a cysteine-rich antimicrobial peptide critical for intestinal innate immunity, has been extensively studied for its structural and functional properties. Both the reduced form (HD5red) and the oxidized form (HD5oxi) exist in vivo and exhibit distinct antimicrobial activity spectra. In this study, we developed an efficient method to overexpress recombinant HD5 in Escherichia coli (E. coli) BL21 (DE3) strain by using calmodulin (CaM), which also interacts with antimicrobial peptides, as a fusion partner. Fusion expression suppressed the degradation of HD5 and reduced its toxicity to host cells. Following purification of the fusion protein and enzymatic cleavage to release the HD5 region, we successfully obtained sufficient amounts (yielding 1.5-1.7 mg/L culture) of active recombinant HD5oxi and HD5red for various applications, including stable isotope-labeled peptides for NMR analysis. Furthermore, we investigated the protective effect of CaM fusion and the mechanism of disulfide bond formation using CD and NMR spectroscopy, structural prediction, and molecular dynamics simulations. Our results suggest that the appropriate interaction strength between CaM and the HD5 region in the fusion state is a key factor for stable production.

关键词： Foxtail millet   CAMTA   Transcription factor   Protein interaction   Codon bias   Evolution  

摘要： Calmodulin-binding transcriptional activator (CAMTA) is a calmodulin-binding transcription factor, which plays an important role in calcium/calmodulin transduction signaling pathway. Foxtail millet (Setaria italica L.) is an economically important C4 model crop, but the genome-wide identification and molecular evolution of the CAMTA family is not yet available. In this study, seven CAMTA genes were identified from the foxtail millet genome via bioinformatics methods. They were overall similar in structure but differed, and all showed tissue expression specificity. Regarding molecular evolution, codon bias contributed to the evolution of the CAMTA family, and interestingly, base mutation was not the main factor affecting their codon bias, and may also be affected by factors such as natural selection and other factors. Multiple traces indicated that polyploidization events primarily led to the expansion of the plant CAMTA family, with overall similar but differentiated expansion histories in different species, and that Selaginella moellendorfii possesses the most paralogous genes being the most distinctive. More intriguingly, we found the opposite quantitative evolutionary history of terrestrial plants to that of the algal CAMTA family. Therefore, we firstly analyzed the molecular properties of the CAMTA transcription factor family in foxtail millet and explored its molecular evolutionary trajectory, and constructed a flow of evolutionary trajectories from monospecies transition to the plant kingdom for reference.

关键词： Ca2+ binding protein   Ca2+ signature   EF‐hand motif   MD simulation  

摘要： The environment surrounding plants is far from stable, compelling the plant to perceive and adapt to numerous biotic and abiotic constraints, including insect attacks. The perception of a feeding insect typically entails the identification of herbivore-associated molecular patterns causing a sequential increase in cytosolic Ca levels. Calmodulin-like proteins (CMLs) are Ca sensor proteins with conserved EF-hands, which decode Ca signals to generate a stress-specific response. Although few of the CMLs have been investigated and their role in plant defence has been deciphered in model plants, the role of CMLs as a Ca sensor protein and their interaction mechanism with calcium and downstream targets remains poorly understood in leguminous crop plants. Herein, we demonstrated the presence of a Ca signature in Glycine max during herbivory. Gene expression analysis of GmCML77 (a member of the CML family) indicated its upregulation during S. litura infestation. Phylogenetic analysis and in silico studies predicted GmCML77 as a Ca binding protein. Employing computational modelling and MD simulations, we showed that GmCML77 has predominantly α-helical conformation with 3 functional Ca binding loops. Also, Ca binding initiates an expansion of tertiary structure, leading to the exposure of hydrophobic residues that may be implicated in its interaction with target proteins. Moreover, gel shift assay and CD spectroscopy results confirmed the Ca binding ability of GmCML77. Our study demonstrated that GmCML77 is a functional calcium-binding protein, which exhibits conformational changes on Ca binding and acts as a Ca sensor during insect infestation.

关键词： GABA   Calcium   GABA transporter 3   Calmodulin   cAMP response element binding protein   Lipid rafts  

摘要： Emerging evidence underscores the crucial role of compartmentalized Ca2* and GABA signaling in the development and progression of gliomas. Our findings reveal that low GAT3 expression and high PMCA4 levels are strongly associated with poor survival outcomes in glioma patients, suggesting their involvement in tumor progression. Using C6 glioma model, we uncovered a dynamic interaction between GAT3 and PMCA4 within lipid raft microdomains, which plays a key role in fine-tuning of localized Ca2+ dynamics in response to GABA stimulation. Knockdown of PMCA4 increased resting Ca2+concentration and enhanced Ca2+ accumulation in lipid rafts following 3-min pulse GABA stimulation, significantly impairing glioma cell migration and invasion. Interestingly, the expression of Ca2+ chelator parvalbumin in rafts abolished both baseline and GABA-stimulated Ca2+ rises, effectively restoring the migratory and invasive potential of tumor cells. We further demonstrated that GAT3 interacted with calmodulin, a pivotal regulator of PMCA4, and this interaction was decreased following 24 h GABA treatment. Long-term GABA stimulation also disrupted PMCA4/GAT3 complex, overloaded lipid rafts with Ca2+ and decreased glioma invasiveness in the presence of PMCA4. In these conditions, we observed GAT3-and Ca2+/calmodulin-dependent protein kinase II-dependent CREB phosphorylation at Ser133, which was controlled by Ca2+ events in lipid rafts and required to maintain glioma invasiveness. Our study uncovers a previously unrecognized GAT3-dependent mechanism of Ca2+compartmentalization in membrane micro-domains, shedding new light on its potential role in tumor behavior. Understanding these local Ca2* signaling partnerships will offer valuable insights into gliomagenesis and could lead to the development of novel therapeutic strategies for glioma treatment.

关键词： Diseases  

摘要： The tumor suppressor p53 maintains genome stability in the setting of cellular stress and is frequently mutated in cancer. The stability of p53 is regulated by its interaction with the oncoprotein MDM2, a ubiquitin E3 ligase. Recently, nuclear phosphoinositides were reported to bind and stabilize p53. Here, we report that genotoxic stress induces the type I phosphatidylinositol phosphate kinase (PIPKIα) and its product phosphatidylinositol 4,5-bisphosphate (PIP2) to bind and regulate the stability and function of MDM2. Following genotoxic stress, nuclear PIPKIα binds to MDM2 to generate a complex of MDM2 and PIP2. PIP2 binding to MDM2 differentially regulates the recruitment of the small heat shock proteins (sHSPs) αB-crystallin (αBC) and HSP27 to the MDM2-PIP2 complex, acting as an on-off switch that regulates MDM2 stability, ubiquitination activity and interaction with p53. Our results demonstrate an unexpected role for nuclear phosphoinositides conferring specificity to the MDM2-PIP2-sHSPs association. Notably, the differential engagement of αBC and HSP27 reveals that sHSPs are not merely passive chaperones but play active, selective roles in fine-tuning MDM2 function and the MDM2-p53 nexus. These findings provide a previously unrecognized molecular framework for targeting this pathway in cancer. © 2025 The Authors

关键词： NOS   nitric oxide synthase   CaM   calmodulin   nNOS   neuronal NOS   iNOS   inducible NOS   wt   wild-type   oxyFMN   bi-domain oxygenase/FMN construct of NOS   ET   electron transfer   IET   interdomain electron transfer   XL MS   cross-linking mass spectrometry   qXL MS   quantitative cross-linking mass spectrometry   PRM   parallel reaction monitoring   DDA   data-dependent analysis   PASEF   parallel accumulation-serial fragmentation   HDX-MS   hydrogen-deuterium exchange mass spectrometry   DSBU   disuccinimidyl dibutyric urea   BSA   bovine serum albumin   FDR   false discovery rate   MSA   multiple sequence alignment   pTM   predicted template modelling   pLDDT   predicted local distance difference test   PCA   principal component analysis  

摘要： Mammalian nitric oxide synthases (NOSs) are flavo-hemoproteins that rely on dynamic interdomain interactions for activity. Calmodulin (CaM) facilitates specific, interdomain FMN–heme interactions that enable inter-subunit FMN–heme electron transfer essential for nitric oxide biosynthesis. Our quantitative cross-linking mass spectrometry (qXL MS) results demonstrate that the abundance of inter-subunit cross-links correlates with CaM-induced formation of the docked FMN/heme complex in neuronal NOS [Jiang et al., Biochemistry , 2024, 63, 1395–1411]. Here, we extend this methodology to the human inducible NOS (iNOS) isoform, comparing wild-type (wt) and E546N mutant oxygenase/FMN (oxyFMN) constructs. Using parallel reaction monitoring−based qXL MS, we assessed mutation-induced changes in interdomain dynamics. The E546N mutation substantially reduced abundance of the inter-subunit cross-links between the FMN and heme domains. Cross-links between CaM and iNOS domains were also altered by the mutation, indicating that the changes at the FMN–heme docking interface propagate allosterically throughout the iNOS−CaM complex. Although standard AlphaFold2 structural modeling yielded similar docked architectures for wt and mutant, cross-link-guided AlphaLink2 modeling revealed distinct structural differences. AlphaFold2 subsampling further predicted alternative conformations and their relative populations; consistent with qXL MS data, E546N exhibited a broader distribution of predicted conformations, with a higher undocked state population than wt. Importantly, 90% of cross-links were consistent with an ensemble of representative conformations derived from AlphaFold2 subsampling and cross-link−guided AlphaLink2 modeling, capturing both docked and undocked states alongside multiple orientations. This integrated qXL MS and AlphaFold2 subsampling strategy provides a quantitative framework for mapping mutation-induced alterations in functional dynamics of multidomain proteins.