Cyclin-dependent kinases 4 and 6 (CDK4/6) are essential drivers of cell cycle progression and have been validated as important therapeutic targets in oncology. In this study, we report the design, synthesis, and preclinical validation of a series of novel radiotracers ([68Ga]Ga-PY01-[68Ga]Ga-PY08) based on the pharmacophore of CDK4/6 inhibitor ribociclib. Flexible linkers and functionalized amino acids were incorporated to optimize pharmacokinetic and targeting properties. Among eight radiotracers, [68Ga]Ga-PY03 showed superior pharmacological and pharmacokinetic properties, including high stability, strong CDK4/6 binding affinity and low nonspecific uptake. Micro-PET/CT imaging demonstrated its capability to detect CDK4/6-overexpressed tumors and dynamically monitor CDK4/6 expression post-therapeutic. Importantly, [68Ga]Ga-PY03 also enabled quantitative assessment of CDK4/6 target occupancy, providing a potential tool for therapy response evaluation. In summary, these findings demonstrate the potential of [68Ga]Ga-PY03 as a PET radiotracer to monitor the CDK4/6 expression in tumors.

PURPOSE:CD36, a transmembrane glycoprotein, which is involved in various cellular functions, including lipid metabolism, inflammation, and tumor initiation and progression, has been considered as an important therapeutic target of tumors. However, the lack of effective imaging methods for non-invasive monitoring of CD36 expression and assessing the efficacy of CD36-targeted therapies limits the clinical application of CD36-targeted therapeutic drugs. To address this issue, we designed and synthesized two novel CD36-targeted radiotracers, and evaluated their biological properties and imaging performance, in order to select the more effective candidate for potential application in monitoring the CD36 expression and assessing the therapeutic efficacy. METHODS:The chelator NOTA was conjugated to both linear and cyclic peptides via a bioconjugation approach, yielding the linear peptide conjugate ZL01 and the cyclic peptide conjugate ZL02. Their structures were confirmed using HRMS. The solutions of peptide conjugates (ZL01 and ZL02) were mixed with a [68Ga]GaCl₃ solution to obtain the radiotracers, and the radiochemical purity of both radiotracers was determined by radio-HPLC. Non-radioactive [natGa]Ga-ZL01 and [natGa]Ga-ZL02 were used to confirm [68Ga]Ga-ZL01 and [68Ga]Ga-ZL02. The radiochemical and biological properties were evaluated, including in vitro stability, hydrophilicity, binding affinity, pharmacokinetics, micro PET/CT imaging, and biodistribution. RESULTS:[68Ga]Ga-ZL01 and [68Ga]Ga-ZL02 were obtained with radiochemical purity over 95 %. Both radiotracers demonstrated hydrophilic character, and good stability in PBS and human serum. The blood clearance of [68Ga]Ga-ZL01 and [68Ga]Ga-ZL02 (half-life) was measured at 17.8 min and 21.6 min, respectively. [natGa]Ga-ZL01 and [natGa]Ga-ZL02 exhibited high binding affinities to U87MG cells, with inhibition constant (Ki) of 1.59 ± 0.35 nM for [natGa]Ga-ZL01 and 1.12 ± 0.44 nM for [natGa]Ga-ZL02, respectively. Micro PET/CT imaging and biodistribution study revealed [68Ga]Ga-ZL02 had superior tumor-to-background ratio and prolonged tumor retention, highlighting its potential as a promising candidate for clinical translation. CONCLUSIONS:In this study, two CD36-targeted radiotracers ([68Ga]Ga-ZL01 and [68Ga]Ga-ZL02) were developed and evaluated. 68Ga-labeled cyclic peptide [68Ga]Ga-ZL02 demonstrated superior tumor-to-background ratio and prolonged tumor retention time, making it a promising radiotracer for monitoring the CD36 expression and assessing the therapeutical efficacy.

β-secretase 1 (BACE1), known for its role in amyloid-β production associated with Alzheimer’s disease (AD), has also been suggested to be elevated in patients with Type 2 diabetes mellitus (T2DM). Notably, BACE1 could cleave the insulin receptor (InsR), leading to reduced InsR levels, which may impair insulin signaling and contribute to insulin resistance. Presently, we observed decreased InsR levels and impaired glucose disposal in the livers of mice with systemic overexpression of BACE1 (HUBC mice). This suggests that elevated BACE1 could contribute to insulin resistance by shedding membrane InsR. Additionally, mice fed a high-fat diet (HFD), a well-established model of T2DM, displayed increased BACE1 levels and decreased InsR. To further investigate whether inhibiting BACE1 could enhance insulin sensitivity and alleviate symptoms of diabetes, we treated HFD mice with the BACE1 inhibitor Elenbecestat. Remarkably, the administration of Elenbecestat restored InsR levels and improved their downstream signaling pathways, leading to increased insulin sensitivity and enhanced glucose tolerance. In summary, our findings suggest that inhibiting BACE1 can restore InsR expression and improve insulin-signaling sensitivity, ultimately resulting in enhanced diabetic phenotypes.

Microplastics (MPs), plastic particles smaller than 5 mm, have garnered increasing attention due to their pervasive presence in the environment and potential health risks. While their accumulation in various organs, including the liver, kidneys, and intestines, is well-documented, the specific mechanisms they affect kidney development remain unclear. This study investigated the nephrotoxic effects of 1 μm polystyrene microplastics (PS-MPs) using a 3D kidney organoid model derived from human pluripotent stem cells, focusing on the molecular pathways involved. Kidney organoids were exposed to PS-MPs at concentrations ranging from 1.25 to 10 μg/mL for 24 h. The results demonstrated significant reductions in organoid size and nephron-specific markers, including impaired formation of proximal and distal tubules. Furthermore, enhanced autophagy and apoptosis were observed in nephron progenitor cells (NPCs), as evidenced by a 3.5-fold increase in LC3-II expression and a 1.5-fold increase in cleaved caspase-3 levels. Transcriptomic analysis identified DNA damage-inducible transcript 4 (DDIT4) as a key mediator, linking PS-MP exposure to the inhibition of mTOR signaling. Notably, silencing DDIT4 alleviated PS-MP-induced autophagy and apoptosis, highlighting its crucial role in microplastic-induced nephrotoxicity. These findings provide novel insights into the molecular pathways underlying microplastic-induced toxicity and emphasize the need for further research to explore the developmental impacts and long-term health consequences of microplastic exposure.

High mobility group box 1 (HMGB1) is highly expressed in various malignancies and has been considered as a promising molecular target for cancer diagnosis and therapy. In this study, two novel peptide-based radiotracers targeting HMGB1 were synthesized, each achieving high radiochemical purity (>95%). Of these tracers, [68Ga]Ga-HM possessed superior characteristics with a high binding affinity for the PC3 cell line (Kd = 46.11 ± 0.97 nM). Micro-PET imaging and biodistribution investigation in PC3 tumor-bearing mice revealed significant tumor uptake (SUVmax = 1.52 ± 0.10) and an excellent tumor-to-muscle ratio (6.96 ± 0.64) at 30 min post-injection. Moreover, robust imaging performance and favorable biodistribution profiles were consistently observed for [68Ga]Ga-HM across multiple distinct HMGB1-positive tumor models. These findings demonstrate that [68Ga]Ga-HM could serve as a diagnostic tracer to identify tumors with high HMGB1 expression, enabling personalized cancer treatment through the selection and monitoring of targeted therapeutic approaches.

Microplastics (MPs) have emerged as a pervasive environmental pollutant of global concern. Their detection within the human placenta and fetal organs has prompted apprehension regarding the potential hazards of MPs during early organogenesis. The kidney, a vital multifunctional organ, is susceptible to damage from MPs in adulthood. However, the precise adverse effects of MP exposure on human nephrogenesis remain ambiguous due to the absence of a suitable model. Here, we explore the potential impact of MPs on early kidney development utilizing human kidney organoids in vitro. Human kidney organoids were subjected to polystyrene-MPs (PS-MPs, 1 μm) during the nephron progenitor cell (NPC) stage, a critical phase in early kidney development and patterning. We delineate the effects of PS-MPs on various stages of nephrogenesis, including NPC, renal vesicle, and comma-shaped body, through sequential examination of kidney organoids. PS-MPs were observed to adhere to the surface of cells during the NPC stage and accumulate within glomerulus-like structures within kidney organoids. Moreover, both short- and long-term exposure to PS-MPs resulted in diminished organoid size and aberrant nephron structure. PS-MP exposure heightened reactive oxygen species (ROS) production, leading to NPC apoptosis during early kidney development. Increased apoptosis, diminished cell viability, and NPC reduction likely contribute to the observed organoid size reduction under PS-MP treatment. Transcriptomic analysis at both NPC and endpoint stages revealed downregulation of Notch signaling, resulting in compromised proximal and distal tubular structures, thereby disrupting normal nephron patterning following PS-MP exposure. Our findings highlight the significant disruptive impact of PS-MPs on human kidney development, offering new insights into the mechanisms underlying PS-MP-induced nephron toxicity.

Neural tube defects (NTDs) represent a developmental disorder of the nervous system that can lead to significant disability in children and impose substantial social burdens. Valproic acid (VPA), a widely prescribed first-line antiepileptic drug for epilepsy and various neurological conditions, has been associated with a 4-fold increase in the risk of NTDs when used during pregnancy. Consequently, urgent efforts are required to identify innovative prevention and treatment approaches for VPA-induced NTDs. Studies have demonstrated that the disruption in the delicate balance between cell proliferation and apoptosis is a crucial factor contributing to NTDs induced by VPA. Encouragingly, our current data reveal that melatonin (MT) significantly inhibits apoptosis while promoting the restoration of neuroepithelial cell proliferation impaired by VPA. Moreover, further investigations demonstrate that MT substantially reduces the incidence of neural tube malformations resulted from VPA exposure, primarily by suppressing apoptosis through the modulation of intracellular reactive oxygen species levels. In addition, the Src/PI3K/ERK signaling pathway appears to play a pivotal role in VPA-induced NTDs, with significant inhibition observed in the affected samples. Notably, MT treatment successfully reinstates Src/PI3K/ERK signaling, thereby offering a potential underlying mechanism for the protective effects of MT against VPA-induced NTDs. In summary, our current study substantiates the considerable protective potential of MT in mitigating VPA-triggered NTDs, thereby offering valuable strategies for the clinical management of VPA-related birth defects.

ABSTRACT:Objective: To evaluate the effectiveness of a general education course titled "The Basis of Radiation Protection" in building and strengthening undergraduate awareness of radiation safety and cultivating innovative individuals with reasonable knowledge structures and strong practical abilities. Methods: All students from 2021 to 2022 enrolled in the core general education course "The Basis of Radiation Protection" at Shandong University of China were invited to participate. A questionnaire survey was conducted to determine changes in the students' basic cognition of radiation safety and scientific protection before and after the course. Results: The survey indicated that the cognitive level of radiation science protection had significantly improved through course completion. The Liszt quantification score range increased from 3.45 to 4.77 to 4.81 to 4.98 (p < 0.001). Further analysis revealed that different professional backgrounds significantly affected students' understanding of radiation safety protection; medical students were superior to electrical engineering students in their knowledge of ionizing radiation before the course (p < 0.001). However, after course completion, the understanding of students from both majors regarding radiation safety had relatively improved, and no significant difference was detected (p > 0.05). Feedback on the course showed that the awareness of "daily radiation protection" had significantly improved (96.8%), pseudoscience and pseudo-information could be correctively identified (93.6%), "nuclear power"-related fears had been dispelled (95.7%), and the concept of "cherishing life" had been effectively established (91.5%). Conclusion: The course effectively improved the awareness of radiation safety, strengthened the knowledge system, and provided a new way to cultivate innovative talent with reasonable knowledge structures.

This study evaluated the effects of chronic intermittent hypoxia (CIH) at different times on the mitochondria of mouse hearts and H9C2 cardiomyocytes to determine the role of the cannabinoid receptor 1 (CB1R)/adenosine 5'-monophosphate–activated protein kinase (AMPK)/peroxisome proliferator-activated receptor-γ coactivator-1α (PGC-1α) signaling pathway. Animal and cellular CIH models were prepared in an intermittent hypoxia chamber at different times. The cardiac function of mice was determined, and heart tissue and ultrastructural changes were observed. Apoptosis, reactive oxygen species (ROS), and mitochondrial membrane potential were detected, and MitoTracker™ staining was performed to observe cardiomyocyte mitochondria. Western blot, immunohistochemistry, and cellular immunofluorescence were also performed. In the short-term CIH group, increases in mouse ejection fraction (EF) and heart rate (HR); mitochondrial division; ROS and mitochondrial membrane potential; and the expression levels of CB1R, AMPK, and PGC-1α were observed in vivo and in vitro. In the long-term CIH group, the EF and HR increased, the myocardial injury and mitochondrial damage were more severe, mitochondrial synthesis decreased, the apoptosis percentage and ROS increased, mitochondrial fragmentation increased, membrane potential decreased, CB1R expression increased, and AMPK and PGC-1α expression levels decreased. Targeted blocking of CB1R can increase AMPK and PGC-1α, reduce damage attributed to long-term CIH in mouse hearts and H9C2 cells, and promote mitochondrial synthesis. Short-term CIH can directly activate the AMPK/PGC-1α pathway, promote mitochondrial synthesis in cardiomyocytes, and protect cardiac structure and function. Long-term CIH can increase CB1R expression and inhibit the AMPK/PGC-1α pathway, resulting in structural damage, the disturbance of myocardial mitochondria synthesis, and further alterations in the cardiac structure. After targeted blocking of CB1R, levels of AMPK and PGC-1α increased, alleviating damage to the heart and cardiomyocytes caused by long-term CIH.

Objective:To assess the impact of the general education course entitled " The Basics of Radiation Protection" on enhancing undergraduates′ awareness of radiation safety, a questionnaire was utilized. Additionally, this study aims to propose novel strategies for improving health literacy among undergraduates, fostering the development of innovative, talented individuals with reasonable knowledge structures and strong practical abilities.Methods:From 2021 to 2022, 127 undergraduates from all colleges of Shandong University took the general course" The Basics of Radiation Protection". To compare the learning effects of the medical and non-medical students, invitations to participate in this study were extended to 101 (79.5%) students from School of Medicine and School of Electrical Engineering. A questionnaire survey was conducted to determine and compare the changes in the students’ basic cognition of radiation safety and protection before and after learning the course. The aim was to evaluate the teaching effectiveness and significance of the course.Results:From 2021 to 2022, 94 undergraduates from School of Medicine and School of Electrical Engineering accepted the invitation and voluntarily completed questionnaires twicely, accounting for 74.0% of the total number of selected students. The questionnaire survey showed that through course learning, the cognitive level of radiation protection among college students had significantly improved. The Likert quantification scores befroe taking the course were 3.59±1.35, 3.81±1.17, 3.45±1.16, 4.11±1.13, 3.62±1.05, 3.98±1.24 and after completing the course 4.94±0.32, 4.87±0.32, 4.96±0.21, 4.98±0.15, 4.81±0.39, 4.98±0.15, respectively, with the difference being statistically significant ( t=6.55, 5.50, 9.10, 5.36, 7.60, 5.34, P< 0.05). Further analysis revealed that different professional backgrounds affected students′ understanding of radiation safety and protection: before learning this course, medical students were superior to electrical engineering students in their knowledge of ionizing radiation ( t=5.69, 4.06, 6.46, 6.27, 7.78, P < 0.001). After taking the course, the cognitive level of students from both majors had considerably improved, but there was no significant difference ( P> 0.05). Feedback on the course learning showed that the awareness of " routine radiation protection" had significantly improved (96.8%), pseudoscience and pseudoinformation could be identified (93.6%), " nuclear power" -related fears had been dispelled (95.7%), and the concept of " cherishing life" had been effectively established (91.5%). Conclusions:The general education course " The Basics of Radiation Protection" has effectively improved the students′ awareness of radiation safety, strengthened their knowledge regarding radiation safety and protection, and provided a new avenue for cultivating innovative talents on a well-structured knowledge base. The course would be worth further promoting and applying in the education of undergraduates with different backgrounds and taking different majors.

Macrophage (MΦ) with different surface marker exhibits diversity effect on the development of atherosclerosis (AS). Recently CD93high MΦ was reported tightly related with inflammatory diseases, however, its effect on the development of AS remains unclear. Here, we adoptive transferred with prepared CD93 high and low expressed (CD93hi, CD93lo) MΦ in establishing ApoE-/- mice AS model at the week of 1, 4, 8, 12, 16, respectively. At w12 and 20, pathology change and the expression of CD93, F4/80, CD31, ApoE in related vessels were detected. Pro-inflammatory cytokines and related signal pathway were also analyzed. Finally, CD93 targeting radionuclide imaging in AS model was performed with prepared 125I-anti-CD93 mAb injection. The results showed that all pathological changes in diseased vessels showed much severe in w20 (the time that AS model was established successfully) than w12. In contrast to group with CD93lo MΦ adoptive transferred, much serious inflammatory infiltration, increased pro-inflammatory factors and elevated CD93, F4/80, CD31 expression in clamped carotid artery were detected, and also higher Tumor necrosis factor-α (TNF-α) and Interleukin-1β (IL-1β) in serum were measured in ApoE-/- model with CD93hi MΦ adoptive transferred in w20. Furthermore, obviously high expressed p-STAT3 and p-NF-κB were detected in CD93hi MΦ, compared with CD93lo MΦ. Ex vivo phosphor-autoradiography showed higher radioactivity accumulation in diseased vessels in ApoE-/- model with CD93hi MΦ adoptive transferred after 125I-anti-CD93 mAb injection, compared with CD93lo MΦ adopted group in w20. In conclusion, CD93hi MΦ subset promoted the development of AS and aggravated the disease severity mainly through p-STAT3/p-NF-κB pathways. CD93 on MΦ may be as a novel target for AS modulation.

A new strategy for the early diagnosis of triple-negative breast cancer (TNBC) is urgently needed however specific targets are lacking. EphA2 has been reported to be over-expressed in a variety of tumors, including TNBC, and is closely related to tumor progression. In this study, we designed a novel peptide, SD01, and tested its potential in the diagnosis of TNBC. FITC-SD01 and FITC-YSA were prepared and found to bind to the 4T1 TNBC cell line, the former showing greater affinity. 125I-SD01 and 125I-YSA were obtained with high radiochemical yield and radiochemical purity, and both showed a high binding affinity to 4T1 cells, with a higher Bmax in 125I-SD01. Whole-body phosphoautoradiography showed clearer imaging of tumors in the group of 125I-SD01 than in 125I-YSA. Biodistribution demonstrated higher tumor accumulation in the 125I-SD01 group. In group of 125I-SD01, the tumor to the opposite muscle tissue (T/NT) ratio was 5.998 ± 0.37, in contrast to 4.69 ± 0.18 in 125I-YSA group. Our results indicated that 125I-SD01 could selectively and specifically target 4T1 cells in vitro and in vivo, and showed higher binding affinity and better imaging compared to 125I-YSA. Therefore, SD01 was deemed to be a novel peptide with more favorable properties in terms of targeting EphA2.

Noninvasive imaging atherosclerotic (AS) plaque is of great importance for early diagnosis. Recently, CD93 in MΦ was linked to atherosclerosis development. Herein, we have investigated whether CD93 in MΦ is a potential novel target for atherosclerotic plaque imaging. CD93hi and CD93lo MΦ were prepared with or without LPS stimulation, before biological activity was evaluated. A rat AS model was produced with left carotid artery clamped. Whole-body/ex vivo phosphor autoradiography of the artery and biodistribution were investigated after incorporation of 3 H-2-DG into CD93hi and CD93lo MΦ or after 125 I-α-CD93 (125 I-anti-CD93mAb) injection. The plaque tissue was subjected to CD93/CD68 immunofluorescence/immunohistochemistry staining. CD93hi and CD93lo MΦ cells were successfully prepared without significant effect on bioactivity after incorporative labelled with 3 H-2-DG. The AS model was successfully established. Biodistribution studies showed that adoptive transfer of 3 H-2-DG-CD93hi MΦ or 125 I- α-CD93 injection resulted in accumulation of radioactivity within the atherosclerotic plaque in the clamped left carotid artery. T/NT (target/non-target, left/right carotid artery) ratio was higher in the 3 H-2-DG-CD93hi MΦ adoptive transfer group than in the 3 H-2-DG-CD93lo MΦ group (p < .05). Plaque radioactivity in the 125 I-α-CD93 injection group was significantly higher than in the 125 I-IgG control group (p < .01). The higher radioactivity accumulated in the clamped left carotid artery was confirmed by phosphor autoradiography. More importantly, CD93/CD68 double-positive MΦ accumulated at the atherosclerotic plaque in 3 H-2-DG-CD93hi MΦ adoptive transfer group, which correlated with plaque radioactivity (r = .99, p < .01). In summary, both adoptive-transferred 3 H-2-DG-labelled CD93hi MΦ and 125 I-α-CD93 injection specifically targeted CD93 in atherosclerotic plaque. CD93 is a potential target in atherosclerotic plaque imaging.

探索基础医学综合实验教学新模式,提高学生创新能力.构建了细胞生物、组织病理、放射性示踪等多学科交叉融合的综合实验教学体系.以肿瘤形成全过程为时间轴,设计多维度、实时观测的实验教学单元;建立了多元化、综合考核方式,评价实验教学质量.基于肿瘤形成全过程的综合创新实验课程有利于学生实践能力和创新能力的提升.

Haze problem is an important factor threatening human health. PM2.5 is the main culprit haze. 1-Nitropyrene (1-NP) is the main nitrated polycyclic aromatic hydrocarbon, the toxic component of PM2.5 particles. The effects of 1-NP on various organs and reproductive health have been extensively and deeply studied, but the effects of 1-NP on embryo implantation and endometrial receptivity remain to be determined. The purpose of this study was to investigate the adverse effects of 1-NP on mouse embryo implantation and human endometrial receptivity. In early pregnancy, CD1 mice were given 2 mg/kg 1-NP by oral gavage, which resulted in a decreased embryo implantation number on day 5, inhibited leukemic inhibitory factor (LIF)/STAT3 pathway, decreased expression of estrogen receptor and progesterone receptor, and disrupted regulation of uterine cell proliferation. In addition, in a human in vitro implantation model, 1-NP was found to significantly inhibit the adhesion rate between trophoblast spheroids and endometrial epithelial cells, possibly by inhibiting the expression of receptivity molecules in Ishikawa cells. Promoting reactive oxygen species (ROS) production may be an additional mechanism by which it inhibits trophoblast spheroid adhesion. In this study, we used an in vivo mouse pregnancy model and an in vitro human embryo implantation model to demonstrate that 1-NP can impair endometrial receptivity and compromise embryo implantation.

In this study, GFP-tagged TNBC 4T1 cells with down-regulated TLR5 expression (TLR5− 4T1) and normal TLR5 expression (TLR5+ 4T1) were constructed, respectively. RT-PCR and Western blot studies showed that down-regulation of TLR5 obviously increased the expression of VEGFR in 4T1 cells. Highly stable radio-probes 125I-anti-TLR5 mAb/125I-VEGF/125I-IgG were obtained with labeling rates over 85% and radiochemical purities above 90%. Among these three probes, 125I−anti−TLR5 mAb and 125I-VEGF were used for specifically imaging TNBC, while 125I-IgG was used for comparison. Whole-body phosphorus autoradiography showed clear imaging at 48 h after injection of 125I-anti-TLR5 mAb and 125I-VEGF also provided clear imaging at 24 h. Biodistribution study demonstrated a higher tumor uptake of 125I-anti-TLR5 mAb in TLR5+ group compared with that in TLR5− group (P < 0.05), whereas tumor uptake of 125I-VEGF in TLR5+ group was lower than that in the TLR5− group (P < 0.05). Immunohistochemical staining suggested that the expression of TLR5 was lower, whereas the expression of VEGFR, CD31, and MVD (microvessel density) was higher in TLR5− tumor-bearing mice. In summary, the down-regulation of TLR5 in TNBC promoted the VEGFR expression and angiogenesis, resulting in the proliferation of TNBC cells. TLR5/VEGF might be a better indicator for monitoring the development of TNBC.

Lung cancer is the most common malignancy and leads to around one-quarter of all cancer deaths. Great advances have been achieved in the treatment of lung cancer with novel anticancer agents and improved technology. However, morbidity and mortality rates remain extremely high, calling for an urgent need to develop novel anti–lung cancer agents. 1,2,3-Triazole could be readily interact with diverse enzymes and receptors in organisms through weak interaction. 1,2,3-Triazole can not only be acted as a linker to tether different pharmacophores but also serve as a pharmacophore. This review aims to summarize the recent advances in 1,2,3-triazole–containing compounds with anti–lung cancer potential, and their structure–activity relationship (SAR) together with mechanisms of action is also discussed to pave the way for the further rational development of novel anti–lung cancer candidates.

CD93分子作为一种新型新生血管生成激活剂,有可能成为肿瘤监测的重要分子靶点,因此制备125 I标记抗CD93单抗(125 I-anti-CD93 mAb),研究其在荷瘤鼠体内生物学分布及磷屏显像特点,探讨其对A549肺癌靶向性及CD93阳性肿瘤监测的可行性.本文制备125 I-anti-CD93 mAb及125 I-IgG并进行鉴定;建立A549肺癌荷瘤小鼠模型,经尾静脉注射125 I标记抗CD93单抗,并设125 I-IgG为对照组,注药后不同时间进行生物学分布及磷屏放射自显影;肿瘤组织行H E染色及CD93免疫组织化学染色;采用统计软件对定量数据(x-+s)进行统计学处理,组间数据用t检验,P<0.05为差异有统计学意义.结果表明,125 I-anti-CD93 mAb及125 I-IgG标记率、放射性比活度、放化纯度分别为91.37％ 和90.24％,1096.44 MBq/mg和1082.88 MBq/mg,96.49％ 和94.82％,放置72 h后两标记物的稳定性仍>90％,两者间无统计学差异(生理盐水组P=0.93,t=0.09,血清组P=0.13,t=1.90).荷瘤鼠生物学分布结果显示125 I-anti-CD93 mAb主要经肝肾代谢,注射后48 h时肿瘤组织放射性摄取率为(6.42±0.71)％ID/g,T/NT(瘤/对侧肌肉)摄取比值为4.45±0.86,显著高于对照组125I-IgG组(2.45±0.33/1.71±0.24),P<0.05,t=3.07和P<0.01,t=5.10.动态全身磷屏自显影结果可见,24 h两组小鼠轮廓清楚,48 h时实验组肿瘤显像最清晰,肿瘤放射性摄取比(肿瘤/对侧肌肉)为3.34±0.18,明显高于对照组(1.62±0.19),P<0.01,t=6.52.免疫组化染色结果证实肿瘤高表达CD93.125 I-anti-CD93 mAb制备简便,对A549肺癌组织靶向性好,有望成为CD93阳性肿瘤监测的新型分子探针.

Non-invasively monitoring allogeneic graft rejection with a specific marker is of great importance for prognosis of patients. Recently, data revealed that IL-27Rα was up-regulated in alloreactive CD4+ T cells and participated in inflammatory diseases. Here, we evaluated whether IL-27Rα could be used in monitoring allogeneic graft rejection both in vitro and in vivo. Allogeneic (C57BL/6 donor to BALB/c recipient) and syngeneic (BALB/c both as donor and recipient) skin grafted mouse models were established. The expression of IL-27Rα in grafts was detected. The radio-probe, 125I-anti-IL-27Rα mAb, was prepared. Dynamic whole-body phosphor-autoradiography, ex vivo biodistribution and immunofluorescence staining were performed. The results showed that the highest expression of IL-27Rα was detected in allogeneic grafts on day 10 post transplantation (top period of allorejection). 125I-anti-IL-27Rα mAb was successfully prepared with higher specificity and affinity. Whole-body phosphor-autoradiography showed higher radioactivity accumulation in allogeneic grafts than syngeneic grafts on day 10. The uptake of 125I-anti-IL-27Rα mAb in allogeneic grafts could be almost totally blocked by pre-injection with excess unlabeled anti-IL-27Rα mAb. Interestingly, we found that 125I-anti-IL-27Rα mAb accumulated in allogeneic grafts, along with weaker inflammation earlier on day 6. The high uptake of 125I-anti-IL-27Rα mAb was correlated with the higher infiltrated IL-27Rα positive cells (CD3+/CD68+) in allogeneic grafts. In conclusion, IL-27Rα may be a novel molecular imaging marker to predict allorejection.

Recently, emerging evidence strongly suggested that the activation of interleukin-27 Receptor α (IL-27Rα) could modulate different inflammatory diseases. However, whether IL-27Rα affects allotransplantation rejection is not fully understood. Here, we investigated the role of IL-27Rα on allorejection both in vivo and in vitro. The skin allotransplantation mice models were established, and the dynamic IL-27Rα/IL-27 expression was detected, and IL-27Rα+ spleen cells adoptive transfer was performed. STAT1/3/5 phosphorylation, proliferation and apoptosis were investigated in mixed lymphocyte reaction (MLR) with recombinant IL-27 (rIL-27) stimulation. Finally, IFN-γ/ IL-10 in graft/serum from model mice was detected. Results showed higher IL-27Rα/IL-27 expression in allografted group compared that syngrafted group on day 10 (top point of allorejection). IL-27Rα+ spleen cells accelerated allograft rejection in vivo. rIL-27 significantly promoted proliferation, inhibited apoptosis and increased STAT1/3/5 phosphorylation of alloreactive splenocytes, and these effects of rIL-27 could be almost totally blocked by JAK/ STAT inhibitor and anti-IL-27 p28 Ab. Finally, higher IL-27Rα+ IFN-γ+ cells and lower IL-27Rα+ IL-10+ cells within allografts, and high IFN-γ/low IL-10 in serum of allorejecting mice were detected. In conclusion, these data suggested that IL-27Rα+ cells apparently promoted allograft rejection through enhancing alloreactive proliferation, inhibiting apoptosis and up-regulating IFN-γ via enhancing STAT pathway. Blocking IL-27 pathway may favour to prevent allorejection, and IL-27Rα may be as a high selective molecule for targeting diagnosis and therapy for allotransplantation rejection.