[Objective] To compare the characteristics of platelet-rich plasma derived exosomes (PRP-Exos) under different activation conditions and their differential effects on the proliferation capacit of human microvascular endothelial cells (HMEC-1) and human skin fibroblasts (BJ). [Methods] Ten healthy volunteers were recruited, and 10 mL of venous blood anticoagulated with EDTA-K2 was collected. PRP-Exos were prepared and extracted by the secondary centrifugation method. Transmission electron microscopy (TEM), nanoparticle tracking analysis (NTA), and Western blotting (WB) detection techniques were used to compare the morphological characteristics, particle size distribution, concentration, and the expression of surface marker proteins of PRP-Exos. Through in vitro cell culture, the differences in morphological changes and proliferation abilities of HMEC-1 and BJ cells induced by PRP-Exos in different activator groups were compared. [Results] 1) TEM observation showed that the morphologies of PRP-Exos in different activator groups were different. 2) NTA analysis showed that compared with the particle size of (109.60±2.71) nm in the normal saline group, the particle sizes of the thrombin group [(104.93±1.55) nm] and the mixed agent group [(103.83±1.58) nm] were relatively smaller, while the particle size of the calcium gluconate group [(113.57±4.70) nm] was larger and its particle size distribution range was wider. There were statistically significant differences among different groups (F=7.019, P<0.05). The concentration of exosomes obtained in the group with the mixture of calcium gluconate and thrombin was the highest [(4.87±0.63)×10⁶ particles/mL, P<0.001]. Both the thrombin group [(2.75±0.13)×10⁶ particles/mL, P < 0.01] and the calcium gluconate group [(3.82±0.06)×10⁶ particles/mL, P<0.001] obtained higher concentrations of exosomes compared with the normal saline group. The concentration in the calcium gluconate group was slightly higher than that in the thrombin group, and there was a significant difference between the two groups (P<0.05). 3) WB analysis showed that CD41, CD81, and TSG101 were expressed on the surface of PRP-Exos, and the protein signal intensities of CD41 and TSG101 in the group with the mixture of thrombin and calcium gluconate were the strongest (P<0.001). 4) The results of in vitro cell culture showed that PRP-Exos in the group with the mixture of thrombin and calcium gluconate could significantly promote the proliferation of HMEC-1 and BJ cells (P<0.05). [Conclusion] The PRP treated with the thrombin and calcium gluconate mixture group yielded the highest concentration of exosomes; under in vitro culture conditions, PRP-Exos from this group significantly promoted the proliferation of HMEC-1 and BJ cells.

BACKGROUND:Growth factor is the key effect molecule that plays a role in platelet-rich plasma in clinical treatment.There are differences in the concentration of growth factor after different activators activate platelet-rich plasma,which is an important factor affecting clinical efficacy. OBJECTIVE:To analyze the influence of different activators on the mass concentration of growth factors in platelet-rich plasma. METHODS:Totally 12 healthy volunteers were recruited to collect EDTA-K2 anticoagulant venous blood.Secondary centrifugation was used to prepare platelet-rich plasma.The difference in mass concentrations of growth factors was compared between venous blood and platelet-rich plasma.The platelet-rich plasma was mixed with four activators(normal saline,thrombin,calcium gluconate,calcium gluconate+thrombin)according to the volume ratio of 10:1,and incubated in a constant temperature water bath at 37 °C for 30 minutes.After centrifugation,the supernatant was extracted and the mass concentration of growth factor was detected.The bacterial growth in supernatant was measured by blood agar plate.Pearson correlation was used to analyze the correlation between different activators and the mass concentration of growth factor in platelet-rich plasma,and the correlation between the value of thrombocytometer and the mass concentration of growth factors in platelet-rich plasma. RESULTS AND CONCLUSION:(1)The mass concentrations of platelet-derived growth factor-BB,platelet-derived growth factor-AB,vascular endothelial growth factor,and epidermal growth factor in platelet-rich plasma were 8.7,22.2,2.3,and 2.8 times of those in venous blood,respectively(P<0.05).(2)Compared with normal saline group,the mass concentrations of platelet-derived growth factor BB,platelet-derived growth factor AB,vascular endothelial growth factor,and epidermal growth factor were increased in the thrombin group,calcium gluconate group,and calcium gluconate+thrombin group(P<0.05).The mass concentration of platelet-derived growth factor BB in the thrombin group and calcium gluconate group was higher than that in the calcium gluconate+thrombin group(P<0.05),and the mass concentration of platelet-derived growth factor AB in the thrombin group was higher than that in the calcium gluconate group and calcium gluconate+thrombin group(P<0.05).Epidermal growth factor mass concentration in the thrombin group was lower than that in the calcium gluconate group and calcium gluconate+thrombin group(P<0.05).(3)The results of blood agar plate test showed no bacterial growth in the supernatant of the four groups.(4)Pearson correlation analysis showed that the mass concentration of platelet-derived growth factor BB in platelet-rich plasma was strongly positively correlated with thrombin(r=0.683,P<0.05),and the mass concentration of vascular endothelial growth factor was strongly positively correlated with thrombin,calcium gluconate,calcium gluconate+thrombin stimulant(r=0.730,0.789,0.686,P<0.05).There was no correlation between the value of thrombocytometer and the mass concentration of four kinds of growth factors(P>0.05).(5)The results suggest that different activators have an impact on the concentration of growth factors in platelet-rich plasma.It is suggested to choose different activators to improve clinical efficacy according to different growth factor mass concentrations and treatment needs.

Radon, the only naturally occurring radioactive noble gas, is among the most common radioactive nuclides to which humans are exposed. Radon can induce various biological effects in the human body and is a risk factor for lung cancer. Circular RNAs (circRNAs) are stable, tissue-specific, and abundantly expressed in body fluids. circRNAs can regulate gene expression and play an important role in the development of cancer. In this paper, we summarized the changes in the expression and function of circRNAs, highlighting the potential mechanisms of circRNAs in radon exposure-induced cancers. Our results provided theoretical support for the use of circRNAs as a biomarker of radon exposure-induced radiation damage, and offer a theoretical basis for the early diagnosis, treatment, and prevention of radon exposure-induced diseases.

BACKGROUND: Several studies have demonstrated the occurrence of secondary tumors as a rare but significant complication of chimeric antigen receptor T (CAR-T) cell therapy, underscoring the need for a detailed investigation. Given the limited variety of secondary tumor types reported to date, a comprehensive characterization of the various secondary tumors arising after CAR-T therapy is essential to understand the associated risks and to define the role of the immune microenvironment in malignant transformation. This study aims to characterize the immune microenvironment of a newly identified secondary tumor post-CAR-T therapy, to clarify its pathogenesis and potential therapeutic targets. METHODS: In this study, the bone marrow (BM) samples were collected by aspiration from the primary and secondary tumors before and after CD19 CAR-T treatment. The CD45 + BM cells were enriched with human CD45 microbeads. The CD45 + cells were then sent for 10× genomics single-cell RNA sequencing (scRNA-seq) to identify cell populations. The Cell Ranger pipeline and CellChat were used for detailed analysis. RESULTS: In this study, a rare type of secondary chronic myelomonocytic leukemia (CMML) were reported in a patient with diffuse large B-cell lymphoma (DLBCL) who had previously received CD19 CAR-T therapy. The scRNA-seq analysis revealed increased inflammatory cytokines, chemokines, and an immunosuppressive state of monocytes/macrophages, which may impair cytotoxic activity in both T and natural killer (NK) cells in secondary CMML before treatment. In contrast, their cytotoxicity was restored in secondary CMML after treatment. CONCLUSIONS This finding delineates a previously unrecognized type of secondary tumor, CMML, after CAR-T therapy and provide a framework for defining the immune microenvironment of secondary tumor occurrence after CAR-T therapy. In addition, the results provide a rationale for targeting macrophages to improve treatment strategies for CMML treatment.
Humans ; Lymphoma, Large B-Cell, Diffuse/therapy* ; Tumor Microenvironment/genetics* ; Antigens, CD19/metabolism* ; Leukemia, Myelomonocytic, Chronic/genetics* ; Immunotherapy, Adoptive/adverse effects* ; Male ; Single-Cell Analysis/methods* ; Female ; Sequence Analysis, RNA/methods* ; Receptors, Chimeric Antigen ; Middle Aged

The persistent high prevalence and poor survival outcomes of lung cancer underscore the urgent need for innovative therapeutic modalities. Here, we present a novel multifunctional delivery platform for the synergistic treatment of lung malignancies, combining in situ-triggerable photodynamic therapy (PDT) with radiotherapy. The new platform CLL was developed by loading a new reactive oxygen species (ROS)-triggerable photosensitizer, luminol-conjugated chlorin e6 (Ce6), into liposomes. CLL can be activated through the bioluminescence resonance energy transfer effect under oxidative stress, thereby producing singlet oxygen for targeted tumor treatment without external irradiation. In vitro studies showed significant cytotoxic effects of CLL in both 4T1 and A549 tumor cells. Furthermore, a PDT-radiopharmaceutical combination nanotherapy CLL-¹⁷⁷Lu was engineered by incorporating the radionuclide ¹⁷⁷Lu into CLL. CLL-¹⁷⁷Lu demonstrated synergistic antitumor effects in 4T1 and A549 tumor cells, as well as in mouse models of 4T1 breast cancer lung metastasis or A549 tumor xenografts. Mechanistically, CLL-¹⁷⁷Lu can induce singlet oxygen/ROS generation, enhance tumor cell apoptosis, and promote M1 macrophage-mediated immunotherapy. Preliminary assessments showed a favorable profile for CLL-¹⁷⁷Lu, highlighting its potential as a promising nanotherapy for cancer treatment. Additionally, CLL can serve as a versatile platform for delivering a range of therapies to achieve synergistic antitumor effects.

Accumulating evidence has demonstrated that nucleic acid-based therapies are promising for atherosclerosis. However, nearly all nucleic acid delivery systems developed for atherosclerosis necessitate injection, which results in rapid elimination and poor patient compliance. Consequently, oral delivery strategies capable of targeting atherosclerotic plaques are imperative for nucleic acid therapeutics. Herein we report the development of yeast-derived capsules (YCs) packaging an antisense oligonucleotide (AM33) targeting microRNA-33 (miR-33) for the oral treatment of atherosclerosis. YCs provide stability for AM33, preventing its premature release in the gastrointestinal tract. AM33-containing YCs, defined as YAM33, showed high transfection in macrophages, thus promoting cholesterol efflux and inhibiting foam cell formation by regulating the target genes/proteins of miR-33. Orally delivered YAM33 effectively accumulated within atherosclerotic plaques in ApoE ^-/- mice, primarily by transepithelial absorption via M cells in Peyer's patches and subsequent translocation via macrophages through the lymphatic system. Inhibition of miR-33 by oral YAM33 significantly delayed the progression of atherosclerosis. Moreover, oral treatment with YCs co-delivering AM33 and atorvastatin afforded significantly enhanced anti-atherosclerotic effects. Our findings suggest that yeast-based microcapsules represent an effective carrier for oral delivery of nucleic acids, either alone or in combination with existing drugs, offering a promising approach for precision therapy of atherosclerotic diseases.

Objective To compare the therapeutic effects of modified Shenqi Maiwei Dihuang Decoction combined with Buzhong Yiqi Pill and Buzhong Yiqi pill alone in the patients with cold and heat mixed type di-abetic foot.Methods A total of 123 patients with cold and heat mixed type diabetic foot receiving the treat-ment in this hospital from April 2022 to December 2023 were selected and divided into the modified Shenqi Mai-Dihuang Decoction and Buzhong Yiqi Pill group(combined group,60 cases)and Buzhong Yiqi Pill group(monotherapy group,63 cases).The wound healing,mean blood flow amount of foot dorsal artery,blood glu-cose and changes in vascular diameter,inflammatory factors and vascular endothelial growth factor after 2 courses of treatment were compared betweeb the 2 groups.Results The ulcer reduction rate in the combina-tion group was significantly higher than that in the monotherapy group(P=0.001).The fasting blood glu-cose and 2 h postprandial blood glucose after treatment in the combination group all were lower than those in the monotherapy group(P=0.001).The average blood flow of foot dorsal artery after treatment in combina-tion group was higher than that in the monotherapy group,and the difference was statistically significant(P=0.013).There were 34 cases of Wagner grade 3 in the combined group and 33 cases of Wagner grade 3 in the monotherapy subgroup respectively.There was statistically significant difference in the CRP level between the two subgroups(P=0.045).Conclusion Shenqi Maiwei Dihuang Decoction combined with Buzhong Yiqi Pill in treating diabetic foot ulcer is more effective than Buzhong Yiqi Pill alone,moreover the safety is high.

Objective This study aimed to explore the differences in global and local brain network topological properties between deficient pattern(DP)and excess pattern(EP)of mild vascular cognitive impairment caused by subcortical small vessel disease based on graph theory network.Methods Patients were recruited prospectively and were classified with DP and EP subtype.The global small-world topological attributes and local nodes were calculated for the comparison of DP,EP,and healthy controls(CN)using the GRETNA platform.Results The three groups all had small-world attributes,but only the patients in EP had a significantly lower small world attribute δ in the range of 0.05-0.26 than the control group(P<0.05).The node efficiency and node strength indicators of multiple brain region were able to significantly distinguish the DP group from the EP group.However,there was no positive brain region in the node efficiency of the DP patients(P>0.05),and only a few brain regions showed increased node strength efficiency(P<0.05).Conclusion The results indicate that the syndrome of DP and EP have significantly different neuroimaging phenotypes,providing a basis for further research of biological classification based on Chinese Medicine syndromes.

Chondrosarcoma(CHS)is the second most common primary bone malignancy in human.Recurrence and metasta-sis are common among patients,and the efficacies of radiotherapy and chemotherapy are limited.The development of CHS treat-ment method has stagnated in the past 30-40 years,and surgical treatment is still the dominant option.Research on the patho-genesis mechanism of CHS is still not clear,and there is no specific drug.Therefore,it is of great significance to explore the pathogenesis of CHS,so as to find new treatment methods and drug targets for clinical treatment.Some studies have shown that epigenetic factor play an important role in the pathogenesis of CHS.Epigenetics refers to changes in the expression level and function of a gene without the change in its DNA sequence,leading to a heritable phenotype.The mechanisms of epigenetics mainly include DNA methylation,non-coding RNA regulation,histone modification,and chromatin remodeling.In this review,the recent progress in the field of epigenetics in CHS is summarized,which provides a direction for further research on CHS and searching for new therapeutic targets.