[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.

ObjectiveTo investigate the differences in abnormal liver biochemical parameters in pregnant patients during the epidemic or non-epidemic period of coronavirus disease 2019 （COVID-19）. MethodsA retrospective analysis was performed for 539 pregnant women who were discharged from Department of Obstetrics， Peking University First Hospital， from October 2017 to March 2022 and had at least one abnormal liver biochemical parameter among alanine aminotransferase （ALT）， aspartate aminotransferase （AST）， gamma-glutamyl transpeptidase （GGT）， total bilirubin （TBil）， and total bile acid. The patients in the epidemic period of COVID-19 and those in the non-epidemic period of COVID-19 were compared in terms of etiology， coagulation parameters， aminotransferases， bile acid， and renal function. The independent samples t-test was used for comparison of continuous data between two groups， and the chi-square test was used for comparison of categorical data between two groups. ResultsAmong the patients discharged from Department of Obstetrics during the non-epidemic period of COVID-19， 262 had abnormal liver biochemical parameters， accounting for 1.46%， while 277 patients had abnormal liver biochemical parameters during the epidemic period of COVID-19， accounting for 1.73% among the patients discharged from Department of Obstetrics during the same period of time， and there was a significant difference between these two groups （χ²=3.947， P=0.047）. The etiological analysis of the patients with abnormal liver biochemical parameters during the two periods showed that there was no difference in the proportion of patients with four pregnancy-specific liver diseases （hyperemesis gravidarum， preeclampsia and eclampsia， intrahepatic cholestasis of pregnancy， and acute fatty liver of pregnancy）. As for the patients with abnormal liver biochemical parameters in pregnancy， there was no significant difference in the proportion of patients with normal creatinine and stimated glomerular filtration rate （eGFR） between the epidemic period and the non-epidemic period （86.78% vs 87.90%， χ²=0.141， P=0.708）. The patients with ALT≥5×upper limit of normal accounted for 7.94% in the epidemic period of COVID-19 and 9.54% in the non-epidemic period （χ²=0.433， P=0.511）， and the patients with severe cholestasis accounted for 7.75% in the epidemic period of COVID-19 and 9.27% in the non-epidemic period （χ²=0.392， P=0.531）. The proportion of patients with obstetric bleeding during the epidemic period of COVID-19 was significantly lower than that during the non-epidemic period （14.61% vs 24.19%， χ²=489.334， P<0.001）. ConclusionThere is no difference in the proportion of patients with pregnancy-specific liver diseases among the patients with abnormal liver biochemical parameters in pregnancy between the epidemic period and the non-epidemic period of COVID-19， and there is no change in the proportion of patients with normal creatinine and eGFR among these patients in the epidemic period of COVID-19.

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.

Objective:To improve the ability of radiation health technical institutions for biological dose estimation.Methods:A total of 144 institutions nationwide, including the CDC, prevention and treatment center for occupational disease, colleges and universities, scientific research institutes, nuclear industry systems, and medical and physical examination institutions, were organized to carry out the intercomparison of national biological dose estimation capabilities in 2022. The institutions participating in the comparison were divided into two types of A and B, through the identification of chromosome aberrations, to estimate the irradiation dose (A) or chromosome aberration rate (B). The results were summarized and compared, and the main problems were analyzed and discussed.Results:There were 60 institutions in type A, 52 qualified institutions (including 12 excellent institutions) and 8 unqualified institutions, with a pass rate of 86.7% (20.0% excellent) and a failure rate of 13.3%. There were 84 institutions participating in the biological dose estimation comparison of type B, with 48 qualified institutions, and 36 unqualified institutions, the qualified rate was 57.1%, and the unqualified rate was 42.9%.Conclusions:Most of the institutions participating in type A comparison have the ability to estimate biological dose, and more than half of the institutions participating in type B comparison have the ability to analyze chromosome aberration. The overall ability of institutions participating in type A comparison is higher than in type B.

Objective To investigate the effects of prolonged low-dose neutron-γ radiation on peripheral blood lymphocytes of logging workers. Methods The health information of workers in a logging company was collected by on-site blood sample collection and questionnaire survey. Individual doses of γ and neutron radiation were recorded using LiF elements and CR-39, respectively. Lymphocyte count in peripheral blood was measured by blood cytometer. Cell cycle and cyclins were detected by flow cytometry. Results The annual dose of some logging workers exceeded 5 mSv. Lymphocyte counts showed a difference of 15% between the group exposed to the lowest annual dose of 0–1 mSv (mean: 2.45 × 10⁹/L) and the group exposed to the highest annual dose of 5–25 mSv (mean: 2.08 × 10⁹/L). In comparison to pre-shift workers, logging workers exhibited a G1-phase arrest in the lymphocyte cycle, along with increased expression of cyclins p21 and CDK2. Conclusion Prolonged exposure to low-dose neutron-γ radiation leads to reduced lymphocyte counts as well as changes in lymphocyte cycle and cyclin expression.

Objective:To construct a novel dual-specific antibody targeting human CD123 (CD123 DuAb) and study its effects in acute myeloid leukemia (AML) .Methods:Based on the variable region of the CD123 monoclonal antibody independently developed at our institution, the CD123 DuAb expression plasmid was constructed by molecular cloning and transfected into ExpiCHO-S cells to prepare the antibody protein. Through a series of in vitro experiments, its activation and proliferation effect on T cells, as well as the effect of promoting T-cell killing of AML cells, were verified.Results:① A novel CD123 DuAb plasmid targeting CD123 was successfully constructed and expressed in the Expi-CHO eukaryotic system. ②The CD123 DuAb could bind both CD3 on T cells and CD123 on CD123 + tumor cells. ③When T cells were co-cultured with MV4-11 cells with addition of the CD123 DuAb at a concentration of 1 nmol/L, the positive expression rates of CD69 and CD25 on T cells were 68.0% and 44.3%, respectively, which were significantly higher than those of the control group ( P<0.05). ④Co-culture with CD123 DuAb at 1 nmol/L promoted T-cell proliferation, and the absolute T-cell count increased from 5×10 5/ml to 3.2×10 6/ml on day 9, and CFSE fluorescence intensity decreased significantly. ⑤ With the increase in CD123 DuAb concentration in the culture system, T-cell exhaustion and apoptosis increased. When the CD123 DuAb was added at a concentration of 1 nmol/L to the culture system, the proportion of CD8 + PD-1 + LAG-3 + T cells was 10.90%, and the proportion of propidium iodide (PI) - Annexin Ⅴ + T cells and PI + Annexin Ⅴ + T cells was 18.27% and 11.43%, respectively, which were significantly higher than those in the control group ( P<0.05). ⑥ The CD123 DuAb significantly activated T cells, and the activation intensity was positively correlated with its concentration. The expression rate of CD107a on T cells reached 16.05% with 1 nmol/L CD123 DuAb, which was significantly higher than that of the control group ( P<0.05). ⑦The CD123 DuAb promoted cytokine secretion by T cells at a concentration of 1 nmol/L, and the concentration of IFN-γ and TNF-α in the supernatant of the co-culture system reached 193.8 pg/ml and 169.8 pg/ml, respectively, which was significantly higher than that of the control group ( P<0.05). ⑧When CD123 DuAb was added at a concentration of 1 nmol/L to the co-culture system of T cells and CD123 + tumor cells, the killing intensity of T cells significantly increased, and the residual rates of CD123 + MV4-11 cells, CD123 + Molm13 cells, and CD123 + THP-1 cells were 7.4%, 6.7%, and 14.6% on day 3, respectively, which were significantly lower than those in the control group ( P<0.05) . Conclusion:In this study, a novel CD123 DuAb was constructed and expressed. In vitro experiments verified that the DuAb binds to CD123 + tumor cells and T cells simultaneously, promotes T-cell activation and proliferation, and facilitates their anti-leukemia effect, which provides a basis for further clinical research.

Objective:To summarize the clinical features of epilepsy and (or) developmental delay associated with KCNB1 gene variants in children.Methods:A case series study was conducted on 24 children with KCNB1 gene variants associated with epilepsy and (or) developmental delay who were treated at the Children′s Medical Center of Peking University First Hospital and the Department of Neurology of Shenzhen Children′s Hospital from July 2015 to June 2024. The manifestations of seizures, electroencephalogram (EEG) and genetic test results of those children were analyzed.Results:All the KCNB1 gene variants were de novo, involving 20 different variation, including 15 missense variations, 3 frameshift variations and 2 nonsense variations. There were 7 novel variations. Among the 24 developmental and epileptic encephalopathy children, there were 14 boys and 10 girls. The last follow-up age ranged from 9 months to 13 years and 9 months. Seizures were present in 21 children (88%), with onset ranging from 1 month to 7 years, and 76% (16/21) began before 2 years of age. The seizure types included focal seizures in 15 children (71%), epileptic spasms, myoclonic seizures, and generalized tonic-clonic seizures in 6 children respectively, atypical absence seizures in 4 children, and myoclonic atonic seizures in 1 child. Seventeen children (81%) had a cluster of seizures and 5 had a history of focal status epilepticus with impaired consciousness. All 24 children had varying degrees of developmental delay, with 3 presenting solely developmental delay. EEG abnormalities were present in all the 21 children with seizures, including focal or multifocal discharges in 20 children, generalized discharges in 10 children, hypsarrhythmia in 2 children, and electrical status epilepticus during sleep in 3 children. Magnetic resonance imaging abnormalities were found in 5 of the 24 children. Among the 21 children with seizures, 57% (12/21) achieved seizure control.Conclusions:KCNB1 gene variants are predominantly de novo missense variation. Most affected children present with epilepsy, though some may exhibit only developmental delay. Epilepsy often begins before 2 years of age, with focal seizures being the most common type. About 80% of patients experience clustered seizures. Although most patients achieve seizure control, they still exhibit varying degrees of developmental delay, consistent with developmental epileptic encephalopathy.

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.