BACKGROUND:Psoralen has a strong anti-osteoporotic activity and may have a restorative effect on chemotherapy-induced osteoporosis. OBJECTIVE:To explore the restorative effect of psoralen on the osteogenic differentiation of bone marrow mesenchymal stem cells in mice inhibited by cyclophosphamide and its mechanism. METHODS:C57BL/6 mouse bone marrow mesenchymal stem cells were isolated and cultured.Effect of psoralen on viability of bone marrow mesenchymal stem cells was detected by MTT assay.Osteogenic induction combined with alkaline phosphatase staining was used to determine the optimal dose of psoralen to restore the osteogenic differentiation of bone marrow mesenchymal stem cells inhibited by cyclophosphamide.The mRNA expression levels of Runx2,alkaline phosphatase,Osteocalcin,osteoprotegerin,and Wnt/β-catenin signaling pathway-related genes Wnt1,Wnt4,Wnt10b,β-catenin,and c-MYC were measured by RT-qPCR at different time points under the intervention with psoralen.The protein expression of osteogenic specific transcription factor Runx2 and Wnt/β-catenin signaling pathway related genes Active β-catenin,DKK1,c-MYC,and Cyclin D1 was determined by western blot assay at different time points under the intervention with psoralen. RESULTS AND CONCLUSION:(1)There was no significant effect of different concentrations of psoralen on the viability of bone marrow mesenchymal stem cells.The best recovery of the inhibition of osteogenic differentiation of bone marrow mesenchymal stem cells caused by cyclophosphamide was under the intervention of psoralen at a concentration of 200 μmol/L.(2)Psoralen reversed the reduction in osteogenic differentiation marker genes Runx2,alkaline phosphatase,Osteocalcin and osteoprotegerin mRNA expression and Runx2 protein expression in bone marrow mesenchymal stem cells caused by cyclophosphamide conditioned medium.(3)Psoralen reversed the decrease in Wnt/β-catenin pathway-related genes Wnt4,β-catenin,c-MYC mRNA and Active β-catenin,c-MYC,and Cyclin D1 protein expression and the increase in DKK1 protein expression in bone marrow mesenchymal stem cells caused by cyclophosphamide conditioned medium.(4)The results showed that cyclophosphamide inhibited osteogenic differentiation of bone marrow mesenchymal stem cells in mice,and psoralen had a restorative effect on it.The best intervention effect was achieved at a concentration of 200 μmol/L psoralen,and this protective effect might be related to the activation of Wnt4/β-catenin signaling pathway by psoralen.

Iron overload is strongly associated with heart disease. Ferroptosis is a new form of regulated cell death indicated in cardiac ischemia-reperfusion (I/R) injury. However, the specific molecular mechanism of myocardial injury caused by iron overload in the heart is still unclear, and the involvement of ferroptosis in iron overload-induced myocardial injury is not fully understood. In this study, we observed that ferroptosis participated in developing of iron overload and I/R-induced cardiomyopathy. Mechanistically, we discovered that Parkin inhibited iron overload-induced ferroptosis in cardiomyocytes by promoting the ubiquitination of long-chain acyl-CoA synthetase 4 (ACSL4), a crucial protein involved in ferroptosis-related lipid metabolism pathways. Additionally, we identified p53 as a transcription factor that transcriptionally suppressed Parkin expression in iron-overloaded cardiomyocytes, thereby regulating iron overload-induced ferroptosis. In animal studies, cardiac-specific Parkin knockout mice (Myh6-CreER ^T2 /Parkin ^fl/fl ) fed a high-iron diet presented more severe myocardial damage, and the high iron levels exacerbated myocardial I/R injury. However, the ferroptosis inhibitor Fer-1 significantly suppressed iron overload-induced ferroptosis and myocardial I/R injury. Moreover, Parkin effectively protected against impaired mitochondrial function and prevented iron overload-induced mitochondrial lipid peroxidation. These findings unveil a novel regulatory pathway involving p53-Parkin-ACSL4 in heart disease by inhibiting of ferroptosis.

Interferon regulatory factor 4 (IRF4) is a critical transcription factor that governs the differentiation of cluster of differentiation 4⁺ (CD4⁺) T cells. The pathogenesis and progression of psoriasis are primarily attributed to an immune imbalance stemming from the overproduction of interleukin-17A (IL-17A) by T lymphocytes. However, the role of IRF4 in psoriasis remains unexplored. In this study, we found that IRF4 activity is increased in the cutaneous lesions of patients with psoriasis in response to stimulation by IL-23A and IL-1β. This IRF4 elevation heightens its binding to the E1A binding protein p300 (EP300) promoter, triggering the transcription of downstream retinoic acid receptor-related orphan receptor-γt (RORγt) and increasing the secretion of IL-17A, thereby establishing the IL-1β/IL-23A-IRF4-EP300-RORC-IL-17A inflammatory cascade in psoriasis. The alleviation of imiquimod (IMQ)-induced psoriatic-like symptoms was achieved through the creation of a Irf4 ^-/- gene deletion mouse model and pharmacological inhibition using antisense oligonucleotides targeted for Irf4. This amelioration was accompanied by a decreased number of IL-17A-producing CD4⁺ T cells in the skin. The findings of this study suggest that IRF4 plays a crucial role in the promotion of inflammation and exacerbation of IMQ-induced psoriasiform dermatitis. Consequently, IRF4 targeting could be a promising therapeutic strategy.

Objective: To understand the association between beverage and snack intake and insufficient serum 25 hydroxyvitamin D [25(OH)D] among primary and secondary school students, so as to provide a scientific basis for targeted intervention measures. Methods: From October to December 2021, a stratified random sampling method was used to select 2 477 primary and secondary school students aged 8 to 15 years old from 9 counties in Yunnan Province implemented the Nutrition Improvement Plan for Rural Compulsory Education Students. The intake of beverages and snacks was investigated using the Rural Student Nutrition Monitoring Questionnaire from Chinese Center for Disease Control and Prevention. The snack intake intensity was calculated and classified into no intake, extremely low, low, medium, and high intensity. Serum 25(OH)D levels were measured in the laboratory, and levels <20 ng/mL were defined as insufficient. Chi square tests, LASSO regression, random forest and binary Logistic regression were used to analyze the association between 20 types of beverages and snacks and serum 25(OH)D insufficiency. Results: Insufficient serum 25(OH)D was detected in 564 boys (45.9%) and 855 girls (68.5%), with a total of 1 419 cases (57.3%). Binary Logistic regression results showed that extremely low intake intensity of carbonated beverages ( OR =1.51), plant protein beverages ( OR =1.61), and milk tea beverages ( OR =1.39) increased the risk of insufficient serum 25(OH)D, while protective factors were fruits and vegetables ( OR =0.77) and pure milk and yogurt ( OR =0.74) (all P <0.05). Subgroup analysis showed that extremely low intake intensity of carbonated beverages, milk containing beverages, tea beverages, fruit and vegetable juices, and plant protein beverages increased the risk of insufficient serum 25(OH)D in girls ( OR =2.22, 1.72, 1.67, 1.74, 1.92), and high intake intensity increased the risk of insufficient serum 25(OH)D in boys ( OR =1.73, 1.48, 1.52, 1.49, 1.97) (all P <0.05). Extremely low intake intensity of carbonated beverages, plant protein beverages, and milk tea beverages in junior high school students ( OR =1.92, 2.54, 1.68) and low intake intensity in primary school students ( OR =1.40, 1.33, 1.45) increased the risk of insufficient serum 25(OH)D (all P <0.05). Conclusions Frequent intake of beverages and highly processed snacks increases the risk of insufficient serum 25(OH)D in primary and secondary school students, while natural foods such as fruits, vegetables, pure milk and yogurt can reduce the risk. Girls and junior high school students are more susceptible to these effects.

ObjectivesTo determine the syphilis infection and risk factors among the male outpatients in sexually transmitted disease （STD） clinics in Taizhou， and to provide a scientific evidence for the development of comprehensive prevention and intervention strategies for syphilis. MethodsA questionnaire survey and serological testing were conducted among the male STD outpatients in the hospitals designated for STD treatment in Jiaojiang， Linhai， Luqiao and Sanmen in Taizhou. Logistic regression analysis was used to determine the risk factors associated with syphilis infection. ResultsA total of 16 400 male STD outpatients were investigated， with a syphilis infection rate of 2.10%. Multivariate analysis showed that aged over 60， non-local household registration， engaging in sexual activity with commercial sex workers in the past three months， having two or more sexual partners in the past three months， engaging in anal sex with heterosexual partners， co-infection with human immunodeficiency virus （HIV）， and co-infection with hepatitis C virus （HCV） were significantly associated with a higher syphilis infection rate （all P<0.05）. Knowledge of HIV/AIDS， a history of chlamydia infection in the past year， and a history of genital wart infection were significantly associated with a lower syphilis infection rate （all P<0.05）. ConclusionSyphilis infection among the male STD outpatients in Taizhou remained lower， compared with the national average level. Targeted comprehensive prevention and control strategies should be implemented based on relevant risk factors. Screening and prevention interventions should be prioritized for the individuals with non-local household registration， at-risk sexual behavior， and co-infection with HCV and HIV. Strengthening the collaborative efforts is also recommended for joint prevention and control of HIV/AIDS and syphilis.

Objective To compare the value of TACE+microwave ablation(MWA)+programmed death-1(PD-1)and TACE+MWA for treating primary hepatic carcinoma(PHC).Methods Data of 80 PHC patients who underwent TACE+MWA+PD-1(observe group)or TACE+MWA treatment(control group)were retrospectively analyzed(each n=40).The baseline data,therapeutic efficacy of PHC and complications were compared between groups.Results No significant difference of baseline data was found between groups(all P＞0.05).The objective response rate(ORR)of observe group and control group was 90.00％and 72.50％,respectively,while the disease control rate(DCR)was 97.50％and 95.00％,respectively,with no significant difference between groups(both P＞0.05).The median overall survival(OS)of observe group and control group was 30.80 months and 15.70 months,respectively,while the median progression free survival(PFS)was 23.35 months and 6.80 months,respectively.OS and PFS of observe group were both longer than those of control group(both P＜0.05).No significant difference of the incidence of complications was detected between groups(all P＞0.05),and all were improved after symptomatic treatment.Conclusion TACE+MWA+PD-1 was superior to TACE+MWA for treating PHC.

Objective:To examine the satisfaction and teaching effectiveness of Health Statistics among students majoring in Public Utility Administration,and to provide valuable insights for optimizing current teaching practices and enhancing teaching effectiveness.Methods:A survey questionnaire,specifically designed for this study,was administered to 56 students of grade 2021 majoring in Public Utility Administration in our school.The questionnaire covered topics such as the difficulty of teaching content,learning satisfaction,and the Student Evaluation of Educational Quality(SEEQ)scale.Results:Non-parametric testing was considered the most difficult knowledge point,and male students found statistical inference of quantitative data was the most challenging,while female students found non-parametric testing was the most difficult.In terms of satisfaction,the highest score was obtained for pre-class review,while the lowest score was given to improving learning interest in Rain Classroom.The overall score for teaching effectiveness,as measured by SEEQ,was good across seven dimensions.Teaching enthusiasm received the highest score(4.40±0.43),while homework volume received the lowest score(3.23±0.37).With the exception of homework grading,satisfaction was positively correlated with most dimensions of teaching effectiveness evaluation.Conclusions:Despite perceiving the course as generally difficult,students expressed high satisfaction and overall positive evaluation of teaching effectiveness.Therefore,it is recommended to actively explore new teaching reforms to cultivate well-rounded applied talents for public utilities.

OBJECTIVE To establish and validate a rat liver micronucleus test(LMNT)method based on fixation of liver tissue with 4%neutral formaldehyde(HCHO fixation)for preparation of hepa-tocytes(HEPs).METHODS ①The LMNT based on neutral HCHO fixation(HCHO fixation-LMNT)was established using the liver micronucleus positive compound N-nitrosodiethylamine(DEN).SD rats were divided into female and male groups,and each group was randomly subdivided into the vehicle control group and DEN 12.5 mg·kg-1 group,with five rats in each.The rats were ig administered with normal saline and DEN once a day for 14 consecutive days,after which liver tissues were collected.Some of the tissue was digested with collagenase to prepare HEP suspension,and the remaining tissue was used to prepare HEP suspension with HCHO fixation.After staining with SYBR Gold,the number of micronucleated hepatocytes(MN-HEP)and the number of HEPs in the mitotic phase were counted under a microscope.The micronucleus rate of HEP(MN-HEP rate)and the mitotic index were calculated,and an MN-HEP rate>0.07%was considered positive.②Male SD rats were divided into the quinoline(30,60,120 mg·kg-1)group,N-nitrosoopyrrolidine(NPYR,25,50,100 mg·kg-1)group,vehicle control group(deionized water for NPYR,and corn oil for quinoline),and positive control DEN(12.5 mg·kg-1)group,with 5-6 rats per group,and were ig administrated for 15 consecutive days.Body mass was recorded daily,and at the end of the experiment,the liver was removed to record the total liver weight,and calculate the liver coefficient.Liver function-related serum biochemical indicators including glutamic-pyruvic transaminase(GPT),glutamic-oxaloacetic transaminase(GOT)activities,and levels of total bili-rubin(T-BIL)were measured and direct bilirubin(D-BIL)using an automatic biochemical analyzer.The MN-HEP rate was determined using the collagenase digestion and HCHO fixation methods,and the peripheral blood MN assay and hepatocellular carcinoma comet assay were conducted to evaluate the genotoxicity of quinoline and NPYR.RESULTS ① Compared with the corresponding vehicle control groups(0.069%and 0.030%),the MN-HEP rate of male rats treated with DEN by formalin-LMNT was 1.10%,and the MN-HEP rate of female ones was 0.82%,both significantly increased(P<0.05).Com-pared with corresponding vehicle control groups(0.060%and 0.030%),the MN-HEP rate of male rats treated with DEN by collagenase digestion-LMNT was 1.45%,and that of female rats was 0.46%,both significantly increased(P<0.05),which were considered positive.The MN-HEP rate of male rats was significantly higher than that of females with both methods(P<0.05).There was no significant differ-ence in mitotic indexes between the DEN groups by collagenase digestion-LMNT and HCHO fixation-LMNT in male and female rats compared to corresponding vehicle control groups.② Compared to the vehicle control group,the body mass of rats in the NPYR 50 and 100 mg·kg-1 groups was significantly reduced 7 to 14 days into the ig administration(P<0.01),and the DEN group showed a significant reduction at days 8 to 14(P<0.01).The body mass of rats in the quinoline 120 mg·kg-1 group was signifi-cantly reduced 4 to 14 days into the ig administration(P<0.01),and the DEN group showed a signifi-cant reduction at days 10 to 14(P<0.05).Compared to the vehicle control group,both the liver weight and liver coefficient were significantly reduced in the NPYR 100 mg·kg-1 group(P<0.01)and the DEN group(P<0.05).The liver weight(P<0.01)and liver coefficient(P<0.05)were significantly increased in the quinoline 60 and 120 mg·kg-1 groups.Compared to the vehicle control group,the serum T-BIL level was significantly increased in the DEN group(P<0.01),and the activities of GPT and GOT,as well as the levels of D-BIL and T-BIL,were significantly increased in the NPYR 100 mg·kg-1 group(P<0.01).There were no significant changes in the NPYR 25,50 mg·kg-1 groups or any of the dose groups of quinoline.The MN-HEP rate by HCHO fixation-LMNT for NPYR was slightly higher than that by collage-nase digestion-LMNT,both considered positive.Compared with corresponding control group,the MN-HEP rate by formalin-LMNT for NPYR and the MN-HEP rate by collagenase digestion for NPYR were both significantly increased(P<0.05).The MN-HEP rate by HCHO fixation-LMNT for quinoline was comparable to that by collagenase digestion-LMNT,both considered positive.Compared with corresponding vehicle control group,the MN-HEP rate by HCHO fixation-LMNT for quinoline and the MN-HEP rate by collagenase digestion-LMNT for quinoline were both significantly increased(P<0.05).The correlation between the MN-HEP rates based on HCHO fixation and collagenase digestion for NPYR and quinoline was good(R2=0.8614 and 0.9279,respectively).In the NPYR groups,the periph-eral blood micronucleus assay were negative,while the comet assay results were positive.In the quino-line group,both the peripheral blood micronucleus assay and the comet assay results were negative.CON-CLUSION The HCHO fixation-LMNT has been established and validated,and the sensitivity of the LMNT method based on HCHO fixation-LMNT for detection of hepatocarcinogens is higher than that of collagenase digestion-LMNT.

Chinese materia medica and its compound formulas have the characteristics of multi-component,multi-target,and multi pathway effects,and have unique advantages in preventing and treating complex diseases.Network pharmacology explains disease patterns and drug mechanisms from the perspective of complex biological networks,and explores the pharmacological substance basis and target of Chinese medicine.This article summarized the application of network pharmacology from the aspects of biological basis of TCM syndrome,the substance basis and pharmacological mechanism of TCM,compatibility theory of TCM compound formulas,etc.,with the purpose to provide a reference for the research and application of network pharmacology in TCM.

Objective:To explore the produced-radiation brain damage in simulated solar particle events and to provide evidence for health risk assessment of radiation from manned deep space exploration.Methods:According to the main characteristics of solar particle events, mice were treated with total body irradiation (TBI) with 90 MeV protons in a dose range from 0.1 to 2 Gy, with irradiation dose of 0, 0.1, 0.3, 0.5, 1, 2 Gy, respectively. At 3 and 7 d after irradiation, the behavior of mice was examined using balance beam tests, rotarod tests, and new object recognition tests. Then, the density of dendritic spines and the number of Nissl bodies in the hippocampus were measured using Golgi and Nissl staining. The superoxide dismutase (SOD) activity, malondialdehyde (MDA) content, and neurotransmitter content in brain tissue were detected using the WST-8 method, TBA method, and high pressure liquid chromatography (HPLC), respectively. Besides, cell apoptosis was determined using the TUNEL method, and the dose-response relationship, a function of dose change with damage index, was analyzed using linear and linear square fitting method. Finally, the minimum radiation dose causing a significant change in all indicators of brain damage was determined as the brain damage threshold.Results:Compared to the control group, 1 Gy proton irradiation result ed in a significant decrease in the density of filopod dendritic spines ( t = 1.82, 2.30, P < 0.05) and a significant increase in abnormal Nissl bodies in the CA1 region ( t = 2.44, 3.77, P < 0.05). At 3 and 7 d after irradiation, as well as a significant increase in the DA ( t = 2.52, P<0.05) and Glu contents ( t = 4.04, P < 0.05) on day 7. In contrast, 2 Gy proton irradiation result ed in a decrease in SOD activity on day 3 ( t = 3.44, P < 0.05), and an increase in the MDA content ( t = 1.90, 2.14, P < 0.05), hippocampal cell apoptosis (t = 3.91, 3.54, P < 0.05), and 5-HT levels ( t = 2.81, 2.69, P < 0.05), together with a decrease in climbing time in the rotarod tests ( t = 2.85, 2.64, P<0.05) and propensity to recognize new objects ( t = 2.87, 2.84, P < 0.05) on days 3 and 7. Furthermore, a dose-response relationship was observed in the dose range from 0.1 to 2 Gy ( R2=0.74-0.99). Conclusions:The dose threshold of 90 MeV protons inducing brain damage in mice is inferred to be 1 Gy, and 14 dose-response models are developed, providing a biological basis for organ dose capping and risk assessment of crew experiencing short-term deep space flights.