ObjectiveThe malaria parasites remodel the host erythrocyte structure by exporting parasite proteins that interact with the membrane skeleton proteins of red blood cells (RBCs), facilitating their intracellular survival and pathogenicity. Skeleton-binding protein 1 (SBP1) is a conserved exported protein across Plasmodium species. In Plasmodium falciparum, SBP1 has been reported to interact with erythrocyte membrane skeleton proteins 4.1R and spectrin, while its contribution to erythrocyte remodeling and parasite virulence in Plasmodium berghei (Pb) remains unclear. This study aims to determine whether PbSBP1 associates with the host cytoskeletal protein 4.1R and to investigate its role in the remodeling of host RBCs and the pathogenicity of Plasmodium berghei. MethodsIn Plasmodium berghei, the relationship between PbSBP1 and the erythrocyte cytoskeletal protein 4.1R was examined using co-immunoprecipitation. A Pbsbp1 gene knockout mutant of Plasmodium berghei (Pbsbp1∆) was generated based on the principle of double crossover homologous recombination. The deformability of erythrocytes infected with Pbsbp1∆ parasites was assessed using microfluidic methods. Microchannels with an array of cylindrical pillars were used to detect modifications in infected RBC deformability. The infected RBCs were squashed between the rows and recovered between the columns and the transit velocity (μm/s) of infected RBCs travelling through the microchannel was recorded. The component of the erythrocyte membrane skeleton junctional complex, tropomodulin (TMOD), was fluorescently labeled, and the cytoskeletal network of infected erythrocytes was imaged using super-resolution stochastic optical reconstruction microscopy (STORM) to analyze ultrastructural changes in the cytoskeleton of wild-type (WT) and Pbsbp1∆-infected erythrocytes. Actin-based junctional complexes were displayed as individual clusters by the labeled TMOD in the STORM images, and the cluster densities and distances between adjacent clusters of infected RBCs were calculated. Additionally, rodent malaria models (BALB/c mice) and experimental cerebral malaria models (C57BL/6 mice) were employed to monitor the growth of Pbsbp1∆ and WT parasites during the intraerythrocytic stage and their capacity to induce cerebral malaria in mice. ResultsPbSBP1 may participate in the remodeling of infected erythrocytes through direct or indirect interaction with the erythrocyte cytoskeletal protein 4.1R. Microfluidic assays revealed that the deformability of erythrocytes infected with Pbsbp1∆ parasites was significantly enhanced compared to those infected with WT parasites. STORM imaging further demonstrated that the ultrastructure of the erythrocyte cytoskeleton in Pbsbp1∆-infected cells was altered relative to that in WT-infected erythrocytes. The distances between nearest neighbors of clusters had a tendency to increase while the cluster densities were decreased in Pbsbp1∆-infected RBCs compared to WT-infected RBCs. Subsequent phenotypic analysis indicated that the growth rate of Pbsbp1∆ parasites during the intraerythrocytic stage was significantly slower than that of WT parasites, and their ability to induce cerebral malaria in mice was also attenuated. These findings suggest that PbSBP1 is involved in the remodeling of the erythrocyte membrane skeleton, likely through its direct or indirect interaction with protein 4.1R, thereby regulating the deformability of infected erythrocytes and influencing the pathogenicity of the blood-stage parasites. ConclusionThis study establishes a role for PbSBP1 in host erythrocyte remodeling and parasite virulence, providing new research strategies for the prevention and treatment of malaria.

Purpose: In this study, the neural communication patterns between hypothalamic structures and cortical areas in girls diagnosed with central precocious puberty (CPP) were explored. Endocrine profiles were incorporated to clarify the pathophysiological interactions between cerebral networks and hormonal regulation. The hypothalamus was designated as the key focus area for connectivity analysis. Methods: Fifty-seven girls (37 CPP, 20 non-CPP) were recruited from the Pediatric Development Clinic at the Second Affiliated Hospital of Anhui Medical University. The collected data included demographic information, gonadotropin-releasing hormone stimulation tests, and magnetic resonance imaging scans. Hypothalamic functional connectivity (FC) was analyzed using predefined region of interest coordinates, and correlations between hormone levels and FC values were assessed. Results: Compared to the non-CPP group, the CPP group exhibited elevated baseline follicle-stimulating hormone (FSH), baseline luteinizing hormone (LH), peak FSH, peak LH, and peak LH/FSH ratios. Patients with CPP exhibited enhanced neural synchronization linking the right lateral hypothalamic effector zone to the right superior frontal gyrus (displaying a borderline significant correlation with peak LH concentrations), concurrent with diminished functional coupling of the right lateral hypothalamic efferent to the right fusiform/supramarginal gyri. The left lateral hypothalamic projections demonstrated amplified connectivity with the right cuneus. No FC differences were observed in the medial hypothalamus. Conclusion Abnormal lateral hypothalamus FC patterns were identified in CPP girls and were particularly linked to peak LH levels. The findings offer novel insights into the neuroendocrine mechanisms underlying CPP.

Laboratory animal science is an emerging interdisciplinary field supporting life science and medical research, and a key component of talent cultivation and technological innovation in medical colleges. Currently, this field faces challenges such as a significant imbalance between talent supply and demand and a weak systematic training system. To respond to national strategies and societal needs, Shandong First Medical University established the School of Laboratory Animal through industry-education integration during the 14th Five-Year Plan period. The school is based on The Medical Laboratory Technology major, cultivates application-oriented, interdisciplinary professionals in laboratory animal science, and has constructed a three-in-one training system integrating "course learning–scientific research–industry practice". Specific measures for talent cultivation include: optimizing the general education, professional courses, and intensive practical modules in the talent training program, and establishing the "Yellow River Class" integrating industry and education; implementing an undergraduate mentor system, and conducting scientific research training based on The Model Animal Research and Development Engineering Laboratory; collaborating with bases across the entire industrial chain of laboratory animal production, research, application, and quality control, appointing industry mentors, and strengthening practical teaching. Practice has shown that this system has been remarkably effective: a total of 320 undergraduate students have been enrolled since 2020; the employment rate of undergraduate graduates for two consecutive cohorts has reached 100%, and the postgraduate enrollment rate has exceeded 50%; undergraduate students have won numerous national and provincial awards in academic competitions, and have obtained multiple patents and published papers. In the future, the school will further integrate the advantages of medicine, agriculture, and science, optimize the depth and breadth of courses, strengthen the construction of faculty and teaching materials, and improve the undergraduate-master's integrated training mechanism. This article can provide a reference for the training of professionals in laboratory animal science and related biomedical fields in medical colleges.

Objective This study reports on the obligatory parasitism of water mites Arrenurus sp. on Anopheles messeae at the Manzhouli Port, Inner Mongolia, China. Methods Duing July 2024, a survey on the mosquito diversity was conducted at the Manzhouli Port. Captured mosquitoes and their ectoparasites were identified to species level. Results A total of 1840 adult mosquitoes were collected, representing species from three genera: Culex(Cx. modestus, Cx. pipiens pallens), Aedes(Ae. dorsalis, Ae. flavidorsalis, Ae. flavescens), and Anopheles (An. messeae). Among all the mosqutioes specimens,3 out of 150 captured An. messeae were found to carry ectoparasitic mites, with number of 2,4,27 mites separately. Morphological and molecular identification reached the same result as water mites(Hydrachnidiae, Hydracrina). COI gene sequence showed 94% similarity with the closest species Arrenurus truncatellus. Conlusions Literature review suggests water mites are host-specific parasitism of mosquito species and herein with the first record of Arrenurus sp. parasiting on An. Messeae in the most high-latitude region globally.

Objective To develop an erythrocyte-based delivery system for butyrylcholinesterase(BChE)that is capable of prophylaxis against organophosphorus nerve agents.Methods Recombinant BChE was produced and analyzed for oligomerization via polyacrylamide gel electrophoresis(PAGE)and Western blotting.A modified hypotonic preswelling method was employed to prepare BChE-loaded erythrocytes.The drug loading capacity and encapsulation efficiency were quantified using enzyme-linked immunosorbent assay(ELISA).Catalytic activity was assessed in vitro with an activity detection kit.The system was characterized via scanning electron microscopy(SEM),flow cytometry and a hematology analyzer.Results Recombinant BChE predominantly existed as dimers(85％dimer,15％monomer).The optimized volume ratio of erythrocytes to hypotonic solution was determined as 1:7.Compared with native and empty erythrocytes,BChE-loaded erythrocytes exhibited significantly higher catalytic activity(P＜0.001).The mean corpuscular volume of BChE-loaded erythrocytes increased(P＜0.001),while the mean content of corpuscular hemoglobin and hemoglobin in erythrocytes per 100 mL decreased(P＜0.001).SEM revealed no morphological differences(biconcave disc shape).Hypotonic preswelling moderately increased erythrocyte apoptosis(P＜0.001),but no statistical difference was observed between BChE-loaded and hypotonic-treated erythrocytes(P＞0.05).CD47 expression remained unchanged compared to native erythrocytes(P＞0.05).Conclusion The modified hypotonic preswelling method can generate BChE-loaded erythrocytes that retain the characteristics of native erythrocytes while conferring catalytic activity,offering a novel strategy for clinical intervention against organophosphorus poisoning.

A mounting body of research suggests that circRNAs significantly contribute to the develop-ment of myocardial fibrosis.The microarray results of human circular RNA expression profile indicated that circHERC4_041 expression increased in the myocardium of patients with heart failure,RT-qPCR a-nalysis confirmed that the myocardial expression level of circHERC4_041 in individuals with heart failure were considerably elevated compared to that in healthy organ donors.Fluorescence in situ hybridization(FISH)confirmed that circHERC4_041 was abundant in the cytoplasm of human cardiomyocyte AC16.Overexpression of circHERC4_041 in mouse myocardial fibroblasts(mCFs)mediated by adenovirus in-hibited the expression of fibrosis-related proteins in mCFs.Experiments involving cell proliferation,wound healing,and Transwell assays demonstrated that overexpression of circHERC4_041 suppressed the growth and mobility of mCFs(P＜0.001).Sequence analysis results suggested that circHERC4_041 con-tains potential ribosome entry sequence(IRES)and open reading frame(ORF).Western blot confirmed that circHERC4_041 could translate the 516 amino acid HERC4-516aa protein,which was mainly located in the cytoplasm of the cell.Cell functional experiments confirmed that circHERC4_041 inhibited the fi-brotic phenotype of mCFs by specifically translating HERC4-516aa(P＜0.05).The specific interaction between HERC4-516aa and transglutaminase 2(TGM2)was confirmed by IP-MS screening and Co-IP i-dentification.Further results found that the degradation of TGM2 was promoted through proteasome path-way.The overexpression of TGM2 in mCFs facilitated by adenoviral vectors could counteract the suppres-sive effects of HERC4-516aa on the fibrotic phenotype of mCFs.Therefore,this study confirmed that the HERC4-516aa protein translated by circHERC4_041 can specifically bind to TGM2 to inhibit the fibrotic phenotype of myocardial fibroblasts.

Neuroblastoma(NB),the most common type of extracranial solid tumor in children,is char-acterized by high malignancy and poor prognosis,warranting in-depth investigation.In recent years,mi-croRNAs(miRNAs)have emerged as crucial post-transcriptional regulators playing pivotal roles in tu-morigenesis and progression.Building upon this background,the present study specifically focuses on in-vestigating miR-3910's biological functions and underlying molecular regulatory mechanisms in the NB SH-SY5Y cell line.Through bioinformatics analysis and transcriptome sequencing,we identified potential key target molecules of miR-3910,thereby providing genetic targets for the precise diagnosis and effective treatment of NB.In this study,qRT-PCR was employed to measure miR-3910 expression levels in SH-SY5Y cells transfected with mimic negative control and miR-3910 mimic.Compared to the nc group,miR-3910 expression was significantly upregulated in the mimic group(P＜0.01).The CCK-8 assay and scratch wound healing assay were used to quantitatively assess the impact of miR-3910 on cell prolif-eration and migration.Results showed that cell proliferation significantly increased at 48 h(P＜0.05),and migration ability was markedly enhanced at 48 h(P＜0.01).Flow cytometry was applied to deter-mine the effect of miR-3910 on cell cycle progression,revealing accelerated cell cycle progression,a re-duced proportion of G0/G,phase cells(P＜0.01),and a significant increase in S-phase cells(P＜0.05).Integrated bioinformatics analysis and high-throughput transcriptome sequencing predicted key molecular changes in SH-SY5Y cells following miR-3910 overexpression.Transcriptome sequencing and bioinformatics analysis identified six NB-related genes:EIF3CL(EIF3C),RNF103-CHMP3(VPS24),SULT1A4(SULT1A4),CORO7-PAM16(CORO7),H4C12(Histone H4),and TBC1D3(TBC1D3A/B/C)(aliases sourced from the GeneCards database).qRT-PCR and Western blotting(WB)results are consistency with sequencing results(P＜0.01).In conclusion,miR-3910 overexpression significantly promotes SH-SY5Y cell proliferation,migration,and cell cycle progression,while uncovering a series of potential key target molecules.These findings provide new insights into the pathogenesis of NB and offer a theoretical foundation and potential intervention targets for molecular-targeted therapy in NB.

OBJECTIVE: To explore the efficacy and safety of Juan Bi Pill (JBP) in treatment of active rheumatoid arthritis (RA). METHODS: From February 2017 to May 2018, 115 participants from 4 centers were randomly divided into JBP group (57 cases) and placebo group (58 cases) in a 1:1 ratio using a random number table method. Participants received a dose of JBP (4 g, twice a day, orally) combined with methotrexate (MTX, 10 mg per week) or placebo (4 g, twice a day, orally) combined with MTX for 12 weeks. Participants were required with follow-up visits at 24 and 48 weeks, attending 7 assessment visits. Participants were undergo disease activity assessment 7 times (at baseline and 2, 4, 8, 12, 24, 48 weeks) and safety assessments 6 times (at baseline and 4, 8, 12, 24, 48 weeks). The primary endpoint was 28-joint Disease Activity Score (DAS28-ESR and DAS28-CRP). The secondary endpoints included American College of Rheumatology (ACR) criteria for 20% and 50% improvement (ACR20/50), Health Assessment Questionnaire Disability Index (HAQ-DI), clinical disease activity index (CDAI), visual analog scale (VAS), Short Form-36 (SF-36) score, Medial Outcomes Study (MOS) sleep scale score, serum erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), tender joint count, swollen joint count, and morning stiffness. The adverse reactions were observed during the treatment. RESULTS: After 12 weeks of treatment, DAS28-ESR and DAS28-CRP scores in both groups were lower than before treatment (both P<0.01), while the remission rate of DAS28-ESR and DAS28-CRP and low disease activity of JBP group were higher than those in the placebo group (both P<0.01). JBP demonstrated better efficacy on ACR20 and ACR50 compliance rate at 12 and 48 weeks comparing to placebo (all P<0.05). The CDAI and HAQ-DI score, pain VAS and global VAS change of RA patients and physicians, the serum ESR and CRP levels, and the number of tenderness and swelling joints were lower than before treatment at 4, 8, 12, 24, 48 weeks in both groups (P<0.05 or P<0.01), while the reduction of above indices in the JBP group was more obvious than those in the placebo group at 12 weeks (ESR and CRP, both P<0.05) or at 12 and 48 weeks (all P<0.01). There was no difference in adverse reactions between the 2 groups during treatment (P=0.75). CONCLUSION JBP combined with MTX could effectively reduce disease activity in patients with RA in active stage, reduce the symptoms of arthritis, and improve the quality of life, while ensuring safety, reliability, and fewer adverse effects. (Trial Registration: ClinicalTrials.gov, No. NCT02885597).
Humans ; Arthritis, Rheumatoid/drug therapy* ; Methotrexate/adverse effects* ; Female ; Double-Blind Method ; Male ; Middle Aged ; Treatment Outcome ; Drugs, Chinese Herbal/adverse effects* ; Drug Therapy, Combination ; Adult ; Antirheumatic Agents/adverse effects* ; Aged

Objective:This study aims to evaluate the clinical utility of total IgE （tIgE） and specific IgE （sIgE） levels in nasal secretions for diagnosing allergic rhinitis. The investigation is enhanced through an improved method of nasal secretion collection and advanced quantum dot immunofluorescence detection technology. Methods:A total of 88 subjects were enrolled in this study, and demographic data and clinical characteristics were collected through standardized questionnaires. Anterior rhinoscope was used to check the local condition of the nasal cavity. Each participant underwent skin prick test（SPT）. The total IgE（tIgE） and sIgE in nasal secretions were quantitatively analyzed by improved nasal secretion collection strategy and quantum dot immunofluorescence method, and the correlation between them and clinical symptoms and signs was discussed. The receiver operating characteristic curve（ROC） was used to calculate the optimum threshold and detection efficiency of total IgE and sIgE in nasal secretions. Results:The improved method successfully collected nasal secretions from all subjects. Based on SPT results, participants were categorized into three groups: normal control （20 cases）, non-allergic rhinitis （22 cases）, and allergic rhinitis （46 cases）. Analysis showed that both tIgE and sIgE levels in nasal secretions correlated with nasal symptoms and signs. A tIgE level of ≥9.42 IU/mL was identified as a cut-off for allergic rhinitis diagnosis, demonstrating an 85.37% agreement with SPT results. Furthermore, cut-off values for house dust mite sIgE （≥0.34 IU/mL） and dermatophagoides Farinae sIgE （≥0.41 IU/mL） yielded a diagnostic agreement of 97.56% with SPT. Notably, two patients in the non-allergic rhinitis group tested negative for SPT but positive for dust mite sIgE in nasal secretions and exhibited positive results in the nasal provocation test, indicating potential local allergic rhinitis. Conclusion:The assessment of tIgE and mite-specific IgE levels in nasal secretions presents a rapid, reliable, and non-invasive approach for diagnosing allergic rhinitis, particularly in cases of local allergic rhinitis.
Humans ; Immunoglobulin E/analysis* ; Quantum Dots ; Male ; Female ; Adult ; Young Adult ; Middle Aged ; Rhinitis, Allergic/immunology* ; Adolescent ; Fluorescent Antibody Technique/methods* ; Case-Control Studies ; Nasal Mucosa/immunology*

Eightly-four novel thioheterocyclic nucleoside derivatives were designed, synthesized, and evaluated for antitumor activity in vitro and in vivo. Most of the compounds inhibited the growth of HCT116 and HeLa cancer cells in vitro, among them 33a and 36b exhibited potent activity against HCT116 cells (IC₅₀ = 0.27 and 0.49 μmol/L, respectively). Both compounds 33a and 36b inhibited cell metastasis, arrested the cell cycle in the G₂/M phase, and induced apoptosis in vitro. Mechanistic studies revealed that 33a and 36b increased ROS levels, led to DNA damage, ER stress, and mitochondrial dysfunction, and inhibited autophagy in HCT116 cells. Biological information analysis, RNA-sequencing, Gene Set Enrichment Analysis (GSEA), drug affinity responsive target stability (DARTS) assay, cellular thermal shift assay (CETSA), and SPR experiments identified that compounds 33a and 36b showed antitumor activity by suppressing the c-MYC pathway. c-MYC silencing assays indicated that c-MYC proteins participated in 33a-mediated anticancer activities in HCT116 cells. More importantly, compound 33a presented favorable pharmacokinetic properties in mice (T _1/2 = 6.8 h) and showed significant antitumor efficacy in vivo without obvious toxicity, showing promising potential for further clinical development.