Mutations in the cyclin-dependent kinase-like 5 gene (CDKL5) cause a severe neurodevelopmental disorder, yet the impact of truncating mutations remains unclear. Here, we introduce the Cdkl5^492stop mouse model, mimicking C-terminal truncating mutations in patients. 492stop/Y mice exhibit altered dendritic spine morphology and spontaneous seizure-like behaviors, alongside other behavioral deficits. After creating cell lines with various Cdkl5 truncating mutations, we found that these mutations are regulated by the nonsense-mediated RNA decay pathway. Most truncating mutations result in CDKL5 protein loss, leading to multiple disease phenotypes, and offering new insights into the pathogenesis of CDKL5 disorder.
Animals ; Disease Models, Animal ; Mice ; Protein Serine-Threonine Kinases/deficiency* ; Mutation/genetics* ; Epileptic Syndromes/genetics* ; Humans ; Dendritic Spines/pathology* ; Spasms, Infantile/genetics* ; Male ; Seizures/genetics* ; Mice, Inbred C57BL

High mobility group box 1 (HMGB1), when released extracellularly, plays a pivotal role in the development of spinal cord synapses and exacerbates autoimmune diseases within the central nervous system. In experimental autoimmune encephalomyelitis (EAE), a condition that models multiple sclerosis, the levels of extracellular HMGB1 and interleukin-33 (IL-33) have been found to be inversely correlated. However, the mechanism by which IL-33 deficiency enhances HMGB1 release during EAE remains elusive. Our study elucidates a potential signaling pathway whereby the absence of IL-33 leads to increased binding of P300/CBP-associated factor with HMGB1 in the nuclei of astrocytes, upregulating HMGB1 acetylation and promoting its release from astrocyte nuclei in the spinal cord of EAE mice. Conversely, the addition of IL-33 counteracts the TNF-α-induced increase in HMGB1 and acetylated HMGB1 levels in primary astrocytes. These findings underscore the potential of IL-33-associated signaling pathways as a therapeutic target for EAE treatment.
Animals ; Encephalomyelitis, Autoimmune, Experimental/metabolism* ; Astrocytes/metabolism* ; Interleukin-33/metabolism* ; HMGB1 Protein/metabolism* ; Acetylation ; Mice, Knockout ; Mice, Inbred C57BL ; p300-CBP Transcription Factors/metabolism* ; Mice ; Spinal Cord/metabolism* ; Cells, Cultured ; Female ; Signal Transduction

Targeted covalent inhibitors, primarily targeting cysteine residues, have attracted great attention as potential drug candidates due to good potency and prolonged duration of action. However, their discovery is challenging. In this research, a database-assisted liquid chromatography-tandem mass spectrometry (LC-MS/MS) strategy was developed to quickly discover potential cysteine-targeting compounds. First, compounds with potential reactive groups were selected and incubated with N-acetyl-cysteine in microsomes. And the precursor ions of possible cysteine-adducts were predicted based on covalent binding mechanisms to establish in-house database. Second, substrate-independent product ions produced from N-acetyl-cysteine moiety were selected. Third, multiple reaction monitoring scan was conducted to achieve sensitive screening for cysteine-targeting compounds. This strategy showed broad applicability, and covalent compounds with diverse structures were screened out, offering structural resources for covalent inhibitors development. Moreover, the screened compounds, norketamine and hydroxynorketamine, could modify synaptic transmission-related proteins in vivo, indicating their potential as covalent inhibitors. This experimental-based screening strategy provides a quick and reliable guidance for the design and discovery of covalent inhibitors.

OBJECTIVE: Poxviruses are zoonotic pathogens that infect humans, mammals, vertebrates, and arthropods. However, the specific role of ticks in transmission and evolution of these viruses remains unclear. METHODS: Transcriptomic and metatranscriptomic raw data from 329 sampling pools of seven tick species across five continents were mined to assess the diversity and abundance of poxviruses. Chordopoxviral sequences were assembled and subjected to phylogenetic analysis to trace the origins of the unblasted fragments within these sequences. RESULTS: Fifty-eight poxvirus species, representing two subfamilies and 20 genera, were identified, with 212 poxviral sequences assembled. A substantial proportion of AT-rich fragments were detected in the assembled poxviral genomes. These genomic sequences contained fragments originating from rodents, archaea, and arthropods. CONCLUSION Our findings indicate that ticks play a significant role in the transmission and evolution of poxviruses. These viruses demonstrate the capacity to modulate virulence and adaptability through horizontal gene transfer, gene recombination, and gene mutations, thereby promoting co-existence and co-evolution with their hosts. This study advances understanding of the ecological dynamics of poxvirus transmission and evolution and highlights the potential role of ticks as vectors and vessels in these processes.
Animals ; Poxviridae/physiology* ; Ticks/virology* ; Phylogeny ; Transcriptome ; Evolution, Molecular ; Poxviridae Infections/virology* ; Genome, Viral

Objective To evaluate the predictive value of dose-surface histogram(DSH)for radiation proctitis(RP)in prostate cancer(PCa)patients undergoing radiotherapy.Methods This prospective randomized controlled clinical trial included 380 PCa patients who underwent image-guided radiotherapy in the First Affiliated Hospital of Hebei Northern University from January 2018 to January 2023.Patients were randomly divided into observation group(n=200)and control group(n=180).The rectal dose distribution of patients in the two groups was evaluated by using DSH and dose-volume histogram(DVH),respectively.The receiver operating characteristic(ROC)curve was utilized to evaluate the predictive value of DSH for acute RP,with DVH serving as a reference.Results The difference was not statistically significant in clinical information such as age,KPS score,and body mass index(BMI)between the observation and control groups(P＞0.05),as well as in acute RP incidence(P＞0.05).There were significant differences in S40 and V40,S50 and V50,S60 and V60,S70 and V70,and S78 and V78 between the two groups(P＜0.05).S40,S50,V40,and V50 showed low efficacy(P＜0.001)in predicting acute RP at each level,with AUC≤0.700.S60 and V60 showed moderate efficacy(P＜0.001)in predicting acute RP at each level,with AUC 0.700-0.900.S70,S78,V70 and V78 showed high efficacy(P＜0.001)in predicting acute RP at each level,with AUC＞0.900.Conclusions The predictive value of DSH for rectal toxicity in patients with PCa is basically consistent with that of DVH.It is expected to become a novel and valuable tool for evaluating radiotherapy plans in the future.

Objective To analyze the basic conditions and pathological characteristics of the samples in the Human Brain Bank of Hebei Medical University,which were pathologically diagnosed as cerebral amyloid angiopathy,and to provide reference for the research of related diseases.Methods The basic data of gender,age,apolipoprotein E genotype,pathological classification of cerebral amyloid angiopathy,Alzheimer's disease-related pathological change score,comorbidities and other pathological information were analyzed.Results Up to October 2024,twenty samples were confirmed by pathological diagnosis,with a male to female ratio of 3:1 and an average age of(80.90±8.08)years.Involve three kinds of apolipoprotein E subtype,5 kinds of genotypes(ε2/ε3 xε2/ε4、ε3/ε3 xε3/ε4、ε4/ε4);There were 2 pathologic types,including 6 cases of type 1 and 14 cases of type 2.The pathological grade included 3 grades.The severity grade and subtype classification of cerebral amyloid vascular disease were correlated with the degree of pathological changes of Alzheimer's disease.Cerebral amyloid angiopathy samples could coexist with other degenerative diseases with high comorbidity.Conclusion The incidence of cerebral amyloid angiopathy is higher in the aged samples collected based on Brain Bank,which coexists with conditions such as Alzheimer's disease and microbleeds,etc.It provides more detailed pathological diagnosis basis for further scientific research sharing of samples.

Objective To discuss the application of 3D printing technology in the adjuvant treatment of complex Stanford type B aortic dissection(SBAD)and abdominal aortic aneurysm.Methods The clinical data of 64 patients with complex SBAD and 64 patients with abdominal aortic aneurysm,who were admitted to the Mianyang No.404 Hospital of China from January 2022 to January 2024,were retrospectively analyzed.Of the 64 patients with complex SBAD,33 received preoperative 3D printing adjuvant treatment(observation group Ⅰ)and 31 received preoperative routine examination(control group Ⅰ).Of the 64 patients with abdominal aortic aneurysm,32 received preoperative 3D printing adjuvant treatment(observation group Ⅱ)and 32 received preoperative routine examination(control group Ⅱ).The changes in left-right diameter(LR)and anterior-posterior diameter(AP)of anatomical structure in observation group Ⅰ and observation group Ⅱ were analyzed.The perioperative situations were compared between observation group Ⅰ and control group Ⅰ,as well as between observation groupⅡ and control group Ⅱ.Results In patients with complex SBAD,LR of descending aorta diaphragm in S2(STL model)was significantly higher than that in S1(CTA image)and S3(plastic model,P＜0.05),and AP of descending aorta diaphragm in S2 was higher than that in S3(P＜0.05).LR of brachiocephalic trunk in S3 was significantly lower than that in S1 and S2(P＜0.05),AP of brachiocephalic trunk in S3 stage was significantly higher than that in S1 and S2(P＜0.05),and AP of brachiocephalic trunk in S2 was higher than that in S1(P＜0.05).LR of left common carotid artery in S3 was significantly higher than that in S1 and S2(P＜0.05),LR of left common carotid artery in S2 was higher than that in S1(P＜0.05),and AP of left common carotid artery in S3 was lower than that in S1(P＜0.05).LR and AP of left subclavian artery in S3 were significantly higher than those in S1 and S2(P＜0.05).In patients with abdominal aortic aneurysm,LR and AP of tumor neck in S3 were significantly higher than those in S1(P＜0.05),and AP of aneurysm neck in S3 was significantly higher than that in S2(P＜0.05).LR and AP of aneurysm in S3 and S2 were significantly higher than those in S1(P＜0.05),and LR and AP of aneurysm in S3 were significantly higher than those in S2(P＜0.05).LR of abdominal aortic bifurcation in S3 and S2 was significantly higher than that in S1(P＜0.05),LR of abdominal aortic bifurcation in S3 was significantly higher than that in S2(P＜0.05),and AP of abdominal aortic bifurcation in S3 was significantly lower than that in S1(P＜0.05).AP of left common iliac artery in S3 was significantly lower than that in S1(P＜0.05).In the observation group Ⅰ,the operation time,endovascular operation time and length of hospital stay were significantly shorter than those in the control group Ⅰ(P＜0.05),and the intraoperative blood loss and used dosage of contrast agent were lower than those in the control group Ⅰ(P＜0.05).In observation group Ⅱ,the operation time,endovascular operation time and length of hospital stay were significantly shorter than those in the control group Ⅱ(P＜0.05),and the intraoperative blood loss and used dosage of contrast agent were lower than those in the control group Ⅱ(P＜0.05).In patients with complex SBAD or abdominal aortic aneurysm,there was no internal leakage or stent displacement at 6 months after surgery.Conclusion Adjuvant treatment with 3D printing technology is helpful for improving anatomical structure measurement of lesion sites in patients with complex SBAD and abdominal aortic aneurysm.Preoperative 3D plastic model preview surgery is helpful for shortening the operation time and length of hospital stay and reducing the used dosage of contrast agent without affecting surgical treatment effect.

Objective To explore the impact of interactions among serum CD19+,interleukin-4(IL-4),and Epstein-Barr virus(EBV)DNA load on the occurrence of pediatric infectious mononu-cleosis(IM).Methods A total of 100 IM pediatric patients were enrolled as study group,and 200 healthy pediatric controls were recruited during the same period.Baseline characteristics,EBV-DNA load,CD19+levels,and IL-4 expression were compared between the two groups.A multivariate Lo-gistic regression model was used to analyze the influencing factors of IM.The interactions between EBV-DNA load and CD19+,IL-4 were analyzed based on an additive model.Receiver operating characteristic(ROC)curves were plotted to assess the diagnostic performance of EBV-DNA load,CD19+,IL-4 alone and their combination for IM.Results Significant differences were observed be-tween the two groups in terms of EBV-DNA load,CD19+levels,IL-4 expression,neutrophil-to-lym-phocyte ratio(NLR),monocyte-to-lymphocyte ratio(MLR),red cell distribution width(RDW),atypical lymphocyte ratio,and VCA-IgM positivity(P＜0.05).Multivariate Logistic regression anal-ysis revealed that EBV-DNA load,CD19+,IL-4,RDW,atypical lymphocyte ratio,and VCA-IgM positivity were independent influencing factors for IM occurrence(P＜0.05).Additive interaction analysis showed that when EBV-DNA load and IL-4 were simultaneously exposed,the relative excess risk due to interaction(RERI)was 63.888,the attributable proportion due to interaction(API)was 77.312,and the synergy index(S)was 4.532;when EBV-DNA load and CD19+were simul-taneously exposed,RERI was 2.655,API was 16.773,and S was 1.210.ROC curve analysis indi-cated that the area under the curve(AUC)for the combined diagnosis of IM using EBV-DNA load,CD19+,and IL-4 was 0.945,which was superior to that of single indicator and dual combination.Conclusion Serum CD19+,IL-4,and EBV-DNA load exhibit additive interactions in the occur-rence of IM,and simultaneous exposure increases the risk of IM.Combined detection of these bio-markers enhances the diagnostic performance for IM,providing a reference for clinical diagnosis and treatment.

This study aims to establish the S_(180) tumor-bearing mice model, and to investigate the influence of Shenqi Erpi Granules(SQEPG) on immune function, as well as the drug's tumor-suppressive effect and mechanism. SPF grade KM mice(half male and half female) were randomly divided into 6 groups: a control group, a model group, a cyclophosphamide group(50 mg·kg~(-1)), as well as SQEPG groups in low-, medium-, and high-dose(5.25, 10.5, 21 g·kg~(-1)). The control group and the model group were given distilled water, and the other 4 groups were given the corresponding drugs by gavage. The administration continued for 10 days before the mice were sacrificed. The antitumor and immune regulation effects of SQEPG were evaluated. The effect of SQEPG on delayed type hypersensitivity reaction(DTH), carbon clearance index, and serum hemolysin antibody level was observed to reflect the effect on the immune function of tumor-bearing mice. Tumor weight was recorded to calculate the tumor suppression rate and the immune organ index. Hematoxylin-eosin(HE) staining was used to detect morphological changes in tumor tissues. Flow cytometry was employed to detect the percentage of CD4~+ and CD8~+ T-cells in the spleen tissues and the tumor tissue apoptosis levels. Immunohistochemistry was conducted to detect the KI67 protein expression level of tumor tissues. ELISA resorted to the detection of the following expression levels in tumor tissues: tumor necrosis factor-α(TNF-α), interleukin-2(IL-2), interferon-γ(IFN-γ). Western blot was performed to detect the expression levels of caspase-3, B-cell lymphoma-2(Bcl-2), Bcl-2-associated X protein(Bax), cyclin-dependent kinases 4(CDK4), G_1/S-specific cyclin D1(cyclin D1), and vascular endothelial growth factor A(VEGFA). The results showed that, compared with the model group, the SQEPG could increase the swelling of the auricle of the tumor-bearing mice; significantly increase the phagocytic index of carbon granule contour(P<0.05 or P<0.01), and the middle dose of SQEPG could significantly increase the antibody level of hemolysin(P<0.05); different doses of SQEPG significantly inhibit the growth of the tumor, and decrease the mass of the tumor tissues(P<0.05 or P<0.01); the low dose of SQEPG significantly decreased spleen index(P<0.05), low and high doses of SQEPG increased thymus index, while medium doses of SQEPG decreased thymus index. High doses of SQEPG significantly elevated the levels of CD4~+ and CD8~+ T-cells in the spleens of the homozygous mice(P<0.01 or P<0.001), and increased the apoptosis rate of the cells of the tumor tissues(P<0.05); Meanwhile, high-dose SQEPG elevated the levels of immunity factors such as IL-2, IFN-γ and TNF-α in the serum of tumor-bearing mice(P<0.01); medium-and high-dose SQEPG significantly lowered the rate of positive expression of KI67 protein in tumor tissues(P<0.01). Compared with the model group, high-dose SQEPG significantly up-regulated the expression of caspase-3 and Bax proteins in tumor tissues(P<0.05), and significantly down-regulated the expression of CDK4, cyclin D1, and VEGFA proteins(P<0.05 or P<0.01). In conclusion, SQEPG has the effect of improving immune function and inhibiting tumor growth in tumor-bearing mice. Its mechanism of tumor-suppressive effects may be related to apoptosis promotion, cell cycle progression block, and tumor cell proliferation inhibition.
Animals ; Mice ; Drugs, Chinese Herbal/pharmacology* ; Male ; Female ; Apoptosis/drug effects* ; Sarcoma 180/genetics* ; Humans

OBJECTIVE: To analyze the molecular genetic spectrum of children with acute myeloid leukemia (AML), and explore its correlation with clinical characteristics and prognosis. METHODS: The clinical and molecular genetic data of 116 children with newly diagnosed AML in Wuhan Children's Hospital from September 2015 to August 2022 were retrospectively analyzed. The Fisher's exact test was used to analyze the correlation of gene mutations with clinical features, and Kaplan-Meier curve was used to analyze the influences of gene mutations on the prognosis. RESULTS: NRAS (22%), KRAS (14.9%), and KIT (14.7%) mutations were the most common genetic abnormalities in 116 children with AML. Children with KIT, CEBPA and GATA2 mutations showed a higher median onset-age than those without mutations (all P < 0.05). Children with FLT3-ITD mutation exhibited a higher white blood cell count at initial diagnosis compared to those without mutations (P < 0.05). Children with ASXL2 mutation had lower platelet count and hemoglobin at initial diagnosis than those without mutations (both P < 0.05). KIT mutations were often co-occurred with t(8;21)(q22;q22). There was no significant relationship between gene mutation and minimal residual disease (MRD) remission rate after the first and second induction therapy (P >0.05). KIT and NRAS mutations were not associated with prognosis significantly (P >0.05). The overall survival (OS) rates of children with CEBPA and FLT3-ITD mutations were superior to those without mutations, but the differences were not statistically significant (P >0.05). The 3-year OS rate of 61 children treated by allogeneic hematopoietic stem cell transplantation was 89.8%, which was significantly higher than 55.2% of those only treated by chemotherapy (P < 0.001). CONCLUSIONS Gene mutations are common in children with AML, and next-generation sequencing can significantly improve the detection rate of gene mutations, which can guide the risk stratification therapy. In addition, FLT3-ITD and KIT mutations may no longer be poor prognostic factors.
Humans ; Leukemia, Myeloid, Acute/genetics* ; Mutation ; Prognosis ; Retrospective Studies ; fms-Like Tyrosine Kinase 3/genetics* ; Child ; Proto-Oncogene Proteins c-kit/genetics* ; Male ; Female ; CCAAT-Enhancer-Binding Proteins/genetics* ; Membrane Proteins/genetics* ; Child, Preschool ; Adolescent ; GATA2 Transcription Factor/genetics* ; GTP Phosphohydrolases/genetics* ; Proto-Oncogene Proteins p21(ras)/genetics*