ObjectiveTobacco-related diseases remain one of the leading preventable public health challenges worldwide and are among the primary causes of premature death. In recent years, accumulating evidence has supported the classification of nicotine addiction as a chronic brain disease, profoundly affecting both brain structure and function. Despite the urgency, effective diagnostic methods for smoking addiction remain lacking, posing significant challenges for early intervention and treatment. To address this issue and gain deeper insights into the neural mechanisms underlying nicotine dependence, this study proposes a novel graph neural network framework, termed TI-GNN. This model leverages functional magnetic resonance imaging (fMRI) data to identify complex and subtle abnormalities in brain connectivity patterns associated with smoking addiction. MethodsThe study utilizes fMRI data to construct functional connectivity matrices that represent interaction patterns among brain regions. These matrices are interpreted as graphs, where brain regions are nodes and the strength of functional connectivity between them serves as edges. The proposed TI-GNN model integrates a Transformer module to effectively capture global interactions across the entire brain network, enabling a comprehensive understanding of high-level connectivity patterns. Additionally, a spatial attention mechanism is employed to selectively focus on informative inter-regional connections while filtering out irrelevant or noisy features. This design enhances the model’s ability to learn meaningful neural representations crucial for classification tasks. A key innovation of TI-GNN lies in its built-in causal interpretation module, which aims to infer directional and potentially causal relationships among brain regions. This not only improves predictive performance but also enhances model interpretability—an essential attribute for clinical applications. The identification of causal links provides valuable insights into the neuropathological basis of addiction and contributes to the development of biologically plausible and trustworthy diagnostic tools. ResultsExperimental results demonstrate that the TI-GNN model achieves superior classification performance on the smoking addiction dataset, outperforming several state-of-the-art baseline models. Specifically, TI-GNN attains an accuracy of 0.91, an F1-score of 0.91, and a Matthews correlation coefficient (MCC) of 0.83, indicating strong robustness and reliability. Beyond performance metrics, TI-GNN identifies critical abnormal connectivity patterns in several brain regions implicated in addiction. Notably, it highlights dysregulations in the amygdala and the anterior cingulate cortex, consistent with prior clinical and neuroimaging findings. These regions are well known for their roles in emotional regulation, reward processing, and impulse control—functions that are frequently disrupted in nicotine dependence. ConclusionThe TI-GNN framework offers a powerful and interpretable tool for the objective diagnosis of smoking addiction. By integrating advanced graph learning techniques with causal inference capabilities, the model not only achieves high diagnostic accuracy but also elucidates the neurobiological underpinnings of addiction. The identification of specific abnormal brain networks and their causal interactions deepens our understanding of addiction pathophysiology and lays the groundwork for developing targeted intervention strategies and personalized treatment approaches in the future.

Bombesin receptor subtype-3 (BRS3) is an orphan G protein-coupled receptor (GPCR) that plays critical roles in energy homeostasis, glucose metabolism, and insulin secretion. Recent structural studies have elucidated BRS3 signaling mechanisms using synthetic ligands, including BA1 and MK-5046. However, the molecular basis of BRS3 activation by bioactive natural compounds and their derivatives, particularly those derived from traditional Chinese medicine, remains unclear. Here, we present high-resolution cryogenic electron microscopy (cryo-EM) structures of the human BRS3-G_q complex in both unliganded and active states bound by two herb-derived compounds (DSO-5a and oridonin), at resolutions of 2.9, 2.8, and 2.9 Å, respectively. These structures display distinct ligand recognition patterns between DSO-5a and oridonin. Although both compounds bind to the orthosteric pocket, they differentially engage the interaction network of BRS3, as demonstrated by mutagenesis studies assessing calcium mobilization and inositol phosphate 1 (IP1) accumulation. These findings enhance our understanding of BRS3 activation and provide valuable insights into the development of small-molecule BRS3 modulators with therapeutic potential.

Osteoporosis is a prevalent skeletal condition characterized by reduced bone mass and strength, leading to increased fragility. Buqi-Tongluo (BQTL) decoction, a traditional Chinese medicine (TCM) prescription, has yet to be fully evaluated for its potential in treating bone diseases such as osteoporosis. To investigate the mechanism by which BQTL decoction inhibits osteoclast differentiation in vitro and validate these findings through in vivo experiments. We employed MTS assays to assess the potential proliferative or toxic effects of BQTL on bone marrow macrophages (BMMs) at various concentrations. TRAcP experiments were conducted to examine BQTL's impact on osteoclast differentiation. RT-PCR and Western blot analyses were utilized to evaluate the relative expression levels of osteoclast-specific genes and proteins under BQTL stimulation. Finally, in vivo experiments were performed using an osteoporosis model to further validate the in vitro findings. This study revealed that BQTL suppressed receptor activator of NF-κB ligand (RANKL)-induced osteoclastogenesis and osteoclast resorption activity in vitro in a dose-dependent manner without observable cytotoxicity. The inhibitory effects of BQTL on osteoclast formation and function were attributed to the downregulation of NFATc1 and c-fos activity, primarily through attenuation of the MAPK, NF-κB, and Calcineurin signaling pathways. BQTL's inhibitory capacity was further examined in vivo using an ovariectomized (OVX) rat model, demonstrating a strong protective effect against bone loss. BQTL may serve as an effective therapeutic TCM for the treatment of postmenopausal osteoporosis and the alleviation of bone loss induced by estrogen deficiency and related conditions.
Animals ; NFATC Transcription Factors/genetics* ; Drugs, Chinese Herbal/pharmacology* ; Ovariectomy ; Osteoclasts/metabolism* ; Female ; Osteogenesis/drug effects* ; Rats, Sprague-Dawley ; Rats ; NF-kappa B/genetics* ; Osteoporosis/genetics* ; Signal Transduction/drug effects* ; Bone Resorption/genetics* ; Cell Differentiation/drug effects* ; Humans ; RANK Ligand/metabolism* ; Mitogen-Activated Protein Kinases/genetics* ; Transcription Factors

BACKGROUND:The specific molecular mechanism of the transformation from normal healthy people to acute cervical spondylotic radiculopathy has not been clear,which needs to be further studied. OBJECTIVE:To investigate the differential expression of serum proteomics between normal healthy people and patients with acute cervical spondylotic radiculopathy,and to find and identify potential specific serum markers between them. METHODS:The serum samples of eight patients with acute cervical spondylotic radiculopathy and eight normal healthy people were collected,and the proteomic screening and analysis were performed by tandem mass tag combined with liquid chromatography-tandem mass spectrometry technology,in order to explore and identify serum proteins differentially expressed in patients with acute cervical spondylotic radiculopathy. RESULTS AND CONCLUSION:A total of 183 significantly differential proteins were screened by tandem mass tag technology,and 11 significantly differential proteins were identified(P＜0.05).Compared with normal healthy people,three differential proteins were significantly up-regulated,including human leukocyte antigen-A,secretoglobin family 1a member 1,and protein 4-hydroxyphenylpyruvate dioxygenase,and seven differential proteins were significantly down-regulated,such as immunoglobulin heavy constant gamma 3,skin factor,and myosin light chain 3,in patients with acute cervical spondylotic radiculopathy.Gene ontology enrichment analysis showed that these differential proteins participated in antigen binding,immunoglobulin receptor binding and other molecular functions.Protein-protein interaction analysis showed that among the common differential proteins between normal healthy people and patients with acute cervical spondylotic radiculopathy,HLA-A,HPD,PSMA3,DMKN,SCGB1A1,and MYL3 were located at the nodes of the functional network,and were closely related to the systems of body immunity,cellular inflammatory response,energy metabolism,and mechanical pressure.The significantly differential proteins HLA-A,HPD and MYL3 were verified by western blot,and the results were consistent with those of proteomics.To conclude,tandem mass tag combined with liquid chromatography-tandem mass spectrometry technology can be used to find the differentially expressed proteins in serum between normal healthy people and patients with acute cervical spondylotic radiculopathy.It is preliminarily believed that HLA-A,HPD and MYL3 may be specific serum markers of acute cervical spondylotic radiculopathy,providing a new direction for further research on its pathogenesis.

Objective To observe the value of electron density map(EDM)from spectral CT combined with CT features in differentiating acute and chronic osteoporotic vertebral fractures(OVF).Methods Thoracic and/or lumbar spectral CT data of 48 patients with acute complicated chronic OVF were retrospectively analyzed.Totally 110 fractured vertebrae were enrolled,including 53 vertebrae with acute fractures(acute group)and 57 with chronic fractures(chronic group).The quantitative parameters of spectral CT,including CT values of conventional 120 kVp polyenergetic image(PI,i.e.routine CT images)and 40,70,100 keV virtual monoenergetic images(VMI),effective atomic number(Z-eff)and electron density(ED),as well as routine CT finding were compared between groups,and those being significantly different were included in multivariate logistic regression to screen the independent risk factors for acute OVF and construct a combined model.Receiver operating characteristic(ROC)curves were drawn to evaluate the efficacy of each single independent risk factor and the combination for differentiating acute and chronic OVF.Results Significant differences of all spectral CT quantitative parameters,also of routine CT findings including interruption of vertebral endplate,cortical folds,increased vertebral density,gas within vertebral body and vertebral compression degree were found between groups(all P＜0.05).Logistic regression analysis showed that CTPI(OR=0.855,P=0.005),ED(OR=16.432,P=0.005),cortical folds(OR=0.038,P=0.034)and increased vertebral density(OR=0.025,P=0.013)were all independent risk factors for acute OVF.The area under the curve(AUC)of the above single parameters for identifying acute and chronic OVF was 0.870,0.889,0.879 and 0.866,respectively,all lower than that of the combined model(0.977)(Z=3.47,3.73,2.95,2.71,all P＜0.05).Conclusion Spectral CT EDM combined with CT findings could effectively differentiate acute and chronic OVF.

Background::The goal of the assisted reproductive treatment is to transfer one euploid blastocyst and to help infertile women giving birth one healthy neonate. Some algorithms have been used to assess the ploidy status of embryos derived from couples with normal chromosome, who subjected to preimplantation genetic testing for aneuploidy (PGT-A) treatment. However, it is currently unknown whether artificial intelligence model can be used to assess the euploidy status of blastocyst derived from populations with chromosomal rearrangement.Methods::From February 2020 to May 2021, we collected the whole raw time-lapse videos at multiple focal planes from in vitro cultured embryos, the clinical information of couples, and the comprehensive chromosome screening results of those blastocysts that had received PGT treatment. Initially, we developed a novel deep learning model called the Attentive Multi-Focus Selection Network (AMSNet) to analyze time-lapse videos in real time and predict blastocyst formation. Building upon AMSNet, we integrated additional clinically predictive variables and created a second deep learning model, the Attentive Multi-Focus Video and Clinical Information Fusion Network (AMCFNet), to assess the euploidy status of embryos. The efficacy of the AMCFNet was further tested in embryos with parental chromosomal rearrangements. The receiver operating characteristic curve (ROC) was used to evaluate the superiority of the model. Results::A total of 4112 embryos with complete time-lapse videos were enrolled for the blastocyst formation prediction task, and 1422 qualified blastocysts received PGT-A ( n = 589) or PGT for chromosomal structural rearrangement (PGT-SR, n = 833) were enrolled for the euploidy assessment task in this study. The AMSNet model using seven focal raw time-lapse videos has the best real-time accuracy. The real-time accuracy for AMSNet to predict blastocyst formation reached above 70% on the day 2 of embryo culture, and then increased to 80% on the day 4 of embryo culture. Combing with 4 clinical features of couples, the AUC of AMCFNet with 7 focal points increased to 0.729 in blastocysts derived from couples with chromosomal rearrangement. Conclusion::Integrating seven focal raw time-lapse images of embryos and parental clinical information, AMCFNet model have the capability of assessing euploidy status in blastocysts derived from couples with chromosomal rearrangement.

ObjectiveTo establish a vaginal self-sampling HPV cervical cancer screening model in Hainan Province, to analyze the application of p16 protein detection in HPV positive and non-HPV16 /18 shunt screening. MethodsFrom January 2019 to September 2022, a total of 200 women from the targeted population was randomly selected for vaginal self-sampling HPV typing test to screen cervical cancer using randomized numeric table method, followed by cervical cytology sampling for cytology p16 protein detection. Postoperative pathological examination was used as the gold standard. Multivariate logistic regression analysis was used to analyze the influencing factors of HPV positive detection rate in cervical lesions, and the nomogram model was constructed simultaneously. The receiver operating characteristic(ROC) curve and calibration curve were used for evaluating the accuracy of the nomogram model. Differences in the distribution of self-sampled HPV-positive and HPV infected genotypes were recorded, and the application of p16 protein detection in HPV-positive and non-HPV16/18 shunt screening was analyzed. ResultsAged ≥40 years, BMI ≥28.00 kg·m^-2, number of sexual partners ≥2, frequency of sexual life ≥10 times·month^-1, bleeding from sexual intercourse, and age of first sexual intercourse <22 years were the risk factors for HPV positive of cervical lesions (all P<0.001). The results of ROC curve and calibration curve showed that the area under ROC curve (AUC) was 0.874 (95%CI: 0.823‒0.907, P<0.05), the sensitivity was 0.835, the specificity was 0.847, and the Youden index was 0.672, indicating a good fit of the model. Results of vaginal self-sampling HPV test showed that the positive rate of HPV was 86.50% (173/200). HPV high-risk infection types mainly included HPV16, 18, 31, 33, 35, 39, 45, 52, 56, 58, 59, 68, 73, and 82. Single HPV infection accounted for 95.95% (166/173), 2.89% (5/173) were infected with two types of HPV, and 1.16% (2/173) were infected with three or more types of HPV. Colposcopic pathologic diagnosis was used as the gold standard, and the results showed that the accuracy of p16 protein detection in the diagnosis of cervical cancer was 93.50% (187/200), with a sensitivity of 96.53% (167/173), and a specificity of 74.07% (20/27). The negative and positive predictive value were 76.92% (20/26) and 95.98% (167/174), respectively. The results of shunt screening showed that there were 80 cases infected with HPV16, 79 cases infected with HPV18 and 41 cases of non-HPV16/18, with a sensitivity of 90.91%, 90.32% and 86.67%, a specificity of 71.43%, 64.71% and 72.73%, a negative predictive value of 62.50%, 64.71% and 66.67%, a positive predictive value of 93.75%, 90.32% and 89.66%, and an accuracy of 87.50%, 84.81% and 82.93%, respectively. The specificity and accuracy of p16 positive screening for cervical cancer were significantly higher than that of HPV positive detection, but the false positive rate was significantly lower than that of HPV positive detection. The AUCs of HPV positive, p16 positive and combination of the two detection methods for cervical cancer were 0.603, 0.822 and 0.907, respectively. ConclusionVaginal self-sampling HPV testing is a widely accepted mode for cervical cancer screening. Cervical cytology p16 protein detection is important for self-sampled HPV positive and shunt screening of non-HPV16/18.

Objective To investigate the effect of Anchusa italica Retz.extract on cough variant asthma(CVA)and its mechanism of action.Methods Forty-eight SD rats of each eight were randomly divided into blank group(equal amount of saline),model group(equal amount of saline),control group(250 mg·kg-1 prednisone acetate)and experimental-A1,-A5,-A7 groups(61.1 mg·kg-1 Anchusa italica Retz.extract A1,48.8 mg·kg-1 Anchusa italica Retz.extract A5,201.8 mg·kg-1 Anchusa italica Retz.extract A7),continuous gavage for 28 d;except for the blank group,the other groups were made with ovalbumin-complete Freund's adjuvant.The number of cough times of each rat was observed.The changes in the expression of Toll-like receptor 4(TLR4),myeloid differentiation factor(MyD88)and nuclear factor κB(NF-κB)proteins were observed in the lung tissue.Results The cough times in the blank group,model group,control group and experimental-A1,-A5,-A7 groups were 1.10±0.22,8.33±1.24,3.08±0.65,3.31±0.99,3.08±1.02 and 3.06±0.68;the relative expression levels of TLR4 protein were 0.61±0.01,0.84±0.04,0.66±0.02,0.64±0.04,0.64±0.03 and 0.69±0.02;the relative expression levels of MyD88 protein were 0.54±0.08,0.86±0.06,0.71±0.06,0.65±0.05,0.64±0.08 and 0.70±0.06;the relative expression levels of p-NF-κB protein were 0.48±0.11,0.94±0.06,0.80±0.08,0.68±0.04,0.68±0.06 and 0.78±0.09.Compared with the experimental-A1,-A5,-A7 groups,control group,normal group,the differences were statistically significant in the model group(all P＜0.05).Conclusion The extract of Anchusa Italica Retz.can inhibit the symptoms of CVA rats,and the mechanism may be related to the inhibition of TLR4/MyD88/NF-κB signaling pathway.

Objective To investigate the molecular mechanism of action of isoglycoside-4'-O-apigenin(1 → 2)glucoside(licraside)intervention in alleviating cholestasis.Methods Farnesol X receptor(FXR)was silenced in human HepG2 cells using lentivirus,and bilirubin and α-isothiocyanate(ANIT)were used to induce HepG2 cells to construct a hypercholate-hyperbilirubin model.The effects of licraside on cell viability,biochemical indices contents and the expression level of FXR and its downstream related proteins in the model were investigated.HepG2 cells and FXR-silenced HepG2 cells were divided into normal,model and experimental groups.Bilirubin and probenecid were added to all groups except the normal group,and 80 μmol·L-1 licraside was added to the experimental group.After 24 h of culture,cell viability and the levels of total bile acids(TBA),bilirubin and other biochemical indices were examined in each group;the protein expression levels of FXR and bile salt efflux pump(BSEP)were examined in each group by Western blot assay.Results The cell viability in HepG2 cells normal group,HepG2 cells model group,HepG2 cells experimental group,siFXR-HepG2 cells normal group,siFXR-HepG2 cells model group and siFXR-HepG2 cells experimental group were(100.00±17.15)％,(39.41±2.91)％,(70.79±3.74)％,(81.41±5.12)％,(33.49±2.69)％and(44.08±4.82)％;the levels of TBA were(7.98±5.87),(46.18±10.93),(9.25±7.20),(11.18±3.36),(38.28±5.12)and(34.79±5.39)μmol·L-1;the levels of total bilirubin were(5.21±3.27),(40.29±24.88),(5.21±2.64),(12.00±4.64),(56.36±14.85)and(15.39±5.56)μmol·L-1;the relative expression levels of FXR protein were 1.00±0.10,0.81±0.07,1.11±0.09,0.10±0.02,0.12±0.02 and 0.10±0.04;the relative expression levels of BSEP protein were 1.00±0.17,0.81±0.02,0.88±0.03,0.70±0.09,0.49±0.07 and 0.60±0.10.The differences of the above indexes in the HepG2 cells experimental group compared with the model group were statistically significant(P＜0.001,P＜0.01);except for TBA,the differences of the above indexes in siFXR-HepG2 cells experimental group compared with the model group were statistically significant(P＜0.001,P＜0.01).Conclusion Licraside can effectively reduce the biochemical indices level of hypercholate-hyperbilirubin cell model.This effect is achieved by agonizing of FXR protein expression,increase the efflux of bile acid salt and then reduce the synthesis of bile acids,and eventually achieve the effect of alleviating the cholestasis.

Objective To determine the relationship between methylenetetrahydrofolate reductase(MTHFR)C677T,A1298C and multidrug resistance 1(ABCB1)C3435T gene polymorphisms and the adverse reactions of high-dose methotrexate(HD-MTX)in children with acute lymphoblastic leukemia(ALL).Methods Blood samples of ALL children treated with HD-MTX chemotherapy were collected,and the polymorphic polymorphism of MTHFR C677T,A1298C and ABCB1 C3435T genes were detected by sequencing method.The adverse drug reactions after MTX chemotherapy were evaluated according to the common adverse drug reaction grading criteria.The relationship between MTHFR C677T,A1298C and ABCB1 C3435T gene polymorphisms and HD-MTX adverse drug reactions was analyzed.Results The risk of MTHFR A1298C AC+CC type hepatic injury(grade 2)was higher than AA type[odds ratio(OR)2.350,95％confidence interval(CI)=1.038-5.320,P＜0.05],no correlations were found between MTHFR A1298C and myelosuppression,mucositis,gastrointestinal reaction and acute renal impair(all P＞0.05).ABCB1 C3435T CT type hepatic injury(grade 2)was higher than TT type(OR 5.161,95％CI 1.371-19.424,P＜0.05),ABCB1 C3435T CC+CT type hepatic injury(grade 2)was higher than TT type(OR 4.231,95％CI 1.165-15.362,P＜0.05);no correlations were found between ABCB1 C3435T and myelosuppression,mucositis,gastrointestinal reaction and acute renal impair(all P＞0.05).No correlations wre found between MTHFR C677T and HD-MTX adverse drug reactions(all P＞0.05).Conclusion When treating ALL with HD-MTX,adverse drug reactions can be predicted by detecting MTHFR A1298C and ABCB1 C3435T genotypes,so as to implement more scientific individualized medication.