Based on the ultra performance liquid chromatography-quadrupole Exactive Orbitrap mass spectrometry(UPLC-Q-Exactive Orbitrap-MS) technology, combined with related literature, databases, and reference material information, this study qualitatively analyzed the components of Dayuanyin in the tissue of rats after gavage and employed molecular docking technology to predict the rationality of the mechanism behind the antipyretic effect of the in vivo components in Dayuanyin. A total of 21, 26, 20, 21, 14, and 31 prototype components and 3, 16, 3, 7, 5, and 24 metabolites were identified from the heart, liver, spleen, lung, kidney, and hypothalamus of the rats, respectively, and the binding ability of key components and targets was further verified by molecular docking. The results showed that all components had good binding ability with targets. The established UPLC-Q-Exactive Orbitrap-MS could effectively and quickly identify the Dayuanyin components distributed in tissue and preliminarily identify their metabolites. Many components were identified in the hypothalamus, which suggested that the components delivered to the brain should be focused on in the study on Dayuanyin in the treatment of febrile diseases. The molecular docking technology was used to predict the rationality of the mechanism behind its antipyretic effect, which lays the foundation for the clarification of the material basis and action mechanism of Dayuanyin, the development of new preparations, and the prediction of quality markers.
Animals ; Drugs, Chinese Herbal/administration & dosage* ; Rats ; Molecular Docking Simulation ; Male ; Antipyretics/metabolism* ; Rats, Sprague-Dawley ; Tissue Distribution ; Mass Spectrometry ; Chromatography, High Pressure Liquid ; Hypothalamus/metabolism*

OBJECTIVE: To explore the role of semaphorin 4D (Sema4D) in immunoglobulin A (IgA) -mediated immune abnormalities in B lymphocytes of pediatric Henoch-Schonlein purpura (HSP). METHODS: One hundred HSP children admitted to Hengshui People's Hospital from January 2022 to January 2023 were selected as HSP group, and one hundred healthy children as control group. Sema4D expression was detected, and the relationship between Sema4D expression in children's serum and skin lesions and clinical characteristics of children was analyzed. Sema4D expression on the surface of lymphocytes of HSP children was detected. Different concentrations of human recombinant Sema4D protein was used to stimulate peripheral blood mononuclear cells in HSP children in vitro. The expression level of IgA in the supernatant was detected to verify whether Sema4D mediates immune abnormalities through IgA secreted by B lymphocytes. RESULTS: The Sema4D level in the HSP group was significantly higher than that in the control group (P <0.001). Sema4D level in HSP children with severe, renal involvement, and joint involvement was higher than those with mild to moderate disease, and no renal or joint involvement (all P <0.001). Compared with control group, IgA level, CD8 ⁺ T lymphocyte proportion, and CD19 ⁺ B lymphocyte proportion in the HSP group were significantly higher but CD4 ⁺ T lymphocyte proportion was lower (all P <0.001). The expression levels of Sema4D on the surface of CD4 ⁺ T lymphocytes, CD8 ⁺ T lymphocytes, and CD19 ⁺ B lymphocytes in the HSP group were significantly higher than those in the control group (all P <0.001). With the increase of human recombinant Sema4D protein concentration, the level of IgA expression in HSP children gradually increased (P <0.05). Correlation analysis showed that Sema4D was significantly positively correlated with IgA (r =0.667). CONCLUSION HSP children show high expression of Sema4D, especially on the surface of T and B lymphocytes. The shedding of Sema4D from membrane surface may stimulate B lymphocytes to secrete IgA by binding to CD72, leading to immune abnormalities.
Humans ; IgA Vasculitis/immunology* ; Semaphorins/metabolism* ; B-Lymphocytes/metabolism* ; Immunoglobulin A/immunology* ; Child ; Antigens, CD/metabolism* ; Male ; Female ; Child, Preschool

Objective:To analyze the correlation between sex hormone levels and micropenis after mild hypospadias surgery in children.Methods:A case control study was carried out.The clinical data of 71 children aged 1 to 13 years who underwent mild hypospadias surgery at the First Affiliated Hospital of Xinxiang Medical University from April 2022 to April 2024 were analyzed.Preoperatively, the children were divided into a mild hypospadias group (Group A) and a mild hypospadias with micropenis group (Group B) based on the stretched penile length (SPL).Prolactin, follicle-stimulating hormone (FSH), luteinizing hormone (LH), progesterone, and testosterone (TES) levels of the 2 groups were measured.Multivariate Logistic regression analysis was used to construct a risk prediction model.The discrimination capability of the model was evaluated using the receiver operating characteristic (ROC) curve.SPL and sex hormone levels were measured again 6 months after surgery.The children were divided into a normal penile group (Group AA) and a micropenis group (Group BB) after mild hypospadias according to SPL.Multivariate Logistic regression analysis was used to construct a risk prediction model, which was evaluated using the ROC curve.Results:The levels of FSH, LH and TES in group A before the operation were 3.28(2.02, 4.46) IU/L, 0.53(0.25, 0.79) IU/L and 25.24(17.94, 36.67) ng/dL, respectively, and those in group B were 1.42(1.10, 1.84) IU/L, 0.14(0.09, 0.23) IU/L and 15.73 (12.92, 17.00) ng/dL, respectively.The difference was all statistically significant (all P<0.05).Multivariate Logistic regression analysis showed statistical significance ( OR=0.515, 95% CI: 0.271-0.977; OR=0.035, 95% CI: 0.002-0.542; OR=0.883, 95% CI: 0.796-0.980).The area under the ROC curve (AUC) of the prediction model was 0.906, with a sensitivity of 75.00% and a specificity of 95.74%.The levels of FSH, LH and TES in the postoperative AA group were 2.97 (1.88, 4.28) IU/L, 0.46 (0.23, 0.78) IU/L and 20.92 (17.34, 33.27) ng/dL, respectively.The median levels of FSH, LH and TES in the BB group were 1.52 (1.27, 1.82) IU/L, 0.17 (0.12, 0.26) IU/L and 15.08(11.68, 16.68) ng/dL, respectively.The difference was all statistically significant (all P<0.05).Multivariate Logistic regression analysis showed statistical significance ( OR=0.484, 95% CI: 0.236-0.992; OR=0.061, 95% CI: 0.004-0.939; OR=0.891, 95% CI: 0.795-0.999).The AUC of the prediction model constructed was 0.877, with a sensitivity of 94.12% and a specificity of 68.52%. Conclusions:Lower FSH, LH and TES levels are risk factors for the micropenis after mild hypospadias surgery, and preoperative hormone levels have higher predictive value.

Objective:To analyze the correlation between sex hormone levels and micropenis after mild hypospadias surgery in children.Methods:A case control study was carried out.The clinical data of 71 children aged 1 to 13 years who underwent mild hypospadias surgery at the First Affiliated Hospital of Xinxiang Medical University from April 2022 to April 2024 were analyzed.Preoperatively, the children were divided into a mild hypospadias group (Group A) and a mild hypospadias with micropenis group (Group B) based on the stretched penile length (SPL).Prolactin, follicle-stimulating hormone (FSH), luteinizing hormone (LH), progesterone, and testosterone (TES) levels of the 2 groups were measured.Multivariate Logistic regression analysis was used to construct a risk prediction model.The discrimination capability of the model was evaluated using the receiver operating characteristic (ROC) curve.SPL and sex hormone levels were measured again 6 months after surgery.The children were divided into a normal penile group (Group AA) and a micropenis group (Group BB) after mild hypospadias according to SPL.Multivariate Logistic regression analysis was used to construct a risk prediction model, which was evaluated using the ROC curve.Results:The levels of FSH, LH and TES in group A before the operation were 3.28(2.02, 4.46) IU/L, 0.53(0.25, 0.79) IU/L and 25.24(17.94, 36.67) ng/dL, respectively, and those in group B were 1.42(1.10, 1.84) IU/L, 0.14(0.09, 0.23) IU/L and 15.73 (12.92, 17.00) ng/dL, respectively.The difference was all statistically significant (all P<0.05).Multivariate Logistic regression analysis showed statistical significance ( OR=0.515, 95% CI: 0.271-0.977; OR=0.035, 95% CI: 0.002-0.542; OR=0.883, 95% CI: 0.796-0.980).The area under the ROC curve (AUC) of the prediction model was 0.906, with a sensitivity of 75.00% and a specificity of 95.74%.The levels of FSH, LH and TES in the postoperative AA group were 2.97 (1.88, 4.28) IU/L, 0.46 (0.23, 0.78) IU/L and 20.92 (17.34, 33.27) ng/dL, respectively.The median levels of FSH, LH and TES in the BB group were 1.52 (1.27, 1.82) IU/L, 0.17 (0.12, 0.26) IU/L and 15.08(11.68, 16.68) ng/dL, respectively.The difference was all statistically significant (all P<0.05).Multivariate Logistic regression analysis showed statistical significance ( OR=0.484, 95% CI: 0.236-0.992; OR=0.061, 95% CI: 0.004-0.939; OR=0.891, 95% CI: 0.795-0.999).The AUC of the prediction model constructed was 0.877, with a sensitivity of 94.12% and a specificity of 68.52%. Conclusions:Lower FSH, LH and TES levels are risk factors for the micropenis after mild hypospadias surgery, and preoperative hormone levels have higher predictive value.

Aim To explore the effect of berberine (B E) on RSV infected HEp-2 cells and the related mechanism. Methods HEp-2 cells were infected with RSV and treated with BE. Cell viability was assessed using the CCK-8 assay. Protein expression levels of NLRP3, ASC, caspase-1, PINK1, Parkin, Beclinl, p62, LC3 I,LC3 II,and BNIP3 in HEp-2 cells were detected by Western blot. The secretion level of IL-1 p in HEp-2 cells was measured using ELISA. Apoptosis rate and mitochondrial membrane potential of HEp-2 cells were examined by flow cytometry. Mitochondrial ROS (mtROS) in HEp-2 cells was detected through MitoSOX staining. Colocalization of mitochondria and autophagosomes in HEp-2 cells was investigated using immunofluorescence staining. Cyclosporin A was used for validation experiments. Results BE could significantly improve the activity of RSV-infected HEp-2 cells,reduce the apoptosis rate (P < 0. 05), and decrease the activation level of NLRP3 inflammasomes and IL-lp level (P <0. 05); BE improved mitochondrial function by increasing mitochondrial membrane potential and ATP levels,and reduced mtROS. BE significantly promoted the colocalization of mitochondria-autophagosome in RSV infected cells, inducing PINK1/ Parkin and BNIP3 to mediate mitochondrial autophagy; cyclosporine A aggravated RSV infection. Conclusions BE has protective effects on HEp-2 cells infected by RSV. The mechanism may be related to the inhibitory effect of BE on the production of mtROS and the activation of NLRP3 inflammasomes by inducing PINK1/ Parkin and BNIP3-mediated mitochondrial autophagy.

AIM To establish a UPLC-MS/MS method for the simultaneous content determination of gallic acid,protocatechuic acid,neomangiferin,catechin,caffeic acid,mangiferin,isomangiferin,albiflorin,paeoniflorin,vitexin,liquiritin,scutellarin,baicalin,liquiritigenin,timosaponin BⅡ,quercetin,wogonoside,benzoylpaeoniflorin,isoliquiritigenin,honokiol,magnolol,norarecaidine,arecaidine,arecoline,epicatechin,baicalein,glycyrrhizinate and wogonin in Dayuan Drink.METHODS The analysis was performed on a 35℃thermostatic Syncronis C18 column(100 mm×2.1 mm,1.7 μm),with the mobile phase comprising of 0.1%formic acid-acetonitrile flowing at 0.3 mL/min in a gradient elution manner,and electron spray inoization source was adopted in positive and negative ion scanning with select reaction monitoring mode.RESULTS Twenty-eight constituents showed good linear relationships within their own ranges(R2≥0.991 0),whose average recoveries were 95.60%-103.53%with the RSDs of 0.60%-5.45%.CONCLUSION This rapid,simple,selective,accurate and reliable method can be used for the quality control of Dayuan Drink.

Through a compound induction method, combined with neurobehavioral, macroscopic characterization and objective pathological evaluation indicators, a murine depression model of liver depression transforming into fire syndrome was constructed and confirmed. The model was constructed using a combination of sleep deprivation, light exposure, and alternate-day food deprivation. Evaluation was conducted at three levels: face validity, constructs validity, and predictive validity. The establishment of the liver depression transforming into fire syndrome depression model was further validated through the counterproof of traditional Chinese medicine formulas. In terms of face validity, compared to the control group, mice in the model group exhibited typical depressive symptoms in neurobehavioral assessments; the general observation of the model group mice reveals disheveled and lackluster fur, along with delayed and easily agitated responses. Additionally, there is a substantial increase in water consumption. In the sleep phase detection of mouse, the model group showed a significant increase in the proportion of time spent in the wake phase during sleep, accompanied by a significant decrease in the proportions of time spent in both non-rapid eye movement (NREM) and rapid eye movement (REM) sleep phases. There are significant differences in physiological indicators such as average blood flow velocity, blood flow rate, tongue, urine, and claw color (r values) in the internal carotid artery. Structural validity demonstrated that levels of 5-hydroxytryptamine (5-HT), dopamine (DA), and γ-aminobutyric acid (GABA) in the hippocampus of model mice decreased significantly, while acetylcholine (ACh), tryptophan (Try), and glutamic acid (Glu) levels increased significantly. Treatment with Danzhi Xiaoyao San led to varying degrees of restoration in the aforementioned neurotransmitters. In terms of predictive validity, the antidepressant paroxetine effectively ameliorated depressive behaviors in the model mice, and the classic formula Danzhi Xiaoyao San demonstrated varying degrees of improvement in depression-related indicators. In conclusion, this study has established a novel animal model of liver-stagnation with the fire depression, thereby expanding the repertoire of traditional Chinese medicine depression models. This development contributes to the diversification of melancholic syndrome models in Chinese medicine, offering a broader spectrum of options for scientific exploration, efficacy evaluation, and drug screening in the future prevention and treatment of depression with traditional Chinese medicine. Animal experiments were conducted with the approval and supervision of the Animal Ethics Committee of Jiangxi University of Traditional Chinese Medicine (ethics number: 20230313040).

Objective Viral encephalitis is an infectious disease severely affecting human health.It is caused by a wide variety of viral pathogens,including herpes viruses,flaviviruses,enteroviruses,and other viruses.The laboratory diagnosis of viral encephalitis is a worldwide challenge.Recently,high-throughput sequencing technology has provided new tools for diagnosing central nervous system infections.Thus,In this study,we established a multipathogen detection platform for viral encephalitis based on amplicon sequencing. Methods We designed nine pairs of specific polymerase chain reaction(PCR)primers for the 12 viruses by reviewing the relevant literature.The detection ability of the primers was verified by software simulation and the detection of known positive samples.Amplicon sequencing was used to validate the samples,and consistency was compared with Sanger sequencing. Results The results showed that the target sequences of various pathogens were obtained at a coverage depth level greater than 20×,and the sequence lengths were consistent with the sizes of the predicted amplicons.The sequences were verified using the National Center for Biotechnology Information BLAST,and all results were consistent with the results of Sanger sequencing. Conclusion Amplicon-based high-throughput sequencing technology is feasible as a supplementary method for the pathogenic detection of viral encephalitis.It is also a useful tool for the high-volume screening of clinical samples.

Objective:To investigate the effects of selinexor,a inhibitor of nuclear export protein 1(XPO1)on the proliferation inhibition and apoptosis of Kasumi-1 cells in acute myeloid leukemia(AML).Methods:MTS method was used to detect the inhibitory effect of different concentrations of selinexor on the proliferation of Kasumi-1 cells at different time points.The apoptosis rate and cell cycle changes after treatment with different concentration of selinexor were detected by flow cytometry.Results:Selinexor inhibited the growth of Kasumi-1 cells at different time points in a concentration-dependent manner(r24 h=0.7592,r48 h=0.9456,and r72 h=0.9425).Selinexor inhibited Kasumi-1 cells growth in a time-dependent manner(r=0.9057 in 2.5 μmol/L group,r=0.9897 in 5 μmol/L group and r=0.9994 in 10 μmol/L group).Selinexor could induce apoptosis of Kasumi-1 cells in a dose-dependent manner(r=0.9732),and the apoptosis of Kasumi-1 cells was more obvious with the increase of drug concentration.The proportion of G0/G1 phase was significantly increased and the proportion of S phase was significantly decreased after the treatment of Kasumi-1 cells by selinexor.With the increase of drug concentration,the proportion of Kasumi-1 cells cycle arrest in G0/G1 phase was increased and the cell synthesis was decreased.Conclusion:Selinexor can promote the death of tumor cells by inhibiting Kasumi-1 cells proliferation,inducing apoptosis and blocking cell cycle.