OBJECTIVE To investigate the expression levels of nucleotide-binding oligomerization domain-like recep-tor pyrin domain-associated protein 3(NLRP3)inflammasome pathway and toll-like receptors(TLRs)pathway in serofast patients with syphilis infection,and their correlation with helper T lymphocytes(Th)1/Th2.METHODS A total of 549 patients with syphilis admitted to Tianjin Chest Hospital and Tianjin Medical University General Hospital from Jan.2021 to Dec.2022 were selected.Among them,62 patients who had not received anti-syphilis treatment were assigned to the syphilis group,238 patients whose serum remained positive after anti-syphilis treatment were selected as the fixed group,249 patients whose serum turned negative after anti-syphilis treatment were selected as the negative conversion group,and another 565 healthy people who underwent medical examina-tion in the hospital during the same period were selected as the control group.TLR2,TLR4,nuclear factor(NF)-κB,NLRP3,apoptosis-associated speck-like protein(ASC),cysteinyl aspartate proteolytic enzyme 1(Caspase-1),in-terleukin(IL)-1β mRNA and interferon-γ(IFN-γ),IL-12,IL-13 and IL-10 levels were compared between each group.Pearson's analysis was used to analyze the correlation between the mRNA expression of TLR2,TLR4,NF-κB,NLRP3,ASC,Caspase-1,and IL-1β and the levels of IFN-γ,IL-12,IL-13,and IL-10 in serologically fixed patients with syphilis.RESULTS Compared with the control group,the mRNA expression of TLR2,TLR4,NF-κB,NLRP3,ASC,Caspase-1 and IL-1β was elevated in the negative conversion and the syphilis groups,and the expres-sion of the above indicators was decreased in the fixed group(P＜0.05).Compared with negative conversion group,mR-NA expressions of TLR2,TLR4,NF-κB,NLRP3,ASC,Caspase-1 and IL-1β were increased in syphilis group,while the expression of the above indicators was decreased in fixed group(P＜0.05).Compared with fixed group,mRNA ex-pressions of TLR2,TLR4,NF-κB,NLRP3,ASC,Caspase-1 and IL-1β in syphilis group were increased in the syphilis group(P＜0.05).Compared with the control group,IFN-γ and IL-12 levels were decreased in the negative conversion group,syphilis group and fixed group,and IL-13 and IL-10 levels were increased(P＜0.05).Compared with the nega-tive conversion group,IFN-γ and IL-12 levels in the syphilis group and fixed group were decreased,while IL-13 and IL-10 levels were increased(P＜0.05).Compared with the fixed group,the levels of IFN-γ and IL-12 in syphilis group were increased,while the levels of IL-13 and IL-10 were decreased(P＜0.05).The mRNA levels of TLR2,TLR4,NF-κB,NLRP3,ASC,Caspase-1 and IL-1β were positively correlated with the levels of IFN-γ and IL-12 in serum fixed patients with syphilis infection(r=0.698,0.602,0.636,0.702,0.645,0.617,0.583 and 0.669,and 0.693,0.641,0.702,0.623,0.618 and 0.647,respectively,all P＜0.05),and negatively correlated with the levels of IL-13 and IL-10(r=-0.639,-0.705,-0.614,-0.590,-0.598,-0.651 and-0.628,and-0.704,-0.649,-0.606,-0.655,-0.650,-0.641 and-0.539,respectively,all P＜0.05).CONCLUSION The NLRP3 inflammasome pathway and TLRs pathway are abnormally expressed in sero-fast patients with syphilis infection,positively correlated with the expression of Th1 cytokines IFN-γ and IL-12,and negatively correlated with the expression of Th2 cytokines IL-13 and IL-10.

OBJECTIVE To investigate the expression levels of nucleotide-binding oligomerization domain-like recep-tor pyrin domain-associated protein 3(NLRP3)inflammasome pathway and toll-like receptors(TLRs)pathway in serofast patients with syphilis infection,and their correlation with helper T lymphocytes(Th)1/Th2.METHODS A total of 549 patients with syphilis admitted to Tianjin Chest Hospital and Tianjin Medical University General Hospital from Jan.2021 to Dec.2022 were selected.Among them,62 patients who had not received anti-syphilis treatment were assigned to the syphilis group,238 patients whose serum remained positive after anti-syphilis treatment were selected as the fixed group,249 patients whose serum turned negative after anti-syphilis treatment were selected as the negative conversion group,and another 565 healthy people who underwent medical examina-tion in the hospital during the same period were selected as the control group.TLR2,TLR4,nuclear factor(NF)-κB,NLRP3,apoptosis-associated speck-like protein(ASC),cysteinyl aspartate proteolytic enzyme 1(Caspase-1),in-terleukin(IL)-1β mRNA and interferon-γ(IFN-γ),IL-12,IL-13 and IL-10 levels were compared between each group.Pearson's analysis was used to analyze the correlation between the mRNA expression of TLR2,TLR4,NF-κB,NLRP3,ASC,Caspase-1,and IL-1β and the levels of IFN-γ,IL-12,IL-13,and IL-10 in serologically fixed patients with syphilis.RESULTS Compared with the control group,the mRNA expression of TLR2,TLR4,NF-κB,NLRP3,ASC,Caspase-1 and IL-1β was elevated in the negative conversion and the syphilis groups,and the expres-sion of the above indicators was decreased in the fixed group(P＜0.05).Compared with negative conversion group,mR-NA expressions of TLR2,TLR4,NF-κB,NLRP3,ASC,Caspase-1 and IL-1β were increased in syphilis group,while the expression of the above indicators was decreased in fixed group(P＜0.05).Compared with fixed group,mRNA ex-pressions of TLR2,TLR4,NF-κB,NLRP3,ASC,Caspase-1 and IL-1β in syphilis group were increased in the syphilis group(P＜0.05).Compared with the control group,IFN-γ and IL-12 levels were decreased in the negative conversion group,syphilis group and fixed group,and IL-13 and IL-10 levels were increased(P＜0.05).Compared with the nega-tive conversion group,IFN-γ and IL-12 levels in the syphilis group and fixed group were decreased,while IL-13 and IL-10 levels were increased(P＜0.05).Compared with the fixed group,the levels of IFN-γ and IL-12 in syphilis group were increased,while the levels of IL-13 and IL-10 were decreased(P＜0.05).The mRNA levels of TLR2,TLR4,NF-κB,NLRP3,ASC,Caspase-1 and IL-1β were positively correlated with the levels of IFN-γ and IL-12 in serum fixed patients with syphilis infection(r=0.698,0.602,0.636,0.702,0.645,0.617,0.583 and 0.669,and 0.693,0.641,0.702,0.623,0.618 and 0.647,respectively,all P＜0.05),and negatively correlated with the levels of IL-13 and IL-10(r=-0.639,-0.705,-0.614,-0.590,-0.598,-0.651 and-0.628,and-0.704,-0.649,-0.606,-0.655,-0.650,-0.641 and-0.539,respectively,all P＜0.05).CONCLUSION The NLRP3 inflammasome pathway and TLRs pathway are abnormally expressed in sero-fast patients with syphilis infection,positively correlated with the expression of Th1 cytokines IFN-γ and IL-12,and negatively correlated with the expression of Th2 cytokines IL-13 and IL-10.

This study aimed to identify,characterize,and functionally analyze the CYP450 gene fam-ily within Hyalomma asiaticum unfed nymphs(HasiUN)under regular physiological conditions,laying the groundwork for further study into the biosynthesis and metabolism of exogenous sub-stances associated with the CYP450 gene in HasiUN.Through our methods,we identified and characterized HasiUNCYP450 genes.We then annotated them using the gene ontology(GO)and kyoto encyclopedia of genes and genomes(KEGG).This was achieved with the help of bioinforma-tics,based on the transcriptome of 2-week-old HasiUN.Further on,we conducted a preliminary a-nalysis of tissue expression,distribution,and potential function with qRT-PCR and network inter-action predictions.Our analyses showed that 87 HasiUN CYP450 genes,ranging in size from 1089 to 1692 bp,encoded predominantly basic,hydrophilic,and unstable protein sequences.These se-quences had nearly identical numbers of transmembrane regions,both with and without a signal peptide.Phylogenetic and domain analyses indicated that HasiUN CYP450,having a complete CYP450 domain,were distributed among various clans,with most members found in clan 3.CYP20 clans,present in 13 species from 2 families and 7 genera,required either one-host or three-hosts to undergo their full life cycle.MEME analysis results hint at different clans having variations in the amount and combination of motifs.Expression level analysis showed that HasiUN CYP450 was expressed at different levels,with the highest distribution of normally expressed genes in clan 3 and low expression of all mito clan genes.Functional annotation revealed HasiUN CYP450 key as-sociation with biological processes,metabolism,and organismal systems according to the GO and KEGG annotations.Distribution results showed that HasiUN CYP20a1 was expressed in specific tissues,including salivary glands and ovaries.Functional analysis points to CYP20a1's role in UV-induced toxic metabolism,endocrine disruptor metabolism,and hazardous substance metabolism in the nervous system.Our research presents the first analysis of HasiUN CYP450 gene's biological characteristics and expression pattern under normal physiological conditions.Additionally,investi-gations into the distribution and function of a new category,HasiUN CYP20a1,were embarked upon.These findings establish a theoretical foundation for continued functional study of HasiUN CYP45 gene.

Objective:To explore risk factors for delayed onset of lactogenesis in prenatally overweight women and construct risk prediction models based on machine learning for early identification of high-risk individuals.Methods:Convenience sampling was adopted to select 338 prenatally overweight women who delivered in the Obstetrics Departments of four ClassⅢ Grade A hospitals in Hengyang City from October 2023 to June 2024 for the study. Delivery women were randomly divided into training set and test set in the ratio of 7∶3. The survey was conducted with the General Information Questionnaire, Breastfeeding Self-efficacy Scale Short Form, Edinburgh Postnatal Depression Scale, LATCH Scale and Pittsburgh Sleep Quality Index. One-way analysis and LASSO regression were used to screen predictors using delayed onset of lactogenesis as the outcome variable. Risk prediction models were constructed based on three machine learning algorithms of Logistic regression, support vector, and random forest, respectively. The models were tuned by ten-fold cross-validation to filter out the best models.Results:Delayed onset of lactogenesis occurred in 140 of 338 prenatally overweight women, an incidence of 41.4%. Among the three predictive model performances, the random forest model had the highest area under the receiver operating characteristic curve, accuracy, precision, recall, and F1 value. The importance of each predictor was ranked according to the fandom forest algorithm, and in descending order of importance, they were breastfeeding 1 h after the birth of the newborn, number of previous deliveries, age, feeding mode 3 d postpartum, pregnancy complications, mode of delivery, number of breastfeeding 24 h postpartum, and monthly household income.Conclusions:Risk prediction models for delayed onset of lactogenesis in prenatally overweight women are constructed based on three machine learning algorithms, aiming to help provide a scientific basis for clinical healthcare professionals to take relevant decisions.

ObjectiveThis study aims to investigate the molecular mechanism by which Guizhi Fulingwan （GFW） inhibits ferroptosis in endometriosis （EMT） through the regulation of the hypoxia inducible factor-1α/heme oxygenase 1 （HIF-1α/HO-1） signaling pathway. MethodsMachine learning was employed to identify ferroptosis-related biomarkers associated with EMT. Network pharmacology was utilized to identify the active components of GFW and its potential therapeutic targets against EMT， including core targets. Functional enrichment analysis was conducted to explore the biological processes， molecular functions， cellular components， and signaling pathways associated with the potential targets. An EMT rat model was established via autologous transplantation. Thirty female Sprague-Dawley （SD） rats were randomly divided into five groups： sham-operated， model， positive control （dienogest at 0.2 mg·kg^-1）， low-dose GFW （2.5 g·kg^-1）， and high-dose GFW （5 g·kg^-1）. After modeling， the rats received their respective treatment by oral gavage for 28 consecutive days， while the sham and model groups received equal volumes of distilled water. Serum and ectopic endometrial tissues were collected. Hematoxylin and eosin （HE） staining was employed to evaluate morphological alterations in ectopic lesions. Quantitative real-time polymerase chain reaction （Real-time PCR） and Western blot were conducted to assess mRNA and protein expression of HIF-1α， HO-1， glutathione peroxidase 4 （GPX4）， spermidine/spermine N1-acetyltransferase （SAT1）， and prostaglandin-endoperoxide synthase 2 （PTGS2）. Tissue levels of malondialdehyde （MDA）， glutathione （GSH）， and ferrous iron （Fe²⁺） were quantified using commercial assay kits. Serum levels of interleukin-6 （IL-6） and transforming growth factor-β₁ （TGF-β₁） were measured via enzyme-linked immunosorbent assay （ELISA）. ResultsFive ferroptosis-related biomarkers in EMT were identified： ALOX12， CHAC1， SAT1， AST1， and HO-1. Network pharmacology analysis revealed 42 active components of GFW and 192 potential therapeutic target genes related to EMT treatment， with FOS， JUN， HO-1 identified as core targets. Functional enrichment analysis indicated that the potential targets were primarily involved in oxidative stress response and reactive oxygen species metabolism and were enriched in the HIF-1 signaling pathway. Compared to the sham-operated group， the model group exhibited significant increases in both mRNA and protein expression of HIF-1α， HO-1， and PTGS2， as well as elevated tissue levels of Fe²⁺ and MDA. Conversely， GSH levels and the expression of GPX4 and SAT1 were markedly reduced， and serum levels of IL-6 and TGF-β₁ levels were significantly higher （P<0.01）. Compared with the model group， all GFW-treated groups showed significant downregulation of HIF-1α and HO-1， reduced Fe²⁺ levels， and downregulated expression of MDA， PTGS2， IL-6， and TGF-β₁. Meanwhile， GSH， GPX4， and SAT1 expression levels were significantly increased （P<0.05， P<0.01）， effectively ameliorating iron overload and oxidative stress， thereby demonstrating therapeutic efficacy in EMT， with the high-dose GFW demonstrating the most pronounced therapeutic effects. ConclusionGFW exerts therapeutic effects on endometriosis by regulating the HIF-1α/HO-1 signaling pathway to rectify iron metabolism disorders and attenuate free iron-induced oxidative damage. It upregulates the antioxidative defense system to inhibit lipid peroxidation cascades and modulates inflammatory cytokine networks. These effects collectively disrupt the pathological interaction between ferroptosis and chronic inflammation， providing a novel theoretical foundation for the clinical application of GFW in EMT treatment.

Objective:To explore risk factors for delayed onset of lactogenesis in prenatally overweight women and construct risk prediction models based on machine learning for early identification of high-risk individuals.Methods:Convenience sampling was adopted to select 338 prenatally overweight women who delivered in the Obstetrics Departments of four ClassⅢ Grade A hospitals in Hengyang City from October 2023 to June 2024 for the study. Delivery women were randomly divided into training set and test set in the ratio of 7∶3. The survey was conducted with the General Information Questionnaire, Breastfeeding Self-efficacy Scale Short Form, Edinburgh Postnatal Depression Scale, LATCH Scale and Pittsburgh Sleep Quality Index. One-way analysis and LASSO regression were used to screen predictors using delayed onset of lactogenesis as the outcome variable. Risk prediction models were constructed based on three machine learning algorithms of Logistic regression, support vector, and random forest, respectively. The models were tuned by ten-fold cross-validation to filter out the best models.Results:Delayed onset of lactogenesis occurred in 140 of 338 prenatally overweight women, an incidence of 41.4%. Among the three predictive model performances, the random forest model had the highest area under the receiver operating characteristic curve, accuracy, precision, recall, and F1 value. The importance of each predictor was ranked according to the fandom forest algorithm, and in descending order of importance, they were breastfeeding 1 h after the birth of the newborn, number of previous deliveries, age, feeding mode 3 d postpartum, pregnancy complications, mode of delivery, number of breastfeeding 24 h postpartum, and monthly household income.Conclusions:Risk prediction models for delayed onset of lactogenesis in prenatally overweight women are constructed based on three machine learning algorithms, aiming to help provide a scientific basis for clinical healthcare professionals to take relevant decisions.

Objective:To investigate the technical application pathways of artificial intelligence(AI)in laboratory information management systems(LIMS)and its role in promoting laboratory management efficiency and intelli-gence.Methods:Through the integration of traditional AI technologies(e.g.,machine learning,computer vision)with large language models,this study demonstrated the application of various AI technologies in scenarios such as intelligent Q&A for local knowledge bases,comprehensive review of inspection processes,intelligent data visualization,and image recognition.Results:Through the implementation of AI applications in laboratory settings,AI significantly enhanced management efficiency:the intelligent Q&A system achieved over 90％accuracy,auto-mated inspection processes reduced manual workload by 40％,and image recognition precision reached 89％-100％.Conclusion:AI provides efficient and precise solutions for laboratory management via multimodal integration and process optimization.Future efforts should focus on strengthening data security and model interpret-ability to promote comprehensive intelligent development.

Objective To investigate the effects of Echinococcus multilocularis infection on levels of memory T (Tm) cells and their subsets in lymph nodes of mice at different stages of infection, so as to provide new insights into immunotherapy for alveolarechinococcosis. MethodsTwenty-four C57BL/6J mice aged 6 to 9 weeks were randomly divided into the infection group and the control group, of 12 mice in each group. Mice in the infection group were administered with 3 000 E. multilocularis protoscoleces via portal venous injection, while animals in the control group were administered with an equal volume of physiological saline. Three mice from each group were sacrificed 4, 12 weeks and 24 weeks post-infection, and lymph nodes were sampled and stained with hematoxylin and eosin (HE) to investigate the histopathological changes of mouse lymph nodes in the infection group. The expression and localization of T lymphocyte surface markers CD3, CD4, and CD8 were observed in mouse lymph nodes using immunohistochemical staining. In addition, lymphocyte suspensions were prepared from mouse lymph nodes in both groups at different time points post-infection, and the levels of Tm cell subsets and their secreted cytokines were detected using flow cytometry. Results HE staining showed diffuse structural alterations in the subcapsular cortical and paracortical regions of mouse lymph nodes in the infection group 4 weeks post-infection with E. multilocularis. Immunohistochemical staining detected CD3, CD4 and CD8 expression in mouse lymph nodes in both groups. Flow cytometry revealed higher proportions of CD4⁺ Tm cells [(55.3 ± 4.8)% vs. (38.8 ± 6.1)%; t = -4.259, P < 0.05] and CD4⁺ tissue-resident Tm (Trm) cells [(57.7 ± 3.7)% vs. (34.1 ± 11.2)%; t = -3.990, P < 0.05] in mouse lymph nodes in the infection group than in the control group 4 weeks post-infection, and higher proportions of CD4⁺ Tm cells [(34.6 ± 3.2)% vs. (23.3 ± 7.5)%; t = -2.764, P < 0.05] and CD4⁺ Trm cells [(44.0 ± 1.9)% vs. (31.2 ± 1.5)%; t = -4.039, P < 0.05] in mouse lymph nodes in the infection group than in the control group 24 weeks post-infection. The proportions of CD8⁺ Tm cells were higher in the infection group than in the control group 4 weeks [(56.8 ± 2.7)% vs. (43.9 ± 5.2)%; t = -4.416, P < 0.01] and 12 weeks post-infection [(25.4 ± 2.7)% vs. (12.0 ± 2.6)%; t = -2.552, P < 0.05], while the proportions of tumor necrosis factor (TNF)-α⁺ CD4⁺ T cells [(15.7 ± 5.0)% vs. (49.4 ± 6.4)%; t = 7.150, P < 0.01], TNF-α⁺CD8⁺ T cells [(20.7 ± 5.5)% vs. (57.5 ± 8.4)%; t = -6.694, P < 0.01], and TNF-α⁺ CD8⁺ Tm cells [7.0% (1.0%) vs. 31.0% (11.0%); Z = -2.236, P < 0.05] were lower in the infection group than in the control group 24 weeks post-infection. Conclusions Tm cells levels are consistently increased in lymph nodes of mice at different stages of E. multilocularis infection, with Trm cells as the predominantly elevated subset. The impaired capacity of CD8⁺ Tm cells to secrete the effector molecule TNF-α in mouse lymph nodes at the late-stage infection may facilitate chronic parasitism of E. multilocularis.

Combined with elastic network model (ENM), the perturbation response scanning (PRS) has emerged as a robust technique for pinpointing allosteric interactions within proteins. Here, we proposed the PRS analysis of drug-target networks (DTNs), which could provide a promising avenue in network medicine. We demonstrated the utility of the method by introducing a deep learning and network perturbation-based framework, for drug repurposing of multiple sclerosis (MS). First, the MS comorbidity network was constructed by performing a random walk with restart algorithm based on shared genes between MS and other diseases as seed nodes. Then, based on topological analysis and functional annotation, the neurotransmission module was identified as the "therapeutic module" of MS. Further, perturbation scores of drugs on the module were calculated by constructing the DTN and introducing the PRS analysis, giving a list of repurposable drugs for MS. Mechanism of action analysis both at pathway and structural levels screened dihydroergocristine as a candidate drug of MS by targeting a serotonin receptor of serotonin 2B receptor (HTR2B). Finally, we established a cuprizone-induced chronic mouse model to evaluate the alteration of HTR2B in mouse brain regions and observed that HTR2B was significantly reduced in the cuprizone-induced mouse cortex. These findings proved that the network perturbation modeling is a promising avenue for drug repurposing of MS. As a useful systematic method, our approach can also be used to discover the new molecular mechanism and provide effective candidate drugs for other complex diseases.

The focus of green analytical chemistry (GAC) is to minimize the negative impacts of analytical procedures on human safety, human health, and the environment. Several factors, such as the reagents used, sample collection, sample processing, instruments, energy consumed, and the quantities of hazardous materials and waste generated during analytical procedures, need to be considered in the evaluation of the greenness of analytical assays. In this study, we propose a greenness evaluation metric for analytical methods (GEMAM). The new greenness metric is simple, flexible, and comprehensive. The evaluation criteria are based on both the 12 principles of GAC (SIGNIFICANCE) and the 10 factors of sample preparation, and the results are presented on a 0-10 scale. The GEMAM calculation process is easy to perform, and its results are easy to interpret. The output of GEMAM is a pictogram that can provide both qualitative and quantitative information based on color and number.