ObjectiveTo systematically compare the differential regulation of the maternal-fetal interface cell lineages and communication networks in the CBA/J×DBA/2 mouse model of recurrent pregnancy loss （RPL） by the two classic therapeutic methods-tonifying the kidney to stabilize the fetus and invigorating the spleen to stabilize the fetus （Shoutai Wan， Juyuan Jian）-of traditional Chinese medicine （TCM） at the single-cell resolution and clarify their modern scientific connotations. MethodsFemale non-pregnant CBA/J mice were caged with male BALB/c （blank group） and DBA/2 （modeling group） mice separately. Pregnant mice in the modeling group were randomly grouped as follows： high/low-dose Shoutai Wan， high/low-dose Juyuan Jian， model （RPL）， and positive control （dydrogesterone）， with 10 mice in each group. Starting from the day after the detection of the vaginal plug， mice were administrated with drugs or an equal volume of normal saline by gavage for 10 consecutive days. After the intervention， the following indicators were measured. ① Macroscopic evaluation： general conditions， uterine wet weight， embryo loss rate， four coagulation parameters ［prothrombin time （PT）， activated partial thromboplastin time （APTT）， fibrinogen （FIB）， and thrombin time （TT）］， and peripheral blood estradiol （E₂） and progesterone （Pg） levels. The decidua with embryos was stained with hematoxylin-eosin （HE） and evaluated by transmission electron microscopy （TEM）. The expression of B-cell lymphoma-2 （Bcl-2）， vascular endothelial growth factor （VEGF）， angiotensin Ⅱ （AngⅡ）， matrix metalloproteinase-2 （MMP-2）， interleukin-6 （IL-6）， leukemia inhibitory factor （LIF）， CXC chemokine ligand 12 （CXCL12）， and microtubule-associated protein 1 light chain 3 homolog （LC3）Ⅰ/Ⅱ was quantified by Western blot. ② Mechanism analysis at the single-cell level： The decidua with embryos from the blank， model， high-dose Shoutai Wan， and high-dose Juyuan Jian groups （6 mice per group， with 3 single-cell samples per group， totaling 24 mice） were analyzed by the BD Rhapsody™ platform， and the whole-cell atlas was drawn by uniform manifold approximation and projection （UMAP） dimensionality reduction clustering combined with the single-cell mouse cell atlas （scMCA）. The differentially expressed genes （DEGs） and cell interaction networks were analyzed via Gene Ontology （GO）， Kyoto Encyclopedia of Genes and Genomes （KEGG）， and CellChat， and the protein-protein interaction （PPI） map of subtype cells was constructed. The CytoTRACE pseudo-temporal analysis was performed to explore the developmental trajectories of core immune cells （natural killer cells， NK cells） from maternal and fetal sources. Results① Pathological and Western blot results indicated that compared with the blank group， the RPL group showed an increase in the embryo loss rate （P<0.01）， down-regulated expression of Bcl-2， LIF， MMP-2， and Vegf in the decidua with embryos （P<0.05）， up-regulated protein levels of CXCL-12， AngⅡ， and IL-6 （P<0.05）， blocked angiogenesis， apoptosis-inflammation imbalance， and coagulation dysfunction. Both prescriptions dose-dependently reduced the abortion rate and restored the angiogenesis-inflammation balance， and Shoutai pill showed superior performance in restoring the E₂ level to the Pg level （P<0.05）. ② Single-cell transcriptome analysis indicated that compared with the blank group， the RPL group showed differences in multiple key cell populations such as decidual cells， trophoblast cells， endothelial cells， erythroblasts， NK cells， and macrophages at the maternal-fetal interface. Immunity and angiogenesis were the key links in RPL. Compared with the RPL group， high-dose Shoutai Wan reversed the changes of NK cells in the embryonic layer （upregulating the mRNA levels of 17 genes and downregulating the mRNA levels of 29 genes） and macrophages （upregulating the mRNA levels of 117 genes and downregulating the mRNA levels of 53 genes） through the regulation of gene expression. High-dose Shoutai pill regulated the immune cells to affect unfolded proteins， cell adhesion， and programmed cell death， thereby promoting decidualization and angiogenesis and modulating embryo-membrane development. High-dose Juyuan Jian regulated the key subgroups of NK cells （up-regulating the mRNA levels of 9 genes and down-regulating the mRNA levels of 17 genes） and macrophages （up-regulating the mRNA levels of 110 genes and down-regulating the mRNA levels of 81 genes）， which affected decidual inflammation and apoptosis and intervened in glycolysis. ③ The pseudo-temporal analysis and communication network indicated that the communication frequency of the RPL group decreased. High-dose Shoutai Wan restored maternal-fetal tolerance through pathways such as NKG2D， CDH5， GDF， and FASLG. High-dose Juyuan Jian enhanced the IL-6/LIFR/JAK/signal transducer and activator of transcription 3 （STAT3） and desmosome/SEMA6/tumor necrosis factor-like weak inducer of apoptosis （TWEAK） signaling to improve endometrial receptivity. The RPL group showed an increased proportion of toxic dNK7， a decreased proportion of reparative dNK4， and blocked embryo fNK1. High-dose Shoutai Wan down-regulated dNK7 and up-regulated dNK4. High-dose Juyuan Jian inhibited the terminal differentiation of dNK7 and up-regulated LILRB1， thus restoring the balance of cytotoxicity and repair. ConclusionBoth the kidney-tonifying and spleen-invigorating methods are effective in treating RPL. NK and macrophages are the key immune cells in the interaction between the embryo and the membrane. The kidney-tonifying method （Shoutai Wan） has an advantage in regulating the phenotypes of unfolded protein， cell adhesion， and programmed cell death， and shows expression characteristics closer to the physiological state in the regulation of NKG2D and CDH5 signals. The spleen-invigorating method （Juyuan Jian） has an advantage in regulating epithelial-mesenchymal transition （EMT）， angiogenesis， and glycolysis and shows higher communication intensity in the IL-6 and LIFR pathways.

Risk prediction models for postoperative pulmonary complications (PPCs) can assist healthcare professionals in assessing the likelihood of PPCs occurring after surgery, thereby supporting rapid decision-making. This study evaluated the merits, limitations, and challenges of these models, focusing on model types, construction methods, performance, and clinical applications. The findings indicate that current risk prediction models for PPCs following lung cancer surgery demonstrate a certain level of predictive effectiveness. However, there are notable deficiencies in study design, clinical implementation, and reporting transparency. Future research should prioritize large-scale, prospective, multi-center studies that utilize multiomics approaches to ensure robust data for accurate predictions, ultimately facilitating clinical translation, adoption, and promotion.

Objective To investigate the clinical efficacy of modified Wuzi Yanzong Pills combined with sequential therapy for the treatment of patients with premature ovarian failure(POF)of yang deficiency of spleen and kidney type,and to observe their effects on sex hormones and ovarian ultrasound indicators.Methods From September 2022 to September 2023,a total of 62 patients with POF of yang deficiency of spleen and kidney type who attended Huizhou Hospital of Traditional Chinese Medicine were selected as the study objects.The patients were randomly divided into the control group and the observation group by using a table of random numbers,with 31 patients in each group.The control group was treated with estrogen-progestin sequential therapy,and the observation group was treated with modified Wuzi Yanzong Pills on the basis of treatment for the control group.The two groups were treated for three menstrual cycles.Before and after treatment,the changes of serum sex hormones,uterine and ovarian ultrasonography and hemodynamic indicators,traditional Chinese medicine(TCM)syndrome scores,Kupperman score of the two groups were observed.The ovulation rate and therapeutic efficacy of the two groups were compared,and the safety of the therapeutic regimen for the two groups was evaluated.Results(1)Two cases fell off during the study period,and a total of 60 patients were eventually included in the study,of which 30 patients in each group.(2)After three menstrual cycles of treatment,the total effective rate of the observation group was 90.00％(27/30)and that of the control group was 76.67％(23/30),and the total effective rate of the observation group was significantly higher than that of the control group(P＜0.05).(3)After treatment,the serum follicle-stimulating hormone(FSH)and luteinizing hormone(LH)levels of the two groups of patients were decreased compared with those before treatment(P＜0.05),and the serum estradiol(E2)level was increased compared with that before treatment(P＜0.05),the decrease of the serum FSH and LH levels and the increase of the serum E2 level in the observational group were significantly superior to those of the control group(P＜0.01).(4)After treatment,ultrasonographic indicators such as endometrial thickness(EM),ovarian volume averaged(OVA),mean ovarian diameter(MOD)at the maximum transverse section of the ovary,and antral follicle count(AFC)in the two groups were increased compared with those before treatment(P＜0.05),and the increase in the observation group was significantly superior to that in the control group(P＜0.01).(5)After treatment,the peak systolic velocity(PSV)of the two groups was increased compared with that before treatment(P＜0.05),and the ovarian resistance index(RI)and ovarian pulsatility index(PI)were decreased compared with those before treatment(P＜0.05),and the increase of PSV and the decrease of RI and the PI in the observation group were significantly superior to those of the control group(P＜0.01).(6)After treatment,the Kupperman score and TCM syndrome scores of the two groups were decreased compared with those before treatment(P＜0.05),and the decrease in the observation group was significantly superior to that in the control group(P＜0.01).(7)After three menstrual cycles of treatment,the ovulation rate of the observation group was 30.00％(9/30),and that of the control group was 10.00％(3/30).The comparison between the two groups showed that the ovulation rate of the observation group was significantly superior to that of the control group(P＜0.05).(8)The incidence of adverse reactions in the control group was 6.67％(2/30)and that in the observation group was 6.67％(2/30).The comparison between the two groups showed that there being no statistically significant difference(P＞0.05).Conclusion The modified Wuzi Yanzong Pills combined with estrogen-progestin sequential therapy can effectively regulate the sex hormone levels of patients with POF of yang deficiency of spleen and kidney type,promote the follicular development,significantly improve the ovarian microcirculation,increase the ovulation rate,and relieve symptoms and signs,with good safety.

Objective To explore the value of preoperative amide proton transfer(APT)imaging combined with diffusion kurtosis imaging(DKI)in predicting lymphovascular space invasion(LVSI)in cervical cancer.Methods Fifty-one patients with cervical cancer who underwent preoperative MRI examination and had complete postoperative pathology were retrospectively selected.Pelvic MRI scans were performed 1-2 weeks preoperatively,and the corresponding APT values,mean kurtosis(MK)and mean diffusivity(MD)values were obtained respectively,and the occurrence of LVSI was determined based on postoperative pathology results.The predictive effects of APT-and DKI-derived parameters alone or in combination on the LVSI status of cervical cancer were compared.Results Among 51 cases of cervical cancer,36 cases had pathologically confirmed LVSI and 15 cases did not had LVSI.The area under the curve(AUC)of the receiver operating characteristic(ROC)curve for the preoperative APT values,MK and MD values alone and in combination to predict the LVSI status of cervical cancer were 0.820,0.788,0.762,0.894,0.885,and 0.896,respectively.APT values combined with DKI-derived parameters predicted LVSI better than when they were used separately,in which APT values combined with MK and MD values predicted LVSI of cervical cancer with the largest AUC,sensitivity of 91.7％,specificity of 80.0％,and Youden's index of 0.717.Conclusion Preoperative APT imaging and DKI have important value in predicting LVSI of cervical cancer,and the combined application of the two can improve the prediction efficacy,which has certain clinical application value.

Objective:To investigate the efficacy of antiviral therapy and influencing factors of hepatitis B surface antigen(HBsAg) negative conversion for hepatitis B e antigen(HBeAg)-positive chronic hepatitis B(CHB) in children.Methods:The clinical data of 38 children with CHB who received antiviral treatment in Children's Hospital Affiliated to Xi'an Jiaotong University from January 2019 to August 2024 were collected.All patients were treated with interferon alpha monotherapy or combined with nucleoside analogues for 48 weeks.The patients were divided into HBsAg negative group and HBsAg non-negative group according to the therapeutic results at 48 weeks.Multivariate Logistic regression were used to identify influencing factors of HBsAg negative conversion at 48 weeks.The receiver operator characteristic（ROC）curve was used to analyze the predictive value of each factor to HBsAg negative conversion.Results:The alanine aminotransferase normalization rate，hepatitis B virus DNA negative rate，HBeAg negative rate and HBsAg negative rate were 76.3%，94.7%，39.5% and 47.4%，respectively at 48 weeks.There were 18 cases in HBsAg negative group and 20 cases in HBsAg non-negative group.There were statistical significant differences in age and HBsAg decline level at 12 and 24 weeks of antiviral treatment between HBsAg negative group and HBsAg non-negative group（ P<0.05）.Multivariate Logistic regression analysis showed that age and HBsAg decline level at 12 and 24 weeks of antiviral treatment were independent predictors of HBsAg negative conversion at 48 weeks（ OR=0.664，95% CI 0.473-0.932， P=0.018； OR=8.719，95% CI 1.920-39.604， P=0.005； OR=6.182，95% CI 2.083-18.347， P=0.001）.The area under the curve of age and HBsAg decline level at 12 and 24 weeks were 0.737（95% CI 0.576-0.899， P=0.012），0.847（95% CI 0.725-0.969， P<0.001）and 0.939（95% CI 0.811-0.991， P<0.001），respectively.When the age was less than 4.625 years，the sensitivity，specificity，positive predictive value and negative predictive value of HBsAg negative conversion at 48 weeks were 83.3%，65.0%，68.2% and 81.3%，respectively.A decrease in HBsAg level of >1.07 lg IU/mL at 12 weeks of treatment had a sensitivity，specificity，positive predictive value，and negative predictive value of 72.2%，90.0%，86.7%，and 78.3%，respectively，for predicting HBsAg seroclearance at 48 weeks.A reduction in HBsAg of >1.92 lg IU/mL at 24 weeks of treatment showed a sensitivity，specificity，positive predictive value，and negative predictive value of 83.3%，90.0%，88.2%，and 85.7%，respectively，in predicting HBsAg seroclearance at 48 weeks. Conclusion:The children with CHB have a higher rate of HBsAg negative conversion after antiviral therapy at 48 weeks.Age and HBsAg decline level at 12 and 24 weeks of antiviral treatment can serve as early predictors for HBsAg negative conversion in children with CHB.

Objective:To analyze clinical characteristics of Mycoplasma pneumoniae-induced rash and mucositis in children. Methods:Clinical data were collected from children diagnosed with Mycoplasma pneumoniae -induced rash and mucositis in the Department of Dermatology, Capital Center for Children's Health, Captital Medical University from August 1, 2023, to December 1, 2023, and their clinical features were analyzed. Results:A total of 15 children with Mycoplasma pneumoniae-induced rash and mucositis were collected, including 8 males and 7 females, aged 4 to 12 years. They were clinically characterized by obvious mucositis, and skin lesions were relatively mild. Oral mucosal involvement was observed in 13 cases, including lip ulcers (12 cases), oral mucositis (7 cases), tongue ulcers (3 cases), and glossitis (1 case) ; ocular mucosal involvement was reported in 7 cases, including suppurative conjunctivitis (4 cases), and eyelid edema (3 cases) ; 3 children had vulvogenital mucosal involvement, including coronary sulcus ulcers (1 case), urethral meatus ulcers (1 case), labia minora ulcers (1 case), and perianal ulcers (1 case) ; 1 child had nasal mucosal ulcers. Skin lesions occurred on the upper limbs in 5 cases, on the lower limbs in 6 cases, on the face in 2 cases, on the vulva in 2 cases, as well as on the trunk in 1 case. The types of skin lesions included targetoid erythema in 11 cases, vesiculobullous lesions in 6 cases and atypical targetoid erythema in 2 cases. PCR for the detection of Mycoplasma pneumoniae in vesiculobullous fluids showed negative results in the 6 patients with vesiculobullous lesions. The patients received systemic and topical treatment with antibiotic agents and glucocorticoids, as well as supportive therapy. Main pulmonary symptoms as well as skin and mucosal lesions of the 15 children subsided within 3 weeks, and they all had a favorable prognosis, without obvious complications or sequelae. Conclusion:Mycoplasma pneumoniae-related skin and mucosal lesions predominantly present as mucositis, with mild or absent skin involvement. The disorder usually has a mild course, characterized by short disease duration and favorable prognosis.

AIM:To investigate the potential mechanism of licorice on liver injury induced by Se-men Strychni based on network pharmacology,mo-lecular docking combined with animal experiments,providing an effective strategy for prevention and treatment of liver injury induced by Semen Strych-ni.METHODS:Firstly,the active ingredients of Se-men Strychni and licorice were obtained through the ETCM,TCMSP database and analysis platform,CTD database and literature supplementation.Then,the potential toxic ingredients of Semen Strychni were further screened based on the Swis-sADME platform,and the targets corresponding to the active ingredients were predicted through the SwissTargetPrediction platform.By using the Gene-Cards and OMIM databases to collect DILI-related targets,the potential targets for licorice to alleviate liver injury caused by Semen Strychni were ob-tained.By constructing the active ingredient-target network,the core ingredients of licorice in alleviat-ing liver injury caused by Semen Strychni were screened.The key targets obtained were used to construct and analyze the protein-protein interac-tion networks(PPI)through the STRING database and Cytoscape 3.9.0 software.The potential targets were subjected to GO and KEGG pathway enrich-ment analysis with the aid of the DAVID database,and constructed a network of active ingredient-tar-get-pathway.Molecular docking study was ap-proved for the core targets and the active ingredi-ents by using Schrodinger 2023-1 software,and the visualization operation was conducted through Py-mol.Finally,the regulatory effect of licorice on the key pathway of liver injury caused by Semen Strych-ni was validated by establishing a rat model of liver injury induced by Semen Strychni.RESULTS:After screening,6 potential toxic components of Semen Strychni and 104 corresponding targets,89 active components of licorice and 347 corresponding tar-gets,and 3 200 DILI targets were obtained.A total of 23 intersection targets were obtained through Venn analysis.By constructing the active ingredi-ent-target network,it was found that the main core ingredients were 7-methoxy-2-methyl isofla-vone,medicarpin,shinpterocarpin,quercetin,for-mononetin and isoliquiritigenin.The PPI network indicated that the core targets were protein kinase B1(AKT1),epidermal growth factor receptor(EG-FR),human epidermal growth factor receptor 2(ERBB2),glycogen synthase kinase 3 beta(GSK3B),kinase insert domain receptor(KDR)and Janus ki-nase 2(JAK2).A total of 39 relevant pathways were enriched in KEGG(P＜0.01),among which the phos-phatidylinositol 3-kinase(PI3K)-protein kinase B(AKT)(PI3K-AKT)signaling pathway,which has been confirmed and ranks first in enrichment,and was closely related to liver injury.Molecular docking re-sults showed that the core components have good binding ability with the core targets.In vivo animal experiments demonstrated that,compared to the model group,licorice significantly reduced the liver index(P＜0.01),serum levels of alkaline phospha-tase(ALP),alanine aminotransferase(ALT),aspar-tate aminotransferase(AST),indirect bilirubin(IBIL),and total bilirubin(TBIL)in rats with liver in-jury,while increasing total protein(TP)levels(P＜0.05 or P＜0.01).Additionally,licorice alleviated con-gestion in the central veins and hepatic sinusoids,improved the alignment of hepatocytes,and re-duced inflammatory cell infiltration.Furthermore,licorice significantly decreased the levels of malo-ndialdehyde(MDA),tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),and interleukin-6(IL-6)in the liver tissue of injured rats,while elevating the levels of superoxide dismutase(SOD)and glutathi-one peroxidase(GSH-Px)(P＜0.01).It also markedly downregulated the phosphorylation levels of PI3K and AKT(P＜0.01).CONCLUSION:Licorice has multi-component and multi-target properties in the treat-ment of liver injury induced by Semen Strychni,which may play a hepatoprotective role by inhibit-ing the activation of PI3K-AKT signaling pathway.

Background and purpose:High-throughput detection of plasma cell-free DNA(cfDNA)is widely used for multi-cancer targeted therapy drug screening,and this study investigated the relationship between the type and number of plasma cfDNA class Ⅰ and Ⅱ targeted therapy-related gene variants and cancer survival in patients with non-small cell lung cancer(NSCLC).Methods:The sequencing results and clinical data of NSCLC patients who underwent tumor plasma cfDNA high-throughput sequencing projects in Sun Yat-sen University Cancer Center from 2021 to 2023 were collected.The survival follow-up of enrolled patients was carried out from the day of plasma collection on June 1,2021 to May 27,2024,and GraphPad Prism 8.0 and SPSS Statistics 25.0 were used.Univariate and multivariate statistical analyses were conducted on the types and numbers of class Ⅰ and class Ⅱ targeted therapy-related genes in the survival and clinical data of patients and sequencing results(Ethical approval:B2024-359-01).Results:A total of 313 patients included in this study with NSCLC were categorized into stage Ⅰ 25 patients(7.98%),stageⅡ 20 patients(6.39%),stage Ⅲ 38patients(12.14%),and stage Ⅳ 230 patients(73.48%).Pathological diagnosis results showed that adenocarcinoma accounted for 90.10%,squamous cell carcinoma accounted for 5.11%,large cell carcinoma accounted for 2.87%and other classifications accounted for 1.92%.The number and the percentage of class Ⅰ and class Ⅱ targeted therapy drug-related genes in the plasma cfDNA NSCLC patients were 0(25.24%),1(17.57%),2(19.17%),3(14.38%),4(8.31%),and 5 or more(15.34%).The results of statistical analysis showed that 3 genes with the highest mutation frequencies were EGFR,TP53 and ERBB2,and the mutation frequency of EGFR gene was 36.04%.The mutation frequency of TP53 gene was 30.63%.The mutation frequency of ERBB2 gene was 4.95%.The survival time of patients is related to not only the expression of hotspot targeted genes,but also the number of class Ⅰ and Ⅱ target-related gene variants detected by plasma cfDNA high-throughput sequencing.The survival time of the patients with no targeted therapy-related locus variants after treatment was longer compares with targeted therapy-related locus variants,which can reduce the risk of death by 63.2%.However,patients with a single gene locus variant had longer survival time and lower risk of death than those with multiple driver locus variants,and the measured class Ⅰ and Ⅱ targeted therapy drugs were within 3 genes.Overall,the smaller the number of genes,the longer the survival.Conclusions:The number of class Ⅰ and class Ⅱtargeted therapy-related gene variants in plasma cfDNA high-throughput sequencing also has an effect on the survival of patients after treatment.Plasma cfDNA level detected by high-throughput sequencing could be a prognostic factor for the NSCLC patients.

Objective:To investigate the hepatoprotective effect of N-acetylcysteine(NAC)on rats with liver injury induced by cisplatin and its effect on intestinal flora and the expression of interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),and nuclear factor-kappa B(NF-κB).Methods:Male Sprague-Dawley rats were randomly divided into control group(CG),cisplatin group(CP),and NAC group.The rats in the NAC group were given NAC 15 mg/kg by gavage for 8 consecutive days.At half an hour after intragastric administration on the fifth day,all rats except those in the NC group were given intraperitoneal injection of 8 mg/kg cisplatin to induce acute liver injury.An automatic biochemical analyzer was used to measure the content of aspartate aminotransferase(AST),alanine aminotransferase(ALT),alkaline phosphatase(ALP),and total bilirubin(TBIL);liver index was calculated for the rats;Western blot was used to measure the relative expression levels of NF-κB,IL-6,and TNF-α in liver tissue;the 16S rDNA technique was used to measure and analyze the amplification information of the V3-V4 regions of each sample.Results:Compared with the NC group,the CP group had significant increases in the content of AST,ALT,ALP,and TBIL,while NAC reversed the abnormal liver function caused by cisplatin.Compared with the NC group,the CP group had a sig-nificant increase in liver index(P=0.000),while the NAC group had a significant reduction in liver index compared with the CP group(P=0.007).Compared with the NC group,the CP group had signifi-cant increases in the expression levels of IL-6,TNF-α,and NF-κB,while the NAC group showed reductions in the expression of these genes,with significant differences in the expression of IL-6 and TNF-α(P=0.006 and 0.000).Compared with the NC group,the CP group had a significant increase in the α-diversity index of intesti-nal flora,while compared with the CP group,the NAC group tended to have a reduction in the α-diversity index of intestinal flora.Com-pared with the CP group at the phylum level,the NAC group had an increase in the abundance of Actinobacteria and a reduction in the abundance of Firmicutes.Compared with the CP group at the genus level,the NAC group had a reduction in the abundance of Rumino-coccaceae and increases in the abundance of Bifidobacterium and Allobaculum.Conclusion:NAC can alleviate acute liver injury caused by cisplatin,possibly by downregulating the expression of IL-6,TNF-α,and NF-κB and regulating the abundance and diver-sity of intestinal flora.

OBJECTIVES: Exposure to rare earth elements (REEs) has been linked to various systemic diseases, but their impact on the offspring blood-brain barrier (BBB) via the gut-brain axis remains unclear. This study aims to investigate the effects of maternal exposure to neodymium oxide (Nd₂O₃) on the BBB integrity of offspring rats, and to evaluate the potential protective role of bifidobacterium tetrad viable tablets (BTVT) against Nd₂O₃-induced intestinal and BBB damage. METHODS: Healthy adult SD rats were mated at a 1:1 male-to-female ratio, with the day of vaginal plug detection marked as gestational day 0. A total of 60 pregnant rats were randomly assigned to the following groups: Control, 50 mg/(kg·d) Nd₂O₃, 100 mg/(kg·d) Nd₂O₃, 200 mg/(kg·d) Nd₂O₃, and 200 mg/(kg·d) Nd₂O₃ + BTVT group. Treatments were administered by daily oral gavage throughout pregnancy and lactation. On postnatal day 21 (weaning), offspring feces, brain, and colon tissues were collected. Hematoxylin and eosin (HE) staining was used to assess structural changes in brain and intestinal tissues. Short-chain fatty acids (SCFAs) in feces were quantified by gas chromatography-mass spectrometry (GC-MS). Evans Blue (EB) dye extravasation assessed BBB permeability. Gene and protein expression levels of tight junction proteins occludin and zonula occludens-1 (ZO-1) were measured by reverse transcription PCR (RT-PCR) and Western blotting (WB), respectively. Neodymium levels in brain tissue were determined via inductively coupled plasma mass spectrometry (ICP-MS). RESULTS: HE staining revealed that maternal Nd₂O₃ exposure caused mucosal edema, increased submucosal spacing, and lymphocyte infiltration in offspring colon, as well as neuronal degeneration and vacuolization in brain tissue. BTVT intervention alleviated these changes. GC-MS analysis showed that levels of acetic acid, propionic acid, butyric acid, and isobutyric acid significantly decreased, while valeric acid and isovaleric acid increased in offspring of Nd₂O₃-exposed mothers (P<0.05). BTVT significantly restored levels of acetic, propionic, and isobutyric acids and reduced valeric acid content (P<0.05). EB permeability was significantly elevated in Nd₂O₃-exposed offspring brains (P<0.05), but reduced with BTVT treatment (P<0.05). RT-PCR and WB showed downregulation of occludin and ZO-1 expression following Nd₂O₃ exposure (P<0.05), which was reversed by BTVT (P<0.05). ICP-MS results indicated significantly increased brain neodymium levels in offspring from all Nd₂O₃-exposed groups (P<0.05), while BTVT significantly reduced neodymium accumulation compared to the 200 mg/(kg·d) Nd₂O₃ group (P<0.05). CONCLUSIONS Maternal exposure to Nd₂O₃ during pregnancy and lactation disrupts intestinal health and BBB integrity in offspring, elevates brain neodymium accumulation, and induces neuronal degeneration. BTVT effectively mitigates Nd₂O₃-induced intestinal and BBB damage in offspring, potentially through modulation of the gut-brain axis.
Animals ; Female ; Blood-Brain Barrier/pathology* ; Pregnancy ; Rats, Sprague-Dawley ; Rats ; Male ; Neodymium/toxicity* ; Prenatal Exposure Delayed Effects/prevention & control* ; Lactation ; Maternal Exposure/adverse effects* ; Brain