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.

ObjectiveTo investigate whether Bushen Huoxue prescription improves embryo implantation through regulating exosomal miRNA to enhance maternal-fetal interface communication based on Bushen Huoxue therapy. MethodsIn the animal experiment， all the rats （except for the blank group） were administered hydroxyurea （450 mg·kg^-1） via gavage for 10 d， as well as epinephrine （0.3 mg·kg^-1） and mifepristone （5.5 mg·kg^-1） via subcutaneous injection for 7 d to establish an implantation disorder model of kidney deficiency and blood stasis type. The Bushen Huoxue prescription （BSHX） groups were administered the prescription at different doses （7.30 g·kg^-1 for the high-dose group， 3.65 g·kg^-1 for the medium-dose group， and 1.83 g·kg^-1 for the low-dose group） via gavage. The dydrogesterone group was administered the corresponding medicine （2.63 mg·kg^-1） via gavage. After intervention for 10 days， uterine histopathological changes were observed via hematoxylin-eosin （HE） staining. Mucin （MUC1）， forkhead box protein O1 （FoxO1）， and homeobox A10 （HoxA10） expression levels were detected by Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and Western blot. Cell experiment selected primary endometrial epithelial cells （EEC） and trophoblast cells （TC） as research subjects. Exosome-free medicated serum was prepared by ultracentrifugation and cultured in complete medium. Exosomes were isolated from cell supernatants by ultracentrifugation for cross-co-culture. After 48 h， migration and invasion abilities were assessed by scratch and Transwell assays. Sequencing was then performed on EEC-origin exosomal miRNA. ResultsThe model rats exhibited thin endometrium， along with reduced blood vessels， glandules， and pinopode numbers. BSHX improved endometrial morphology and increased pinopode numbers. MUC1， FoxO1， and HoxA10 expressions were downregulated in the model rats， while these parameters were upregulated after BSHX medium- and high-dose intervention. In the cell experiment， after exosome-free medicated serum intervention for 24 h， migration and invasion abilities were enhanced in the BSHX groups （P<0.01）. In EEC-origin exosomal miRNA sequencing， gene ontology （GO） and Kyoto Encyclopedia of Genes and Genomes （KEGG） analyses revealed enrichment in biological processes （gastrulation， neuronal differentiation， alongside cell development and regeneration）， involving the mitogen-activated protein kinase （MAPK）， FoxO1， Wnt， mammalian target of rapamycin （mTOR）， and tumor necrosis factor （TNF） signaling pathways. ConclusionBSHX promotes embryo implantation by improving endometrial receptivity via regulating exosomal miRNA. These findings provide potential targets for exosomal miRNA-based assisted reproductive strategies and a novel theoretical basis for infertility treatment by traditional Chinese medicine.

Objective To investigate the protective effects and mechanism of Shuilian Fengkui Formula(SFF)of Yao medicine on sequelae pelvic inflammatory disease(SPID)in rats.Methods The SPID rat model was established by mechanical injury coupled with bacterial infection,and then the rats were randomly divided into:the SPID group,the positive drug(Fukeqianjin tablets)group,SFF groups at low,medium and high doses,and the sham-operation group,with 8 rats in each group;SPID group,SFF groups,TLR4 agonist CRX-527(CRX-527)group,SFF plus CRX-527 group,with 8 rats in each group.Corresponding drug intervention was given for 21 consecutive days.The indexes of uterus,spleen,thymus and other organs were calculated.The histopathological changes in rat uterus were observed by HE staining.The levels of interleukin-1β(IL-1β),interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),immunoglobulin G(IgG),immunoglobulin M(IgM)in serum were detected by ELISA;The protein expression levels of Toll receptor 4(TLR4),myeloid differentiation factor(MyD88),nuclear factor kappa B p65(NF-κB p65),and phosphorylated nuclear factor kappa B p65(p-NF-κB p65)in rat uterine tissues were detected by Western blot.Results Compared with the sham operation group,the uterine index of SPID group significantly increased(P<0.01),the spleen index and thymus index significantly decreased(P<0.01),the pathological damage of uterine tissue was serious,the contents of IL-1β,IL-6,TNF-α in serum significantly increased(P<0.01),the contents of IgG and IgM significantly decreased(P<0.01),the TLR4,MyD88 protein and the ratio of p-NF-κB p65/NF-κB p65 protein in uterine tissue significantly increased(P<0.01).Compared with the SPID group,the uterine index of the positive drug group and the SFF groups significantly decreased(P<0.05),the spleen index and thymus index significantly increased(P<0.05),the pathological damage of uterine tissue significantly improved,the contents of IL-1β,IL-6 and TNF-α in serum were significantly lowered(P<0.05),the contents of IgG and IgM significantly increased(P<0.05),and the TLR4,MyD88 protein and the ratio of p-NF-κB p65/NF-κB p65 protein in uterine tissue significantly downregulated(P<0.01).However,CRX-527 significantly reversed the effects of SFF on SPID in rats.Conclusion SFF could improve SPID in rats by impeding the inflammatory response,and its mechanism might be attributed to inhibition of TLR4/MyD88/NF-κB signaling pathway.

Objective:To explore the application value of right-opening single flap valvuloplasty based on tubular stomach in gastrointestinal reconstruction after laparoscopic proximal gastrectomy.Method:Use a linear cutting stapler to make a parallel curve from the angle of the stomach to the junction of the gastric fundus to remove the lesser curvature of the stomach, and detach the gastric body about 5 cm away from the tumor to create a tubular stomach. Use a marker pen to draw a C-shaped seromuscular flap area with a width of 2.5 cm and a height of 3.5 cm 1.5 cm below the residual stomach closure nail, and create a free muscle flap in the gap between the plasma muscle layer and the submucosal layer. Make a transverse incision of 3 cm at the lower edge of the mucosal bed, and intermittently suture the entire lower edge of the gastric wall with 3 stitches. Under laparoscopy, use 4-0 barbed wire to suture the 1 cm wide muscular layer at the top of the tubular stomach and the posterior wall of the esophagus about 5 cm away from the esophageal stump with 3 stitches. Push the upper end of the tubular stomach into the mediastinum, and then tighten the barbed wire to ensure a tight fit between the stomach and the posterior wall of the esophagus. Use an ultrasonic scalpel to remove the esophageal stump, suture the entire posterior wall of the esophagus with the gastric mucosa, and use barbed wire to suture the anterior wall from left to right. The anastomotic site is completely covered with a free muscle flap, and the barbed line is used to continuously suture the muscle flap along the C-shaped line to the gastric pulp muscle layer at the edge of the mucosal bed, embedding the anastomotic site and completing the reconstruction of the digestive tract.Results:Clinical data of 23 patients (18 from the First Affiliated Hospital of Wenzhou Medical University and 5 from the Quzhou Hospital affiliated with Wenzhou Medical University) who underwent laparoscopic proximal gastrectomy, tubular gastroesophageal anastomosis, and pure manual right flap reconstruction surgery for esophagogastric junction adenocarcinoma and proximal gastric cancer from October 2023 to August 2024. There were 15 males and 8 females, with an age of (65.3±7.7) years, the BMI was (22.9±2.8) kg/m 2. All patients in the group successfully completed the surgery, with a surgery time of (218.5±38.1) minutes, including (73.5±19.2) minutes for anastomosis, intraoperative blood loss of (64.5±15.4) ml, postoperative passage of gas on (3.4±0.5) days, first consumption of liquid food after surgery of (3.9±1.1) days, and postoperative hospital stay of (9.1±0.8) days. One patient developed anastomotic stenosis (grade I) after surgery, presenting with mild swallowing obstruction, which returned to normal after dietary adjustment, and there were no cases of secondary surgery. The median follow-up time for the entire group was 4.0 (0.7-7.0) months, during which there were no deaths or tumor recurrence or metastasis, no complications such as anastomotic stenosis or gastric emptying disorders, and no complaints of acid reflux or heartburn. At one month of postoperative follow-up, the reflux symptom index (RSI) score was (3.1±2.9) points, and at three months, the RSI score was (2.4±1.4) points. Conclusions:The application of right-opening single flap valvuloplasty based on tubular stomach for gastrointestinal reconstruction after laparoscopic proximal gastrectomy is safe,feasible,and has satisfactory short-term efficacy.

Objective:To analyze and validate the reliability and validity of the social skills improvement system-rating scales Chinese version (parent version) (SSIS-RS-C) in middle school students.Method:A total of 1 486 parents of middle school students were recruited according to the cluster sampling method.The social responsiveness scale and strengths and difficulties questionnaire were used as criterion validity tools.A retest was conducted one month later.SPSS 27.0 was used for descriptive statistics, item analysis, internal consistency test, test-retest reliability test and criterion validity test. AMOS 24.0 was used to perform confirmatory factor analysis .Results:Item analysis indicated significant positive correlations between each item and the subscales ( r=0.293-0.782, all P<0.01), with significant differences in scores between high and low groups ( t=10.079-37.038, all P<0.01).Confirmatory factor analysis supported a seven-factor structure for the social skills subscale(communication, cooperation, assertion, responsibility, empathy, engagement and self control) and a five-factor structure for the problem behavior subscale (externalizing, bullying, hyperactivity/inattention, internalizing and autism spectrum) of the SSIS-RS-C.There was a positive correlation between the social skills subscale and prosocial behavior ( r=0.637, P<0.001), and between the problem behavior subscale and social impairments and difficult behaviors ( r=0.765, 0.688, both P<0.001).The Cronbach's α coefficients for the total scale, social skills subscale and problem behavior subscale were 0.934, 0.972 and 0.963, respectively.The test-retest correlation coefficients for the total score and the two subscales were 0.665, 0.871 and 0.598, respectively (all P<0.001). Conclusion:The SSIS-RS-C demonstrated good reliability and validity in the Chinese adolescent population.

Objective To investigate the protective effects and mechanism of Shuilian Fengkui Formula(SFF)of Yao medicine on sequelae pelvic inflammatory disease(SPID)in rats.Methods The SPID rat model was established by mechanical injury coupled with bacterial infection,and then the rats were randomly divided into:the SPID group,the positive drug(Fukeqianjin tablets)group,SFF groups at low,medium and high doses,and the sham-operation group,with 8 rats in each group;SPID group,SFF groups,TLR4 agonist CRX-527(CRX-527)group,SFF plus CRX-527 group,with 8 rats in each group.Corresponding drug intervention was given for 21 consecutive days.The indexes of uterus,spleen,thymus and other organs were calculated.The histopathological changes in rat uterus were observed by HE staining.The levels of interleukin-1β(IL-1β),interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),immunoglobulin G(IgG),immunoglobulin M(IgM)in serum were detected by ELISA;The protein expression levels of Toll receptor 4(TLR4),myeloid differentiation factor(MyD88),nuclear factor kappa B p65(NF-κB p65),and phosphorylated nuclear factor kappa B p65(p-NF-κB p65)in rat uterine tissues were detected by Western blot.Results Compared with the sham operation group,the uterine index of SPID group significantly increased(P<0.01),the spleen index and thymus index significantly decreased(P<0.01),the pathological damage of uterine tissue was serious,the contents of IL-1β,IL-6,TNF-α in serum significantly increased(P<0.01),the contents of IgG and IgM significantly decreased(P<0.01),the TLR4,MyD88 protein and the ratio of p-NF-κB p65/NF-κB p65 protein in uterine tissue significantly increased(P<0.01).Compared with the SPID group,the uterine index of the positive drug group and the SFF groups significantly decreased(P<0.05),the spleen index and thymus index significantly increased(P<0.05),the pathological damage of uterine tissue significantly improved,the contents of IL-1β,IL-6 and TNF-α in serum were significantly lowered(P<0.05),the contents of IgG and IgM significantly increased(P<0.05),and the TLR4,MyD88 protein and the ratio of p-NF-κB p65/NF-κB p65 protein in uterine tissue significantly downregulated(P<0.01).However,CRX-527 significantly reversed the effects of SFF on SPID in rats.Conclusion SFF could improve SPID in rats by impeding the inflammatory response,and its mechanism might be attributed to inhibition of TLR4/MyD88/NF-κB signaling pathway.

O-glycosylation(including mucin-type O-glycosylation and O-GlcNAcylation),as a critical post-translational modification(PTM),regulates protein function,stability,and subcellular localization through the addition of glycan chains to serine or threonine residues,which participates in cellular signa-ling,metabolic regulation,and stress responses.DNA damage refers to the disruption of genomic integri-ty caused by endogenous factors(e.g.,metabolic byproducts,replication errors)or exogenous agents(e.g.,radiation,chemical substances),leading to carcinogenesis,aging,and genetic disorders.To counteract DNA lesions,organisms have evolved the DNA damage response(DDR)system,which or-chestrates complex protein networks to detect DNA damage and facilitate repair processes.Emerging evi-dence indicates that O-glycosylation can modulate DDR by influencing the activity,localization,and in-teractions of DNA repair-associated proteins.However,the precise mechanisms underlying O-glycosyla-tion-mediated DDR remain to be clarified.This review systematically summarizes:(1)the biosynthetic pathways of mucin-type O-glycosylation and O-GlcNAcylation,the cascade reactions in DDR;and(2)current research advances regarding O-glycosylation in tumor-associated DDR.Furthermore,we propose novel mechanistic perspectives and therapeutic strategies targeting O-glycosylation-mediated DDR dysreg-ulation in malignancies,aiming to provide a theoretical basis for tumor treatment.

Objective:To analyze and validate the reliability and validity of the social skills improvement system-rating scales Chinese version (parent version) (SSIS-RS-C) in middle school students.Method:A total of 1 486 parents of middle school students were recruited according to the cluster sampling method.The social responsiveness scale and strengths and difficulties questionnaire were used as criterion validity tools.A retest was conducted one month later.SPSS 27.0 was used for descriptive statistics, item analysis, internal consistency test, test-retest reliability test and criterion validity test. AMOS 24.0 was used to perform confirmatory factor analysis .Results:Item analysis indicated significant positive correlations between each item and the subscales ( r=0.293-0.782, all P<0.01), with significant differences in scores between high and low groups ( t=10.079-37.038, all P<0.01).Confirmatory factor analysis supported a seven-factor structure for the social skills subscale(communication, cooperation, assertion, responsibility, empathy, engagement and self control) and a five-factor structure for the problem behavior subscale (externalizing, bullying, hyperactivity/inattention, internalizing and autism spectrum) of the SSIS-RS-C.There was a positive correlation between the social skills subscale and prosocial behavior ( r=0.637, P<0.001), and between the problem behavior subscale and social impairments and difficult behaviors ( r=0.765, 0.688, both P<0.001).The Cronbach's α coefficients for the total scale, social skills subscale and problem behavior subscale were 0.934, 0.972 and 0.963, respectively.The test-retest correlation coefficients for the total score and the two subscales were 0.665, 0.871 and 0.598, respectively (all P<0.001). Conclusion:The SSIS-RS-C demonstrated good reliability and validity in the Chinese adolescent population.

O-glycosylation(including mucin-type O-glycosylation and O-GlcNAcylation),as a critical post-translational modification(PTM),regulates protein function,stability,and subcellular localization through the addition of glycan chains to serine or threonine residues,which participates in cellular signa-ling,metabolic regulation,and stress responses.DNA damage refers to the disruption of genomic integri-ty caused by endogenous factors(e.g.,metabolic byproducts,replication errors)or exogenous agents(e.g.,radiation,chemical substances),leading to carcinogenesis,aging,and genetic disorders.To counteract DNA lesions,organisms have evolved the DNA damage response(DDR)system,which or-chestrates complex protein networks to detect DNA damage and facilitate repair processes.Emerging evi-dence indicates that O-glycosylation can modulate DDR by influencing the activity,localization,and in-teractions of DNA repair-associated proteins.However,the precise mechanisms underlying O-glycosyla-tion-mediated DDR remain to be clarified.This review systematically summarizes:(1)the biosynthetic pathways of mucin-type O-glycosylation and O-GlcNAcylation,the cascade reactions in DDR;and(2)current research advances regarding O-glycosylation in tumor-associated DDR.Furthermore,we propose novel mechanistic perspectives and therapeutic strategies targeting O-glycosylation-mediated DDR dysreg-ulation in malignancies,aiming to provide a theoretical basis for tumor treatment.

Objective To analyze the medication rules of traditional Chinese medicine in treating breast cancer based on real-world data mining.Methods Inpatients with breast cancer who received traditional Chinese medicine treatment at the Affiliated Hospital of Chengdu University of Traditional Chinese Medicine from January 2017 to December 2021 were selected.Python 3.10 software was used to mine traditional Chinese medicine prescription data;SPSS 23.0 software was applied for descriptive analysis,and systematic cluster analysis was performed on high-frequency drugs.Results A total of 3026 consultation records of inpatients with breast cancer were collected.The main traditional Chinese medicine syndrome diagnosis of"predominantly liver depression and Qi stagnation"accounted for 60.94％of the total consultations.A total of 240 kinds of traditional Chinese medicine were used,with a cumulative frequency of 35 462 times.Among them,29 kinds of traditional Chinese medicine such as Danggui,Fuling,Baizhu,Chaihu had a cumulative usage frequency exceeding 300 times.Regarding the four natures of drugs,cold-natured(43.55％),warm-natured(30.05％),and neutral-natured(23.34％)drugs were predominant;In terms of five flavors,sweet(46.12％),bitter(30.91％),and pungent(20.02％)were the main ones.The most frequently used drugs were tonifying herbs(32.77％),followed by heat-clearing herbs(15.96％)and phlegm-resolving herbs(14.71％).Systematic cluster analysis yielded 7 groups of drug combinations.Conclusion In real-world clinical practice,traditional Chinese medicine for breast cancer mainly uses tonifying herbs,reflecting the traditional Chinese medicine principle of"strengthening healthy Qi and cultivating the root"in treating tumors.The four natures and five flavors of drugs follow syndrome differentiation and the combination of cold and heat.The clustered drug combinations have extensive therapeutic effects,covering various syndromes of breast cancer at different stages,which can provide a reference for clinical medication.