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.

Objective: To explore the importance of selecting appropriate human antiserum for absorption and elution tests by analyzing two cases in which weak A antigens were detected using this method. Methods: The microcolumn agglutination and tube methods were used to perform ABO blood group typing of the patients. Absorption and elution tests were conducted to detect weak A antigens on red blood cells. Molecular biological methods were employed for genotyping. Results: Serological tests showed that the forward typing results of the two patients were O and B, respectively, and weak anti-A1 antibodies were present in their sera. When O-type human high-titer (anti-A≥256) serum was used for absorption and elution tests, patient 1 eluted anti-A, confirming the presence of weak A antigens on their red blood cells; patient 2 eluted anti-A and anti-AB simultaneously. When B-type human antiserum was used for absorption and elution, patient 2 eluted anti-A, confirming the presence of weak A antigens on their red blood cells as well. Gene sequencing results showed that the genotypes of the two patients were ABO Aw. 04/ABO O. 01. 02 and ABO AEL (c. 410C>T)/ABO B.01, respectively. Conclusion: When absorption and elution tests are needed for weak A/B antigens alone on red blood cells, O-type human high-titer serum can be prioritized. However, when both A and B antigens are present, the human antiserum selected for absorption and elution tests should only react with the weak antigen and not with the normally expressed antigen.

Objective:To investigate the effect of Xuebijing injection against blood-brain barrier(BBB)damage in the mice with anti-N-methyl-D-aspartate receptor(NMDAR)encephalitis,and to elucidate its regulatory effect on the imbalance of helper T cells 17(Th17)/regulatory T cells(Treg).Methods:The active immunization models of anti-NMDAR encephalitis in the mice were established using glutamate receptor N1 subunit(GluN1)356-385 antigen peptide,and the serum anti-NMDAR immunoglobulin G(IgG)antibody levels were detected by enzyme-linked immunosorbent assay(ELISA).The healthy mice without modeling were served as control group,and the mice with successful modeling were randomly divided into model group,low dose of Xuebijing injection(XBJ-L)group,and high dose of Xuebijing injection(XBJ-H)group,with 10 mice in each group.After modeling,the mice in XBJ-L and XBJ-H groups were intraperitoneally injected with 5 and 10 mL·kg-1 Xuebijing injection,respectively.The Longa score was used to assess the neurological impairment of the mice in various groups;evans blue(EB)staining was used to determine the BBB permeability;immunofluorescence staining was used to detect the expressions of zonula occludens 1(ZO-1)and Occludin in cerebral cortex of the mice in various groups;Western blotting method was used to determine the expression levels of ZO-1,Occludin,Claudin-5,and neuron-specific nuclear protein(NeuN)in cerebral cortex of the mice in various groups;ELISA method was used to determine the levels of Th17-and Treg-related cytokines including interleukin(IL)-17,IL-22,and IL-10 in serum of the mice;flow cytometry was used to determine the percentages of Th17 and Treg cells in peripheral blood of the mice in various groups,and the Th17/Treg ratio was calculated.Results:The serum of the mice induced with the GluN1 356-385 antigen peptide was positive for NMDAR IgG antibodies,indicating that the models were successfully established.Compared with control group,the neurological impairment score of the mice in model group was significantly increased(P＜0.05),and the EB level in brain tissue was significantly increased(P＜0.05);the fluorescence staining intensities of ZO-1 and Occludin in the cerebral cortex were decreased,and the expression levels of ZO-1,Occludin,Claudin-5,and NeuN proteins in the cerebral cortex were significantly decreased(P＜0.05);the serum levels of IL-17 and IL-22 were significantly increased(P＜0.05),while the IL-10 level was significantly decreased(P＜0.05);the percentage of Th17 cells in peripheral blood was significantly increased(P＜0.05),while the percentage of Treg cells was significantly decreased(P＜0.05),and the Th17/Treg ratio was significantly increased(P＜0.05).Compared with model group,the neurological impairment scores of the mice in XBJ-L and XBJ-H groups were significantly decreased(P＜0.05),the EB levels in brain tissue were significantly decreased(P＜0.05),the fluorescence staining intensities of ZO-1 and Occludin in cerebral cortex were increased,and the expression levels of ZO-1,Occludin,Claudin-5,and NeuN proteins were significantly increased(P＜0.05);the levels of IL-17 and IL-22 in serum were significantly decreased(P＜0.05),and the level of IL-10 was significantly increased(P＜0.05);the percentages of Th17 cells in peripheral blood were significantly decreased(P＜0.05),the percentages of Treg cells were significantly increased(P＜0.05),and the Th17/Treg ratios were significantly decreased(P＜0.05).Compared with XBJ-L group,the neurological function injury score of the mice in XBJ-H group was significantly decreased(P＜0.05),the EB level in brain tissue was significantly decreased(P＜0.05);the fluorescence staining intensities of ZO-1 and Occludin in the cerebral cortex were increased,and the expression levels of ZO-1,Occludin,Claudin-5,and NeuN proteins were significantly increased(P＜0.05);the serum levels of IL-17 and IL-22 were significantly decreased(P＜0.05),and the level of IL-10 was significantly increased(P＜0.05);the percentage of Th17 cells in peripheral blood was significantly decreased(P＜0.05),the percentage of Treg cells was significantly increased(P＜0.05),and the Th17/Treg ratio was significantly decreased(P＜0.05).Conclusion:Xuebijing injection can improve BBB injury,regulate Th17/Treg balance,and thereby alleviate the neurological functional damage in anti-NMDAR encephalitis.

Efferocytosis refers to the process of phagocytes engulfing and clearing the cells after programmed cell death. In recent years, an increasing number of studies have shown that the mechanisms of efferocytosis are closely related to drug-induced liver injury, hepatic ischemia-reperfusion injury, viral hepatitis, cholestatic liver diseases, metabolic-associated fatty liver disease, alcoholic liver disease, and other liver disorders. This review summarized the research progress on the role of efferocytosis in liver diseases, with the hope of providing new targets for the prevention and treatment of liver diseases.
Humans ; Liver Diseases/metabolism* ; Animals ; Phagocytosis/physiology* ; Phagocytes ; Efferocytosis

Objective:To observe the clinical progression of low-risk papillary thyroid microcarcinoma（LR-PTMC）, analyze the influencing factors of its oncological outcomes, and explore the feasibility of active surveillance（AS） of LR-PTMC. Methods:This study adopted a prospective observational research design. A total of 85 subjects diagnosed with LR-PTMC during health checkup in Health Management Center of our hospital from March 2021 to October 2022 were enrolled as the research subjects, for at least 2 years of AS follow-up observation. The clinical progress and oncological outcomes were recorded, disease progression was defined as any increase in nodule diameter ≥3 mm or the appearance of new lesions or lymph node metastasis or distant metastasis, and the oncological outcome was use disease progression defining. Cox proportional hazards regression model was used to analyze the influencing factors of oncological outcomes in LR-PTMC patients. Results:A total of 85 LR-PTMC patients who underwent physical examinations were included in this study. The median follow-up time was 2 years, and a total of 23 patients（27.06%） experienced disease progression. Among them, 18 patients（21.18%） had enlarged lesions（any nodule diameter increased by ≥3 mm）, and 5 patients（5.88%） had abnormal or metastatic cervical lymph nodes. The 2-year cumulative disease progression rate was 9.41%. The incidence age of LR-PTMC patients was younger, with a higher proportion of ≤45 years old. The proportion of baseline nodules with a maximum diameter greater than 5 mm is higher, and the proportion of baseline TPO Ab positivity was higher. Ultrasound showed a higher proportion of microcalcifications compared to the non progression group, and the differences were statistically significant（all P<0.05）. Multivariate Cox proportional hazards regression analysis showed that age of onset ≤45 years RR 95% CI 1.052（1.018-1.088） and ultrasound showing microcalcifications RR 95% CI 3.361（1.379-8.194） were independent risk factors affecting disease progression during AS in LR-PTMC patients（P<0.05）. Conclusion:Most LR-PTMC patients maintain stable lesion size and low lymph node metastasis rate during the AS process, with good oncological outcomes in the short term. AS can be considered as a safe and effective alternative to surgical treatment for LR-PTMC patients. But for patients with onset age ≤45 years and microcalcifications, the follow-up interval can be shortened for close observation.
Humans ; Thyroid Neoplasms/pathology* ; Disease Progression ; Prospective Studies ; Carcinoma, Papillary/pathology* ; Female ; Male ; Middle Aged ; Adult ; Watchful Waiting ; Lymphatic Metastasis ; Proportional Hazards Models ; Risk Factors

Traditional development of small protein scaffolds has relied on display technologies and mutation-based engineering, which limit sequence and functional diversity, thereby constraining their therapeutic and application potential. Protein design tools have significantly advanced the creation of novel protein sequences, structures, and functions. However, further improvements in design strategies are still needed to more efficiently optimize the functional performance of protein-based drugs and enhance their druggability. Here, we extended an evolution-based design protocol to create a novel minibinder, BindHer, against the human epidermal growth factor receptor 2 (HER2). It not only exhibits super stability and binding selectivity but also demonstrates remarkable properties in tissue specificity. Radiolabeling experiments with ^99mTc, ⁶⁸Ga, and ¹⁸F revealed that BindHer efficiently targets tumors in HER2-positive breast cancer mouse models, with minimal nonspecific liver absorption, outperforming scaffolds designed through traditional engineering. These findings highlight a new rational approach to automated protein design, offering significant potential for large-scale applications in therapeutic mini-protein development.

In recent years, with the endless emergence of meibomian gland dysfunction(MGD)diagnostic equipment, rich treatment methods, and in-depth clinical and basic research on MGD at home and abroad, the understanding of MGD has entered a new stage. MGD-related dry eye is considered to be the main cause of lipid abnormal dry eye, and its occurrence and development is a chronic and multi-factorial pathological process. This article reviews the pathogenesis, imaging analysis and clinical treatment progress of MGD-related dry eye, in order to provide scientific evidence and ideas for clinical diagnosis and therapy of MGD-related dry eye.

Digital health technologies (DHT) not only enhance patient engagement in healthcare but also enable personalized treatment and care services, empowering patients to manage their own health well. This paper reviews the composition and characteristics of digital vulnerable groups, as well as the current application status, influencing factors, intervention measures, and insights of DHT in their care. This paper aims to provide theoretical and practical references for enhancing the quality of DHT care for digital vulnerable groups, thereby promoting targeted and effective development of DHT within the nursing field.

Purpose To investigate the value of peritumoral expansion-based radiomics from tumor regions of interest(ROIs)for predicting epidermal growth factor receptor(EGFR)mutation status and to identify the optimal peritumoral expansion margin.Materials and Methods This retrospective study included 390 patients with pathologically confirmed non-small cell lung cancer(NSCLC)and definitive EGFR genotyping results from the Affiliated Huai'an No.1 People's Hospital of Nanjing Medical University(December 2021 to September 2023).Patients were randomly divided into training and validation sets(8∶2 ratio,310 vs.80 patients;CT slice thickness:5-7 mm).An additional independent test set of 100 patients undergoing thin-section CT(1-2 mm)was included to assess model generalizability across slice thicknesses.Clinical characteristics and CT semantic features were analyzed.After automated tumor segmentation with manual refinement,ROIs were sequentially expanded outward by 1,2,3,4 and 5 mm.A total of 2 264 radiomic features were extracted from each ROI.Feature selection using minimum redundancy maximum relevance and least absolute shrinkage and selection operator algorithms was performed in the training set to calculate radiomics scores.Logistic regression was used to develop prediction models.A combined model integrating optimal peritumoral radiomics score with clinical and CT features was established,with performance compared across models.Results Gender(χ2=24.922,P<0.001),smoking history(χ2=11.199,P=0.001),emphysema(χ2=40.802,P<0.001),and lymph node metastasis(χ2=5.674,P=0.017)were associated with EGFR mutation status.In the validation set,the area under the curve for the tumor ROI-based radiomics model was 0.676,while the five expansion models achieved area under the curve of 0.723,0.720,0.734,0.681,and 0.598,respectively.The combined model based on 3 mm expansion demonstrated superior performance to individual radiomics and clinical models,with area under the curve of 0.826,0.734,0.711 in the validation set,and 0.809,0.760,0.702 in the independent test set.Conclusion Peritumoral expansion-based CT radiomics demonstrates value for predicting EGFR mutations in NSCLC,with 3 mm identified as the optimal expansion margin.Integration with clinical information further enhances predictive accuracy.

Objective:To construct a debriefing and reflection training program for nursing undergraduate interns, and to provide a basis for implementing such a program and improve the practical clinical skills of the interns.Methods:Based on competency theory, the draft of the debriefing and reflection training program for undergraduate nursing interns was constructed through literature review and expert conference discussion. Delphi expert consultation was conducted from August to October 2024, and the final version of the program was established based on the experts' opinions. The weight of each indicator was determined using the analytic hierarchy process. Excel 2019 was used for data entry, and SPSS 19.0 was used for data analysis.Results:Two rounds of expert consultation were conducted, with a questionnaire recovery rate of 100.00% for both. The experts consulted in the second round had an authority coefficient of 0.861, a coefficient of variation for each index of 0.048 to 0.237, and a Kendall's concordance coefficient of 0.137 ( P<0.05). The final program included 5 primary indicators, 16 secondary indicators, and 73 tertiary indicators. Conclusions:The debriefing and reflection training program for undergraduate nursing interns proposed in this study has high scientifical validity and reliability. It can provide a reference for debriefing and reflection training of nursing undergraduate interns in China, thereby cultivating qualified clinical nursing talents.