To develop a prediction model for placenta previa in early pregnancy by analyzing embryo implantation sites and related clinical factors, in order to provide an objective basis for early risk identification.

A retrospective cohort study was conducted on 232 singleton pregnant women delivering in our hospital between September 2020 and March 2024. According to the final pre-delivery ultrasound findings, they were divided into placenta previa group (n=78) and a non-placenta previa group (n=154). Their ultrasound parameters [distance from lower gestational sac margin to cervical os, implantation site (lower/middle-upper uterine segment), and implantation position (anterior/posterior wall)], and clinical data [age, gravidity (categorized as <2 or ≥2 pregnancies), parity (<2 or ≥2 deliveries), and history of intrauterine procedures were collected through electronic medical records. Univariate analysis was used to screen potential predictors (P<0.1), and multivariate logistic regression analysis was employed to identify the predictors (P<0.05) for placenta previa. Then a nomogram prediction model was constructed, which was internally validated with Bootstrap (1 000 bootstrap resamples) and assessed for discrimination with area under the receiver operating characteristic curve (AUC) and for calibration with Hosmer-Lemeshow goodness-of-fit test.

The placenta previa group showed significantly advanced age (≥35 years), lower education level (≤high school), multigravidity (≥2 pregnancies), multiparity (≥2 deliveries), more intrauterine procedures, distance of gestational sac-to-cervical os <5.5 mm, and larger proportion of lower uterine segment implantation than the non-placenta previa group (all P<0.1). But there were no statistical differences between the 2 groups in proportion of posterior wall implantation, history of cesarean section or assisted reproductive technology (ART). Multivariate logistic regression analysis confirmed lower uterine segment implantation (OR=40.40, 95%CI: 14.68~136.19, P<0.001), posterior wall implantation (OR=2.73, 95%CI:1.27~6.28, P=0.013), and intrauterine procedures (OR=3.48, 95%CI: 1.65~7.70, P=0.001) as independent risk factors. The model based on these predictors demonstrated excellent discrimination (AUC value=0.84, 95%CI: 0.79~0.90) and calibration (Hosmer-Lemeshow test Chi-square=3.455, P=0.750).

Lower uterine segment/posterior wall implantation and intrauterine procedures are independent risk factors for placenta previa. Our nomogram model based on these factors shows good predictive efficiency, and can provide reference for early recognition of pregnant woman with high-risk placenta previa.

To develop and validate a deep learning model based on chest CT images to accurately distinguish between drug-resistant (DR-TB) and -sensitive tuberculosis (DS-TB).

A retrospective study was conducted on 722 cases of confirmed secondary tuberculosis admitted in our center from January 2019 to December 2022. According to the results of antimicrobial susceptibility test, they were divided into 357 DS-TB cases and 365 DR-TB cases. Pre-existing U-Net segmentation model was employed to segment the lung parenchyma regions in CT images. The dataset was randomly partitioned into a training set and a testing set in an 8:2 ratio. Six 3D deep learning architectures (3D Swin Transformer, 3D ShuffleNet v2, 3D ViT, 3D MobileNet v2, 3D DenseNet, and 3D ResNet18) were employed to evaluate the discriminative efficiency between DS-TB and DR-TB. Hyperparameters were optimized by five-fold cross-validation on the training set to construct the optimal model. The performance of the constructed model was assessed using area under the curve (AUC), sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and F1-score. Six radiologists independently evaluated DR-TB identification on the test set, and their performance was compared with the best-performing deep learning model.

The AUC value in DR-TB prediction was 0.583, 0.704, 0.698, 0.758, 0.736, and 0.841, respectively, for 3D Swin Transformer, 3D ShuffleNet v2, 3D ViT, 3D MobileNet v2, 3D DenseNet, and 3D ResNet18. The 3D ResNet18 model demonstrated optimal performance, achieving a sensitivity of 0.935 (95%CI: 0.880~0.987), a specificity of 0.642 (95%CI: 0.492~0.757), a PPV of 0.750 (95%CI: 0.663~0.835), an NPV of 0.896 (95%CI: 0.809~0.976), an AUC value of 0.841, and a F1-score of 0.832. The radiologists got a F1-score of 0.571, 0.450, 0.675, 0.623, 0.617 and 0.635, respectively, and the F1-score of the 3D ResNet18 model is all higher than that of the radiologists. The highest-performing radiologist achieved sensitivity, specificity, PPV and NPV of 0.701 (95%CI: 0.605~0.802), 0.567 (95%CI: 0.447~0.684), 0.651 (95%CI: 0.549~0.757), and 0.623 (95%CI: 0.500~0.754), with all these values lower than those of the 3D ResNet18 model (P<0.05). Class activation mapping showed that the 3D ResNet18 model could focus on key lesion areas. The class activation mapping demonstrated that the 3D ResNet18 model could effectively focus on critical lesion regions.

Our 3D ResNet18 model shows the best predictive performance in identifying DR-TB, and is expected to assist clinical diagnosis for DR-TB.

To investigate the protective effects of Qiji Special Dietary Food on exercise-induced myocardial injury in mice.

An exercise-induced myocardial injury model was established using a treadmill running protocol, and at the same time, the modeled mice were administered Qiji Special Dietary Food via oral gavage, followed by a 4-week treadmill exhaustion test. Serum levels of cardiac troponin T (cTnT), creatine kinase (CK), and lactate dehydrogenase (LDH) were measured. Myocardial tissues were analyzed for superoxide dismutase (SOD) activity and malondialdehyde (MDA) content. Histopathological alterations and ultrastructural changes in myocardial tissue were evaluated using light microscopy and transmission electron microscopy (TEM).

After 4 weeks of exhaustive training, compared to the control group, the model group exhibited significantly elevated serum cTnT, CK, and myocardial MDA levels (P<0.01), along with reduced myocardial SOD activity (P<0.01). Compared to the model group, high-, medium-, and low-dose treatment significantly attenuated the exhaustive exercise-induced increases in serum cTnT level and myocardial MDA content (P<0.01), restoring these indicators to the levels comparable to those of the normal control group (P<0.05). Additionally, all treatment groups had markedly increased myocardial SOD activity, with no significant difference from the normal group. Histopathological and ultrastructural analyses revealed markedly alleviated myocardial damage in the treatment groups, with the medium-dose group exhibiting the most pronounced protective effects.

Qiji Special Dietary Food demonstrates significant protective effects against exercise-induced myocardial injury in mice, which maybe associated with its antioxidant activity and mitigation of oxidative stress.

To investigate whether olfactory mucosa mesenchymal stem cells (OM-MSCs) attenuate oxygen glucose deprivation and recovery (OGD/R)-induced ferroptosis in neurons through glutathione oxidase 4 (GPX4).

The middle nasal tissue were collected from a patient with nasal polyps admitted in our hospital, and then OM-MSCs were isolated from the tissue, which were confirmed by morphological observation under light microscopy and phenotypic characterization through flow cytometry for surface markers, including CD34, CD45, CD73, CD90, CD105, and CD146. Mouse hippocampal neuronal cell line HT22 was randomly divided into control, Control, OGD/R, OGD/R+OM-MSCs, OGD/R+sh-NC, OGD/R+sh-GPX4 and OGD/R+sh-GPX4+OM-MSCs groups. After the cells were subjected to OGD/R modeling, the cells were subsequently co-cultured with OM-MSCs and/or knockdown of GPX4. Neuronal apoptosis was quantified by flow cytometry, while cell viability was assessed using CCK-8 assay. Biochemical markers associated with ferroptosis, including MDA, ROS, GSH, and Fe²⁺ levels, were measured with corresponding reagent kits. The GPX4 expression at both mRNA and protein levels was determined through qPCR and Western blotting, respectively.

The isolated and primarily cultured OM-MSCs showed typical characteristics of OM-MSCs in cell surface markers (negative expression of CD34 and CD45 but positive expression of CD73, CD90, CD105, and CD146 on cell surface) and morphology (adherent cells in a spindle-like shape). Significant differences were observed among the control, OGD/R, and OGD/R+OM-MSCs groups in terms of cell viability, MDA, ROS, GSH, Fe²⁺ and GPX4 (P<0.05). The OGD/R group showed notable decreases in cell activity and GSH (P<0.05), increases in MDA, ROS, and Fe²⁺ (P<0.05), and down-regulation of GPX4 when compared with the control group (P<005). Co-culture with OM-MSCs enhanced cell activity and GSH (P<0.05), decreased MDA, ROS, and Fe²⁺ (P<0.05), and up-regulated GPX4 as compared to the conditions in the OGD/R group (P<0.05). While, OGD/R+sh-GPX4 treament developed the decreases in cell viability, GSH, and GPX4 and the increases in MDA, ROS, and Fe²⁺ as compared to the OGD/R+sh-NC group (P<0.05), however, all of these could be reversed by OM-MSCs.

OM-MSCs inhibit OGD/R-induced ferroptosis in HT22 cells by up-regulating GPX4.

To investigate the effects of open reading frame 1 protein (ORF1p), encoded by long interspersed nuclear element-1 (LINE-1), on the proliferation, migration, and invasion of esophageal squamous cell carcinoma (ESCC) cells, and explore the underlying molecular mechanism.

① Western blotting was performed to compare the expression of ORF1p between normal esophageal squamous epithelial cells and ESCC cells. ② Immunohistochemistry (IHC) assay was used to examine ORF1p expression in ESCC tissues and paired normal tissues adjacent to tumor. ③ The effects of ORF1p knockdown and overexpression on malignant behaviors in ESCC cells were determined through functional assays. ④ Xenograft tumor model in nude mice was established to evaluate the impact of ORF1p on tumor growth in vivo. ⑤ Transcriptome sequencing combined with cell functional rescue experiments were conducted to identify downstream targets regulated by ORF1p.

① Western blot analysis demonstrated the expression of ORF1p was significantly higher in the ESCC cell lines than the normal esophageal squamous epithelial cells (P<0.05). ② IHC confirmed remarkable up-regulation of ORF1p in ESCC tissues than paired adjacent normal tissues (P<0.000 1). ③ Functional assays and experiments on xenograft tumor models revealed that ORF1p substantially enhanced the proliferation, migration, and invasion of ESCC cells, as well as tumorigenic potential in vivo (P<0.05). ④ Functional rescue experiments showed that ORF1p facilitated the proliferation, migration, and invasion of ESCC cells by modulating AJUBA expression (P<0.05).

ORF1p is significantly up-regulated in ESCC and promotes the proliferation, migration, and invasion of ESCC cells by regulating AJUBA expression.

To investigate the role and primary mechanism of a novel fusion gene RELCH-RET in driving the malignant transformation of normal human bronchial epithelial (HBE) cells.

Based on retrospective clinical data from 456 non-small cell lung cancer (NSCLC) patients admitted in the Second Affiliated Hospital of Army Medical University from January 2019 to June 2022, a fusion gene, RELCH-RET,was identified as a research target. Three cell models were established: negative control (HBE VC, transfected with empty lentiviral vector), RET control (HBE RET, transfected withlentiviral overexpression vector of Flag-RET), and experimental group (HBE RELCH-RET, transfected with lentiviral overexpression vector of Flag-RELCH-RET). MTS assay and Transwell assay were used to detect cell proliferation and migratory and invasive abilities. In vivo tumorigenicity of the 3 cell models was assessed in 15 female non-obese diabetic/severe combined immunodeficiency (NOD/SCID) mice (SPF grade, 4 weeks old, weighing 15.1±0.4 g) via subcutaneous xenograft experiments, with 5 animals in each group. Western blotting was employed to detect the autophosphorylation of RET (Y905) and the phosphorylation of downstream signaling proteins ERK1/2, EGFR (Y845) and STAT3 (Y705). Dimerization and multimerization status of RELCH-RET were analyzed by chemical cross-linking (DTME treatment) in combination with Western blotting, with the reversibility being confirmed through de-cross-linking experiments.

There were 3 cases carrying RELCH-RET fusion gene screened out from the 469 NSCLC patients. Compared with the HBE VC and HBE RET groups, the HBE RELCH-RET group exhibited significantly enhanced cell proliferation (P<0.01), and acquired migratory and invasive abilities (P<0.01), while the control groups did not demonstrate the abilities. In the mouse xenograft tumor model, HBE cells stably expressing RELCH-RET developed significant tumor nodules (P<0.001), whereas the control groups (empty vector and wild-type RET) failed to exhibit detectable tumor growth. Western blotting revealed that RELCH-RET could induce the autophosphorylation of the RET tyrosine residue (Y905) and significantly up-regulate the phosphorylation levels of ERK1/2, EGFR (Y845), and STAT3 (Y705) proteins. Chemical cross-linking combined with Western blot analysis demonstrated that RELCH-RET formed a dimer (~170 kDa) in HBE cells, which is reversibly dissociated into monomers upon decross-linking treatment.

The novel fusion gene RELCH-RET, promotes ligand-independent dimerization/oligomerization, thereby mediating RET autophosphorylation, subsequently activates the downstream typical RET signaling pathway and ultimately drives the malignant transformation of normal HBE cells.

To investigate the effect of nutritional deficiency on autophagic flux in human hepatoblastoma HepG2 cells and the autophagy-dependent molecular pathway mediated by ATG5 against nutritional stress.

Lentiviral siRNA knockdown of ATG5 was performed to knock down the expression in HepG2 cells, and the transfection was verified by Western blotting and qRT-PCR. HBSS was used to treat HepG2 cells for 0, 1, 2, 3, 4, 5, 6 and 7 h for starvation-induced autophagy. Monodansylcadaverine (MDC) fluorescence staining was employed to detect the formation of intracellular autophagic vesicles. Western blotting was performed to measure the expression changes in microtubule-associated protein 1A/1B-light chain 3 (LC3), autophagy receptor protein (sequestosome 1, P62), autophagy-related gene 5 (ATG5) and nutrient stress pathway (AMPK/mTOR pathway) related proteins. CCK-8 assay was utilized to test cell proliferation.

Compared with the HepG2 cells under conventional culture medium, starvation induction resulted in more autophagic vacuoles (accumulation of autofluorescent marker, MDC), which peaked at 6 h of starvation (P<0.000 1), up-regulated protein levels ofLC3B-Ⅱ and ATG5 (P<0.05), decreased protein expression ofP62 (P<0.05), gradual activation of the AMPK/mTOR pathway with elapse of starvation time, and increase in the intensity of autophagic flux. However, in the HepG2 cells with ATG5 knockdown, starvation treatment led to decreased ATG5 expression, no significant difference in LC3B-II and P62 changes, inhibited autophagic flux, and suppressed cell proliferation (P<0.000 1).

Human hepatoblastoma HepG2 cells can regulate autophagic flux to against nutritional stress by inducing ATG5 expression.

To investigate the effect of endothelial cells in the perivascular niche on the stem cell-like characteristics of osteosarcoma cells and primarily explore the possible molecular mechanism.

A co-culture model was established in vitro using SV40T- human umbilical vein endothelial cells (HUVEC-T1) and osteosarcoma stem cells (OSCs) derived from the human osteosarcoma cell line 143B.Thus, there were 2 groups of cells, OSCs and co-cultured OSCs. The self-renewal capacity of OSCs between the 2 groups was assessed using a limiting dilution forming sphere assay. Flow cytometry and Western blotting were used to detect the expression of stem cell marker CD133 and stemness transcription factors SOX2 and NANOG. Fourteen female nude mice (4~6 weeks old, weighing 18~20 g) were randomly and equally divided into 2 groups of subcutaneous xenograft models: the control group (OSCs suspension) and the experimental group (OSCs+HUVEC-T1 mixed suspension). The tumor volume and mass were compared between the 2 xenograft groups. Immunofluorescence (IF) staining was used to verify the spatial proximity between endothelial cells and OSCs in vivo, while immunohistochemistry was employed to compare microvessel density (MVD) and CD133 expression level between the 2 groups. RNA-seq was performed to identify potential signaling pathways of endothelial cells affecting the stemness of OSCs. PCR and Western blotting were applied to confirm the RNA-seq findings. Exogenous protein treatment, IF staining, Western blotting and sphere formation assay were utilized to preliminarily validate the role of the identified pathway in regulating the stemness phenotype of OSCs.

The in vitro co-culture model of HUVEC-T1 and OSCs was successfully established. Compared with the control group, the co-culture group exhibited significantly enhanced self-renewal ability of OSCs, laeger proportion of the stemness marker CD133⁺ [(8.20±1.64)% vs (4.32±1.34)%, P<0.05], enhanced expression of CD117, SOX2 and NANOG (P<0.05), along with more sphere formation (P<0.05) and elevated SOX2/NANOG protein levels. The xenograft mice from the experimental group showed larger tumor volume (643.10±413.50 vs 247.90±93.66 mm³, P<0.05) and heavier tumor weight (0.52±0.27 vs 0.24±0.10 g, P<0.05) when compared the control group, correspondingly showing increased MVD (22.57±11.84 vs 11.43±5.38, P<0.05) and elevated CD133 expression (P<0.05). IF staining confirmed the adjacency of CD31-labeled endothelial cells and CD133-labeled OSCs in vivo. RNA-seq and functional experiments demonstrated that CXCR4 was highly expressed in co-cultured OSCs, CXCL12 was highly expressed in co-cultured endothelial cells, and exogenous CXCL12 promoted the sphere formation and expression levels of SOX2 and NANOG of OSCs (P<0.05).

Endothelial cells within the perivascular niche may promote the stemness phenotype of osteosarcoma cells via the CXCL12/CXCR4 pathway.

To investigate the effect and underlying mechanism of serine and arginine rich splicing factor 1 (SRSF1) on the replication of hepatitis B virus (HBV).

The effects of SRSF1 on HBV replication were investigated in different HBV replicating cell models by Southern blotting, Northern blotting and ELISA. Quantitative PCR , luciferase reporter assay and chromatin immunoprecipitation (ChIP) were applied to determine the effects of SRSF1 on the activities of HBV core promoter/enhancer in HepG2 cells. The relationship between SRSF1 and P53 was explored with Western blotting and ubiquitination assay. The effects of P53 on HBV replication were verified in different HBV replicating cell models, and the role of P53 in SRSF1 inhibition of HBV was clarified.

Overexpression of SRSF1 significantly inhibited HBV DNA and HBV RNA and reduced HBsAg and HBeAg secretion levels in a variety of HBV replicative cell models (P<0.0001). SRSF1 also inhibited the activity of HBV core promoter, although this inhibition was regulated by indirect mechanisms. In addition, SRSF1 enhanced P53 stability by protecting P53 from ubiquitination and subsequent proteasomal degradation. Meanwhile, the regulatory effect of overexpression or knockdown of P53 on HBV was validated in different cell models (P<0.0001).

Overexpression of splicing factor SRSF1 significantly inhibits HBV replication in a variety of cell models, and this inhibitory effect is mediated by its enhancement of P53 stability.

To investigate the clinical significance of the miR-543/syntrophin beta 1 (SNTB1) axis in colorectal cancer (CRC) and its influence on the tumor immune microenvironment.

The expression of SNTB1 in CRC tissues was analyzed using public data, such as, The Cancer Genome Atlas (TCGA), Genotype-Tissue Expression (GTEx), and Human Protein Atlas (HPA). Then Kaplan-Meier survival analysis, univariate Cox regression analysis and correlation analysis were performed to evaluate the prognostic value of SNTB1 and its relationship with immune microenvironment in CRC. The targeting relationship between miR-543 and SNTB1 was confirmed through online databases and fluorescence assays in HT-29 cells. For in vitro experiments, after transfecting si-SNTB1, miR-543 mimics and/or SNTB1 overexpression plasmids, HT-29 cells were co-cultured with CD8⁺ T cells, the expression of miR543 and SNTB1 and the viability and cytotoxicity of CD8⁺ T cells were assessed with qRT-PCR, Western blotting, flow cytometry, ELISA, and lactate dehydrogenase (LDH) release assay.

Analysis of public databases revealed significantly higher expression of SNTB1 in CRC tissues than normal tissues (P<0.001). The CRC patients with high SNTB1 expression exhibited poorer prognosis when compared with those with low expression level (P<0.05). Moreover, high SNTB1 expression was negatively correlated with immune scores in the tumor microenvironment and immune cell infiltration, especially CD8⁺ T cells (P<0.05). Furthermore, Knockdown of SNTB1 in HT-29 cells enhanced the cytotoxic activity of CD8⁺ T cells (P<0.01). Online database and in vitro experiments confirmed that miR-543 targets SNTB1, while the expression of miR-543 was decreased in colorectal cancer (P<0.001). Transfection with the miR-543 mimic inhibited the expression of SNTB1 in HT-29 cells (P<0.001), while overexpressing SNTB1 counteracted the promotion effect of miR-543 mimics on CD8⁺ T cell-mediated cytotoxicity (P<0.05).

MiR-543 activates CD8⁺ T cells and enhances their cytotoxicity against HT-29 cells by directly targeting SNTB1.