To explore the prevention effectiveness of a mode, conducting targeted corrective training based on motor dysfunction detected by military joint function screening, on military training injuries in new recruits during recruit basic training in order to cope with the high incidence of military training injuries among them.

A military personnel joint function screening was conducted on the new recruits in a training base of Joint Logistics Support Force. Based on the results of screening, the new recruits with a single action score of 1 and a total score of <10 were subjected and served as corrective training participants. Through cluster sampling, the new recruits were randomly divided into an experimental group (n=223) and a control group (n=223). The control group were trained according to the regular training plan, while the experimental group completed their designated training tasks and a 2-month corrective training for motor dysfunction at the same time. The entire corrective training process was carried out by our key personnel who had received relevant training. Medical records of the medical security department of the experimental unit were collected and evaluated for the injury situation. The score of joint function screening was analyzed using independent sample t test. The incidence of training injuries was analyzed using Chi-square test or Fisher’s exact test.

After corrective training, the score of joint function was higher in the experimental group than the score before training (15.12±2.13 vs 10.58±2.83), and the score was also higher than that of the control group (15.12±2.13 vs 14.19±1.97, P<0.05). During the recruit basic training period, the incidence of training injuries was notably lower in the experimental group than the control group (12.5% vs 34.5%, Chi-square=5.469, P=0.001). Only for those who were injured during the training, 11 people (39.2%) in the experimental group scored less than 10, which was obviously lower than the 49 people (63.6%) in the control group (Chi-square=4.972, P=0.026).

Our mode, corrective training based on the results of military personnel joint function screening, can effectively reduce the incidence of military training injuries in new recruits during recruit basic training, and exerts a good preventive effect against military training injuries.

To determine the acute effects of blood flow restriction (BFR) running warm-up on Achilles tendon morphology and function in basketball players in order to provide a theoretical basis for optimizing warm-up protocols for military personnel and athletes susceptible to Achilles tendon injuries.

Twenty-seven male basketball players were subjected and asked to participate in 3 different running warm-up protocols: low-speed running (LSR), high-speed running (HSR), and BFR combined with LSR (BFR-LSR). The acute changes in Achilles tendon morphology, mechanical properties, and functional performance across the 3 testing sessions were analyzed and compared.

Immediately after training, both HSR warm-up and BFR-LSR warm-up significantly improved Achilles tendon thickness, blood flow, stiffness, and gastrocnemius maximal voluntary isometric contraction (MVIC) when compared with LSR warm-up (P<0.05). No statistical differences were observed in above indicators between the BFR-LSR and HSR warm-ups (P>0.05). 24 hours after training, compared with LSR warm-up, HSR warm-up still significantly improved Achilles tendon thickness, blood flow, stiffness, and gastrocnemius MVIC (P<0.05). Although BFR-LSR warm-up did not show statistically significant differences in these parameters compared to LSR warm-up, it still demonstrated positive trends. Immediately and 24 h after training, no obvious difference were found in jump performance among the 3 warm-up protocols (P>0.05), but, both BFR-LSR and HSR warm-ups exhibited superior performance than LSR warm-up.

Immediately after training, BFR-LSR warm-up demonstrates comparable effects to the HSR warm-up on improving Achilles tendon morphology and performance, as well as enhancing jump performance. However, its sustained and long-term effects require further investigation.

To explore the effect of rotenone exposure on the metabolic homeostasis of nicotinamide adenine dinucleotide (NAD⁺) in dopaminergic neurons of the rat mid-brain striatum, and investigate the effect of exogenous NAD⁺ intervention on the cellular damage response of dopaminergic neurons induced by rotenone.

Male SD rats (8 weeks old, 200~250 g) were divided into a control group using a table of random numbers, a rotenone exposure group, an NAD⁺-intervention group, and an NAD⁺ group. An intoxication model was established in the rotenone exposure group. NAD⁺ (250 mg/kg) was administered simultaneously with rotenone exposure in the NAD⁺ -intervention group. The NAD⁺ group was only given NAD⁺, while the control group received no intervention. After modeling, open field test was performed to evaluate behavioral changes. After scarification, serum samples and mid-brain striatal tissues were collected. HE staining was used to observe the morphology of dopaminergic neurons in the striatum. The NAD⁺ content in the tissues was detected with NAD⁺/NADH kit. Western blotting was employed to determine the contents of tyrosine hydroxylase (TH), nicotinamide phosphoribosyltransferase (NAMPT), nicotinamide mononucleotide adenylyltransferase (NMNAT), and solute carrier family 25 member A51 (SLC25A51). ELISA was utilized to measure the content of dopamine in the striatal tissues. Immunohistochemical staining was applied to observe the distribution and contents of TH proteins in the striatal tissues of each group.

Rotenone exposure significantly affected the vital signs and motor abilities of rats, induced disorderly-arranged, atrophy and deformed neurons in the striatal tissue, decreased the content of TH, rate-limiting enzyme for dopamine synthesis, by approximately 29% (P<0.01), the content of dopamine by about 42%, and that of NAD⁺ by almost 50% (P<0.01), while increased the NADH/NAD⁺ ratio (P<0.01). After exposure, the content of NAMPT, an enzyme related to NAD⁺ synthesis, was decreased by 26% (P<0.05), the contents of NMNAT1-3 and SLC25A51, mitochondrial transporters of NAD⁺ by approximately 21%, 38%, 43%, and 21%, respectively (P<0.01). Exogenous NAD⁺ intervention improved the motor function of exposure rats and the morphology of dopaminergic neurons in the mid-brain striatal tissue, and restored the content of TH in the striatal tissue significantly by 12.8% (P<0.05), and the content of dopamine by 20.9% (P<0.05).

Rotenone disrupts the NAD⁺ homeostasis in dopaminergic neurons by inhibiting the NAD⁺ synthesis and transport pathways in the mid-brain striatal tissues, while exogenous NAD⁺ intervention can effectively alleviate the dopaminergic neuron damage induced by rotenone exposure.

To explore the underlying mechanisms by which time-restricted feeding (TRF) attenuates dextran sodium sulfate (DSS)-induced colitis in mice via modulation of regenerating islet-derived protein 3 gamma(Reg3γ) expression and gut microbiota.

Six-week-old C57BL/6 mice were stratified by body weight and randomized into ad libitum feeding (AL) and TRF groups (n=7). The AL mice were given unrestricted food access, whereas the TRF mice were allowed feeding only during 00:00 and 08:00 daily, for totally 4 weeks. Mouse colitis model was induced at the fourth week by adding 2.5% dextran sodium sulfate (DSS) in drinking water for 6 d. Disease severity and the effects of TRF were assessed with disease activity index (DAI) scoring, colon length measurement, HE staining and histopathological scoring, and mRNA expression levels of regenerating islet-derived 3 gamma（Reg3g） and inflammatory cytokines in colonic tissues. Another 14 mice were randomized into AL plus antibiotic cocktail (AL+ABX) and TRF plus antibiotic cocktail (TRF+ABX) groups, with 7 animals in each group. ABX was administered to deplete gut microbiota and evaluate the microbiota dependence of TRF in attenuating colitis. Fecal samples from AL and TRF mice were analyzed by 16S ribosomal RNA sequencing (16S rRNA-seq), and serum lipopolysaccharide (LPS) level was measured. The colonic epithelial cells were collected for RNA-seq.

After modeling, the AL mice exhibited typical colitis symptoms, such as weight loss, diarrhea, and hematochezia. TRF intervention significantly attenuated these above symptoms, with lower DAI scores from day 4 post-modeling (P<0.001), reduced colon shortening (P<0.01), preserved tissue architecture, and decreased inflammation. RT-qPCR analysis showed that TRF down-regulated colonic mRNA expression levels of Reg3g and pro-inflammatory cytokines (IL-1β, IL-6, TNF-α) (P<0.05) while up-regulated that of anti-inflammatory factor IL-10 (P<0.000 1) when compared with the corresponding levels in AL mice. ABX treatment led no significant differences between the AL+ABX and TRF+ABX groups in term of DAI score, colon length, or histopathology. Obviously down-regulated Reg3g was observed in the TRF+ABX group than the AL+ABX group (P<0.05), whereas L-1β, IL-6, TNF-α and IL-10 showed no notable changes. 16S rRNA-seq revealed that TRF markedly reshaped gut microbiota composition, with increased Gram-positive bacterial abundance, reduced Gram-negative bacteria, with concomitant lower serum LPS level (P<0.001). RNA-seq also indicated significant suppression of NF-κB and other inflammation-related signaling pathways in the TRF group.

TRF attenuates DSS-induced colitis in mice by downr-egulating Reg3γ expression, reshaping gut microbiota, and reducing serum LPS level, and thereby suppressing NF-κB-mediated inflammatory signaling pathways.

To investigate the effects of fructose exposure on mitochondrial oxidative damage in M2-type macrophages and elucidate the regulatory role of protein phosphatase 2A (PP2A) in the process using its specific activator ABL127, an inhibitor of protein phosphatase methylesterase-1 (PPME-1).

① Immortalized mouse bone marrow-derived macrophages Ana-1 were subjected and grouped into M0 (conventional culture), M2 (treated with 20 ng/mL IL-4 for 24 h), and M2+Fru groups (IL-4 plus 0.04, 0.20, 1.00, or 5.00 mmol/L fructose). Cell viability was assessed with CCK-8 assay. Number of mitochondria, total and mitochondrial levels of reactive oxygen species (ROS), and mitochondrial membrane potential (ΔΨM) were measured using fluorescent probes. Total and demethylated PP2Ac protein levels were detected by Western blotting. ② Ana-1 cells were also divided into M0, M2, M2+Fru (20 ng/mL IL-4+5.00 mmol/L fructose, 24 h), and M2+Fru+ABL127 (20 ng/mL IL-4+5.00 mmol/L fructose+1.00 μmol/L ABL127, 24 h) to investigate PP2A-mediated mechanisms. Numbers of mitochondria and lysosomes, ROS level, and ΔΨM were detected via fluorescence assays. Expression of mitophagy-related proteins, PTEN induced putative kinase 1 (PINK1), P62, microtubule-associated protein light chain 3 (LC3), and voltage-dependent anion channel (VDAC) was evaluated by Western blotting, and the mRNA levels of M2 markers, found in inflammatory zone 1 (Fizz1), arginase-1 (Arg-1), and TGF-β were measured using RT-qPCR.

① Compared with the M2 group, fructose treatment at a concentration ranging from 0.04 to 5.00 mmol/L showed no effect on cell viability in M2 macrophages, but increased total ROS level in a dose-dependent manner (P<0.05). Fructose of 5.00 mmol/L resulted in significantly elevated mitochondrial ROS and mitochondrial quantity (P<0.05), reduced ΔΨM (P<0.05), up-regulated demethylated-PP2Ac (P<0.05), and no changed total-PP2Ac protein level. ② Compared with the M2 + Fru group, the addition of ABL127 led to decreased number of mitochondria but increased number of lysosomes (P<0.01), up-regulation of PINK1, LC3Ⅱ and VDAC proteins, down-regulation of P62 (P<0.05), reduced total and mitochondrial ROS levels, and enhanced ΔΨM (P<0.01). The mRNA expression of Fizz1, Arg-1, and TGF-β was notably decreased in the M2+Fru group than the M2 group (P<0.05), and the levels were rescued by ABL127 treatment (P<0.05).

Fructose induces PP2Ac demethylation and then mitochondrial oxidative damage in M2-type macrophages. PP2A activation promotes mitophagy and reverses fructose-induced damage.

To investigate the effect of Saikosaponin A (SSA) on the differentiation, apoptosis and function of mouse bone marrow derived macrophages (BMDM)-derived M1/M2 macrophages, and to explore its molecular mechanism.

BMDM was induced to differentiate into M1/M2 macrophages in vitro, and SSA was added at the same time: CCK-8 assay was used to detect the viability of BMDM and M1/M2 macrophages. The morphology of M1/M2 macrophages was observed by inverted fluorescence microscopy. Flow cytometry (FCM) and ELISA were used to detect the levels of surface markers and cytokines in M1/M2 macrophages. Real-time fluorescent quantitative PCR (qPCR) was used to detect the mRNA levels of IL-6, TNF-α and arginase-1 (Arg-1). FCM was used to detect the phagocytosis of peritoneal macrophages to fluorescent microsphere particles. Immunofluorescence (IF) assay and Western blotting were used to detect the molecular mechanism of SSA regulating M1/M2 macrophages.

No significant effect on viability of M1/M2 macrophages was observed at SSA concentration of 10.0 mg/L, and obvious inhibition was seen at a concentration of 15.0 mg/L (P<0.01). Treatment of 10.0 mg/L SSA induced obvious morphologic changes in M1/M2 macrophages, with M1 macrophages in irregular shape, a few having pseudopods, and some showing unclear boundaries; while some M2 macrophages presenting round or irregular (P<0.001) with unclear boundaries. SSA treatment also resulted in significantly decreased proportion of M1/M2 macrophages after BMDM differentiation (P<0.05), with reduced contents of IL-6 and TNF-α secreted by M1 macrophages and their mRNA levels (P<0.05), but increased secretion of Arg-1 and mRNA levels by M2 macrophages (P<0.05). SSA treatment also inhibited the phagocytosis ability of peritoneal macrophages to fluorescent microsphere particles (P<0.01) in a concentration-dependent manner. SSA decreased the phosphorylation of NF-kappaB (p-NF-κB) (P<0.01) and enhanced the phosphorylation of signal transducer and activator of transcription 6 (p-STAT6) in M2 macrophages (P<0.05).

SSA may affect the differentiation and function of M1/M2 macrophages by regulating NF-κB and STAT6 signaling pathways.

To explore the potential role and underlying mechanism of the functionally uncharacterized gene Gm49394 on regulating β-cell apoptosis under diabetic conditions.

The expression and translational activity of Gm49394 in pancreatic β-cell lines and non-β-cell lines were validated using RNA fluorescence in situ hybridization (RNA-FISH), quantitative real-time PCR (qPCR), Western blotting, and immunofluorescence (IF) assay. The β-cell lines (NIT-1/Min6) with Gm49394 overexpression or knockdown were constructed. The proliferation, apoptosis, mitochondrial function, as well as oxidative stress and endoplasmic reticulum stress markers in these β-cell lines under physiological homeostasis or pathological stress conditions, such as high glucose (30 mmol/L), inflammation (10 ng/mL IFN-γ alone or combined with 10 ng/mL IL-6), and hydrogen peroxide (100 μmol/L H₂O₂) were detected by flow cytometry and Western blotting.

RNA-FISH and qPCR indicated that Gm49394 was specifically expressed in pancreatic β-cell lines and up-regulated under high glucose or inflammatory stimulation. IF assay and Western blotting showed that Gm49394 had protein-coding activity. Flow cytometry and Western blotting identified that Gm49394 overexpression did not affect β-cell proliferation, but promoted β-cell apoptosis and increased reactive oxygen species (ROS) and mitochondrial superoxide (MitoSOX) levels in β cells under physiological homeostasis or pathological stress conditions (P<0.05). Under physiological conditions, Gm49394 knockdown failed to induce significant alterations on β-cell apoptosis, ROS, or MitoSOX levels. Under pathological stress conditions, Gm49394 knockdown significantly suppressed β-cell proliferation, apoptosis, as well as oxidative and endoplasmic reticulum stress (P<0.05).

Gm49394 may promote β-cell apoptosis via oxidative stress and endoplasmic reticulum stress.

To systematically identify the key genes regulating the exit from naïve pluripotency in embryonic stem cells (ESCs) in order to provide novel targets and theoretical insights into the mechanisms for pluripotency transition and early cell fate determination.

Nanog-green fluorescent protein (Nanog-GFP) reporter-labeled ESCs were infected with a genome-wide Brie knockout library, and further cultured under leukemia inhibitory factor/serum (LIF/S) conditions for 14 d. Flow cytometry was used to sort Nanog-GFP⁺(naïve-state) and Nanog-GFP^- (primed state) cell populations, followed by genomic DNA extraction and high-throughput sequencing. Model-based Analysis of Genome-wide CRISPR/Cas9 Knockout (MAGeCK) was applied to identify differential genes between GFP⁻/Input, GFP⁺/Input, and GFP⁺/GFP⁻ groups. Metascape and Gene Set Enrichment Analysis (GSEA) were conducted for functional enrichment analysis. Then the obtained candidate genes were employed to construct knockout models, and their roles were assessed through cell morphology observation, Nanog-positive rate detection, colony formation assays, and pluripotency gene expression analysis.

The GFP⁺/Input screening revealed 2 921 negatively regulated genes (mainly enriched in basic life processes, such as RNA metabolism and cell cycle) and 1 393 positively regulated genes (enriched in the processes of nervous system development, carbohydrate metabolism, and vascular system development). In the GFP⁻/Input screening, 2 765 negatively regulated genes (enriched in RNA metabolism, cell cycle, and other fundamental processes) and 1 303 positively regulated genes (enriched in neural development, cell survival, and endothelial migration) were identified. The GFP⁺/GFP⁻ comparison identified 1 001 negatively regulated genes [involved in stress response and inhibition of mitogen-activated protein kinase (MAPK) signaling] and 983 positively regulated genes [related to fibroblast growth factor/extracellular signal-regulated kinase (FGF/ERK) signaling pathway and glucose metabolism). These genes, were not only known pluripotency regulators (e.g., Nanog, Nr5a2, Klf2, Klf4) and exit-associated genes (e.g., Gata6, Grb2, Zeb1, Fgfr1), but also some novel candidates (e.g., Dmrt1, Rxra, Zbtb14 and Tmem41b). Functional validation showed that transient knockout of Dmrt1, Tmem41b, and Hic2 significantly increased the proportion of Nanog⁺ cells (P<0.01), suggesting their role in suppressing ground-state exit. ESCs with stable Dmrt1 knockout exhibited a more naïve-state phenotype, presenting compact, dome-shaped colonies, with increased ratio of undifferentiated colonies (P<0.01), up-regulation of ground-state markers (Nanog, Nr5a2, Dppa3, P<0.01), and down-regulation of primed-state markers (Fgf5, Lefty1, Dnmt3b, P<0.01). Rescue experiments for Dmrt1 expression reversed these above phenotypes.

A candidate gene set regulating exit from naïve pluripotency in ESC is screened out and identified with genome-wide CRISPR. Our findings implicate Dmrt1 plays a critical role in promoting the exit.

To investigate the impact of nucleophosmin 1-mutated (NPM1m) subtypes, variant allele frequency (VAF) and co-mutations on the survival outcomes of patients with acute myeloid leukemia (AML).

Clinical data, mutation status, and outcomes of 86 NPM1m-AML patients admitted in Department of Hematology, First Hospital of Lanzhou University between June 2017 and September 2024 were collected and retrospectively analyzed. Spearman correlation analysis, Kaplan-Meier curve analysis, Log-rank test, and Cox regression analysis was applied for correlation analysis, survival analysis, and factors affecting survival.

The overall survival (OS) was significantly shorter in the Rares subtype of NPM1m than the ABD subtype (median OS: 164 d vs 416 d, P=0.043). The VAF of NPM1mwas positively correlated with the initial peripheral blood white blood cell count and lactate dehydrogenase (LDH) level (P<0.05). OS was reduced when VAF was ≥0.37 (median OS: 164 d vs 730 d, P=0.003). The presence of myelodysplasia-related gene (MR) mutations was associated with a poorer prognosis when compared to the MR wild-type (median OS: 45 d vs 395 d, P<0.001). The triple mutation of FMS-like tyrosine kinase 3-internal tandem duplication (FLT3-ITD) and DNMT3A also indicated a worse prognosis than the non-triple mutation (median OS: 173 d vs 483 d, P=0.007). The presence of PTPN11 mutations was associated with improved OS (median OS: 395 d vs 240 d, P=0.027), while the patients with coexistence of N/KRAS mutations trended toward better prognosis than the wild-type patients (median OS: 662 d vs 189 d) though no statistical significance (P=0.070). Multivariate analysis revealed that LDH (HR=1.002, 95%CI: 1.000~1.003, P=0.005), NPM1m VAF (HR=2.415, 95%CI: 1.208~4.829, P=0.013), Rares subtype (HR=3.037, 95%CI: 1.134~8.136, P=0.027), and MR mutations (HR=5.283, 95%CI: 1.991~14.017, P<0.001) were independent risk factors associated with OS in the patients.

Molecular heterogeneity of NPM1m should be taken into account in prognostic stratification of NPM1m-AML. Factors including the Rares subtype, VAF≥0.37, coexistence of FLT3-ITD and DNMT3A mutations, and MR mutations are associated with poor prognosis of NPM1m-AML. The presence of PTPN11 mutations improves the prognosis, while the presence of N/KRAS mutations shows a trend toward better prognosis. LDH, NPM1m VAF, Rares subtype, and MR mutations are independent risk factors affecting OS of patients with NPM1m-AML.

To investigate the predictive value of a nomogram for central lymph node metastasis (CLNM) in patients with papillary thyroid microcarcinoma (PTMC) based on CT features combined with clinical factors.

A case-control study was conducted on 256 pathologically confirmed PTMC patients from 2 tertiary hospitals from January 2022 to November 2024. All participants underwent contrast-enhanced neck CT scanning within 2 weeks before surgery and received central lymph node dissection. The 201 patients from the First Affiliated Hospital of Army Medical University were randomized into a training set (n=140) and an internal validation set (n=61) in a 7∶3 ratio. The 55 patients from the Second Affiliated Hospital of Chongqing Medical University were all assigned into an external validation set. Their clinical data and CT features were collected. Univariate and multivariate logistic regression analyses were employed to identify independent predictive factors for CLNM, and then a nomogram model was constructed. Receiver operating characteristic (ROC) curve analysis (area under the curve, AUC), calibration curve, and decision curve analysis (DCA) were performed to evaluate the model performance, discrimination and clinical utility.

Multivariate logistic regression analysis identified 4 independent CLNM predictors (P<0.05), that is, male (OR=5.991, 95%CI: 2.209~18.350), tumor size ≥0.82 cm (OR=18.880, 95%CI: 1.803~229.500), capsular involvement (OR=9.805, 95%CI: 4.015~26.340), and CT-diagnosed lymph node positivity (OR=2.872; 95%CI: 1.176~7.232). The nomogram achieved an AUC value of 0.859 (95%CI: 0.796~0.922), 0.786 (95%CI: 0.671~0.901), and 0.783 (95%CI: 0.648~0.917) in the training and internal and external validation sets, respectively. Calibration curves demonstrated high consistency between predicted and observed probabilities (Hosmer-Lemeshow P>0.05). DCA confirmed net clinical benefits for CLNM before surgical treatment with a threshold probability range of 0.18~0.80.

Based on sex, tumor size, capsular involvement, and CT-diagnosed lymph node metastasis, our nomogram model effectively predicts CLNM risk in PTMC patients. It can be used as a quantitative tool for personalized surgical planning and shows high clinical applicability.