To investigate the mechanism by which peripubertal exposure to di-2-ethylhexyl phthalate （DEHP） induces reproductive injury in adult male mice through impairment of Sertoli cell function and the blood-testis barrier.

A total of 180 male C57BL/6J mice （3 weeks old） were randomly divided into corn oil group （control group），and 5，25，125，250 and 500 mg/kg DEHP exposure groups. All groups received daily oral gavage for 5 consecutive weeks. Ten mice from each group were sacrificed for first material collection when they were 9 weeks old. The remaining F0 generation mice were mated with unexposed adult female mice at a male-to-female ratio of 1∶2 at 14 weeks of age for a period of 2 weeks. The second material collection was completed at 16 weeks of age，with 20 mice taken from each group. A computer-assisted analysis system was used to detect sperm density and motility in 9-week-old and 16-week-old male mice. The average birth body weight of F1 generation pups，survival rate of pups at 4 d after birth per litter，number of pups per litter，and male-to-female sex ratio within the litter were recorded. ELISA was used to detect hormone levels in the serum. Sperm abnormalities were observed using eosin staining. HE staining was used to observe pathological changes in testicular tissue. Immunofluorescence assay was utilized to detect the protein expression of tight junction protein 1 （zonula occludens-1，ZO-1） and gap junction protein 43 （connexin43，Cx43） related to the blood-testis barrier in testicular tissue，as well as the protein expression of apoptosis-related molecule Cleaved-Caspase3. Immunohistochemistry was used to detect the expression of Sertoli cell marker SOX9 protein in testicular tissue. TUNEL was used to detect apoptosis in testicular tissue. Comparative toxicogenomics database （CTD） was used to screen for mono-2-ethylhexyl phthalate （MEHP） induced reproductive damage-associated genes and combined with GO/KEGG and Reactome enrichment analysis to predict possible signaling pathways. An in vitro exposure model was established by treating TM4 cells with MEHP at concentrations of 0，50，100，200 and 400 μmol/L，respectively. CCK-8 assay，flow cytometry and Western blotting were applied to detect the cell viability，apoptosis，and protein expression of apoptosis-related proteins B-cell lymphoma 2 （Bcl-2） and Bcl-2-associated X protein （Bax），and blood-testis barrier-related proteins ZO-1 and Cx43.

Compared with the control group，the pregnancy rate was decreased from 80.0% to 52.5%，and the total number of pups was decreased from 212 to 125 in the female mice mating with the male from the 500 mg/kg group. Compared with the control group，the sperm density and motility were significantly reduced，and the total rate of sperm abnormalities was obviously increased in the 9-week-old DEHP-exposed mice （P<0.05），and even in the 16-week-old mice，the sperm motility was still in a dose-dependent downward trend （P<0.05）. In the 25，125，250 and 500 mg/kg DEHP exposure groups，the thickness of the seminiferous epithelium was obviously decreased，with more cellular vacuolization and notable epithelial atrophy and degeneration. Cell apoptosis in the testicular tissue was enhanced，while the expression of ZO-1 and Cx43 were decreased in the 250 and 500 mg/kg exposure groups. The levels of androgen-binding protein，gonadotropin-releasing hormone，and testosterone in the serum were decreased in a dose-dependent manner （P<0.05），while those of luteinizing hormone and follicle-stimulating hormone showed a dose-dependent increase （P<0.05）. Bioinformatics analysis showed that 153 genes related to male system diseases were screened out by CTD for MEHP. GO enrichment，KEGG pathway，and Reactome enrichment analyses displayed that the apoptosis signaling pathway was significantly enriched. CCK-8 assay showed that TM4 cell viability was decreased in a dose-dependent manner after MEHP treatment when compared with the control group （P<0.05）. Western blotting indicated that MEHP treatment reduced the expression levels of Bcl-2，ZO-1 and Cx43 （P<0.05），and enhanced that of Bax in comparison with the control group （P<0.05）.

Peripubertal DEHP exposure may lead to reduced sperm quality and reproductive damage in adult male mice by disrupting the blood-testis barrier and activating Sertoli cell apoptosis.

To investigate the effects and potential mechanisms of polystyrene micro/nanoplastics (PS-MNPs) on testicular Sertoli cells.

Sixty male C57BL/6N mice (8 weeks old) were randomly divided into a control group (deionized water), a PS-NPs group [particle size of 20 nm, 2.5 mg/(kg·d)], and a PS-MPs group [particle size of 5 μm, 2.5 mg/(kg·d)], with 20 mice in each group. The corresponding agents were gavaged once daily for 6 months. HE staining was used to observe the histopathological and morphological changes in the testicular tissues. Immunohistochemistry of marker proteins was employed to evaluate the changes in the number of Sertoli cells. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were performed to identify functions and signaling pathways enriched in the testicular transcriptome. Mouse testicular Sertoli cell line TM4 was divided into a control group (deionized water), a 2.5NPs group (2.5 μg/mL), and a 2.5MPs group (2.5 μg/mL). All groups received continuous exposure through 130 cell passages. Cell viability and proliferative capacity were evaluated using CCK-8 assay and EdU incorporation, while cell migration was assessed using transwell and cell scratch assays. RT-qPCR and Western blotting were used to detect the changes in the expression of key molecules regulating mesenchymal phenotypic transformation (MPT) at mRNA and protein levels.

Pathological analysis revealed that, when compared to the control group, PS-NPs and PS-MPs treatment resulted in extended spaces between testicular seminiferous tubules, loosely arranged spermatogenic cells, and enhanced vacuolization. Immunohistochemical analysis of marker proteins indicated a decreasing trend in the number of testicular Sertoli cells in the PS-NPs and PS-MPs groups than the control group, with the PS-NPs group having statistical significance (P<0.01). GO and KEGG enrichment analyses revealed that PS-MNPs exposure-related altered genes were significantly enriched in cell adhesion signaling pathways (P<0.05). PS-MPs exposure significantly inhibited the growth and migration ability of TM4 cells (P<0.05), but PS-NPs exposure had no such effect on cell growth but notably enhanced cell migration ability. PS-NPs exposure inhibited the expression levels of E-cadherin and ZO-1 (P<0.01) and up-regulated the expression of N-cadherin and vimentin (P<0.01), and PS-MPs exposure led to significant up-regulation of vimentin (P<0.01) and down-regulation of E-cadherin, N-cadherin, and ZO-1 (P<0.05). Both PS-MPs and PS-NPs exposure up-regulated the mRNA levels of Snail2, Twist1, and Zeb2 (P<0.01).

Exposure of PS-MNPs leads to abnormal proliferation and migration of TM4 cells, induces decreases in cell-cell contacts among Sertoli cells and spermatogenic cells at all levels possibly through MPT, and thus results in testicular damage.

To investigate whether long-term low-dose exposure to polystyrene microplastics (PS-MPs) induces ferroptosis in testicular Sertoli cells and then leads to testicular injury.

Forty 8-week-old male C57BL/6 mice were randomly divided into a control group (deionized water group) and a PS-MPs group [2.5 mg/(kg·d)], with 20 mice in each group. Corresponding agents were gavaged once a day for 12 consecutive months. HE staining and Prussian blue staining were used to detect histopathological damage and accumulation of ferrous ions in the testes. Electron transmission microscopy was employed to observe the mitochondrial morphology of testicular Sertoli cells. Mouse Sertoli cell line TM4 was divided into a Con group (standard culture) and an MPs group (2.5 μg/mL PS-MPs). After both groups underwent continuous exposure and passed up to the 100th generation, morphological changes were observed under the microscope; cell viability was detected with CCK8 assay, and production of reactive oxygen species (ROS) and mitochondrial membrane potential (MMP) were detected with a ROS probe and a mitochondrial membrane potential probe (JC-1), respectively. Flow cytometry, ferrous ion (Fe²⁺) kit and Western blotting were applied to detect cell apoptosis, intracellular iron ion content, and protein levels of key molecules of ferroptosis in tissues and cells.

Long-term exposure to PS-MPs resulted in significantly reduced diameter and thickness of mouse varicocele (P<0.01), fewer testicular Sertoli cells (P<0.05), with characteristic ferroptosis alterations in the mitochondria, and increased accumulation of ferrous ions in testicular tissue. Exposure to PS-MPs down-regulated the key molecules of ferroptosis, glutathione peroxidase 4 （GPX4） and ferritin light chain （FTL） when compared with the control group (P<0.05). In the cell model, long-term PS-MPs exposure led to morphological changes and decreased cell viability (P<0.05), more production of ROS (P<0.01), and decrease in MMP (P<0.05) of TM4 cells. The exposure had no effect on cell apoptosis, but elevated the intracellular content of ferric ions (P<0.01), and down-regulated GPX4 and FTL protein levels (P<0.05).

Long-term low-dose exposure to PS-MPs induces mitochondrial damage and oxidative stress in testicular Sertoli cells, activates the ferroptosis pathway, and ultimately leads to testicular injury in mice.

To analyze the detrimental effects of exposure to environmental noise alone and combined with carbon monoxide (CO) on spermatogenesis in male rats, investigate the underlying mechanisms involved in such damage, and evaluate the protective role of pterostilbene (PTE) against these adverse effects.

Sixty male SD rats (6~8 weeks old, weighing 200±10 g) were randomly divided normal control group (standard housing), sham-exposure control group (restraint stress only), noise exposure (85 dB), CO exposure (460 mg/m³), combined exposure (simultaneous exposure), and PTE intervention (80 mg/kg pretreatment), with 10 animals in each group. The rats were exposed daily for 2 h via a nose-only inhalation exposure system within a multifactorial environmental simulation chamber for 60 consecutive days. Sperm count and viability were measured after exposure. Histopathological changes of testicular tissues were observed with HE staining. qRT-PCR was used to measure stage-specific mRNA levels in germ cells. Serum sex hormone levels and adenosine triphosphate (ATP) concentrations in testes and sperm were detected. Transmission electron microscopy (TEM) was applied to observe the ultrastructural damage in the spermatocytes. Additionally, transcriptome sequencing was performed on testicular tissue, followed by bioinformatics analysis.

Compared with the negative control group, the combined exposure group exhibited significant reductions in sperm viability and count (P<0.05), and developmental arrest of immature germ cells in the testicular tissue, with obviously less spermatogonia, spermatocytes, and round/elongated spermatids (P<0.01). Additionally, significantly reduced levels of reproductive-related hormones, such as gonadotropin-releasing hormone, follicle-stimulating hormone, luteinizing hormone and testosterone, and ATP levels in testes and sperm were observed in the mice after combined exposure (P<0.01), accompanied by mitochondrial rupture and cristae disruption in spermatocytes. Conversely, the PTE intervention group showed marked alleviation of these impairments, with parameters recovering almost to normal levels. Transcriptome sequencing identified biological processes related to reproductive development and ATP-dependent pathways as potential contributors to testicular injury induced by noise and CO exposure, with key genes including Nppa, Adm, Gnrh1, Ptafr, Atp13a5, Atp8b1, and LOC102555469.

Noise and CO exposure induce spermatogenic damage in rats, which may be related with energy metabolism and hormonal regulation, while PTE demonstrates significant protective effects against such reproductive impairments.

To investigate the transgenerational effects of paternal PM_2.5 exposure on offspring growth and development, and to preliminarily elucidate the role of sperm DNA methylation modifications in mediating these effects.

Eight-week-old male C57BL/6 mice were randomly divided into filtered air (FA), unfiltered air (UA), and concentrated PM_2.5 (CAP) groups, with 10 animals in each group. The exposure was conducted from November 2019 to April 2020, and then, these male mice were mated with unexposed females to generate F1 offspring, which were bred successively to produce F2 and F3 generations. All the offspring were living in PM_2.5-free environment. The birth body weight, birth number, and sex ratio of the offspring were recorded, body weight growth was monitored, and organ coefficients of the heart, liver, lung, and brain were calculated. Whole-genome methylation sequencing was performed on the sperm DNA of the CAP group, FA group, and their F1 generation offspring to screen for differentially methylated regions, and the genes and pathways associated with these regions were analyzed.

When compared with the F1~F3 offspring of the FA group, the CAP group had significantly reduced birth body weight in the F1 generation (P<0.05), no statistical differences were observed in the birth body weight in the F2 and F3 generations (P>0.05), or either in the sex ratio and birth number among the F1, F2 and F3 generations. Compared with the FA group offspring, the F1~F3 offspring of CAP group exhibited delayed body weight gain, especially in the males (P<0.05), the CAP-F1 male generation had obviously elevated liver organ coefficient (P<0.01), but no statistical changes were observed in the heart, lung, or brain coefficients among the F1~F3 generations. Between the FA group and the CAP group, 37 997 differentially methylated regions were detected, with a reduction of approximately 50% in the number of differentially methylated regions in the F1 generation. Differentially methylated genes in F0 and F1 sperm were potentially related to developmental processes, including imprinting genes (Gnas, Igf2) and metabolic genes (Ppard, Rps6kb1).

Paternal exposure to PM_2.5 leads to reduced birth weight and intergenerational growth retardation in offspring. Its impact on phenotypic effects is gradually weakened during intergenerational transmission. Changes in the methylation of development-related genes in sperm may be one of the mechanisms mediating this intergenerational effect.

To design and construct chimeric antigen receptor (CAR) T cells targeting Trop2, establish an in vitro cell exhaustion model through continuous antigen stimulation, and investigate their anti-tumor activity and exhaustion characteristics.

The second-generation CAR plasmid was constructed based on the single-chain variable fragment (scFv) sequence of Sacituzumab Govitecan targeting Trop2. The viral vector titer was determined by retroviral vector packaging and gradient dilution. Peripheral blood mononuclear cells (PBMCs) from healthy donors were isolated using Ficoll density gradient centrifugation, and CAR virus vectors were transduced into PBMCs activated with OKT-3/IL-2 to generate Trop2-targeted CAR T cells. CAR expression levels were assessed by flow cytometry using MYC tags. In vitro 3 tumor cell models were established, including human ovarian cancer cells (SKOV3), human breast cancer cells (MDA-MB-453), and human lung cancer cells (A549). The expression of the Trop2 antigen in these models was confirmed using flow cytometry. Additionally, luciferase assay was employed to evaluate the cytotoxic efficiency of Trop2-targeted therapy at various effector-to-target ratios. An in vitro CAR-T exhaustion model was developed, and the long-term killing ability of CAR-T cells was dynamically monitored using the Incucyte live-cell imaging system. The PD-1/TIM-3 phenotype of CAR-T cells was analyzed by flow cytometry, and cytokine secretion levels were quantified using the cytometric bead array (CBA). Transcriptomic sequencing and RT-qPCR were employed to validate the differentially expressed genes associated with exhaustion.

The second-generation CAR T cells targeting Trop2 were successfully constructed. Compared to the P-T group, in vitro experiments demonstrated that these CAR T cells exhibited antigen-specific and dose-dependent cytotoxic effects against tumor cells with high Trop2 expression, such as MDA-MB-453 and SKOV3. A CAR-T cell exhaustion model established through repeated tumor antigen stimulation in vitro revealed that, compared to the initial state, the exhausted Trop2 CAR-T cells exhibited significantly reduced tumor-killing capacity while P-T cells showed almost no killing effect, the expression of inhibitory receptors (PD-1 and TIM-3) was up-regulated on the surface of exhausted CAR-T cells, and the secretion of effector cytokines was diminished. Transcriptomic analysis identified multiple differentially expressed genes in the exhausted CAR-T cells. Pathways related to immune response and T cell receptor signaling were down-regulated, while apoptosis-related pathways were activated. RT-qPCR further confirmed abnormal expression of immunoregulatory genes, including IL3, IL5, and IL13 (P<0.05).

During continuous in vitro tumor antigen stimulation, the second-generation CAR-T cells targeting the Trop2 antigen demonstrate declined anti-tumor activity, weakened effector function and up-regulated expression of exhaustion-related molecules.

To develop novel metformin carbon dots (MCDs) with superior performance to enhance the osteogenic differentiation potential of human periodontal ligament stem cells (hPDLSCs) under inflammatory conditions.

Three types of MCDs-MCDsCA, MCDsAA, and MCDsCS were synthesized via a one-step hydrothermal method using metformin (Met) in combination with citric acid (CA), ascorbic acid (AA), and carboxymethyl chitosan (CS), respectively. Their physicochemical properties were characterized by transmission electron microscopy (TEM), Fourier-transform infrared spectroscopy (FTIR), and UV-visible spectrophotometry. The biocompatibility and cellular uptake of the MCDs were assessed using CCK-8 assay and confocal laser scanning microscopy. Alkaline phosphatase (ALP) staining and ALP activity assay were conducted to identify the most effective MCDs type in promoting osteogenic differentiation of hPDLSCs under inflammatory conditions. Subsequently, ALP and Alizarin Red S (ARS) staining, qRT-PCR, and Western blotting were performed to assess the effects of the optimal concentration of MCDsCS and its raw material (Met+CS) on osteogenic differentiation under inflammatory conditions.

All 3 types of nanoscale MCDs exhibited favorable physicochemical characteristics and biocompatibility, and could be effectively internalized by hPDLSCs. Both MCDsCA and MCDsAA significantly promoted hPDLSCs proliferation at lower concentrations (0~0.50 mg/mL), while MCDsCS exhibited excellent cytocompatibility over a broader range (0~2.00 mg/mL). Under inflammatory conditions, MCDsCA and MCDsAA showed no significant effect on ALP expression in hPDLSCs, whereas MCDsCS significantly enhanced ALP expression in a dose-dependent manner, with the optimal effect observed at 0.10 mg/mL (P<0.05). Compared to Met+CS, 0.10 mg/mL MCDsCS significantly enhanced ALP expression and calcium nodule formation under inflammatory conditions. It also upregulated the expression of osteogenesis-related genes (ALP, RUNX2, COL-1, OCN) and proteins (ALP, RUNX2, OCN) (P<0.01), while reducing the levels of inflammatory cytokines IL-1β and IL-6 (P<0.01).

A novel MCDsCS with sound biocompatibility is successfully developed, and can effectively promote osteogenic differentiation of hPDLSCs and exerts anti-inflammatory effects under inflammatory conditions, indicating its potential as a nanotherapeutic agent for periodontitis-associated bone regeneration.

To investigate whether remimazolam attenuates intestinal ischemia-reperfusion (I/R) injury in mice by regulating ferroptosis through connexin-43 (CX43).

Molecular docking was applied to predict the binding affinity of remimazolam to CX43. A total of 72 SPF-grade adult male C57BL/6J mice (6~8 weeks old, weighing 20~25 g) were subjected. Thirty of them were randomly divided into sham operation group (Sham group), I/R group 1, and I/R+10, 20 and 40 mg/kg remimazolam groups (RM10, RM20 and RM40 groups), with 6 mice in each group. Another 30 mice were randomly assigned into 5 groups (n=6), I/R group 2, erastin group (E group), I/R+40 mg/kg remimazolam group 2 (RM40 group 2), I/R+Fer-1 group (Fer-1 group), and erastin +40 mg/kg remimazolam group (ERM group). The left 12 mice were randomly and equally grouped into I/R+RM+oe-NC group (oe-NC group) and I/R+RM+oe-CX43 group (oe-CX43 group). The Fer-1 group was given an intraperitoneal injection of 5 mg/kg Fer-1 in 1 h prior to reperfusion, the E group was given 10 mg/kg erastin intraperitoneally 1 d before modeling, and all the remimazolam groups, the oe-NC group and the oe-CX43 group were injected intravenously with corresponding doses of remimazolam 30 min pre-modeling, while the oe-NC and oe-CX43 groups were injected with empty vector virus and overexpression of CX43 vector virus, respectively, 48 h before the administration of remimazolam. A mouse intestinal I/R injury model was constructed by clamping the superior mesenteric artery for 45 min and reperfusion for 30 min. The small intestine tissues were harvested and observed for pathological changes, and the intestinal mucosal damage was assessed with Chiu’s score. The contents of Fe²⁺, total iron, malondialdehyde (MDA), glutathione (GSH), and superoxide dismutase (SOD) were detected by colorimetric assay; the production of reactive oxygen species (ROS) was determined by DHE probe; the expression of ferroptosis-related genes was determined by RT-qPCR; and the expression levels of CX43, GPX4, and SLC7A11 were detected by Western blotting.

Molecular docking indicated that remimazolam had a binding energy of -6.699 kcal/mol with CX43 protein, suggesting good binding affinity between them. Compared with the Sham group, the I/R group 1 showed increases in Chiu’s scores and CX43 expression (P<0.05), along with pathological damage to intestinal tissues, and elevated contents of Fe²⁺, total iron, ROS and MDA (P<0.05), and down-regulated GPX4 and SLC7A11 (P<0.05). Compared with the I/R group 1, Chiu’s score was reduced in the RM40 group, and CX43 was significantly down-regulated (P<0.05), contents of Fe²⁺, total iron, ROS, and MDA were decreased (P<0.05), and expression levels of GPX4 and SLC7A11 were enhanced (P<0.05), and severity of intestinal histological damage was attenuated in both the RM40 and Fer-1 groups. Compared with the E group, the ERM group had the decreases in CX43 expression level (P<0.05), Fe²⁺, total iron, ROS, and MDA contents (P<0.05), and increases in GPX4 and SLC7A11 expression levels (P<0.05), with the improvement in intestinal tissue. Compared with the oe-NC group, overexpression of CX43 resulted in the increased CX43 expression, elevated contents of Fe²⁺, total iron, ROS and MDA (P<0.05) and decreased expression of GPX4 and SLC7A11 (P<0.05), leading to the exacerbated injury in intestinal tissue.

Remimazolam attenuates intestinal I/R injury by inhibiting ferroptosis through down-regulating CX43 expression.

To prepare humanized monoclonal antibodies (Mabs) targeting Acinetobacter baumannii (Ab) based on single memory B cell sequencing technology, construct the immune repertoire of the core protein of Ab, hemolysin-coregulated protein (Hcp), and express its Mabs with binding activity.

E. coli BL21 harboring the recombinant plasmid pGEX-6p-1-Hcp was constructed. Hcp protein was obtained using protein expression and affinity chromatography. Female SPF BALB/c mice (6~8 weeks old, weighing 18~20 g) were immunized intramuscularly with antigen Hcp to generate specific memory B cells. Single antigen-specific memory B cells were sorted using flow cytometry. The immune repertoire of Hcp was constructed using single-cell sequencing technology, and bioinformatics analysis was performed on the sequencing results. Mabs were obtained using antibody humanization techniques. The in vitro binding activity of the antibodies was detected by ELISA.

The target protein Hcp with a purity >95% was obtained after expression and purification. The immune repertoire of Hcp was successfully constructed, and the results of BCR clonotype identification and analysis, CDR3 region characteristic analysis, and V-J gene pairing characteristic analysis were achieved. Antibody humanization got 7 Mabs, that is, IgG1-1, IgG1-2, IgG2-1, IgG2-2, IgG3-1, IgG4-1 and IgG4-2. ELISA results showed IgG1-1, IgG3-1, IgG4-1, and IgG4-2 had an antibody binding titer of 1∶1 280, IgG2-2 of 1∶10 240, IgG2-1 of 1∶5 120, and IgG1-2 of 1∶160.

Single-cell sequencing technology enables rapid, accurate, and efficient construction of an Hcp protein immune repertoire containing extensive antibody information. Utilizing this immune repertoire allows for the expression of Mabs with binding activity.

To evaluate the predictive value of triglyceride-glucose body mass index (TyG-BMI), atherogenic index of plasma (AIP), and postprandial glucose excursion (PPGE) for sarcopenia in patients with type 2 diabetes mellitus (T2DM).

An observational cohort trial was conducted on 590 hospitalized T2DM patients (322 males and 268 females) admitted to Department of Endocrinology of the First Affiliated Hospital of Chongqing Medical University between January 2023 and December 2024. General demographic data and key metabolic indicators, including fasting plasma glucose (FPG), postprandial 2-hour plasma glucose (PPG), triglyceride (TG) were collected, and then TyG-BMI, AIP, and PPGE were calculated. Based on the Asian Working Group for Sarcopenia (AWGS) 2019 criteria, they were divided into a sarcopenia group (n=140) and a non-sarcopenia group (n=450). After 1∶1 propensity score matching (PSM), 102 patients with sarcopenia were matched with 102 without. Differences in metabolic indicators FPG, PPG, TyG-BMI, AIP and PPGE were compared between the 2 groups. Spearman correlation analysis was conducted to assess correlations of these indicators with sarcopenia. Conditional logistic regression analysis was performed to identify independent risk factors, and receiver operating characteristic (ROC) curves were plotted to assess predictive performance. An external validation cohort consisting of 192 T2DM patients from Department of Endocrinology of Jiangbei Branch of First Affiliated Hospital of Army Medical University between January 2023 and December 2024 was included to validate the prediction model.

After PSM, the baseline data of the 2 groups was balanced (P>0.05). The sarcopenia group showed higher levels of PPG, AIP, and PPGE, but lower TyG-BMI value than the non-sarcopenia group (all P=0.001). Spearman correlation analysis revealed that the TyG-BMI was negatively correlated (r=-0.404, P=0.001), whereas AIP and PPGE were positively correlated with concomitant sarcopenia (r=0.280,P=0.001;r=0.372, P=0.001) in the T2DM patients. Conditional logistic regression analysis identified AIP (OR=14.367) and PPGE (OR=1.245) as independent risk factors, while TyG-BMI (OR=0.966) as independent protective factors. ROC curve analysis revealed that the area under the curve (AUC) of the combined model of TyG-BMI, AIP and PPGE in predicting sarcopenia was 0.918, and the AUC value was 0.954 in external validation, with good sensitivity and specificity.

TyG-BMI, AIP, and PPGE are important metabolic predictors in T2DM patients with sarcopenia. Their combination has good screening value for sarcopenia, and can be applied in external prediction for T2DM patients.

To compare the effects of main artery clamping (MAC) and selective artery clamping (SAC) strategies on postoperative renal function in patients with chronic renal insufficiency undergoing robot-assisted partial nephrectomy.

A retrospective cohort study was conducted on 231 patients with preoperative chronic renal insufficiency [eGFR <90 mL/(min·1.73 m²) with renal injury markers or eGFR <60 mL/(min·1.73 m²)] who underwent robot-assisted partial nephrectomy in the Department of Urology of the First Affiliated Hospital of Army Medical University from February 2018 to February 2024. According to intraoperative renal artery clamping strategy, they were divided into a MAC group (n=129) and a SAC group (n=102). Preoperatively, individualized renal artery clamping strategies were developed using a machine learning-based multimodal holographic 3-D reconstruction technique. Serum creatinine (Scr) level was measured at 3 d and 3 months after surgery, and estimated glomerular filtration rate (eGFR) was calculated using the chronic kidney disease epidemiology collaboration equation (CKD-EPI) formula. Renal dynamic imaging with 99mTc-DTPA or 99mTc-MAG3 was used to assess the GFR of the affected kidney.

At 3 d after surgery, the decrease in GFR of the affected kidney was significantly lower [(8.3±7.7) vs (16.0±10.2) mL/(min·1.73 m²), 95%CI: -10.2~-5.2, P<0.001] in the SAC group than the MAC group. Scr increment analysis showed that the SAC group exhibited notably lower Scr increase [8.2 (2.5, 18.7) vs 15.5 (5.8, 28.3) μmol/L, 95%CI: -12.3~-1.8, P=0.027], and milder eGFR decline [3.0 (0.5, 7.8) vs 7.5 (2.0, 14.3) mL/(min·1.73 m²), 95%CI: -6.2~-0.8, P=0.015]. And, in 3 months after surgery, the SAC group had lower Scr level [(89.2±23.1) vs (95.3±22.1) μmol/L, 95%CI: -11.9~-0.3, P=0.042], and higher GFR of the affected kidney [(33.5±10.5) vs (26.1±10.9) mL/(min·1.73 m²), 95%CI: 4.6~10.2, P<0.001].

For patients with chronic renal insufficiency undergoing robot-assisted partial nephrectomy, SAC strategy is superior to MAC strategy in protecting postoperative renal function without increasing surgical risk.

To investigate the protective effects of remimazolam (Remi), a novel benzodiazepine sedative, on intestinal barrier function in septic mice.

A mouse model of sepsis was established using cecal ligation and puncture (CLP). A total of 96 SPF-grade adult male C57BL/6 mice were randomized into sham operation (Sham), sepsis (Sepsis), and sepsis with Remi intervention (Sepsis+Remi) groups. Survival rate and survival time were recorded within 72 h after modeling. Intestinal pathological alterations, barrier functional indicators, ZO-1 expression, and macrophage polarization status were observed and detected to evaluate the effects of Remi. Lipopolysaccharide (LPS) was used to treat RAW264.7 cells for 24 h to simulate in vitro sepsis model. The cells were divided into control (Control), LPS, and LPS+Remi groups. Immunofluorescence staining was performed to assess macrophage phenotype, mitochondrial morphology, and mitochondrial reactive oxygen species (MtROS), and Western blotting was applied to detect the protein expression of iNOS and CD206.

Compared with the sepsis group, Remi intervention significantly improved the survival rate of septic mice from 12.50% to 68.75% and markedly prolonged survival duration (P<0.05). Histopathological analysis demonstrated partial restoration of intestinal villus architecture, accompanied with attenuated interstitial edema and reduced inflammatory cell infiltration after Remi intervention. Furthermore, the intervention group demonstrated significant improvement in functional indicators. Both in vivo and in vitro experiments demonstrated elevated iNOS and decreased CD206 expression in the septic mice and LPS-stimulated macrophages（P<0.05）, which were partially reversed after Remi intervention. Furthermore, LPS-stimulated macrophages exhibited fragmented mitochondria and elevated MtROS level, whereas Remi intervention ameliorated these conditions（P<0.05）.

Remi protects intestinal barrier function in septic mice by mitigating mitochondrial dynamics imbalance-induced oxidative damage and ameliorating inflammatory macrophage activation.

To investigate the effects of long-term radiofrequency (RF) radiation at 2.65 GHz on behavior, inflammatory response, and intestinal microecology in mice in order to provide a basis for the safety assessment of long-term RF exposure.

One hundred and eight male C57BL/6N mice (17~21 g, 6~8 weeks old) were randomly assigned to a control group (Con) and a RF exposure group. The mice of the RF exposure group were subjected to whole-body uniform exposure to 2.65 GHz RF radiation in an electromagnetic reverberation chamber for 3 h/day for 28 consecutive days. RF field distribution and changes in core body temperature were monitored using an electromagnetic radiation analyzer and a fiber-optic temperature probe, respectively. Cognitive function was assessed using the Y-maze and novel object recognition (NOR) test. Anxiety-like behaviors were evaluated through open field test (OFT) and elevated plus maze (EPM), while depressive-like behaviors were examined with sucrose preference test (SPT) and tail suspension test (TST). HE staining was used to observe the histopathological changes in mouse tissues. Radioimmunoassay (RIA) was employed to detect the expression of pro-inflammatory cytokines, TNF-α and IL-1β, as well as anti-inflammatory cytokines, IL-4 and IL-10 in the serum, brain, jejunum, and spleen samples. Additionally, metagenomic sequencing was performed to assess alterations in the gut microbiota composition.

Long-term RF radiation led to a maximal increase of 0.59 ℃ in the core body temperature, but had no significant effects on cognitive function, anxiety-like behaviors, or depressive-like behaviors, or apparent damage of the hippocampal or jejunal tissues in the exposed mice. However, RF exposure significantly up-regulated the expression of the pro-inflammatory cytokine TNF-α in serum (P<0.05), and did not significantly alter the concentrations of other cytokines (IL-1β，IL-4，IL-10), caused significant decrease in α-diversity of the intestinal microbiota (P<0.01), with reduced relative abundances of Ligilactobacillus murinus and Acetatifactor muris (P<0.05), while elevated abundances of Lachnospiraceae bacterium (P<0.01).

Long-term exposure to 2.65 GHz RF radiation induces systemic inflammatory responses and disrupts gut microbiota homeostasis in mice.