Background: Bushen Zhuanggu Formula (BZF), derived from the classic Yougui Pills, has shown favorable clinical efficacy in treating advanced nontraumatic osteonecrosis of the femoral head (NONFH), particularly by promoting bone repair. However, its underlying mechanisms remain unclear. Objective: This study aimed to explore the mechanisms by which BZF promotes bone repair in advanced NONFH. Materials and methods: A total of 518 potential BZF targets were identified from the ETCM v2.0 database. Transcriptomic profiling of clinical cohorts revealed 485 differentially expressed genes in advanced NONFH patients compared to healthy controls. A drug target-disease gene interaction network was constructed to identify candidate BZF targets involved in NONFH pathogenesis. In vivo experiments were conducted to validate the effects of BZF in a rat model of advanced NONFH. Results: Network analysis identified key pathways associated with blood circulation obstruction, immune-inflammatory imbalance, and abnormal bone metabolism. Protein kinase C alpha (PKCA), Ras proto-oncogene (RAS), mitogen-activated protein kinase 3(ERK), ETS proto-oncogene 1 (ETS1), and receptor activator of nuclear factor-κB ligand (RANKL) formed a signaling axis implicated in NONFH pathogenesis. BZF treatment alleviated joint inflammation, preserved trabecular bone morphology, reduced bone loss, and promoted bone repair. Mechanistically, BZF significantly downregulated the expression of PKCA, RAS, ERK, ETS1, and RANKL, improved blood circulation, and inhibited osteoclast activation while promoting osteoblast activation. Conclusion: BZF may promote bone repair in advanced NONFH by enhancing blood circulation and modulating the PKC-RAS-ERK-ETS1-RANKL signaling axis, thereby reversing dysregulated bone metabolism.

Objective:To evaluate whether the mechanism by which mesenchymal stem cell-derived exosomes (MSC-exo) mitigated endotoxin-induced lung injury was related to the heme oxygenase-1 (HO-1)/mitochondrial signaling pathway in alveolar macrophages of mice.Methods:In vivo experiment Eighteen C57BL/6 wild-type (WT) mice were divided into 3 groups ( n=6 each) using a random number table method: control group (C group), lipopolysaccharide (LPS) group (L group) and LPS + MSC-exo group (LM group). Six HO-1 conditional knockout mice (HO-1 -/-) were selected and served as HO-1 -/- + MSC-exo + LPS group (HML group). The model of endotoxin-induced lung injury was prepared by injection of LPS 15 mg/kg. MSC-exo (2×10 11 particles) was intravenously injected at 1 h before injection of LPS in LM group. MSC-exo (2×10 11 particles) was intravenously injected and 1 h later LPS was injected in HML group. The expression of HO-1 in macrophages was detected using immunofluorescence, lung injury was assessed following hematoxylin-eosin staining, the wet/dry weight ratio (W/D ratio) was determined, and the mitochondrial morphology was observed with a transmission electron microscope. Cell experiment Alveolar macrophages (MH-S) were divided into 4 groups ( n=20 each) using a random number table method: control group (C group), LPS+ phosphate buffer solution group (LP group), LPS+ MSC-exo group (LM group), and LPS+ MSC-exo+ HO-1 small-interfering RNA group (LMS group). Cells were incubated for 12 h with LPS 10 μg/ml in LP, LM and LMS groups. In addition, LM group was incubated with MSC-exo 100 μg/ml, LP group was incubated with the equal volume of phosphate buffer solution, and the alveolar macrophages were transfected with HO-1 small interfering RNA and incubated with MSC-exo 100 μg/ml in LMS group at the same time. The concentrations of interleukin-1beta (IL-1β) and tumor necrosis factor-alpha (TNF-α) in supernatant were measured by enzyme-linked immunosorbent assay, HO-1 expression was detected by Western blot, the mitochondrial membrane potential was measured using JC-1 staining, and the expression of reactive oxygen species (ROS) was detected by fluorescence. Results:In vivo experiment Compared to C group, the lung injury score and W/D ratio were significantly increased ( P<0.05), the fluorescence signal of HO-1 in macrophages was enhanced, and the damage to mitochondria was aggravated in L group. Compared to L group, the lung injury score and W/D ratio were significantly decreased ( P<0.05), the fluorescence signal of HO-1 in macrophages was enhanced, and the damage to mitochondria was reduced in LM group. Compared to LM group, the lung injury score and W/D ratio were significantly increased ( P<0.05), macrophages had no HO-1 fluorescence signal, and the damage to mitochondria was aggravated in HML group. Cell experiment Compared to C group, the concentrations of IL-1β and TNF-α in supernatant were significantly increased, the expression of HO-1 was up-regulated ( P<0.05), and the mitochondria predominantly exhibited green JC-1 fluorescence, accompanied by an enhanced ROS fluorescence signal in LP group. Compared to LP group, the concentrations of IL-1β and TNF-α in supernatant were significantly decreased, the expression of HO-1 was up-regulated ( P<0.05), and the mitochondria predominantly exhibited red JC-1 fluorescence, accompanied by a weakened ROS fluorescence signal in LM group. Compared to LM group, the concentrations of IL-1β and TNF-α in supernatant were significantly increased, the expression of HO-1 was down-regulated ( P<0.05), and the mitochondria predominantly exhibited green JC-1 fluorescence, accompanied by an enhanced ROS fluorescence signal in LMS group. Conclusions:The mechanism by which MSC-exo attenuates endotoxin-induced lung injury may be related to up-regulation of HO-1 expression in alveolar macrophages and reduction of mitochondrial damage in mice.

Background:Bushen Zhuanggu Formula(BZF),derived from the classic Yougui Pills,has shown favorable clinical efficacy in treating advanced nontraumatic osteonecrosis of the femoral head(NONFH),particularly by promoting bone repair.However,its underlying mechanisms remain unclear.Objective:This study aimed to explore the mechanisms by which BZF promotes bone repair in advanced NONFH.Materials and methods:A total of 518 potential BZF targets were identified from the ETCM v2.0 database.Transcriptomic profiling of clinical cohorts revealed 485 differentially expressed genes in advanced NONFH patients compared to healthy controls.A drug target-disease gene interaction network was constructed to identify candidate BZF targets involved in NONFH pathogenesis.In vivo experiments were conducted to validate the effects of BZF in a rat model of advanced NONFH.Results:Network analysis identified key pathways associated with blood circulation obstruction,immune-inflammatory imbalance,and abnormal bone metabolism.Protein kinase C alpha(PKCA),Ras proto-oncogene(RAS),mitogen-activated protein kinase 3(ERK),ETS proto-oncogene 1(ETS1),and receptor activator of nuclear factor-κB ligand(RANKL)formed a signaling axis implicated in NONFH pathogenesis.BZF treatment alleviated joint inflammation,preserved trabecular bone morphology,reduced bone loss,and promoted bone repair.Mechanistically,BZF significantly downregulated the expression of PKCA,RAS,ERK,ETS1,and RANKL,improved blood circulation,and inhibited osteoclast activation while promoting osteoblast activation.Conclusion:BZF may promote bone repair in advanced NONFH by enhancing blood circulation and modulating the PKC-RAS-ERK-ETS1-RANKL signaling axis,thereby reversing dysregulated bone metabolism.

Objective:To understand the contract renewal willingness and its influencing factors among rural order-oriented medical students in Shaanxi Province. M ethods This study employed an explanatory sequential mixed methods design to examine contract renewal patterns among rural order-oriented medical graduates. From February to July 2024, a questionnaire survey was conducted among rural order-oriented medical students who graduated from 2015 to 2023 in Shaanxi Province. The participants were first stratified into three strata based on their year of graduation and stage of service, and one-third of each stratum was randomly selected as the research subjects. Univariate and multivariate analysis methods were used to explore the influencing factors of their willingness to renew their service in rural areas. Secondly, qualitative research methods were employed to conduct thematic interviews with 36 targeted medical students on the influencing factors of their willingness to renew their service. Results:A total of 513 valid questionnaires were collected during the quantitative research phase, including 224 males and 289 females. Of these, 14 were from the 2015-2017 cohort, 247 from the 2018-2020 cohort, and 252 from the 2021-2023 cohort. The results showed that only 30.4%(156/513) of the orientation medical students were willing to practice in primary care after the period of service. Univariate analysis showed that there were six factors related to the willingness to renew the contract, the consistency of the source and implementation of the contract, the completion of the standardized training of residents, the satisfaction with primary work, professional identity, the ability of primary diagnosis and treatment, and the training system and the suitability of primary work. Multivariate analysis showed that the willingness to renew the contract was significantly higher in the students who had the same place of origin and the place of performance( OR=1.7, 95% CI: 1.1-2.6, P=0.022). The willingness to renew the contract was significantly higher among students who participated in the standardized residency training than those who completed the training( OR=2.0, 95% CI: 1.3-3.0, P=0.003), and students with a better fit between the training system and working in primary care were more likely to renew their contract( OR=4.1, 95% CI: 2.8-6.0, P<0.001). Four themes were extracted from the interview: subjective factors, objective environment, policy factors and other factors. Conclusions:The study shows that improving the consistency of the source of students and the implementation of the contract, strengthening the standardized training of residents, and optimizing the adaptation of the training system to the primary work are the key measures to improve the willingness of directional medical students to renew their contract at the primary level.

OBJECTIVES: As early as the Apollo 11 mission, astronauts experienced ocular, skin, and upper airway irritation after lunar dust (LD) was brought into the return cabin, drawing attention to its potential biological toxicity. However, the biological effects of LD exposure through the digestive system remain poorly understood. This study aimed to evaluate the impact of digestive exposure to lunar dust simulant (LDS) on gut microbiota and on the intestine, liver, kidney, lung, and bone in mice. METHODS: Eight-week-old female C57BL/6J mice were used. LDS was used as a substitute for lunar dust, and Shaanxi loess was used as Earth dust (ED). Mice were randomly divided into a phosphate buffered saline (PBS) group, an ED group (500 mg/kg), and a LDS group (500 mg/kg), with assessments at days 7, 14, and 28. Mice were gavaged once every 3 days, with body weight recorded before each gavage. At sacrifice, fecal samples were analyzed by 16S ribosomal RNA (rRNA) sequencing; inflammatory cytokine expression [interleukin (IL)-1β, IL-6, and tumor necrosis factor alpha (TNF-α)] in intestinal, liver, and lung tissues was measured by real-time reverse transcription PCR (real-time RT-PCR); hematoxylin and eosin (HE) staining was performed on lung, liver, and intestinal tissues; Periodic acid-Schiff (PAS) staining was used to assess the integrity of the intestinal mucus barrier, and immunohistochemical staining was performed to evaluate the expression of mucin-2 (MUC2). Serum biochemical tests assessed hepatic and renal function. Femoral bone mass was analyzed by micro-computed tomography (micro-CT); osteoblasts and osteoclasts were assessed by osteocalcin (OCN) and tartrate-resistant acid phosphatase (TRAP) staining. Bone marrow immune cell subsets were analyzed by flow cytometry. RESULTS: At day 10, weight gain was slowed in ED and LDS groups. At days 22 and 28, body weight in both ED and LDS groups was significantly lower than controls (both P<0.05). LDS exposure increased microbial species richness and diversity at day 7. Compared with the PBS and ED groups, mice in the LDS group showed increased relative abundance of Deferribacterota, Desulfobacterota, and Campylobacterota, and decreased Firmicutes, with increased Helicobacter typhlonius and reduced Lactobacillus johnsonii and Lactobacillusmurinus. HE and PAS staining of the colon showed that mucosal structural disruption and goblet cell loss were more severe in the LDS group. In addition, immunohistochemistry revealed a significant downregulation of MUC2 expression in this group (P<0.05). No obvious pathological alterations were observed in liver HE staining among the 3 groups, and none of the groups exhibited notable hepatic or renal dysfunction. HE staining of the lungs in the ED and LDS groups showed increased perivascular inflammatory cell infiltration (both P<0.05). CONCLUSIONS LDS exposure via the digestive route induces gut dysbiosis, intestinal barrier disruption, pulmonary inflammation, bone loss, and bone marrow immune imbalance. These findings indicate that LD exposure poses potential health risks during future lunar missions. Targeted restoration of beneficial gut microbiota may represent a promising strategy to mitigate LD-related health hazards.
Animals ; Dust ; Mice ; Mice, Inbred C57BL ; Dysbiosis/etiology* ; Female ; Gastrointestinal Microbiome/drug effects* ; Moon ; Liver/metabolism* ; Digestive System/microbiology* ; Lung/metabolism* ; Kidney

OBJECTIVES: Due to prolonged exposure to cosmic radiation and meteorite impacts, lunar surface dust forms nanoscale angular particles with strong electrostatic adsorption properties. These dust particles pose potential inhalation risks, yet their pulmonary toxicological mechanisms remain unclear. Given the need for dust exposure protection in future lunar base construction and resource development, this study established an acute exposure model using lunar soil simulant (LSS) and used Earth soil (ES; Loess from Shaanxi, China) as a comparison to investigate lung injury mechanisms. METHODS: C57BL/6 mice were randomly assigned to 3 groups: Phosphate buffered saline (PBS), LSS, and ES, with 5 to 7 mice per group. Mice in the LSS and ES groups received a single intratracheal instillation to induce acute inhalation exposure. Body weight was monitored for 28 days. Mice were euthanized at days 3, 7, 14, and 28 post-exposure, and peripheral blood, bronchoalveolar lavage fluid (BALF), and lung tissues were collected. Immune cell subsets in BALF were analyzed using flow cytometry. Hematoxylin-eosin (HE) staining assessed lung structure and inflammation; periodic acid-Schiff (PAS) staining evaluated airway mucus secretion; Masson staining examined collagen deposition. Real-time reverse transcription PCR (real-time RT-PCR) was used to measure the mRNA expression of inflammatory cytokines (IL-1β, IL-6, and TNF-α) and epithelial barrier genes (Occludin, Cadherin-1, and Zo-1). Lung tissues at day 7 were subjected to transcriptomic sequencing, followed by immune infiltration and pathway enrichment analyses to determine immunoregulatory mechanisms. RESULTS: Body weight in the ES group progressively declined after day 18 (all P<0.05), while the LSS group showed no significant changes compared with the control group. HE staining showed both LSS and ES induced inflammatory cell infiltration around airways and vasculature, which persisted for 28 days but gradually lessened over time. PAS staining revealed marked mucus hypersecretion in the LSS group at day 3, followed by gradual recovery; no significant mucus changes were observed in the ES group. Masson staining indicated no obvious pulmonary fibrosis in either group within 28 days. Real-time RT-PCR demonstrated significant upregulation of IL-1β and TNF-α in both LSS and ES groups, peaking on day 7, accompanied by downregulation of epithelial barrier genes (Occludin, Cadherin-1, and Zo-1)(all P<0.05). Transcriptomic analysis showed that both LSS and ES activated chemokine-related pathways and enriched leukocyte migration and neutrophil recruitment pathways. Further validation revealed upregulation of CXCL2 and MMP12 in the LSS group, whereas CXCL3 and MMP12 were predominantly elevated in the ES group. CONCLUSIONS Both LSS and ES can induce sustained lung injury and neutrophil infiltration in mice, though the underlying molecular mechanisms differ. Compared with ES, exposure to LSS additionally triggers a transient eosinophilic response, suggesting that lunar dust particles possess stronger immunostimulatory potential and higher biological toxicity.
Animals ; Mice ; Mice, Inbred C57BL ; Soil ; Lung Injury/etiology* ; Dust ; Bronchoalveolar Lavage Fluid ; Moon ; Lung/pathology* ; Inhalation Exposure/adverse effects* ; Male

Yinyang Gongji Pills have the effects of strengthening the body resistance to eliminate pathogenic factors, removing stasis, and reducing swelling, which is a commonly used traditional Chinese medicine(TCM) formula for treating intestinal accumulation. A real-world, registered, and single-arm clinical trial was conducted to observe the clinical efficacy and safety of Yinyang Gongji Pills combined with capecitabine in the treatment of advanced colorectal cancer and analyze the clinical characteristics of the patients. A total of 60 patients with advanced colorectal cancer who refused or could not tolerate standard treatment of western medicine were included in the study. They were treated with Yinyang Gongji Pills combined with capecitabine until disease progression or intolerable adverse events occurred. The main observation indicators were progression-free survival(PFS) and safety. The treatment effects of the patients under different baseline characteristics were analyzed. The clinical trial has found that the median PFS of all enrolled patients was 7.3 months, with 30.1% of patients having a PFS exceeding 12.0 months. Layered analysis showed that the median PFS of patients with the onset site being the colon and rectum were respectively 8.4 and 4.7 months. The median PFS of patients with high, medium, and low tumor burden were respectively 7.0, 4.7, and 10.8 months. The median PFS of patients with wild-type and mutant-type RAS/BRAF were respectively 7.9 and 6.9 months. The median PFS of patients with KPS scores ≥80 and ≤70 were respectively 7.9 and 6.5 months. The median PFS of patients treated with Yinyang Gongji Pills for ≥6, 3-6, and ≤3 months were respectively 8.0, 5.2, and 4.2 months. The median PFS of patients with spleen, kidney, liver, and lung syndrome differentiation in TCM were respectively 8.3, 6.7, 7.3, and 5.6 months. The median PFS of patients with TCM pathological factors including phlegm, dampness, and blood stasis were respectively 7.0, 7.3, and 6.5 months. Common adverse reactions include anemia, decreased white blood cells, decreased appetite, fatigue, and hand foot syndrome, with incidence rates being respectively 44.2%, 34.6%, 42.3%, 32.7%, and 17.3%. The results showed that the combination of Yinyang Gongji Pills and capecitabine demonstrated potential clinical efficacy and good safety in this study. The patients have clinical characteristics such as low tumor burden, onset site at the colon, KPS scores ≥ 80, long duration of oral TCM, and TCM syndrome differentiation including spleen or liver.
Humans ; Capecitabine/adverse effects* ; Colorectal Neoplasms/mortality* ; Drugs, Chinese Herbal/adverse effects* ; Male ; Middle Aged ; Female ; Aged ; Adult ; Treatment Outcome

The interactions between microorganisms and medicinal plants are crucial to the quality improvement of medicinal plants. Medicinal plants attract microorganisms to colonize by secreting specific compounds and provide niche and nutrient support for these microorganisms, with a symbiotic network formed. These microorganisms grow in the rhizosphere, phyllosphere, and endophytic tissues of plants and significantly improve the growth performance and medicinal component accumulation of medicinal plants by promoting nutrient uptake, enhancing disease resistance, and regulating the synthesis of secondary metabolites. Microorganisms are also widely used in the ecological planting of medicinal plants, and the growth conditions of medicinal plants are optimized by simulating the microbial effects in the natural environment. The interactions between microorganisms and medicinal plants not only significantly improve the yield and quality of medicinal plants but also enhance their geoherbalism, which is in line with the concept of green agriculture and eco-friendly development. This study reviewed the research results on the interactions between medicinal plants and microorganisms in recent years and focused on the analysis of the great potential of microorganisms in optimizing the growth environment of medicinal plants, regulating the accumulation of secondary metabolites, inducing systemic resistance, and promoting the ecological planting of medicinal plants. It provides a scientific basis for the research on the interactions between medicinal plants and microorganisms, the research and development of microbial agents, and the application of microorganisms in the ecological planting of medicinal plants and is of great significance for the quality improvement of medicinal plants and the green and sustainable development of TCM resources.
Plants, Medicinal/metabolism* ; Bacteria/genetics* ; Symbiosis

Objective Ticks serve as vectors for transmitting Babesia microti.However,the specific mechanism remains unclear.This study aimed to investigate the effect of tick autophagy molecules on the proliferation of Babesia microti.Methods An experimental model of infected and uninfected mice was used to collect tick materials for proteomic analysis to identify differentially expressed autophagy-related molecules in Haemaphysalis longicornis.The cloning of the HlATG8 gene,protein expression,and production of polyclonal antibodies were completed.The HlATG8 gene was then knocked down using RNAi interference technology.Results The tick autophagy molecule,HlATG8,was identified and found to be significantly upregulated in ticks infected with Babesia microti.The load of Babesia microti in ticks increased significantly following the knockdown of the HlATG8 gene.Conclusions The tick autophagy molecule in Hae.longicornis,HlATG8,inhibits the proliferation of Babesia.