Assessment of the relationship between gut microbiota and bone mineral density: a two-sample Mendelian randomization study.

Background: Emerging evidence from observational studies and clinical trials suggests a connection between the gut microbiota and variations in bone mineral density (BMD). Nonetheless, the specific association between gut microbiota and BMD alterations at different skeletal sites has not been comprehensively explored. To address this, we employed Genome-Wide Association Study (GWAS) summary statistics from a publicly accessible database, conducting a two-sample Mendelian Randomization analysis to elucidate the potential causal relationship between gut microbiota composition and BMD. Methods: This study utilized two distinct thresholds for screening instrumental variables (IVs), followed by an extensive series of quality control procedures to identify IVs that were significantly related to exposure. Gut microbiota were classified into two sets based on hierarchical levels: phylum, class, order, family, and genus. Bone mineral density (BMD) data were systematically collected from four skeletal sites: femoral neck, lumbar spine, forearm, and heel. For Mendelian Randomization (MR) analysis, robust methods including Inverse-Variance Weighting (IVW) and the Wald Ratio Test were employed. Additional analytical tests such as the Outlier Test, Heterogeneity Test, 'Leave-One-Out' Test, and Pleiotropy Test were conducted to assess the impact of horizontal pleiotropy, heterogeneities, and the genetic variation stability of gut microbiota on BMD causal associations. The MR Steiger Directionality Test was applied to exclude studies with potential directional biases. Results: In this two-sample Mendelian randomization analysis, we utilized five sets of exposure GWAS (Genome-Wide Association Studies) summary statistics and four sets of outcome GWAS summary statistics. The initial analysis, applying a threshold of p < 5 × 10-6, identified 48 significant causal relationships between genetic liability in the gut microbiome and bone mineral density (BMD). A subsequent analysis with a more stringent threshold of p < 5 × 10-8 uncovered 14 additional causal relationships. Upon applying the Bonferroni correction, 9 results from the first analysis and 10 from the second remained statistically significant. Conclusion: Our MR analysis revealed a causal relationship between gut microbiota and bone mineral density at all sites, which could lead to discoveries in future mechanistic and clinical studies of microbiota-associated osteoporosis.

[Anti-tumor mechanism of total saponins of Paridis Rhizoma on inducing ferroptosis of breast cancer MCF-7 cells].

This study aims to investigate the mechanism of total saponins of Paridis Rhizoma in inducing the ferroptosis of MCF-7 cells and provide a theoretical basis for the clinical treatment of breast cancer with total saponins of Paridis Rhizoma. The methyl thiazolyl tetrazolium(MTT) assay was employed to examine the effects of different concentrations of total saponins of Paridis Rhizoma on the proliferation of MCF-7 cells. A phase contrast inverted microscope was used to observe the morphological changes of MCF-7 cells. The colony formation assay was employed to test the colony formation of MCF-7 cells. The lactate dehydrogenase(LDH) release test was conducted to determine the cell membrane integrity of MCF-7 cells. The cell scratch assay was employed to examine the migration of MCF-7 cells. After that, the level of reactive oxygen species(ROS) in MCF-7 cells was observed by an inverted fluorescence microscope, and the content of Fe~(2+) in MCF-7 cells was detected by the corresponding kit. Transmission electron microscopy was employed to observe the mitochondrial ultrastructure of MCF-7 cells. Western blot was employed to determine the expression of ferroptosis-related proteins, such as p53, solute carrier family 7 member 11(SLC7A11), glutathione peroxidase 4(GPX4), acyl-CoA synthetase long-chain family member 4(ACSL4), and transferrin receptor protein 1(TFR1) in MCF-7 cells. The results showed that 1.5, 3, 4.5, 6, 7.5, and 9 µg·mL~(-1) total saponins of Paridis Rhizoma significantly inhibited the proliferation of MCF-7 cells, with the IC_(50) of 4.12 µg·mL~(-1). Total saponins of Paridis Rhizoma significantly damaged the morphology of MCF-7 cells, leading to the formation of vacuoles and the gradual shrinkage and detachment of cells. Meanwhile, total saponins of Paridis Rhizoma inhibited the colony formation of MCF-7 cells, destroyed the cell membrane(leading to the release of LDH), and shortened the migration distance of MCF-7 cells. Total saponins of Paridis Rhizoma treatment significantly increased the content of ROS, induced oxidative damage, and led to the accumulation of Fe~(2+) in MCF-7 cells. Furthermore, total saponins of Paridis Rhizoma changed the mitochondrial structure, increased the mitochondrial membrane density, led to the decrease or even disappear of ridges, promoted the expression of p53 protein, down-regulated the expression of SLC7A11 and GPX4, and up-regulated the expression of ACSL4 and TFR1. In summary, total saponins of Paridis Rhizoma can significantly inhibit the proliferation and migration of MCF-7 cells and destroy the cell structure by inducing ferroptosis.

Enhanced lysosomal escape of cell penetrating peptide-functionalized metal-organic frameworks for co-delivery of survivin siRNA and oridonin.

In recent years, small interfering RNA (siRNA) has been widely used in the treatment of human diseases, especially tumors, and has shown great appeal. However, the clinical application of siRNA faces several challenges. Insufficient efficacy, poor bioavailability, poor stability, and lack of responsiveness to a single therapy are the main problems affecting tumor therapy. Here, we designed a cell-penetrating peptide (CPP)-modified metal organic framework nanoplatform (named PEG-CPP33@ORI@survivin siRNA@ZIF-90, PEG-CPP33@NPs) for targeted co-delivery of oridonin (ORI), a natural anti-tumor active ingredient) and survivin siRNA in vivo. This can improve the stability and bioavailability of siRNA and the efficacy of siRNA monotherapy. The high drug-loading capacity and pH-sensitive properties of zeolite imidazolides endowed the PEG-CPP33@NPs with lysosomal escape abilities. The Polyethylene glycol (PEG)-conjugated CPP (PEG-CPP33) coating significantly improved the uptake in the PEG-CPP33@NPs in vitro and in vivo. The results showed that the co-delivery of ORI and survivin siRNA greatly enhanced the anti-tumor effect of PEG-CPP33@NPs, demonstrating the synergistic effect between ORI and survivin siRNA. In summary, the novel targeted nanobiological platform loaded with ORI and survivin siRNA presented herein showed great advantages in cancer therapy, and provides an attractive strategy for the synergistic application of chemotherapy and gene therapy.

A comparative study of the efficacy of intra-articular injection of different drugs in the treatment of mild to moderate knee osteoarthritis: A network meta-analysis.

BACKGROUND: Knee osteoarthritis (KOA) has a high clinical prevalence and frequently interferes with patients normal lives. In KOA patients, evidence suggests that intra-articular (IA) injection improves joint function and decreases discomfort. Several IA injection treatments are used in daily practice to improve symptomatic control of knee osteoarthritis, but their efficacy is frequently disputed. METHODS: This network meta-analysis compares the efficacy of different IA injections for mild to moderate knee osteoarthritis. Seven databases (PubMed, EMBASE, Web of Science, Cochrane Library, China Biology Medicine disc, WanFang, and China National Knowledge Infrastructure) were searched for randomized controlled trials published up to and including December 20, 2021, and final follow up indicators were used. Visual analogue scale (VAS) score and The Western Ontario and McMaster Universities (WOMAC) Osteoarthritis Index (WOMAC) score change from baseline were the primary outcomes. We used the Cochrane risk of bias tool to assess the quality and risks of biases of papers. We calculated the weighted mean difference (WMD) and 95% confidence interval (CI) for each outcome. State (Version 15.1, Texas, USA) and SPSS (Version 20, Chicago, USA) was used in all statistical analyses, and Review Manager (version 5.4) was used in assessing the risks of biases. RESULTS: Our study included 16 randomized controlled trials with a total of 1652 patients. platelet-rich plasma (PRP) IA injection therapy had the highest likelihood of being the best intervention in reducing WOMAC pain (surface under the cumulative ranking area [SUCRA] 84.7%), stiffness (SUCRA 95.1%), and function (SUCRA 98.5%) scores, according to the SUCRA. The best measures for lowering the WOMAC total and VAS scores were IA injection platelet-rich plasma-derived growth factor (SUCRA 84.9%) and hyaluronic acid and platelet-rich plasma (SUCRA 84.9%). In the VAS score group, PRP outperformed hyaluronic acid (HA) (WMD 1.3, 95% CI 0.55-2.55) and corticosteroids (CS) (WMD 4.85, 95% CI 4.02-5.08), according to the forest map results. PRP also outperformed CS (WMD 14.76, 95% CI 12.11-17.41), ozone (WMD 9.16, 95% CI 6.89-11.43), and PRP + HA (WMD 2.18, 95% CI 0.55-3.81) in the WOMAC total score group. Furthermore, PRP outperforms other drugs in terms of reducing WOMAC function, stiffness, and function score. CONCLUSION: In patients with mild to moderate KOA, IA injection PRP outperformed IA injection ozone, HA, CS, platelet-rich plasma-derived growth factor, and hyaluronic acid and platelet-rich plasma in terms of pain, stiffness, and dysfunction.