Zhen-Wu decoction and lactiflorin, an ingredient predicted by in silico modelling, alleviate uremia induced cardiac endothelial injury via Nrf2 activation.

ETHNOPHARMACOLOGICAL RELEVANCE: Cardiorenal syndrome type 4 (CRS type 4), with high rates of morbidity and mortality, has become a social and economic problem worldwide over the last few decades. Zhen-Wu decoction, a traditional medicine used in East Asia, has been widely used in the treatment of cardiovascular disease and kidney disease, and has shown potential therapeutic effects for the clinical treatment of CRS type 4. However, the underlying mechanism has not been extensively explored. AIM OF THE STUDY: The purpose of this study was to investigate the effect and underlying mechanism of Zhen-Wu decoction on uremic cardiomyopathy, offering a potential target for clinical treatment of CRS type 4. MATERIALS AND METHODS: Five/six nephrectomized mice were utilized for experiments in vivo. The cardioprotective effects of Zhen-Wu decoction were evaluated by echocardiography and tissue staining. RNA-Seq data were used to investigate the potential pharmacological mechanism. The prediction of targets and active components was based on our previous strategy. Subsequently, the protective effect of the selected compound was verified in experiments in vitro. RESULTS: Zhen-Wu decoction alleviated cardiac dysfunction and endothelial injury in 5/6 nephrectomized mice, and the mechanism may involve the inflammatory process and oxidative stress. The activation of the Nrf2 signaling pathway was predicted to be a potential target of Zhen-Wu decoction in protecting endothelial cells. Through our machine learning strategy, we found that lactiflorin as an ingredient in Zhen-Wu decoction, alleviates IS-induced endothelial cell injury by blocking Keap1 and activating Nrf2. CONCLUSIONS: The present study demonstrated that Zhen-Wu decoction and lactiflorin could protect endothelial cells against oxidative stress in mice after nephrectomy by activating the Nrf2 signaling pathway.

Single-cell transcriptomic analyses of cardiac immune cells reveal that Rel-driven CD72-positive macrophages induce cardiomyocyte injury.

AIMS: The goal of our study was to investigate the heterogeneity of cardiac macrophages (CMφs) in mice with transverse aortic constriction (TAC) via single-cell sequencing and identify a subset of macrophages associated with heart injury. METHODS AND RESULTS: We selected all CMφs from CD45+ cells using single-cell mRNA sequencing data. Through dimension reduction, clustering, and enrichment analyses, CD72hi CMφs were identified as a subset of pro-inflammatory macrophages. The pseudo-time trajectory and ChIP-Seq analyses identified Rel as the key transcription factor that induces CD72hi CMφ differentiation. Rel KD and Rel-/- bone marrow chimaera mice subjected to TAC showed features of mitigated cardiac injury, including decreased levels of cytokines and ROS, which prohibited cardiomyocyte death. The transfer of adoptive Rel-overexpressing monocytes and CD72hi CMφ injection directly aggravated heart injury in the TAC model. The CD72hi macrophages also exerted pro-inflammatory and cardiac injury effects associated with myocardial infarction. In humans, patients with heart failure exhibit increased CD72hi CMφ levels following dilated cardiomyopathy and ischaemic cardiomyopathy. CONCLUSION: Bone marrow-derived, Rel-mediated CD72hi macrophages play a pro-inflammatory role, induce cardiac injury and, thus, may serve as a therapeutic target for multiple cardiovascular diseases.

G-MDSCs promote aging-related cardiac fibrosis by activating myofibroblasts and preventing senescence.

Aging is one of the most prominent risk factors for heart failure. Myeloid-derived suppressor cells (MDSCs) accumulate in aged tissue and have been confirmed to be associated with various aging-related diseases. However, the role of MDSCs in the aging heart remains unknown. Through RNA-seq and biochemical approaches, we found that granulocytic MDSCs (G-MDSCs) accumulated significantly in the aging heart compared with monocytic MDSCs (M-MDSCs). Therefore, we explored the effects of G-MDSCs on the aging heart. We found that the adoptive transfer of G-MDSCs of aging mice to young hearts resulted in cardiac diastolic dysfunction by inducing cardiac fibrosis, similar to that in aging hearts. S100A8/A9 derived from G-MDSCs induced inflammatory phenotypes and increased the osteopontin (OPN) level in fibroblasts. The upregulation of fibroblast growth factor 2 (FGF2) expression in fibroblasts mediated by G-MDSCs promoted antisenescence and antiapoptotic phenotypes of fibroblasts. SOX9 is the downstream gene of FGF2 and is required for FGF2-mediated and G-MDSC-mediated profibrotic effects. Interestingly, both FGF2 levels and SOX9 levels were upregulated in fibroblasts but not in G-MDSCs and were independent of S100A8/9. Therefore, a novel FGF2-SOX9 signaling axis that regulates fibroblast self-renewal and antiapoptotic phenotypes was identified. Our study revealed the mechanism by which G-MDSCs promote cardiac fibrosis via the secretion of S100A8/A9 and the regulation of FGF2-SOX9 signaling in fibroblasts during aging.

Diagnostic Accuracy of the Synovial Fluid α-Defensin Lateral Flow Test in Periprosthetic Joint Infection: A Meta-analysis.

OBJECTIVES: There is a controversy on the diagnostic reliability and accuracy of synovial fluid α-defensin in periprosthetic joint infection (PJI). We performed this meta-analysis to evaluate the diagnostic accuracy of the α-defensin lateral flow test in PJI. METHODS: PubMed, Embase, and the Cochrane library were systematically searched, and articles (up to January 2020) on the diagnosis of hip and knee PJIs using the α-defensin Synovasure lateral flow test were included. The diagnostic accuracy of the α-defensin lateral flow test in PJI was evaluated using meta-analysis. The pooled sensitivity, specificity, accuracy, positive and negative likelihood ratio, diagnostic odds ratio, and post-test probabilities were calculated. RESULTS: Seventeen studies including 1443 cases were included. Meta-analysis showed the pooled sensitivity, specificity, positive likelihood ratio, negative likelihood ratio, and a diagnostic odds ratio was 0.83 (95% CI 0.77, 0.88), 0.95 (95% CI 0.93, 0.97), 16.86 (95% CI 11.67, 24.37), 0.17 (95% CI 0.13, 0.24) and 85.30 (95% CI 47.76, 152.35), respectively. The area under the hierarchical summary receiver operating characteristic curve was 0.97 (95% CI 0.95, 0.98). Subgroup analysis also confirmed the high efficiency of α-defensin Synovasure lateral flow test in diagnosing PJIs, irrespective of ethnicity. Fagan's nomogram analysis there was a high positive post-test probability of 94% and a low negative post-test probability of 15%. CONCLUSIONS: We indicated that the α-defensin lateral flow test had a high accuracy for diagnosing PJI. Large-scale studies are needed to validate its significance in PJI diagnosis.

[Advances on treatment of periprosthetic infection and antibiotic delivery system after total hip arthroplasty].

Periprosthetic infection after hip replacement is a clinical catastrophic disease, which often leads to the failure of the prosthesis. It needs the combination of systemic antibiotics to cure the infection, which brings huge burden to doctors and patients. There are strict indications for debridement and one-stage revision of the prosthesis, and few cases meet the requirements. The second revision is still the gold standard for the treatment of periprosthetic infection. It is suitable for all infection conditions and has a high success rate. On the second phase of renovation, the antibiotic sustained release system plays a key role, and the carrier of antibiotic sustained-release system is the focus of current research, including classic bone cement and absorbable biomaterials. Bone cement has strong mechanical strength, but the antibiotic release shows a sharp decline trend; the absorbable biomaterials can continuously release antibiotics with high concentration, but the mechanical strength is poor, so it could not use alone. The combination of bone cement and absorbable biomaterials will be an ideal antibiotic carrier. PMMA is the most commonly used antibiotic carrier, but the antibiotic release concentration is decreased sharply after 24 hours. It will be difficult to control the infection and increase the risk of bacterial resistance if it is lower than the minimum inhibitory concentration. The biodegradable materials can release antibiotics completely, with long release time and high concentration, but low mechanical strength. Antibiotic spacer plays an important role in the control of infection. In the future, how to further extend the antibiotic release time of antibiotic sustained-release system, increase the amount of antibiotic release and maintain the mechanical strength of the material will be studied.

[Procalcitonin as a sensitive index of chronic infection after artificial joint replacement:a retrospective analysis].

OBJECTIVE: To study the predictive value of procalcitonin as a serum biomarker in diagnosis of late periprosthetic joint infection(PJI) for providing theoretical reference basis for diagnosis of PJI. METHODS: A total of 77 cases were retrospective included from January 2015 to December 2017 for revision of total hip arthroplasty and total knee arthroplasty, according to the diagnostic criteria of Musculoskeletal Infection Society(MISI). All cases were divided into infection group and non-infection group. Infection group included 21 cases, 7 cases for male and 14 cases for female, with an average age of (60.70±8.75) years old (ranged 43 to 75 years old). Non-infection group 56 included cases, 24 cases for male and 32 cases for female, with an average age (64.40±12.14) years old (ranged 43 to 85 years old). Concentration of preoperative serum procalcitonin was examined and the chi-square test was used to compare positive rate between the two groups. RESULTS: Two cases in the infection group had positive serum procalcitonin, 0.06 ng/L and 0.10 ng/L respectively, with the positive rate of 9.52%; 4 cases in non-infected group had positive serum procalcitonin, 0.05 ng/L, 0.06 ng/L, 0.06 ng/L, 0.16 ng/L respectively, with the positive rate of 7.14%. No statistically significant difference was observed between the two groups (P=0.662). CONCLUSIONS: Most of PJI were low toxicity infection with little systemic inflammatory response, so the concentration of serum procalcitonin was normal or slightly higher level, had little clinical significance for diagnosis of PJI. But the cases of this retrospective study are not enough, more cases are needed for further study.

Hispaglabridin B, a constituent of liquorice identified by a bioinformatics and machine learning approach, relieves protein-energy wasting by inhibiting forkhead box O1.

BACKGROUND AND PURPOSE: Liquorice is the root of Glycyrrhiza glabra, which is a popular food in Europe and China that has previously shown benefits for skeletal fatigue and nutrient metabolism. However, the mechanism and active ingredients remain largely unclear. The aim of this study was to investigate the active ingredients of liquorice for muscle wasting and elucidate the underlying mechanisms. EXPERIMENTAL APPROACH: RNA-Seq and bioinformatics analysis were applied to predict the main target of liquorice. A machine learning model and a docking tool were used to predict active ingredients. Isotope labelling experiments, immunostaining, Western blots, qRT-PCR, ChIP-PCR and luciferase reporters were utilized to test the pharmacological effects in vitro and in vivo. The reverse effects were verified through recombination-based overexpression. KEY RESULTS: The liposoluble constituents of liquorice improved muscle wasting by inhibiting protein catabolism and fibre atrophy. We further identified FoxO1 as the target of liposoluble constituents of liquorice. In addition, hispaglabridin B (HB) was predicted as an inhibitor of FoxO1. Further studies determined that HB improved muscle wasting by inhibiting catabolism in vivo and in vitro. HB also markedly suppressed the transcriptional activity of FoxO1, with decreased expression of the muscle-specific E3 ubiquitin ligases MuRF1 and Atrogin-1. CONCLUSIONS AND IMPLICATIONS: HB can serve as a novel natural food extract for preventing muscle wasting in chronic kidney disease and possibly other catabolic conditions.

FIBP knockdown attenuates growth and enhances chemotherapy in colorectal cancer via regulating GSK3ß-related pathways.

Colorectal cancer stem cells (CSCs), characterized by self-renewal ability and high expression of proliferative genes, contribute to the chemoresistance of colorectal cancer (CRC). We aimed to identify the molecular mechanisms underlying CRC chemoresistance through comprehensive bioinformatics screenings and experimental confirmation of gene functions. We found that high expression of FGF1 intracellular binding protein (FIBP) was correlated with chemoresistance and poor prognosis in CRC patients. Therefore, the chemoresistant CRC cell line HCT116-CSC with high expression of the stem cell markers CD44 and CD133 was established for further phenotypic tests. FIBP knockdown inhibited proliferation, enhanced chemotherapy effects, and attenuated the stemness markers of CRC cells in vivo and in vitro. Through RNA-seq and gene set enrichment analysis, we identified cyclin D1 as a key downstream target in FIBP-regulated cell cycle progression and proliferation. Moreover, FIBP bound to GSK3ß, inhibited its phosphorylation at Tyr216, and activated ß-catenin/TCF/cyclin D1 signaling in HCT116-CSCs. Additional GSK3ß knockdown reversed the FIBP silencing-induced inhibition of proliferation and decreased stemness marker expression in HCT116-CSCs. Furthermore, DNA methylation profiling suggested that FIBP regulated the stemness of CRC cells via methylation activity that was dependent on GSK3ß but independent of ß-catenin signaling. Our data illuminate the potential of FIBP as a novel therapeutic target for treating chemoresistant CRC through inhibition of GSK3ß-related signaling.

The p.R229Q variant of the NPHS2 (podocin) gene in focal segmental glomerulosclerosis and steroid-resistant nephrotic syndrome: a meta-analysis.

While many previous studies have reported an association between the p.R229Q variant of the NPHS2 gene and focal segmental glomerulosclerosis (FSGS) or steroid-resistant nephrotic syndrome (SRNS), a conclusive relationship has not been defined. In this study, we performed a meta-analysis of the published data to investigate the impact of the p.R229Q polymorphism on FSGS and SRNS patients. Despite significant heterogeneity within some of the comparisons, the results revealed significantly higher risks of SRNS in individuals homozygous for the variant allele (OR 7.411, 95% confidence interval 1.876-29.436, p = 0.004) compared to homozygous non-variant individuals. However, the carrier rate of the p.R229Q variant was not significantly different between SRNS patients and steroid-sensitive nephrotic syndrome patients. No statistically significant differences in the p.R229Q carrier rate were observed between FSGS patients and controls or FSGS patients and patients with different pathology classifications. No notable differences in the p.R229Q carrier rate were found between patients and controls in any group with early-onset disease (onset age < 18). In conclusion, our meta-analysis suggests that for adult-onset disease (onset age > 18), the homozygous variant could be a potential predictor of hereditary nephrotic syndrome and that the p.R229Q allele cannot currently be considered a risk factor for predicting FSGS.

Vascularised greater trochanter bone graft, combined free iliac flap and impaction bone grafting for osteonecrosis of the femoral head.

PURPOSE: To investigate the curative efficacy of osteonecrosis of the femoral head (ONFH) in a hip-preserving operative approach, by grafting a vascularized greater trochanter flap combined with a free iliac flap, in an attempt to seek an innovative approach for patients who suffered middle to late stage ONFH without total hip arthroplasty (THA) surgery. METHOD: Our research included a total of 60 patients (66 hips) who accepted hip-preserving surgery by grafting a vascularized greater trochanter flap combined with a free iliac flap which was tightly filled by hammering because of ONFH (most were Association Research Circulation Osseous (ARCO) stage III patients) from January, 2006 to December, 2010. A Harris Hip Score was obtained during follow-ups, evaluating the clinical efficacy, X-rays were taken regularly for image assessing, and the SF-36 scale was used for estimating quality of life. Terminal observation time was considered when patients had symptom-dependant indications for performing another hip-preserving surgery or THA surgery. RESULTS: Fifty-eight patients (64 hips) were eventually contacted by telephone for an out-patient clinic return visit, with a mean follow-up time of 35.8 months (varied from 12 months to 60 months), but two patients lost contact for various reasons. The demographic data were as follows: there were 16 ARCO IIIA cases, 22 ARCO IIIB cases, and 26 ARCO IIIC cases, respectively. Postoperative X-rays revealed a well-repaired necrotic area of the femoral head and improvement of femoral-acetabulum coverage. The last follow-up mean Harris Hip Score was 86.56 ± 7.38 (excellent results reached 87.50%), which were greatly improved compared to 50.95 ± 6.86 pre-operatively. Also the postoperative mean scores of all dimensions of the SF-36 scale were improved to some extent. Additionally the physical component summary (PSC) scores were enhanced from 42 ± 13 pre-operatively to 78 ± 11, while the postoperative mental component summary (MCS) scores (76 ± 11) largely increased in contrast to pre-operative scores (51 ± 10), with both target indices having statistical significance (p = 0.005, p = 0.01), signifying hugely improvement of the quality of life of the patients. A correlation was found between Harris Hip Score and all dimensions of SF-36 scale (r = 0.32-0.72), especially closely correlated with physical functioning (PF), role-physical (RP) and bodily pain (BP) in PCS aspect (r = 0.72, p < 0.01; r = 0.58, p < 0.01; r = 0.65, p < 0.01, respectively). CONCLUSION: There is definite curative efficacy for the treatment of ONFH with an hip-preserving operative approach by grafting a vascularized greater trochanter flap combined with a free iliac flap which was tightly filled by hammering. This kind of operative approach reconstructs the biological stability of femoral head, which promotes repair of necrotic areas and indirectly preserves the femoral head of patients and a majority of hip function. It possesses vast clinical as well as practical significance, because the long-term efficacy can satisfy fundamental life requirements, especially for those young and middle-aged patients who suffer ONFH to avoid or put off the time of total hip arthroplasty (THA) surgery.