Type 2 diabetes mellitus and the risk of male infertility: a Mendelian randomization study.

Objective: To assess the causal effect of type 2 diabetes mellitus (T2DM) on male infertility (MI) and erectile dysfunction (ED) by Mendelian randomization (MR) analysis. Methods: Data for T2DM, MI, and ED were obtained from genome-wide association studies (GWAS) involving 298, 957, 73, 479, and 223, 805 Europeans, respectively. We performed univariate MR analysis using MR Egger, Weighted median (WM) and Inverse variance weighted (IVW) methods to assess causal effects among the three. Through the Genotype Tissue Expression (GTEx) database, single-nucleotide polymorphisms (SNPs) that affect the expression levels of T2DM-related genes were located using expression quantitative trait loci (eQTL). Results: MR analysis showed a significant causal relationship between T2DM and ED (WM, OR: 1.180, 95%CI: 1.010-1.378, P = 0.037; IVW, OR: 1.190, 95%CI: 1.084-1.300, P < 0.001). There is also a significant causal relationship between T2DM and MI (MR Egger, OR: 0.549, 95%CI: 0.317-0.952, P = 0.037; WM, OR: 0.593, 95%CI: 0.400, P = 0.010; IVW, OR: 0.767, 95%CI: 0.600-0.980, P = 0.034). ED may not cause MI (P > 0.05). We also found that rs6585827 corresponding to the PLEKHA1 gene associated with T2DM is an eQTL variant affecting the expression of this gene. Conclusion: T2DM has a direct causal effect on ED and MI. The level of PLEKHA1 expression suppressed by rs6585827 is potentially associated with a lower risk of T2DM.

Astaxanthin prevents osteoarthritis by blocking Rspo2-mediated Wnt/ß-catenin signaling in chondrocytes and abolishing Rspo2-related inflammatory factors in macrophages.

Chondrocyte degeneration and classically activated macrophage (AM)-related inflammation play critical roles in osteoarthritis (OA). Here, we explored the effects of astaxanthin and Rspo2 on OA in vitro and in vivo. We observed that the Rspo2 gene was markedly elevated in synovial tissues of OA patients compared with healthy controls. In 2D cultures, Rspo2 and inflammatory factors were enhanced in AMs compared with nonactivated macrophages (NMs), and the protein expression levels of Rspo2, ß-catenin, and inflammatory factors were increased, and anabolic markers were reduced in osteoarthritic chondrocytes (OACs) compared to normal chondrocytes (NCs). Astaxanthin reversed these changes in AMs and OACs. Furthermore, Rspo2 shRNA significantly abolished inflammatory factors and elevated anabolic markers in OACs. In NCs cocultured with AM, and in OACs cocultured with AMs or NMs, astaxanthin reversed these changes in these coculture systems and promoted secretion of Rspo2, ß-catenin and inflammatory factors and suppressed anabolic markers compared to NCs or OACs cultured alone. In AMs, coculture with NCs resulted in a slight elevation of Rspo2 and AM-related genes, but not protein expression, compared to culture alone, but when cocultured with OACs, these inflammatory mediators were significantly enhanced at both the gene and protein levels. Astaxanthin reversed these changes in all the groups. In vivo, we observed a deterioration in cartilage quality after intra-articular injection of Rspo2 associated with medial meniscus (DMM)-induced instability in the OA group, and astaxanthin was protective in these groups. Our results collectively revealed that astaxanthin attenuated the process of OA by abolishing Rspo2 both in vitro and in vivo.

Establishment and validation of haematological reference intervals for newborns aged 5 to 28 days in Nanjing, China.

BACKGROUND: China lacks a standard for reference intervals (RIs) of complete blood cell (CBC) counts in newborns. This study aimed to determine local haematological RIs for newborns. METHODS: This prospective study was conducted from January 2020 to December 2020 in Nanjing Maternity and Child Health Care Hospital. We collected capillary blood specimens from 497 healthy newborns aged 5-28 days. We calculated the RIs as nonparametric 2.5th to 97.5th percentiles and 90% confidence intervals following the EP28-A3c guideline. We validated the RIs in another 20 specimens from healthy newborns. RESULTS: The RIs for the 18 CBC parameters were: white blood cell count 7.17-15.69 × 109/L; monocytes# 0.52-1.66 × 109/L; Mono% 5.7-14.1; eosinophils# 0.16-1.08 × 109/L; Eos% 1.5-8.9; Lymphocytes# 4.04-8.08 × 109/L; Lymph% 40.1-67.8; Neutrophils# 1.59-6.41 × 109/L; Neut% 18.1-46.7; Basophils# 0.00-0.04 × 109/L; Baso% 0.0-0.4; red blood cell count 3.38-5.89 × 1012/L; haemoglobin (Hb) 116-198 g/L; Hematocrit % 35.2-61.2; mean corpuscular Hb (MCH) 30.9-36.3 pg/L; MCH concentration 310-341 g/L; mean corpuscular volume 94.5-112.2 fL; and platelets 210-610 × 109/L. The RIs had a conformity rate of 90%-100% in the validation specimens. CONCLUSION: We established region-specific RIs for CBC parameters in healthy newborns to help diagnose haematological disease in neonates.

Single-cell transcriptomic data reveal the increase in extracellular matrix organization and antigen presentation abilities of fibroblasts and smooth muscle cells in patients with pelvic organ prolapse.

INTRODUCTION AND HYPOTHESIS: We aimed to explore the cellular properties of fibroblasts and smooth muscle cells (SMCs), the two major cell types of the vagina wall, in pelvic organ prolapse (POP) to improve the knowledge of the underlying molecular mechanisms of POP. METHODS: The single-cell RNA sequencing (scRNA-seq) profile GSE151202 was downloaded from NCBI Gene Expression Omnibus, in which vaginal wall tissues were harvested from patients with anterior vaginal wall prolapse and control subjects respectively. The scRNA-seq data of samples (5 POP and 5 controls) were adopted for analysis. Cluster analysis was performed to identify the cell subclusters. Trajectory analysis was applied to construct the differentiation trajectories of fibroblasts and SMCs. Cellular communication analysis was carried out to explore the ligand-receptor interactions between fibroblasts/SMCs and immune cells. RESULTS: Ten subclusters were determined in both groups, among which fibroblasts and SMCs were the most abundant cell types. Compared with controls, fibroblasts increased whereas SMCs declined in POP. During the transition of fibroblasts and SMCs from a normal into a disease state, extracellular matrix organization and antigen presentation were heightened. The intercellular communications were altered in POP. Interactions between fibroblasts/SMCs and macrophages/natural killer/T cells were strengthened as more ligand-receptor pairs involved in antigen presentation pathways were gained in POP. CONCLUSION: Extracellular matrix organization and antigen presentation abilities of fibroblasts and SMCs were enhanced in POP.

Obesity-induced oxidative stress and mitochondrial dysfunction negatively affect sperm quality.

Obesity is a systemic metabolic disease that can induce male infertility or subfertility through oxidative stress. The aim of this study was to determine how obesity impairs sperm mitochondrial structural integrity and function, and reduces sperm quality in both overweight/obese men and mice on a high-fat diet (HFD). Mice fed the HFD demonstrated higher body weight and increased abdominal fat content than those fed the control diet. Such effects accompanied the decline in antioxidant enzymes, such as glutathione peroxidase (GPX) and catalase and superoxide dismutase (SOD) in testicular and epidydimal tissues. Moreover, malondialdehyde (MDA) content significantly increased in sera. Mature sperm in HFD mice demonstrated higher oxidative stress, including increased mitochondrial reactive oxygen species (ROS) levels and decreased protein expression of GPX1, which may impair mitochondrial structural integrity and reduce mitochondrial membrane potential (MMP) and ATP production. Moreover, cyclic AMPK phosphorylation status increased, whereas sperm motility declined in the HFD mice. Clinical studies demonstrated that being overweight/obese reduced SOD enzyme activity in the seminal plasma and increased ROS in sperm, accompanied by lower MMP and low-quality sperm. Furthermore, ATP content in the sperm was negatively correlated with increases in the BMI of all clinical subjects. In conclusion, our results suggest that excessive fat intake had similar disruptive effects on sperm mitochondrial structure and function, as well as oxidative stress levels in humans and mice, which in turn induced lower sperm motility. This agreement strengthens the notion that fat-induced increases in ROS and impaired mitochondrial function contribute to male subfertility.

Serum lipidomic profiling by UHPLC-MS/MS may be able to detect early-stage endometrial cancer.

Nowadays, screening for endometrial cancer (EC) primarily relies on clinical symptoms and imaging, which makes it difficult to detect early-stage disease. Here, we conducted a widely targeted lipidomic analysis of 38 human serum samples in a discovery set and 40 human serum samples in a validation set to profile the dysregulated lipid species and establish lipid biomarkers for early-stage EC. This comprehensive lipidomic determination of 616 serum lipids indicated significant differences between early-stage EC patients and healthy controls. Three phases of lipid biomarker investigation (discovery, validation, and determination of the lipid biomarker panel) were performed, which revealed the upregulation of some sphingolipid, glycerophospholipid, and glycerolipids and downregulation of some carnitine. Consistently, the perturbation of sphingolipid and glycerophospholipid metabolism was also observed from pathway enrichment analysis. Moreover, a lipid biomarker panel, including ursodeoxycholic acid, PC(O-14:0_20:4), and Cer(d18:1/18:0), was established. This panel was assessed as an effective diagnostic model to distinguish early-stage EC patients from healthy controls and atypical endometrial hyperplasia patients within the area under the receiver operating characteristic curve (AUC) reaching 0.903 and 0.928, respectively. In particular, the comparison results of the diagnostic efficacy indicated that the lipid biomarker panel was superior to clinically established indicators for EC diagnosis, including HE4, CA125, CA153, and CA199, suggesting that it could be used as an excellent supplementary method for the diagnosis of early-stage EC. In conclusion, we established a novel and non-invasive lipid biomarker for early-stage EC detection and these findings may provide new insight into the pathological mechanisms of EC.

Optimal surgical treatment for periprosthetic distal femoral fractures after total knee arthroplasty: a Bayesian-based network analysis.

BACKGROUND: The surgical methods for periprosthetic distal femoral fractures (PDFFs) after total knee arthroplasty included locking compression plate (LCP), retrograde intramedullary nailing (RIMN), and distal femoral replacement (DFR). However, the optimal treatment remains controversial. We performed a network meta-analysis (NMA) to provide the optimal surgical method for PDFFs. MATERIALS AND METHODS: Electronic databases, including Embase, Web of Science, Cochrane Library, and PubMed, were searched for studies that compared LCP, RIMN, and DFR for PDFFs. The quality of the included studies was assessed according to the Newcastle-Ottawa scale. Pairwise meta-analysis was performed by Review Manager version 5.4. The NMA was conducted in Aggregate Data Drug Information System software version 1.16.5. We calculated odds ratios (ORs) and 95% confidence intervals (CIs) for postoperative complications and reoperations. RESULTS: A total of 19 studies and 1198 patients were included, of whom 733 for LCP, 282 for RIMN, and 183 for DFR. Pairwise meta-analysis comparing LCP to RIMN and LCP to DFR showed no significant difference in complications and reoperations except that RIMN had a higher risk of malunion comparing to LCP (OR 3.05; 95% CI 1.46-6.34; P = 0.003). No statistically significant effects were found in the NMA of overall complications, infection, and reoperation. However, results of rank probabilities showed that DFR ranked best in overall complications and reoperation, RIMN ranked best in infection but worst in reoperation, and LCP ranked worst in infection and middle in reoperation. DISCUSSION: We found similar complication rate and reoperation rate between LCP, RIMN, and DFR. The results of rank probabilities favored DFR, and further studies with high-level evidence are expected to verify the optimal surgical method for PDFFs. LEVEL OF EVIDENCE: Level II; network meta-analysis.

Exosomal miR-374c-5p derived from mesenchymal stem cells suppresses epithelial-mesenchymal transition of hepatocellular carcinoma via the LIMK1-Wnt/ß-catenin axis.

Metastasis is a leading cause to treatment failure in hepatocellular carcinoma (HCC) patients. Exosomes act as pivotal mediators in communication between different cells and exert effects on recipient cells by delivering bioactive cargoes, such as microRNAs (miRNAs). MiRNAs function in multiple steps of HCC development, including metastasis. MiR-374c-5p was previously identified as a tumor suppressor in some malignancies, while the current knowledge of its role in HCC metastasis is still limited. Herein, miR-374c-5p was found to be downregulated in HCC cell lines and clinical samples, and positively related with favorable prognosis in HCC patients. MiR-374c-5p transferred by exosomes derived from bone marrow mesenchymal stem cell (BMSC) suppressed migration, invasion and proliferation of HCC cells. LIMK1 was verified as downstream target gene of miR-374c-5p. Knockdown of LIMK1 reduced invasion, migration and proliferation of HCC cells, whereas overexpression functioned oppositely. The miR-374c-5p/LIMK1 axis suppressed epithelial-mesenchymal transition (EMT) by inactivating Wnt/ß-catenin pathway. In addition, miR-374c-5p was downregulated and LIMK1 upregulated in TGF-ß1 induced EMT. This EMT model could be reversed by LIMK1 silencing or miR-374c-5p overexpression. These results suggest that exo-miR-374c-5p suppresses EMT via targeting LIMK1-Wnt/ß-catenin axis and the axis is involved in TGF-ß1 induced metastasis of HCC, thereby identifying miR-374c-5p as a potential target for HCC treatment.

PAK2 is essential for chromosome alignment in metaphase I oocytes.

As a highly conserved and ubiquitously expressed serine/threonine kinase, p21-activated kinase 2 (PAK2) participates in diverse biologic events. However, its roles in mouse oocyte meiotic maturation remain unclear. The present study revealed that mouse oocytes depleted of Pak2 were unable to completely progress through meiosis and that a majority were arrested at metaphase I. Pak2 depletion thus prompted MI arrest and induced meiotic chromosome alignment defects in mouse oocytes, in part due to a reduction in polo-like kinase (PLK1). We demonstrated that PAK2's interaction with PLK1 protected it from degradation by APC/CCdh1, and that it promoted meiotic progression and bipolar spindle formation. Our data collectively display critical functions for PAK2 in meiotic progression and chromosome alignment in mouse oocytes.

Baicalein limits osteoarthritis development by inhibiting chondrocyte ferroptosis.

Osteoarthritis (OA) is a common degenerative disease of articular cartilage, and there is currently no effective treatment. Baicalein, a flavonoid extracted from plants of the Scutellaria genus, has frequently been used in the past as an anti-inflammatory and anti-allergic therapy. This study investigated the effect of baicalein on OA development. For in vivo study, a destabilization of the medial meniscus (DMM)-induced OA model was established in 8-week-old wild-type (WT) and AMPKα-knockout (KO) mice, while an in vitro study was performed using chondrocytes in an OA microenvironment induced by interleukin-1ß (IL-1ß) exposure. We found that baicalein alleviated OA development in vivo and exerted a chondroprotective effect in vitro by suppressing chondrocyte ferroptosis. Baicalein reduced OA-related pain sensitivity by inhibiting ferroptosis of chondrocytes in OA mice. Baicalein also facilitated AMPK holoenzyme assembly, stability, and activity and suppressed ferroptosis by inducing AMPKα phosphorylation in chondrocyte. In addition, AMPKα preserved nuclear factor erythroid 2-related factor 2(Nrf2) abundance in chondrocytes and induced Nrf2 into nucleus by promoting Keap1 degradation. Meanwhile, Nrf2 increased expression of heme oxygenase-1(HO-1) to inhibit chondrocyte lipid ROS. Taken together, these results showed that baicalein alleviated OA development by improving the activity of AMPK/Nrf2/HO-1 signaling to inhibit chondrocyte ferroptosis, revealing baicalein to be a potential therapeutic strategy for OA.

Comparison of locking compression plate and distal femoral replacement for periprosthetic distal femoral fractures: a retrospective study.

OBJECTIVE: To compare the clinical outcomes of locking compression plate (LCP) and distal femoral replacement (DFR) for periprosthetic distal femoral fractures (PDFFs) after total knee arthroplasty. METHODS: This retrospective study identified patients with PDFFs in an institutional database between January 2012 and December 2021. Demographic data and clinical outcomes, including postoperative complications, reoperation, 1-year mortality, Knee Society Scores (KSS) and Knee Society Functional Score (KSFS) were analysed. RESULTS: In total, 12 patients treated with LCP and six patients treated with DFR were included. There was no significant difference between the LCP and DFR groups in terms of postoperative complications (25.0% versus 33.3%, respectively), reoperation (8.3% versus 0.0%), respectively, 1-year mortality (8.3% versus 16.7%, respectively) or mean ± SD KSS (80.3 ± 8.3 versus 78.0 ± 2.5, respectively). However, the mean ± SD KSFS was significantly better for patients with LCP than for those with DFR (51.8 ± 12.5 versus 37.0 ± 6.7, respectively). CONCLUSION: DFR provides similar clinical outcomes compared with LCP for PDFFs. Patients with advanced age may benefit from DFR to allow early weight bearing.

Preliminary prediction of semen quality based on modifiable lifestyle factors by using the XGBoost algorithm.

Introduction: Semen quality has decreased gradually in recent years, and lifestyle changes are among the primary causes for this issue. Thus far, the specific lifestyle factors affecting semen quality remain to be elucidated. Materials and methods: In this study, data on the following factors were collected from 5,109 men examined at our reproductive medicine center: 10 lifestyle factors that potentially affect semen quality (smoking status, alcohol consumption, staying up late, sleeplessness, consumption of pungent food, intensity of sports activity, sedentary lifestyle, working in hot conditions, sauna use in the last 3 months, and exposure to radioactivity); general factors including age, abstinence period, and season of semen examination; and comprehensive semen parameters [semen volume, sperm concentration, progressive and total sperm motility, sperm morphology, and DNA fragmentation index (DFI)]. Then, machine learning with the XGBoost algorithm was applied to establish a primary prediction model by using the collected data. Furthermore, the accuracy of the model was verified via multiple logistic regression following k-fold cross-validation analyses. Results: The results indicated that for semen volume, sperm concentration, progressive and total sperm motility, and DFI, the area under the curve (AUC) values ranged from 0.648 to 0.697, while the AUC for sperm morphology was only 0.506. Among the 13 factors, smoking status was the major factor affecting semen volume, sperm concentration, and progressive and total sperm motility. Age was the most important factor affecting DFI. Logistic combined with cross-validation analysis revealed similar results. Furthermore, it showed that heavy smoking (>20 cigarettes/day) had an overall negative effect on semen volume and sperm concentration and progressive and total sperm motility (OR = 4.69, 6.97, 11.16, and 10.35, respectively), while age of >35 years was associated with increased DFI (OR = 5.47). Conclusion: The preliminary lifestyle-based model developed for semen quality prediction by using the XGBoost algorithm showed potential for clinical application and further optimization with larger training datasets.

Alternative polarization of resident macrophages improves hyperglycemia-associated male infertility.

Recent studies have demonstrated that hyperglycemia induces inflammation in male reproductive system to cause sperm damages and infertility, which may be associated with re-polarization of tissue macrophages from an anti-inflammation M2-like subtype to a pro-inflammation M1-like subtype. However, the underlying mechanisms are not fully determined and a practical approach to interfere with the progression of infertility is lacking. Here, we transduced the testicular macrophages back to the M2-like phenotype with adeno-associated viruses carrying an M2-trigger, Jumonji domain-containing protein D3 (JMJD3), under a macrophage-specific CD68 promoter (CD68p-JMJD3), in streptozotocin-induced diabetic mice. We found that JMJD3-induced M2-polarization of testicular macrophages significantly improved the mating capability of diabetic male mice. The diabetes-induced impairment of the motility of spermatozoa and the decreases in the serum and testicular testosterone levels were both significantly alleviated in CD68p-JMJD3-treated diabetic mice. Thus, our study proposes a practical strategy to treat hyperglycemia-associated infertility.

MRI-based nomogram for differentiation of ovarian fibrothecoma and broad ligament myoma.

Currently, there are no effective approaches for differentiating ovarian fibrothecoma (OF) from broad ligament myoma (BLM). This retrospective study aimed to construct a nomogram prediction model based on MRI to differentiate OF from BLM. The quantitative and qualitative MRI features of 41 OFs and 51 BLMs were compared. Three models were established based on the combination of these features. The ability of the models to differentiate between the two cancers was assessed by ROC analysis. A nomogram based on the best model was constructed for clinical application. The three models showed good performance in differentiating between OF and BLM. The areas under the curve (AUC) of the models based on quantitative and qualitative variables were 0.88 (95% CI: 0.79-0.96) and 0.85 (95% CI: 0.76-0.93), respectively. The combined model designed from the significant variables exhibited the best diagnostic performance with the highest AUC of 0.92 (95% CI: 0.86-0.98). Calibration of the nomogram showed that the predicted probability matched the actual probability well. Analysis of the decision curve demonstrated that the nomogram was clinically useful. Relative T1 value, stone paving sign, enhancement patterns, and ascites were identified as valuable predictors for identifying OF or BLM. The MRI-based nomogram can serve as a preoperative tool to differentiate OF from BLM.

Diagnosis of proteinuria using a random urine protein-creatinine ratio and its correlation with adverse outcomes in pregnancy with preeclampsia characterized by renal damage.

Based on a limited number of studies, a random urine protein-creatinine ratio (uPCR) value of ≥ 0.3 indicates abnormal proteinuria in preeclampsia with renal damage. However, current guidelines do not recommend a reasonable diagnostic threshold of uPCR for severe preeclampsia with renal damage. Furthermore, the correlation between the uPCR value and clinical adverse outcomes remains poorly understood. The aim of the present study was to evaluate the value of uPCR in the diagnosis of significant proteinuria and to assess its correlation with adverse pregnancy outcomes in preeclampsia characterized by renal damage. In all, 1837 women were enrolled in this retrospective cohort study. Eventually, 961 women were enrolled under the exclusion criteria. First, the authors found that uPCR and 24-hour proteinuria showed a significant association (r = 0.901). The optimal threshold of uPCR for diagnosing preeclampsia was 0.295, and for diagnosing severe preeclampsia the cut-off was 0.625. Meanwhile, the adjusted odds ratio per 1 unit increase in ln (uPCR) was 1.679 (95% confidence interval [CI]:1.142-2.469) for severe adverse perinatal outcomes; 1.456 (95% CI: 1.242-1.705) for small for gestational age; 1.380 (95% CI: 1.051-1.811) for severe small for gestational age; 1.672 (95% CI: 1.210-2.310) for very early preterm birth; 1.989 (95% CI 1.726-2.293) for severe hypertension; and 2.279 (95% CI 1.906-2.724) for preterm birth. This study indicated that there was a significant and positive correlation between uPCR and 24-hour urine protein. For neonatal and maternal adverse outcomes, uPCR is an independent predictor of prognosis.

Exosomes derived from hypoxia preconditioned mesenchymal stem cells laden in a silk hydrogel promote cartilage regeneration via the miR-205-5p/PTEN/AKT pathway.

Tissue engineering has promising prospects for cartilage regeneration. However, there remains an urgent need to harvest high quality seed cells. Bone marrow mesenchymal cells (BMSCs), and in particular their exosomes, might promote the function of articular chondrocytes (ACs) via paracrine mechanisms. Furthermore, preconditioned BMSCs could provide an enhanced therapeutic effect. BMSCs naturally exist in a relatively hypoxic environment (1%-5% O2); however, they are usually cultured under higher oxygen concentrations (21% O2). Herein, we hypothesized that hypoxia preconditioned exosomes (H-Exos) could improve the quality of ACs and be more conducive to cartilage repair. In our study, we compared the effects of exosomes derived from BMSCs preconditioned with hypoxia and normoxia (N-Exos) on ACs, demonstrating that H-Exos significantly promoted the proliferation, migration, anabolism and anti-inflammation effects of ACs. Furthermore, we confirmed that hypoxia preconditioning upregulated the expression of miR-205-5p in H-Exos, suggesting that ACs were promoted via the miR-205-5p/PTEN/AKT pathway. Finally, an injectable silk fibroin (SF) hydrogel containing ACs and H-Exos (SF/ACs/H-Exos) was utilized to repair cartilage defects and effectively promote cartilage regeneration in vivo. The application of SF/ACs/H-Exos hydrogel in cartilage regeneration therefore has promising prospects. STATEMENT OF SIGNIFICANCE: Cartilage tissue engineering (CTE) has presented a promising prospect. However, the quality of seed cells is an important factor affecting the repair efficiency. Our study demonstrates for the first time that the exosomes derived from hypoxia preconditioned BMSCs (H-Exos) effectively promote the proliferation, migration and anabolism of chondrocytes and inhibit inflammation through miR-205-5p/PTEN/AKT pathway. Furthermore, we fabricated an injectable silk fibrion (SF) hydrogel to preserve and sustained release H-Exos. A complex composed of SF hydrogel, H-Exos and chondrocytes can effectively promote the regeneration of cartilage defects. Therefore, this study demonstrates that hypoxia pretreatment could optimize the therapeutic effects of BMSCs-derived exosomes, and the combination of exosomes and SF hydrogel could be a promising therapeutic method for cartilage regeneration.

3D Bioprinting of Biomimetic Bilayered Scaffold Consisting of Decellularized Extracellular Matrix and Silk Fibroin for Osteochondral Repair.

Recently, three-dimensional (3D) bioprinting technology is becoming an appealing approach for osteochondral repair. However, it is challenging to develop a bilayered scaffold with anisotropic structural properties to mimic a native osteochondral tissue. Herein, we developed a bioink consisting of decellularized extracellular matrix and silk fibroin to print the bilayered scaffold. The bilayered scaffold mimics the natural osteochondral tissue by controlling the composition, mechanical properties, and growth factor release in each layer of the scaffold. The in vitro results show that each layer of scaffolds had a suitable mechanical strength and degradation rate. Furthermore, the scaffolds encapsulating transforming growth factor-beta (TGF-ß) and bone morphogenetic protein-2 (BMP-2) can act as a controlled release system and promote directed differentiation of bone marrow-derived mesenchymal stem cells. Furthermore, the in vivo experiments suggested that the scaffolds loaded with growth factors promoted osteochondral regeneration in the rabbit knee joint model. Consequently, the biomimetic bilayered scaffold loaded with TGF-ß and BMP-2 would be a promising strategy for osteochondral repair.

Resveratrol Relieves Gouty Arthritis by Promoting Mitophagy to Inhibit Activation of NLRP3 Inflammasomes.

BACKGROUND: Gouty arthritis (GA) is a common inflammatory disease with pain caused by the deposition of monosodium urate (MSU) crystals into joints and surrounding tissues. Resveratrol (Res), derived from grapes and peanuts and the traditional Chinese medicine (TCM) Reynoutria japonica for GA, acts against oxidation and inflammation. The present study aimed to investigate the therapeutic effect and mechanism of Res on GA. METHODS: Arthritis rat models, MSU-induced peritonitis mouse models, and inflammatory models of mouse bone marrow-derived macrophage (BMDM) were used in this study. Enzyme-linked immunosorbent assay (ELISA), JC-1, histopathological, immunofluorescence, flow cytometry, Western blot methods were applied to observe the effects of resveratrol on NLRP3 inflammasomes and mitophagy. RESULTS: Res significantly improves the gait score and synovitis of rats with GA and inhibits the peritoneal inflammation induced by MSU. Res inhibits the MSU-induced activation of NLRP3 inflammasomes by reducing the levels of IL-1ß, IL-18, and Caspase-1 and the pyroptosis of macrophages. In addition, Res raises the level of mitochondrial membrane potential, inhibits the expression of P62 and Pink1, enhances the expressions of LC3B-II, Parkin, and TOMM20, and promotes mitophagy, while mitophagy inhibitors reverse the inhibitory effect of Res on the activation of NLRP3 inflammasomes. CONCLUSION: Res significantly improves GA, and the underlying mechanism might be inhibiting the activation of NLRP3 inflammasomes by triggering the Pink1/Parkin pathway to promote mitophagy.

Preoperative MRI and immunohistochemical examination for the prediction of high-risk endometrial cancer.

BACKGROUND: Magnetic resonance imaging (MRI) and immunohistochemical (IHC) examination provides useful information for the risk stratification of endometrial cancer (EC). However, the use of the combination of MRI and IHC for the prediction of high-risk EC is controversial. The aim of this study was to evaluate the value of preoperative MRI and IHC examination in prediction of patients with high-risk EC. METHODS: This retrospective case-control study was conducted from January 1, 2018 to May 1, 2021 at two hospitals. A primary cohort (n=102) comprised patients with histologically confirmed EC in one hospital between January 1, 2018 and May 31, 2020. An additional external cohort (n=35) comprising patients with histologically confirmed EC in a different hospital from January 1, 2020 to May 1, 2021 was included for validation. Imaging features including tumor size, tumor margin, relative T2 value, tumor signal intensity on diffusion-weighted imaging (DWI), T1-weighted imaging (T1WI), T2-weighted imaging (T2WI) were determined from preoperative MRI images. IHC markers including ER, PR, p53 and Ki67 were determined through IHC staining of preoperative curettage specimen. Patients were divided into high-risk and low-intermediate- risk group based on the final histological results. Differences between categorical and numerical variables were assessed using chi-square test and independent-sample t-test, respectively. Multivariate binary logistic regression analyses were used for construction of the prediction model A fusion prediction model was constructed by combining MRI features and IHC markers. The predictive performance of the model was then validated using the external cohort. RESULTS: Imaging and IHC markers were significantly associated with risk ranks. Model 1 based on MRI features showed an area under the curve (AUC) of 0.822 [95% confidence interval (CI), 0.741-0.903] whereas Model 2 based on IHC markers showed an AUC of 0.894 (95% CI, 0.829-0.960). Notably, model 3 integrating independent MRI and IHC risk factors demonstrated good calibration and high differentiation ability with an AUC of 0.958 (95% CI, 0.923-0.993), and showed good discrimination with an AUC of 0.84 (95% CI, 0.677-0.942) using the external validation set. CONCLUSIONS: This study proposes a comprehensive predictive model comprising MRI and IHC features as a powerful tool for preoperative risk stratification to assist in clinical decision-making for EC patients.

Quantitative control of noise in mammalian gene expression by dynamic histone regulation.

Fluctuation ('noise') in gene expression is critical for mammalian cellular processes. Numerous mechanisms contribute to its origins, yet the mechanisms behind large fluctuations that are induced by single transcriptional activators remain elusive. Here, we probed putative mechanisms by studying the dynamic regulation of transcriptional activator binding, histone regulator inhibitors, chromatin accessibility, and levels of mRNAs and proteins in single cells. Using a light-induced expression system, we showed that the transcriptional activator could form an interplay with dual functional co-activator/histone acetyltransferases CBP/p300. This interplay resulted in substantial heterogeneity in H3K27ac, chromatin accessibility, and transcription. Simultaneous attenuation of CBP/p300 and HDAC4/5 reduced heterogeneity in the expression of endogenous genes, suggesting that this mechanism is universal. We further found that the noise was reduced by pulse-wide modulation of transcriptional activator binding possibly as a result of alternating the epigenetic states. Our findings suggest a mechanism for the modulation of noise in synthetic and endogenous gene expression systems.