Is single-stage bilateral medial opening wedge high tibial osteotomy advisable?

PURPOSE: To validate the safety and clinical results of single-stage bilateral versus unilateral medial opening wedge high tibial osteotomy (HTO). METHODS: A propensity-matched cohort study was performed from March 2020 to March 2021 in our medical center. Data were prospectively collected. Including 34 patients who underwent single-stage bilateral medial opening HTO(SSBHTO), and 68 cases in the unilateral group. Propensity-matched ration was 2:1 based on age, sex, and body mass index using R software. Comparisons of the length of hospital stay, operative time, blood loss, postoperative adverse events, 90-day readmission rate, conversion to TKA rate, self-reported VAS and WOMAC scores were made to investigate the safety and clinical results of bilateral HTO. RESULTS: The mean length of hospital stay was 7.36 ± 2.23 days for SSBHTO and 7 days (IQR, 3 days; range, 4 to 23 days) for the unilateral group (P = 0.219). The mean operative time was 144 ± 47 min for bilateral HTO and 105(37.5) mins for a unilateral OWHTO (P < 0.001). The mean blood loss was 150(100) ml for SSBHTO and 100(50) ml for unilateral OWHTO (P < 0.001). There were no significant difference of the adverse events and 90-day readmission rate between two groups. No failed HTO or conversion to knee arthroplasty were observed at the end of follow-up. VAS, pain, stiffness, and functional scores of the WOMAC scale were essentially comparable of two groups one year after surgery (P > 0.05). CONCLUSIONS: A single-stage bilateral medial opening wedge high tibial osteotomy is advisable for patients with knee osteoarthritis. Patients benefit from avoiding secondary anesthesia, postoperative complications, and substantial cost savings. LEVEL OF EVIDENCE: Therapeutic Level III.

Antitumor Effect and Immunomodulatory Mechanism of "Oncolytic Extracellular Vesicles".

Enhancing the antitumor immune response and targeting ability of oncolytic viruses will improve the effect of tumor immunotherapy. Through infecting neural stem cells (NSCs) with a capsid dual-modified oncolytic adenovirus (CRAd), we obtained and characterized the "oncolytic extracellular vesicles" (CRAdEV) with improved targeted infection and tumor killing activity compared with CRAd. Both ex vivo and in vivo studies revealed that CRAdEV activated innate immune cells and importantly enhanced the immunomodulatory effect compared to CRAd. We found that CRAdEV effectively increased the number of DCs and activated CD4+ and CD8+ T cells, significantly increased the number and activation of B cells, and produced higher levels of tumor-specific antibodies, thus eliciting enhanced antitumor activity compared with CRAd in a B16 xenograft immunocompetent mice model. This study provides a novel approach to oncolytic adenovirus modification and demonstrates the potential of "oncolytic extracellular vesicles" in antitumor immunotherapy.

Dissecting causal relationships between primary biliary cholangitis and extrahepatic autoimmune diseases based on Mendelian randomization.

As an autoimmune disease, up to 73% of patients with primary biliary cholangitis (PBC) have a combination of extrahepatic autoimmune diseases (EHAIDs); however, the causal relationship between PBC and EHAIDs is unclear. The genome-wide association analyses provided 14 GWAS data for PBC and EHAIDs, and bidirectional, two-sample MR analyses were performed to examine the relationship between PBC and EHAIDs. The analysis using MR provides a strong and meaningful estimation of the bidirectional correlation between PBC and 7 EHAIDs: rheumatoid arthritis, systemic lupus erythematosus, Sjögren's syndrome, systemic sclerosis, autoimmune hypothyroidism, inflammatory bowel disease and ulcerative colitis of its types. In addition, PBC increases the risk of autoimmune thyroid diseases such as autoimmune hyperthyroidism and Graves' disease, as well as multiple sclerosis and psoriasis. Additionally, PBC is identified as a risk factor for Crohn's disease and Celiac disease. Based on genetic evidence, there may be connections between PBC and specific EHAIDs: not all coexisting EHAIDs induce PBC, and vice versa. This underscores the significance of prioritizing PBC in clinical practice. Additionally, if any liver function abnormalities are observed during treatment or with EHAIDs, it is crucial to consider the possibility of comorbid PBC.

Increased symptoms of stiffness after opening-wedge high tibial osteotomy are associated with worse postoperative knee function outcomes and lower patient satisfaction rate.

BACKGROUND: Symptoms of knee stiffness after open wedge high tibial osteotomy (OW-HTO) can significantly affect surgical effectiveness, but no studies have reported risk factors for knee stiffness after OW-HTO. METHODS: Patients treated with OW-HTO for the first time between 2018 and 2021 were included. Data were collected on patient demographics, Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC), Short Form (SF) 12 scores, hip-knee-ankle angle (HKA) and patient satisfaction before and after surgery. Patients with worse WOMAC stiffness scores at 1 year were defined as the 'increased stiffness' group and the other cohort as the 'non-stiffness' group. The primary outcome of the study was to compare postoperative knee function scores (WOMAC and SF-12), HKA and patient satisfaction rate between the two groups. The secondary outcome was the use of logistic regression to analyze independent predictors of increased postoperative stiffness symptoms. RESULTS: At 1 year postoperatively, 95 (11.3%) patients had a significant increase in stiffness. Patients had significantly (p < .001) less improvement in pain, function, and total WOMAC scores, and SF-12 score than those in the non-stiffness group (n = 745). However, the differences in WOMAC and SF-12 scores in increased stiffness group at 1 year post-operatively were statistically significant (p < .001) compared to the non-stiffness group. There was no statistically significant difference in HKA in the increased stiffness group (172.9° ± 2.3°) compared to non-stiffness group (173.4° ± 2.6°) at 1 year postoperatively (p = .068). Patient satisfaction was significantly lower in the increased stiffness group (p < .001). Logistic regression analysis showed that diabetes (odds ratio (OR) 1.809, p = .034) and preoperative WOMAC stiffness score of 44 or less (OR 4.255 p < .001) were predictors of increased stiffness. CONCLUSIONS: Patients with increased stiffness after OW-HTO had worse functional outcomes and lower patient satisfaction rates and patients at risk of being in this group should be informed pre-operatively.

Risk Factors and Dynamic Nomogram Development for Surgical Site Infection Following Open Wedge High Tibial Osteotomy for Varus Knee Osteoarthritis: A Retrospective Cohort Study.

Background: As the worldwide population ages, the population receiving open wedge high tibial osteotomy (OWHTO) is growing, and surgical site infection (SSI) is a rare but fatal surgical complication. This study aimed to identify risk factors independently associated with SSI following OWHTO and develop a predictive nomogram. Methods: Clinical data of patients who received OWHTO and followed up for more than 12 months in our hospital were retrospectively reviewed. Multivariable logistic regression was performed to determine independent risk factors for SSI and to construct predictive nomograms. The study further illustrated the predictive performance of the model by using the receiver operating characteristic (ROC) curve, calibration curve, and decision curve analysis (DCA). Results: A total of 1294 eligible patients were included in the study. Multivariate analysis revealed tobacco consumption (OR=3.44, p=0.010), osteotomy size ≥12 mm (OR=3.3, p=0.015), the use of allogeneic bone or artificial bone graft substitutes (allogeneic bone vs none, OR=4.08, p=0.037; artificial bone vs none, OR=5.16, p=0.047), Kellgren-Lawrence (K-L) grade IV (OR=2.5, p=0.046), systemic immune-inflammation index (SII) >423.62 (OR=6.2, p<0.001), high-sensitivity C-reactive protein (HCRP) >2.6 mg/L (OR=2.42, p=0.044), and a higher level of fasting blood glucose (FBG) (OR=1.32, p=0.022) were the independent predictors of SSI. The cutoff score of the model was 148, with a sensitivity of 76.0% and specificity of 81.0%. The concordance index (C-index) and Brier score of the nomogram were 0.856 and 0.017, and the corrected values after 1000 bootstrapping validations were 0.820 and 0.018, respectively. Furthermore, the ROC curve, calibration curve, and DCA exhibited excellent predictive accuracy and clinical applicability of the model. Conclusion: This study developed a dynamic nomogram based on seven predictors, which allowed surgeons to individualize risk stratification of patients and intervene promptly to reduce SSI rates.

Cell Membrane-Coated Oncolytic Adenovirus for Targeted Treatment of Glioblastoma.

An oncolytic virus is a promising strategy for glioblastoma (GBM) therapy. However, there are still some challenges such as the blood-brain barrier (BBB) and preexisting immunity for targeted treatment of GBM with an oncolytic virus. In this study, two kinds of cell membrane-coated oncolytic adenoviruses (NCM-Ad and GCM-Ad) were prepared using neural stem cells (NSCs) and GBM cells as sources of membranes, respectively, and were shown to improve the targeted infectivity on GBM cells and avoid the immune clearance of preexisting neutralizing antibodies in vitro and in vivo. Specifically, NCM-Ad showed a strong ability to cross the BBB and target tumor cells in vivo. To improve the cytotoxicity to GBM, a capsid dual-modified oncolytic adenovirus (A4/k37) was also encapsulated, and NCM-A4/k37 showed outstanding tumor targeting and inhibition capacity in an orthotopic xenograft tumor model of GBM upon intravenous administration. This study provides a promising oncolytic virus-based targeted therapeutic strategy for glioma.

Dose-related immunomodulatory effects of recombinant TRAIL in the tumor immune microenvironment.

BACKGROUND: In addition to specifically inducing tumor cell apoptosis, recombinant tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) has also been reported to influence the cancer immune microenvironment; however, its underlying effects and mechanisms remain unclear. Investigating the immunomodulatory effects and mechanisms of recombinant TRAIL in the tumor microenvironment (TME) may provide an important perspective and facilitate the exploration of novel TRAIL strategies for tumor therapy. METHODS: Immunocompetent mice with different tumors were treated with three doses of recombinant TRAIL, and then the tumors were collected for immunological detection and mechanistic investigation. Methodological approaches include flow cytometry analysis and single-cell sequencing. RESULTS: In an immunocompetent mouse model, recombinant soluble mouse TRAIL (smTRAIL) had dose-related immunomodulatory effects. The optimal dose of smTRAIL (2 mg/kg) activated innate immune cells and CD8+ T cells, whereas higher doses of smTRAIL (8 mg/kg) promoted the formation of a tumor-promoting immune microenvironment to counteract the apoptotic effects on tumor cells. The higher doses of smTRAIL treatment promoted M2-like macrophage recruitment and polarization and increased the production of protumor inflammatory cytokines, such as IL-10, which deepened the suppression of natural killer (NK) cells and CD8+ T cells in the tumor microenvironment. By constructing an HU-HSC-NPG.GM3 humanized immune system mouse model, we further verified the immunomodulatory effects induced by recombinant soluble human TRAIL (shTRAIL) and found that combinational administration of shTRAIL and trabectedin, a macrophage-targeting drug, could remodel the tumor immune microenvironment, further enhance antitumor immunity, and strikingly improve antitumor effects. CONCLUSION: Our results highlight the immunomodulatory role of recombinant TRAIL and suggest promising therapeutic strategies for clinical application.

Proximal tibial osteotomy with absorbable spacer combined with fibular osteotomy has similar clinical outcomes to high tibial osteotomy in the treatment of knee osteoarthritis.

PURPOSE: Proximal tibia osteotomy with absorbable spacer combined with fibular osteotomy (TPOASI) is an emerging surgical technique for treating knee osteoarthritis (KOA); however, the efficacy of this procedure remains unknown. We hypothesize that TPOASI can achieve similar clinical outcomes to opening-wedge high tibial osteotomy (OW-HTO). The objective of this study is to compare the clinical results between these two procedures. METHODS: Patients who underwent TPOASI or OW-HTO from July 2016 to September 2020 were included. The following outcome parameters were determined before and after the surgery: the Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC), the Knee Injury and Osteoarthritis Outcome Score (KOOS), the visual analogue scale of pain, the Intermittent and Persistent Osteoarthritis Pain Scale, femorotibial angle, and post-operative complications. RESULTS: In total, 209 cases were analyzed (102 in TPOASI group; 107 in OW-HTO group) with 3.1 years average follow-up. Both procedures achieved significant improvement in KOOS (62.0 to 24.4 in the TPOASI and 62.8 to 26.2 in the OW-HTO group, p < 0.001) and WOMAC score (68.9 to 24.1 in the TPOASI versus 69.9 to 26.1 in the OW-HTO group, p < 0.001). There were no significant differences in complications or femorotibial angle between the two groups but the only significant difference in the outcome parameters was the WOMAC stiffness score (19.6 in the TPOASI versus 26.5 in the OW-HTO group). CONCLUSION: TPOASI achieves comparable results to OW-HTO in terms of clinical scores, radiographic results, and complications, but has the advantage of avoiding internal fixation removal.

Development and validation of a nomogram for predicting the risk of immediate postoperative deep vein thrombosis after open wedge high tibial osteotomy.

PURPOSE: This study aimed to identify independent risk factors for immediate postoperative deep vein thrombosis (DVT) in patients with open wedge high tibial osteotomy (OWHTO) and to develop and validate a predictive nomogram. METHODS: Patients who underwent OWHTO for knee osteoarthritis (KOA) from June 2017 to December 2021 were retrospectively analyzed. Baseline data and laboratory test results were collected, and the occurrence of DVT in the immediate postoperative period was regarded as the study outcome event. Multivariable logistic regression identified independent risk factors associated with a higher incidence of immediate postoperative DVT. The predictive nomogram was constructed based on the analysis results. The stability of the model was further assessed in this study using patients from January to September 2022 as an external validation set. RESULTS: 741 patients were enrolled in the study, of which 547 were used in the training cohort and the other 194 for the validation cohort. Multivariate analysis revealed a higher Kellgren-Lawrence (K-L) grade (III vs. I-II OR 3.09, 95% CI 0.93-10.23. IV vs. I-II OR 5.23, 95% CI 1.27-21.48.), platelet to hemoglobin ratio (PHR) > 2.25 (OR 6.10, 95% CI 2.43-15.33), Low levels of albumin (ALB) (OR 0.79, 95% CI 0.70-0.90), LDL-C > 3.40 (OR 3.06, 95% CI 1.22-7.65), D-dimer > 1.26 (OR 2.83, 95% CI 1.16-6.87) and BMI ≥ 28 (OR 2.57, 95% CI 1.02-6.50) were the independent risk factors of immediate postoperative DVT. The concordance index (C-index) and Brier score of the nomogram were 0.832 and 0.036 in the training set, and the corrected values after internal validation were 0.795 and 0.038, respectively. The receiver-operating characteristic (ROC) curve, the calibration curve, the Hosmer-Lemeshow test, and the decision curve analysis (DCA) performed well in both the training and validation cohorts. CONCLUSION: This study developed a personalized predictive nomogram with six predictors, which allows surgeons to stratify risk and recommended immediate ultrasound scans for patients with any of these factors. LEVEL OF EVIDENCE: III.

By-Products of Fruit and Vegetables: Antioxidant Properties of Extractable and Non-Extractable Phenolic Compounds.

Non-extractable phenolic compounds (NEPs), or bound phenolic compounds, represent a crucial component of polyphenols. They are an essential fraction that remains in the residual matrix after the extraction of extractable phenolic compounds (EPs), making them a valuable resource for numerous applications. These compounds encompass a diverse range of phenolic compounds, ranging from low molecular weight phenolic to high polymeric polyphenols attached to other macro molecules, e.g., cell walls and proteins. Their status as natural, green antioxidants have been well established, with numerous studies showcasing their anti-inflammatory, anti-aging, anti-cancer, and hypoglycemic activities. These properties make them a highly desirable alternative to synthetic antioxidants. Fruit and vegetable (F&Veg) wastes, e.g., peels, pomace, and seeds, generated during the harvest, transport, and processing of F&Vegs, are abundant in NEPs and EPs. This review delves into the various types, contents, structures, and antioxidant activities of NEPs and EPs in F&Veg wastes. The relationship between the structure of these compounds and their antioxidant activity is explored in detail, highlighting the importance of structure-activity relationships in the field of natural antioxidants. Their potential applications ranging from functional food and beverage products to nutraceutical and cosmetic products. A glimpse into their bright future as a valuable resource for a greener, healthier, and more sustainable future, and calling for researchers, industrialists, and policymakers to explore their full potential, are elaborated.

Vertebral bone quality score provides preoperative bone density assessment for patients undergoing lumbar spine surgery: a retrospective study.

OBJECTIVE: The novel MRI-based vertebral bone quality (VBQ) score has been described as an opportunistic screening tool for osteoporosis, but the stability and practical value of this score deserve further investigation. The purpose of this study was to assess whether preoperative VBQ scores could assist in identifying reduced bone mineral density (BMD) or osteoporosis and evaluating the consistency between MRI systems with different field strengths. METHODS: The VBQ scores of the patients who underwent surgery for lumbar disc herniation and the single-level VBQ scores of each L1-4 vertebral body were measured and calculated with preoperative lumbar MRI noncontrast T1-weighted phases. The VBQ scores were evaluated for correlation analysis using dual-energy x-ray absorptiometry (DEXA) T-scores. The receiver operating characteristic (ROC) curve was used to evaluate the ability of the VBQ scores to identify patients with reduced BMD and with osteoporosis. Differences in CSF measurements at different levels of L1-4 were compared. Twenty-four patients who had been examined using another MRI machine were used as controls to test the interdevice agreement of the VBQ scores. RESULTS: The study included 100 patients with mean VBQ scores of 2.81 ± 0.28 (normal BMD), 3.06 ± 0.36 (osteopenia), and 3.43 ± 0.37 (osteoporosis). VBQ scores differed significantly between BMD subgroups (p < 0.001). The Pearson correlation coefficient showed a moderate negative linear correlation between novel VBQ scores and the lowest DEXA T-scores (r = -0.524). ROC analysis showed good discrimination of VBQ scores in patients with reduced BMD (area under the curve [AUC] 0.793) and with osteoporosis (AUC 0.810). The diagnostic thresholds of reduced BMD and osteoporosis according to the maximum Youden index were 3.06 (sensitivity 0.636, specificity 0.870, positive predictive value [PPV] 0.942, negative predictive value [NPV] 0.417) and 3.05 (sensitivity 0.875, specificity 0.618, PPV 0.519, NPV 0.913), respectively. CSF measurements at the L2, L3, and L4 levels were essentially identical and did not significantly affect the final VBQ scores (p > 0.05), whereas CSF measurements at the L1 level were found to be heterogeneous (p < 0.001). No significant differences were found in VBQ scores between the same brand of MRI machines at different field strengths (1.5 and 3.0 T, p = 0.107). CONCLUSIONS: The new VBQ score provides an additional screening opportunity for preoperative BMD assessment. A VBQ score < 3.05 essentially excludes osteoporosis, whereas a VBQ score ≥ 3.05 (especially ≥ 3.06) suggests the need for further examination. The VBQ score is comparable between different MRI systems.

Activation of Wnt/ß-Catenin Signaling Involves 660 nm Laser Radiation on Epithelium and Modulates Lipid Metabolism.

Research has proven that light treatment, specifically red light radiation, can provide more clinical benefits to human health. Our investigation was firstly conducted to characterize the tissue morphology of mouse breast post 660 nm laser radiation with low power and long-term exposure. RNA sequencing results revealed that light exposure with a higher intervention dosage could cause a number of differentially expressed genes compared with a low intervention dosage. Gene ontology analysis, protein-protein interaction network analysis, and gene set enrichment analysis results suggested that 660 nm light exposure can activate more transcription-related pathways in HC11 breast epithelial cells, and these pathways may involve modulating critical gene expression. To consider the critical role of the Wnt/T-catenin pathway in light-induced modulation, we hypothesized that this pathway might play a major role in response to 660 nm light exposure. To validate our hypothesis, we conducted qRT-PCR, immunofluorescence staining, and Western blot assays, and relative results corroborated that laser radiation could promote expression levels of ß-catenin and relative phosphorylation. Significant changes in metabolites and pathway analysis revealed that 660 nm laser could affect nucleotide metabolism by regulating purine metabolism. These findings suggest that the Wnt/ß-catenin pathway may be the major sensor for 660 nm laser radiation, and it may be helpful to rescue drawbacks or side effects of 660 nm light exposure through relative interventional agents.

Endogenous Follistatin-like 1 guarantees the immunomodulatory properties of mesenchymal stem cells during liver fibrotic therapy.

BACKGROUND: Mesenchymal stem cell (MSC) therapy has been shown to be a promising option for liver fibrosis treatment. However, critical factors affecting the efficacy of MSC therapy for liver fibrosis remain unknown. Follistatin-like 1 (FSTL1), a TGF-ß-induced matricellular protein, is documented as an intrinsic regulator of proliferation and differentiation in MSCs. In the present study, we characterized the potential role of FSTL1 in MSC-based anti-fibrotic therapy and further elucidated the mechanisms underlying its action. METHODS: Human umbilical cord-derived MSCs were characterized by flow cytometry. FSTL1low MSCs were achieved by FSTL1 siRNA. Migration capacity was evaluated by wound-healing and transwell assay. A murine liver fibrotic model was created by carbon tetrachloride (CCl4) injection, while control MSCs or FSTL1low MSC were transplanted via intravenous injection 12 weeks post CCl4 injection. Histopathology, liver function, fibrosis degree, and inflammation were analysed thereafter. Inflammatory cell infiltration was evaluated by flow cytometry after hepatic nonparenchymal cell isolation. An MSC-macrophage co-culture system was constructed to further confirm the role of FSTL1 in the immunosuppressive capacity of MSCs. RNA sequencing was used to screen target genes of FSTL1. RESULTS: FSTL1low MSCs had comparable gene expression for surface markers to wildtype but limited differentiation and migration capacity. FSTL1low MSCs failed to alleviate CCl4-induced hepatic fibrosis in a mouse model. Our data indicated that FSTL1 is essential for the immunosuppressive action of MSCs on inflammatory macrophages during liver fibrotic therapy. FSTL1 silencing attenuated this capacity by inhibiting the downstream JAK/STAT1/IDO pathway. CONCLUSIONS: Our data suggest that FSTL1 facilitates the immunosuppression of MSCs on macrophages and that guarantee the anti-fibrotic effect of MSCs in liver fibrosis.

A novel immune checkpoint-related gene signature for hepatocellular carcinoma to predict clinical outcomes and therapeutic response.

Immune checkpoint genes (ICGs) have recently been proven to perform instrumental functions in the maintenance of immune homeostasis and represent a promising therapeutic strategy; however, their expression patterns and prognostic values are not fully elucidated in hepatocellular carcinoma (HCC). In this investigation, we focused on establishing and validating a prognostic gene signature to facilitate decision-making in clinical practice. Clinical information, as well as transcriptome data, was obtained from the Cancer Genome Atlas (TCGA) and International Cancer Genome Consortium (ICGC) database. Univariate Cox regression and least absolute shrinkage and selection operator (LASSO) Cox method were employed to build a multi-gene signature in the TCGA database, while the ICGC database was used for validation. Subsequently, utilizing the six-gene signature, we were able to categorize patients into high- and low-risk groups. In two cohorts, survival analysis findings revealed a dismal outlook for the high-risk group. The receiver operating characteristic curves were utilized to estimate the gene signature's prediction ability. Moreover, correlation analysis showed high-risk group was linked to advanced pathological stage, infiltration of immune cells and therapeutic response. In summary, this unique gene profile might serve not only as a useful prognostic indicator but also as a marker of therapy responsiveness in HCC.

Bioinformatics Analysis Combined With Experiments Predicts PUDP as a Potential Prognostic Biomarker for Hepatocellular Carcinoma Through Its Interaction With Tumor Microenvironment.

Hepatocellular carcinoma (HCC) is one of the deadliest tumors in the world and is notorious for poor prognosis. There is mounting evidence that pseudouridine performs key functions in the initiation and progression of several cancers. A previous study demonstrated that Pseudouridine 5'-phosphatase (PUDP) may be a novel prognostic biomarker in colorectal cancer. However, in the past, we have paid little attention to PUDP and we are still not clear about its function and role in cancer. In this study, a pan-cancer analysis of PUDP expression and prognosis was performed firstly using The Cancer Genome Atlas (TCGA) and Genotype-Tissue Expression (GTEx) data and we found that PUDP may be a potential oncogene for HCC. Then the most potential upstream microRNA contributing to PUDP was identified as let-7c-5p through expression analysis, correlation analysis, and survival analysis. Subsequently, the result of single cell RNA sequencing (scRNA-seq) demonstrated that PUDP was significantly highly expressed on malignant cells. In addition, there are significantly positive correlations between PUDP and tumor immune cell infiltration, biomarkers of immune cells, and immune checkpoint expression, especially with tumor-promoting immune cells such as T cell regulatory (Treg), Myeloid-derived suppressor cell (MDSC), cancer-associated fibroblast (CAF). Moreover, we found the methylation level of three loci was positively correlated with PUDP expression and four loci were negatively correlated. 15 pairs of HCC and normal adjacent tissues from HCC patients who were treated at our center were used to verify the results of the bioinformatics analysis and the results of experiments are similar to the bioinformatics analysis. Our study demonstrated that HCC patients with high PUDP expression are less likely to benefit from immunotherapy, and in addition, we explored the relationship between PUDP and anticancer drugs. Finally, we explored the clinical relevance of PUDP, identified PUDP as an independent risk factor for HCC patients and constructed a prognostic model, used International Cancer Genome Consortium (ICGC) data to do external validation. Collectively, our study demonstrated that high expression of PUDP suggested a poor prognosis and low response to immunotherapy, providing new insight into the treatment and prognosis of HCC.

Oncolytic adenovirus-mediated intratumoral expression of TRAIL and CD40L enhances immunotherapy by modulating the tumor microenvironment in immunocompetent mouse models.

The immune status of the tumor microenvironment is a key indicator determining the antitumor effect of immunotherapy. Oncolytic viruses directly target tumor cells or indirectly modulate the tumor microenvironment (TME) especially when properly armed. It was previously demonstrated that conditionally replicating adenovirus serotype 5 (CRAd5) encoding tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) had outstanding antitumor effects in different human cancer cells xenograft models; however, its antitumor immune mechanism has not been evaluated in immunocompetent preclinical mouse models. We first explored the antitumor activity of CRAd5-TRAIL in several murine tumor models and found that the expression of TRAIL induced increases or activation in tumor-infiltrating T cells. To further improve the antitumor effects, mouse CD40 ligand (mCD40L) as an immune activator expressed by recombinant Ad5 vector was firstly used in combination with CRAd5-TRAIL for tumor immunotherapy. Both in vitro and in vivo studies demonstrated that mCD40L effectively activated dendritic cells (DCs), B cells, and tumor-infiltrating T cells, and also promoted tumor cell apoptosis by increasing the expression of TRAIL receptors, thereby significantly enhancing the antitumor activity of oncolytic adenoviruses in CT26 and B16 tumor-bearing models. Although affected by the restriction of oncolytic adenovirus replication in mouse cells, the combination treatment failed to completely eliminate tumor cells, our research still provided a promising strategy for oncolytic adenovirus-mediated solid tumor immunotherapy.

Identification and validation a costimulatory molecule gene signature to predict the prognosis and immunotherapy response for hepatocellular carcinoma.

BACKGROUND: Hepatocellular carcinoma (HCC) is one of the most common malignancies worldwide. Costimulatory molecules have been proven to be the foundation of immunotherapy. However, the potential roles of costimulatory molecule genes (CMGs) in HCC remain unclear. Our study is aimed to develop a costimulatory molecule-related gene signature that could evaluate the prognosis of HCC patients. METHODS: Based on The Cancer Gene Atlas (TCGA) database, univariate Cox regression analysis was applied in CMGs to identify prognosis-related CMGs. Consensus clustering analysis was performed to stratify HCC patients into different subtypes and compared them in OS. Subsequently, the LASSO Cox regression analysis was performed to construct the CMGs-related prognostic signature and Kaplan-Meier survival curves as well as ROC curve were used to validate the predictive capability. Then we explored the correlations of the risk signature with tumor-infiltrating immune cells, tumor mutation burden (TMB) and response to immunotherapy. The expression levels of prognosis-related CMGs were validated based on qRT-PCR and Human Protein Atlas (HPA) databases. RESULTS: All HCC patients were classified into two clusters based on 11 CMGs with prognosis values and cluster 2 correlated with a poorer prognosis. Next, a prognostic signature of six CMGs was constructed, which was an independent risk factor for HCC patients. Patients with low-risk score were associated with better prognosis. The correlation analysis showed that the risk signature could predict the infiltration of immune cells and immune status of the immune microenvironment in HCC. The qRT-PCR and immunohistochemical results indicated six CMGs with differential expression in HCC tissues and normal tissues. CONCLUSION: In conclusion, our CMGs-related risk signature could be used as a prediction tool in survival assessment and immunotherapy for HCC patients.

Immunoglobulin M: A Neglected Serum Biomarker in Treatment-Naive Primary Biliary Cholangitis With Normal Alkaline Phosphatase.

The diagnosis of primary biliary cholangitis (PBC) in patients with seropositive anti-mitochondrial antibody (AMA) but normal alkaline phosphatase (ALP) depends on a liver biopsy. We aimed to reveal potential serum biomarkers that could suggest the necessity of a liver biopsy in such patients. Retrospective analysis was performed. Subjects who were treatment naive with seropositive AMA but normal ALP and who underwent at least one liver biopsy between 2008 and 2020 were included in this study. Histologic biopsies were evaluated by two experienced pathologists blinded to the serum tests. A total of 115 patients who were treatment naive were included in this study. Of these, 77 patients (67%) exhibited histologic PBC features and nonspecific histologic features were found in the remaining 38 (33%) patients. Multivariate analysis suggested that baseline serum immunoglobulin M (IgM) >0.773 × upper limit of normal (ULN) (P < 0.001) and age >42 years (P = 0.002) were associated with the diagnosis of PBC through liver biopsies. A significant decrease in the median levels of gamma-glutamyl transpeptidase (GGT) and IgM was found in 54 patients with PBC who received ursodeoxycholic acid (UDCA). Conclusion: For patients who were treatment naive with seropositive AMA but normal ALP, baseline serum IgM >0.773 × ULN and age >42 years were the factors that strongly suggested a diagnosis of PBC. In these patients receiving UDCA, a dynamic monitoring of GGT and IgM might be helpful in evaluating therapeutic responses.

Systematic Construction and Validation of a Prognostic Model for Hepatocellular Carcinoma Based on Immune-Related Genes.

Hepatocellular carcinoma (HCC), a highly aggressive tumor, has high incidence and mortality rates. Recently, immunotherapies have been shown to be a promising treatment in HCC. The results of either the CheckMate-040 or IMbrave 150 trials demonstrate the importance of immunotherapy in the systemic treatment of liver cancer. Thus, in this study, we tried to establish a reliable prognostic model for liver cancer based on immune-related genes (IRGs) and to provide a new insight for immunotherapy of HCC. In this study, we used four datasets that incorporated 851 HCC samples, including 340 samples with complete clinical information from the cancer genome atlas (TCGA) database, to establish an effective model for predicting the prognosis of HCC patients based on the differential expression of IRGs and validated the prognostic model using the data from International Cancer Genome Consortium (ICGC). The top 6 characteristic IRGs identified by protein-protein interaction (PPI) network analysis, MMP9, FOS, CAT, ESR1, ANGPTL3, and KLKB1, were selected for further study. In addition, we assessed the correlations of the six characteristic IRGs with the tumor immune microenvironment, clinical stage, and sensitivity to anti-cancer drugs. We also explored whether the differential expression of the characteristic IRGs was specific to HCC or present in pan-cancer. The expression levels of the six characteristic IRGs were significantly different between most tumor tissues and adjacent normal tissues. In addition, these characteristic IRGs showed a strong association with immune cell infiltration in HCC patients. We found that MMP9 and ESR1 were independent prognostic factors for HCC, while CAT, ESR1, and KLKB1 were associated with the clinical stage. We collected HCC paraffin sections from 24 patients from Xijing hospital to identify the differential expression of the five genes (MMP9, ESR1, CAT, FOS, and KLKB1). Finally, the results of decision curve analysis (DCA) and nomogram revealed that our models provided a prognostic benefit for most HCC patients and the predicted overall survival (OS) was consistent with the actual OS. In conclusion, we systemically constructed a novel prognostic model that provides new insights into HCC.

Rejuvenated ageing mesenchymal stem cells by stepwise preconditioning ameliorates surgery-induced osteoarthritis in rabbits.

AIMS: Ageing-related incompetence becomes a major hurdle for the clinical translation of adult stem cells in the treatment of osteoarthritis (OA). This study aims to investigate the effect of stepwise preconditioning on cellular behaviours in human mesenchymal stem cells (hMSCs) from ageing patients, and to verify their therapeutic effect in an OA animal model. METHODS: Mesenchymal stem cells (MSCs) were isolated from ageing patients and preconditioned with chondrogenic differentiation medium, followed by normal growth medium. Cellular assays including Bromodeoxyuridine / 5-bromo-2'-deoxyuridine (BrdU), quantitative polymerase chain reaction (q-PCR), ß-Gal, Rosette forming, and histological staining were compared in the manipulated human mesenchymal stem cells (hM-MSCs) and their controls. The anterior cruciate ligament transection (ACLT) rabbit models were locally injected with two millions, four millions, or eight millions of hM-MSCs or phosphate-buffered saline (PBS). Osteoarthritis Research Society International (OARSI) scoring was performed to measure the pathological changes in the affected joints after staining. Micro-CT analysis was conducted to determine the microstructural changes in subchondral bone. RESULTS: Stepwise preconditioning approach significantly enhanced the proliferation and chondrogenic potential of ageing hMSCs at early passage. Interestingly, remarkably lower immunogenicity and senescence was also found in hM-MSCs. Data from animal studies showed cartilage damage was retarded and subchondral bone remodelling was prevented by the treatment of preconditioned MSCs. The therapeutic effect depended on the number of cells applied to animals, with the best effect observed when treated with eight millions of hM-MSCs. CONCLUSION: This study demonstrated a reliable and feasible stepwise preconditioning strategy to improve the safety and efficacy of ageing MSCs for the prevention of OA development. Cite this article: Bone Joint Res 2021;10(1):10-21.