An extracellular vesicle microRNA-initiated 3D DNAzyme motor for colorectal cancer diagnosis.

MicroRNA is regarded as a significant biomarker for cancer diagnosis, disease process evaluation and therapeutic guidance, and dual-parameter measurement may contribute to a more accurate and realistic assessment. To meet the urgent need for simultaneous detection of multiple biomarkers, we combined three-dimensional DNAzyme motors with single molecule imaging technique to construct a convenient, intuitive, and sensitive approach for the simultaneous detection of dual miRNAs in the free state or in extracellular vesicles. Quantification of target miRNAs can be realized through the detection of amplified fluorescence signals generated by the target miRNA-initiated cleavage of fluorescent substrate strands by the DNAzyme motors. The practicability was systematically validated with microRNA-21-5p and microRNA-10b-5p as targets, acquiring a satisfactory sensitivity sufficient to detect low abundance targets at 0.5 or 1 pM to 100 pM. Besides, the extracellular vesicular miRNAs can be conveniently detected without extraction. The clinical applicability was verified with a series of extracellular vesicles from clinical samples, which exhibited good distinguishability between colorectal cancer patients and healthy donors. In addition to the advantages of good specificity and high sensitivity, the system has potential to be easily adapted by minor alteration of the DNA sequences and fluorophore sets for detection of multiple miRNAs and even other types of biomarkers such as proteins. Therefore, it shows promise to be widely applied in various fields such as early diagnosis of cancer and its prognostic assessment.

Efficacy analysis of clinical serological indicators in the diagnosis of postoperative periprosthetic joint infection in patients with rheumatoid arthritis or osteoarthritis.

PURPOSE: To investigate the incidence of periprosthetic joint infection (PJI) in patients with rheumatoid arthritis (RA) or osteoarthritis (OA) after primary joint arthroplasty; to analyze the optimal cut-off values of clinical serum markers C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), and D-dimer for the diagnosis of PJI in RA patients; and to explore their diagnostic efficacy and clinical significance. METHODS: Clinical data of 15,702 patients with RA (578) or OA (15,124) who underwent total joint arthroplasty from 2013 to 2021 were retrospectively analyzed. Serum CRP, ESR, and D-dimer were recorded for each patient, and subject characteristic curves were used to determine the optimal threshold values of CRP, ESR, and D-dimer for RA-PJI and OA-PJI and to compare the areas under the curves to assess the diagnostic efficacy of the optimal threshold values of serologic indices for RA-PJI. RESULTS: The five year incidence of PJI was 6.92% in RA patients and 0.67% in OA patients. The optimal thresholds of CRP, ESR, and D-dimer for the diagnosis of RA-PJI were respectively 13.85 mg/L, 33.02 mm/h, and 796.50 ng/mL. The sensitivities of the optimal thresholds were respectively 67.6%, 62.2%, and 56.8%, and the specificities were 74.7%, 60.4%, and 74.4%. CONCLUSION: RA patients have a higher incidence of PJI than OA patients. The optimal thresholds for CRP, ESR, and d-dimer for the diagnosis of PJI were higher in RA patients than in OA patients, but the sensitivity and specificity of the diagnosis were not as good as in OA patients.

Reversible and Highly Ordered Biointerfaces for Efficient Capture and Nondestructive Release of Circulating Tumor Cells.

The engineering strategy of artificial biointerfaces is vital for governing their performances in bioanalysis and diagnosis. Highly ordered arrangement of affinity ligands on the interface surface facilitates efficient interaction with target molecules, whereas biointerfaces aimed at drug delivery or rare cell isolation require sophisticated stimuli-response mechanisms. However, it is still challenging to facilely fabricate biointerfaces possessing the two features. Herein, we endow a biointerface with both reversibility and capability to orderly assemble affinity ligands by introducing boronic acid moieties alone. By boronate conjugation via glycosylation sites, avidin was well arranged at the surface of boronic acid-decorated carbon nitride nanosheets for the assembly of biotinylated aptamers. The ordered orientation of aptamers largely relieved their inactivation caused by inter-strand entanglement, facilitating significant increase in cell affinity for the isolation of circulating tumor cells (CTCs). The reversible boronate conjugation also facilitated mild release of CTCs by acid fructose with high cell viability. This engineered interface was capable of isolating CTCs from the peripheral blood of tumor-bearing mice and cancer patients. The successful utilization of the isolated CTCs in the downstream drug susceptibility test and mutation analysis demonstrated the clinical potential of this biointerface for the early diagnosis of cancers and precision medicine.

Prognostic implication of an energy metabolism-related 11-gene signature in lung cancer.

Energy metabolism therapy has gradually shown its potential in the treatment of tumor patients, but it has significant selectivity, thus distinguishing energy subtypes of lung adenocarcinoma (LUAD) is necessary to identify patients who may benefit from energy metabolism interference therapy. Gene expression data downloaded from The Cancer Genome Atlas and Gene Expression Omnibus, molecular subtypes were selected using NMF algorithm, prognostic differentially expressed genes (DEGs) were identified with DESeq. 2 and survival package, Lasso and cox regression analysis were used to Construct of Risk Signature. The relationship between molecular subtypes and prognosis as well as clinical characteristics were evaluated. Univariate and multivariate COX regression were used to analyze the correlation between the signature and patient prognosis. Based on 592 energy metabolism-related genes, 430 LUAD samples were divided into three subtypes, of which C2 has the worst prognosis, and 942 prognostic DEGs were identified. 11-gene prognostic risk signature was constructed. Compared with the traditional clinical features of T, N, and age, this 11-gene signature performs better in predicting the risk of LUAD prognosis. At the same time, it is an independent risk factor for patient prognosis. The signature showed strong robustness in different cohorts. Compared with other published signatures, 11-gene signatures have strong clinical applicability and accuracy. The predictive signature will enable patients with LUAD to be more accurately managed in clinical practice.

Systematic profiling of diagnostic and prognostic value of autophagy-related genes for sarcoma patients.

BACKGROUND: Autophagy-related genes (ARGs) have been confirmed to have an important role in tumorigenesis and tumor microenvironment formation. Nevertheless, a systematic analysis of ARGs and their clinical significance in sarcoma patients is lacking. METHODS: Gene expression files from The Cancer Genome Atlas (TCGA) database and Genotype-Tissue Expression (GTEx) were used to select differentially expressed genes (DEGs). Differentially expressed ARGs (DEARGs) were determined by matching the DEG and HADb gene sets, which were evaluated by functional enrichment analysis. Unsupervised clustering of the identified DEARGs was conducted, and associations with tumor microenvironment (TME), immune checkpoints, and immune cells were analyzed simultaneously. Two prognostic signatures, one for overall survival (OS) and one for disease-free survival (DFS), were established and validated in an independent set. RESULTS: In total, 84 DEARGs and two clusters were identified. TME scores, five immune checkpoints, and several types of immune cells were found to be significantly different between two clusters. Two prognostic signatures incorporating DEARGs showed favorable discrimination and were successfully validated. Two nomograms combining signature and clinical variables were generated. The C-indexes were 0.818 and 0.747 for the OS and DFS nomograms, respectively. CONCLUSION: This comprehensive analyses of the ARG landscape in sarcoma showed novel ARGs related to carcinogenesis and the immune microenvironment. These findings have implications for prognosis and therapeutic responses, which reveal novel potential prognostic biomarkers, promote precision medicine, and provide potential novel targets for immunotherapy.

Utility of Magnetic Resonance Imaging in Diagnosis of Prenatal Non-Visualization of the Fetal Gallbladder: A Case-Series Study.

BACKGROUND This study aimed to assess the utility of magnetic resonance imaging (MRI) in the diagnosis of prenatal non-visualization of the fetal gallbladder (PNVGB). MATERIAL AND METHODS The clinical data of 32 pregnant women with PNVGB who underwent MRI examination during the second and third trimester of pregnancy were collected and their outcomes were analyzed. RESULTS MRI showed that 26 patients (81.3%) had isolated PNVGB and 6 (18.8%) had additional malformations. In 26 patients with isolated PNVGB, 7 were found in the gallbladder on MRI and 4 were found on subsequent ultrasonography. One patient had termination of pregnancy (TOP) and 1 patient was lost to follow-up; the remaining 24 patients were known to deliver a healthy child. Among the 6 patients with additional malformations, 3 terminated their pregnancies due to combined severe abnormalities: 1 patient with horseshoe kidney and 1 with fetal echogenic bowel both had a healthy child, while 1 with fetal growth restriction (FGR) delivered a child who walked on tiptoe. CONCLUSIONS MRI contributes to identifying PNVGB detected or suspected by ultrasonography.

Long noncoding RNA ZFAS1 promoting small nucleolar RNA-mediated 2'-O-methylation via NOP58 recruitment in colorectal cancer.

BACKGROUND: Increasing evidence supports the role of small nucleolar RNAs (snoRNAs) and long non-coding RNAs (lncRNAs) as master gene regulators at the epigenetic modification level. However, the underlying mechanism of these functional ncRNAs in colorectal cancer (CRC) has not been well investigated. METHODS: The dysregulated expression profiling of lncRNAs-snoRNAs-mRNAs and their correlations and co-expression enrichment were assessed by GeneChip microarray analysis. The candidate lncRNAs, snoRNAs, and target genes were detected by in situ hybridization (ISH), RT-PCR, qPCR and immunofluorescence (IF) assays. The biological functions of these factors were investigated using in vitro and in vivo studies that included CCK8, trans-well, cell apoptosis, IF assay, western blot method, and the xenograft mice models. rRNA 2'-O-methylation (Me) activities were determined by the RTL-P assay and a novel double-stranded primer based on the single-stranded toehold (DPBST) assay. The underlying molecular mechanisms were explored by bioinformatics and RNA stability, RNA fluorescence ISH, RNA pull-down and translation inhibition assays. RESULTS: To demonstrate the involvement of lncRNA and snoRNAs in 2'-O-Me modification during tumorigenesis, we uncovered a previously unreported mechanism linking the snoRNPs NOP58 regulated by ZFAS1 in control of SNORD12C, SNORD78 mediated rRNA 2'-O-Me activities in CRC initiation and development. Specifically, ZFAS1 exerts its oncogenic functions and significantly up-regulated accompanied by elevated NOP58, SNORD12C/78 expression in CRC cells and tissues. ZFAS1 knockdown suppressed CRC cell proliferation, migration, and increased cell apoptosis, and this inhibitory effect could be reversed by NOP58 overexpression in vitro and in vivo. Mechanistically, the NOP58 protein could be recognized by the specific motif (AAGA or CAGA) of ZFAS1. This event accelerates the assembly of SNORD12C/78 to allow for further guiding of 2'-O-Me at the corresponding Gm3878 and Gm4593 sites. Importantly, silencing SNORD12C or 78 reduced the rRNAs 2'-O-Me activities, which could be rescued by overexpression ZFAS1, and this subsequently inhibits the RNA stability and translation activity of their downstream targets (e.g., EIF4A3 and LAMC2). CONCLUSION: The novel ZFAS1-NOP58-SNORD12C/78-EIF4A3/LAMC2 signaling axis that functions in CRC tumorigenesis provides a better understanding regarding the role of lncRNA-snoRNP-mediated rRNAs 2'-O-Me activities for the prevention and treatment of CRC.

Comprehensive analysis of prognostic tumor microenvironment-related genes in osteosarcoma patients.

BACKGROUND: Tumor microenvironment (TME) plays an important role in malignant tumors. Our study aimed to investigate the effect of the TME and related genes in osteosarcoma patients. METHODS: Gene expression profiles and clinical data of osteosarcoma patients were downloaded from the TARGET dataset. ESTIMATE algorithm was used to quantify the immune score. Then, the association between immune score and prognosis was studied. Afterward, a differential analysis was performed based on the high- and low-immune scores to determine TME-related genes. Additionally, Cox analyses were performed to construct two prognostic signatures for overall survival (OS) and disease-free survival (DFS), respectively. Two datasets obtained from the GEO database were used to validate signatures. RESULTS: Eighty-five patients were included in our research. The survival analysis indicated that patients with higher immune score have a favorable OS and DFS. Moreover, 769 genes were determined as TME-related genes. The unsupervised clustering analysis revealed two clusters were significantly related to immune score and T cells CD4 memory fraction. In addition, two signatures were generated based on three and two TME-related genes, respectively. Both two signatures can significantly divide patients into low- and high-risk groups and were validated in two GEO datasets. Afterward, the risk score and metastatic status were identified as independent prognostic factors for both OS and DFS and two nomograms were generated. The C-indexes of OS nomogram and DFS nomogram were 0.791 and 0.711, respectively. CONCLUSION: TME was associated with the prognosis of osteosarcoma patients. Prognostic models based on TME-related genes can effectively predict OS and DFS of osteosarcoma patients.

Systematic Profiling of Alternative Splicing for Sarcoma Patients Reveals Novel Prognostic Biomarkers Associated with Tumor Microenvironment and Immune Cells.

BACKGROUND Alternative splicing (AS) events is a novel biomarker of tumor prognosis, but the role of AS events in sarcoma patients remains unclear. MATERIAL AND METHODS RNA-seq and clinicopathologic data of the sarcoma cohort were extracted from the TCGA database and data on AS events were downloaded from the TCGASpliceSeq database. Univariate Cox analysis, LASSO regression analysis, and multivariate Cox analysis were performed to determine the overall survival (OS)- and disease-free survival (DFS)-related AS events. Two nomograms were developed based on the independent variables, and subgroup analysis was performed. The area under the curve (AUC), calibration curve, and decision curve analysis (DCA) were used to evaluate the nomograms. Then, we used the CIBERSORT and ESTIMATE package to determine the immune cell proportion and tumor microenvironment (TME) score, respectively. The associations between AS events-based clusters and TME and immune cells were studied. RESULTS We identified 1945 and 1831 AS events as OS- and DFS-related AS events, respectively. Two nomograms based on the AS events and clinical data were established and the AUCs of nomograms ranged from 0.807 to 0.894. The calibration curve and DCA showed excellent performance of nomograms. In addition, the results indicated the distinct relationships between AS events-based clusters and OS, DFS, immune score, stromal score, and 10 immune cells. CONCLUSIONS Our study indicated that AS events are novel prognostic biomarkers for sarcoma patients that may be associated with the TME and immune cells.

Development and validation of a nomogram to predict perioperative blood transfusion in patients undergoing total knee arthroplasty.

BACKGROUND: The need for a transfusion is one of the adverse events following total knee arthroplasty (TKA), and accurately predicting this need remains challenging for arthroplasty surgeons. The purpose of the present research is to study the preoperative predictors of transfusion risk in patients following TKA and develop a nomogram. METHODS: The nomogram was developed based on a training set of 5402 patients who underwent TKA at the Affiliated Hospital of Qingdao University between September 2013 and November 2018. The independent predictors of transfusion were identified by univariate, LASSO, and binary logistic regression analyses. Then, a nomogram was established based on these independent predictors. The area under the curve (AUC), calibration curve, and decision curve analysis (DCA) were selected to evaluate the nomogram. The results were validated using an independent set of 1116 patients who underwent TKA between December 2018 and September 2019. In addition, we also carried out subgroup analyses in the training and testing sets based on the independent predictors. RESULTS: Five independent predictors were identified by multivariate analysis and were used to establish the nomogram. The AUCs of the nomogram were 0.884 (95% CI: 0.865-0.903) and 0.839 (95% CI, 0.773-0.905) in the training and testing sets, respectively. In both the training and testing sets, the calibration curve indicated that the prediction by the nomogram was highly consistent with the actual observation, and the DCA indicated that the nomogram had a favorable level of clinical usefulness. In addition, the AUC of the nomogram was significantly higher than the AUC of any independent predictor for predicting transfusion risk following TKA, and the subgroup analysis showed good performance in 20 subgroups. CONCLUSION: Lower preoperative Hb levels, simultaneous bilateral TKA, lower BMI, older age, and coronary heart disease were identified as independent predictors of postoperative transfusion in patients following TKA. A nomogram incorporating the above five predictors could accurately predict the transfusion risk.

The Timing of Symptomatic Pulmonary Embolism in Patients With Nonwarfarin Anticoagulants Following Elective Primary Total Joint Arthroplasty.

BACKGROUND: The purpose of this study is to investigate the incidence and timing of postoperative, symptomatic pulmonary embolism (PE) in patients receiving nonwarfarin treatment following primary total joint arthroplasty (TJA), to clarify the appropriate duration of postoperative VTE prophylaxis. METHODS: We retrospectively reviewed the medical records of 11,148 patients who underwent primary TJA, including total knee arthroplasty and total hip arthroplasty at our institution between January 2012 and March 2019. The median postoperative day of diagnosis of symptomatic PE and the interquartile range for day of diagnosis were determined. Multivariate Cox proportional hazards modeling was used to test the difference of timing for PE based on demographics and comorbidities. RESULTS: The overall 90-day rate of symptomatic PE was 0.71%. The median day of diagnosis for symptomatic PE was 3 days postoperatively (interquartile range, 2-7 days). Factors showed statistical significance on multivariate analysis in association with earlier timing of PE occurrence in patients with atrial fibrillation, diabetes mellitus, coronary heart disease, and history of stroke. CONCLUSION: The vast majority of symptomatic PE occurs in the early postoperative period after TJA, and atrial fibrillation, diabetes mellitus, coronary heart disease, and history of stroke were independent factors affecting the timing of symptomatic PE.

Signal transducer and activator of transcription 3 inhibition enhances vemurafenib sensitivity in colon cancers harboring the BRAFV600E mutation.

The BRAFV600E inhibitor vemurafenib is widely used to treat melanomas harboring the activated BRAFV600E mutation; however, vemurafenib showed poor efficacy in colon cancer, which impeded its clinical application for colon cancer patients with this mutation. The specific mechanism of vemurafenib resistance is not clear in colon cancer. In this study, we demonstrated that signal transducer and activator of transcription 3 (STAT3) activation influenced vemurafenib sensitivity in BRAFV600E mutant colon cancer cells. When vemurafenib was applied to two colon cancer cell lines with the BRAFV600E mutation, STAT3 was continuously activated after 6 hours. Furthermore, BCL-2 was upregulated in RKO colon cancer cells, while STAT3 remained unchanged in HT-29 colon cancer cells. This suggested that STAT3 signaling might be involved in vemurafenib sensitivity. Combining the STAT3 inhibitor STATTIC with vemurafenib further inhibited cell proliferation and promoted apoptosis by downregulating STAT3 and BCL-2 expression in RKO cells. Further studies showed that interleukin 6 (IL-6) secretion increased after RKO cells were treated with vemurafenib. STAT3 activation was induced by adding IL-6 to the supernatant, and IL-6 increased STAT3 and BCL-2 expression and antagonized vemurafenib sensitivity in HT-29 cells. Together, these results suggest that STAT3 activation maybe related to vemurafenib sensitivity in colon cancer cells, and that combining STAT3 inhibitors with vemurafenib may be a promising treatment for BRAFV600E mutant colon cancers.

Potential prognostic factors for predicting the chemotherapeutic outcomes and prognosis of patients with metastatic colorectal cancer.

OBJECTIVES: The purpose of this study was to explore whether pretreatment potential prognostic factors are related to chemotherapeutic outcomes and the prognosis of inpatients with metastatic colorectal cancer (mCRC) undergoing chemotherapy. MATERIALS AND METHODS: Data from 71 patients with mCRC were analyzed retrospectively. The relationship between the potential prognostic factors before first-line chemotherapy and the clinicopathological characteristics and chemotherapy response of the patients was calculated using Fisher's exact test and the chi-square test. The prognostic factors were analyzed using univariate and multivariate analyses. We analyzed the subgroups using the Mann-Whitney U test. RESULTS: Four factors were eventually used as prognostic factors, namely the albumin-to-globulin ratio (AGR), the fibrinogen-to-albumin ratio (FAR), the prealbumin-to-globulin ratio (PGR), and the fibrinogen-to-prealbumin ratio (FPR); the cutoff values of the four potential prognostic factors were 1.40, 10.63, 5.44, and 18.49, respectively. The high AGR and PGR groups had a higher response rate than that of the low groups. Patients in the low FAR and FPR groups showed a higher objective response rate than the high FAR and FPR groups. Patients with low FPR were associated with a higher disease control rate than patients with high FPR. Higher progression-free survival (PFS) was observed in the high AGR and PGR and low FAR and FPR groups. The AGR, FAR, PGR, and FPR were considered reliable prognostic factors for PFS in a univariate analysis. CONCLUSIONS: The prechemotherapy AGR, FAR, PGR, and FPR were good prognostic factors to predict the chemotherapy response and PFS in patients with mCRC.

Arthroscopic anatomic double-bundle ACL reconstruction using irradiated versus non-irradiated hamstring tendon allograft.

PURPOSE: To analyse the clinical outcome of anatomic double-bundle anterior cruciate ligament (ACL) reconstruction with irradiated versus non-irradiated hamstring allograft. METHODS: One hundred and twelve patients who met the inclusion and exclusion criteria of the study were prospectively randomized consecutively into irradiated hamstring tendon allograft (Ir-Allo) group and non-irradiated allograft (Non-ir-Allo) group. All surgeries were done by the same senior surgeon with double-bundle reconstruction technique. Before surgery and at follow-up points, patients were evaluated by the same observer according to clinical evaluations. RESULTS: Eighty-three patients (Non-ir-Allo, 44; Ir-Allo, 39) fulfilled complete follow-up and got full clinical evaluations. The mean follow-up period was 5.7 years (ranging from 5.0 to 6.5 years). At the final follow-up, significant differences were found when comparing the results of the two groups according to Lachman test, ADT, pivot shift test and KT-2000 arthrometer testing (P < 0.05). According to KT-2000, 86.4 % of patients in the Non-ir-Auto group and 35.9 % in the Ir-Allo group had a side-to-side difference of <3 mm. According to the overall IKDC, functional, subjective evaluations and activity level testing, no significant differences were found between the two groups. Regarding the arthritic progression, there was significant difference between the two groups (Ir-Allo group: 30.8 %, Non-ir-Allo group: 11.4 %, P < 0.05). CONCLUSION: A significant increase in anterior and rotational laxity in patients of the Ir-Allo group was found according to evaluations. No significant differences in activity level and functional scores were found between the two groups. We do not advocate the irradiated hamstring tendon allograft for ACL reconstruction. LEVEL OF EVIDENCE: I.

Combined unicompartmental knee arthroplasty and anterior cruciate ligament reconstruction in knees with osteoarthritis and deficient anterior cruciate ligament.

BACKGROUND: Relative young and more active patients with osteoarthritis (OA) of the isolated medial femorotibial compartment in conjunction with anterior cruciate ligament (ACL) deficiency are difficult to treat. The aim of this study was to explore the early clinical outcomes of combined Oxford unicompartmental knee arthroplasty (UKA) and ACL reconstruction for the patients presenting ACL deficiency and isolated OA of the medial compartment. METHODS: Twenty-eight patients were included into the study. All patients were treated by combined Oxford UKA and ACL reconstruction. Plain radiographs in the antero-posterior and lateral view and long-leg standing radiographs were routinely performed prior to and after surgery. Stress radiographs in valgus were additionally available in order to verify the well-preserved lateral compartment. The varus deformity of the knee prior to surgery and the valgus degree after surgery, the posterior slope of the tibial component and the range of motion (ROM) of the knee after surgery were measured and recorded. Clinical evaluations include Oxford Knee Score (OKS), Knee Society Score (KSS-clinical score; KSS-function score) and Tegner activity score. RESULTS: All the patients were followed up for 52 ± 8 months. The leg alignment showed 3.1 ± 0.6° of varus deformity prior to surgery and 4.0 ± 0.7° of valgus after surgery. The OKS, KSS and Tegner activity score improved significantly after surgery (P < 0.05). The mean ROM of the operated knee was 123.5 ± 2.8° at the last follow-up. The posterior slope of the tibial component was 3.9 ± 1.2°. A significant correlation was found between them according to the Pearson's correlation (r = 0.39, P = 0.03). There were 2 patients (7 %) with the complication of mobile bearing dislocation, and a second operation of replacing a thicker mobile bearing was performed for them. CONCLUSION: The early clinical data have shown that combined surgery of UKA and ACL reconstruction has revealed promising results. However, long-term follow-up studies should be done in these patients. TRIAL REGISTRATION: Current trial ISRCTN24663935 (Retrospectively registered on 21 July 2016).

Anatomic Double-Bundle Anterior Cruciate Ligament Reconstruction With a Hamstring Tendon Autograft and Fresh-Frozen Allograft: A Prospective, Randomized, and Controlled Study.

PURPOSE: To compare the clinical outcome of anatomic double-bundle (DB) anterior cruciate ligament (ACL) reconstruction with a hamstring tendon autograft versus fresh-frozen allograft. METHODS: Between January 2010 and December 2011, in a prospective randomized study, we included 157 patients who were planned to receive anatomic DB ACL reconstruction with a hamstring tendon autograft or fresh-frozen allograft. All surgeries were performed by the same senior surgeon with the DB reconstruction technique. The fixation of femoral side grafts was by means of an EndoButton, and the tibial side grafts were fixed with a bioabsorble interference screw augmented with a staple. The same rehabilitation protocol was applied to all the patients. Patients were evaluated preoperatively and at the follow-up points. Evaluations included detailed history, physical examination, radiograph, functional knee ligament testing, KT-2000 arthrometer testing, Harner's vertical jump and Daniel's one-leg hop tests, Lysholm score, Tegner score, the International Knee Documentation Committee (IKDC) standard evaluation form, and Cincinnati knee score. RESULTS: One hundred and twenty-one patients (Auto, 62; Allo, 59) fulfilled complete follow-up and got full clinical evaluations. The mean follow-up was 4.6 years (4.0 to 5.5 years) for both groups. No significant differences were found between the 2 groups according to the evaluations aforementioned except that patients in the Allo group had shorter operation time compared with the Auto group (P = .001). Fifty-three (85.5%) patients in the Auto group and 50 (84.7%) patients in the Allo group had a side-to-side difference of less than 3 mm. Four (6.5%) patients in the Auto group and 4 (6.8%) patients in the Allo group had a side-to-side difference of more than 5 mm. Fifty-nine (95.8%) patients in the Auto group and 55 (93.2%) patients in the Allo group were normal or nearly normal according to the overall IKDC. According to the subjective IKDC, the average scores were 90 and 89 points, respectively, for the Auto and Allo groups. The mean Lysholm and Tegner scores were 90 points and 7.9 points for the Auto group, respectively, and 89 points and 7.8 points for the Allo group, respectively. For the Cincinnati knee score, the average scores were 91 and 90 points, respectively, for the Auto and Allo groups. A total of 11.3% (7 of 62) of patients in the Auto group and 11.9% (7 of 59) of patients in the Allo group had an arthritic progression. There was no statistical difference between the 2 groups at the final follow-up. CONCLUSIONS: With the anatomic DB ACL reconstruction technique, comparable objective and subjective clinical results can be achieved with the use of a fresh-frozen hamstring tendon allograft compared with an autograft. LEVEL OF EVIDENCE: Level II, prospective randomized clinical trial.

Mechanical and IL-1ß Responsive miR-365 Contributes to Osteoarthritis Development by Targeting Histone Deacetylase 4.

Mechanical stress plays an important role in the initiation and progression of osteoarthritis. Studies show that excessive mechanical stress can directly damage the cartilage extracellular matrix and shift the balance in chondrocytes to favor catabolic activity over anabolism. However, the underlying mechanism remains unknown. MicroRNAs (miRNAs) are emerging as important regulators in osteoarthritis pathogenesis. We have found that mechanical loading up-regulated microRNA miR-365 in growth plate chondrocytes, which promotes chondrocyte differentiation. Here, we explored the role of the mechanical responsive microRNA miR-365 in pathogenesis of osteoarthritis (OA). We found that miR-365 was up-regulated by cyclic loading and IL-1ß stimulation in articular chondrocytes through a mechanism that involved the transcription factor NF-κB. miR-365 expressed significant higher level in rat anterior cruciate ligament (ACL) surgery induced OA cartilage as well as human OA cartilage from primary OA patients and traumatic OA Patients. Overexpression of miR-365 in chondrocytes increases gene expression of matrix degrading enzyme matrix metallopeptidase 13 (MMP13) and collagen type X (Col X). The increase in miR-365 expression in OA cartilage and in response to IL-1 may contribute to the abnormal gene expression pattern characteristic of OA. Inhibition of miR-365 down-regulated IL-1ß induced MMP13 and Col X gene expression. We further showed histone deacetylase 4 (HDAC4) is a direct target of miR-365, which mediates mechanical stress and inflammation in OA pathogenesis. Thus, miR-365 is a critical regulator of mechanical stress and pro-inflammatory responses, which contributes cartilage catabolism. Manipulation of the expression of miR-365 in articular chondrocytes by miR-365 inhibitor may be a potent therapeutic target for the prevention and treatment of osteoarthritis.

SIRT4 Has an Anti-Cancer Role in Cervical Cancer by Inhibiting Glutamine Metabolism via the MEK/ERK/C-Myc Axis.

BACKGROUND/AIM: Glutamine metabolism is crucial in cell proliferation, aging, and apoptosis across various cancer types. Existing research indicates that Sirtuin 4 (SIRT4), primarily located in mitochondria, modulates this process. This study aimed to clarify the regulatory relationship between SIRT4 and glutamine metabolism in cervical cancer. MATERIALS AND METHODS: SIRT4 mRNA levels and their clinical correlation to cervical cancer were analyzed using the UALCAN database. Immunohistochemistry (IHC) was performed to assess SIRT4 protein expression in tissue samples from cervical cancer patients. Transient transfection was employed to create Hela and Siha cell lines with overexpressed SIRT4, mitogen-activated extracellular signal-regulated kinase (MEK), and glutaminase 1 (GLS1). The impact on cellular functions was studied using MTT, soft agar, transwell, and western blotting assays. Glutamate and ATP levels were also measured to evaluate metabolic changes. RESULTS: Low levels of SIRT4 mRNA in cervical cancer tissues correlated with tumor metastasis and poor survival rates. Overexpression of SIRT4 led to suppressed cell proliferation, colony growth, and motility, along with significant down-regulation of GLS expression, a key contributor to glutamine metabolism. Additionally, SIRT4 overexpression resulted in the inactivation of the MEK/ERK/c-myc signaling pathway, while overexpression of MEK reversed these effects. Notably, the inhibitory effects of SIRT4 on cell proliferation, colony formation, migration, and invasion in Hela and Siha cells were significantly attenuated following GLS1 overexpression. CONCLUSION: SIRT4 acts as an anti-cancer agent in cervical cancer by inhibiting glutamine metabolism through the MEK/ERK/c-myc signaling pathway, providing a novel sight for cervical cancer therapy.

CEBPB-mediated upregulation of SERPINA1 promotes colorectal cancer progression by enhancing STAT3 signaling.

Colorectal cancer (CRC) is a highly malignant carcinoma associated with poor prognosis, and metastasis is one of the most common causes of death in CRC. Serpin Family A Member 1 (SERPINA1) is a serine protease inhibitor from the Serpin family. Till now, the function and mechanism of SERPINA1 in CRC progression have not been fully illustrated. We established highly metastatic colorectal cancer cells named as RKO-H and Caco2-H by mice liver metastasis model. By integrative bioinformatic approaches, we analyzed the prognostic value and clinical significance of SERPINA1 in CRC, and predicted potential transcription factors. Colony formation, EDU, MTS, Transwell and wound healing assay were performed to evaluate the biological functions of SERPINA1 in CRC in vitro. Experiments in vivo were conducted to explore the effects of SERPINA1 on liver metastasis of CRC. ChIP and luciferase reporter gene assays were performed to identify the transcriptional regulatory mechanism of SERPINA1 by CEBPB. Our results show that SERPINA1 is highly expressed in CRC and correlated with poor clinical outcomes. SERPINA1 promotes the proliferation, migration by activating STAT3 pathway. Mechanistically, CEBPB binds SERPINA1 gene promoter sequence and promotes the transcription of SERPINA1. SERPINA1 drives CEBPB-induced tumor cell growth and migration via augmenting STAT3 signaling. Our results suggest that SERPINA1 is a potential prognostic marker and may serve as a novel treatment target for CRC.

The value of sequential application of hydrogen peroxide, povidone-iodine and physiological saline in reducing postoperative infections after total knee arthroplasty: A prospective, randomized, controlled study.

BACKGROUND: Currently, in the field of total joint arthroplasty (TJA), there are no studies that have demonstrated the value of the sequential application of hydrogen peroxide, povidone-iodine, and physiological saline during the surgical procedure in decreasing postoperative infections in total knee arthroplasty (TKA), and in decreasing the incidence of periprosthetic joint infections (PJI) in particular. This study aimed to assess the efficacy of the sequential application of hydrogen peroxide, povidone-iodine, and physiological saline in reducing postoperative infections in TKA. METHODS: The study prospectively included 4743 patients, with Group A (2371, 49.9%) receiving sequential intraoperative application of hydrogen peroxide, povidone-iodine, and physiological saline irrigation of the incision, and Group B (2372, 50.1%) receiving intraoperative application of physiological saline irrigation of the incision only, to collect the patients' baseline data and clinical characteristics, and to statistically assess the incidence of superficial infections and the PJI during the follow up period to evaluate the clinical value of the study. RESULTS: The baseline levels of patients in Groups A and B were comparable. There were 132 (2.8%) lost visits during the study period. The incidence of superficial infections within 30 days after surgery was 0.22% in Group A and 1.17% in Group B, the difference between the two groups was statistically significant (P = 0.007). The incidence of PJI was 0.17% in Group A and 1.26% in Group B, the difference between the two groups was statistically significant (P = 0.0121). CONCLUSION: Sequential application of hydrogen peroxide, povidone-iodine, and physiological saline to irrigate incision in TKA can significantly reduce the incidence of postoperative superficial infections and PJI. The scientific and rational application of this therapy intraoperatively greatly reduces the incidence of PJI and postoperative superficial infections, which is of great benefit to the patient's prognosis.