Identification and Verification of Novel Biomarkers Involving Rheumatoid Arthritis with Multimachine Learning Algorithms: An In Silicon and In Vivo Study.

Background: Rheumatoid arthritis (RA) remains one of the most prevalent chronic joint diseases. However, due to the heterogeneity among RA patients, there are still no robust diagnostic and therapeutic biomarkers for the diagnosis and treatment of RA. Methods: We retrieved RA-related and pan-cancer information datasets from the Gene Expression Omnibus and The Cancer Genome Atlas databases, respectively. Six gene expression profiles and corresponding clinical information of GSE12021, GSE29746, GSE55235, GSE55457, GSE77298, and GSE89408 were adopted to perform differential expression gene analysis, enrichment, and immune component difference analyses of RA. Four machine learning algorithms, including LASSO, RF, XGBoost, and SVM, were used to identify RA-related biomarkers. Unsupervised cluster analysis was also used to decipher the heterogeneity of RA. A four-signature-based nomogram was constructed and verified to specifically diagnose RA and osteoarthritis (OA) from normal tissues. Consequently, RA-HFLS cell was utilized to investigate the biological role of CRTAM in RA. In addition, comparisons of diagnostic efficacy and biological roles among CRTAM and other classic biomarkers of RA were also performed. Results: Immune and stromal components were highly enriched in RA. Chemokine- and Th cell-related signatures were significantly activated in RA tissues. Four promising and novel biomarkers, including CRTAM, PTTG1IP, ITGB2, and MMP13, were identified and verified, which could be treated as novel treatment and diagnostic targets for RA. Nomograms based on the four signatures might aid in distinguishing and diagnosing RA, which reached a satisfactory performance in both training (AUC = 0.894) and testing (AUC = 0.843) cohorts. Two distinct subtypes of RA patients were identified, which further verified that these four signatures might be involved in the immune infiltration process. Furthermore, knockdown of CRTAM could significantly suppress the proliferation and invasion ability of RA cell line and thus could be treated as a novel therapeutic target. CRTAM owned a great diagnostic performance for RA than previous biomarkers including MMP3, S100A8, S100A9, IL6, COMP, LAG3, and ENTPD1. Mechanically, CRTAM could also be involved in the progression through immune dysfunction, fatty acid metabolism, and genomic instability across several cancer subtypes. Conclusion: CRTAM, PTTG1IP, ITGB2, and MMP13 were highly expressed in RA tissues and might function as pivotal diagnostic and treatment targets by deteriorating the immune dysfunction state. In addition, CRTAM might fuel cancer progression through immune signals, especially among RA patients.

A Sternoclavicular Joint-Specific Plate for the Displaced Medial-End Clavicle Fracture.

Objectives: This study aimed to introduce a sternoclavicular joint (SCJ)-specific plate for the treatment of medial-end clavicle fracture and evaluate the clinical and radiological results of this method. Methods: From January 2006 to December 2020, 31 patients with displaced medial-end clavicle fractures were included in this study, with 8 patients with accompanying SCJ dislocation. Abduction and forward elevation of the shoulder, the Visual Analogue Scale (VAS), and the American Shoulder and Elbow Surgeons Score (ASES) were used for evaluation before index surgery and at the latest follow-up. Results: After an average of 98.5 (range, 13 to 171) months, the mean VAS significantly decreased from 6.8 ± 1.0 preoperatively to 0.9 ± 0.8 at the latest follow-up (P < 0.001). The mean ASES score significantly increased from 34.3 ± 7.8 preoperatively to 90.2 ± 4.9 at the latest follow-up (P < 0.001). The mean abduction of the shoulder significantly increased from 72.1 ± 6.6 preoperatively to 169.5 ± 8.5 at the latest follow-up (P < 0.001). The mean forward elevation of the shoulder significantly increased from 97.1 ± 11.0 preoperatively to 163.1 ± 11.5 at the latest follow-up (P < 0.001). The union of all fractures was achieved, and all implants were removed. No loose or breakage of implants was observed. No vascular or nerve damage occurred during the operation. Conclusions: This SCJ-specific plate provided excellent long-term results for the treatment of displaced medial-end clavicle fractures and was an alternative implant for medial-end clavicle fractures with or without small or comminuted medial fragments, especially those associated with SCJ dislocation.

A Novel Mutation of the KLK6 Gene in a Family With Knee Osteoarthritis.

To investigate the correlation between gene mutation and knee osteoarthritis (KOA), a whole-exome sequencing (WES) was applied to analyze blood samples of four KOA patients and two normal subjects in a family. Gene mutations were identified by gene-trapping and high-throughput sequencing analysis across the differences between the patients and normal subjects. The interactive gene network analysis on the retrieval of interacting genes (STRING) database and the KOA-related genes expression data sets was performed. A possibly detrimental and nonsynonymous mutation at the kallikrein-related peptidase 6 (KLK6) gene (rs201586262, c. C80A, P27H) was identified and attracted our attention. KLK6 belongs to the kallikrein family of serine proteases and its serum level is known as a prevalent biomarker in inflammatory and malignant diseases. KLK6 expresses in the extracellular compartment for matrix degradation, highlighting that KLK6 plays a role in the pathogenesis of KOA. By using the gene databases, the KOA-related genes were mined after de-duplication and IL6 was selected as the most relevant gene through interactive analysis of protein-protein interaction (PPI) network. The data suggested that KLK6 gene mutation and the related expression alteration of IL6 gene might determine the occurrence of hereditary KOA. The is the first study discovering the gene mutation of KLK6 as a factor of pathogenesis of KOA, especially the hereditary KOA.

Long-term results for traumatic sternoclavicular joint dislocation treated with a sternoclavicular joint-specific plate.

BACKGROUND: The treatment of traumatic sternoclavicular joint (SCJ) dislocation (SCJD) with internal fixation has been reported with good short-term results, but data on its long-term results are scarce. METHODS: Patients with traumatic SCJD treated with an SCJ-specific plate between January 2003 and January 2018 were evaluated retrospectively. All implants were routinely removed. Data from radiography, the American Shoulder and Elbow Surgeons score, the visual analogue scale and abduction and forward elevation of the shoulder were collected and evaluated before the index surgery, at implant removal and at the latest follow-up. RESULTS: A total of 22 patients were included in this study with a mean follow-up period of 94.8 months. All patients maintained good reduction after the index surgery and implant removal. The visual analogue scale significantly improved from 7.1 ± 1.3 before the index surgery to 0.9 ± 1.0 at implant removal (P < 0.001) and to 1.0 ± 1.1 at the latest follow-up (P < 0.001); the American Shoulder and Elbow Surgeons score significantly improved from 37.9 ± 10.1 to 90.8 ± 7.8 (P < 0.001) and to 86.7 ± 8.6; and both abduction and forward elevation of the shoulder significantly improved at the latest follow-up (P < 0.001). There was no significant difference in the clinical results after implant removal. CONCLUSION: Traumatic SCJD treated with an SCJ-specific plate appeared to be efficient, with satisfactory clinical and radiological results at long-term follow-up.

Growth factors-based beneficial effects of platelet lysate on umbilical cord-derived stem cells and their synergistic use in osteoarthritis treatment.

Poor viability of mesenchymal stem cells (MSCs) at the transplanted site often hinders the efficacy of MSCs-based therapy. Platelet lysate (PL) contains rich amounts of growth factors, which benefits cell growth. This study aimed to explore how human PL benefits umbilical cord-derived MSCs (huc-MSCs), and whether they have synergistic potential in osteoarthritis (OA) treatment. As quality control, flow cytometry and specific staining were performed to identify huc-MSCs, and ELISA was used to quantify growth factors in PL. CCK-8 and flow cytometry assays were performed to evaluate the effects of PL on the cell viability and cell cycle progression of huc-MSCs. Wound healing and transwell assays were conducted to assess the migration of huc-MSCs. RNA sequencing, real time PCR, and Western blot assays were conducted to explore the growth factors-based mechanism of PL. The in vitro results showed that PL significantly promoted the proliferation, cell cycle, and migration of huc-MSCs by upregulating relevant genes/proteins and activating beclin1-dependent autophagy via the AMPK/mTOR signaling pathway. The main growth factors (PDGF-AA, IGF-1, TGF-ß, EGF, and FGF) contributed to the effects of PL in varying degrees. The in vivo data showed that combined PL and huc-MSCs exerted significant synergistic effect against OA. The overall study determined the beneficial effects and mechanism of PL on huc-MSCs and indicated PL as an adjuvant for huc-MSCs in treating OA. This is the first report on the growth factors-based mechanism of PL on huc-MSCs and their synergistic application. It provides novel knowledge of PL's roles and offers a promising strategy for stem cell-based OA therapy by combining PL and huc-MSCs.

Lateral bone flap approach for displaced intra-articular calcaneus fractures.

BACKGROUND: The traditional extended lateral approach for calcaneus fractures can provide sufficient exposure for reduction, but complications are frequent. The minimally invasive approach does limited damage to the soft tissue and its complication rate is low, but provides limited surgical exposure for complicated fracture. Thus, an approach that could provide wide exposure with less soft tissue injury is important. METHODS: The lateral bone flap approach, we proposed, involved the same incision as the extended lateral approach. After incision, osteotomy was performed around the lateral bulged wall of the calcaneus without dissection of soft tissue. The lateral wall was free from calcaneus and connected with the soft tissue, and the lateral bone flap was developed. The reduction of fracture and the fixation of plate were performed as usual. Next, the bone flap was reduced and sutured. This retrospective study of lateral bone flap approach included 63 cases of 58 patients with displaced intra-articular calcaneus fractures from January 2011 to January 2015. Clinical and radiological outcomes and complications were all recorded. RESULTS: Radiological outcome was significantly improved at 3 months and 2 years post-operatively compared with that of pre-operatively (P < 0.01). The Maryland Foot Score at the last follow up was 87.2 ± 7.0. The excellent/good rate was 90.5%. One case of delayed wound healing occurred. No infection or sural nerve injury occurred. CONCLUSION: The lateral bone flap approach is simple, safe and effective for displaced intra-articular calcaneus.

Cell-free plasma hypermethylated CASZ1, CDH13 and ING2 are promising biomarkers of esophageal cancer.

Identifying sensitive and specific biomarkers for early detection of cancer is immensely imperative for early diagnosis and treatment and better clinical outcome of cancer patients. This study aimed to construct a specific DNA methylation pattern of cancer suppressor genes and explore the feasibility of applying cell-free DNA based methylation as a biomarker for early diagnosis of esophageal squamous cell carcinoma (ESCC). We recruited early stage ESCC patients from Yangzhong County, China. The Illumina Infinium 450K Methylation BeadChip was used to construct a genome-wide DNA methylation profile. Then, differentiated genes were selected for the validation study using the Sequenom MassARRAY platform. The frequency of methylation was compared between cancer tissues, matched cell-free DNAs and normal controls. The specific methylation profiles were constructed, and the sensitivity and specificity were calculated. Seven CG sites in three genes CASZ1, CDH13 and ING2 were significantly hypermethylated in ESCC as compared with normal controls. A significant correlation was found between the methylation of DNA extracted from cancer tissues and matched plasma cell-free DNA, either for individual CG site or for cumulative methylation analysis. The sensitivity and specificity reached 100% at an appropriate cut-point using these specific methylation biomarkers. This study revealed that aberrant DNA methylation is a promising biomarker for molecular diagnosis of esophageal cancer. Hypermethylation of CASZ1, CDH13 and ING2 detected in plasma cell-free DNA can be applied as a potential noninvasive biomarker for diagnosis of esophageal cancer.

Theabrownin triggers DNA damage to suppress human osteosarcoma U2OS cells by activating p53 signalling pathway.

Osteosarcoma becomes the second leading cause of cancer death in the younger population. Current outcomes of chemotherapy on osteosarcoma were unsatisfactory to date, demanding development of effective therapies. Tea is a commonly used beverage beneficial to human health. As a major component of tea, theabrownin has been reported to possess anti-cancer activity. To evaluate its anti-osteosarcoma effect, we established a xenograft model of zebrafish and employed U2OS cells for in vivo and in vitro assays. The animal data showed that TB significantly inhibited the tumour growth with stronger effect than that of chemotherapy. The cellular data confirmed that TB-triggered DNA damage and induced apoptosis of U2OS cells by regulation of Mki67, PARP, caspase 3 and H2AX, and Western blot assay showed an activation of p53 signalling pathway. When P53 was knocked down by siRNA, the subsequent downstream signalling was blocked, indicating a p53-dependent mechanism of TB on U2OS cells (p53 wt). Using osteosarcoma cell lines with p53 mutations (HOS, SAOS-2 and MG63), we found that TB exerted stronger inhibitory effect on U2OS cells than that on p53-mut cell lines, but it also exerted obvious effect on SAOS-2 cells (p53 null), suggesting an activation of p53-independent pathway in the p53-null cells. Interestingly, theabrownin was found to have no toxicity on normal tissue in vivo and could even increase the viability of p53-wt normal cells. In sum, theabrownin could trigger DNA damage and induce apoptosis on U2OS cells via a p53-dependent mechanism, being a promising candidate for osteosarcoma therapy.

Analysis of differentially expressed genes in white blood cells isolated from patients with major burn injuries.

The aim of the present study was to identify differentially expressed genes (DEGs) and their related functions and pathways of major burn injuries, and to prevent the occurrence of complications. The expression profiling of E-GEOD-37069 was downloaded from ArrayExpress Archive. The DEGs of major burn injuries were identified. Gene Ontology (GO) and Kyoto Encyclopedia of Gene and Genomes (KEGG) functional enrichment analysis were then performed for the DEGs. Based on the KEGG database, a pathway relationship network was constructed, and DEGs in significant GO terms and pathways were investigated. Gene signal network and gene co-expression network of these inserted DEGs were constructed. A total of 3,328 DEGs of major burn injuries were identified, including 1,337 up- and 1,991 downregulated DEGs. These DEGs were mainly enriched into various GO terms, including transcription, DNA-dependent, signal transduction and blood coagulation. Moreover, they were also enriched into different pathways, such as hematopoietic cell lineage, metabolic pathway and chemokine signaling pathway. The pathway relationship network was constructed with 72 nodes. The MAPK signaling pathway was the hub node. Based on the same gene symbol, 702 DEGs were obtained, identified in both GO terms and pathways. Finally, the gene signaling network and gene co-expression network were constructed with 391 and 128 nodes, respectively. These identified DEGs, including GNB2, LILRA2, ARRB2 and ARHGEF2, may be potential key genes involved in the treatment of major burn injuries and prevention of complications.

Mid-term results of Oxford phase-3 medial unicompartmental knee arthroplasty for medial arthritis in Chinese patients.

BACKGROUND: The purpose of this study was to evaluate the mid-term results of an Oxford phase-3 unicompartmental knee arthroplasty (UKA) for medial arthritis in Chinese patients. METHODS: The study included 64 patients who underwent a minimally invasive Oxford phase-3 UKA for medial knee arthritis. The patients were clinically evaluated preoperatively and at the final follow-up according to the clinical and functional components of the Knee Society Score (KSS), the Hospital for Special Surgery knee score and range of motion. A Kaplan-Meier survivorship analysis was performed with revision surgery as the end point. RESULTS: The mean preoperative clinical KSSs increased from 63.2 to 91.4 post-operatively, and the mean functional KSSs increased from 54.9 to 86.5 post-operatively. In addition, the mean Hospital for Special Surgery scores increased from 59.5 to 86.4. The mean active knee flexion increased from 109.1° preoperatively to 123.6° post-operatively. A total of six patients (six knees) required revision surgery at the time of the maximum 10-year follow-up. Four conversions to total knee arthroplasty were performed because of arthritis progression in the lateral compartment. One revision to total knee arthroplasty was performed for aseptic loosening, and one liner exchange was performed for wear. The cumulative survival rates at the 6- and 8-year follow-ups were 97% and 93%, respectively. CONCLUSION: Oxford phase-3 UKA was largely applicable for medial arthritis in Chinese patients. However, the Oxford phase-3 medial UKA selection criteria for young Chinese males need further exploration to obtain the best treatment effect.

Dumbbell-Shaped Epidural Cavernous Hemangioma in the Thoracic Spine Mimicking Schwannoma.

Dumbbell-shaped epidural cavernous hemangiomas (CHs) are extremely rare, and they are easily misdiagnosed as spinal schwannomas. Herein, the authors report 1 rare case of dumbbell-shaped epidural CH in the thoracic spine. To the best of our knowledge, only a few cases of dumbbell-shaped epidural CHs in thoracic spine have been reported. Furthermore, the clinical characteristics and treatments for spinal epidural CHs were investigated and reviewed.

Lentivirus-mediated RNA interference targeting UbcH10 reduces cell growth and invasion of human osteosarcoma cells via inhibition of Ki-67 and matrix metalloproteinases.

Osteosarcoma (OS) is the most commonly diagnosed primary malignancy affecting the bone. UbcH10 is a cancer-related E2-ubiquitin-conjugating enzyme. An overexpression of UbcH10 is significantly associated with tumor grade and cellular proliferation. However, limited evidence exists with regard to the biological function of UbcH10 in OS. The present study created a UbcH10 knockdown OS cell line using lentivirus-mediated RNA interference. The expression of UbcH10 was significantly reduced in UbcH10-targeted small hairpin RNA-expressing lentivirus OS cells. The downregulation of UbcH10 suppressed OS cell proliferation and colony formation ability via decreased Ki-67 expression. UbcH10 knockdown OS cells exhibited impaired invasion and migration abilities. Furthermore, knockdown of UbcH10 led to decreased levels of matrix metalloproteinase-3 and -9 in OS cells. The present study demonstrated the role of UbcH10 in OS cell proliferation, invasion and migration, which suggests that UbcH10 may be a potential candidate for OS therapy.

Administration of enoxaparin 24 h after total knee arthroplasty: safer for bleeding and equally effective for deep venous thrombosis prevention.

BACKGROUND: The purpose of this study was to determine the safety and effectiveness, as assessed by risk of bleeding and incidence of deep venous thrombosis (DVT), of administering delayed low-molecular-weight heparin (LMWH) after total knee arthroplasty. METHODS: A prospective study of 210 consecutive patients undergoing primary unilateral total knee arthroplasty was undertaken. The patients were randomized into two groups: one of which was managed according to a standard LMWH program (LMWH-s group) and the other with delayed LMWH (LMWH-p). LMWH was initiated 12 h after wound closure in the LMWH-s group, and 24 h after wound closure in the LMWH-p group. RESULTS: The total blood loss in the first three postoperative days was calculated and all complications were recorded. The mean total blood loss was 435 and 387 mL in the LMWH-s group and LMWH-p group, respectively (p < 0.01). No significant difference in the incidence of symptomatic DVT was observed. The mean length of hospital stay was 7.29 days in the LMWH-s group and 6.56 days in the LMWH-p group (p < 0.05). CONCLUSIONS: After total knee arthroplasty, LMWH-p is safer for bleeding than LMWH-s and equally effective concerning prevention of DVT.

Patella rings for treatment of patellar fracture.

To evaluate a new fixation technique for patellar fracture using patella rings. A total of 75 patients (average age of 51.3 years) with comminuted or transverse patellar fractures were treated by fixation with patella rings. The Böstman scores at the time of bone union and at 12 months postoperatively were recorded, as was the degree of pain on a visual analogue scale (VAS), the range of motion of the knee at 12 months postoperatively, and any signs of postoperative complications. The average Böstman scores for patients in the transverse fracture group were 25.2 and 29.4 at 3 and 12 months postoperatively, respectively, while the scores for patients in the comminuted fracture group were 27.6 and 28.7, at the same time points. Böstman scores were graded as excellent and good in more than 90 % for patients with either a transverse or comminuted fracture. At the time of 12 months after surgery, the VAS score for patients with comminuted fractures was 0.38, whereas the score for patients in the transverse fracture group was 0.35. No statistically significant difference was found in the range of motion between the affected and uninjured knee at 12 months after surgery for patients in both groups (P > 0.05). This new fixation technique using a patella ring resulted in good outcomes for both transverse fracture and comminuted fracture and is beneficial for patients wishing to commence early functional activity.

Intra-discal vancomycin-loaded PLGA microsphere injection for MRSA discitis: an experimental study.

OBJECTIVE: To prepare the vancomycin hydrochloride (VA)-loaded poly lactic acid-glycolic acid (PLGA) copolymer microsphere by the multiple emulsion method and evaluate its therapeutic effects on infective discitis. METHODS: Firstly, the particle diameter distribution, shape, encapsulation efficiency, drug-loaded dosage and release curve of VA-PLGA microspheres were evaluated in vitro. Rabbits with methicillin-resistant Staphylococcus aureus infective discitis were treated with VA-PLGA intra-discal injection. Meanwhile, VA intravenous injection, blank PLGA microspheres intra-discal injection served as controls. Thirty days later, therapeutic effects were evaluated through X-ray radiophotography, histopathological and bacteriological examination. RESULTS: Mean particle diameter was between 61.57 ± 4.37 and 67.45 ± 8.13 µm, and mean encapsulation efficiency was between 60.20 ± 1.61 and 75.27 ± 1.60 %m/m. In vitro release experiment showed that the release time was over 30 days. The result of in vivo experiment showed that inflammatory reaction in the VA-PLGA intra-discal injection group was milder than the intravenous injection group (P < 0.05), also with less inflammation. The bacterial count was also significantly lower (1.02 × 10(3) ± 1.22 × 10(3) CFU/g) than the intravenous injection group (7.51 × 10(4) ± 7.16 × 10(4) CFU/g) (P < 0.05). Besides these data, the amount used in VA-PLGA intra-discal injection group is about 20 mg, and that used in the intravenous injection group is about 2.4 g. So, we just use 1/120 of VA i.v. to obtain the better results with our microparticles. CONCLUSION: Intra-discal injection with VA-PLGA sustained-release microspheres can use much less dosage, and effectively control and reduce infective discitis, and the therapeutic effect is superior to that of intravenous injection. A need for the clinical trials will be carried out in the near future.

Co and Cr accumulation in hair after metal-on-metal hip resurfacing arthroplasty.

BACKGROUND: Currently, it is unclear whether Co and Cr levels are persistently elevated in the body after hip resurfacing arthroplasty (HRA). This study aimed to evaluate Co and Cr levels in the body after HRA using hair as samples. METHODS: Among 22 patients who had metal-on-metal HRA(m-o-m group) using the DePuy ASR system, scalp hair of 1.5 cm in length and 2 g in weight was collected before and at 6 and 12 months after surgery. Inductively coupled plasma-mass spectrometry (ICP-MS) was used to detect Co and Cr levels in the scalp hair. Another 22 patients with total hip replacements were randomly selected from a similar age range as controls (m-o-p group). Metal levels in hair were detected with multiple measurements and analysed by repeated measures analysis of variance. RESULTS: Both Co and Cr levels were significantly increased in the m-o-m group after surgery compared with those in the m-o-p group (P < 0.01). Co levels at 6 months post-operatively were 12-fold those before surgery, and Co levels 12 months postoperatively were lower than those at 6 months, but this difference was not significant (P = 0.0805) between the two time points in the m-o-m group. Cr levels were persistently elevated after surgery and were increased by 10-fold compared with those at baseline in the m-o-m group. CONCLUSIONS: Co and Cr levels in scalp hair are ideal indicators for metal levels in the body after HRA. Co and Cr levels in the body were greatly increased after metal-on-metal HRA. Long-term accumulation of these metals needs to be further investigated