G3BP1 coordinates lysophagy activity to protect against compression-induced cell ferroptosis during intervertebral disc degeneration.

Lysophagy is a form of selective autophagy to remove unwanted lysosomes. However, its role in the pathogenesis of intervertebral disc degeneration (IDD) remains unclear. We intended to investigate the relationship between lysophagy and ferroptosis, as well as the potential involved molecules during IDD. Human nucleus pulposus (NP) cells were obtained from clinical patients. The protein levels, protein colocalization and cellular reactive oxygen species levels were assessed by western blotting, immunofluorescence analysis, immunoprecipitation and flow cytometry, respectively. The in vivo experiments were conducted based on the needle puncture-induced IDD model in rats. Compression pressure induces the lysophagy inactivation and lysosomal damage, resulting in iron overload and ferroptosis in human NP cells. Notably, Ras GTPase-activating protein-binding proteins 1 (G3BP1) resides at lysosomes to coordinate lysophagy activity mainly via the function of G3BP1/TSC2 complex. Dysfunction of G3BP1/TSC2 complex accelerates the lysosomal damage and ferroptosis in NP cells. Besides, inhibition of mTOR signalling ameliorates lysosomal damage and protects against cell ferroptosis. The in vivo experiments also demonstrate that the G3BP1/mTOR signalling is involved in the progression of IDD. These findings illustrate the relationship between lysophagy and compression-induced cell ferroptosis. It also indicates the positive role of G3BP1 and may provide potential targets for IDD treatment.

Modified arthroscopic Brostr?m procedure for the treatment of chronic ankle instability combined with multiple ligament laxity / 中华创伤杂志

Objective:To investigate the effectiveness of modified arthroscopic Brostr?m procedure for the treatment of chronic ankle instability combined with multiple ligament laxity.Methods:A retrospective case series study was used to analyze the clinical data of 26 patients with chronic ankle instability combined with multiple ligament laxity treated at Union Hospital, Tongji Medical College of Huazhong University of Science and Technology from January 2016 to December 2020, including 10 males and 16 females; aged 18-48 years [(27.5±7.1)years]. All patients underwent arthroscopic repair of the anterior talofibular ligament (ATFL) by the modified Brostr?m procedure. Healing of surgical incisions was observed after operation. The change of talus tilt angle for ankle stability evaluation, the American Orthopedic Foot and Ankle Society (AOFAS) ankle-hindfoot score for ankle function evaluation, and the visual analogue score (VAS) for pain evaluation were assessed before operation, at 3 months postoperatively and at the last follow-up. Complications were observed.Results:All patients were followed up for 18-47 months [(25.3±8.5)months]. All surgical incisions were healed at stage I. The talus tilt angle was decreased from preoperative (15.6±4.7)° to (4.1±1.3)° and (3.5±0.9)° at 3 months postoperatively and at the last follow-up (all P<0.01). The AOFAS ankle-hindfoot score was improved from preoperative (65.8±14.5)points to (86.5±5.6)points and (93.4±4.2)points at 3 months postoperatively and at the final follow-up (all P<0.01). The VAS was decreased from preoperative 3.0 (2.0, 4.0)points to 1.5 (0.0, 2.0)points and 1.0 (0.0, 1.2)points at 3 months postoperatively and at the last follow-up (all P<0.01). Significantly higher AOFAS ankle-hindfoot score and lower VAS were found at the final follow-up when compared with the scores at 3 months postoperatively (all P<0.05). One patient developed superficial peroneal nerve injury, which was recovered spontaneously without special treatment. Conclusion:For chronic ankle instability combined with multiple ligament laxity, the modified arthroscopic Brostr?m procedure has advantages of improved ankle stability, good ankle function recovery, obvious pain relief and less postoperative complications.

Arthroscopic medullary decompression combined with platelet-rich plasma therapy for bone marrow edema of the talus / 中华创伤杂志

Objective:To report the efficacy of arthroscopic medullary decompression combined with platelet-rich plasma (PRP) in the treatment of bone marrow edema of the talus.Methods:A retrospective case series study was used to analyze the clinical data of 17 patients with bone marrow edema of the talus admitted to Union Hospital, Tongji Medical College, Huazhong University of Science and Technology from January 2018 to July 2020. There were 11 males and 6 females, with the age range of 15-56 years [(45.7±4.3)years]. All patients were subjected to arthroscopic medullary decompression combined with the administration of PRP. Operation time and wound healing were recorded. Maximum area of bone marrow edema was measured by MRI preoperatively and at 6 and 12 months postoperatively. Ankle range of motion (ROM), visual analog score (VAS) and American Association of Foot and Ankle Surgery (AOFAS) ankle-hindfoot score were measured preoperatively and at 6 and 12 months postoperatively. Complications were also detected.Results:All patients were followed up for 12-41 months [(16.7±2.1)months]. Operation time was 45.2-68.5 minutes [(53.4±12.4)minutes]. All wounds were healed at stage I. The maximum area of bone marrow edema decreased from (28.2±6.9)mm 2 preoperatively to (16.3±5.7)mm 2 at 6 months postoperatively and to (7.1±1.7)mm 2 at 12 months postoperatively (all P<0.01). Ankle ROM increased from (52.2±8.9)° preoperatively to (72.3±3.1)° at 6 months postoperatively and to (83.1±2.8)° at 12 months postoperatively (all P<0.01). VAS decreased from (8.2±0.6)points preoperatively to (6.5±0.4)points at 6 months postoperatively and to (3.1±0.8)points at 12 months postoperatively (all P<0.01). AOFAS ankle-hindfoot score increased from (32.4±4.8)points preoperatively to (54.4±6.5)points at 6 months postoperatively and to (88.7±4.3)points at 12 months postoperatively (all P<0.01). There were significant differences in maximum area of bone marrow edema of the talus, ankle ROM, VAS and AOFAS ankle-hindfoot score at 12 months postoperatively when compared with those at 6 months postoperatively (all P<0.01). One patient showed the symptom of localized skin numbness postoperatively, and improved with nerve nutrition therapy. Conclusion:Arthroscopic medullary decompression combined with PRP therapy for bone marrow edema of the talus presents good short-term clinical benefits in terms of reduced extent of bone marrow edema, improved ankle ROM, attenuated pain, improved ankle joint function and few postoperative complications.

Comparison of arthroscopy-assisted reduction and internal fixation versus open reduction and internal fixation for tibial plateau fractures / 中华创伤骨科杂志

Objective:To compare the efficacy between arthroscopy-assisted reduction and internal fixation (ARIF) versus open reduction and internal fixation (ORIF) in the treatment of tibial plateau fractures.Methods:A retrospective analysis was done of the 75 patients with tibial plateau fracture who had been treated by ARIF or ORIF at Department of Orthopaedics, Union Hospital Affiliated to Tongji Medical Collage from January 2016 to August 2018. They were 58 men and 17 women, aged from 20 to 54 years (average, 47 years). The left side was affected in 42 cases and the right side in 33. By the Schatzker classification, there were 23 cases of type Ⅰ, 49 cases of type Ⅱ and 3 cases of type Ⅲ. Of them, 40 were treated by ARIF and 35 by ORIF. The 2 groups were compared in terms of operation time, incision length, intraoperative blood loss, hospital stay, postoperative complications and the Hospital for Special Surgery (HSS) scores 12 months after operation.Results:There was no statistically significant difference between the 2 groups in the preoperative general data, showing the 2 groups were comparable ( P>0.05). The patients were followed up for 12 to 15 months (average, 13.5 months) after operation. The wounds in the 75 patients healed at one stage with no complications like neurovascular lesions. All the fractures healed within 6 months after operation. Compared with the ORIF group, the ARIF group had significantly longer operation time (58.3 min ± 4.2 min versus 48.4 min ± 5.2 min), a significantly shorter incision (4.3 cm ± 0.9 cm versus 6.2 cm ± 0.8 cm), and significantly less intraoperative blood loss (60.8 mL ± 4.5 mL versus 72.8 mL ± 6.5 mL) ( P<0.05). There was no significant difference between the 2 groups in hospital stay (5.1 d ± 0.6 d versus 5.5 d ± 1.6 d) ( P>0.05). Fifteen patients in the ARIF group and 5 in the ORIF group were complicated with soft tissue injury, showing a statistically significant difference ( P<0.05). The excellent and good rate by HSS scores was 100% (40/40) for the ARIF group and 85% (34/40) for the ORIF group, showing no significant difference ( P>0.05). Conclusions:In the treatment of tibial plateau fractures of Schatzker types Ⅰ-Ⅲ, both ARIF and ORIF may result in good efficacy. However, ARIF can evaluate and treat the complicated soft tissue injuries to the knee joint more precisely, showing advantages of shorter operation time, a smaller incision and less blood loss.

Effects of m 6A methylase METTL14 mediated long-chain non-coding RNA EIF3J-AS1 on the migration and invasion of cholangiocarcinoma cells / 中华胰腺病杂志

Objective:To investigate the effects of methyltransferase-like protein 14 (METTL14)-mediated long-chain non-coding RNA EIF3J antisense RNA1 (Inc EIF3J-AS1) on the migration and invasion of cholangiocarcinoma cells and its mechanism.Methods:From September 2017 to December 2018, 10 pairs of cholangiocarcinoma and adjacent normal tissues were collected from the First Affiliated Hospital of Naval Medical University, which were surgically resected and pathologically confirmed. The expression of METTL14 mRNA and Inc EIF3J-AS1 in cholangiocarcinoma tissues was detected by fluorescence quantitative PCR, and the protein expression of METTL14 was detected by Western blotting. Cholangiocarcinoma cell lines HUCCTI and RBE were divided into control group and METTL14 or Inc EIF3J-AS1 knockdown group. The corresponding normal lentivirus was transfected in the control group, and METTL14 or Inc EIF3J-AS1 knockdown group was transfected with lentivirus that interfered with the expression of METTL14 or Inc EIF3J-AS1, respectively. Transwell assay was used to detect the ability of cell migration and invasion, and Western blotting was used to detect the expression of epidermal growth factor receptor (EGFR) and AKT protein.Results:The expressions of METTL14 mRNA and lnc EIF3J-AS1 in cholangiocarcinoma tissues were significantly higher than those in adjacent normal tissues (0.075±0.012 vs 0.031±0.006, 0.140±0.032 vs 0.064±0.012), and there was a positive correlation between expression of METTL4 mRNA and expression of lnc EIF3J-AS1 ( r=0.883, P=0.0007). The expression of METTL14 protein in cholangiocarcinoma tissues was higher than that in adjacent normal tissues (0.354±0.131 vs 0.187±0.183). Compared with the control group, the expression of lnc EIF3J-AS1 was significantly lower in METTL14 or Inc EIF3J-AS1 knockdown group (0.217±0.020 vs 1.000±0.052, 0.149±0.066 vs 1.000±0.045). The migration and invasion ability of cell lines HUCCTI and RBE decreased significantly in lnc EIF3J-AS1 knockout group (5.00±0.58 vs 23.33±0.33, 20.33±0.67 vs 70.67±0.33; 12.00±0.58 vs 25.00±2.52, 22.33±0.89 vs 43.67±0.33). The expression of EGFR and p-AKT/AKT protein were also significantly decreased (0.109±0.015 vs 1.000±0.018, 0.226±0.036 vs 1.000±0.051; 0.118±0.052 vs 1.000±0.069, 0.132±0.098 vs 1.000±0.023). The above differences were statistically significant (all P<0.05). Conclusions:Abnormal expression of lnc EIF3J-AS1 in cholangiocarcinoma mediated by METTL14 can promote tumor cell migration and invasion.

Accurately Differentiating COVID-19, Other Viral Infection, and Healthy Individuals Using Multimodal Features via Late Fusion Learning

Effectively identifying COVID-19 patients using non-PCR clinical data is critical for the optimal clinical outcomes. Currently, there is a lack of comprehensive understanding of various biomedical features and appropriate technical approaches to accurately detecting COVID-19 patients. In this study, we recruited 214 confirmed COVID-19 patients in non-severe (NS) and 148 in severe (S) clinical type, 198 non-infected healthy (H) participants and 129 non-COVID viral pneumonia (V) patients. The participants clinical information (23 features), lab testing results (10 features), and thoracic CT scans upon admission were acquired as three input feature modalities. To enable late fusion of multimodality data, we developed a deep learning model to extract a 10-feature high-level representation of the CT scans. Exploratory analyses showed substantial differences of all features among the four classes. Three machine learning models (k-nearest neighbor kNN, random forest RF, and support vector machine SVM) were developed based on the 43 features combined from all three modalities to differentiate four classes (NS, S, V, and H) at once. All three models had high accuracy to differentiate the overall four classes (95.4%-97.7%) and each individual class (90.6%-99.9%). Multimodal features provided substantial performance gain from using any single feature modality. Compared to existing binary classification benchmarks often focusing on single feature modality, this study provided a novel and effective breakthrough for clinical applications. Findings and the analytical workflow can be used as clinical decision support for current COVID-19 and other clinical applications with high-dimensional multimodal biomedical features. One sentence summaryWe trained and validated late fusion deep learning-machine learning models to predict non-severe COVID-19, severe COVID-19, non-COVID viral infection, and healthy classes from clinical, lab testing, and CT scan features extracted from convolutional neural network and achieved predictive accuracy of > 96% to differentiate all four classes at once based on a large dataset of 689 participants.

An Interpretable Machine Learning Framework for Accurate Severe vs Non-severe COVID-19 Clinical Type Classification

Effectively and efficiently diagnosing COVID-19 patients with accurate clinical type is essential to achieve optimal outcomes for the patients as well as reducing the risk of overloading the healthcare system. Currently, severe and non-severe COVID-19 types are differentiated by only a few clinical features, which do not comprehensively characterize complicated pathological, physiological, and immunological responses to SARS-CoV-2 invasion in different types. In this study, we recruited 214 confirmed COVID-19 patients in non-severe and 148 in severe type, from Wuhan, China. The patients comorbidity and symptoms (26 features), and blood biochemistry (26 features) upon admission were acquired as two input modalities. Exploratory analyses demonstrated that these features differed substantially between two clinical types. Machine learning random forest (RF) models using features in each modality were developed and validated to classify COVID-19 clinical types. Using comorbidity/symptom and biochemistry as input independently, RF models achieved >90% and >95% predictive accuracy, respectively. Input features importance based on Gini impurity were further evaluated and top five features from each modality were identified (age, hypertension, cardiovascular disease, gender, diabetes; D-Dimer, hsTNI, neutrophil, IL-6, and LDH). Combining top 10 multimodal features, RF model achieved >99% predictive accuracy. These findings shed light on how the human body reacts to SARS-CoV-2 invasion as a unity and provide insights on effectively evaluating COVID-19 patients severity and developing treatment plans accordingly. We suggest that symptoms and comorbidities can be used as an initial screening tool for triaging, while biochemistry and features combined are applied when accuracy is the priority. One Sentence SummaryWe trained and validated machine learning random forest (RF) models to predict COVID-19 severity based on 26 comorbidity/symptom features and 26 biochemistry features from a cohort of 214 non-severe and 148 severe type COVID-19 patients, identified top features from both feature modalities to differentiate clinical types, and achieved predictive accuracy of >90%, >95%, and >99% when comorbidity/symptom, biochemistry, and combined top features were used as input, respectively.

Epidemiological, Clinical Characteristics and Outcome of Medical Staff Infected with COVID-19 in Wuhan, China: A Retrospective Case Series Analysis

BackgroundsSince December 2019, a novel coronavirus epidemic has emerged in Wuhan city, China and then rapidly spread to other areas. As of 20 Feb 2020, a total of 2,055 medical staff confirmed with coronavirus disease 2019 (COVID-19) caused by SARS-Cov-2 in China had been reported. We sought to explore the epidemiological, clinical characteristics and prognosis of novel coronavirus-infected medical staff. MethodsIn this retrospective study, 64 confirmed cases of novel coronavirus-infected medical staff admitted to Union Hospital, Wuhan between 16 Jan, 2020 to 15 Feb, 2020 were included. Two groups concerned were extracted from the subjects based on duration of symptoms: group 1 ([≤]10 days) and group 2 (>10 days). Epidemiological and clinical data were analyzed and compared across groups. The Kaplan-Meier plot was used to inspect the change in hospital discharge rate. The Cox regression model was utilized to identify factors associated with hospital discharge. FindingsThe median age of medical staff included was 35 years old. 64% were female and 67% were nurses. None had an exposure to Huanan seafood wholesale market or wildlife. A small proportion of the cohort had contact with specimens (5%) as well as patients in fever clinics (8%) and isolation wards (5%). Fever (67%) was the most common symptom, followed by cough (47%) and fatigue (34%). The median time interval between symptoms onset and admission was 8.5 days. On admission, 80% of medical staff showed abnormal IL-6 levels and 34% had lymphocytopenia. Chest CT mainly manifested as bilateral (61%), septal/subpleural (80%) and ground-glass (52%) opacities. During the study period, no patients was transferred to intensive care unit or died, and 34 (53%) had been discharged. Higher body mass index (BMI) ([≥] 24 kg/m2) (HR 0.14; 95% CI 0.03-0.73), fever (HR 0.24; 95% CI 0.09-0.60) and higher levels of IL-6 on admission (HR 0.31; 95% CI 0.11-0.87) were unfavorable factors for discharge. InterpretationIn this study, medical staff infected with COVID-19 have relatively milder symptoms and favorable clinical course, which may be partly due to their medical expertise, younger age and less underlying diseases. Smaller BMI, absence of fever symptoms and normal IL-6 levels on admission are favorable for discharge for medical staff. Further studies should be devoted to identifying the exact patterns of SARS-CoV-2 infection among medical staff.

Development of Novel Biocomposite Scaffold of Chitosan-Gelatin/Nanohydroxyapatite for Potential Bone Tissue Engineering Applications.

In this study, a three-dimensional chitosan-gelatin/nanohydroxyapatite (ChG/nHaP) scaffold was successfully fabricated and characterized in terms of swelling, degradation, cell proliferation, cell attachment, and mineralization characterizations. The ChG/nHaP scaffold was fabricated with a mean pore size of 100-180 µm. Our results showed that the physicochemical and biological properties of the scaffolds were affected by the presence of HaP. The swelling and degradation characteristics of the ChG scaffold were remarkably decreased by the addition of HaP. On the other hand, the presence of HaP remarkably improved the MC3T3-E1 cell attachment and cell growth in the scaffold membrane. The biocompatible nature of the ChG/nHaP scaffold leads to the development of finely scaled mineral deposits on the scaffold membrane. Thus, HaP played an important role in improving the biological performance of the scaffold. Therefore, the ChG/nHaP scaffold could be applied as a suitable material for bone tissue engineering applications.

Sustained release of polyethylene glycol-modified hydrogel-packaged growth factors and endothelial progenitor cells / 中国组织工程研究

BACKGROUND:Many studies have demonstrated that in order to achieve the endothelialization of the polymer materials, surface loading of bioactive factors is an important means and the introduction of human endothelial cells on the surface of the polymer materials wil contribute to increase the biocompatibility of materials. OBJECTIVE:To synthesize the polyethylene glycol hydrogel packaging vascular endothelial growth factor, basic fibroblast growth factor and endothelial progenitor cells, and to observe the sustained release of growth factors and culture status of endothelial progenitor cells. METHODS:The polyethylene glycol-modified hydrogel containing arginine-glycine-aspartic acid peptide was synthesized, and then, rat endothelial progenitor cells, basic fibroblast growth factor, vascular endothelial growth factor, matrix metal oproteinases were successively added. The hydrogel were immersed in PBS, and ELISA was used to test the levels of growth factors in supernatants every 12 hours. Matrix metal oproteinases-2 (100, 1 000 ng) and matrix metal oproteinases-9 (100, 1 000 ng) at different mass were added into the PBS containing hydrogel after 72 hours. ELISA was also used to test the levels of growth factors in supernatants every 12 hours. The hydrogel encapsulating endothelial progenitor cells was cultured in a culture medium for 5 days, and the number of survival cells was counted by a flow cytometer after digestion. RESULTS AND CONCLUSION:Within 12-72 hours, the release percentage of basic fibroblast growth factor and vascular endothelial growth factor remained at about 41%. After 72 hours, the release percentage of both growth factors was found to grow steadily when matrix metal oproteinases-2 (100, 1000 ng) and matrix metal oproteinases-9 (100, 1 000 ng) at different mass were added, which reached 95%. The release percentage was increased with the increasing mass of matrix metal oproteinases. After 5 days of culture, 88.17%cells stil survived in the hydrogel. These findings indicate that sel-assembly polyethylene glycol-modified hydrogel can realize the control ed release of basic fibroblast growth factor and vascular endothelial growth factor, and it can also support the proliferative growth of endothelial progenitor cells.