SIRT1 and ZNF350 as novel biomarkers for osteoporosis: a bioinformatics analysis and experimental validation.

BACKGROUND: Osteoporosis (OP) is characterized by bone mass decrease and bone tissue microarchitectural deterioration in bone tissue. This study identified potential biomarkers for early diagnosis of OP and elucidated the mechanism of OP. METHODS: Gene expression profiles were downloaded from Gene Expression Omnibus (GEO) for the GSE56814 dataset. A gene co-expression network was constructed using weighted gene co-expression network analysis (WGCNA) to identify key modules associated with healthy and OP samples. Functional enrichment analysis was conducted using the R clusterProfiler package for modules to construct the transcriptional regulatory factor networks. We used the "ggpubr" package in R to screen for differentially expressed genes between the two samples. Gene set variation analysis (GSVA) was employed to further validate hub gene expression levels between normal and OP samples using RT-PCR and immunofluorescence to evaluate the potential biological changes in various samples. RESULTS: There was a distinction between the normal and OP conditions based on the preserved significant module. A total of 100 genes with the highest MM scores were considered key genes. Functional enrichment analysis suggested that the top 10 biological processes, cellular component and molecular functions were enriched. The Toll-like receptor signaling pathway, TNF signaling pathway, PI3K-Akt signaling pathway, osteoclast differentiation, JAK-STAT signaling pathway, and chemokine signaling pathway were identified by Kyoto Encyclopedia of Genes and Genomes pathway analysis. SIRT1 and ZNF350 were identified by Wilcoxon algorithm as hub differentially expressed transcriptional regulatory factors that promote OP progression by affecting oxidative phosphorylation, apoptosis, PI3K-Akt-mTOR signaling, and p53 pathway. According to RT-PCR and immunostaining results, SIRT1 and ZNF350 levels were significantly higher in OP samples than in normal samples. CONCLUSION: SIRT1 and ZNF350 are important transcriptional regulatory factors for the pathogenesis of OP and may be novel biomarkers for OP treatment.

Transcriptomics and Metabolomics Unveil the Neuroprotection Mechanism of AnGong NiuHuang (AGNH) Pill Against Ischaemic Stroke Injury.

As a famous prescription in China, AnGong NiuHuang (AGNH) pill exerts good neuroprotection for ischaemic stroke (IS), but its mechanism is still unclear. In this study, the neuroprotection of AGNH was evaluated in the rat IS model which were established with the surgery of middle cerebral artery occlusion (MCAO), and the potential mechanism was elucidated by transcriptomic analysis and metabolomic analysis. AGNH treatment obviously decreased the infarct volume and Zea-Longa 5-point neurological deficit scores, improved the survival percentage of rats, regional cerebral blood flow (rCBF), and rat activity distance and activity time. Transcriptomics showed that AGNH exerted its anti-inflammatory effects by affecting the regulatory network including Tyrobp, Syk, Tlr2, Myd88 and Ccl2 as the core. Integrating transcriptomics and metabolomics identified 8 key metabolites regulated by AGNH, including L-histidine, L-serine, L-alanine, fumaric acid, malic acid, and N-(L-arginino) succinate, 1-pyrroline-4-hydroxy-2-carboxylate and 1-methylhistamine in the rats with IS. Additionally, AGNH obviously reduced Tyrobp, Syk, Tlr2, Myd88 and Ccl2 at both the mRNA and protein levels, decreased IL-1ß, KC-GRO, IL-13, TNF-α, cleaved caspase 3 and p65 nucleus translocation, but increased IκBα expression. Network pharmacology analysis showed that quercetin, beta-sitosterol, baicalein, naringenin, acacetin, berberine and palmatine may play an important role in protecting against IS. Taken together, this study reveals that AGNH reduced neuroinflammation and protected against IS by inhibiting Tyrobp/Syk and Tlr2/Myd88, as well as NF-κB signalling pathway and regulating multiple metabolites.

A two-step neural network-based guiding system for obtaining reliable radiographs for critical shoulder angle measurement.

Background: The critical shoulder angle (CSA) has been reported to be highly associated with rotator cuff tears (RCTs) and an increased risk of RCT re-tears. However, the measurement of the CSA is greatly affected by the malpositioning of the shoulder. To address this issue, a two-step neural network-based guiding system was developed to obtain reliable CSA radiographs, and its feasibility and accuracy was evaluated. Methods: A total of 1,754 shoulder anteroposterior (AP) radiographs were retrospectively acquired to train and validate a two-step neural network-based guiding system to obtain reliable CSA radiographs. The study included patients aged 18 years or older who underwent X-rays and/or computed tomography (CT) scans of the shoulder. Patients who had undergone shoulder surgery, had a confirmed fracture, or were diagnosed with a musculoskeletal tumor or glenoid defect were excluded from the study. The system consisted of a two-step neural network that in the first step, localized the region of interest of the shoulder, and in the second step, classified the radiography according to type [i.e., 'forward' when the non-overlapping coracoid process is above the glenoid rim, 'backward' when the non-overlapping coracoid process is below or aligned with the glenoid rim, a ratio of the transverse to longitudinal diameter of the glenoid projection (RTL) ≤0.25, or a RTL >0.25]. The performance of the model was assessed in an offline, prospective manner, focusing on the sensitivity and specificity for the forward, backward, RTL ≤0.25, or RTL >0.25 types (denoted as SensF, B, -, + and SpecF, B, -, +, respectively), and Cohen's kappa was also reported. Results: Of 273 cases in the offline prospective test, the SensF, SensB, Sens-, and Sens+ were 88.88% [95% confidence interval (CI): 50.67-99.41%], 94.11% (95% CI: 82.77-98.47%), 96.96% (95% CI: 91.94-99.02%), and 95.06% (95% CI: 87.15-98.40%), respectively. The SpecF, SpecB, Spec-, and Spec+ were 98.48% (95% CI: 95.90-99.51%), 99.55% (95% CI: 97.12-99.97%), 95.04% (95% CI: 89.65-97.81%), and 97.39% (93.69-99.03%), respectively. A high classification rate (93.41%; 95% CI: 89.14-96.24%) and almost perfect agreement (Cohen's kappa: 0.903, 95% CI: 0.86-0.95) were achieved. Conclusions: The guiding system can rapidly and accurately classify the types of AP shoulder radiography, thereby guiding the adjustment of patient positioning. This will facilitate the rapid obtainment of reliable CSA radiography to measure the CSA on proper AP radiographs.

Local Injection of Rapamycin-Loaded Pcl-Peg Nanoparticles for Enhanced Tendon Healing in Rotator Cuff Tears via Simultaneously Reducing Fatty Infiltration and Drug Toxicity.

As a common cause of shoulder pain, rotator cuff tears (RCTs) are difficult to treat clinically because of their unsatisfactory prognosis due to the fatty infiltration caused by muscle-derived stem cells (MDSCs). Previous studies have found that rapamycin (RAPA) can inhibit fatty infiltration. However, systemic administration of RAPA may cause complications such as infection and nausea, while local administration of RAPA may lead to the cytotoxicity of tendon cells, affecting the healing of rotator cuffs. In this study, biocompatible and clinically approved polycaprolactone-polyethylene glycol (PCL-PEG) is formulated into an injectable nanoparticle for the sustained release of RAPA. The results indicate that the RAPA/PCL-PEG nanoparticles (NPs) can efficiently prolong the release of RAPA and significantly reduce the cytotoxicity of tendon cells caused by RAPA. The study of the fatty infiltration model in rats with delayed rotator cuff repair shows that weekly intraarticular injection of RAPA/PCL-PEG NPs can more effectively reduce the fatty infiltration and muscle atrophy of rat rotator cuffs and leads to better mechanical properties and gait improvements than a daily intraarticular injection of RAPA. These findings imply that local injection of RAPA/PCL-PEG NPs in the shoulder joints can be a potential clinical option for RCTs patients with fatty infiltration.

Modified Continuous Arthroscopy-Guided Suprascapular Nerve Block for Postoperative Pain Control Following Rotator Cuff Repair: Surgical Technique.

Arthroscopic rotator cuff repair is one of the most painful surgical procedures; patients complain of pain especially during the first 48 hours postoperatively. Pain management is an essential goal to reduce the demand for analgesic agents and patient discomfort. Various methods have been introduced for arthroscopic rotator cuff repair, including continuous arthroscopy-guided suprascapular nerve blocks (SSNB) and interscalene nerve blocks. However, the aforementioned procedures have shown disadvantages such as catheter mobilization, which may cause injury to the artery, a weak analgesic effect, and Horner syndrome, as well as phrenic nerve paralysis. A modification of the continuous arthroscopy-guided SSNB has been introduced at our hospital: the modified continuous arthroscopy-guided SSNB technique. The aim of this technique is to immobilize the catheter to reduce the chance of injury and minimize postsurgical analgesic needs and patient discomfort.

Enhancing prediction of supraspinatus/infraspinatus tendon complex injuries through integration of deep visual features and clinical information: a multicenter two-round assessment study.

OBJECTIVE: Develop and evaluate an ensemble clinical machine learning-deep learning (CML-DL) model integrating deep visual features and clinical data to improve the prediction of supraspinatus/infraspinatus tendon complex (SITC) injuries. METHODS: Patients with suspected SITC injuries were retrospectively recruited from two hospitals, with clinical data and shoulder x-ray radiographs collected. An ensemble CML-DL model was developed for diagnosing normal or insignificant rotator cuff abnormality (NIRCA) and significant rotator cuff tear (SRCT). All patients suspected with SRCT were confirmed by arthroscopy examination. The model's performance was evaluated using sensitivity, specificity, accuracy, and area under the curve (AUC) metrics, and a two-round assessment was conducted to authenticate its clinical applicability. RESULTS: A total of 974 patients were divided into three cohorts: the training cohort (n = 828), the internal validation cohort (n = 89), and the external validation cohort (n = 57). The CML-DL model, which integrates clinical and deep visual features, demonstrated superior performance compared to individual models of either type. The model's sensitivity, specificity, accuracy, and area under curve (95% confidence interval) were 0.880, 0.812, 0.836, and 0.902 (0.858-0.947), respectively. The CML-DL model exhibited higher sensitivity and specificity compared to or on par with the physicians in all validation cohorts. Furthermore, the assistance of the ensemble CML-DL model resulted in a significant improvement in sensitivity for junior physicians in all validation cohorts, without any reduction in specificity. CONCLUSIONS: The ensembled CML-DL model provides a solution to help physicians improve the diagnosis performance of SITC injury, especially for junior physicians with limited expertise. CRITICAL RELEVANCE STATEMENT: The ensembled clinical machine learning-deep learning (CML-DL) model integrating deep visual features and clinical data provides a superior performance in the diagnosis of supraspinatus/infraspinatus tendon complex (SITC) injuries, particularly for junior physicians with limited expertise. KEY POINTS: 1. Integrating clinical and deep visual features improves diagnosing SITC injuries. 2. Ensemble CML-DL model validated for clinical use in two-round assessment. 3. Ensemble model boosts sensitivity in SITC injury diagnosis for junior physicians.

Molecular and optical signatures of photochemical transformation of dissolved organic matter: Nonnegligible role of suspended particulate matter in urban river.

Natural dissolved organic matter (DOM) is one of the Earth's dynamic carbon pools and a key intermediate in the global carbon cycle. Photochemical processes potentially affect DOM composition and activity in surface water. Suspended particulate matter (SPM) is the integral component of slow-moving rivers, and holds the potential for photochemical reactivity. To further investigate the influence of SPM on DOM photochemical transformation, this study conducted experiments comparing samples with and without SPM irradiated under simulated sunlight. Surface water samples from slow-moving urban rivers were collected. DOM optical characteristics and molecular features obtained by Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS) were investigated. Photolabile DOM was enriched in unsaturated and highly aromatic terrestrial substances. Photoproduced DOM had low aromaticity and was dominated by saturated aliphatics, protein-like substances, and carbohydrates. Study results indicated that the presence of SPM had a nonnegligible impact on the molecular traits of DOM, such as composition, molecular diversity, photolability, and bioavailability during photochemical reactions. In the environment affected by SPM, molecules containing heteroatoms exhibit higher photosensitivity. SPM promotes the photochemical transformation of a wider range of chemical types of photolabile DOM, particularly nitrogen-containing compounds. This study provides an essential insight into the more precise simulation of photochemical reactions of DOM influenced by SPM occurring in natural rivers, contributing to our understanding of the global carbon cycle from new theoretical perspectives.

Deep learning for automated measurement of CSA related acromion morphological parameters on anteroposterior radiographs.

BACKGROUND: The Critical Shoulder Angle Related Acromion Morphological Parameter (CSA- RAMP) is a valuable tool in the analyzing the etiology of the rotator cuff tears (RCTs). However, its clinical application has been limited by the time-consuming and prone to inter- and intra-user variability of the measurement process. OBJECTIVES: To develop and validate a deep learning algorithm for fully automated assessment of shoulder anteroposterior radiographs associated with RCTs and calculation of CSA-RAMP. METHODS: Retrospective analysis was conducted on radiographs obtained from computed tomography (CT) scans and X-rays performed between 2018 and 2020 at our institution. The development of the system involved the utilization of digitally reconstructed radiographs (DRRs) generated from each CT scan. The system's performance was evaluated by comparing it with manual and semiautomated measurements on two separate test datasets: dataset I (DRRs) and dataset II (X-rays). Standard metrics, including mean average precision (AP), were utilized to assess the segmentation performance. Additionally, the consistency among fully automated, semiautomated, and manual measurements was comprehensively evaluated using the Pearson correlation coefficient and Bland-Altman analysis. RESULTS: A total of 1080 DRRs generated from 120 consecutive CT scans and 159 X-ray films were included in the study. The algorithm demonstrated excellent segmentation performance, with a mean AP of 57.67 and an AP50 of 94.31. Strong inter-group correlations were observed for all CSA-RAMP measurements in both test datasets I (automated versus manual, automated versus semiautomated, and semiautomated versus manual; r = [0.990---0.997], P < 0.001) and dataset II (r = [0.984---0.995], P < 0.001). Bland-Altman analysis revealed low bias for all CSA-RAMP measurements in both test datasets I and II, except for CD (with a maximum bias of 2.49%). CONCLUSIONS: We have successfully developed a fully automated algorithm capable of rapidly and accurately measuring CSA-RAMP on shoulder anteroposterior radiographs. A consistent automated CSA- RAMP measurement system may accelerate powerful and precise studies of disease biology in future large cohorts of RCTs patients.

LARAI portal provides a safe method for lateral meniscus repair: three-dimensional computed tomography and cadaveric assessment.

BACKGROUND: Lateral, All-Round and All-Inside (LARAI) portal is a viewing or working portal for observing and repairing the lesions of the lateral meniscus. However, there are safety concerns about popliteal artery (PA) injuries during the procedure. This study aimed to assess the safe distance between the trajectory of the LARAI portal and PA. MATERIALS AND METHODS: Both three-dimensional computed tomography (3D-CT) and cadavers were used to simulate the LARAI portal trajectory. In the 3D-CT study, between January 2020 and September 2020, 45 participants who underwent computed tomography angiography were included in the study. The shortest distance from the PA to the simulated trajectory needle (PS) was measured using 3D-CT. Mean -3SD -2 was calculated to assess the safety of the LARAI portal trajectory. If this value was more than zero, the trajectory was considered "safe." In the cadaveric study, lower limbs from seven fresh-frozen cadavers were used to establish the "safe" trajectories of the LARAI portal, and the PS was measured. RESULTS: In the 3D-CT study, the longest PS (P < 0.001) was found 20 mm lateral to the edge of the patellar tendon trajectory at 0 mm from the posterior cruciate ligament (PCL). Safe trajectories were also found 10 mm, 15 mm, and 20 mm lateral to the edge of the patellar tendon at 0 mm from the PCL, as well as the 20 mm lateral to the edge of the patellar tendon at 3 mm from the PCL. The cadaveric study showed that the average PS of all safe trajectories closely adjoined to PCL was greater than 14 mm. CONCLUSIONS: The LARAI portal trajectory in the "figure of four" is safe, and the optimal insertion point is 10-20 mm lateral to the edge of the patellar tendon and closely adjoined to the posterolateral margin of the PCL at knee joint line level. LEVEL OF EVIDENCE: Level IV.

Salvianolic acid B attenuates inflammation and prevent pathologic fibrosis by inhibiting CD36-mediated activation of the PI3K-Akt signaling pathway in frozen shoulder.

Frozen shoulder (FS) is characterized by pain and limited range of motion (ROM). Inflammation and fibrosis are accepted as main pathologic processes associated with the development of FS. However, the intrinsic mechanisms underlying pathologic fibrosis remain unclear. We aimed to elucidate the key molecules involved in pathologic fibrosis and explore new therapeutic targets for FS. Synovial fibroblasts isolated from patient biopsies were identified using immunofluorescence. Western blotting, RT-qPCR, cell adhesion tests, and would-healing assays were used to evaluate the fibrosis-related functions of synovial fibroblasts. Elevated cluster of differentiation 36 (CD36) expression was detected in FS using Western blotting and immunohistochemistry. Salvianolic acid b (SaB) inhibited CD36, blocking synovial fibroblast-induced inflammation and fibrosis. Our RNA-seq data showed that knocking down CD36 dramatically impaired the capacity of synovial fibroblasts for cell adhesion and that the PI3K-Akt signaling pathway may be crucial to the fibrotic process of FS. By up-regulating CD36 and inhibiting the phosphorylation of Akt, we demonstrated that CD36 promotes pathologic fibrosis by activating the PI3k-Akt pathway. Finally, rats treated with SaB had improved ROM and less collagen fiber deposition than the FS model group. Conclusion: SaB attenuates inflammation and inhibited the CD36-mediated activation of the PI3K-Akt signaling pathway to block pathologic fibrosis of FS in vitro and in vivo models.

[Effect of critical shoulder angle on deltoid muscle strength reduction in patients with rotator cuff tears].

Objective: To investigate the synergistic interaction between the deltoid muscle and the rotator cuff muscle group in patients with rotator cuff tears (RCT), as well as the impact of the critical shoulder angle (CSA) on deltoid muscle strength. Methods: A retrospective analysis was conducted on clinical data from 42 RCT patients who met the selection criteria and were treated between March 2022 and March 2023. There were 13 males and 29 females, with an age range of 42-77 years (mean, 60.5 years). Preoperative visual analogue scale (VAS) score was 6.0±1.6. CSA measurements were obtained from standard anteroposterior X-ray films before operation, and patients were divided into two groups based on CSA measurements: CSA>35° group (group A) and CSA≤35° group (group B). Handheld dynamometry was used to measure the muscle strength of various muscle group in the shoulder (including the supraspinatus, infraspinatus, subscapularis, and anterior, middle, and posterior bundles of the deltoid). The muscle strength of the unaffected side was compared to the affected side, and muscle imbalance indices were calculated. Muscle imbalance indices between male and female patients, dominant and non-dominant sides, and groups A and B were compared. Pearson correlation analysis was used to examine the relationship between muscle imbalance indices and CSA as well as VAS scores. Results: Muscle strength in all muscle groups on the affected side was significantly lower than on the unaffected side ( P<0.05). The muscle imbalance indices for the supraspinatus, subscapularis, infraspinatus, and anterior, middle, and posterior bundles of the deltoid were 14.8%±24.4%, 5.9%±9.7%, 7.2% (0, 9.1%), 17.2% (5.9%, 26.9%), 8.3%±21.3%, and 10.2% (2.8%, 15.4%), respectively. The muscle imbalance indices of the anterior bundle of the deltoid, supraspinatus, and infraspinatus were significantly lower in male patients compared to female patients ( P<0.05); however, there was no significant difference in muscle imbalance indices among other muscle groups between male and female patients or between the dominant and non-dominant sides ( P>0.05). There was a positive correlation between the muscle imbalance indices of infraspinatus and VAS score ( P<0.05), and a positive correlation between CSA and the muscle imbalance indices of middle bundle of deltoid ( P<0.05). There was no correlation between the muscle imbalance indices of other muscle groups and VAS score or CSA ( P>0.05). Preoperative CSA ranged from 17.6° to 39.4°, with a mean of 31.1°. There were 9 cases in group A and 33 cases in group B. The muscle imbalance indices of the anterior bundle of the deltoid was significantly lower in group A compared to group B ( P<0.05), while there was no significant difference in muscle imbalance indices among other muscle groups between group A and group B ( P>0.05). Conclusion: Patients with RCT have a phenomenon of deltoid muscle strength reduction, which is more pronounced in the population with a larger CSA.

Single-cell biology: what does the future hold?

In this Editorial, our Chief Editor and members of our Advisory Editorial Board discuss recent breakthroughs, current challenges, and emerging opportunities in single-cell biology and share their vision of "where the field is headed."

The Longitude-Latitude-Loop Used for Complex Bankart Lesion Repair: An All-Arthroscopic Technique.

The most frequent operation for anterior shoulder instability is the arthroscopic Bankart repair, which has a positive outcome and a low rate of complications. A variety of restoration procedures have been reported to reconstruct labral height and reproduce a dynamic concavity-compression reaction. The longitude-latitude loop is a knotless high-strength suture method that simultaneously tightens the joint capsule in the warp and weft direction and resists tearing. The suture method is safe and reproducible. This study aimed to present a longitude-latitude loop suture for joint capsule labral complex repair during Bankart arthroscopy surgery.

Hematologic side effects of immune checkpoint inhibitor with or without chemotherapy in patients with advanced and metastatic gastrointestinal cancer: A systematic review and network meta-analysis of phase 3 trials.

Background: The regimens of immune checkpoint inhibitors (ICIs) alone or with chemotherapy are emerging as systemic therapy for patients with advanced and metastatic gastrointestinal cancers. However, the risk of treatment-related hematologic toxicity stays unclear. Methods: We enrolled in phase 3 randomized clinical trials (RCTs) comparing PD-1, PD-L1, and CTLA-4 inhibitors in advanced and metastatic gastrointestinal cancers. The incidences of overall treatment-related adverse events (TRAEs), discontinuation, leukopenia, neutropenia, thrombocytopenia, and anemia were extracted for the Bayesian network meta-analysis. Analyses with poor convergence or low incidence were reported as incidences with 95% CIs instead. Results: Sixteen phase 3 RCTs with 9732 patients who received systemic therapy were included. A total of 150 (1.54% [95% CI 1.31-1.80]) treatment-related death events were recorded, whereas 13 (0.13% [95% CI 0.08-0.22]) of them were hematologic. 0.24% (95% CI 0.12-0.48) patients received ICI plus chemotherapy were recorded for hematological deaths, 0.09% (95% CI 0.01-0.23) were for chemotherapy alone, and 0.05% were for ICI alone (95% CI 0.01-0.29). Febrile neutropenia was the most frequent cause of death in ICI with chemotherapy. For grade ≥3 TRAEs, we found nivolumab plus chemotherapy (OR 1.63 [95% CI 0.84-3.17]) had a higher risk than other treatments. Overall, ICI monotherapy led to fewer AEs than chemotherapy-based regimens in the analyses of leukopenia, neutropenia, thrombocytopenia, and anemia. Among the 11 treatments, toripalimab plus chemotherapy possessed the highest risk in any-grade leukopenia (OR 1.84 [95% CI 0.48, 6.82]) and neutropenia (OR 1.71 [95% CI 0.17, 17.40]) respectively. For grade ≥3 hematologic AEs, neutropenia (20.08% [95% CI 18.67-21.56]) related to ICI plus chemotherapy was the most dominant. ICI plus chemotherapy was likely to increase the incidence than dosing these drugs alone. Conclusion: Using ICI alone had a low incidence of causing hematologic mortality and AEs, while the combination with chemotherapy might magnify the side effects. Comprehensively, pembrolizumab plus chemotherapy and sintilimab plus chemotherapy were the safest regimens in terms of leukopenia and neutropenia respectively. This study will guide clinical practice for ICI-based chemotherapy. Systematic Review Registration: PROSPERO, identifier CRD42022380150.

Multi-omics reveals Dengzhan Shengmai formulation ameliorates cognitive impairments in D-galactose-induced aging mouse model by regulating CXCL12/CXCR4 and gut microbiota.

Dengzhan Shengmai (DZSM), a traditional Chinese medicine formulation, has been administered extensively to elderly individuals with cognitive impairment (CI). However, the underlying mechanisms by which Dengzhan Shengmai improves cognitive impairment remains unknown. This study aimed to elucidate the underlying mechanism of the effect of Dengzhan Shengmai on aging-associated cognitive impairment via a comprehensive combination of transcriptomics and microbiota assessment. Dengzhan Shengmai was orally administered to a D-galactose-induced aging mouse model, and evaluation with an open field task (OFT), Morris water maze (MWM), and histopathological staining was performed. Transcriptomics and 16S rDNA sequencing were applied to elucidate the mechanism of Dengzhan Shengmai in alleviating cognitive deficits, and enzyme-linked immunosorbent assay (ELISA), quantitative real-time polymerase chain reaction (PCR), and immunofluorescence were employed to verify the results. The results first confirmed the therapeutic effects of Dengzhan Shengmai against cognitive defects; specifically, Dengzhan Shengmai improved learning and impairment, suppressed neuro loss, and increased Nissl body morphology repair. Comprehensive integrated transcriptomics and microbiota analysis indicated that chemokine CXC motif receptor 4 (CXCR4) and its ligand CXC chemokine ligand 12 (CXCL12) were targets for improving cognitive impairments with Dengzhan Shengmai and also indirectly suppressed the intestinal flora composition. Furthermore, in vivo results confirmed that Dengzhan Shengmai suppressed the expression of CXC motif receptor 4, CXC chemokine ligand 12, and inflammatory cytokines. This suggested that Dengzhan Shengmai inhibited CXC chemokine ligand 12/CXC motif receptor 4 expression and modulated intestinal microbiome composition by influencing inflammatory factors. Thus, Dengzhan Shengmai improves aging-related cognitive impairment effects via decreased CXC chemokine ligand 12/CXC motif receptor 4 and inflammatory factor modulation to improve gut microbiota composition.

Single-cell RNA sequencing reveals that VIM and IFITM3 are vital targets of Dengzhan Shengmai capsule to protect against cerebral ischemic injury.

ETHNOPHARMACOLOGICAL RELEVANCE: Ischemic stroke is one of the leading causes of mortality, but therapies are limited. Dengzhan Shengmai capsule (DZSM) was included by the Chinese Pharmacopoeia 2020 and has been broadly used for the treatment of ischemic stroke. However, the mechanism of DZSM against ischemic stroke is unclear. AIM OF THE STUDY: This study used RNA sequencing (RNA-seq) and single-cell RNA sequencing (scRNA-seq) to investigate the mechanism of action of DZSM against ischemic stroke. MATERIALS AND METHODS: The rats were randomly divided into six groups: the Sham, I/R (water), I/R + DZSM-L (0.1134g/kg), I/R + DZSM-H (0.4536g/kg), I/R + NMDP (20mg/kg), and I/R + Ginaton (20mg/kg). The rats were administrated drugs for 5 days then followed by the ischemic brain injury caused by middle cerebral artery occlusion (MCAO). The neuroprotective effect was assessed by infraction rate, neurological deficit scores, regional cerebral blood flow (rCBF), hematoxylin and eosin (H&E) staining, and Nissl staining. Based on RNA-seq and scRNA-seq, the vital biological processes and core targets of DZSM against cerebral ischemia were revealed. Enzyme-linked immunosorbent assay (ELISA) and immunofluorescence (IF) staining were used to investigate the vital biological processes and core targets of DZSM against ischemic stroke. RESULTS: Administration of DZSM significantly reduced the infarction rate and Zea Longa score, Garcia JH score, and ameliorated the reduction in rCBF. And alleviated the neuronal damage, such as increased neuronal density level and Nissl bodies density level. RNA-seq analysis revealed that DZSM played important roles in inflammation and apoptosis. ELISA and IF straining validation confirmed that DZSM significantly decreased the expression of IL-6, IL-1ß, TNF-α, ICAM-1, IBA-1, MMP9, and Cleaved caspase-3 in MCAO rats. ScRNA-seq analysis identified 8 core targets in neurons including HSPB1, SPP1, MT2A, GFAP, IFITM3, VIM, CRIP1, and GPD1, and VIM and IFITM3 was verified to be decreased by DZSM in neurons. CONCLUSION: Our study illustrates the neuroprotective effect of DZSM against ischemia stroke, and VIM and IFITM3 were identified as vital targets in neurons of DZSM in protecting against MCAO-induced I/R injury.

Arthroscopic Autologous Coracoacromial Ligament Augment Technique for Particle Thickness of Rotator Cuff with at Least Two Years Follow-Up.

OBJECTIVE: Patch technology has been the new technique in the treatment of partial thickness of the rotator cuff tear (PTRCTs) to address the limitation of traditional techniques. Compared with allogeneic patches and artificial materials, coracoacromial ligament is obviously closer to their own biology. The purpose of the study was to evaluate the functional and radiographic outcomes following arthroscopic autologous coracoacromial ligament augment technique for treatment of PTRCTs. METHOD: This study included three female patients with PTRCTs who underwent arthroscopy operation in 2017 with an average age of 51 years (range from 50 to 52 years). The coracoacromial ligament implant was attached to the bursal side surface of the tendon. The clinical results were evaluated by American Shoulder and Elbow Surgeons (ASES) score, Simple Shoulder Test (SST), acromiohumeral distance (AHD) and muscle strength before and 12 months after operation. Magnetic resonance imaging (MRI) was performed 24 months after operation to assess the integrity of the anatomical structure of the original tear site. RESULT: The average ASES score improved significantly from 57.3 preoperatively to 95.0 at 1-year follow-up. The strength improved significantly from grade 3 preoperatively to grade 5 at 1 year. Two of three patients underwent the MRI at 2-year follow-up. Radiographic released the complete healing of rotator cuff tear. No implant-related serious adverse events were reported. CONCLUSION: The new technique of using autogenous coracoacromial ligament patch augment provides good clinical results on patients with PTRCTs.

Effective endodontically treated incisors with external root resorption during orthodontic movement: A case report.

This case report describes a 21-year-old orthodontic patient experienced the external apical root resorption of maxillary central incisors with pulpitis during the orthodontic movement. The active cooperation of orthodontists and endodontists demonstrated the satisfactory treatment outcome and prevented further apical root resorption. The etiology of external apical root resorption is comprehensive, orthodontists should be armed with an adequate training and scientific knowledge, and keep the treatment mechanism simple and precise to guard against it. Besides, we should know the right timing of endodontic treatment and applying orthodontic force when external apical root resorption occurs.

Biomechanical Analysis of Lark-Loop, Lasso-Loop and Krackow Suture Technique in Tenodesis.

OBJECTIVE: Strong tendon grasping is vital to the success of a tenodesis operation. The purpose of this study was to evaluate the initial tendon-fixation strength of the Lark-Loop technique in arthroscopic suprapectoral biceps tenodesis and compare it with others commonly used techniques. METHODS: Thirty-three porcine superficial flexor digitorum tendons were harvested from a local slaughterhouse and randomly divided into three groups to perform three tendon fixation techniques (Lasso-Loop stich group, Lark-Loop stich group or Krackow stich group; 11 tendons each group) with a No. 2 suture, respectively. Each tendon was pre-tensioned in 5°N for 2 min and then cyclically loaded 5 to 30°N for 500 cycles to assess displacement. After cyclic loading, the tendon was loaded to ultimate tendon-suture configuration failure at the rate of 1 mm/s. Finally, the mode of failure and the construct stiffness of the tendon were recorded and calculated. RESULTS: After cyclical loading, the displacement of the Lark-Loop group was equivalent to the Krakow group (P > 0.9999) but significantly smaller than the Lasso-Loop group (P = 0.0009). The ultimate load to failure for the Lark-Loop was equivalent to the Krakow technique group (P = 0.1463) but significantly greater than the Lasso-Loop group (P < 0.0001). The stiffness for the Lark-Loop was equivalent to the Krakow group (P = 0.4718) but significantly greater than the Lasso-Loop technique group (P < 0.0001). In the Lark-Loop and Krackow group, all the tendons failed by suture breakage, while all the tendons failed by suture cutting through the tendon in the Lasso-Loop technique group. CONCLUSION: Lark-Loop suture technique has biomechanical properties comparable to Krackow and superior to the Lasso-Loop in terms of suture displacement, ultimate load to failure, and stiffness. Therefore, the Lark-Loop suture fixation technique may be beneficial for arthroscopic biceps tenodesis.

Machine learning model successfully identifies important clinical features for predicting outpatients with rotator cuff tears.

PURPOSE: The aim of this study is to develop a machine learning model to identify important clinical features related to rotator cuff tears (RCTs) using explainable artificial intelligence (XAI) for efficiently predicting outpatients with RCTs. METHODS: A retrospective review of a local clinical registry dataset was performed to include patients with shoulder pain and dysfunction who underwent questionnaires and physical examinations between 2019 and 2022. RCTs were diagnosed by shoulder arthroscopy. Six machine-learning algorithms (Stacking, Gradient Boosting Machine, Bagging, Random Forest, Extreme Gradient Boost (XGBoost), and Adaptive Boosting) were developed for the prediction. The performance of the models was assessed by the area under the receiver operating characteristic curve (AUC), Brier scores, and Decision curve. The interpretability of the predicted outcomes was evaluated using Shapley additive explanation (SHAP) values. RESULTS: A total of 1684 patients who completed questionnaires and clinical tests were included, and 417 patients with RCTs underwent shoulder arthroscopy. In six machining learning algorithms for predicting RCTs, the accuracy, AUC values, and Brier scores were in the range of 0.81-0.86, 0.75-0.92, and 0.15-0.19, respectively. The XGBoost model showed superior performance with accuracy, AUC, and Brier scores of 0.85(95% confidence interval, 0.82-0.87), 0.92 (95% confidence interval,0.90-0.94), and 0.15 (95% confidence interval,0.14-0.16), respectively. The Shapley plot showed the impact of the clinical features on predicting RCTs. The most important variables were Jobe test, Bear hug test, and age for prediction, with mean SHAP values of 1.458, 0.950, and 0.790, respectively. CONCLUSION: The machine learning model successfully identified important clinical variables for predicting patients with RCTs. In addition, the best algorithm was also integrated into a digital application to provide predictions in outpatient settings. This tool may assist patients in reducing their pain experience and providing prompt treatments. LEVEL OF EVIDENCE: Level III.