Linear polyubiquitylation of Gli protein regulates its protein stability and facilitates tumor growth in colorectal cancer.

The linear ubiquitin chain assembly complex (LUBAC) mediates the linear ubiquitination of various proteins and is involved in NF-κB signaling and immune regulation. However, the function and mechanism of linear ubiquitination in regulating oncogenic signaling and tumor growth have remained poorly understood. Herein, we identified Gli proteins, key transcription factors in the Hedgehog (Hh) signaling pathway, as novel substrates of LUBAC. Linear ubiquitination stabilizes Gli proteins, leading to the noncanonical activation of Hh signaling in CRC cells. Furthermore, LUBAC facilitates tumor growth in CRC cells. Additionally, elevated expression of LUBAC components in CRC tissues was observed, and higher expression levels of these components correlated with poor prognosis in CRC patients. Interestingly, inhibition of LUBAC using either a small molecule agonist or RNA silencing specifically suppressed cell growth in CRC cells but had no effect on normal intestinal cells. Taken together, aberrant expression of LUBAC components activates Hh signaling noncanonically by mediating linear ubiquitination, promoting tumor growth in CRC, demonstrating the novel function of linear ubiquitination in regulating the protein stability of its substrates and highlighting the potential of targeting LUBAC as a therapeutic strategy in CRC.

Robust Nuclear Spin Polarization via Ground-State Level Anticrossing of Boron Vacancy Defects in Hexagonal Boron Nitride.

Nuclear spin polarization plays a crucial role in quantum information processing and quantum sensing. In this work, we demonstrate a robust and efficient method for nuclear spin polarization with boron vacancy (V_{B}^{-}) defects in hexagonal boron nitride (h-BN) using ground-state level anticrossing (GSLAC). We show that GSLAC-assisted nuclear polarization can be achieved with significantly lower laser power than excited-state level anticrossing, making the process experimentally more viable. Furthermore, we have demonstrated direct optical readout of nuclear spins for V_{B}^{-} in h-BN. Our findings suggest that GSLAC is a promising technique for the precise control and manipulation of nuclear spins in V_{B}^{-} defects in h-BN.

Interspinous Motion Measurement Could Serve as a Quantitative Method for Assessing Bony Fusion after Anterior Cervical Corpectomy and Fusion (ACCF).

STUDY DESIGN: A retrospective diagnostic test study. OBJECTIVE: To establish quantitative fusion criteria for ACCF. SUMMARY OF BACKGROUND DATA: Currently, the criteria of fusion after ACCF remains controversial. Evaluation techniques such as observation of continuous bony trabeculae across the fusion site, were qualitative and conducted by naked eyes, limiting its accuracy. Dynamic fusion indicators such as interspinous motion (ISM), have been successfully utilized in anterior cervical discectomy and fusion (ACDF) for evaluating fusion and detecting pseudarthrosis. However, the accuracy of ISM in ACCF is unclear. METHODS: Patients underwent ACCF in our hospital were reviewed. Imaging data including dynamic radiographs and computed tomography (CT) scans were collected. The distance and angle form of ISM (ISM-dis and ISM-ang) were measured on dynamic radiograph, with CT as the gold standard for fusion. The correlation between ISM methods was analyzed using Spearmann correlation. Receiver Operating Characteristic (ROC) curves were used to evaluate the accuracy and generate optimal cut-off values. Three observers independently measured the data twice to determine interobserver and intraobserver reliability. RESULTS: Our study included 178 imaging datasets. ISM-dis showed positive correlation with ISM-ang (r=0.914). The area under ROC (AUC) for ISM-dis and ISM-ang was 0.926 and 0.893. Applying a cutoff value of 1.50 mm to ISM-dis yielded a sensitivity of 87.8% and specificity of 92.3% for detecting pseudarthrosis. For ISM-ang, a cutoff value of 1.75° resulted in a sensitivity of 79.1% and specificity of 92.3%. Intraobserver reliabilities for ISM-dis (0.986, 0.984 and 0.972) were higher than for ISM-ang (0.935, 0.963 and 0.935), as was interobserver reliability (0.985 for ISM-dis and 0.956 for ISM-ang). Fusion rates calculated using ISM-dis and ISM-ang were 52.4% and 61.9%. CONCLUSION: ISM can serve as an alternative to CT for assessing fusion, with ISM-dis at a 1.50 mm cutoff and ISM-ang at 1.75° demonstrating high diagnostic accuracy.

Suppressing Thermal Noise to Sub-Millikelvin Level in a Single-Spin Quantum System Using Realtime Frequency Tracking.

A single nitrogen-vacancy (NV) center in a diamond can be used as a nanoscale sensor for magnetic field, electric field or nuclear spins. Due to its low photon detection efficiency, such sensing processes often take a long time, suffering from an electron spin resonance (ESR) frequency fluctuation induced by the time-varying thermal perturbations noise. Thus, suppressing the thermal noise is the fundamental way to enhance single-sensor performance, which is typically achieved by utilizing a thermal control protocol with a complicated and highly costly apparatus if a millikelvin-level stabilization is required. Here, we analyze the real-time thermal drift and utilize an active way to alternately track the single-spin ESR frequency drift in the experiment. Using this method, we achieve a temperature stabilization effect equivalent to sub-millikelvin (0.8 mK) level with no extra environmental thermal control, and the spin-state readout contrast is significantly improved in long-lasting experiments. This method holds broad applicability for NV-based single-spin experiments and harbors the potential for prospective expansion into diverse nanoscale quantum sensing domains.

Comparison of Postoperative Pain and Surgical Outcomes Between Three Types of Modified Muscle-Sparing Laminoplasty and Conventional Laminoplasty for Multilevel Degenerative Cervical Myelopathy.

STUDY DESIGN: Retrospective cohort study. OBJECTIVE: This study aimed to compare postoperative pain and surgical outcomes of open-door laminoplasty (LP) and three types of muscle-sparing laminoplasties, namely unilateral muscle-preservation laminoplasty (UL), spinous process splitting double-door laminoplasty (DL) and intermuscular "raising roof" laminoplasty (RL) for multilevel degenerative cervical myelopathy (MDCM). METHODS: Consecutive MDCM patients underwent LP or modified laminoplasties (UL, DL, RL) in 2022 were enrolled. Patients' preoperative baseline data and surgical characteristics were collected. Postoperative transient pain (TP), the axial pain and Japanese Orthopedic Association (JOA) score and neck disability index (NDI) at 6-month and 12-month follow-up were documented. RESULTS: A total of 154 MDCM patients were included and a 12-month follow-up was completed for 148 patients (LP: 36, UL:39, DL: 37, RL:36). No significant difference was observed in the baseline data. Four groups presented favorable and comparable surgical outcome. The RL group reported significantly the least severe TP on the first three days following surgery. However, no significant difference was found in the axial pain and axial symptoms at both follow-ups. After regression analysis, RL group exhibited significantly better efficacy in alleviating Day-1 TP (P = 0.047) and 6-month axial pain (P = 0.040). However, this superiority was not observed at 12-month follow-up. CONCLUSION: All the three muscle-sparing laminoplasty procedures showed similar short-term surgical outcomes compared to LP. The RL procedure demonstrated superiority in alleviating TP and 6-month axial pain compared to LP. The RL and DL groups showed less C5 palsy compared to LP.

The safety, immunogenicity, and efficacy of heterologous boosting with a SARS-CoV-2 mRNA vaccine (SYS6006) in Chinese participants aged 18 years or more: a randomized, open-label, active-controlled phase 3 trial.

Continuous emergence of new variants of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), enhanced transmissibility, significant immune escape, and waning immunity call for booster vaccination. We evaluated the safety, immunogenicity, and efficacy of heterologous booster with a SARS-CoV-2 mRNA vaccine SYS6006 versus an active control vaccine in a randomized, open-label, active-controlled phase 3 trial in healthy adults aged 18 years or more who had received two or three doses of SARS-CoV-2 inactivated vaccine in China. The trial started in December 2022 and lasted for 6 months. The participants were randomized (overall ratio: 3:1) to receive one dose of SYS6006 (N = 2999) or an ancestral receptor binding region-based, alum-adjuvanted recombinant protein SARS-CoV-2 vaccine (N = 1000), including 520 participants in an immunogenicity subgroup. SYS6006 boosting showed good safety profiles with most AEs being grade 1 or 2, and induced robust wild-type and Omicron BA.5 neutralizing antibody response on Days 14 and 28, demonstrating immunogenicity superiority versus the control vaccine and meeting the primary objective. The relative vaccine efficacy against COVID-19 of any severity was 51.6% (95% CI, 35.5-63.7) for any variant, 66.8% (48.6-78.5) for BA.5, and 37.7% (2.4-60.3) for XBB, from Day 7 through Month 6. In the vaccinated and infected hybrid immune participants, the relative vaccine efficacy was 68.4% (31.1-85.5) against COVID-19 of any severity caused by a second infection. All COVID-19 cases were mild. SYS6006 heterologous boosting demonstrated good safety, superior immunogenicity and high efficacy against BA.5-associated COVID-19, and protected against XBB-associated COVID-19, particularly in the hybrid immune population.Trial registration: Chinese Clinical Trial Registry: ChiCTR2200066941.

The role of Interleukin-22 in severe acute pancreatitis.

Severe acute pancreatitis (SAP) begins with premature activation of enzymes, promoted by the immune system, triggering a potential systemic inflammatory response that leads to organ failure with increased mortality and a bleak prognosis. Interleukin-22 (IL-22) is a cytokine that may have a significant role in SAP. IL-22, a member of the IL-10 cytokine family, has garnered growing interest owing to its potential tissue-protective properties. Recently, emerging research has revealed its specific effects on pancreatic diseases, particularly SAP. This paper provides a review of the latest knowledge on the role of IL-22 and its viability as a therapeutic target in SAP.

Finite element analysis of a new preoperative traction for cervical kyphosis: suspensory traction.

A finite element model of cervical kyphosis was established to analyze the stress of cervical spine under suspensory traction and to explore the mechanism and effect of it. A patient with typical cervical kyphosis (C2-C5) underwent CT scan imaging, and 3D slicer was used to reconstruct the C2 to T2 vertebral bodies. The reconstructed data was imported into Hypermesh 2020 and Abaqus 2017 for meshing and finite element analysis. The changes of the kyphotic angle and the von Mises stress on the annulus fibrosus of each intervertebral disc and ligaments were analyzed under suspensory traction conditions. With the increase of suspensory traction weight, the overall kyphosis of cervical spine showed a decreasing trend. The correction of kyphosis was mainly contributed by the change of kyphotic segments. The kyphotic angle of C2-C5 was corrected from 45° to 13° finally. In cervical intervertebral discs, the stress was concentrated to anterior and posterior part, except for C4-5. The stress of the anterior longitudinal ligament (ALL) decreased from the rostral to the caudal, and the high level von Mises stress of the kyphotic segments appeared at C2-C3, C3-C4, and C4-C5. The roles of the other ligaments were not obvious. The kyphotic angle was significantly reduced by the suspensory traction. Shear effect due to the high von Mises stress in the anterior and posterior parts of annulus fibrosus and the tension on the anterior longitudinal ligament play a role in the correction of cervical kyphosis.

Hedgehog ligand and receptor cooperatively regulate EGFR stability and activity in non-small cell lung cancer.

PURPOSE: The hyperactivation of epidermal growth factor receptor (EGFR) plays a crucial role in non-small cell lung cancer (NSCLC). Hedgehog (Hh) signaling has been implicated in the tumorigenesis and progression of various cancers, however, its function in NSCLC cells remains controversial. Herein, we present a novel finding that challenges the current understanding of Hh signaling in tumor growth. METHODS: Expression of Hh ligands and receptor were assessed using TCGA datasets, immunoblotting and immunohistochemical. Biological function of Hh ligands and receptor in NSCLC were tested using colony formation, cell count kit-8 (CCK-8) and xenograft assays. Biochemical effect of Hh ligands and receptor on regulating EGFR stability and activity were checked via immunoblotting. RESULTS: Expression of Hh ligands and receptor was suppressed in NSCLC tissues, and the lower expression levels of these genes were associated with poor prognosis. Ptch1 binds to EGFR and facilitates its poly-ubiquitylation and degradation independent of downstream transcriptional signaling. Moreover, Hh ligands cooperate with Ptch1 to regulate the protein stability and activity of EGFR. This unique mechanism leads to a suppressive effect on NSCLC tumor growth. CONCLUSION: Non-canonical Hh signaling pathway, involving cooperation between Hh ligands and their receptor Ptch1, facilitates the degradation of EGFR and attenuates its activity in NSCLC. These findings provide novel insights into the regulation of EGFR protein stability and activity, offer new diagnostic indicators for molecular typing of NSCLC and identify potential targets for targeted therapy of this challenging disease.

SARS-CoV-2 infection in pregnant patients on TNFα inhibitor: Real-life data with a review of literature.

Tumor necrosis factor alpha (TNFα) is involved in the occurrence of negative pregnancy outcomes. The study aimed to evaluate the safety and efficacy of the immunosuppressive TNFα inhibitors (TNFαi) in the treatment of patients with a history of recurrent reproductive failure in the context of COVID-19 pandemics. We reviewed 85 patients who received TNFαi (certolizumab pegol) during Mainland China's first wave of COVID-19 pandemic, from 21st Nov 2022-11 th Jan 2023. We also collected corresponding data from 130 pregnant patients who never used TNFαi for comparison. There were no significant differences in the history of previous pregnancy loss, miscarriage, embryo implantation failure, comorbidities and doses of COVID-19 vaccination. 82.2% and 87.7% pregnant patients contracted primary COVID-19 with symptoms in TNFαi group and no-TNFαi group. Duration of symptoms was significantly longer in TNFαi group and the incidences of cough and lethargy was significantly higher in TNFαi group. Both groups reported similar severity to same-aged close contacts, similar rates of other symptoms and hospitalization. No deaths were reported. In the in vitro fertilization (IVF) subgroup, we achieved a biochemical pregnancy loss rate of 17.4%, miscarriage rate of 21.7%, ongoing pregnancy rate and live birth rate of 34.2%. COVID-19 did not influence the live birth rate. We concluded that TNFαi administration in pregnancy was not associated with increased susceptivity to and severity of COVID-19. However, TNFαi users showed more prominent symptoms and longer recovery time. The pregnancy outcomes with TNFαi in such high-risk group for pregnancy loss was satisfactory.

Classification and prognostic factors of patients with cervical spondylotic myelopathy after surgical treatment: a cluster analysis.

Identifying potential prognostic factors of CSM patients could improve doctors' clinical decision-making ability. The study retrospectively collected the baseline data of population characteristics, clinical symptoms, physical examination, neurological function and quality of life scores of patients with CSM based on the clinical big data research platform. The modified Japanese Orthopedic Association (mJOA) score and SF-36 score from the short-term follow-up data were entered into the cluster analysis to characterize postoperative residual symptoms and quality of life. Four clusters were yielded representing different patterns of residual symptoms and quality of patients' life. Patients in cluster 2 (mJOA RR 55.8%) and cluster 4 (mJOA RR 55.8%) were substantially improved and had better quality of life. The influencing factors for the better prognosis of patients in cluster 2 were young age (50.1 ± 11.8), low incidence of disabling claudication (5.0%) and pathological signs (63.0%), and good preoperative SF36-physiological function score (73.1 ± 24.0) and mJOA socre (13.7 ± 2.8); and in cluster 4 the main influencing factor was low incidence of neck and shoulder pain (11.7%). We preliminarily verified the reliability of the clustering results with the long-term follow-up data and identified the preoperative features that were helpful to predict the prognosis of the patients. This study provided reference and research basis for further study with a larger sample data, extracting more patient features, selecting more follow-up nodes, and improving clustering algorithm.

Ultrasound-assisted fermentation of Porphyra yezoensis sauce at different growth stages using Lactiplantibacillus plantarum: Metabolic response and biological activity.

This study first employed ultrasonic-assisted fermentation of seaweed foot material with Lactiplantibacillus plantarum to produce Porphyra yezoensis sauce. The aim was to examine L. plantarum's growth and metabolism of nutritional components at different growth stages under low- (133.99 W/L) and high-ultrasonic power densities (169.17 W/L). After 24-h fermentation, L. plantarum exhibited a 21.32 % increase in the sonicated P. yezoensis sauce at 133.99 W/L and the logarithmic growth phase compared to that at 169.17 W/L. In addition, compared to the non-sonicated sauce, total phenolic and flavonoid contents increased by around 58 % and 27 % in sonicated sauce at 133.99 W/L, reaching 92.38 mg GEA/g DW and 111.08 mg RE/g DW, respectively. Principal Component Analysis (PCA) of the evaluation criteria for different fermentation stages under 133.99 W/L power ultrasonication revealed that the P. yezoensis sauce generated more phenolic compounds and exhibited stronger antioxidant capabilities in the sonicated sample at the logarithmic phase of L. plantarum. Compared to the traditional treated P. yezoensis sauce, the content of free amino acids was significantly increased in sonicated sauce, especially for logarithmic phase. Finally, GC-IMS analysis demonstrated that the ultrasonication at logarithmic phase released more volatile compounds compared to the non-sonicated sauce. This led to a reduction in the fishy odour of the Porphyra yezoensis sauce and an improved release of favourable flavour compounds.