Non-invasive diagnosis of pulmonary nodules by circulating tumor DNA methylation: A prospective multicenter study.

BACKGROUND: With the popularization of computed tomography, more and more pulmonary nodules (PNs) are being detected. Risk stratification of PNs is essential for detecting early-stage lung cancer while minimizing the overdiagnosis of benign nodules. This study aimed to develop a circulating tumor DNA (ctDNA) methylation-based, non-invasive model for the risk stratification of PNs. METHODS: A blood-based assay ("LUNG-TRAC") was designed to include novel lung cancer ctDNA methylation markers identified from in-house reduced representative bisulfite sequencing data and known markers from the literature. A stratification model was trained based on 183 ctDNA samples derived from patients with benign or malignant PNs and validated in 62 patients. LUNG-TRAC was further single-blindly tested in a single- and multi-center cohort. RESULTS: The LUNG-TRAC model achieved an area under the curve (AUC) of 0.810 (sensitivity = 74.4 % and specificity = 73.7 %) in the validation set. Two test sets were used to evaluate the performance of LUNG-TRAC, with an AUC of 0.815 in the single-center test (N = 61; sensitivity = 67.5 % and specificity = 76.2 %) and 0.761 in the multi-center test (N = 95; sensitivity = 50.7 % and specificity = 80.8 %). The clinical utility of LUNG-TRAC was further assessed by comparing it to two established risk stratification models: the Mayo Clinic and Veteran Administration models. It outperformed both in the validation and the single-center test sets. CONCLUSION: The LUNG-TRAC model demonstrated accuracy and consistency in stratifying PNs for the risk of malignancy, suggesting its utility as a non-invasive diagnostic aid for early-stage peripheral lung cancer. CLINICAL TRIAL REGISTRATION: www. CLINICALTRIALS: gov (NCT03989219).

Shape-sensing Robotic-assisted Bronchoscopy (SS-RAB) in Sampling Peripheral Pulmonary Nodules: A Prospective, Multicenter Clinical Feasibility Study in China.

BACKGROUND: The ION system is a shape-sensing robotic-assisted bronchoscopy (SS-RAB) platform developed to biopsy peripheral pulmonary nodules (PPNs). There is a lack of data describing the use of this system in the Chinese population. The study aimed to assess the feasibility and safety of using SS-RAB to diagnose PPNs across multiple centers within China. METHODS: This prospective, multicenter study used SS-RAB in consecutive patients with solid or sub-solid PPNs 8 to 30 mm in largest diameter. Primary endpoints were diagnostic yield and the rates of procedure- or device-related complications. Radial endobronchial ultrasound (rEBUS) was to confirm lesion localization, followed by sampling, using the Flexision biopsy needle, biopsy forceps, and cytology brush. Subjects with nonmalignant index biopsy results were followed up to 6 months. RESULTS: A total of 90 PPNs were biopsied from 90 subjects across 3 centers using SS-RAB. The median nodule size was 19.4 mm (IQR: 19.3, 24.6) in the largest dimension. In all (100%) cases, the catheter successfully reached the target nodule with tissue samples obtained. The diagnostic yield was 87.8% with a sensitivity for malignancy of 87.7% (71/81). In a univariate analysis, nodule lobar location, presence of bronchus sign, and rEBUS view were associated with a diagnostic sample, but only rEBUS view showed an association in a multivariate analysis. The overall pneumothorax rate was 1.1% without pneumothorax requiring intervention, and there was no periprocedural bleeding. CONCLUSION: As an emerging technology in the Chinese population, SS-RAB can safely biopsy PPNs with strong diagnostic performance.

KCNE4 is a crucial host factor for Orf virus infection by mediating viral entry.

The orf virus (ORFV) poses a serious threat to the health of domestic small ruminants (i.e., sheep and goats) and humans on a global scale, causing around $150 million in annual losses to livestock industry. However, the host factors involved in ORFV infection and replication are still elusive. In this study, we compared the RNA-seq profiles of ORFV-infected or non-infected sheep testicular interstitial cells (STICs) and identified a novel host gene, potassium voltage-gated channel subfamily E member 4 (KCNE4), as a key host factor involved in the ORFV infection. Both RNA-seq data and RT-qPCR assay revealed a significant increase in the expression of KCNE4 in the infected STICs from 9 to 48 h post infection (hpi). On the other hand, the RT-qPCR assay detected a decrease in ORFV copy number in both the STICs transfected by KCNE4 siRNA and the KCNE4 knockout (KO) HeLa cells after the ORFV infection, together with a reduced fluorescence ratio of ORFV-GFP in the KO HeLa cells at 24 hpi, indicating KCNE4 to be critical for the ORFV infection. Furthermore, the attachment and internalization assays showed decreased ORFV attachment, internalization, replication, and release by the KO HeLa cells, demonstrating a potential inhibition of ORFV entry into the cells by KCNE4. Pretreatment with the KCNE4 inhibitors such as quinidine and fluoxetine significantly repressed the ORFV infection. All our findings reveal KCNE4 as a novel host regulator of the ORFV entry and replication, shedding new insight into the interactive mechanism of ORFV infection. The study also highlights the K+ channels as possible druggable targets to impede viral infection and disease.

A risk-scoring model based on endobronchial ultrasound multimodal imaging for predicting metastatic lymph nodes in lung cancer patients.

Background and Objectives: Endobronchial ultrasound (EBUS) imaging is a valuable tool for predicting lymph node (LN) metastasis in lung cancer patients. This study aimed to develop a risk-scoring model based on EBUS multimodal imaging (grayscale, Doppler mode, elastography) to predict LN metastasis in lung cancer patients. Patients and Methods: This retrospective study analyzed 350 metastatic LNs in 314 patients with lung cancer and 124 reactive LNs in 96 patients with nonspecific inflammation. The sonographic findings were compared with the final pathology results and clinical follow-up. Univariate and multivariate logistic regression analyses were performed to evaluate the independent risk factors of metastatic LNs. According to the ß coefficients of corresponding indicators in logistic regression analysis, a risk-scoring model was established. Receiver operating characteristic curve was applied to evaluate the predictive capability of model. Results: Multivariate analysis showed that short axis >10 mm, distinct margin, absence of central hilar structure, presence of necrosis, nonhilar vascularity, and elastography score 4 to 5 were independent predictors of metastatic LNs. Both short axis and margin were scored 1 point, and the rest of independent predictors were scored 2 points. The combination of 3 EBUS modes had the highest area under the receiver operating characteristic and accuracy of 0.884 (95% confidence interval, 0.846-0.922) and 87.55%, respectively. The risk stratification was as follows: 0 to 2 points, malignancy rate of 11.11%, low suspicion; 3 to 10 points, malignancy rate of 86.77%, high suspicion. Conclusions: The risk-scoring model based on EBUS multimodal imaging can effectively evaluate metastatic LNs in lung cancer patients to support clinical decision making.

Prognostic predictive value of Ki-67 in stage I-II triple-negative breast cancer.

Aim: Our research aimed to determine an optimal cutoff value and investigate the prognostic predictive function of Ki-67. Materials & methods: We retrospectively enrolled 1146 patients diagnosed with stage I-II triple-negative breast cancer. Disease-free and overall survival were analyzed using the Kaplan-Meier method and the Cox regression model. Results: We classified Ki-67 >45% as the high group (n = 716). A Ki-67 level of >45% was associated with poorer disease-free survival (p = 0.039) and overall survival (p = 0.029). Lymph node stage, neoadjuvant chemotherapy, and radiotherapy were independent predictive variables of prognosis. Conclusion: Triple-negative breast cancer may be further subcategorized according to the Ki-67 level. Neoadjuvant chemotherapy and postoperative radiotherapy can improve the prognosis of early triple-negative breast cancer.

This study aimed to find the best value of Ki-67, which is a marker used in breast cancer. At last, according to the Ki-67 level over 45%, triple-negative breast cancer may be divided into two groups. Based on the level of Ki-67, treatment decisions may be better. However, we still need more studies to confirm this.

Triple-negative breast cancer may be further subcategorized according to the Ki-67 level >45%, which is associated with a poorer prognosis. Treatment decisions based on the level of Ki-67 may be more favorable to the prognosis of patients.

nCas9 Engineering for Improved Target Interaction Presents an Effective Strategy to Enhance Base Editing.

Base editors (BEs) are a recent generation of genome editing tools that couple a cytidine or adenosine deaminase activity to a catalytically impaired Cas9 moiety (nCas9) to enable specific base conversions at the targeted genomic loci. Given their strong application potential, BEs are under active developments toward greater levels of efficiency and safety. Here, a previously overlooked nCas9-centric strategy is explored for enhancement of BE. Based on a cytosine BE (CBE), 20 point mutations associated with nCas9-target interaction are tested. Subsequently, from the initial positive X-to-arginine hits, combinatorial modifications are applied to establish further enhanced CBE variants (1.1-1.3). Parallel nCas9 modifications in other versions of CBEs including A3A-Y130F-BE4max, YEE-BE4max, CGBE, and split-AncBE4max, as well as in the context of two adenine BEs (ABE), likewise enhance their respective activities. The same strategy also substantially improves the efficiencies of high-fidelity nCas9/BEs. Further evidence confirms that the stabilization of nCas9-substrate interactions underlies the enhanced BE activities. In support of their translational potential, the engineered CBE and ABE variants respectively enable 82% and 25% higher rates of editing than the controls in primary human T-cells. This study thus demonstrates a highly adaptable strategy for enhancing BE, and for optimizing other forms of Cas9-derived tools.

Correlation Between Coronal Position Sequence of Lumbar and WOMAC Score in Knee Osteoarthritis (KOA) in Standard Standing Position.

INTRODUCTION: Sagittal sequences of the spine have been shown to correlate with knee osteoarthritis (KOA), but coronal sequences and KOA have never been studied before. The study required patients to use a standard standing posture and aimed to explore the relationship between coronal position of lumbar spine and WOMAC score in KOA. METHODS: This is a cross-sectional observational study. Data on a total of 268 patients with KOA were collected. Patients were photographed in a standard standing position and lumbar-sacrum offset distance (L-SOD) and lumbar-knee offset distance (ΔL-KOD) were calculated. Patients were then divided into different groups according to different critical values and differences were compared. RESULTS: In the L-SOD of L1-3, WOMAC function (P = 0.021, P = 0.032, P = 0.001) and total score (P = 0.039, P = 0.034, P < 0.001) were different. In the L-SOD of L3-4, WOMAC pain score were different (P = 0.001, P = 0.032). At a cutoff of 13 mm, ΔL-KOD of L1-2 showed significant differences in pain part (P = 0.025, P = 0.039) and total score (P = 0.036, P = 0.050). There were significant differences in pain (P = 0.023, P = 0.027, P = 0.022), function (P = 0.048, P = 0.038, P = 0.047), and total score (P = 0.030, P = 0.027, P = 0.029) of L3-5. In the 18-mm cutoff group, only L1 and L2 have differences in the pain part (P = 0.050, P = 0.038). CONCLUSION: Coronal balance of the lumbar spine is associated with knee pain and function. The pelvis plays an important role in maintaining the coronal balance. Both the lumbar spine and the knee joint should be considered when developing the surgical strategy.

As a result of population aging, the number of patients suffering from both knee osteoarthritis (KOA) and degenerative diseases of the lumbar spine is increasing. It has been reported that patients with KOA have less symptomatic recovery after lumbar surgery, and that patients with lumbar degenerative disease have less symptomatic improvement after knee surgery than those without lumbar disease. So the knee and lumbar must be interacting in some way. Previous studies have confirmed the correlation between lumbar sagittal position sequence and KOA. However, no studies have been conducted on coronal sequences and KOA of the lumbar spine. We believe that it is because patients are required to stand naturally when taking coronal x-rays, and natural standing will lead to individual differences in the distance between the feet of patients, thereby preventing analysis. In our study, for the first time, we used a uniform stance to avoid this effect. The advantage of uniform stance is that individual differences can be excluded, and the same patient can be compared before and after treatment (because the natural stance of the patient's feet will be different before and after treatment), which is greatly conducive to the study. Our research found that the offset of the lumbar spine in the coronal position and the distance between the central vertical line of the lumbar spine and the bilateral knee joint are significantly correlated with knee pain and function. This may have some guiding significance for lumbar and knee surgery. For lumbar surgery (such as degenerative scoliosis), previous studies have suggested that short segment fixation is sufficient for patients with small Cobb angle. However, according to our conclusion, this may cause accelerated knee joint degeneration in the patient's later stages, which requires the surgeon to comprehensively evaluate the condition of the patient's knee and lumbar spine, and then formulate surgical strategies. The same is true for knee surgery: previous studies have shown no significant correlation between knee deformity and pain. Therefore, for patients with knee deformity and accompanying pain, knee surgery may not be the best choice, and it is more important to correct the deviation of the spine.

Potential of Thermal Ablation Combined with Immunotherapy in Peripheral Lung Tumors: A Review and Prospect.

BACKGROUND: Lung tumors are prevalent malignancies associated with a high mortality rate, imposing significant medical and societal burdens. Although immunotherapy shows promise in improving survival, response rates are relatively modest. Thermal ablation can not only eliminate tumor cells directly but also enhance antitumor immunity response, thus manifesting a remarkable propensity to synergize with immunotherapy. SUMMARY: In this review, we provided a brief overview of the application of thermal ablation in peripheral lung tumors. We summarized the patient selection of thermal ablation. We highlighted the potential of thermal ablation to augment the antitumor immune response, offering a promising avenue for combined therapies. We summarized studies assessing the synergistic effects of thermal ablation and immunotherapy in preclinical and clinical settings. Lastly, we underscored the urgent issues that warrant in-depth exploration when applying thermal ablation and immunotherapy to lung tumor patients. KEY MESSAGES: This review emphasized the prospects of using thermal ablation combined with immunotherapy in patients with peripheral lung tumors. However, further research is needed to enhance and optimize this treatment strategy.

Chinese expert consensus on cone-beam CT-guided diagnosis, localization and treatment for pulmonary nodules.

Cone-beam computed tomography (CBCT) system can provide real-time 3D images and fluoroscopy images of the region of interest during the operation. Some systems can even offer augmented fluoroscopy and puncture guidance. The use of CBCT for interventional pulmonary procedures has grown significantly in recent years, and numerous clinical studies have confirmed the technology's efficacy and safety in the diagnosis, localization, and treatment of pulmonary nodules. In order to optimize and standardize the technical specifications of CBCT and guide its application in clinical practice, the consensus statement has been organized and written in a collaborative effort by the Professional Committee on Interventional Pulmonology of China Association for Promotion of Health Science and Technology.

Robotic-assisted bronchoscopy for the diagnosis of peripheral pulmonary lesions: A systematic review and meta-analysis.

Robotic-assisted bronchoscopy (RAB) is a newly developed bronchoscopic technique for the diagnosis of peripheral pulmonary lesions (PPLs). The objective of this meta-analysis was to analyze the diagnostic yield and safety of RAB in patients with PPLs. Five databases (PubMed, Embase, Web of Science, CENTRAL, and ClinicalTrials.gov) were searched from inception to April 2023. Two independent investigators screened retrieved articles, extracted data, and assessed the study quality. The pooled diagnostic yield and complication rate were estimated. Subgroup analysis was used to explore potential sources of heterogeneity. Publication bias was assessed using funnel plots and the Egger test. Sensitivity analysis was also conducted to assess the robustness of the synthesized results. A total of 725 lesions from 10 studies were included in this meta-analysis. No publication bias was found. Overall, RAB had a pooled diagnostic yield of 80.4% (95% CI: 75.7%-85.1%). Lesion size of >30 mm, presence of a bronchus sign, and a concentric radial endobronchial ultrasound view were associated with a statistically significantly higher diagnostic yield. Heterogeneity exploration showed that studies using cryoprobes reported better yields than those without cryoprobes (90.0%, 95% CI: 83.2%-94.7% vs. 79.0%, 95% CI: 75.8%-82.2%, p < 0.01). The pooled complication rate was 3.0% (95% CI: 1.6%-4.4%). In conclusion, RAB is an effective and safe technique for PPLs diagnosis. Further high-quality prospective studies still need to be conducted.

Emerging Light-Harvesting Materials Based on Organic Photovoltaic D/A Heterojunctions for Efficient Photocatalytic Water Splitting.

Photocatalytic water splitting to hydrogen is a highly promising method to meet the surging energy consumption globally through the environmentally friendly means. As the initial step before photocatalysis, harvesting photons from sunlight is crucially important, thus making the design of photosensitizers with visible even near-infrared (NIR) absorptions get more and more attentions. In the past three years, organic donor/acceptor (D/A) heterojunctions with absorptions extending to 950 nm, have emerged as the new star light-harvesting materials for photocatalytic water splitting, demonstrating exciting advantages over inorganic materials in solar light utilization, hydrogen yielding rate, etc. This Minireview firstly gives a brief discussion about the principle processes and determining factors for photocatalytic water splitting with organic photovoltaic D/A heterojunction as photosensitizers. Thereafter, the current progress is summarized in details by introducing typical and excellent D/A heterojunction-based photocatalytic systems. Finally, not only the great prospects but also the most challenging issues confronted by organic D/A heterojunctions are indicated along with a perspective on the opportunities and new directions for future material explorations.

Cryoablation triggers type I interferon-dependent antitumor immunity and potentiates immunotherapy efficacy in lung cancer.

BACKGROUND: Cryoablation is a minimally invasive option for patients with medically inoperable non-small cell lung cancer (NSCLC) and can trigger abscopal immune-regulatory effects. However, it remains unclear how cryoablation affects the host-level immune response in NSCLC. In this study, we investigated the local and systemic immunological effects of cryoablation and the potential of combining cryoablation with programmed cell death protein 1 (PD-1) blockade to boost immunotherapy efficacy in NSCLC. METHODS: We first investigated systemic immunological effects induced by cryoablation in patients with early-stage NSCLC. Subsequently, we explored cryoablation-induced antitumor immunity and the underlying biological mechanisms using KP (Kras G12D/+, Tp53 -/-) mutant lung cancer cell allograft mouse models. Moreover, the synergistic efficacy of cryoablation and PD-1 blockade was explored in both mouse models and patients with unresectable NSCLC. RESULTS: We found that cryoablation significantly increased circulating CD8+ T cell subpopulations and proinflammatory cytokines in patients with early-stage NSCLC. In lung cancer cell allograft mouse models, we demonstrated that cryoablation resulted in abscopal growth inhibition of contralateral, non-ablated tumors. Integrated analysis of bulk, single-cell RNA and T cell receptor (TCR) sequencing data revealed that cryoablation reprogrammed the intratumoral immune microenvironment and increased CD8+ T cell infiltration with higher effector signature, interferon (IFN) response, and cytolytic activity. Mechanistically, cryoablation promoted antitumor effect through the STING-dependent type I IFN signaling pathway, and type I IFN signaling blockade attenuated this antitumor effect. We also found that the combination of PD-1 blockade with cryoablation further inhibited tumor growth compared with either treatment alone in an allograft mouse model. Moreover, the combination therapy induced notable tumor suppression and CD8+ T cell infiltration in patients with unresectable NSCLC. CONCLUSIONS: Our results provide mechanistic insights into how cryoablation triggers the antitumor immune effect in lung cancer, thereby potentiating programmed cell death ligand 1 (PD-L1)/PD-1 blockade efficacy in the clinical treatment of NSCLC.

Transbronchial cryoablation in peripheral lung parenchyma with a novel thin cryoprobe and initial clinical testing.

BACKGROUND: Transbronchial cryoablation shows potential as a local therapy for inoperable peripheral lung cancer. However, its clinical application for peripheral pulmonary lesions has not been reported yet. METHODS: An improved cryoprobe with an 8-mm-long, 1.9-mm-wide cryotip was used. Initially, the safety and effectiveness of this cryoprobe were assessed in an in vivo porcine model. Transbronchial cryoablation with 2 or 3 freeze-thaw cycles (10 min or 15 min in each freezing time) was performed in 18 pigs under CT monitoring. Radiological and pathological examinations were performed to evaluate the extent of cryoablation. Subsequently, nine patients with stage IA peripheral lung cancer or metastases underwent transbronchial cryoablation with this cryoprobe under the guidance of navigation bronchoscopy and cone-beam CT. Technical success, safety and outcomes were assessed. RESULTS: 36 cryoablation procedures were performed successfully without any major complications in the porcine model. The extent of cryoablation increased with freezing time and the number of freeze-thaw cycles, which peaked at 24 hours and then gradually decreased. Pathological results showed a change from massive haemorrhage at 24 hours to fibrous hyperplasia with chronic inflammation after 4 weeks. In the clinical trial, 10 cryoablations were performed on 9 tumours with a technical success rate of 100%. One mild treatment-related complication occurred. Of the nine tumours, seven achieved complete ablation, while two exhibited incomplete ablation and subsequent local progression at 6 months. CONCLUSION: Our initial experience indicated that transbronchial cryoablation was a safe and feasible procedure for non-surgical peripheral stage IA lung cancer or pulmonary metastases. TRIAL REGISTRATION NUMBER: ChiCTR2200061544.

Robotic-assisted bronchoscopy in the diagnosis of peripheral pulmonary lesions.

More peripheral pulmonary lesions (PPLs) are detected by low-dose helical computed tomography (CT) either incidentally or via dedicated lung cancer screening programs. Thus, using methods for safe and accurate diagnosis of these lesions has become increasingly important. Transthoracic needle aspiration (TTNA) and transbronchial lung biopsy (TBLB) are routinely performed during the diagnostic workup for PPLs. However, TTNA often carries the risk of pneumothorax, uncontrollable airway hemorrhage, and does not allow mediastinal staging in one procedure. In contrast, traditional TBLB often has a poorer diagnostic yield despite fewer complications. With the ongoing development of technology applied to bronchoscopy, guided bronchoscopy has become widely used and the diagnostic yield of TBLB has improved. Additionally, guided bronchoscopy continues to demonstrate a better safety profile than TTNA. In recent years, robotic-assisted bronchoscopy (RAB) has been introduced and implemented in the diagnosis of PPLs. At present, RAB has two platforms that are commercially available: Monarch™ and Ion™; several other platforms are under development. Both systems differ in characteristics, advantages, and limitations and offer features not seen in previous guided bronchoscopy. Several studies, including cadaveric model studies and clinical trials, have been conducted to examine the feasibility and performance of RAB using these two systems; large multicenter studies are underway. In this review, published experimental results, focusing on diagnostic yield and complications of RAB, are analyzed and the potential clinical application of RAB is discussed, which will enable the operators to have a clear overview of RAB.