FOXA3 regulates cholesterol metabolism to compensate for low uptake during the progression of lung adenocarcinoma.

Cholesterol metabolism is vital for multiple cancer progression, while how cholesterol affects lung, a low-cholesterol tissue, for cancer metastasis and the underlying mechanism remain unclear. In this study, we found that metastatic lung adenocarcinoma cells acquire cellular dehydrocholesterol and cholesterol by endogenous cholesterol biosynthesis, instead of uptake upon cholesterol treatment. Besides, we demonstrated that exogenous cholesterol functions as signaling molecule to induce FOXA3, a key transcription factor for lipid metabolism via GLI2. Subsequently, ChIP-seq analysis and molecular studies revealed that FOXA3 transcriptionally activated Hmgcs1, an essential enzyme of cholesterol biosynthesis, to induce endogenous dehydrocholesterol and cholesterol level for membrane composition change and cell migration. Conversely, FOXA3 knockdown or knockout blocked cholesterol biosynthesis and lung adenocarcinoma metastasis in mice. In addition, the potent FOXA3 inhibitor magnolol suppressed metastatic gene programs in lung adenocarcinoma patient-derived organoids (PDOs). Altogether, our findings shed light onto unique cholesterol metabolism and FOXA3 contribution to lung adenocarcinoma metastasis.

Impact of Lymph Node Dissection on Survival and Tumor Recurrence for Patients with Resected cT1-2N0 Small Cell Lung Cancer.

BACKGROUND: The role of lymph node (LN) dissection for early-stage small cell lung cancer (SCLC) remains unclear. Therefore, this study aimed to investigate the impact of LN dissection on patient survival and tumor recurrence for patients with cT1-2N0M0 SCLC. METHODS: Patients with cT1-2N0M0 SCLC who underwent intent-to-cure surgery at our institution between 2011 and 2019 were retrospectively reviewed. The survival outcomes of patients with systematic LN dissection (n = 112) and LN sampling (n = 35) were compared before and after propensity score-matching (PSM), as were the outcomes for patients with (pN+) and without (pN0) pathologic nodal metastasis. The prognostic impact of LN dissection was evaluated through multivariable analysis. RESULTS: The dissection group displayed significantly better overall survival (66.4% vs. 48.4%; P = 0.009) and recurrence-free survival (RFS) (63.5% vs. 37.6%;, P = 0.003) than the sampling group at 5 years. The rate of local recurrence was significantly lower in the dissection group than in the sampling group (11.6% vs. 42.9%; P < 0.001). After adjustment by multivariable analysis and PSM, LN dissection retained its independent favorable effects. The overall nodal upstaging rate was 37.6%, and the dissection group had a tendency of better RFS in both the pN0 (P = 0.05) and pN+ (P = 0.036) patients. CONCLUSION: Systematic LN dissection is associated with improved survival of patients who undergo surgery for cT1-2N0 SCLC. Further studies are warranted to verify the necessity of LN dissection in the surgery for SCLC.

[Research on "Augmented Reality" Assisted Precise Thoracoscopic Resection of Pulmonary Nodules].

To break with traditional preoperative localization, a porcine animal model to evaluate the safety of augmented reality (AR) assisted localization of solitary pulmonary nodules (SPN) was used. Before the experiment, Microsoft HoloLens AR system was used to bring the CT image into the laboratory after 3D reconstruction. The virtual model was fitted with real body surface markers, and the virtual positioning auxiliary line and auxiliary locator were used to perform puncture positioning before surgery. Data related to actual puncture path and expected planned path were recorded in the experiment. SPSS 26.0 was used to calculate the puncture accuracy under AR assisted positioning, and the results obtained were acceptable in segmentectomy or wedge pneumonectomy. Its feasibility in animal models will also be evaluated, and its safety and efficacy will need to be further studied in clinical trials.

A three-dimensional finite difference model for ocean acoustic propagation and benchmarking for topographic effects.

A three-dimensional (3D) finite difference (FD) model with formal fourth-order accuracy has been developed for the ocean acoustic Helmholtz equation (HE), which can be used to address arbitrary bathymetry and provide more accurate benchmark solutions for other 3D underwater acoustic approximate models. The derivatives in the acoustic HE are numerically discretized based on regular grids, and the perfectly matched layer is introduced to absorb unphysical reflections from the boundaries where Sommerfeld radiation conditions are deployed. The system of linear equations is solved using a parallel matrix-free geometric multigrid preconditioned biconjugate gradient stabilized iteration method, and the code (named COACH) is run on the Tianhe-2 supercomputer in China. Four 3D topographic benchmark acoustic cases-a wedge waveguide, Gaussian canyon, conical seamount, and corrugated seabed-are simulated to test the present FD model, and the maximum number of grid points reaches 33.15 × 109 in the wedge waveguide case, running in parallel with 988 central processing unit cores. Furthermore, the accuracy and generality of the present model have been verified by solution comparisons with other available 3D acoustic propagation models, and the two-dimensional and 3D transmission loss contours are presented to facilitate the distinguishing among the acoustic field features of these cases.

Two new secolignans from the roots of Urtica fissa.

Two new secolignans, 3,4-trans-3-hydroxymethyl-4-[bis(4-hydroxy-3- methoxyphenyl)methyl]butyrolactone (1) and 3,4-trans-3-hydroxymethyl-4- [bis(3,4-dimethoxyphenyl)methyl]butyrolactone (2) have been isolated from the roots of Urtica fissa E.Pritz. Their structures were determined on the basis of spectroscopic methods, especially 1H NMR, 13C NMR, 2D NMR, and HR-ESI-MS. The inhibitory effects on N1 and N2, two subtypes of neuraminidases (NAs), of these two compounds were assayed.

Extended Right Thoracic Approach Compared With Limited Left Thoracic Approach for Patients With Middle and Lower Esophageal Squamous Cell Carcinoma: Three-year Survival of a Prospective, Randomized, Open-label Trial.

OBJECTIVE: To investigate whether survival is improved by using the right thoracic approach (extended lymphadenectomy) compared with the left thoracic approach (limited lymphadenectomy) for esophageal cancer. BACKGROUND: The optimal surgical technique for esophageal cancer remains unclear. METHODS: Between May 2010 and July 2012, 300 patients with middle and lower thoracic esophageal carcinoma were randomized to receive esophagectomy through either the right or left thoracic approach. Of these, 286 patients with squamous cell carcinoma determined by postoperative pathology were included in this analysis. Disease-free survival (DFS) and overall survival (OS) were compared between the right (n = 146) and left thoracic groups (n = 140). RESULTS: The median follow-up was 55.9 months [95% confidence interval (CI): 53.1-58.6]. The 3-year DFS rates were 62% and 52% in the right and left thoracic arms, respectively [hazard ratio (HR) 0.709; 95% CI, 0.506-0.995; P = 0.047, log-rank test]. The 3-year OS rates were 74% and 60%, respectively (HR, 0.663; 95% CI, 0.457-0.961; P = 0.029). Subgroup analyses revealed longer DFS in the right thoracic arm (vs left thoracic arm) in patients with lymph node involvement (HR, 0.632; 95% CI, 0.412-0.969, P = 0.034), but not in patients without lymph node involvement (HR, 0.757; 95% CI, 0.434-1.320, P = 0.325), and in patients with R1-2 resection margins (HR, 0.495; 95% CI, 0.290-0.848, P = 0.009), but not R0 margins (HR, 0.944; 95% CI, 0.603-1.477, P = 0.801). CONCLUSIONS: Compared with the left thoracic approach, the right thoracic approach associated with increased DFS and OS in esophageal squamous cell carcinoma patients, particularly in those with lymph node involvement and/or R1-2 resection margins.

Prevalence and Clinicopathological Characteristics of BRAF Mutations in Chinese Patients with Lung Adenocarcinoma.

BACKGROUND: This study was designed to identify the prevalence of BRAF mutations in Chinese patients with lung adenocarcinoma, and to reveal the association between BRAF mutations and clinicopathological characteristics in these patients. METHODS: From October 2007 to February 2013, patients with newly diagnosed primary lung adenocarcinoma were detected for mutations in BRAF, EGFR, KRAS, HER2 and ALK. Clinicopathological characteristics, including sex, age, TNM stage, tumor differentiation, smoking status, histological subtypes, and survival information were analyzed. RESULTS: Of 1358 patients with lung adenocarcinoma, 20 patients were harboring BRAF mutations, including five BRAF V600E mutations and 15 BRAF non-V600E mutations. Among these, BRAF N581I and BRAF G593S were newly reported. BRAF mutations were associated with smoking status (odds ratio 3.28; 95 % CI 1.33-8.08; p = 0.008). In patients less than 60 years of age, BRAF mutations tended to have poor differentiation in tumor samples (70.0 vs. 35.1 %; p = 0.014), and were more likely to relapse (70 vs. 28 %; p = 0.008). A significant difference was found in relapse-free survival (RFS) between BRAF mutations and other mutations, but not in overall survival. CONCLUSIONS: The prevalence of BRAF mutations in Chinese patients with lung adenocarcinoma was approximately 1.5 %. BRAF mutations were more frequent in current smokers. Patients harboring BRAF mutations had a higher rate of recurrence and worse RFS compared with other patients.

Thermal, water, and land cover factors led to contrasting urban and rural vegetation resilience to extreme hot months.

With continuing global warming and urbanization, it is increasingly important to understand the resilience of urban vegetation to extreme high temperatures, but few studies have examined urban vegetation at large scale or both concurrent and delayed responses. In this study, we performed an urban-rural comparison using the Enhanced Vegetation Index and months that exceed the historical 90th percentile in mean temperature (referred to as "hot months") across 85 major cities in the contiguous United States. We found that hot months initially enhanced vegetation greenness but could cause a decline afterwards, especially for persistent (≥4 months) and intense (≥+2 °C) episodes in summer. The urban responses were more positive than rural in the western United States or in winter, but more negative during spring-autumn in the eastern United States. The east-west difference can be attributed to the higher optimal growth temperatures and lower water stress levels of the western urban vegetation than the rural. The urban responses also had smaller magnitudes than the rural responses, especially in deciduous forest biomes, and least in evergreen forest biomes. Within each biome, analysis at 1 km pixel level showed that impervious fraction and vegetation cover, local urban heat island intensity, and water stress were the key drivers of urban-rural differences. These findings advance our understanding of how prolonged exposure to warm extremes, particularly within urban environments, affects vegetation greenness and vitality. Urban planners and ecosystem managers should prioritize the long and intense events and the key drivers in fostering urban vegetation resilience to heat waves.

Patient-derived tumor-like cell clusters for personalized chemo- and immunotherapies in non-small cell lung cancer.

Many patient-derived tumor models have emerged recently. However, their potential to guide personalized drug selection remains unclear. Here, we report patient-derived tumor-like cell clusters (PTCs) for non-small cell lung cancer (NSCLC), capable of conducting 100-5,000 drug tests within 10 days. We have established 283 PTC models with an 81% success rate. PTCs contain primary tumor epithelium self-assembled with endogenous stromal and immune cells and show a high degree of similarity to the original tumors in phenotypic and genotypic features. Utilizing standardized culture and drug-response assessment protocols, PTC drug-testing assays reveal 89% overall consistency in prospectively predicting clinical outcomes, with 98.1% accuracy distinguishing complete/partial response from progressive disease. Notably, PTCs enable accurate prediction of clinical outcomes for patients undergoing anti-PD1 therapy by combining cell viability and IFN-γ value assessments. These findings suggest that PTCs could serve as a valuable preclinical model for personalized medicine and basic research in NSCLC.

The learning curve on uniportal video-assisted thoracoscopic lobectomy with the help of postoperative review of videos.

Objectives: Video-assisted thoracoscopic lobectomy has become the preferred surgical approach in experienced centers, and uniportal approaches are becoming increasingly used. But the uniportal approach is still not widely applied presumably due to the learning difficulties of this complex procedure. The use of surgical videos may be helpful to accelerate the learning of this new techniques as in other fields. In this study, we aimed to analyze the learning curve of uniportal video-assisted thoracoscopic lobectomy with the help of postoperative review of videos. Methods: 114 patients with early-stage lung cancer who underwent uniportal video-assisted thoracoscopic lobectomy performed from 2020 to 2021 were reviewed in this study. We recorded the operation video for each patient and reviewed all the videos after surgery. The learning curves were assessed using cumulative sum analysis and the collected data of perioperative outcomes were assessed. Results: The CUMSUM curve showed its inflection points were around case 38 and 53. It was less compared with previous studies, which about 57-140 cases are needed to attain the proficient phase. The perioperative outcomes were similar in each phase, which included intraoperative blood loss (79.00 ± 26.70 vs 70.67 ± 26.64 vs 70.56 ± 27.23, p=0.0119), the length of hospital stay (3.60 ± 1.52 days vs. 3.23 ± 0.90 days vs. 3.06 ± 0.88 days, p=0.053), the rate of prolonged air leak and conversion to open thoracotomy. There was also no significant difference in the numbers and station of lymph node dissection among the three phases. Conclusions: Uniportal video-assisted thoracoscopic lobectomy is a safe and reliable approach. Recording and reviewing the operation video could help the surgeon to improve deficiencies and refine the procedure.

The application of patient-derived organoid in the research of lung cancer.

Lung cancer is the most common cancer and the leading cause of cancer-related death worldwide. However, mechanisms of its progression remained unclear and new treatments against this disease are rapidly emerging. As a novel preclinical model, patient-derived organoid (PDO) can also be established from the patient's tumor tissue and cultured in the laboratory, which preserves the key biological characteristics of the original tumor. Compared to the patient-derived xenograft (PDX) model of lung cancer, the culture success rate is improved, and the time and cost of model establishment are largely reduced. PDO is also expected to provide a more individual model to predict the efficacy of anti-cancer treatment in vitro. This paper summarizes the current application of PDO in the translational research of lung cancer.

Bioinformatic analysis of single-cell RNA sequencing dataset dissects cellular heterogeneity of triple-negative breast cancer in transcriptional profile, splicing event and crosstalk network.

BACKGROUND: Triple-negative breast cancer (TNBC) is a subtype of breast cancer with high tumoral heterogeneity, while the detailed regulatory network is not well known. METHODS: Via single-cell RNA-sequencing (scRNA-seq) data analysis, we comprehensively investigated the transcriptional profile of different subtypes of TNBC epithelial cells with gene regulatory network (GRN) and alternative splicing (AS) event analysis, as well as the crosstalk between epithelial and non-epithelial cells. RESULTS: Of note, we found that luminal progenitor subtype exhibited the most complex GRN and splicing events. Besides, hnRNPs negatively regulates AS events in luminal progenitor subtype. In addition, we explored the cellular crosstalk among endothelial cells, stromal cells and immune cells in TNBC and discovered that NOTCH4 was a key receptor and prognostic marker in endothelial cells, which provide potential biomarker and target for TNBC intervention. CONCLUSIONS: In summary, our study elaborates on the cellular heterogeneity of TNBC, revealing that NOTCH4 in endothelial cells was critical for TNBC intervention. This in-depth understanding of epithelial cell and non-epithelial cell network would provide theoretical basis for the development of new drugs targeting this sophisticated network in TNBC.

Perioperative outcomes of bi-pigtail catheter drainage strategy versus conventional chest tube after uniportal video-assisted thoracic lung surgery.

OBJECTIVES: Chest tube (CT) drainage is a main cause of postoperative pain in lung surgery. Here, we introduced a novel drainage strategy with bi-pigtail catheters (PCs) and conducted a randomized controlled trial to compare with conventional CT drainage after uniportal video-assisted thoracic surgery lung surgery. METHODS: A single-centre, prospective, open-labelled, randomized controlled trial (ChiCTR2000035337) was conducted with a preplanned sample size of 396. The primary outcome was the numerical pain rating scale (NPRS) on the first postoperative day. Secondary outcomes included other indicators of postoperative pain, drainage volume, duration of drainage, postoperative hospital stay, incidence of postoperative complications, CT reinsertion and medical costs. RESULTS: A total number of 396 patients were randomized between August 2020 and January 2021, 387 of whom were included in the final analysis. The baseline and clinical characteristics of the patients were well balanced between 2 groups. The NPRS on the first postoperative day was significantly lower in the PC group than in the CT group (2.40 ± 1.27 vs 3.02 ± 1.39, p < 0.001), as well as the second/third-day NPRS, the incidence of sudden severe pain (9/192, 4.7% vs 34/195, 17.4%, P < 0.001) and pain requiring intervention (19/192, 9.9% vs 46/195, 23.6%, P < 0.001). In addition, the medical cost in the PC group was lower (US$7809 ± 1646 vs US$8205 ± 1815, P = 0.025). Univariable and multivariable analyses revealed that the drainage strategy was the only factor influencing the incidence of pain requiring intervention. CONCLUSIONS: The drainage strategy with bi-PCs in patients undergoing uniportal video-assisted thoracic surgery lung surgery alleviates postoperative pain with adequate safety and efficacy.

Multiple factors affecting perioperative antibiotherapy in minimally invasive lung surgery: a retrospective case-control study.

Background: During the perioperative period, empiric antibiotic escalation therapy is not uncommon, leading to longer postoperative hospital stay and higher medical expenses. However, few systematic studies have investigated this issue. In this case-control study, we explored the factors affecting empiric antibiotic escalation therapy after minimally invasive lung surgery during hospitalization, with the aim to enhance recovery after surgery and rational use of antibiotics. Methods: The data of patients who underwent minimally invasive lung surgery at our center from January 2019 to December 2020 were collected from the hospital information system (HIS). A total of 1,360 cases were identified based on the inclusion and exclusion criteria. These patients were divided into 2 groups: group A (n=825), in which patients did not receive antibiotic escalation therapy after surgery; and group B (n=535), in which patients received empiric antibiotic escalation therapy after surgery. A logistic regression model was used to analyze the influencing factors of empiric antibiotherapy escalation during postoperative hospitalization. Results: Compared with group A, group B had significantly longer postoperative hospital stay (5.05±2.78 vs. 4.49±2.24 days, P<0.001) and slightly higher average total hospitalization costs (74,080.85±23,796.51 vs. 71,798.09±21,307.26 yuan, P=0.067). Multivariate analysis showed that the statistically significant factors included history of secondary lung surgery [odds ratio (OR): 3.267; 95% confidence interval (CI): 1.305-8.178; P=0.011], preoperative hemoglobin A1c (HbA1c) ≥6.5% (OR: 1.603; 95% CI: 1.143-2.249; P=0.006), postoperative fever of unknown origin [temperature (T) >38 ℃; OR: 2.494; 95% CI: 1.321-4.708; P=0.005], postoperative hypoalbuminemia (intravenous albumin administration for ≥2 days; OR: 14.125; 95% CI: 1.777-112.282; P=0.012). Conclusions: Multivariate analysis showed that history of secondary lung surgery, preoperative HbA1c ≥6.5%, postoperative fever of unknown origin (T >38 ℃), and postoperative hypoalbuminemia (<35 g/L and intravenous albumin administration ≥2 days) were the independent risk factors for empiric antibiotic escalation therapy after minimally invasive lung surgery. For preoperative risk factors, further cohort studies should be conducted to explore better intervention indicators or measures. For postoperative risk factors, perioperative dynamic monitoring of procalcitonin can guide the rational use of antibiotics, reduce the risk of drug resistance and hospitalization cost.

Expert consensus on multi-disciplinary treatment, whole-course pulmonary rehabilitation management in patients with lung cancer and chronic obstructive lung disease.

Comorbidity of lung cancer and chronic obstructive pulmonary disease (COPD) is very common. Surgical operation is the initial treatment of lung cancer. But surgery operation will aggravate the symptoms of COPD, such as shortness of breath, chest tightness. On the other side, the COPD also increase the perioperative complications. Besides, the COPD may also influence the anti-cancer treatment and long-term survival of lung cancer patients. At present, there are guidelines for pulmonary rehabilitation (PR) of COPD or lung cancer respectively, but there is no reference expert consensus on the PR of patients with lung cancer who are comorbidity of COPD. Primary care has to satisfy the patient's complex needs holistically, and single-disease guidelines are unsuitable. In view of this, we organized experts from respiratory department, thoracic surgery department, oncology department, nursing department, etc., to write the expert consensus. We discussed the contents of the expert consensus through literature review, expert correspondence, expert meeting and discussion. This expert consensus contain five parts: introduction, respiratory assessment, timing of PR, PR strategies, perioperative PR management strategies in lung cancer patients with COPD. This expert consensus focuses on patients with COPD comorbid lung cancer and undergoing surgery operation, highlighting the concept of whole process management. For clinical medical staff, this expert consensus will promote the practice of PR in and out the hospital for this specific patient; for patients, this expert consensus is helpful to better understand PR and improve the enthusiasm of participating in PR in the whole process.

Homologous recombination deficiency (HRD) can predict the therapeutic outcomes of immuno-neoadjuvant therapy in NSCLC patients.

BACKGROUND: Neoadjuvant immunotherapy is emerging as novel effective intervention in lung cancer, but study to unearth effective surrogates indicating its therapeutic outcomes is limited. We investigated the genetic changes between non-small cell lung cancer (NSCLC) patients with varied response to neoadjuvant immunotherapy and discovered highly potential biomarkers with indicative capability in predicting outcomes. METHODS: In this study, 3 adenocarcinoma and 11 squamous cell carcinoma NSCLC patients were treated by neoadjuvant immunotherapy with variated regimens followed by surgical resection. Treatment-naive FFPE or fresh tissues and blood samples were subjected to whole-exome sequencing (WES). Genetic alternations were compared between differently-responded patients. Findings were further validated in multiple public cohorts. RESULTS: DNA damage repair (DDR)-related InDel signatures and DDR-related gene mutations were enriched in better-responded patients, i.e., major pathological response (MPR) group. Besides, MPR patients exhibited provoked genome instability and unique homologous recombination deficiency (HRD) events. By further inspecting alternation status of homology-dependent recombination (HR) pathway genes, the clonal alternations were exclusively enriched in MPR group. Additionally, associations between HR gene alternations, percentage of viable tumor cells and HRD event were identified, which orchestrated tumor mutational burden (TMB), mutational intratumor heterogeneity (ITH), somatic copy number alteration (SCNA) ITH and clonal neoantigen load in patients. Validations in public cohorts further supported the generality of our findings. CONCLUSIONS: We reported for the first time the association between HRD event and enhanced neoadjuvant immunotherapy response in lung cancer. The power of HRD event in patient therapeutic stratification persisted in multifaceted public cohorts. We propose that HR pathway gene status could serve as novel and additional indicators guiding immune-neoadjuvant and immunotherapy treatment decisions for NSCLC patients.

A uniportal right upper lobectomy by three-arm robotic-assisted thoracoscopic surgery using the da Vinci (Xi) Surgical System in the treatment of early-stage lung cancer.

Minimally invasive techniques, typified by video-assisted thoracoscopic surgery, are widely practiced in the treatment of thoracic diseases all around the world, and video-assisted thoracoscopic surgery has been recognized as a standard treatment method for early staged lung cancer. Among them, robotic-assisted thoracoscopic surgery, which has the advantages of providing a three-dimensional view and better maneuverability, has emerged as a next-generation technique in the field of minimally invasive surgery and is also gaining its popularity with the idea of Enhanced Recovery After Surgery deeply rooted in patients' minds. Up to now, robotic-assisted thoracoscopic surgery usually requires 3 or 4 ports with 1 or 2 additional access incisions. Meanwhile, traditional video-assisted thoracoscopic surgery can now be completed with uniportal method, with less postoperative pain and higher patient satisfaction with respect to the number of incisions in comparison with the multi-port technique. To inform the integration of these new minimally invasive techniques, here, we present a case in which uniportal right upper lobectomy was performed using the 4th generation da Vinci Robotic Surgical System (Xi). With continuous innovation in robotic minimally invasive techniques and improvements in surgical skills, we believe more patients will benefit from robotic-assisted thoracoscopic surgery with single port in the near future.

Augmented reality-assisted localization of solitary pulmonary nodules for precise sublobar lung resection: a preliminary study using an animal model.

BACKGROUND: Accurate localization of early lung cancer, manifested as solitary pulmonary nodules (SPNs) on computed tomography (CT), is critical in sublobar lung resection. The AR-assisted localization of SPNs was evaluated using a pig animal model. METHODS: A Microsoft HoloLens AR system was used. First, a plastic thoracic model was used for the pilot study. Three female 12 months 45 kg Danish Landrace Pigs were then used for the animal study. Thirty natural pulmonary structures, such as lymphonodus and bifurcated bronchioles or bronchial vessels, were chosen as simulated SPNs. The average angle between the actual puncturing needle and the expected path, the average distance between the puncture point and the plan point, and the difference between the actual puncturing depth and expected depth were recorded, and the accuracy rate was calculated. RESULTS: The point selected in the plastic thoracic model could be hit accurately with the assistance from the AR system in the pilot study. Moreover, the average angle between the actual puncturing needle and the expected path was 14.52°±6.04°. Meanwhile, the average distance between the puncture point and the expected point was 8.74±5.07 mm, and the difference between the actual and expected depths was 9.42±7.95 mm. Puncturing within a 1 cm3 area around the SPN using a hook-wire was considered a successful hit. The puncture accuracy was calculated. The average hit rate within a spherical area with a diameter of 1 cm range was 76.67%, and within a diameter of 2 cm range was 100%. CONCLUSIONS: The HoloLens AR-assisted localization of SPNs may become a promising technique to improve the surgical treatment of early-stage lung cancer. Here, we evaluated its feasibility in an animal model. Nevertheless, its safety and effectiveness require further investigation in clinical trials.