Allicin affects immunoreactivity of osteosarcoma cells through lncRNA CBR3-AS1.

Objective: To analyze the effect of allicin on the immunoreactivity of osteosarcoma (OS) cells and further explore whether its mechanism is related to the long non-coding Ribonucleic Acid (lncRNA) CBR3-AS1/miR-145-5p/GRP78 axis, so as to provide clinical evidence. Methods: The human OS cell line Saos-2 was treated with allicin at 25, 50, and 100 µmol/L, respectively, to observe changes in cell biological behaviors. Subsequently, CBR3-AS1 abnormal expression vectors were constructed and transfected into Saos-2 to discuss their influence on OS. Furthermore, the regulatory relationship between allicin and the CBR3-AS1/miR-145-5p/GRP78 axis was validated by rescue experiments. Finally, a nude mice tumorigenesis experiment was carried out to analyze the effects of allicin and CBR3-AS1/miR-145-5p/GRP78 axis on the growth of living tumors. Alterations in T-lymphocyte subsets were also detected to assess the effect of allicin on OS immunoreactivity. Results: With the increase of allicin concentration, Saos-2 activity decreased and apoptosis increased (P < 0.05). In addition, the expression of CBR3-AS1 and GRP78 decreased after allicin intervention, while miR-145-5p increased (P < 0.05). Silencing CBR3-AS1 led to reduced Saos-2 activity, enhanced apoptosis, and activated mitophagy and endoplasmic reticulum stress (P < 0.05). In the rescue experiment, the effect of CBR3-AS1 on OS cells was reversed by silencing miR-145-5p, while the impact of miR-145-5p was reversed by GRP78. Finally, the tumorigenesis experiment in nude mice confirmed the regulatory effects of allicin and CBR3-AS1/miR-145-5p/GRP78 on tumor growth in vivo. Meanwhile, it was seen that allicin activated CD4+CD8+ in OS mice, confirming that allicin has the effect of activating OS immunoreactivity. Conclusions: Allicin activates OS immunoreactivity and induces apoptosis through the CBR3-AS1/miR-145-5p/GRP78 molecular axis.

Pathological and imaging features of pulmonary invasive mucinous adenocarcinoma-a retrospective cohort study.

Background: Pulmonary invasive mucinous adenocarcinoma (IMA) is a rare subtype of lung cancer which is easily misdiagnosed as inflammatory nodules, tuberculosis, pulmonary diffuse lesions, or hamartomas due to the lack of clinical specificity. This study aims to identify the pathological and imaging characteristics of IMA, which will favor to improve the diagnostic and therapeutic efficacy. Methods: A retrospective study was conducted by enrolling patients histopathologically diagnosed with pulmonary IMA in the current study between January 2014 and December 2021. The clinical pathological and radiological data were collected for analysis to evaluate the radiological patterns and pathological and molecular characteristics of IMA. Results: A total of 136 patients were included in the study, of whom 58 were male and 78 were female. The patients had an average age of 63.0±9.7 years. The tumors were classified into the following three pathological types: pure mucinous (76 cases) featured by only mucinous cells observed under the microscope; mixed mucinous (23 cases) featured as an attached-wall, papillary, acinar, and solid tumor cells with more than 10% mucinous cells.; and mucinous-absent (29 cases) featured with the absence of mucous cells, but still can detect more than 10% of mucin expresses. In terms of the morphological classification based on the CT scans, 88 (64.7%) cases were identified as the nodular type, 31 (22.8%) as the inflammatory type, 15 (11.1%) as the mass-like type, and two (1.5%) as the diffuse type. For the molecular features, patients afflicted with IMA showed much lower levels of thyroid transcription factor-1 (15%) than those with usual adenocarcinoma (over 80%). However, cytokeratin 20 was more common in IMA (50%) than the usual adenocarcinoma (about 5%). The K-RAS mutation was prevalent in 75% of IMA, which contrasted sharply to its occurrence in a mere 15% of the usual adenocarcinoma. Epidermal growth factor receptor mutations were rarer in IMA (less than 5%) than the usual adenocarcinoma (about 50%). Conclusions: The pathological and imaging features enrich our understanding of the disease's heterogeneity, which will contribute to more personalized diagnostic and therapeutic strategies.

Evaluation of the preoperative neutrophil-to-lymphocyte ratio as a predictor of the micropapillary component of stage IA lung adenocarcinoma.

OBJECTIVE: To assess the ability of markers of inflammation to identify the solid or micropapillary components of stage IA lung adenocarcinoma and their effects on prognosis. METHODS: We performed a retrospective study of clinicopathologic data from 654 patients with stage IA lung adenocarcinoma collected between 2013 and 2019. Logistic regression analysis was used to identify independent predictors of these components, and we also evaluated the relationship between markers of inflammation and recurrence. RESULTS: Micropapillary-positive participants had high preoperative neutrophil-to-lymphocyte ratios. There were no significant differences in the levels of markers of systemic inflammation between the participants with or without a solid component. Multivariate analysis showed that preoperative neutrophil-to-lymphocyte ratio (odds ratio [OR] = 2.094; 95% confidence interval [CI], 1.668-2.628), tumor size (OR = 1.386; 95% CI, 1.044-1.842), and carcinoembryonic antigen concentration (OR = 1.067; 95% CI, 1.017-1.119) were independent predictors of a micropapillary component. There were no significant correlations between markers of systemic inflammation and the recurrence of stage IA lung adenocarcinoma. CONCLUSIONS: Preoperative neutrophil-to-lymphocyte ratio independently predicts a micropapillary component of stage IA lung adenocarcinoma. Therefore, the potential use of preoperative neutrophil-to-lymphocyte ratio in the optimization of surgical strategies for the treatment of stage IA lung adenocarcinoma should be further studied.

[Cangxi Tongbi Capsules promote chondrocyte autophagy by regulating circRNA＿0008365/miR-1271/p38 MAPK pathway to inhibit development of knee osteoarthritis].

To investigate the mechanism by which Cangxi Tongbi Capsules promote chondrocyte autophagy to inhibit knee osteoarthritis(KOA) progression by regulating the circRNA＿0008365/miR-1271/p38 mitogen-activated protein kinase(MAPK) pathway. The cell and animal models of KOA were established and intervened with Cangxi Tongbi Capsules, si-circRNA＿0008365, si-NC, and Cangxi Tongbi Capsules combined with si-circRNA＿0008365. Flow cytometry and transmission electron microscopy were employed to determine the level of apoptosis and observe autophagosomes, respectively. Western blot was employed to reveal the changes in the protein levels of microtubule-associated protein light chain 3(LC3)Ⅱ/Ⅰ, Beclin-1, selective autophagy junction protein p62/sequestosome 1, collagen Ⅱ, a disintegrin and metalloproteinase with thrombospondin motifs 5(ADAMTS-5), and p38 MAPK. The mRNA levels of circRNA＿0008365, miR-1271, collagen Ⅱ, and ADAMTS-5 were determined by qRT-PCR. Hematoxylin-eosin staining was employed to reveal the pathological changes of the cartilage tissue of the knee, and enzyme-linked immunosorbent assay to measure the levels of interleukin-1ß(IL-1ß) and tumor necrosis factor-alpha(TNF-α). The chondrocytes treated with IL-1ß showed down-regulated expression of circRNA＿0008365, up-regulated expression of miR-1271 and p38 MAPK, lowered autophagy level, increased apoptosis rate, and accelerated catabolism of extracellular matrix. The intervention with Cangxi Tongbi Capsules up-regulated the expression of circRNA＿0008365, down-regulated the expression of miR-1271 and p38 MAPK, increased the autophagy level, decreased the apoptosis rate, and weakened the catabolism of extracellular matrix. However, the effect of Cangxi Tongbi Capsules was suppressed after interfering with circRNA＿0008365. The in vivo experiments showed that Cangxi Tongbi Capsules dose-dependently inhibited the p38 MAPK pathway, enhanced chondrocyte autophagy, and mitigated articular cartilage damage and inflammatory response, thereby inhibiting the progression of KOA in rats. This study indicated that Cangxi Tongbi Capsules promoted chondrocyte autophagy by regulating the circRNA＿0008365/miR-1271/p38 MAPK pathway to inhibit the development of KOA.

[Growth Regularity of Pulmonary Ground Glass Nodules Based on 3D Reconstruction Technology].

BACKGROUND: Since the popularization of computed tomography (CT) technology, the detection rate of pulmonary ground glass nodules (GGNs) with imaging follow-up as the main management method has increased significantly. The purpose of this study is to quantitatively analyze the changes of pulmonary GGNs during the follow-up process with three-dimensional reconstruction technology, explore the natural progression of pulmonary GGNs, and provide effective basis for clinical guidance for patients to conduct reasonable management of nodules. METHODS: A total of 115 cases of pulmonary GGNs with regular follow-up in the Combined Outpatient Department of Zhoushan Hospital from March 2015 to November 2022 were enrolled. Quantitative imaging features of nodules were extracted by semi-automatic segmentation of 3D Slicer software to evaluate the growth of nodules and clinical intervention during follow-up. RESULTS: The average baseline age of the patients was (56.9±10.1) yr. The mean follow-up time was (48.8±18.9) months. The two-dimensional diameter of baseline CT scan was (7.9±2.9) mm, and the maximum three-dimensional diameter was (10.1±3.4) mm. The two-dimensional diameter of the last CT scan was (9.9±4.7) mm, and the maximum three-dimensional diameter was (11.4±5.1) mm. A total of 27 cases (23.5%) showed an increase during follow-up, with a median volume doubling time of 822 days and a median mass doubling time of 1,007 days. 32 cases were surgically resected, including 6 cases of invasive adenocarcinoma (IAC), 16 cases of minimally invasive adenocarcinoma (MIA), 8 cases of adenocarcinoma in situ (AIS) and 2 cases of atypical adenomatous hyperplasia (AAH). Five nodules underwent surgical intervention due to the progression of two-dimensional diameter, which was pathologically confirmed as pre-invasive lesions, but their three-dimensional maximum diameter showed no significant change. Nodular morphology, lobulated sign, spiculated sign and vacuole signs all promoted the growth of nodules in univariate analysis. There were significant differences in age, baseline diameter, mean CT value, median CT value, 10% and 90% percentile CT number between the growth group and the stable group (P<0.05). Multivariate Logistic regression analysis showed that age and average CT value were risk factors for nodule growth (P<0.05). Receiver-operating characteristic (ROC) curve analysis results indicated that the age ≥63 years old, the baseline three-dimensional maximum diameter ≥9.2 mm, and the average CT value ≥-507.8 HU were more likely to accelerate the growth of GGNs. The maximum three-dimensional diameter ≥14.4 mm and the average CT value ≥-495.7 HU may be a higher malignant probability. CONCLUSIONS: GGNs show an inert growth process, and the use of three-dimensional measurements during follow-up is of greater significance. For persistent glass grinding nodules ≥63 years old, the baseline three-dimensional maximum diameter ≥9.2 mm, and the average CT value ≥-507.8 HU are more likely to increase. However, most nodules still have good prognosis after progression, and long-term follow-up is safe.

Treatment of bisphenol pollutant in water by N,P-co-doped carbon nanosheet: Fast degradation, toxicity elimination and reaction mechanism investigation.

Metal-free carbon catalysts perform well in peroxymonosulfate-based advanced oxidation process for the treatment of organic pollutant-containing wastewater. Herein, a natural biomolecule of adenosine triphosphate (ATP), containing abundant N and P elements, served as sole precursor to prepare N,P-co-doped carbon through one-step anoxic pyrolysis, which was applied as peroxymonosulfate activator to treat bisphenol-contaminated water. Owing to the endogenous N and P elements in ATP, in-situ doping was achieved for the prepared carbon material with excellent doping effect, such as high doping amount and numerous defects. During pyrolysis process, the generated gases facilitated the exfoliation of carbon structure, resulting in a nanosheet-like morphology with large specific surface area, e.g., 852.75 m2 g-1 for NPCN-900 sample obtained at 900 °C. Benefiting from the structural modulation brought by N,P co-doping, typical sample of NPCN-900 presented excellent catalytic performance towards bisphenol AF (BPAF) degradation through PMS activation. An apparent reaction rate constant of 0.4115 min-1 was calculated under the investigated reaction conditions. Further studies indicated that 1O2, surface-bound •OH and SO4-• worked together in NPCN-900/PMS system for BPAF degradation. Graphitic N, pyrrolic N, CO groups, defect structure and the doped P atoms in NPCN-900 contributed to PMS activation. It was also important that the toxicity of BPAF solution could be preliminarily eliminated after treatment by NPCN-900/PMS system, which was verified by ecotoxicity assessments through ECOSAR program and green algae growth experiments.

LC-MS/MS Based Volatile Organic Compound Biomarkers Analysis for Early Detection of Lung Cancer.

(1) Background: lung cancer is the world's deadliest cancer, but early diagnosis helps to improve the cure rate and thus reduce the mortality rate. Annual low-dose computed tomography (LD-CT) screening is an efficient lung cancer-screening program for a high-risk population. However, LD-CT has often been characterized by a higher degree of false-positive results. To meet these challenges, a volatolomic approach, in particular, the breath volatile organic compounds (VOCs) fingerprint analysis, has recently received increased attention for its application in early lung cancer screening thanks to its convenience, non-invasiveness, and being well tolerated by patients. (2) Methods: a LC-MS/MS-based volatolomics analysis was carried out according to P/N 5046800 standard based breath analysis of VOC as novel cancer biomarkers for distinguishing early-stage lung cancer from the healthy control group. The discriminatory accuracy of identified VOCs was assessed using subject work characterization and a random forest risk prediction model. (3) Results: the proposed technique has good performance compared with existing approaches, the differences between the exhaled VOCs of the early lung cancer patients before operation, three to seven days after the operation, as well as four to six weeks after operation under fasting and 1 h after the meal were compared with the healthy controls. The results showed that only 1 h after a meal, the concentration of seven VOCs, including 3-hydroxy-2-butanone (TG-4), glycolaldehyde (TG-7), 2-pentanone (TG-8), acrolein (TG-11), nonaldehyde (TG-19), decanal (TG-20), and crotonaldehyde (TG-22), differ significantly between lung cancer patients and control, with the invasive adenocarcinoma of the lung (IAC) having the most significant difference. (4) Conclusions: this novel, non-invasive approach can improve the detection rate of early lung cancer, and LC-MS/MS-based breath analysis could be a promising method for clinical application.

miR-31/QKI-5 axis facilitates cell cycle progression of non-small-cell lung cancer cells by interacting and regulating p21 and CDK4/6 expressions.

BACKGROUND: RNA-binding protein Quaking-5 (QKI-5), a major isoform of QKIs, inhibits tumor progression in non-small cell lung cancer (NSCLC). However, the underlying molecular mechanisms of QKI-5 in the cell cycle of NSCLC are still largely unknown. METHODS: MTT, flow cytometry, and colony formation assays were used to investigate cellular phenotypic changes. Mice xenograft model was used to evaluate the antitumor activities of QKI-5. Co-immunoprecipitation, RNA immunoprecipitation (RIP), and RIP sequencing were used to investigate protein-protein interaction and protein-mRNA interaction. RESULTS: The QKI-5 expression was downregulated in NSCLC tissues compared with that in paired normal adjacent lung tissues. Overexpression of QKI-5 inhibited NSCLC cell proliferative and colony forming ability. In addition, QKI-5 induced cell cycle arrest at G0/G1 phase through upregulating p21Waf1/Cip1 (p21) expression and downregulating cyclin D1, cyclin-dependent kinase 4 (CDK4), and CDK6 expressions. Further analyses showed that QKI-5 interacts with p21 protein and CDK4, CDK6 mRNAs, suggesting a critical function of QKI-5 in cell cycle regulation. In agreement with in vitro study, the mouse xenograft models validated tumor suppressive functions of QKI-5 in vivo through altering cell cycle G1-phase-associated proteins. Moreover, we demonstrated that QKI-5 is a direct target of miR-31. The QKI-5 expression was anticorrelated with the miR-31 expression in NSCLC patient samples. CONCLUSION: Our results suggest that the miR-31/QKI-5/p21-CDK4-CDK6 axis might have critical functions in the progression of NSCLC, and targeting this axis could serve as a potential therapeutic strategy for NSCLC.

Risk factors for prolonged preoperative waiting time of intertrochanteric fracture patients undergoing operative treatment.

PURPOSE: Intertrochanteric fracture is a common fracture in older adults. We observed the case characteristics of intertrochanteric fracture and analyzed the risk factors for prolonged preoperative waiting time based on patient data from a 6 year period. Investigate the post-admission treatment of intertrochanteric fracture. METHODS: We retrospectively reviewed the medical records from July 2015 to July 2021 of patients hospitalized for intertrochanteric fracture who had undergone internal fixation surgery in the orthopedic ward of the Affiliated Hospital of Shandong University of Traditional Chinese Medicine. Data regarding gender, age, AO/OTA classification, preoperative waiting time, preoperative medical comorbidities, and complicated deep venous thrombosis (DVT) of lower limbs were collected. Statistical tests were used to evaluate the factors influencing preoperative preparation time and DVT. RESULTS: A total of 1812 cases were retrospectively analyzed, 1258 patients (69.43%) had three or more medical comorbidities. The average preoperative waiting time was 5.09 ± 3.27 days. Advanced age, more preoperative medical comorbidities and DVT led to longer preoperative waiting times, and preoperative medical comorbidities were an independent risk factor. Patients with advanced age and preoperative medical comorbidities were more likely to have DVT. CONCLUSION: Age and preoperative medical comorbidities are risk factors for DVT and prolonged preoperative preparation time in intertrochanteric fracture patients. Preoperative medical comorbidities are an independent risk factors affecting the preoperative waiting time, and a combination of multiple comorbidities almost predicts the delay of the operation time.

Allicin Inhibits Osteosarcoma Growth by Promoting Oxidative Stress and Autophagy via the Inactivation of the lncRNA MALAT1-miR-376a-Wnt/ß-Catenin Signaling Pathway.

Allicin, an organic sulfur compound extracted from the bulb of Allium sativum, can potentially prevent various tumors. Our previous study found that allicin can effectively suppress the proliferation of osteosarcoma cells. However, the molecular mechanisms have not been illustrated. In this study, Saos-2 and U2OS osteosarcoma cells were used to investigate the underlying mechanisms. A series of experiments were carried out to authenticate the anticancer property of allicin. Knockdown of lncRNA MALAT1 inhibited the proliferation, invasion and migration and promoted apoptosis of osteosarcoma cells. Knockdown of miR-376a increased the proliferation, invasion, and migration and dropped apoptosis of osteosarcoma cells. Furthermore, knockdown of miR-376a reversed the influences of MALAT1 silencing in osteosarcoma cells. Based on our data, MALAT1 could downregulate the expression of miR-376a, subsequently accelerating osteosarcoma. Moreover, oxidative stress and autophagy were identified as the potential key pathway of allicin. Allicin inhibited osteosarcoma growth and promoted oxidative stress and autophagy via MALATI-miR-376a. We also found that allicin promotes oxidative stress and autophagy to inhibit osteosarcoma growth by inhibiting the Wnt/ß-catenin pathway in vivo and in vitro. All data showed that allicin promotes oxidative stress and autophagy of osteosarcoma via the MALATI-miR-376a-Wnt/ß-catenin pathway.

Impact of minimal solid and micropapillary components on invasive lung adenocarcinoma recurrence.

BACKGROUND: The specific impacts of solid and micropapillary components on prognosis in lung adenocarcinoma remain unclear. Herein, we elucidated their distinct contributions to lung adenocarcinoma recurrence. MATERIALS AND METHODS: Lung adenocarcinoma was classified into solid and micropapillary absent (S-M-); solid absent, micropapillary present (S-M+); micropapillary absent, solid present (S + M-); and solid and micropapillary present (S + M+). Cumulative incidence of recurrence (CIR) was calculated using competing risk analysis. RESULTS: Of 994 adenocarcinomas, 650 (65.4%) were classified as S-M-; 152 (15.3%), S-M+; 148 (14.9%), S + M-; and 44 (4.4%), S + M+. In total, 168 (16.9%) patients had recurrence; 16 (1.6%) died from other causes. S-M- had significantly lower CIR than other groups (S-M- vs. S-M+: P < 0.001, S-M- vs. S + M-: P < 0.001, S-M- vs. S + M+: P < 0.001); S + M- had significantly higher CIR than S-M+ (P = 0.002). These differences remained significant in multivariable analysis. In stage IA, S-M- had significantly lower CIR than other groups (S-M- vs. S-M+: P = 0.006, S-M- vs. S + M-: P < 0.001, S-M- vs. S + M+: P < 0.001); S + M- and S + M+ had significantly higher CIR than S-M+ (P = 0.005, P = 0.008, respectively). These differences remained significant in multivariable analysis. CIR was not significantly different between S + M- and S-M+ subgroups. CONCLUSIONS: The presence of solid or micropapillary component (≥1%) was an independent risk factor for CIR; patients with solid component alone had a higher CIR than those with micropapillary component alone. In IA lung adenocarcinoma, patients with both solid and micropapillary components had a higher CIR than those with micropapillary component alone; the proportion of solid or micropapillary component was not associated with CIR.

Differentiation of adenocarcinoma in situ with alveolar collapse from minimally invasive adenocarcinoma or invasive adenocarcinoma appearing as part-solid ground-glass nodules (≤ 2 cm) using computed tomography.

PURPOSE: To investigate the differentiating computed tomographic (CT) features between adenocarcinoma in situ (AIS) with alveolar collapse and minimally invasive adenocarcinoma (MIA) or invasive adenocarcinoma (IA) appearing as part-solid nodules. METHODS: A total of 147 consecutive patients with 157 pathology-confirmed part-solid ground-glass nodules (GGNs) ≤ 20 mm without other pathological condition such as inflammation and fibrosis who underwent chest CT were included. RESULTS: The 157 part-solid GGNs included 33 (21.02%) pathologically confirmed AISs with alveolar collapse. Multivariate analysis revealed that smaller lesion size (odds ratio [OR] 0.671), and well-defined border (OR 5.544), concentrated distribution (OR 7.994), and homogeneity of the solid portion (OR 4.365) were significant independent predictors for differentiating AIS with alveolar collapse from MIA (P < 0.05) with excellent accuracy (area under receiver operating characteristic [ROC] curve, 0.902). Multivariate analysis revealed that smaller lesion size (OR 0.782), and size (OR 0.821), well-defined border (OR 5.752), and homogeneity of solid portion (OR 6.182) were significant independent predictors differentiating AIS with alveolar collapse from IA (P < 0.05) with excellent accuracy (area under ROC curve 0.910). CONCLUSION: Among part-solid GGNs, AIS with alveolar collapse can be accurately differentiated from MIA on the basis of smaller lesion size, well-defined border, concentrated distribution, and homogeneity of solid portion, and from IA according to smaller lesion size, and smaller size, well-defined border, and homogeneity of solid portion.

Lentinan as a natural stabilizer with bioactivities for preparation of drug-drug nanosuspensions.

Lentinan is a natural ß-glucan with various bioactivities and is combined with chemotherapy drugs for cancer treatment. Regorafenib is an oral multi-kinase inhibitor approved by FDA for treatment of metastatic colorectal cancer, advanced hepatocellular carcinoma, and metastatic gastrointestinal stromal tumors. Regorafenib has poor water solubility and multiple toxicities. We report drug-drug nanosuspensions of regorafenib and lentinan. Results of dynamic light scattering and scanning electron microscopy showed that the mean particle size of the regorafenib-lentinan nanosuspensions was approximately 200 nm and was uniformly distributed. Transmission electron microscopy findings indicated that lentinan stabilized the nanosuspensions by steric manner. Hydrogen bonds and hydrophobic interactions were found between regorafenib and lentinan by molecular dynamics simulation. The results of cytotoxicity assay and pharmacokinetics study in rats showed that the regorafenib-lentinan nanosuspensions reduced the toxicity and enhanced the in vitro anticancer activity and oral bioavailability of regorafenib. Lentinan as a natural stabilizer has the potential using for drug nanosuspensions. Drug-drug nanosuspensions are a new form of combination therapies that can reduce the number of drugs taken by patients and improve their compliance.

Intraoperative localization in minimally invasive surgery for small pulmonary nodules: a retrospective study.

BACKGROUND: Small pulmonary nodules are increasingly detected at an earlier stage and need to be removed via video-assisted thoracoscopic surgery (VATS). However, small pulmonary nodules are often difficult to locate during VATS and are typically nonvisible and nonpalpable on the lung surface. A variety of localization techniques have been developed. Here, we explored the application of an intraoperative body surface localization (IOBSL) and/or anatomical landmark localization (ALL) in minimally invasive surgery for small pulmonary nodules. METHODS: A total of 174 patients with small pulmonary nodules were divided into 3 groups: an IOBSL group, an ALL group, and an IOBSL+ALL group. VATS partial pneumonectomy was performed after the nodule localization, and the need for pulmonary segmentectomy/lobectomy and lymph node dissection was assessed according to the results of intraoperative rapid frozen section diagnosis. The duration, accuracy, and complications of each localization method were recorded and analyzed. RESULTS: ALL had shorter distance to the nodules (P=0.0282) but longer localization duration (P<0.05) than did IOBSL. The IOBSL+ALL group had higher localization accuracy than did the other 2 groups (P=0.0003) but with longer localization duration (P<0.001). No intraoperative complications were noted. CONCLUSIONS: The intraoperative technique has high localization accuracy and a low complication rate. It can be applied in VATS for pulmonary nodules, depending on the specific locations of the nodules.

Cancer-associated fibroblasts, matrix metalloproteinase-9 and lymphatic vessel density are associated with progression from adenocarcinoma in situ to invasive adenocarcinoma of the lung.

The present study aimed to investigate the roles of cancer-associated fibroblasts (CAFs), matrix metalloproteinase-9 (MMP-9) and lymphatic vessel density (LVD) during the progression from adenocarcinoma in situ (AIS) to invasive lung adenocarcinoma (IAC). A total of 77 patients with stage 0-IA lung adenocarcinoma were enrolled. The expression levels of α-smooth muscle actin, MMP-9 and D2-40 were immunohistochemically analyzed. Survival analysis was performed using the Kaplan-Meier method. In the non-invasive component, the proportion of CAFs and the expression levels of MMP-9 increased from AIS to IAC; however, the LVD was not significantly different. CAFs were positively correlated with levels of MMP-9. The LVD had no significant correlation with CAFs and MMP-9. In the invasive component, CAFs, MMP-9 and LVD were significantly higher in IAC compared with in minimally invasive adenocarcinoma. CAFs, MMP-9 and LVD were all positively correlated with each other. The micropapillary subtype in IAC was associated with overall survival (OS). The LVD in IAC, but not MMP-9 and CAFs, was associated with OS. CAFs, MMP-9 and LVD were involved in the progression from AIS to IAC. CAFs exhibited a strong association with MMP-9 levels in the non-invasive and invasive components. The increase in the proportion of CAFs and the expression levels of MMP-9 may have been an early event before the adenocarcinoma became invasive. Once the adenocarcinoma was invasive, the LVD served an important role in tumor invasion and metastasis, and hence may be used as a prognostic marker of poor OS in stage IA IAC.

Protein corona of metal-organic framework nanoparticals: Study on the adsorption behavior of protein and cell interaction.

Nanoscale metal-organic frameworks (NMOFs) have attracted considerable attention for controlled drug delivery. However, the interaction between nanoparticles and the biological macromolecules of physiological system must be valued because the formed protein corona will endow NMOFs with new biorecognition properties. In this study, we carried out detailed protein adsorption studies in vitro and cell uptake tests of HeLa cells for nanospherical Uio66 and nanooctahedral Uio67. Uio67 with higher binding constants to human serum albumin needed to combine more protein molecules to achieve colloidal stability state than that needed by Uio66, and this phenomenon led Uio67 to aggregate under the same incubation condition due to the formation of a single-layer protein. Uio67 also induced an evident conformation change in protein to stabilize the combination. In particular, the cell uptake efficiencies of the two systems showed a significant thickness dependence on the protein corona. When samples incubated in 10% fetal bovine serum (FBS), the intracellular rate was the highest for both systems, but the rate was not proportional to the FBS concentration. Results of this work are important to the development of the considerable potential NMOFs-based medicals and also provide additional insight into protein corona.

A Prediction Rule for Overall Survival in Non-Small-Cell Lung Cancer Patients with a Pathological Tumor Size Less Than 30 mm.

We sought to develop and validate a clinical nomogram model for predicting overall survival (OS) in non-small-cell lung cancer (NSCLC) patients with resected tumors that were 30 mm or smaller, using clinical data and molecular marker findings. We retrospectively analyzed 786 NSCLC patients with a pathological tumor size less than 30 mm who underwent surgery between 2007 and 2017 at our institution. We identified and integrated significant prognostic factors to build the nomogram model using the training set, which was subjected to the internal data validation. The prognostic performance was calibrated and evaluated by the concordance index (C-index) and risk group stratification. Multivariable analysis identified the pathological tumor size, lymph node metastasis, and Ki-67 expression as independent prognostic factors, which were entered into the nomogram model. The nomogram-predicted probabilities of OS at 1 year, 3 years, and 5 years posttreatment represented optimal concordance with the actual observations. Harrell's C-index of the constructed nomogram with the training set was 0.856 (95% CI: 0.804-0.908), whereas TNM staging was 0.814 (95% CI: 0.742-0.886, P = 5.280221e - 13). Survival analysis demonstrated that NSCLC subgroups showed significant differences in the training and validation sets (P < 0.001). A nomogram model was established for predicting survival in NSCLC patients with a pathological tumor size less than 30 mm, which would be further validated using demographic and clinicopathological data. In the future, this prognostic model may assist clinicians during treatment planning and clinical studies.

Prognostic value of mutant p53, Ki-67, and TTF-1 and their correlation with EGFR mutation in patients with non-small cell lung cancer.

INTRODUCTION: The clinical characteristics of non-small cell lung cancer (NSCLC) with epidermal growth factor receptor (EGFR) mutation have been well studied. However, the correlation of EGFR mutation with mutant p53, Ki-67, and thyroid transcription factor 1 (TTF-1) and their prognostic value remain indistinct. MATERIAL AND METHODS: Clinical and pathological characteristics and overall survival were analysed retrospectively in 523 surgically resected NSCLC patients. The expression levels of p53, Ki-67, and TTF-1 protein were detected by immunohistochemistry, and an amplification refractory mutation system was used to access the status of EGFR mutations. RESULTS: Of 523 patients with surgically resected NSCLC, 210 patients (38.4%) harboured EGFR mutations. Compared to the EGFR wild-type lung cancer, mutated EGFR harboured significantly increased mutant p53-positive or TTF-1-positive tumors (P<0.001 and<0.001, respectively). Former or current smokers, pathological stage and mutant p53-or TTF-1-positive status were independent predictors of EGFR mutation (P=0.001, 0.014, 0.014 and <0.001, respectively). Patients with p53 under expression had significantly better overall survival in the whole cohort and wild-type EGFR cohort (P=0.0010 and 0.0020, respectively) as well as in Ki-67-negative and TTF-1-positive patients (P<0.0001 and 0.0009, and P<0.0001 and 0.0004, respectively). Interestingly, in patients harbouring EGFR mutations, p53-under expression and Ki-67-negative cases still had better survival than positive cases, whereas there was no obvious difference between TTF-1-negative and TTF-1-positive cases (P=0.0198, 0.0068 and 0.3684, respectively). Finally, in NSCLC patients with wild-type EGFR, positive Ki-67 expression was the independent predictor for the worst survival (P=0.022). CONCLUSION: The expression levels of mutant p53, Ki-67, and TTF-1 were correlated with EGFR mutation. High expression of mutant p53 and Ki-67 correlated with poor survival in the entire cohort, EGFR mutation or wild-type cohort. In addition, Ki-67 might have an impact on the prognosis for patients with NSCLC with wild-type EGFR.

Iron-rich microorganism-enabled synthesis of magnetic biocarbon for efficient adsorption of diclofenac from aqueous solution.

Microorganisms in nature have been suggested as effective synthetic platform for functional materials construction. In this study, we cultured a typical white rot fungus of Phanerochaete chrysosporium in iron-containing medium to obtain iron-rich biomass, serving as sole precursor for magnetic biocarbon synthesis. The accumulated iron in biomass reached to 4.6 wt%. After carbonization and activation, microporous magnetic biocarbon (Fe/BC) with high specific surface area of 1986 m2 g-1 was obtained. When applied as adsorbent for a model pharmaceutical (diclofenac sodium, DCF) removal from aqueous solution, a high adsorption capacity of 361.25 mg g-1 was found for the developed Fe/BC. Systematic isotherm, kinetic, thermodynamic and recycle studies were conducted to investigate adsorption behaviors of DCF onto Fe/BC. This work not only provides a novel strategy for magnetic biocarbon construction, but also envisions new perspective on the utilization of a variety of microorganisms in nature for functional materials preparation.

Investigation on the Interaction of Dabrafenib with Human Serum Albumin Using Combined Experiment and Molecular Dynamics Simulation: Exploring the Binding Mechanism, Esterase-like Activity, and Antioxidant Activity.

Dabrafenib is a novel targeted antimelanoma drug. The present work explored the binding mechanism of dabrafenib-human serum albumin (HSA) and the effect on the esterase-like activity and antioxidant activity of HSA by using 19F NMR, spectroscopy methods, and molecular dynamics simulation. The results of 19F NMR, fluorescence, and time-resolved fluorescence spectroscopy revealed that dabrafenib spontaneously binds to the subdomain IIIA of the HSA by hydrophobic action and forms a static complex. The binding affects the esterase-like activity of HSA but not its antioxidant activity. According to the results of molecular dynamics simulation, dabrafenib interacts with Arg410 and Tyr411, which are the key residue associated with the esterase-like activity of HSA. Meanwhile, dabrafenib does not interact with Cys34, the key residue associated with the antioxidant activity of HSA. The results of circular dichroism spectroscopy and molecular dynamics simulation show that the conformation of HSA is not affected by the binding of dabrafenib. This study can provide useful information for understanding the pharmacokinetic properties of dabrafenib.