Nanoenabled Intracellular Metal Ion Homeostasis Regulation for Tumor Therapy.

Endogenous essential metal ions play an important role in many life processes, especially in tumor development and immune response. The approval of various metallodrugs for tumor therapy brings more attention to the antitumor effect of metal ions. With the deepening understanding of the regulation mechanisms of metal ion homeostasis in vivo, breaking intracellular metal ion homeostasis becomes a new means to inhibit the proliferation of tumor cells and activate antitumor immune response. Diverse nanomedicines with the loading of small molecular ion regulators or metal ions have been developed to disrupt metal ion homeostasis in tumor cells, with higher safety and efficiency than free small molecular ion regulators or metal compounds. This comprehensive review focuses on the latest progress of various intracellular metal ion homeostasis regulation-based nanomedicines in tumor therapy including calcium ion (Ca2+ ), ferrous ion (Fe2+ ), cuprous ion (Cu+ ), managanese ion (Mn2+ ), and zinc ion (Zn2+ ). The physiological functions and homeostasis regulation processes of ions are summarized to guide the design of metal ion regulation-based nanomedicines. Then the antitumor mechanisms of various ions-based nanomedicines and some efficient synergistic therapies are highlighted. Finally, the challenges and future developments of ion regulation-based antitumor therapy are also discussed, hoping to provide a reference for finding more effective metal ions and synergistic therapies.

The adcA and lmb Genes Play an Important Role in Drug Resistance and Full Virulence of Streptococcus suis.

Streptococcus suis is an recognized zoonotic pathogen of swine and severely threatens human health. Zinc is the second most abundant transition metal in biological systems. Here, we investigated the contribution of zinc to the drug resistance and pathogenesis of S. suis. We knocked out the genes of AdcACB and Lmb, two Zn-binding lipoproteins. Compared to the wild-type strain, we found that the survival rate of this double-mutant strain (ΔadcAΔlmb) was reduced in Zinc-limited medium, but not in Zinc-supplemented medium. Additionally, phenotypic experiments showed that the ΔadcAΔlmb strain displayed impaired adhesion to and invasion of cells, biofilm formation, and tolerance of cell envelope-targeting antibiotics. In a murine infection model, deletion of the adcA and lmb genes in S. suis resulted in a significant decrease in strain virulence, including survival rate, tissue bacterial load, inflammatory cytokine levels, and histopathological damage. These findings show that AdcA and Lmb are important for biofilm formation, drug resistance, and virulence in S. suis. IMPORTANCE Transition metals are important micronutrients for bacterial growth. Zn is necessary for the catalytic activity and structural integrity of various metalloproteins involved in bacterial pathogenic processes. However, how these invaders adapt to host-imposed metal starvation and overcome nutritional immunity remains unknown. Thus, pathogenic bacteria must acquire Zn during infection in order to successfully survive and multiply. The host uses nutritional immunity to limit the uptake of Zn by the invading bacteria. The bacterium uses a set of high-affinity Zn uptake systems to overcome this host metal restriction. Here, we identified two Zn uptake transporters in S. suis, AdcA and Lmb, by bioinformatics analysis and found that an adcA and lmb double-mutant strain could not grow in Zn-deficient medium and was more sensitive to cell envelope-targeting antibiotics. It is worth noting that the Zn uptake system is essential for biofilm formation, drug resistance, and virulence in S. suis. The Zn uptake system is expected to be a target for the development of novel antimicrobial therapies.

Metabolomics and proteomics analyses revealed mechanistic insights on the antimicrobial activity of epigallocatechin gallate against Streptococcus suis.

Streptococcus suis (S. suis) is a highly virulent zoonotic pathogen and causes severe economic losses to the swine industry worldwide. Public health security is also threatened by the rapidly growing antimicrobial resistance in S. suis. Therefore, there is an urgent need to develop new and safe antibacterial alternatives against S. suis. The green tea polyphenol epigallocatechin gallate (EGCG) with a number of potential health benefits is known for its antibacterial effect; however, the mechanism of its bactericidal action remains unclear. In the present, EGCG at minimal inhibitory concentration (MIC) showed significant inhibitory effects on S. suis growth, hemolytic activity, and biofilm formation, and caused damage to S. suis cells in vitro. EGCG also reduced S. suis pathogenicity in Galleria mellonella larvae in vivo. Metabolomics and proteomics analyses were performed to investigate the underlying mechanism of antibacterial activity of EGCG at MIC. Many differentially expressed proteins involved in DNA replication, synthesis of cell wall, and cell membrane, and virulence were down-regulated after the treatment of S. suis with EGCG. EGCG not only significantly reduced the hemolytic activity of S. suis but also down-regulated the expression of suilysin (Sly). The top three shared KEGG pathways between metabolomics and proteomics analysis were ABC transporters, glycolysis/gluconeogenesis, and aminoacyl-tRNA biosynthesis. Taken together, these data suggest that EGCG could be a potential phytochemical compound for treating S. suis infection.

The VraSR two-component signal transduction system contributes to the damage of blood-brain barrier during Streptococcus suis meningitis.

Streptococcus suis (S. suis) is an important zoonotic pathogen that can cause high morbidity and mortality in both humans and swine. As the most important life-threatening infection of the central nervous system (CNS), meningitis is an important syndrome of S. suis infection. The vancomycin resistance associated sensor/regulator (VraSR) is a critical two-component signal transduction system that affects the ability of S. suis to resist the host innate immune system and promotes its ability to adhere to brain microvascular endothelial cells (BMECs). Prior work also found mice infected with ΔvraSR had no obvious neurological symptoms, unlike mice infected with wild-type SC19. Whether and how VraSR participates in the development of S. suis meningitis remains unknown. Here, we found ΔvraSR-infected mice did not show obvious meningitis, compared with wild-type SC19-infected mice. Moreover, the proinflammatory cytokines and chemokines in serum and brains of ΔvraSR-infected mice, including IL-6, TNF-α, MCP-1 and IFN-γ, were significantly lower than wild-type infected group. Besides, blood-brain barrier (BBB) permeability also confirmed that the mutant had lower ability to disrupt BBB. Furthermore, in vivo and in vitro experiments showed that SC19 could increase BBB permeability by downregulating tight junction (TJ) proteins such as ZO-1, ß-Catenin, Occludin, and Clauidn-5, compared with mutant ΔvraSR. These findings provide new insight into the influence of S. suis VraSR on BBB disruption during the pathogenic process of streptococcal meningitis, thereby offering potential targets for future preventative and therapeutic strategies against this disease.

[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.

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.

IL-35 ameliorates collagen-induced arthritis by promoting TNF-α-induced apoptosis of synovial fibroblasts and stimulating M2 macrophages polarization.

Synovitis, the chronic inflammation of the synovial membranes, is a hallmark of rheumatoid arthritis, a chronic disease with profound impact on human health. Recently, interleukin-35 (IL-35), a new member of the IL-12 family, was identified as an anti-inflammatory and immunosuppressive cytokine and was shown to ameliorate collagen-induced arthritis (CIA) in mice. However, the mechanism by which IL-35 alleviates CIA remains unknown. In this study, we investigated the effect of IL-35 on the CIA microenvironment and, specifically, the tumor necrosis factor alpha (TNF-α)-induced macrophage inflammatory response and apoptosis of fibroblast-like synoviocytes (FLSs). Firstly, using RT-PCR, western blot, and flow cytometry, we found that IL-35 suppressed TNF-α-induced inflammatory responses by down-regulating iNOS and COX-2 in peripheral blood monocyte-derived macrophages. IL-35 also activated alternative M2 macrophage polarization, as determined by evaluation of CCR7 and CD206 expression. Moreover, we showed that IL-35 enhanced TNF-α-induced FLS apoptosis. Using a panel of immunohistochemical and immunofluorescence analyses in a CIA model established in 18 DBA/1J mice, we demonstrated that IL-35 promotes synoviocyte apoptosis and alternative activation of macrophages to alleviate arthritis in vivo. Taken together, our results show that IL-35 promotes TNF-α-induced FLS apoptosis and modulates M2 macrophage polarization to ameliorate CIA inflammation both in vitro and in vivo.

Correction: Preoperative Prediction of Ki-67 Labeling Index By Three-dimensional CT Image Parameters for Differential Diagnosis Of Ground-Glass Opacity (GGO).

[This corrects the article DOI: 10.1371/journal.pone.0129206.].

Inhibition of JNK and activation of the AMPK-Nrf2 axis by corosolic acid suppress osteolysis and oxidative stress.

The intracellular reactive oxygen species contribute to RANKL-induced osteoclastogenesis and osteolysis. Nuclear factor-erythroid 2-related factor 2 (Nrf2), a redox-sensitive transcription factor, is critical in the cellular defense against oxidative stress by induction of antioxidants and cytoprotective enzymes. In the current study, it was first demonstrated that RANKL-induced osteoclastogenesis and hydroxylapatite resorption were suppressed by Corosolic acid (CA) via inhibiting p-JNK and activating p-AMPK. Meanwhile, p-65, p-38, Akt, and GSK-3ß were partly inhibited during the treatment of CA. Osteoclastogenesis related genes, including NFATc1, c-fos, cathepsin K, and CTR were down-regulated by CA as well. Furthermore, the intracellular oxidative stress of CA-treated osteoclasts was dramatically decreased and Nrf2 was translocated into the nucleus to activate antioxidants including HO-1, NQO-1, and GCLC by CA. The LPS-induced mice calvarial osteolysis model was established for the in vivo investigation. Micro-CT morphometric analysis revealed that the treatment of CA restored LPS-induced bone loss and formation of osteoclasts. Besides, p-p65 and p-JNK were activated in the LPS group but inhibited by CA in vivo. The treatment of CA also activated p-AMPK during its attenuating LPS-induced osteolysis. Conclusively, CA effectively protects against LPS-induced osteolysis by suppressing osteoclastogenesis and oxidative stress through the inhibition of the JNK and activation of the AMPK-Nrf2 axis.

Controllable fabrication of hydroxybutyl chitosan/oxidized chondroitin sulfate hydrogels by 3D bioprinting technique for cartilage tissue engineering.

Biological regeneration of articular cartilage continues to be a challenge at present. Functional engineered implants with patient-specific sizes are difficult to achieve. The aim of this study is to fabricate a biocompatible cell-laden hydrogel with a designable structure. Covalent hydrogels were prepared with water soluble hydroxybutyl chitosan (HBC) and oxidized chondroitin sulfate (OCS) via a Schiff-base reaction. With the aid of three-dimensional (3D) bioprinted sacrificial molds, HBC/OCS hydrogel with various structures were obtained. After the material constituent optimization process, an injectable hydrogel with a uniform porous structure of 100 µm average pore size was developed to form macroporous hydrogel. In vitro and in vivo biocompatibility of optimized HBC/OCS hydrogel were also carefully assessed. The results indicated that human adipose-derived mesenchymal stem cells could be 3D cultured in HBC/OCS hydrogel maintaining good viability. Moreover, the hydrogels were found to trigger the least amount of pro-inflammatory gene expression of macrophage and to inhibit acute immune responses in 7 d. These results demonstrate the potential of HBC/OCS hydrogels as a cell delivery system for cartilage tissue engineering.

Modification of Cysteine 179 in IKKß by Ursolic Acid Inhibits Titanium-Wear-Particle-Induced Inflammation, Osteoclastogenesis, and Hydroxylapatite Resorption.

Aseptic loosening of artificial joints mainly accounts for the failure of arthroplasty. We previously reported that ursolic acid (UA) inhibited osteolysis caused by titanium (Ti) wear particles via suppression of NF-kB signaling. In the present study, that the suppressive effect of UA on Ti-particle-induced inflammation and osteoclastogenesis targets on IKKß cys-179 was demonstrated. A retrovirus packaged IKKßC179A plasmid with a Cys-179 mutation replaced by Ala was constructed. qRT-PCR, immunoblot, and immunofluorescence were used to evaluate the gene expressions. Secreted inflammatory cytokines were detected by ELISA. Formation and function of osteoclastogenesis were evaluated by TRAP stain and hydroxylapatite resorption assays. As a result, a mutation of IKKßC179A rescued the therapeutic effect of UA on Ti-particle-induced inflammation, including morphological transforms, upregulation of iNOS and COX-2, increased secretions of TNF-α, IL-1ß, and IL-6, and decreased secretion of IL-10. Meanwhile, inhibition of osteoclastogenesis and hydroxylapatite resorptions were restored by transfection of IKKßC179A. Phosphorylations of p65 and the IKKα/ß complex and translocation of p65 into the nucleus were suppressed by UA but rescued by a mutation of IKKßC179A. Conclusively, UA inhibits Ti-wear-particle-induced inflammation, osteoclastogenesis, and hydroxylapatite resorption via modifying cysteine 179 of IKKß.

Interleukin-35 Inhibits TNF-α-Induced Osteoclastogenesis and Promotes Apoptosis via Shifting the Activation From TNF Receptor-Associated Death Domain (TRADD)-TRAF2 to TRADD-Fas-Associated Death Domain by JAK1/STAT1.

Over-activated osteoclasts derived from myeloid or peripheral blood monocytes by inflammatory cytokines results in osteoporosis, osteoarthritis, and other bone erosion-related diseases. Interleukin 35 (IL-35) is a novel anti-inflammatory and immunosuppressive factor. This study investigated the effect of IL-35 on TNF-α-induced osteoclastogenesis. In the presence of IL-35, this process was detected by Tartrate-Resistant Acid Phosphatase (TRAP) staining, F-actin staining, and bone resorption assays. The effects of IL-35 on TNF-α-induced apoptosis were demonstrated by TUNEL staining, cell viability assays, and flow cytometry. Moreover, a microarray was performed to detect the effect of IL-35 on TNF-α-activated phosphatase kinase. The effect of IL-35 on the TNF-α-mediated activation of NF-κB, MAPK, TRAF2, RIP1, Fas-associated death domain (FADD), and caspase3 was further investigated. In addition, a murine calvarial osteolysis model was established via the subcutaneous injection of TNF-α onto the calvaria, and histological analysis was subsequently performed. As a result, IL-35 inhibited TNF-α-induced osteoclast formation and bone resorption in vitro and osteolysis calvaria in vivo. NFATc1, c-fos, and TRAP were downregulated by IL-35 through the inhibition of NF-κB and MAPK, during which JAK1/STAT1 was activated. Moreover, based on TUNEL staining and flow cytometry, IL-35 was shown to enhance TNF-α-induced osteoclast apoptosis. Meanwhile, FADD and cleaved-caspase 3 were increased in cells treated with TNF-α and IL-35, whereas the DNA-binding activity of NF-κB was increased in TNF-α-treated cells, but was decreased in cells treated with both TNF-α and IL-35. In conclusion, IL-35 inhibits TNF-α-induced osteoclastogenesis and promotes apoptosis by activating JAK1/STAT1 and shifting activation from TNF receptor-associated death domain (TRADD)-TRAF2/RIP1-NF-κB to TRADD-FADD-caspase 3 signaling.

3D-printed IFN-γ-loading calcium silicate-ß-tricalcium phosphate scaffold sequentially activates M1 and M2 polarization of macrophages to promote vascularization of tissue engineering bone.

To promote vascularization of tissue-engineered bone, IFN-γ polarizing macrophages to M1 was loaded on 5% calcium silicate/ß-tricalcium phosphate (CaSiO3-ß-TCP) scaffolds. IFN-γ and Si released from the scaffold were designed to polarize M1 and M2 macrophages, respectively. ß-TCP, CaSiO3-ß-TCP, and IFN-γ@CaSiO3-ß-TCP were fabricated and biocompatibilities were evaluated. Polarizations of macrophages were detected by flow cytometry. Human umbilical vein endothelial cells with GFP were cultured and induced on Matrigel with conditioned culture medium extracted from culture of macrophages loaded on scaffolds for evaluating angiogenesis. Four weeks after the scaffolds were subcutaneously implanted into C57B1/6, vascularization was evaluated by visual observation, hematoxylin and eosin staining, as well as immunohistochemistry of CD31. The results showed that IFN-γ@CaSiO3-ß-TCP scaffolds released IFN-γ in the early stage (1-3 days) to stimulate macrophages to M1 polarization, followed by release of Si inducing macrophages to M2 polarization while scaffolds degraded. The activation of M1/M2 allows macrophages to secrete more cytokines, including VEGF, CXCL12 and PDGF-BB. The IFN-γ@CaSiO3-ß-TCP scaffolds formed more blood vessels in vitro and in vivo compared to the control groups. The study indicated that the design of tissue-engineered scaffolds with immunomodulatory function utilized host macrophages to increase vascularization of tissue-engineered bone, providing a new strategy for accelerating vascularization and osteogenesis of tissue-engineered scaffolds and showing the potential for treatment of major bone defects. STATEMENT OF SIGNIFICANCE: A 3-D printed immunomodulatory scaffold was designed for repair of massive bone defects. Through the release of interferon γ and silicon ions, the new immunomodulatory scaffold promoted the M1 and M2 polarization of macrophages, boosting angiogenesis. This scaffold provided a new strategy for accelerating vascularization and osteogenesis of tissue-engineered scaffolds and showing the potential for treatment of major bone defects.

Proinflammatory and osteolysis-inducing effects of 3D printing Ti6Al4V particles in vitro and in vivo.

Ti6Al4V printing particles have been recently used for fabricating orthopedic implants. Removing these particles completely from fabricated implants is challenging. Furthermore, recycled particles are commonly used in fabrication without additional analysis. Ti6Al4V wear particles derived from orthopedic implants are known to induce inflammatory responses and osteolysis. However, the biosafety of printing particles remains unknown. Here, we investigated the proinflammatory and osteolysis-inducing effects of commonly used original and recycled Ti6Al4V printing particles in vitro and in vivo. Our results indicated that although less serious effects were induced compared to wear particles, inflammatory responses and osteoclast-mediated bone resorption were induced by the original printing particles in a particle size-dependent manner. Recycled particles were found to more strongly stimulate bone resorption and inflammatory responses than the original particles; the in vivo effect was enhanced with an increase in particle concentration. Furthermore, the results of our in vitro experiments verified that the printing particles activate macrophages to secrete inflammatory cytokines and promote osteoclastogenesis, which is closely related to particle size and concentration. Taken together, our findings provide a valuable reference for the use of raw printing materials and examination of recycling procedures for implant fabrication.

3D printed scaffolds of calcium silicate-doped ß-TCP synergize with co-cultured endothelial and stromal cells to promote vascularization and bone formation.

Synthetic bone scaffolds have potential application in repairing large bone defects, however, inefficient vascularization after implantation remains the major issue of graft failure. Herein, porous ß-tricalcium phosphate (ß-TCP) scaffolds with calcium silicate (CS) were 3D printed, and pre-seeded with co-cultured human umbilical cord vein endothelial cells (HUVECs) and human bone marrow stromal cells (hBMSCs) to construct tissue engineering scaffolds with accelerated vascularization and better bone formation. Results showed that in vitro ß-TCP scaffolds doped with 5% CS (5%CS/ß-TCP) were biocompatible, and stimulated angiogenesis and osteogenesis. The results also showed that 5%CS/ß-TCP scaffolds not only stimulated co-cultured cells angiogenesis on Matrigel, but also stimulated co-cultured cells to form microcapillary-like structures on scaffolds, and promoted migration of BMSCs by stimulating co-cultured cells to secrete PDGF-BB and CXCL12 into the surrounding environment. Moreover, 5%CS/ß-TCP scaffolds enhanced vascularization and osteoinduction in comparison with ß-TCP, and synergized with co-cultured cells to further increase early vessel formation, which was accompanied by earlier and better ectopic bone formation when implanted subcutaneously in nude mice. Thus, our findings suggest that porous 5%CS/ß-TCP scaffolds seeded with co-cultured cells provide new strategy for accelerating tissue engineering scaffolds vascularization and osteogenesis, and show potential as treatment for large bone defects.

Three-dimensional substructure measurements for the differential diagnosis of ground glass nodules.

BACKGROUND: We analyzed the differences between maximum and peak computed tomography (CT) numbers (M-P), respectively representing the densities of the solid center and the main periphery of ground-glass nodules (GGNs), and the average change in M-P velocity (V(M-P)) during follow-up to differentiate between pre-invasive (PIA) and invasive adenocarcinoma (IAC). METHODS: Data of 102 patients were retrospectively collected and analyzed in our study including 43 PIAs and 59 IACs. Diameters, total volumes, and the maximum and peak CT numbers in CT number histograms were measured and followed for at least 3 months. This study was registered retrospectively. RESULTS: The M-P values for IACs were higher than those for PIAs (p = 0.001), with an area under the curve (AUC) of 0.810 and a threshold of 489.5 Hounsfield units (HU) in ROC analysis. The V(M-P) values for IACs were smaller than those for PIAs (p = 0.04), with an AUC of 0.805 and a threshold of 11.01 HU/day. CONCLUSIONS: M-P and V(M-P) values may help distinguish IACs from PIAs by representing the changes in the sub-structural densities of GGNs during follow-up.

RhBMP-2 loaded 3D-printed mesoporous silica/calcium phosphate cement porous scaffolds with enhanced vascularization and osteogenesis properties.

A major limitation in the development of effective scaffolds for bone regeneration has been the limited vascularization of the regenerating tissue. Here, we propose the development of a novel calcium phosphate cement (CPC)-based scaffold combining the properties of mesoporous silica (MS) with recombinant human bone morphogenic protein-2 (rhBMP-2) to facilitate vascularization and osteogenesis. Specifically, the development of a custom MS/CPC paste allowed the three-dimensional (3D) printing of scaffolds with a defined macroporous structure and optimized silicon (Si) ions release profile to promote the ingrowth of vascular tissue at an early stage after implantation in support of tissue viability and osteogenesis. In addition, the scaffold microstructure allowed the prolonged release of rhBMP-2, which in turn significantly stimulated the osteogenesis of human bone marrow stromal cells in vitro and of bone regeneration in vivo as shown in a rabbit femur defect repair model. Thus, the combination MS/CPC/rhBMP-2 scaffolds might provide a solution to issues of tissue necrosis during the regeneration process and therefore might be able to be readily developed into a useful tool for bone repair in the clinic.

Pulmonary ground-glass nodules diagnosis: mean change rate of peak CT number as a discriminative factor of pathology during a follow-up.

OBJECTIVE: We aimed to analyse the peak CT number (PEAK) in CT number histogram of ground-glass nodules (GGN), meaning the most frequent density of pixels in the image of pulmonary nodule, based on three-dimensional (3D) reconstructive model pre-operatively, and the mean rate of PEAK change (V-PEAK) during a follow-up of GGN for differential diagnosis between pre-invasive adenocarcinoma (PIA) and invasive adenocarcinoma (IAC). METHODS: CT number histogram of pixels in GGN was made automatically by 3D measurement software. Diameter, total volume, PEAK and V-PEAK were measured from CT data sets of different groups classified by pathology, subtype and number of GGN, respectively. RESULTS: Among all 102 cases, 47 were PIA, including atypical adenomatous hyperplasia (n = 29) and adenocarcinoma in situ (n = 18), and 55 were IAC, including minimally IAC (MIA, n = 4). By Wilcoxon test, PEAK of IAC was significantly higher than that of PIA (p < 0.001). By receiver operating curve analysis, area under the curve (AUC) was 0.857 and threshold -820.50 Hounsfield units (HU) for differentiation between PIA and IAC. V-PEAK of IAC was unexpectedly remarkably smaller than that of PIA (p < 0.001) with AUC and threshold being 0.810 and -0.829 HU day(-1), respectively. CONCLUSION: Pre-operative PEAK and V-PEAK, which interpret and evaluate the change of volume and density of pulmonary nodule simultaneously from both exterior and interior perspectives, can help to distinguish IAC from PIA. ADVANCES IN KNOWLEDGE: This study provided researchers of GGN another perspective, taking both volume and density of nodules into consideration for pathological evaluation.

Three-dimensional vs two-dimensional video assisted thoracoscopic esophagectomy for patients with esophageal cancer.

AIM: To define the benefits of three-dimensional video-assisted thoracoscopic esophagectomy (3D-VATE) over 2D-VATE for esophageal cancer. METHODS: A total of 93 patients with esophageal cancer including 45 patients receiving 3D-VATE and 48 receiving 2D-VATE were evaluated. Data related to patient and cancer characteristics, operating time, intraoperative bleeding, morbidity and mortality, postoperative inflammatory markers, Numerical Rating Scale for postoperative pain, Constant-Murley rating system for shoulder recovery and oxygenation index (OI) were collected. All medical records were retrieved from a prospectively maintained oncological database at our institution. A retrospective study was performed to compare the short-term surgical outcomes between the two groups. RESULTS: No significant differences were found between the two groups in either morbidity or mortality (P = 0.328). An enhanced surgical recovery was noted in the 3D group as indicated by shortened thoracoscopic operation time (3D vs 2D: 68 ± 13.79 min vs 83 ± 13 min, P < 0.01), minor intraoperative blood loss (3D vs 2D: 68.2 ± 10.7 mL vs 89.8 ± 10.4 mL, P < 0.01), earlier chest tube removal (3D vs 2D: 2.67 ± 1.01 vs 3.75 ± 1.15 d, P < 0.01), shorter length of hospital stay (3D vs 2D: 9.07 ± 2.00 vs 10.85 ± 3.40 d, P < 0.01), lower in-hospital expenses (3D vs 2D: 74968.4 ± 9637.8 vs 86211.1 ± 8519.7 RMB, P < 0.01), lower pain intensity (P < 0.01) and faster recovery of the left shoulder function (P < 0.01). Better preservation of the pulmonary function was also found in the 3D group as the decline of the OI post operation was significantly lower than that of the 2D group (P < 0.01). Changes of postoperative inflammatory markers, including procalcitonin [postoperative days (PODs) 4 and 7: P < 0.01], peripheral granulocytes (PODs 1, 4 and 7: P < 0.01) and hypersensitive C-reactive protein (POD 4: P < 0.01) in 3D-VATE patients were less than those in the 2D group. Moreover, utilization of the 3D technique extended the dissection of the thoracic lymph nodes (P < 0.01), with better exposure of nodes in the left recurrent laryngeal nerve (P = 0.031). CONCLUSION: 3D-VATE could be a more viable technique over 2D-VATE in terms of short-term outcomes for patients with esophageal cancer.

Preoperative Prediction of Ki-67 Labeling Index By Three-dimensional CT Image Parameters for Differential Diagnosis Of Ground-Glass Opacity (GGO).

The aim of this study was to predict Ki-67 labeling index (LI) preoperatively by three-dimensional (3D) CT image parameters for pathologic assessment of GGO nodules. Diameter, total volume (TV), the maximum CT number (MAX), average CT number (AVG) and standard deviation of CT number within the whole GGO nodule (STD) were measured by 3D CT workstation. By detection of immunohistochemistry and Image Software Pro Plus 6.0, different Ki-67 LI were measured and statistically analyzed among preinvasive adenocarcinoma (PIA), minimally invasive adenocarcinoma (MIA) and invasive adenocarcinoma (IAC). Receiver operating characteristic (ROC) curve, Spearman correlation analysis and multiple linear regression analysis with cross-validation were performed to further research a quantitative correlation between Ki-67 labeling index and radiological parameters. Diameter, TV, MAX, AVG and STD increased along with PIA, MIA and IAC significantly and consecutively. In the multiple linear regression model by a stepwise way, we obtained an equation: prediction of Ki-67 LI=0.022*STD+0.001* TV+2.137 (R=0.595, R's square=0.354, p<0.001), which can predict Ki-67 LI as a proliferative marker preoperatively. Diameter, TV, MAX, AVG and STD could discriminate pathologic categories of GGO nodules significantly. Ki-67 LI of early lung adenocarcinoma presenting GGO can be predicted by radiologic parameters based on 3D CT for differential diagnosis.