Collective transport and reconfigurable assembly of nematic colloids by light-driven cooperative molecular reorientations.

Nanomotors in nature have inspired scientists to design synthetic molecular motors to drive the motion of microscale objects by cooperative action. Light-driven molecular motors have been synthesized, but using their cooperative reorganization to control the collective transport of colloids and to realize the reconfiguration of colloidal assembly remains a challenge. In this work, topological vortices are imprinted in the monolayers of azobenzene molecules which further interface with nematic liquid crystals (LCs). The light-driven cooperative reorientations of the azobenzene molecules induce the collective motion of LC molecules and thus the spatiotemporal evolutions of the nematic disclination networks which are defined by the controlled patterns of vortices. Continuum simulations provide physical insight into the morphology change of the disclination networks. When microcolloids are dispersed in the LC medium, the colloidal assembly is not only transported and reconfigured by the collective change of the disclination lines but also controlled by the elastic energy landscape defined by the predesigned orientational patterns. The collective transport and reconfiguration of colloidal assemblies can also be programmed by manipulating the irradiated polarization. This work opens opportunities to design programmable colloidal machines and smart composite materials.

Active transformations of topological structures in light-driven nematic disclination networks.

SignificanceTopological defects are marvels of nature. Understanding their structures is important for their applications in, for example, directed self-assembly, sensing, and photonic devices. There is recent interest in active motion and transformation of topological defects in active nematics. In these nonequilibrium systems, however, the motion and transformation of disclinations are difficult to control, thereby hindering their applications. Here, we propose a surface-patterned system engendering periodic three-dimensional disclinations, which can be excited by light irradiation and undergo a programmable transformation between different topological states. Continuum simulations recapitulating these topological structures characterize the bending, breaking, and relinking events of the disclinations during the nonequilibrium process. Our work provides an alternative dynamic system in which active transformation of topological defects can be engineered.

AIF1 + CSF1R + MSCs, induced by TNF-α, act to generate an inflammatory microenvironment and promote hepatocarcinogenesis.

BACKGROUND AND AIMS: Increasing evidence suggests that mesenchymal stem cells (MSCs) home to injured local tissues and the tumor microenvironment in the liver. Chronic inflammation is regarded as the major trait of primary liver cancer. However, the characteristics of endogenous MSCs in the inflammatory environment and their role in the occurrence of liver cancer remain obscure. APPROACH AND RESULTS: Using single-cell RNA sequencing, we identified a distinct inflammation-associated subset of MSCs, namely AIF1 + CSF1R + MSCs, which existed in the microenvironment before the occurrence of liver cancer. Furthermore, we found that this MSC subgroup is likely to be induced by TNF-α stimulation through the TNFR1/SIRT1 (sirtuin 1) pathway. In a rat primary liver cancer model, we showed that MSCs with high SIRT1 expression (Ad-Sirt1-MSCs) promoted macrophage recruitment and synergistically facilitated liver cancer occurrence by secreting C-C motif chemokine ligand (CCL) 5. Interestingly, depletion of macrophages or knockdown of CCL5 expression in Ad-Sirt1-MSCs attenuated the promotive effect of Ad-Sirt1-MSCs on liver inflammation and hepatocarcinogenesis (HCG). Finally, we demonstrated that SIRT1 up-regulated CCL5 expression through activation of the AKT/HIF1α signaling axis in MSCs. CONCLUSIONS: Together, our results show that MSCs, which are mobilized to the injured site, can be educated by macrophages. In turn, the educated MSCs are involved in generating a chronic inflammatory microenvironment and promoting HCG.

miR-671-5p Promotes Cell Proliferation, Invasion, and Migration in Hepatocellular Carcinoma through Targeting ALDH2.

We explored the mechanism of acetaldehyde dehydrogenase 2 (ALDH2) in modulating cell behaviors in hepatocellular carcinoma (HCC), and provided fresh ideas for targeted treatment of HCC. The target messenger RNA (mRNA) was determined by The Cancer Genome Atlas (TCGA) analysis, and the upstream regulatory gene miRNA was obtained by further analysis. The expression of ALDH2 mRNA and miR-671-5p in HCC cell lines was assayed by quantitative reverse transcription polymerase chain reaction (qRT-PCR), and protein expression was assessed by Western blot. The impact of ALDH2 on biological functions of HCC cells was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), wound-healing, and Transwell assays. Bioinformatics method was utilized to predict binding site of miR-671-5p and ALDH2, and their targeted relationship was detected by dual-luciferase gene assay, qRT-PCR and Western blot. ALDH2 expression was reduced in HCC tissue and cell lines. ALDH2 worked as a tumor inhibitor in HCC. Overexpressing ALDH2 could hinder proliferation, migration and invasion of HCC cells. miR-671-5p was the upstream regulatory gene of ALDH2, and it presented remarkably high expression in HCC. A negative modulatory relationship existed between miR-671-5p and ALDH2. The rescue experiments further illustrated the effects of the two on the malignant behaviors of HCC cells. Forced expression of miR-671-5p fostered the proliferation, migration and invasion of HCC cells by restraining ALDH2.

Targeted blocking of CCR2 and CXCR2 improves the efficacy of transarterial chemoembolization of hepatocarcinoma.

BACKGROUND: Transarterial chemoembolization (TACE) has been shown to prolong survival in patients with unresectable hepatocellular carcinoma (HCC); however, the long-term survival remains dismal. Targeting macrophage and neutrophil infiltration is a promising strategy. The CCL2/CCR2 and CXCLs/CXCR2 axes are required for recruitment of macrophages and neutrophils, respectively, in HCC. We investigated the feasibility of CCL2/CCR2 and CXCLs/CXCR2 as therapeutic targets in combination with TACE for treating HCC. METHODS: Expression of CCL2/CCR2 and CXCLs/CXCR2 was analyzed in the primary rat HCC model and one HCC cohort. The relationship between expression levels, neutrophil and macrophage infiltration, hepatocarcinogenesis progression in the rat model, and survival of HCC patients was assessed. The anti-tumor effects of blocking the CCL2/CCR2 and CXCLs/CXCR2 axes by CCR2 and CXCR2 antagonists in combination with TACE were evaluated in HCC rats. The numbers of macrophages, neutrophils, and hepatic progenitor cells were further determined to explore the underlying mechanisms. RESULTS: High macrophage and neutrophil infiltration and CXCL8 expression were associated with poor prognosis in the TCGA liver cancer dataset. High expression of CCL2/CCR2 and CXCL8/CXCR2 in clinical HCC specimens was associated with reduced survival. Expression of CCL2/CCR2 and CXCL1/CXCR2 was correlated with hepatocarcinogenesis progression in the primary rat HCC model. Blockade of CCL2/CCR2 and CXCLs/CXCR2 enhanced the anti-tumor effect of TACE treatment in this model. Blocking the CCL2/CCR2 and CXCLs/CXCR2 axes with CCR2 and CXCR2 antagonists in TACE-treated rats reduced macrophage and neutrophil infiltration and hepatic progenitor cell activation and thus overcame TACE resistance in HCC. CONCLUSIONS: The results demonstrate the translational potential of immunotherapy targeting the CCL2/CCR2 and CXCLs/CXCR2 axes in combination with TACE therapy for the treatment of HCC.

Sirt1-Overexpressing Mesenchymal Stem Cells Drive the Anti-tumor Effect through Their Pro-inflammatory Capacity.

The major obstacles for the efficacy of tumor immunotherapies are their immune-related systemic adverse events. Therefore, tumor tropism property and pro-inflammatory ability of mesenchymal stem cells (MSCs) could be utilized in combination to potentiate local immunity for cancer eradication. We previously observed that MSCs with the type III histone deacetylase silent information regulator 2 homologue 1 (Sirt1) overexpression displayed a pro-inflammatory capacity. However, the anti-tumor effect of Sirt1-overexpressing MSCs and the role of Sirt1 in regulating the pro-inflammatory capacity of MSCs still need to be clarified. In this study, utilizing the hepatic metastasis model of colorectal carcinoma, we demonstrated that Sirt1-overexpressing MSCs significantly exerted anti-tumor activity through increasing the number of CD8+ T cells. Furthermore, Sirt1 did not affect chemokine secretion in MSCs induced by inflammatory cytokines, but impaired the immunosuppressive ability of MSCs through suppressing inflammatory cytokine-stimulated inducible nitric oxide synthase (iNOS) production via deacetylating p65. iNOS overexpression negated the anti-tumor effect of Sirt1-overexpressing MSCs. Collectively, our data defined Sirt1 as the critical regulator for modulating the pro-inflammatory ability of MSCs, and they suggested that Sirt1-overexpressing MSCs secreting chemokines but little iNOS under the inflammatory milieu were capable of attracting immune cells to close proximity without suppressing their proliferation, thereby achieving a potent anti-tumor effect.

Autophagy and Tumour Chemotherapy.

Chemotherapy is an important means of treating malignant tumours. The main role of chemotherapy drugs is to induce cell death. However, the apoptotic pathways of many tumour cells are often severely impaired, leading to failure of chemotherapy-induced apoptosis. With the in-depth study of autophagy in recent years, this process has been found to play an important role in the chemoresistance of tumours. Autophagy may have different effects on tumour cells depending on the specific environment. In addition, tumour stem cells and the tumour microenvironment are closely related to tumour recurrence and metastasis. It is also important to study the role of autophagy in tumour stem cells and the microenvironment to investigate chemotherapy resistance.

Torin2 overcomes sorafenib resistance via suppressing mTORC2-AKT-BAD pathway in hepatocellular carcinoma cells.

BACKGROUND: Sorafenib is an oral multi-kinase inhibitor that was approved by the US Food and Drug Administration for the treatment of patients with advanced hepatocellular carcinoma (HCC). However, resistance to sorafenib is an urgent problem to be resolved to improve the therapeutic efficacy of sorafenib. As the activation of AKT/mTOR played a pivotal role in sorafenib resistance, we evaluated the effect of a dual mTOR complex 1/2 inhibitor Torin2 on overcoming the sorafenib resistance in HCC cells. METHODS: The sorafenib-resistant Huh7 and Hep3B cell lines were established from their parental cell lines. The synergistic effect of sorafenib and Torin2 on these cells was measured by cell viability assay and quantified using the Chou-Talalay method. Apoptosis induced by the combination of sorafenib and Torin2 and the alteration in the specific signaling pathways of interest were detected by Western blotting. RESULTS: Sorafenib treatment inversely inhibited AKT in parental but activated AKT in sorafenib-resistant Huh7 and Hep3B HCC cells, which underscores the significance of AKT activation. Torin2 and sorafenib synergistically suppressed the viability of sorafenib-resistant cells via apoptosis induction. Torin2 successfully suppressed the sorafenib-activated mTORC2-AKT axis, leading to the dephosphorylation of Ser136 in BAD protein, and increased the expression of total BAD, which contributed to the apoptosis in sorafenib-resistant HCC cells. CONCLUSIONS: In this study, Torin2 and sorafenib showed synergistic cytostatic capacity in sorafenib-resistant HCC cells, via the suppression of mTORC2-AKT-BAD pathway. Our results suggest a novel strategy of drug combination for overcoming sorafenib resistance in HCC.

Lipopolysaccharide protects against acetaminophen-induced hepatotoxicity by reducing oxidative stress via the TNF-α/TNFR1 pathway.

Robust evidence suggested that gut-derived lipopolysaccharide (LPS) plays a significant role in various liver injury diseases; however, the role of gut-derived LPS in acetaminophen (APAP) overdose-induced acute liver injury remains unclear. The present study aimed to investigate the effect of gut-derived LPS on APAP-induced liver injury. Our results revealed that reduction of gut-derived LPS using multiple antibiotics could significantly exacerbate APAP-induced liver injury and increase mortality in mice. By contrast, pretreatment with exogenous LPS could reverse APAP-induced liver hepatotoxicity in mice and rats. We observed that TNF-α secretion in the liver was significantly increased after LPS pretreatment. In addition, depletion of TNF-α or TNFR1 inhibited the protective effects of LPS against APAP-induced hepatotoxicity, which indicated that the TNF-α/TNFR1 pathway was required to protect against APAP-induced liver injury. Mechanistically, LPS reduces oxidative stress by upregulating the expression of hepatic GSH, reducing MDA levels in liver tissues, and upregulating the expression of several antioxidant genes after APAP injection. However, the production of hepatic GSH was not enhanced in the liver tissues of rats lacking TNF-α or TNFR1 and MDA levels were not reduced after LPS and APAP co-treatment. The above results suggested LPS alleviated APAP-induced oxidative stress via the TNF-α/TNFR1 pathway.

Effect of biaxiality on chirality in chiral nematic liquid crystals.

Biaxiality and chirality are two of the most interesting topics in materials and biological science, particularly in liquid crystals. What is even more interesting is that these two properties are related. It was theoretically predicted that morphological chirality is impossible without morphological biaxiality in chiral nematic liquid crystals. We experimentally study the effect of biaxiality on chirality. We show that when achiral liquid crystal dimers with large molecular biaxiality are doped into chiral nematic liquid crystals, their helical pitch is significantly decreased. We have also observed an abnormal fingerprint texture of the chiral nematic liquid crystal doped dimers, which suggests morphological biaxiality. Our experimental results support the biaxial theory of chiral nematic liquid crystals.

Chemical bonding and Cu diffusion at the Cu/Ta2N interface: a DFT study.

Ta2N is an effective diffusion barrier material to prevent undesired Cu diffusion in ultra-large scale integration circuits. Previous theoretical work has reported the interesting result that at the Cu/Ta2N interface the Cu layer preferentially bonded with the Ta layer but not the N layer of Ta2N. However, this result was calculated from largely lattice mismatched interface models. To confirm this theoretical result and unravel the cause of strong Cu-Ta bonding at the Cu/Ta2N interface, in this study density functional theory calculations, on the basis of super-cell models, were performed to investigate the Cu(111)/Ta2N(001) interface. We firstly calculated interface cohesive energies and confirmed that the Cu layer preferentially bonded with the Ta layer of Ta2N. Then, electronic structure calculations revealed that the chemical bonding of the Cu-Ta bond at the Cu(111)/Ta2N(001) interface was primarily covalent in character, providing a proper explanation for the close integration of the Cu layer and Ta layer. Lastly, Cu diffusion investigations revealed that Ta2N was able to effectively prevent Cu diffusion. Furthermore, we found that the N layer of Ta2N played the critical role in preventing Cu diffusion.

Selective scattering polymer dispersed liquid crystal film for light enhancement of organic light emitting diode.

We developed a novel light enhancing film for an organic light emitting diode (OLED) based on polymer dispersed liquid crystal (PDLC). In the film, the liquid crystal droplets are unidirectionally aligned along the film normal direction and exhibit selective scattering. The film scatters light emitted only in directions with large incident angles but not light emitted in directions with small incident angles. When the light is scattered, it changes propagation direction and exits the OLED. The PDLC film reduces the total internal reflection and thus can significantly increase the light efficiency of the OLED.

Matched elastic constants for a perfect helical planar state and a fast switching time in chiral nematic liquid crystals.

Chiral nematic liquid crystals possess a self-assembled helical structure and exhibit unique selective reflection in visible and infrared light regions. Their optical properties can be electrically tuned. The tuning involves the unwinding and restoring of the helical structure. We carried out an experimental study on the mechanism of the restoration of the helical structure. We constructed chiral nematic liquid crystals with variable elastic constants by doping bent-dimers and studied their impact on the restoration. With matched twist and bend elastic constants, the helical structure can be restored dramatically fast from the field-induced homeotropic state. Furthermore, defects can be eliminated to produce a perfect planar state which exhibits high selective reflection.

Theoretical study on the surface stabilities, electronic structures and water adsorption behavior of the Ta3N5(110) surface.

A recent experiment revealed that the Ta3N5 semiconductor with orientation along the (110) surface exhibited improved photoelectrochemical activities, but the role of the (110) surface in the improved photoelectrochemical activity remains unclear. In this study, density functional theory calculations were performed to investigate the surface stabilities, surface electronic structures and water splitting behavior of the Ta3N5(110) surface with and without oxygen impurities. The results showed that, on the clean and oxygen impurity containing (110) surfaces, the energy barriers of water splitting were as low as 0.05 and 0.06 eV, respectively, suggesting that the Ta3N5(110) surface is a promising candidate for water splitting. The lower energy barriers of water splitting on the Ta3N5(110) surface may be ascribed to the easy migration of the H atom from the surface Ta atom to the nearby N atom. Furthermore, the surface energies and surface electronic structures revealed that the Ta3N5(110) surface contained less oxygen impurities, which is in accordance with the experimental observations.

Effects of oxygen impurities and nitrogen vacancies on the surface properties of the Ta3N5 photocatalyst: a DFT study.

Surface defects and impurities play important roles in the photocatalytic performance of semiconductors. In this study, DFT calculations are performed to investigate the effects of oxygen impurities and nitrogen vacancies on the surface stability and electronic structures of Ta3N5(100), (010) and (001) low-index surfaces. The results show that, for each surface, the oxygen impurities and nitrogen vacancies are beneficial and harmful, respectively, to the surface stability of Ta3N5. The oxygen impurities and nitrogen vacancies have mainly two effects on the surface electronic structures of Ta3N5. One is saturating surface states on the clean surface, and the other is inducing the downshift of conduction band minimum. In addition, the Ta3N5(100) surface with oxygen impurities is expected to have the strongest reduction ability in practice, providing useful guidance for further investigations of Ta3N5 in the photocatalytic hydrogen evolution.

Unraveling the mechanism of 720 nm sub-band-gap optical absorption of a Ta3N5 semiconductor photocatalyst: a hybrid-DFT calculation.

The Ta3N5 semiconductor photocatalyst possesses a 720 nm (about 1.72 eV) sub-band-gap optical absorption but the mechanism of this optical absorption is still controversial. In this study, the hybrid density functional theory calculations are performed to unravel the mechanism of 720 nm sub-band-gap optical absorption of Ta3N5. By studying the possible optical absorption initiated by the ON impurity and the VN defect, we find that the 720 nm sub-band-gap optical absorption of Ta3N5 may be ascribed to the electron transition from V(·)(N) to V(···)(N). In addition, we propose that the 720 nm sub-band-gap optical absorption can be used to qualitatively evaluate the photocatalytic water splitting ability of Ta3N5.

Overexpression of SIRT1 promotes metastasis through epithelial-mesenchymal transition in hepatocellular carcinoma.

BACKGROUND: SIRT1 is a member of the mammalian sirtuin family with the ability to deacetylate histone and nonhistone proteins. The correlation between SIRT1 expression and tumor metastasis in several types of cancer has aroused widespread concern. This study investigated SIRT1 expression and its prognostic value in hepatocellular carcinoma (HCC). The function of SIRT1 in hepatocarcinogenesis was further investigated in cell culture and mouse models. METHODS: Western blotting and immunohistochemistry were used to explore SIRT1 expression in HCC cell lines and primary HCC clinical specimens. The functions of SIRT1 in the migration and invasion in the HCC cell line were analyzed by infecting cells with adenovirus containing full-length SIRT1 or sh-RNA. The effect of SIRT1 on tumorigenicity in nude mice was also investigated. RESULTS: SIRT1 expression was significantly overexpressed in the tumor tissues and HCC cell lines. SIRT1 significantly promoted the ability of migration and invasion in HCC cells. In addition, experiments with a mouse model revealed that SIRT1 overexpression enhanced HCC tumor metastasis in vivo. Furthermore, we demonstrated that SIRT1 significantly enhanced the invasive and metastatic potential by inducing epithelial-mesenchymal transition in HCC cells. A clinicopathological analysis showed that SIRT1 expression was significantly correlated with tumor size, tumor number, and TNM staging. Kaplan-Meier survival curves revealed that positive SIRT1 expression was associated with poor prognosis in patients with HCC. CONCLUSIONS: Our data suggest that SIRT1 may play an important role in HCC progression and could be a potential molecular therapy target for HCC.

Moiré effect enables versatile design of topological defects in nematic liquid crystals.

Recent advances in surface-patterning techniques of liquid crystals have enabled the precise creation of topological defects, which promise a variety of emergent applications. However, the manipulation and application of these defects remain limited. Here, we harness the moiré effect to engineer topological defects in patterned nematic liquid crystal cells. Specifically, we combine simulation and experiment to examine a nematic cell confined between two substrates of periodic surface anchoring patterns; by rotating one surface against the other, we observe a rich variety of highly tunable, novel topological defects. These defects are shown to guide the three-dimensional self-assembly of colloids, which can conversely impact defects by preventing the self-annihilation of loop-defects through jamming. Finally, we demonstrate that certain nematic moiré cells can engender arbitrary shapes represented by defect regions. As such, the proposed simple twist method enables the design and tuning of mesoscopic structures in liquid crystals, facilitating applications including defect-directed self-assembly, material transport, micro-reactors, photonic devices, and anti-counterfeiting materials.

Self-compassion buffers the impact of learned helplessness on adverse mental health during COVID-19 lockdown.

BACKGROUND: Learned helplessness may be the underlying cause of poor mental health status among college students during the COVID-19 lockdown, and self-compassion as a positive psychological quality may influence the link between learned helplessness and mental health. METHODS: A sample of 869 Chinese college students (443 male and 426 female), with a mean age of 20.03 (SD = 1.68), completed the Learned Helplessness Scale (LHS), Self-Compassion Scale (SCS), and DASS-21. The moderating effect of self-compassion on the relationship between learned helplessness and anxiety, depression, and stress were calculated. RESULTS: The interaction term between learned helplessness and self-compassion has a significant coefficient on anxiety, depression, and stress, pointing out self-compassion as a moderator of the association between learned helplessness and adverse mental health. LIMITATIONS: In the absence of longitudinal data or experimental manipulations, cross-sectional methods cannot verify causal conclusions among the study variables. The analysing results are based only on self-reported data. DISCUSSION: The present study contributes to a deeper understanding of how learned helplessness and self-compassion during COVID-19 contribute to adverse mental health. The findings suggest that adverse mental health during lockdown is significantly associated with learned helplessness and that self-compassion can buffer this effect, contributing to future psychotherapy and clinical research. Future studies should examine the relationship through a longitudinal design to sort out whether self-compassion is a protective factor against learned helplessness or a moderator of the effects of learned helplessness on mental health.

Influence of surgical margin width on survival rate after resection of intrahepatic cholangiocarcinoma: a systematic review and meta-analysis.

OBJECTIVES: Hepatectomy is the best treatment for patients with intrahepatic cholangiocarcinoma (ICC) at present, but there has been controversy about the width of surgical margins. In this study, we systematically investigated the effects of different surgical margin widths on the prognosis of patients with ICC undergoing hepatectomy. DESIGN: Systematic review and meta-analysis. DATA SOURCES: PubMed, Embase and Web of Science databases were systematically searched from inception to June 2022. ELIGIBILITY CRITERIA: Cohort studies reported in English with patients who underwent negative marginal (R0) resection were included. The effects of surgical margin width on overall survival (OS), disease-free survival (DFS) and recurrence-free survival (RFS) in patients with ICC were assessed. DATA EXTRACTION AND SYNTHESIS: Two investigators independently conducted literature screening and data extraction. Risk of bias was assessed using funnel plots and quality was assessed by the Newcastle-Ottawa Scale. Forest plots of HRs and their 95% CIs for outcome indicators were plotted. Heterogeneity was assessed and determined quantitatively using I2, and the stability of the study results was evaluated using sensitivity analysis. Analyses were performed using Stata software. RESULTS: Nine studies were included. With the wide margin group (≥10 mm) as the control, pooled HR of OS in the narrow margin group (<10 mm) was 1.54 (95% CI 1.34 to 1.77). HRs of OS in three subgroups where the margin was less than 5 mm ranged from 5 mm to 9 mm, or was less than 10 mm in length were 1.88 (1.45 to 2.42), 1.33 (1.03 to 1.72) and 1.49 (1.20 to 1.84), respectively. Pooled HR of DFS in the narrow margin group (<10 mm) was 1.51 (1.14 to 2.00). Pooled HR of RFS in the narrow margin group (<10 mm) was 1.35 (1.19 to 1.54). HRs of RFS in three subgroups where the margin was less than 5 mm ranged from 5 mm to 9 mm, or was less than 10 mm in length were 1.38 (1.07 to 1.78), 1.39 (1.11 to 1.74) and 1.30 (1.06 to 1.60), respectively. Neither lymph node lesions (HR 1.44, 95% CI 1.22 to 1.70) nor lymph node invasion (2.14, 1.39 to 3.28) was favourable for postoperative OS in patients with ICC. Lymph node metastasis (1.31, 1.09 to 1.57) was unfavourable for RFS in patients with ICC. CONCLUSION: Patients with ICC who underwent curative hepatectomy with a negative margin ≥10 mm may have a long-term survival advantage, but lymph node dissection also needs to be considered. In addition, tumour-related pathological features need to be explored to see if they affect the surgical outcome of R0 margins.