Spatial proteomic landscape of primary and relapsed hepatocellular carcinoma reveals immune escape characteristics in early relapse.

BACKGROUND AIMS: Surgical resection serves as the principal curative strategy for hepatocellular carcinoma (HCC), yet the incidence of postoperative recurrence remains alarmingly high. However, the spatial molecular structural alterations contributing to postoperative recurrence in HCC are still poorly understood. APPROACH RESULTS: We employed imaging mass cytometry to profile the in-situ expression of 33 proteins within 358,729 single cells of 92 clinically annotated surgical specimens from 46 patients who were treated with surgical resections for primary and relapsed tumors. We revealed the recurrence progression of HCC was governed by the dynamic spatial distribution and functional interplay of diverse cell types across adjacent normal, tumor margin, and intratumor regions. Our exhaustive analyses revealed an aggressive, immunosuppression-related spatial ecosystem in relapsed HCC. Additionally, we illustrated the prominent implications of the TME of tumor margins in association with relapse HCC. Moreover, we identified a novel subpopulation of dendritic cells (PDL1+CD103+ DCs) enriched in the peritumoral area that correlated with early postoperative recurrence, which was further validated in an external cohort. Through the analysis of scRNA-seq data, we found the interaction of PDL1+CD103+ DCs with regulatory T cells and exhausted T cells enhanced immunosuppression and immune escape via multiple ligand-receptor pathways. CONCLUSIONS: We comprehensively depicted the spatial landscape of single-cell dynamics and multicellular architecture within primary and relapsed HCC. Our findings highlight spatial organization is a prominent determinant of HCC recurrence and provide a valuable insight into the immune evasion mechanisms driving recurrence.

Left Atrial Dissection Induced by Coronary Sinus Catheter-Related Injury.

We present a unique case of left atrial (LA) dissection in a 46-year-old man following aortic dissection surgery. The LA dissection was attributed to coronary sinus catheter-related injury. This report highlights the importance of recognizing this rare complication and the crucial role of transesophageal echocardiography in its diagnosis. We discuss the pathogenesis, diagnostic criteria, and management strategies for LA dissection.

Coupling Z-Scheme g-C3N4/rGO/MoS2 Ternary Heterojunction as an Efficient Visible Light Photocatalyst for Hydrogen Evolution and RhB Degradation.

Coupling heterostructures to synergistically improve the light adsorption and promote the charge carrier separation has been regarded as an operative approach to advance the photocatalytic performances. However, it is still challenging to construct heterostructures with appropriate optical properties and interfacial energy structures at the same time. In this work, a Z-scheme g-C3N4/rGO/MoS2 ternary composite photocatalyst is successfully synthesized via an effective hydrothermal method. The as-synthesized g-C3N4/rGO/MoS2 composite photocatalyst exhibited significant improvement for visible light absorption and boosted the separation efficiency of photoinduced electron-hole pairs. The g-C3N4/rGO/MoS2 system exhibited optimum visible-light-induced photocatalytic activity in hydrogen (H2) from water splitting and degrading pollutant rhodamin B (RhB), which is 22 times and 5 times higher than that of pure g-C3N4, respectively. The excellent photocatalytic activities are attributed to the synergetic effects of coupling rGO, g-C3N4, and MoS2 ternary structures to the composite photocatalyst. These combinations of intimate two-dimensional nanoconjugations can effectively inhibit charge recombination and accelerate charge transfer kinetics, forming a Z-scheme-assisted photocatalytic mechanism, thereby exhibiting superior photocatalytic activity.

Vimentin and tumor-stroma ratio for neoadjuvant chemoradiotherapy response prediction in locally advanced rectal cancer.

Vimentin expression in tumor tissues and the tumor-stroma ratio (TSR) have been demonstrated as strong prognostic factors for cancer patients, but whether they are predictive markers of neoadjuvant chemoradiotherapy (nCRT) outcome in locally advanced rectal cancer (LARC) patients is poorly understood. This study aimed to explore the predictive significance of vimentin and TSR combined for nCRT response in LARC patients. Imaging mass cytometry (IMC) was performed to determine the association of vimentin and TSR with nCRT response in six LARC patients [three achieved pathological complete response (pCR), three did not]. Immunohistochemistry (IHC) for vimentin and TSR on biopsy tissues before nCRT and logistic regression analysis were performed to further evaluate their predictive value for treatment responses in a larger patient cohort. A trend of decreased vimentin expression and increased TSR in the pCR group was revealed by IMC. In the validation group, vimentin [odds ratio (OR) 0.260, 95% confidence interval (CI) 0.102-0.602, p = 0.002] and TSR (OR 4.971, 95% CI 1.933-15.431, p = 0.002) were associated with pCR by univariate analysis. Patients in the vimentin-low/TSR-low or vimentin-high/TSR-high (OR 5.211, 95% CI 1.248-35.582, p = 0.042) and vimentin-low/TSR-high groups (OR 11.846, 95% CI 3.197-77.079, p = 0.001) had significantly higher odds of pCR. By multivariate analysis, only the combination of vimentin and TSR was an independent predictor for nCRT response (OR 9.324, 95% CI 2.290-63.623, p = 0.006). Our study suggested that the combined assessment of vimentin and TSR can provide additive significance and may be a promising indicator of nCRT response in LARC patients.

Contribution of Oxidative Stress Induced by Sonodynamic Therapy to the Calcium Homeostasis Imbalance Enhances Macrophage Infiltration in Glioma Cells.

BACKGROUND: To better understand the Ca2+ overload mechanism of SDT killing gliomas, we examined the hypothesis that the early application of the mechanosensitive Ca2+ channel Piezo1 antagonist (GsMTx4) could have a better anti-tumor effect. METHODS: The in vitro effect of low-energy SDT combined with GsMTx4 or agonist Yoda 1 on both the ROS-induced distribution of Ca2+ as well as on the opening of Piezo1 and the dissociation and polymerization of the Ca2+ lipid complex were assessed. The same groups were also studied to determine their effects on both tumor-bearing BALB/c-nude and C57BL/6 intracranial tumors, and their effects on the tumor-infiltrating macrophages were studied as well. RESULTS: It was determined that ultrasound-activated Piezo1 contributes to the course of intracellular Ca2+ overload, which mediates macrophages (M1 and M2) infiltrating under the oxidative stress caused by SDT. Moreover, we explored the effects of SDT based on the dissociation of the Ca2+ lipid complex by inhibiting the expression of fatty acid binding protein 4 (FABP4). The Piezo1 channel was blocked early and combined with SDT treatment, recruited macrophages in the orthotopic transplantation glioma model. CONCLUSIONS: SDT regulates intracellular Ca2+ signals by upregulating Piezo1 leading to the inhibition of the energy supply from lipid and recruitment of macrophages. Therefore, intervening with the function of the Ca2+ channel on the glioma cell membrane in advance is likely to be the key factor to obtain a better effect combined with SDT treatment.

Bioinformatics analysis of differentially expressed proteins in alcoholic fatty liver disease treated with recombinant human cytoglobin.

Cytoglobin (Cygb) is a globin molecule that is ubiquitously expressed in all tissues and has a protective role under oxidative stress. It has also been demonstrated to be effective in the treatment of alcoholic fatty liver disease (AFLD). In order to study the molecular mechanisms underlying its beneficial effects for the treatment of alcoholic liver, two­dimensional electrophoresis and mass spectrometric analysis were performed on serum and liver tissues from an in vivo rat model of AFLD. A total of 26 differentially expressed proteins were identified in the serum and 20 differentially expressed proteins were identified in liver specimens. Using online bioinformatics tools, it was indicated that these differentially expressed proteins were primarily associated with pathways including binding and uptake of ligands by scavenger receptors, response to corticosteroid, plasma lipoprotein remodeling, regulation of complement cascade, hydrogen peroxide catabolic process, as well as response to nutrient and monosaccharide. The present results suggested that recombinant human Cygb exerts its role in the treatment of AFLD primarily through affecting nutrient metabolism, monocarboxylic acid biosynthesis, regulation of glutathione expression, plasma lipoprotein remodeling and removal of metabolic waste from the blood.

Risk factors of delayed awakening after aortic arch surgery under deep hypothermic circulatory arrest with selective antegrade cerebral perfusion.

BACKGROUND: To determine the risk factors of delayed awakening following aortic arch surgery under deep hypothermic circulatory arrest (DHCA) in combination with selective antegrade cerebral perfusion (SACP). METHODS: We retrospectively analyzed the clinical data of all patients who underwent aortic arch surgery under DHCA + SACP between September 2015 and September 2017 in our hospital. Delayed awakening was defined as recovery of consciousness later than 24 hours after the surgery. Risk factors of delayed awakening were evaluated using multivariate logistic regression analysis. RESULTS: A total of 168 subjects were included. In-hospital mortality of the overall sample was 19.05% (n=32). Delayed awakening occurred in 76 (45.23%) subjects. Subjects with delayed awakening had older age, hypertension, higher rate of emergency surgery and blood transfusion, and longer cardiopulmonary bypass (CPB) time and myocardial blocking time. Multivariate regression analysis showed emergency surgery (P=0.005) and CPB time >240 min (P<0.001) as risk factors for delayed awakening, even after adjusting potential confounders, including age, hypertension, aortic cross-clamp time and blood transfusion. CONCLUSIONS: In patients undergoing aortic arch surgery under DHCA + SACP, emergency surgery and CPB time >240 min are risk factors for delayed awakening.

[Eukaryotic expression, purification of human IgECepsilon2-4 protein and its target fishing].

Objective To construct eukaryotic expression vectors of human IgE heavy chain 2-4 region (IgECepsilon2-4) and purify the recombinant protein, and then capture its interacted proteins by surface plasmon resonance (SPR). Methods Three recombinant eukaryotic expression vectors of IgECepsilon2-4 containing different signal peptides were constructed and transiently transfected into HEK293FT suspension cells separately. The recombinant plasmid with the highest-level expression was selected to express the recombinant protein in a huge amount, and then the recombinant protein was purified by Ni-NTA affinity chromatography. The interaction between high-affinity IgE receptor (FcepsilonR I) of KU812 cell surface and IgECepsilon2-4 was identified by immunofluorescence cytochemistry. The unknown proteins that specifically interacted with IgECepsilon2-4 were captured from human serum by SPR technique. Results The recombinant plasmid containing the signal peptide III showed the highest expression (6.2 mg/L). Highly purified recombinant protein IgECepsilon2-4 was obtained by affinity purification. Immunofluorescence cytochemistry showed that the recombinant protein IgECepsilon2-4 could be combined with the surface receptor of KU812 cells. Thirty-nine kinds of proteins which were likely to interact with IgECepsilon2-4 were captured from human serum by SPR. Conclusion We obtained the purified recombinant protein IgECepsilon2-4 that could be combined with KU812 cell surface receptor. Target fishing experiment revealed that the recombinant protein IgECepsilon2-4 was likely to interact with 39 kinds of proteins in human serum.

[Effect of alendronate on interleukin-1beta induced chondrocytes of rat in vitro].

OBJECTIVE: To investigate the feasibility of alendronate (ALN) in treating osteoarthritis (OA) by observing the effects of ALN on interleukin 1beta (IL-1beta) induced chondrocytes of rat in vitro. METHODS: The chondrocytes of knee articular surface from 15 SD rats (1-month-old, female or male, weighing 100-150 g) were cultured. The chondrocytes were observed by inverted phase contrast microscope and identified by toluidine blue staining and HE staining. The third passage chondrocytes were divided into 3 groups: the chondrocytes were cultured with DMEM for 5 days in group A, with 10 ng/mL IL-1beta for 2 days and with DMEM for 3 days in group B, and with 10 ng/mL IL-1beta for 2 days and with 1 x 10(6) mol/L ALN for 3 days in group C. Immunocytochemistry and real-time PCR were performed to determine the expression levels of collagen type II (Col II), matrix metalloproteinase 13 (MMP-13), and p-catenin. RESULTS: Toluidine blue staining proved that the cultured cells were chondrocytes. The integrated absorbency (IA) value of Col II in group C (10.290 7 +/- 0.499 2) was lower than that in group A (15.377 0 +/- 0.571 8) and higher than that in group B (5.463 2 +/- 0.450 4), showing significant differences (P < 0.05). The IA value of MMP-13 in group C (3.068 6 +/- 0.205 6) was significantly lower than that in group B (6.998 1 +/- 0.329 7, P < 0.05), but there was no significant difference when compared with group A (2.777 5 +/- 0.199 6, P > 0.05). The IA value of beta-catenin in group C (6.611 7 +/- 0.381 8) was lower than that in group B (11.799 9 +/- 0.348 7) and higher than that in group A (4.390 3 +/- 0.551 9), showing significant differences (P < 0.05). The mRNA expression of Col II in group C was significantly higher than those in groups A and B (P < 0.05), the mRNA expression of MMP-13 in group C was significantly lower than that in group B (P < 0.05) but there was no significant difference when compared with group A (P > 0.05). The mRNA expression of j-catenin in group C was significantly lower than that in group B (P < 0.05) and higher than that in group A (P < 0.05). CONCLUSION: ALN can protect rat chondrocyte from OA induced by IL-1beta in vitro possibly by upregulating Col II and inhibiting the expression of MMP-13 and beta-catenin in the chondrocytes.

[In vitro effect of alendronate on chondrocytes and articular cartilage and subchondral bone in rabbit anterior cruciate ligament transection model].

OBJECTIVE: To examine the effects of alendronate (ALN) on IL-1beta-stimulated chondrocyte of rabbit in vitro and on cartilage and subchondral bone in rabbit osteoarthritis (OA) induced by anterior cruciate ligament transection (ACLT). METHODS: The chondrocytes from articular surface of healthy 3-month-old Japanese White rabbits were obtained by the method of enzyme digestion and cultured in vitro. The third generation chondrocytes were assigned into three groups: the chondrocytes were cultured in DMEM medium with 10 ng/mL IL-1beta for 2 days, subsequently with (ALN group, group A1) or without (IL-1beta group, group B1) 1 x 10(-6) mol/L ALN for 3 days; the chondrocytes in vacant group (group C1) were cultured in DMEM medium for 5 days. The expression of Col II and MMP-13 were analyzed by immunocytochemical staining observation and real time RT-PCR test. Another twenty-four 3-month-old male Japanese White rabbits were randomized into three groups (n = 8 per group). The OA model was made by ACLT in ACLT+ALN group (group A2) and ACLT group (group B2); the joint cave was sutured after exposure of ACL in sham group (group C2). After 4 days, the rabbits of group A2 received the subcutaneous injection of ALN at a dosage of 10 microg/(kg x d) for 8 weeks. Rabbits of group B2 and C2 received equal normal saline treatment. After 8 weeks, the rabbits were executed. The macro-pathologic changes of right knee joints were observed, so were the histological changes of femoral condyles. Expression levels of Col II and MMP-13 were detected by immunohistochemical staining. The bone histomorphometry analysis was applied to subchondral bone of proximal tibia. RESULTS: In vitro, the Col II immunocytochemical staining showed intensely positive staining in group C1, and the intensity of staining was slightly decreased in group A1, but the intensity of Col II immunocytochemical staining was extremely lower in the group B1. The integrated absorbance (IA) value for Col II in group A1 was significantly higher than that of group B1 (P < 0.05), but there was no significant difference between group A1 and group C1 (P > 0.05). Immunocytochemical detection of MMP-13 showed intense staining in group B1, and the intensity of staining was slightly decreased in group A1, but no MMP-13 expression was detected in the group C1. The IA value for MMP-13 in group A1 was significantly lower than that of group B1 (P < 0.05), but significantly higher than that of group C1 (P <0.05). The real time RT-PCR analysis showed significantly higher mRNA levels of Col II in group A1 than in group B1 (P < 0.05), but there was no significant difference between group A1 and group C1 (P > 0.05). The MMP-13 mRNA level of the chondrocytes in group A1 was significantly lower than that of group B1 (P < 0.05), but significantly higher than that of group C1 (P < 0.05). In vivo, the gross appearance of surface of knee joint showed that there was no ulcer in group C2, and there was some ulcers in group A2, but many and all layers ulcers in group B2. Mankin score of group A2 was significantly lower than that of group B2 (P < 0.05), but significantly higher than that of group C2 (P < 0.05). Immunohistochemical staining showed that Col II in articular cartilage was intensely staining in group C2, the intensity of staining was slightly decreased in group A2, and the intensity of Col II immunohistochemical staining was extremely low in group B2, but there was no significant difference between group A2 and group C2 (P > 0.05). The immunohistochemical staining for MMP-13 significantly increased in group B2, the intensity of staining was slightly decreased in group A2, and no MMP-13 expression was detectable in the group C2. The IA value for MMP-13 in group A2 was significantly lower than that of group B2 (P < 0.05), but significant higher than that of group C2 (P < 0.05). Bone histomorphometry showed that in group B2 percent trabecular area and trabecular thickness markedly decreased compared with those in group A2 and group C2 (P < 0.05), but there was no significant difference between group A2 and group C2 (P > 0.05). The trabecular separation, percent labeled perimeter and bone formation rate were significantly elevated in group B2 compared with those in group A1 and group C2 (P < 0.05), but there was no significant difference between group A2 and group C2 (P > 0.05). No significant difference was evident on trabecular number and mineral apposition rate values among groups A2, B2, and C2 (P > 0.05). CONCLUSION: For rabbits OA induced by ACLT, the subcutaneous injections of alendronate can inhibit cartilage degradation, prevent bone loss, and improve microarchitecture of subchondral bone. ALN can partially protect chondrocytes by inhibiting the expression of MMP-13 both in vitro and in vivo.

[Pulmonary alveolar proteinosis treated with whole-lung lavage utilizing extracorporeal membrane oxygenation: a case report and review].

OBJECTIVE: To improve the treatment of severe hypoxaemia in patients with pulmonary alveolar proteinosis (PAP). METHODS: The clinical data of a patient with pathologically proven PAP treated with whole-lung lavage utilizing extracorporeal membrane oxygenation (ECMO) were described and the literature was reviewed. RESULTS: This 57-year-old man was admitted because of cough and progressive dyspnea for 12 months. His PaO(2) was 46 mm Hg (1 mm Hg = 0.133 kPa) and saturation of pulse oximeter (SpO(2)) was from 85% to 88% with oxygen 5 L/min by nasal cannula. His chest CT, bronchoscopy with bronchoalveolar lavage and transbronchial biopsies were consistent with PAP. Whole-lung lavage was performed in the operation room under general anesthesia. A double-lumen tube was intubated in order to selectively ventilate and lavage a single lung independently. During mechanical ventilation for both lungs, the SpO(2) was from 80% to 90%, but when a single right lung ventilation was tried, the SpO(2) (from 68% to 80%) dropped significantly. To ensure adequate oxygen supply during lavage, a veno-arterial ECMO was set up by inserting catheters percutaneously into the right femoral artery and right femoral vein respectively. Then the SpO(2) improved, from 89% to 97% during single right lung ventilation. The left lung was lavaged with a total of 20.8 L of normal saline. The SpO(2) ranged from 80% to 94% during the lavage. After the lavage, the patient no longer experienced shortness of breath. Then 28 days later the right lung was lavaged without the aid of ECMO. A month after the second lavage, his chest CT showed marked improvement in infiltrates of both lungs. CONCLUSION: When a patient with PAP has refractory hypoxemia prior to the lavage procedure, ECMO should be considered in order to avoid severe hypoxaemia with fatal consequences during lavage.