[Pathological features related to onco-immunity and their clinical significance of pancreatic ductal adenocarcinoma].

Objective: To investigate the tumor immunity-related pathologic features and clinical significance in pancreatic ductal adenocarcinoma (PDAC). Methods: All pathologic materials and clinical information of 192 PDAC patients from the Cancer Hospital of the University of Chinese Academy of Sciences from January 2010 to December 2020 were collected. The onco-immune microenvironment associated morphologic features were evaluated, and MHC-Ⅰ, PD-L1, CD3, and CD8 expression were detected by immunohistochemistry (IHC). Then the correlation between the factors and their influence on prognosis was analyzed. Results: There were 163 cases of non-specific adenocarcinoma (163/192, 84.90%), 18 cases of adeno-squamous carcinoma (18/192, 9.37%), and 11 cases of other rare subtypes (11/192, 5.73%). Perineural invasion was observed in 110 cases (110/192, 57.29%) and vascular invasion in 86 cases (86/192, 44.79%). There were 84 cases (84/182, 46.15%) with severe chronic inflammation. Tumor infiltrating immune cell numbers (TII-N) were increased in 52 cases (52/192, 27.08%). Lymphocytes and plasma cells were the main infiltrating immune cells in 60 cases (60/192, 31.25%), whereas in 34 cases (34/192, 17.71%) the tumors were mainly infiltrated by granulocytes, and 98 cases (98/192, 51.04%) showed mixed infiltration. CD3+T cells were deficient in 124 cases (124/192, 66.31%). CD8+T cells were deficient in 152 cases (152/192, 79.58%). MHC-Ⅰ expression was down-regulated in 156 cases (156/192, 81.25%), and PD-L1 was positive (CPS≥1) in 46 cases (46/192, 23.96%). Statistical analysis showed that TII-N was negatively correlated with vascular invasion (P=0.035), perineural invasion (P=0.002), stage (P=0.004) and long-term alcohol consumption (P=0.039). The type of immune cells correlated positively with chronic pancreatic inflammation (P=0.002), and negatively with tumor differentiation (P=0.024). CD8+T cells were positively correlated with CD3+T cells (P=0.032), MHC-Ⅰ expression (P<0.001) and PD-L1 expression (P=0.001), and negatively correlated with long-term smoking (P=0.016). Univariate analysis showed that histological nonspecific type (P=0.013) and TII-N (P<0.001) were the factors for good prognosis. Vascular invasion (P=0.032), perineural invasion (P=0.001), high stage (P=0.003) and long-term alcohol consumption (P=0.004) were adverse prognostic factors. COX multivariate risk analysis found that TII-N was an independent favorable factor for PDAC, while perineural invasion was an independent adverse risk factor. Conclusions: TII-N is an independent superior prognostic factor for PDAC, and significantly correlated with many factors; chronic alcohol consumption and smoking may inhibit onco-immunity in PDAC patients.

Nitrogen-plasma doping of carbon film for a high-quality layered Si/C composite anode.

The effect of the chemical component and microstructure, not to mention their facile modification, of the coating/wrapping carbon layer on the electrochemical performance of the Si/C composite anode in lithium ion batteries (LIBs) hasn't been actively explored although Si/C has been recognized as one of the most promising route for the high energy density LIBs. Herein we propose a novel nitrogen-plasma doping route to modify the top carbon film in an elaborately constructed layered Si/C composite anode. The electrochemical performance, e.g., the initial coulombic efficiency (CE), cycle stability and specific capacity of the composite anode is drastically improved by this plasma processing due to the increased kinetics of lithium ions. By means of the appropriate adjustment of the N doping ratio and N chemical configuration in the carbon layer through a N2/H2 plasma processing, the lithium diffusion rate in the composite anode was memorably increased as the pseudocapacitance effects promoted. The optimized Si/C composite exhibits a high capacity of 1120.7 mA h g-1 and an initial CE of 80.8% at the current of 2 A g-1 after a long cycle of 1500, increasing by ~40% of specific capacity and ~29% of the initial CE.

All in one plasma process: From the preparation of S-C composite cathode to alleviation of polysulfide shuttle in Li-S batteries.

Tremendous efforts have been made to improve the electrochemical performance of the lithium-sulfur batteries. However, challenges remain in achieving fast electronic and ionic transport while accommodate the significant cathode volumetric change. On the other hand, the severe capacity decay mainly attributed to polysulfide shuttle also hampers the practical applications. Here, we report a simple, low-cost, and eco-friendly method for the one-step preparation of a binder-free S-C composite cathode by plasma dissociation of CS2 containing gases at room-temperature. The key issue of polysulfide shuttle effect in Li-S batteries is also effectively resolved just by the introduction of N2 into the precursor gases. The electrode exhibits a high reversible capacity of ~600 mAh/g of the total hybrid of S + C at 100 mA/g after 100 cycles with an excellent initial coulombic efficiency of nearly 100%. The cells also demonstrate along cycle life and an extremely high capacity of ~306 mAh/g even after 300 cycles at 1 A/g with a high coulombic efficiency of about 100%. The proposed method will open the way for the plasma applications in facile preparation of Li-S batteries and the improvement of its electrochemical performance.

[Effects of hydrogen on the lung damage of mice at early stage of severe burn].

Objective: To investigate the effects of hydrogen on the lung damage of mice at early stage of severe burn. Methods: One hundred and sixty ICR mice were divided into sham injury, hydrogen, pure burn, and burn+ hydrogen groups according to the random number table, with 40 mice in each group. Mice in pure burn group and burn+ hydrogen group were inflicted with 40% total body surface area full-thickness scald (hereafter referred to as burn) on the back, while mice in sham injury group and hydrogen group were sham injured. Mice in hydrogen group and burn+ hydrogen group inhaled 2% hydrogen for 1 h at post injury hour (PIH) 1 and 6, respectively, while mice in sham injury group and pure burn group inhaled air for 1 h. At PIH 24, lung tissue of six mice in each group was harvested, and then pathological changes of lung tissue were observed by HE staining and the lung tissue injury pathological score was calculated. Inferior vena cava blood and lung tissue of other eight mice in each group were obtained, and then content of high mobility group box 1 (HMGB1) and interleukin-6 (IL-6) in serum and lung tissue was determined by enzyme-linked immunosorbent assay. Activity of superoxide dismutase (SOD) in serum and lung tissue was detected by spectrophotometry. After arterial blood of other six mice in each group was collected for detection of arterial partial pressure of oxygen (PaO(2)), the wet and dry weight of lung tissue were weighted to calculate lung wet to dry weight ratio. The survival rates of the other twenty mice in each group during post injury days 7 were calculated. Data were processed with one-way analysis of variance, LSD test and log-rank test. Results: (1) At PIH 24, lung tissue of mice in sham injury group and hydrogen group showed no abnormality. Mice in pure burn group were with pulmonary interstitial edema, serious rupture of alveolar capillary wall, and infiltration of a large number of inflammatory cells. Mice in burn+ hydrogen group were with mild pulmonary interstitial edema, alveolar capillary congestion accompanied by slight rupture and bleeding, and the number of infiltration of inflammatory cells was smaller than that in pure burn group. The lung tissue injury pathological scores of mice in sham injury group, hydrogen group, pure burn group, and burn+ hydrogen group were (0.7±0.5), (0.8±0.5), (6.1±1.0), and (2.8±0.8) points, respectively. The lung tissue injury pathological score of mice in pure burn group was significantly higher than that in sham injury group (P<0.001). The lung tissue injury pathological score of mice in burn+ hydrogen group was significantly lower than that in pure burn group (P<0.001). (2) At PIH 24, the content of HMGB1 and IL-6 in serum and lung tissue of mice in hydrogen group was close to that in sham injury group (with P values above 0.05). The content of HMGB1 and IL-6 in serum and lung tissue of mice in pure burn group was significantly higher than that in sham injury group (with P values below 0.001). The content of HMGB1 and IL-6 in serum and lung tissue of mice in burn+ hydrogen group was significantly lower than that in pure burn group (with P values below 0.001). (3) At PIH 24, the activity of SOD in serum and lung tissue of mice in hydrogen group was close to that in sham injury group (with P values above 0.05). The activity of SOD in serum and lung tissue of mice in pure burn group was significantly lower than that in sham injury group (with P values below 0.001). The activity of SOD in serum and lung tissue of mice in burn+ hydrogen group was significantly higher than that in pure burn group (with P values below 0.001). (4) At PIH 24, there was no statistically significant difference in PaO(2) among the mice in four groups (F=0.04, P>0.05). (5) At PIH 24, the ratios of lung wet to dry weight of mice in sham injury, hydrogen, pure burn, and burn+ hydrogen groups were 3.52±0.22, 3.61±0.24, 7.24±0.32, and 5.21±0.41, respectively. The ratio of lung wet to dry weight of mice in pure burn group was significantly higher than that in sham injury group (P<0.001). The ratio of lung wet to dry weight of mice in burn+ hydrogen group was significantly lower than that in pure burn group (P<0.001). (6) The survival rates of mice in sham injury group and hydrogen group during post injury days 7 were 100%. Compared with those in sham injury group, survival rates of mice in pure burn group from post injury days 3 to 7 were significantly decreased (with P values below 0.05). Compared with those in pure burn group, survival rates of mice in burn+ hydrogen group from post injury days 5 to 7 were significantly increased (with P values below 0.05). Conclusions: Hydrogen can significantly alleviate the infiltration of inflammatory cells and improve the pathological lesions of lung tissue of mice with severe burn. It has the effects of reducing inflammatory reaction and inhibiting oxidative stress, further showing the protective effect on the lung of burn mice.

[Influence of HIF- 2α on the expression of GATA- 1 in bone marrow CD71(+) cell of high altitude polycythemia rat model].

OBJECTIVE: To explore the influence of hypoxia-inducible factor-2 αlpha (HIF-2α) on the expression of erythroid-specific transcription factor GATA-1 in bone marrow CD71(+) cells of rat model with high altitude polycythemia (HAPC). METHODS: A total of 48 male SD rats were selected and randomly divided into normal control group and HAPC group. HAPC model was established at an altitude of 4 300 meters in the natural environment and verified by bone marrow cell classification and counting, hematologic parameters and serum EPO detection. Bone marrow CD71 (+) cells were separated by a combination of methods with density gradient centrifugation and magnetic activated cell sorting. The changes of expression level of HIF-2α, GATA-1 mRNA and proteins were detected by Q-PCR and Western blot. CD71 (+) cells were cultured under hypoxia condition and transfected with selected optimal HIF- 2α shRNAi3 for 96 h. And the expression level of HIF-2α and GATA-1 mRNA and proteins were detected by Q- PCR and Western blot. RESULTS: The results of bone marrow cell counts, the hematologic parameters and the serum EPO content showed that the HAPC rat model was successfully established. The expression of HIF-2α and GATA-1 mRNA and protein in bone marrow CD71(+) cells of HAPC group was higher than that in control group (P<0.05). And HIF-2α and GATA-1 of HAPC group were positively correlated at the expression levels of mRNA and protein, respectively (r=0.923, P<0.01; r=0.838, P<0.01). However, the expression of HIF-2α and GATA-1 mRNA and protein in HAPC group was significantly lower than that in control groups after interfered by HIF-2α shRNAi3 for 96 h (P<0.05). CONCLUSION: The effect of HIF-2α on GATA-1 expression may be correlated with the pathogenesis of HAPC.

Monitoring of atherosclerosis evolution by detection of inflammatory states of aortae in a rabbit model using 18F-FDG -PET/CT.

AIM: In vivo dynamic evaluation of atherosclerosis could be clinically significant in the prevention of cardiovascular events. We aimed to monitor Fluorine-18 fluorodeoxyglucose (18F-FDG) uptake in different stages of atherosclerosis, and investigate the feasibility of detecting vulnerable plaques using positron emission tomography/computed tomography (PET/CT) angiography. METHODS: Twenty-two male NZW rabbits were divided into two groups: atherosclerosis group (group A, N.=11) and atherosclerosis and statin group (group S, N.=11). The rabbits underwent two pharmacological triggerings to induce thrombus at the 18th week. In vivo PET/CT scans were performed on four time points: before cholesterol diet (baseline, N.=6), at 8th week (the middle-of-feeding, N.=4), at 18th week (the end-of-feeding, N.=22) and after triggering (post-triggering, N.=15). 18F-FDG uptake by the aorta was expressed as maximal standardized uptake value (SUVmax) and mean SUV (SUVmean). SUVs were measured on serial 7.5 mm arterial segments. RESULTS: SUVmean and SUVmax were 0.449±0.108 and 0.550±0.132 at baseline, 0.694±0.117 and 0.754±0.129 at the middle-of-feeding, 0.788±0.121 and 0.861±0.139 in group A, and 0.651±0.194 and 0.736±0.243 in group S at the end-of feeding before triggering. SUVmean and SUVmax were 1.128±0.420 and 1.302±0.489 in thrombosis group, 0.774±0.159 and 0.859±0.191 in non-thrombosis group after triggering. Thrombus were identified in 10 of 22 rabbits (45.5%): 8 of 11 (72.3%) in group A, and 2 of 11 (18.2%) in group S (P<0.001). CONCLUSION: The inflammatory states of atherosclerosis and vulnerable plaque can be detected by quantitative analysis of 18F-FDG uptake. PET/CT may be used for predicting thrombosis events in patients with atherosclerotic disease.

Genome-wide analysis of DNA methylation variations caused by chronic glucolipotoxicity in beta-cells.

There is a growing body of literature suggesting the role of interactions between genes and the environment in development of type 2 diabetes mellitus (T2DM). However, the interplay between environment and genetic in developing and progressing T2MD is not fully understood. To determine the effects of high-glucose-lipid on the status of DNA methylation in beta cells, and clarify the mechanism of glucolipotoxicity on beta-cell deterioration, the DNA methylation profile was detected in beta-cells cultured with high-glucose-lipid medium.We utilized a high throughput NimbleGen RN34 CpG Island & Promoter Microarray to investigate the DNA methylation profile in beta-cells cultured with high-glucose-lipid medium. To validate the results of microarray, the immunoprecipitation (MeDIP) PCR was used to test the methylation status of some selected genes. The mRNA and protein expression of insulin and Tcf7l2 in these cells were quantified by RT-PCR and western blot, respectively.We have identified a lot of loci which experienced aberrant DNA methylation in beta-cells cultured with high-glucose-lipid medium. The results of MeDIP PCR were consistency to the microarray. An opposite regulation in transcription and translation of Tcf7l2 gene was found. Furthermore, the insulin mRNA and protein expression in beta-cells also decreased after cultured with high-glucose-lipid medium compared with the control cells.We conclude that chronic glucolipotoxicity could induce aberrant DNA methylation of some genes and may affect these genes expression in beta-cells, which might contribute to beta-cell function failure in T2DM and be helpful to explain, at least partially, the mechanism of glucolipotoxicity on beta-cells deterioration.

Novel monoclonal antibodies to ESAT-6 and CFP-10 antigens for ELISA-based diagnosis of pleural tuberculosis.

OBJECTIVE: To elucidate the potential of monoclonal antibodies (mAbs) of culture filtrate protein 10 (CFP-10) and early secretory antigenic target 6 (ESAT-6) in tuberculosis (TB) diagnosis. DESIGN: We generated and characterised monoclonal and polyclonal antibodies against Mycobacterium tuberculosis-specific antigens ESAT-6 and CFP-10 by immunising BALB/c mice with an ESAT-6/CFP-10 fusion protein. Stable hybridoma cell lines were established and mAbs were specifically identified by immunoblotting and immunoprecipitation. The mouse mAbs were used to coat plates, and biotin-labelled polyclonal antibodies were used to detect the antigens. One hundred and seventy-three samples of sputum culture supernatants and pleural effusion aspirates have been tested. RESULTS: The ESAT-6 enzyme-linked immunosorbent assay (ELISA) detected the culture supernatants and pleural effusion specimens that were positive for M. tuberculosis, but failed to identify M. tuberculosis-positive specimens in the non-M. tuberculosis culture supernatants or control specimens. This yielded a sensitivity of 95.4% and a specificity of 100% for the ESAT-6-specific ELISA. The CFP-10 ELISA presented less satisfactory sensitivity and specificity, of respectively 81.6% and 92.2%. Results showed positive detection rates of ESAT-6 and CFP-10 of 86.8% (33/38) and 76.3% (29/38) for the diagnosis of tuberculous pleural effusion in patients bacteriologically negative for M. tuberculosis culture. CONCLUSION: The ESAT-6 and CFP-10 ELISAs incorporating mAbs generated in this study serve as potential tools in the laboratory diagnosis of TB.