Orbital- and millennial-scale Asian winter monsoon variability across the Pliocene-Pleistocene glacial intensification.

Intensification of northern hemisphere glaciation (iNHG), ~2.7 million years ago (Ma), led to establishment of the Pleistocene to present-day bipolar icehouse state. Here we document evolution of orbital- and millennial-scale Asian winter monsoon (AWM) variability across the iNHG using a palaeomagnetically dated centennial-resolution grain size record between 3.6 and 1.9 Ma from a previously undescribed loess-palaeosol/red clay section on the central Chinese Loess Plateau. We find that the late Pliocene-early Pleistocene AWM was characterized by combined 41-kyr and ~100-kyr cycles, in response to ice volume and atmospheric CO2 forcing. Northern hemisphere ice sheet expansion, which was accompanied by an atmospheric CO2 concentration decline, substantially increased glacial AWM intensity  and its orbitally oscillating amplitudes across the iNHG. Superposed on orbital variability, we find that millennial AWM intensity fluctuations persisted during both the warmer (higher-CO2) late Pliocene and colder (lower-CO2) early Pleistocene, in response to both external astronomical forcing and internal climate dynamics.

Extraction, purification, structural features, modifications, bioactivities, structure-activity relationships, and applications of polysaccharides from garlic: A review.

Garlic is a common vegetable and spice in people's daily diets, in which garlic polysaccharide (GP) is one of the most important active components with a variety of benefits, such as antioxidant, immune-enhancing, anti-inflammatory, liver-protective and bowel-regulating properties. >20 types of GPs, mainly crude polysaccharides, have been identified. However, the exact chemical composition of GPs or the mechanism underlying their pharmacological activity is still not fully understood. The extraction and purification methods of GPs are compared in this review while providing detailed information on their structural features, identification methods, major biological activities, mechanisms of actions, structural modifications, structure-activity relationships as well as potential applications. Finally, the limitations of GP research and future issues that need to be addressed are discussed in this review. GPs are widely recognized as substances with great potential in the pharmaceutical and food industries. Therefore, this review aims to provide a comprehensive summary of the latest research progresses in the field of GPs, together with scientific insights and a theoretical support for the development of GPs in research and industrialization.

Concurrent Asian monsoon strengthening and early modern human dispersal to East Asia during the last interglacial.

The relationship between initial Homo sapiens dispersal from Africa to East Asia and the orbitally paced evolution of the Asian summer monsoon (ASM)-currently the largest monsoon system-remains underexplored due to lack of coordinated synthesis of both Asian paleoanthropological and paleoclimatic data. Here, we investigate orbital-scale ASM dynamics during the last 280 thousand years (kyr) and their likely influences on early H. sapiens dispersal to East Asia, through a unique integration of i) new centennial-resolution ASM records from the Chinese Loess Plateau, ii) model-based East Asian hydroclimatic reconstructions, iii) paleoanthropological data compilations, and iv) global H. sapiens habitat suitability simulations. Our combined proxy- and model-based reconstructions suggest that ASM precipitation responded to a combination of Northern Hemisphere ice volume, greenhouse gas, and regional summer insolation forcing, with cooccurring primary orbital cycles of ~100-kyr, 41-kyr, and ~20-kyr. Between ~125 and 70 kyr ago, summer monsoon rains and temperatures increased in vast areas across Asia. This episode coincides with the earliest H. sapiens fossil occurrence at multiple localities in East Asia. Following the transcontinental increase in simulated habitat suitability, we suggest that ASM strengthening together with Southeast African climate deterioration may have promoted the initial H. sapiens dispersal from their African homeland to remote East Asia during the last interglacial.

Development of a system for detecting cardiac troponin I by background fluorescence quenching based on internal filtration effect.

The present study sought to develop a cardiac troponin I (cTnI) detection system based on background fluorescence quenching of internal filtration effect (IFE) and study the influence of IFE on the sensitivity of cTnI detection. Three nanogold materials were synthesized as fluorescence quenchers, and rhodamine 6 G (R6G) and Cy5 were used as fluorescence probes. Six experimental systems were established to detect cTnI in negative serum test solutions and clinical serum samples. The sensitivity of each system was compared to explore the contribution of IFE to the detection sensitivity of cTnI. When applied to negative serum test solutions, the R6G-nanogold material I system exhibited a superior detection effect for cTnI, with a limit of detection (LOD) of 0.15 ng ml-1. When applied to clinical serum samples, the Cy5-nanogold material Ⅲ system yielded a better detection effect for cTnI, with the lowest concentration of cTnI detected at 2 ng ml-1. The first and second internal filtering effects in the proposed system can be achieved simultaneously, effectively avoiding light absorption interference from clinical serum samples and enhancing the sensitivity of the background fluorescence quenching detection of cTnI.

Combined detection of SARS-CoV-2 neutralizing antibodies and specific IgG in plasma based on SERS magnetic sensor.

In this study, a surface-enhanced Raman spectroscopy (SERS) magnetic sensor is established based on SERS principle and magnetic separation technology, and a highly sensitive, simple and fast method for quantitative detection of neutralizing antibodies (nABs) and specific IgG of SARS-CoV-2 in plasma is established combined with immunoassay. Two kinds of Raman nanospheres (RNPs) with different characteristic Raman shifts are used as signal sources and coupled to ACE2 and anti-IgG (FC) antibodies respectively, and magnetic beads are coupled to RBD. The competitive relationship between ACE2 and nABs, the binding relationship between specific IgG and anti-IgG (FC) antibodies are determined. The results show that the concentrations of nABs and specific IgG in the range of 10-2000 ng ml-1are well correlated with SERS response intensity, and the recoveries are both between 90%-110%, with good precision. Bilirubin and common anticoagulants have no interference on the detection results. This method is accurate, reliable, sensitive and does not require complex pre-treatment, and is expected to be used for simultaneous detection of nABs and specific IgG in plasma of SARS-CoV-2. It has guiding significance for the development and evaluation of vaccines and the formulation of individualized vaccination schedule.

Blockade of PD-1 and LAG-3 expression on CD8+ T cells promotes the tumoricidal effects of CD8+ T cells.

Background: The diffuse large B-cell lymphoma (DLBCL) has the highest incidence of all lymphomas worldwide. To investigate the functions of lymphocyte activation gene 3 (LAG-3) and programmed cell death 1 (PD-1) in tissues and peripheral blood of patients with DLBCL, the expression of LAG-3 and PD-1 genes in DLBCL-TCGA were analyzed. Methods: LAG-3 and PD-1 mRNA levels in DLBCL were analyzed using data from The Cancer Genome Atlas (TCGA) database. Utilize the Genotype-Tissue Expression (GTEx) database for assessing the variance in the expression of LAG-3, PD-1, and other associated factors between the tissues of DLBCL patients and healthy individuals. Immunohistochemistry was applied to detect the expression of LAG-3 and PD-1 levels in 137 cases of DLBCL tissues and 20 cases of reactive lymphoid hyperplasia. The prognostic value of LAG-3 and PD-1 were assessed using the Kaplan-Meier curve. The Estimation of Stromal and Immune cells in Malignant Tumor tissues using Expression data (ESTIMATE) and ssGSEA algorithm were used to explore the immune microenvironment of DLBCL. Additionally, the expression and co-expression of LAG-3 and PD-1 were detected on CD4 and CD8 T cells in peripheral blood samples from 100 cases of DLBCL tissues and 30 cases of healthy individuals using flow cytometry. Results: According to TCGA database, LAG-3 and PD-1 gene expression levels were significantly up-regulated in DLBCL tissues. LAG-3 and PD-1 levels were also strongly positively correlated with those of most infiltrating immune cells. Overall survival of patients with high LAG-3 and PD-1 co-expression was significantly shorter than that of patients with low co-expression. In DLBCL patients, LAG-3 and PD-1 were highly expressed in peripheral blood CD8+ T cells. In addition, LAG-3 was highly expressed in CD4+ T cells, while the expression of PD-1 in CD4+ T cells of DLBCL patients showed no significant difference compared to healthy individuals. Additionally, CD8+ T cells and SU-DHL6/OCI-LY3 from patients with DLBCL were co-cultured in vitro; after addition of LAG-3 and/or PD-1 inhibitors alone, an increased perforin and granzyme B secretion levels by CD8+ T cells were detected, as well as an increase in the overall proportion of tumor cells undergoing apoptosis. Conclusion: High LAG-3 and PD-1 levels significantly inhibit CD8+ T cell function, resulting in weakened ability to kill tumor cells. Combined LAG-3 and PD-1 blockade can restore CD8+ T cell function and provides a potential avenue for development of personalized cellular immunotherapy for DLBCL.

n-Type doping of BiVO4 with different F-doped concentrations for improving the electronic character of BiVO4 as a photoanode nanomaterial for solar water splitting: a first-principles study.

Atom doping has been realized as an effective way to improve the photocatalytic performance of the most promising photoanode material, BiVO4, but the effects of doping mass concentration still need to be explored. In this work, the effects of F-doping with different doping mass concentrations (1%, 2%, 5%, 10%, 15%, and 20%) on the electronic character of BiVO4 were examined theoretically using density functional theory (DFT) calculations. The thermal stability of BiVO4 with different F-doped mass concentrations was confirmed using formation energy (Ef) calculations though F-doped BiVO4 becomes harder as the mass concentration of induced dopants increases. n-Type doping effects on the electronic character of BiVO4 were observed upon F-doping, leading to the energy level of CBM shifting far away from the Fermi level and giving F-doped BiVO4 metallic character. Moreover, a linear relationship between the frontier energy level shifts and the total charge transfer amounts from doped F atoms to other atoms involved in F-doped BiVO4 was observed, which means the oxidizing capacity of the VBM is increased and the reducing capacity of the CBM is decreased upon increased F-doped mass concentration. Moreover, the recombination of photogenerated electron-hole pairs is suppressed by F-doping strategies, which will not change a lot with the increased F-doped mass concentration. This means atom doping is an effective strategy to improve the photocatalytic efficiency of the BiVO4, but the number of atoms introduced into BiVO4 should be appropriate.

Development of tetracycline-modified nanoparticles for bone-targeted delivery of anti-tubercular drug.

Background: Since the poor response to existing anti-tuberculosis drugs and low drug concentration in local bone tissues, the traditional drug therapy does not result in satisfactory treatment of osteoarticular tuberculosis. Thus, we report a rifapentine release system with imparted bone targeting potential using tetracycline (TC) -modified nanoparticles (NPs). Methods: TC was conjugated to PLGA-PEG copolymer via a DCC/NHS technique. Rifapentine-loaded NPs were prepared by premix membrane emulsification technique. The resulting NPs were characterized in terms of physicochemical characterization, hemolytic study, cytotoxicity, bone mineral binding ability, in vitro drug release, stability test and antitubercular activity. The pharmacokinetic and biodistribution studies were also performed in mice. Results: Rifapentine loaded TC-PLGA-PEG NPs were proved to be 48.8 nm in size with encapsulation efficiency and drug loading of 83.3% ± 5.5% and 8.1% ± 0.4%, respectively. The release of rifapentine from NPs could be maintained for more than 60 h. Most (68.0%) TC-PLGA-PEG NPs could bind to HAp powder in vitro. The cellular studies revealed that NPs were safe for intravenous administration. In vivo evaluations also revealed that the drug concentration of bone tissue in TC-PLGA-PEG group was significantly higher than that in other groups at all time (p < 0.05). Both NPs could improve pharmacokinetic parameters without evident organ toxicity. The minimal inhibitory concentration of NPs was 0.094 µg/mL, whereas this of free rifapentine was 0.25 µg/mL. Conclusion: Rifapentine loaded TC-PLGA-PEG NPs could increase the amount of rifapentine in bone tissue, prolong drug release in systemic circulation, enhance anti-tuberculosis activity, and thereby reducing dose and frequency of drug therapy for osteoarticular tuberculosis.

[Advances in cardiovascular protection effects and mechanisms of allicin].

Cardiovascular diseases(CVD) with high morbidity and mortality pose severe threats to human life. Allicin, a main active ingredient of garlic, possesses multiple pharmaceutical activities. It not only exerts cardioprotective effects but also prevents the risk factors for CVD. Allicin exerts cardioprotective effects via a variety of mechanisms, including inhibiting oxidative stress, apoptosis, autophagy, and inflammatory responses, regulating lipid metabolism and gut microbiota, inducing hydrogen sulfide production, and dilating vessels. Despite the valuable cardioprotective effects, the instability of allicin has hindered the basic research and clinical application. This paper reviews the progress in the cardioprotective effects and mechanisms of allicin in the last decade and summarizes the methods to improve the stability of allicin. In addition, this review provides a reference for further research and development of allicin in cardiovascular protection.

Facile modification of nanochitin in aqueous media for stabilizing tea tree oil based Pickering emulsion with prolonged antibacterial performance.

Nanochitins have been explored for preparing Pickering Emulsions, however its application is restricted by its simplex disperse nature. It was hypothesized that zwitterionic nanochitins should be capable of stabilizing oil/water (O/W) interfaces in wider pH range. Furthermore, the control of their size, disperse nature and self-assembly performance suggest the formulation of tunable emulsions. Zwitterionic nanochitins were prepared via Schiff base reaction. A systematic study was performed analyzing the disperse nature, fibril morphology, surface characteristic of modified nanochitins. Oil-in-Water Pickering Emulsions stabilized by modified nanochitins were formulated and emulsion stability was analyzed as function of concentration, pH and self-assembly property and further applied for prolonged antibacterial applications. Comparing freshly prepared nanochitins, neutral/alkaline stably dispersed nanochitins can be prepared while maintaining fibril characteristics such as fibril size, crystallinity, thermal stability and so on. Better suspension stability of modified nanochitins under alkaline conditon together with the self assembly performance resulting from amino groups and carboxyl groups benefit the enhanced emulsion stability under nanochitins concentreation of 0.2 %. Encapsulation of tea tree oil in Pickering Emulsions prolongs the diffusion rate oil in the aqueous environment, thus resulting prolongs its antibacterial performance against E. coli and B. subtilis.

Exome sequencing analysis of gastric primary myeloid sarcoma with monocytic differentiation with altered immunophenotype after chemotherapy: case report.

BACKGROUND: Myeloid Sarcoma with monocytic differentiation is rare and quite likely is missed by surgical pathologists. However it is frequently misdiagnosed because of its non-specific imaging and histological pattern. CASE PRESENTATION: We report the case of a 64-year-old woman with gastric primary myeloid sarcoma with monocytic differentiatio. Upper endoscopy revealed a neoplastic growth at the junction of the lesser curvature and gastric antrum. Except for a slightly increased peripheral monocyte count, no abnormalities were found on hematological and bone-marrow examination. Gastroscopic biopsy showed poorly differentiated atypical large cells with visible nucleoli and nuclear fission. Immunohistochemistry showed positive CD34, CD4, CD43, and CD56 expression, and weakly positive lysozyme expression. Immune markers for poorly differentiated adenocarcinoma, malignant melanoma, and lymphohematopoietic-system tumors were negative. The final diagnosis was myeloid sarcoma with monocytic differentiation. Chemotherapy did not shrink the tumor, so, radical surgery was performed. Although the tumor morphology did not change postoperatively, the immunophenotype did. CD68 and lysozyme expression (tumor tissue markers) changed from negative and weakly positive to strongly positive, AE1/3 expression (epithelial marker) changed from negative to positive, and CD34, CD4, CD43, and CD56 expression (common in naive hematopoietic cell-derived tumors) was greatly attenuated. Exome sequencing revealed missense mutations in FLT3 and PTPRB, which are associated with myeloid sarcoma, and in TP53, CD44, CD19, LTK, NOTCH2, and CNTN2, which are associated with lymphohematopoietic tumors and poorly differentiated cancers. CONCLUSION: We diagnosed myeloid sarcoma with monocytic differentiation after excluding poorly differentiated adenocarcinoma, common lymphohematopoietic-system tumors, epithelioid sarcoma, and malignant melanoma. We identified that the immunophenotypic of patient had alterations after chemotherapy, and FLT3 gene mutations. We hope that the above results will improve our understanding of this rare tumor.

Vitamin D3 alleviates inflammation in ulcerative colitis by activating the VDR-NLRP6 signaling pathway.

Inflammation is a key factor in the development of ulcerative colitis (UC). 1,25-dihydroxyvitamin D3 (1,25(OH)2D3, VD3), as the major active ingredient of vitamin D and an anti-inflammatory activator, is closely related to the initiation and development of UC, but its regulatory mechanism remains unclear. In this study, we carried out histological and physiological analyses in UC patients and UC mice. RNA sequencing (RNA-seq), assays for transposase-accessible chromatin with high-throughput sequencing (ATAC-seq), chromatin immunoprecipitation (ChIP) assays and protein and mRNA expression were performed to analyze and identify the potential molecular mechanism in UC mice and lipopolysaccharide (LPS)-induced mouse intestinal epithelial cells (MIECs). Moreover, we established nucleotide-binding oligomerization domain (NOD)-like receptor protein nlrp6 -/- mice and siRNA-NLRP6 MIECs to further characterize the role of NLRP6 in anti-inflammation of VD3. Our study revealed that VD3 abolished NOD-like receptor protein 6 (NLRP6) inflammasome activation, suppressing NLRP6, apoptosis-associated speck-like protein (ASC) and Caspase-1 levels via the vitamin D receptor (VDR). ChIP and ATAC-seq showed that VDR transcriptionally repressed NLRP6 by binding to vitamin D response elements (VDREs) in the promoter of NLRP6, impairing UC development. Importantly, VD3 had both preventive and therapeutic effects on the UC mouse model via inhibition of NLRP6 inflammasome activation. Our results demonstrated that VD3 substantially represses inflammation and the development of UC in vivo. These findings reveal a new mechanism by which VD3 affects inflammation in UC by regulating the expression of NLRP6 and show the potential clinical use of VD3 in autoimmune syndromes or other NLRP6 inflammasome-driven inflammatory diseases.

Study on the performance of novel nanomaterials for detection of biomarkers such as PCT based on immunochromatography sensitivity.

In this study, by comparing the UV-vis spectral characteristics of colloidal gold and colloidal gold enhancer, and their differences as immunochromatographic tracers in the qualitative detection of PCT, IL-6, Hp and quantitative determination of PCT performance, the factors that may affect the sensitivity were discussed. The results show that the absorbance at 520 nm of CGE diluted 20-fold and colloidal gold diluted 2-fold were comparable, and the sensitivity of CGE immunoprobe for qualitative detection of PCT, IL-6 and Hp was higher than that of colloidal gold immunoprobe, and the reproducibility and accuracy of both immunoprobes for quantitative detection of PCT were good. Indicating that the high sensitivity of CGE immunoprobe detection is mainly due to the absorption coefficient of CGE at 520 nm is about 10 times that of colloidal gold immunoprobe, CGE has stronger light absorption capacity and stronger quenching effect on rhodamine 6 G on the nitrocellulose membrane surface of the test strip.

Allicin ameliorates imiquimod-induced psoriasis-like skin inflammation via disturbing the interaction of keratinocytes with IL-17A.

BACKGROUND AND PURPOSE: Psoriasis is an inflammatory skin disease of chronic recurrence mediated by the interaction between IL-17 and keratinocytes, which sustains a vicious circle of inflammation. Safe and effective natural medicine is a potential strategy for the clinical treatment of psoriasis. Given its prominent anti-proliferative and anti-inflammatory properties, we investigated the actions of allicin in improving psoriasis. EXPERIMENTAL APPROACH: Pharmacodynamic studies were carried out in mice after topical administration of allicin against psoriasis-like lesions induced by imiquimod. Skin sensitization tests were evaluated on guinea pigs. Toxicological studies and skin irritation tests were assessed by consecutive topical allicin alone on the skin of rabbits. RNA-seq probed transcriptomic changes following allicin. Western blot explored the actions of allicin on the interaction between IL-17A and keratinocytes. Changes in inflammatory factor expression were analysed by qPCR and immunohistochemistry. KEY RESULTS: Allicin significantly improved the epidermal structure by inhibiting the excessive proliferation and reduced apoptosis of keratinocytes. Furthermore, allicin reduced the secretion of inflammatory cytokines (IL-17A/F, IL-22, IL-12, and IL-20), chemokines (CXCL2, CXCL5, and CCL20), and anti-bacterial peptides (S100a8/9). Mechanistically, allicin directly inhibited the IL-17-induced TRAF6/MAPK/NF-κB and STAT3/NF-κB signalling cascades in keratinocytes, thus breaking the positive inflammatory feedback and alleviating imiquimod-induced psoriasis-like dermatitis in mice. Importantly, topical administration of allicin did not cause skin allergy, and the safety and adaptability of long-term application were verified. CONCLUSIONS AND IMPLICATIONS: Interfering with IL-17 signalling in keratinocytes with allicin is a promising strategy for treating psoriasis, given its safety and effectiveness.

Allicin shows antifungal efficacy against Cryptococcus neoformans by blocking the fungal cell membrane.

Allicin, which is generated by the catalytic reaction between alliin and alliinase extracted from garlic, has been shown to have a wide range of antimicrobial activities, but its anti-Cryptococcus efficacy and mechanism are not quite clear. Here, we have determined that the Conversion rate of allicin in the reaction product reached 97.5%. The minimal inhibitory concentration (MIC) of allicin against Cryptococcus neoformans (C. neoformans) H99 was 2 µg/ml, which is comparable to fluconazole (FLU, 1 µg/ml). Furthermore, allicin exhibited effective antifungal activity against 46 clinical isolates of C. neoformans, and the MICs ranged from 1 to 8 µg/ml, even for AmB-insensitive strains. Interestingly, allicin also exerted additive or synergistic effects when combined with amphotericin B (AmB) and FLU. Time-killing curves and long-term live cell imaging of H99 showed that 4 MIC of allicin had fungicide activity. Additionally, allicin (4 and 8 mg/kg) exerted a dose-dependent therapeutic effect on H99-infected mice by significantly reducing the wet pulmonary coefficient and Cryptococcus load and reducing lung damage. Even the efficacy of 8 mg/kg was comparable to FLU (20 mg/kg). Transcriptomics revealed that allicin may act on the cell membrane of H99. Subsequently, transmission electron microscopy (TEM) observations showed that allicin clearly breached the cell membrane and organelles of H99. Confocal laser scanning microscopy (CLSM) results further confirmed that allicin disrupted the permeability of the cell membranes of H99 in a dose-dependent manner. Allicin exhibits strong anti-C. neoformans activity in vitro and in vivo, mainly by destroying the permeability and related functions of Cryptococcus cell membranes.

Expression of PD-L1 and YWHAZ in Patients with Diffuse Large B Cell Lymphoma: A Possible Association with the Prognosis of Lymphoma.

Diffuse large B cell lymphoma (DLBCL) is the most common pathological subtype of non-Hodgkin lymphoma (NHL) and is the most common type of adult lymphoma. Due to the poor prognosis of relapsed/refractory DLBCL, new drug targets and therapeutic methods are urgently needed. We investigated the expression of programmed death ligand 1 (PD-L1) and 3-monooxygenase/tryptophan 5-monooxygenase activating protein zeta (14-3-3ζ or YWHAZ) in patients with DLBCL. The purpose was to verify the expression levels of YWHAZ and PD-L1 and their relationships with the prognosis of DLBCL and to lay a foundation for further study on the role of YWHAZ and PD-L1 in DLBCL. Immunohistochemistry was used in 140 patients with DLBCL to test protein expression levels of YWHAZ and PD-L1. All patients were followed up in the hospital or by telephone or via WeChat. The positive expression rate of YWHAZ was 62.14% (87/140). The expression was negatively correlated with the positive expression of BAD (r = -0.177, P = 0.036) and positively correlated with the positive expression of BCL-2 (r = 0.180, P = 0.033). When the cut-off value for PD-L1 was established at 5%, 10%, 15%, and 20%, the corresponding positive expression rates of PD-L1 were 79.66% (94/118), 51.69% (61/118), 40.68% (48/118), and 36.44% (43/118). YWHAZ significantly affected the OS of DLBCL (P ≤ 0.001). The prognosis of the patients was related to the positive expression of PD-L1 when the cut-off value of PD-L1 was 5% (P = 0.033). However, positive expression of PD-L1 was not associated with the prognosis when the cut-off values of PD-L1 were 10% (P = 0.404), 15% (P = 0.208), and 20% (P = 0.408). The positive expression of YWHAZ (hazard ratio 6.215; 95% confidence interval 3.214-12.017; P < 0.05) was an independent adverse prognostic factor for OS. YWHAZ may be an important oncogene in the occurrence and development of DLBCL and may be used as a therapeutic target. PD-L1 may be an oncogene or tumor suppressor gene in the occurrence and development of DLBCL. Different cut-off values of PD-L1 may affect the prognosis of DLBCL.

Correlation study of PD-L1, CD4, CD8, and PD-1 in primary diffuse large B-cell lymphoma of the central nervous system.

We investigated the clinicopathological role of the PD-1/PD-L1 pathway in the primary diffuse large B-cell lymphoma of the central nervous system (PCNS-DLBCL). Standardized staining for PD-L1 was performed by machine staining, and internationally accepted interpretation methods were used. The PD-L1 immunostaining ≥ 20 % of all cells in slices was defined as high expression of PD-L1. CD4, CD8, and PD-1 tumor-infiltrating lymphocytes (TILs) were enumerated, and the median was defined as the cutoff value. Values higher than the median was defined as high expression. Thirty-four cases (64.2 %) showed high expression of PD-L1. PD-L1 expression was associated with a good prognosis when 20 % was considered as cutoff value and had the smallest P value. By contrast, a low number of CD8+ or PD-1+ TILs was associated with poor prognosis. Patients with low expression of PD-L1 had poor overall survival (P = 0.001), and those with increased CD8 or PD-1 TILs tended to have improved overall survival (P = 0.004 and 0.024, respectively). Low number of monocytes, increased number of lymphocytes, IPI score ≥ 2, ECOG PS ≥ 2, LDH ≥ 250, and Ki67 ≥ 70 % were independent prognostic factors for OS. In conclusion, PD-L1 expression, CD8 and PD-1 TILs, monocyte status, and ECOG PS might be prognostic markers and therapeutic targets of PCNS-DLBCL.

Effects of hsa-miR-28-5p on Adriamycin Sensitivity in Diffuse Large B-Cell Lymphoma.

Background: Adriamycin (doxorubicin) is an important traditional drug that exhibits cytotoxicity in Diffuse Large B-cell Lymphoma (DLBCL). Doxorubicin affects the DLBCL cells at all stages of their cell cycle. Combined with our previous results, this study discovered that the overexpression of hsa-miR-28-5p inhibited the proliferation, promoted apoptosis, and triggered cell cycle arrest at the S-phase in DLBCL cells. However, the effect of (Homo sapiens, hsa)-microRNA (miR)-28-5p on doxorubicin sensitivity in DLBCL has not been investigated. This study aims to reveal the effects of hsa-miR-28-5p on doxorubicin sensitivity at the level of DLBCL cells. Methods: To determine the optimal concentration of doxorubicin, different concentrations of doxorubicin were used to treat DLBCL cells. CCK-8 assay was used to detect the proliferation of DLBCL cells. The hsa-miR-28-5p-mimic NC and hsa-miR-28-5p mimic were transfected to doxorubicin-mediated DLBCL cells. Simultaneously, blank control groups were set up. The cells were cultured and transfected for 24 h. Next, each group was administered with different concentrations of doxorubicin and cultured again for 24 h to observe the effects of hsa-miR-28-5p on doxorubicin sensitivity at different times. The proliferation, early apoptosis, and late apoptosis in DLBCL cells were determined using soft agar colony-forming assay, mitochondrial membrane potential assay, and caspase-3 activity assay, respectively. The apoptosis and cell cycle were explored using Annexin V-PE/7-AAD and PI/RNase staining buffer, respectively. We speculated that PD-L1 might be involved in the effect of hsa-miR-28-5p on the sensitivity of adriamycin (doxorubicin) in the DLBCL cells. Hence, we performed immunohistochemistry (IHC) to determine PD-L1 expression within formalin-fixed paraffin-embedded (FFPE) samples from 52 DLBCL cases. Results: The optimal concentration of doxorubicin targeting DLBCL cells was found to be 3.028 µmol/l. The effect of doxorubicin on DLBCL cells was time- and concentration-dependent. hsa-miR-28-5p mimic + doxorubicin remarkably decreased proliferation of DLBCL. DLBCL cell apoptosis rate was the highest in hsa-miR-28-5p mimic + doxorubicin group. Apart from that, hsa-miR-28-5p mimic plus doxorubicin had the best effect in promoting DLBCL cell apoptosis. After the intervention of hsa-miR-28-5p mimic + doxorubicin on DLBCL cells, the cell cycle was arrested in the S-phase and DNA synthesis was blocked. hsa-miR-28-5p mimic + doxorubicin could regulate the cycle of DLBCL cells. As a result, overexpression of hsa-miR-28-5p combined with doxorubicin is possibly involved in the development of DLBCL by affecting the proliferation, apoptosis, and cycle of DLBCL cells. PD-L1 showed an association with the prognosis of DLBCL patients. Combining with the literature, this suggested hsa-miR-28-5p may influence DLBCL occurrence and therapeutic effect by regulating the PD-L1 level. Conclusion: The combination of hsa-miR-28-5p mimic and doxorubicin may be considered more effective in inhibiting growth, arresting the cell cycle, and promoting cell apoptosis of DLBCL cells compared to using doxorubicin alone. The effects of doxorubicin on DLBCL cells were found to be time- and concentration-dependent. The overexpression of hsa-miR-28-5p enhanced the effect of doxorubicin on DLBCL cells, which may be attributed to the regulation of PD-L1 levels.

Construction of Prediction Model of Radiotherapy Set-Up Errors in Patients with Lung Cancer.

Objective: This study intends to construct an error distribution prediction model and analyze its parameters and analyzes the boundary size of CTV extension to PTV, so as to provide a reference for lung cancer patients to control clinical set-up errors and radiotherapy planning. Methods: The prior SBRT set-up error data of 50 patients with lung cancer treated by medical linear accelerator were selected, the Gaussian mixture model was adopted to construct the error distribution prediction model, and the model parameters were solved, based on which the emission boundary from CTV to PTV was calculated. Results: According to the analysis of the model parameters, the spatial distribution of set-up errors is mainly concentrated in the direction of four central points (µ 1 ~ µ 4), and the error is smaller in the Vrt direction (-0.991~2.808 mm) and Lat direction (-0.447~1.337 mm) and larger in the Lng direction (-1.065~4,463 mm). The possibility of offset of set-up errors in µ 2 and µ 3 direction (0.4440, 02198) is greater than that of µ 1 and µ 4 (0.1767, 0.1595). The standard deviation of set-up errors can reach 0.538 mm. The theoretical expansion boundary of CTV to PTV in Vrt, Lng, and Lat can be calculated as 1.7963 mm, 2.3749 mm, and 0.6066 mm. Conclusion: The GMM Gaussian mixture model can quantitatively describe and predict the set-up errors distribution of lung cancer patients and can obtain the emission boundary of CTV to PTV, which provides a reference for radiotherapy set-up errors control and tumor planning target expansion of lung cancer patients without SBRT.