The Expression and Biological Significance of PD-L1 on Lung Cancer Cell Lines

Background and objective Tumor-associated PD-L1 expression was recently shown to promote T-cell apoptosis and proposed as a potential mechanism of immune evasion by tumors. On the basis of the ability of tumor-associated PD-L1 to mediate activated T-cell death, it is likely that manipulation of the PD-L1 pathway at defined time points during the development of the T-cell antitumor immune response can enhance the efficacy of T-cell-based immunotherapy. Here, the levels of expression of PD-L1 on lung cancer cell lines and its role in interaction of CTL and target cells was investigated. Methods Human PBMC derived DCs were loaded with apoptotic tumor cells and stimulated by CD40 mAb (5C11). Tumor specific CTL was generated in vitro by autologous T cells co-cultured with mature DCs. Expression of PD-L1 on lung cancer cell lines H1299 and A549 were analyzed by FCM. JAM assay was used to detect the cytolytic activity of CTL with or without blocking PD-L1 by PD-L1 mAb respectively. The concentrations of IFN-γ in supernatants from distinct groups were analyzed by ELISA. Results Tumor cells-loaded mature DCs could induce the generation of the tumor specific CTL. Expression of PD-L1 was low on A549 cell, but high on H1299 cell. Blockade of PD-L1 on A549 could not improve cytolytic effect of CTL on target cells and IFN-γ production, but fragmentation of H1299 cells and IFN-γ production were significantly enhanced by the combination of PD-L1 mAb and CTL. Conclusion Expression of PD-L1 on lung cancer cell line can decrease the cytolytic effect of CTL on target cells

Genome-wide analysis of the Catalpa bungei caffeic acid O-methyltransferase (COMT) gene family: identification and expression profiles in normal, tension, and opposite wood

Caffeic acid O-methyltransferase (COMT) is an important protein that participates in lignin synthesis and is associated with the ratio of G-/S-type lignin in plants. COMTs are associated with the wood properties of forest trees; however, little known about the COMT family in Catalpa bungei, a valuable timber tree species in China . We performed a comprehensive analysis of COMT genes in the C. bungei genome by describing the gene structure and phylogenetic relationships of each family member using bioinformatics-based methods. A total of 23 putative COMT genes were identified using the conserved domain sequences and amino acid sequences of COMTs from Arabidopsis thaliana and Populus trichocarpa as probes. Phylogenetic analysis showed that 23 CbuCOMTs can be divided into three groups based on their structural characteristics; five conserved domains were found in the COMT family. Promoter analysis indicated that the CbuCOMT promoters included various cis-acting elements related to growth and development. Real-time quantitative polymerase chain reaction (PCR) analysis showed differential expression among CbuCOMTs. CbuCOMT2, 7, 8, 9, 10, 12, 13, 14, 21, and 23 were mainly expressed in xylem. Only CbuCOMT23 was significantly downregulated in tension wood and upregulated in opposite wood compared to normal wood. Our study provides new information about the CbuCOMT gene family and will facilitate functional characterisation in further research

The interaction between the soluble programmed death ligand-1 (sPD-L1) and PD-1+ regulator B cells mediates immunosuppression in triple-negative breast cancer

Accumulating evidence suggests that regulatory B cells (Bregs) play important roles in inhibiting the immune response in tumors. Programmed death 1 (PD-1) and programmed death ligand 1 (PD-L1) are important molecules that maintain the balance of the immune response and immune tolerance. This study aims to evaluate the soluble form of PD-L1 and its function in inducing the differentiation of B lymphocytes, investigate the relationship between soluble PD-L1 (sPD-L1) and B-cell subsets, and explore the antitumor activity of T lymphocytes after PD-L1 blockade in coculture systems. In an effort to explore the role of sPD-L1 in human breast cancer etiology, we examined the levels of sPD-L1 and interleukin-10 (IL-10) in the serum of breast tumor patients and the proportions of B cells, PD-1+ B cells, Bregs, and PD-1+ Bregs in the peripheral blood of patients with breast tumors and assessed their relationship among sPD-L1, IL-10, and B-cell subsets. The levels of sPD-L1 and IL-10 in serum were found to be significantly higher in invasive breast cancer (IBCa) patients than in breast fibroadenoma (FIBma) patients. Meanwhile, the proportions and absolute numbers of Bregs and PD-1+ Bregs in the peripheral blood of IBCa patients were significantly higher than those of FIBma patients. Notably, they were the highest in triple-negative breast cancer (TNBC) among other subtypes of IBCa. Positive correlations of sPD-L1 and IL-10, IL-10 and PD-1+ Bregs, and also sPD-L1 and PD-1+ Bregs were observed in IBCa. We further demonstrated that sPD-L1 could induce Breg differentiation, IL-10 secretion, and IL-10 mRNA expression in a dose-dependent manner in vitro. Finally, the induction of regulatory T cells (Tregs) by Bregs was further shown to suppress the antitumor response and that PD-L1 blockade therapies could promote the apoptosis of tumor cells. Together, these results indicated that sPD-L1 could mediate the differentiation of Bregs, expand CD4+ Tregs and weaken the antitumor activity of CD4+ T cells. PD-L1/PD-1 blockade therapies might be a powerful therapeutic strategy for IBCa patients, particularly for TNBC patients with high level of PD-1+ Bregs

Sub-Bowman keratomileusis versus femtosecond laser in situ keratomileusis on the long-term visual recovery: A comparative study

Purpose: To explore the long-term visual quality of the same subjects after sub-Bowman keratomileusis (SBK) or femtosecond laser in situ keratomileusis (FS-LASIK). Methods: This prospective study included patients screened for corneal refractive surgery at the Refractive Surgery Center of our Hospital between November 2017 and March 2018. One eye underwent SBK, while the other eye underwent FS-LASIK. Total higher-order aberrations, coma aberrations, and clover aberrations were evaluated before and at 1 month and 3 years after the procedure. The visual satisfaction of both eyes was investigated, respectively. The participants completed a surgical satisfaction questionnaire. Results: Thirty-three patients were included. There were no significant differences in total higher-order aberrations, coma aberrations, and clover aberrations between the two procedures before and 1 month and 3 years after surgery (all P > 0.05), except for the total coma aberrations in FS-LASIK were significantly higher compared with the SBK group at 1 month after surgery [0.51 (0.18, 0.93) vs. 0.77 (0.40, 1.22), P = 0.019]. The surgical satisfaction questionnaire scores of the SBK group and the FS-LASIK group were 9.8 ± 0.8 and 9.8 ± 0.8, respectively, at 1 month, and 9.7 ± 0.9 and 9.7 ± 1.0, respectively, at 3 years (all P > 0.05). Conclusion: There were no differences in corneal aberrations and satisfaction between SBK and FS-LASIK procedures at 1 month and 3 years

Genetic Variation of Growth Traits and Genotype-by-Environment Interactions in Clones of Catalpa bungei and Catalpa fargesii f. duclouxii

Clones of Catalpa bungei and Catalpa fargesii f. duclouxii were studied over several years in central China to explore genetic variation in growth traits and to identify clones of high wood yield and high stability. The genetic parameters for height, diameter at breast height (DBH), and stem volume of clones, were estimated. The effect of clone × year on the increment of stem volume in the two species was analyzed by genotype and genotype × environment (GGE) biplot methods. Significant differences in growth traits among clones and between species were found. The growth of C. bungei exceeded that of C. fargesii f. duclouxii after 4 years. Furthermore, from the 5th year, the repeatability and genetic variation coefficient (GCV) of the C. bungei clones were higher than those of the C. fargesii f. duclouxii clones in most cases. The phenotypic variation coefficient (PCV) of the C. fargesii f. duclouxii clones was significantly lower than that of the C. bungei clones. The repeatability of stem volume was intermediate or high in the two species. ANOVA revealed significant effects of the clone by year interaction in these two species. GGE biplot analysis revealed that wood yield and stability were largely independent in C. bungei; clones 22-03, 19-27, and 20-01 were the optimal clones in this species. In contrast, the optimal clones 63 and 128 of C. fargesii f. duclouxii combined the desired characteristics of high yield and high stability. In conclusion, our results indicated that the height and stem volume of C. bungei was under strong genetic control, whereas that of C. fargesii f. duclouxii was influenced by the environment more than by genetic effects. Genetic improvement by clone selection can be expected to be effective, as the repeatability of stem volume was high. Francis and Kannenberg’s method and GGE biplot analysis were used in combination to evaluate the clones. C. bungei clone 22-03 and C. fargesii f. duclouxii clones 63 and 128 were identified as the optimal clones, which exhibited both a high increment of stem volume and high stability

Research into the correlation between positional skull deformation and motor performance of infants aged under 4 months

Abstract Objective To investigate the correlation between positional skull deformation (PD) and motor performance of infants under 4 months of age. Methods Infants aged under 4 months were enrolled in the children’s healthcare and the premature infants follow-up Clinic of the Second Affiliated Hospital of Army Military Medical University. The cranial vault asymmetry (CVA) and cephalic index (CI) were calculated in all infants, and the infant motor performance test (TIMP) was used to evaluate the infant motor performance. The motor performances of infants with different types and degrees of PD were compared, so were the incidences of PD in infants with different motor performance levels. Results Overall, 2118 infants were recruited and divided according to the types of PD and TIMP scores. The comparison of TIMP scores within different types of PD at different months of age showed that, regardless of the types of PD, TIMP scores of infants with PD were lower than those of normal infants. In particular, the difference in TIMP scores was statistically significant (P < 0.05) in infants with dolichocephaly, plagiocephaly,dolicho-plagiocephaly and brachy-plagiocephy. In addition, the comparison of CVA values of infants with different TIMP score levels at different months of age showed that the CVA values of the extremely low-level group were significantly higher than those of the medium-level and high-level group, especially in the 3-month-old and 4-month-old groups, which showed significant statistical differences (P < 0.05). Conclusions PD and motor performance of infants aged under 4 months seem to interact and influenc each other. The more serious the severity of PD were,the worse the motor performance of infants. Conversely, the incidence of PD increased in infants with poor motor performance

Liquid Chromatography–Tandem Mass Spectrometry Determination and Pharmacokinetic Analysis of Amentoflavone and Its Conjugated Metabolites in Rats

Amentoflavone (AMF) is a biflavone found in many herbal dietary supplements. To investigate the pharmacokinetic profile of AMF in rats, a sensitive, simple, and accurate liquid chromatography–tandem mass spectrometry (LC–MS/MS) method was developed and used to monitor AMF and its conjugated metabolites in plasma. AMF was administered to rats by oral gavage (po), or by intravenous (iv) or intraperitoneal (ip) injection. Plasma samples (with apiolin as an internal standard) were liquid/liquid extracted after hydrolysis with β-glucuronidase/sulfatase in vitro. Following chromatographic separation on a C18 column with a methanol:water:formic acid (70:30:0.1, v/v/v) mobile phase, AMF and internal standard were determined by electrospray ionization in negative ion mode and their precursor–product ion pairs (<i>m</i>/<i>z</i> 537.1 → 374.9 and <i>m</i>/<i>z</i> 269.2 → 224.9, respectively) were used for measurement. This bioanalytical method was fully validated and showed good linearity (<i>r</i>² > 0.99), wide dynamic range (0.93–930 nmol/L), and favorable accuracy and precision. After iv or ip AMF (10 mg/kg) injection, 73.2% ± 6.29% and 70.2% ± 5.18% of the total AMF detected in plasma was present as conjugated metabolites. Furthermore, AMF and AMF conjugates showed similar time courses with no significant differences in the time to reach the maximum plasma concentration (<i>t</i>_max) and terminal half-life (<i>t</i>_1/2) (<i>p</i> > 0.05). Following po AMF administration (300 mg/kg), 90.7% ± 8.3% of the total AMF was circulating as conjugated metabolites. When compared with iv administration (with dose correction), the bioavailability of po AMF was very low (0.04% ± 0.01% for free AMF; 0.16% ± 0.04% for conjugated AMF). Collectively, these data provided a preliminary pharmacokinetic profile for AMF that should inform further evaluations of its biological efficacy and preclinical development