RNA sequencing reveals dynamic expression of lncRNAs and mRNAs in caprine endometrial epithelial cells induced by Neospora caninum infection.

BACKGROUND: The effective transmission mode of Neospora caninum, with infection leading to reproductive failure in ruminants, is vertical transmission. The uterus is an important reproductive organ that forms the maternal-fetal interface. Neospora caninum can successfully invade and proliferate in the uterus, but the molecular mechanisms underlying epithelial-pathogen interactions remain unclear. Accumulating evidence suggests that host long noncoding RNAs (lncRNAs) play important roles in cellular molecular regulatory networks, with reports that these RNA molecules are closely related to the pathogenesis of apicomplexan parasites. However, the expression profiles of host lncRNAs during N. caninum infection has not been reported. METHODS: RNA sequencing (RNA-seq) analysis was used to investigate the expression profiles of messenger RNAs (mRNAs) and lncRNAs in caprine endometrial epithelial cells (EECs) infected with N. caninum for 24 h (TZ_24h) and 48 h (TZ_48 h), and the potential functions of differentially expressed (DE) lncRNAs were predicted by using Gene Ontology (GO) enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis of their mRNA targets. RESULTS: RNA-seq analysis identified 1280.15 M clean reads in 12 RNA samples, including six samples infected with N. caninum for 24 h (TZ1_24h-TZ3_24h) and 48 h (TZ1_48h-TZ3_48h), and six corresponding control samples (C1_24h-C3_24h and C1_48h-C3_48h). Within the categories TZ_24h-vs-C_24h, TZ_48h-vs-C_48h and TZ_48h-vs-TZ_24h, there were 934 (665 upregulated and 269 downregulated), 1238 (785 upregulated and 453 downregulated) and 489 (252 upregulated and 237 downregulated) DEmRNAs, respectively. GO enrichment and KEGG analysis revealed that these DEmRNAs were mainly involved in the regulation of host immune response (e.g. TNF signaling pathway, MAPK signaling pathway, transforming growth factor beta signaling pathway, AMPK signaling pathway, Toll-like receptor signaling pathway, NOD-like receptor signaling pathway), signaling molecules and interaction (e.g. cytokine-cytokine receptor interaction, cell adhesion molecules and ECM-receptor interaction). A total of 88 (59 upregulated and 29 downregulated), 129 (80 upregulated and 49 downregulated) and 32 (20 upregulated and 12 downregulated) DElncRNAs were found within the categories TZ_24h-vs-C_24h, TZ_48h-vs-C_48h and TZ_48h-vs-TZ_24h, respectively. Functional prediction indicated that these DElncRNAs would be involved in signal transduction (e.g. MAPK signaling pathway, PPAR signaling pathway, ErbB signaling pathway, calcium signaling pathway), neural transmission (e.g. GABAergic synapse, serotonergic synapse, cholinergic synapse), metabolism processes (e.g. glycosphingolipid biosynthesis-lacto and neolacto series, glycosaminoglycan biosynthesis-heparan sulfate/heparin) and signaling molecules and interaction (e.g. cytokine-cytokine receptor interaction, cell adhesion molecules and ECM-receptor interaction). CONCLUSIONS: This is the first investigation of global gene expression profiles of lncRNAs during N. caninum infection. The results provide valuable information for further studies of the roles of lncRNAs during N. caninum infection.

1H-NMR-guided isolation of enantiomeric coumarin-monoterpenes with anti-inflammatory activity from Gerbera piloselloides.

1H-NMR guided fractionation led to the isolation of twenty-two coumarin monoterpenes from the whole plant of Gerbera piloselloides, among which fourteen were undescribed. All coumarin monoterpenes were initially found to be racemates without optical activity. Subsequently, eleven pairs of optically pure enantiomers were successfully separated by chiral phase HPLC. Their structures and absolute configurations were unambiguously determined based on their spectroscopic data, calculated/experimental electronic circular dichroism (ECD) data, and X-ray diffraction analysis. Bioassays in LPS-treated RAW 264.7 cells revealed that the four compounds possessed moderate anti-inflammatory activity. In addition, the correlations between the cotton effect (CE) from δ-lactone at approximately 210-220 nm in CD spectra and γ-C or the ring fused at γ-C of the skeleton were reported for the first time.

The outcome of surgical and non-surgical treatments for traumatic optic neuropathy: a comparative study of 685 cases.

Background: The choice and efficacy of surgical or/and surgical treatments for traumatic optic neuropathy (TON) remained controversial by now. This study aims to present the outcomes of surgical and nonsurgical treatments for TON in our center. Methods: A total of 685 consecutive patients were retrospectively included in the study. And divided into surgical and non-surgical groups. All cases were treated with corticosteroids for 3 days after admission. Endoscopic optic decompression was applied to 479 patients of surgical group; The other 206 patients of nonsurgical were administered with corticosteroids alone. The visual outcomes before and after treatment were compared with Wilcoxon rank and tests. The improvement rate between two groups were compared with chi-square test. Results: The visual acuity (VA) after treatment was significantly better than that before treatment (P=0.000). Overall VA improvement rate in the surgical group was better than that in non-surgical group (42.8% vs. 35.4%) with no significant difference (P=0.072). The VA improvement rate was significant greater in the surgical group than that in the non-surgical group in the patients with NLP before treatment (P=0.028). The VA improvement rate was better in the surgical group than that in the non-surgical group (71.9% vs. 57.8%) but with no significant difference. The final overall VA was 0.1 or better in 43 cases; 104 cases were able to count fingers; hand motion (HM) became perceivable in 132 cases; light perception (LP) was achieved in 53 cases; and no light perception (NLP) remained in 353 cases. Conclusions: Endoscopic optic nerve decompression (EOND) combined with corticosteroids or corticosteroids alone could reach the improvement for patients with TON. The EOND combined with corticosteroids could achieve better VA improvement in patients with NLP.

Silencing Tautomerization to Isolate Unstable Physalins from Physalis minima.

The inherent structural instability of some physalins has hampered the isolation and identification of these compounds for approximately 50 years, and an effective method to overcome these challenges remains unavailable. In the present study, the unprecedented tautomerization mechanism of unstable physalins was elucidated by performing isotopic labeling experiments and DFT calculations, which led to the successful separation of tautomers and isolation of highly pure products for the first time. As a result, 15 new physalins, physaminins A-O (1-15), as well as 17 known analogues (16-32), were isolated from the whole plants of Physalis minima L. The chemical structures of the new compounds were established by performing a comprehensive analysis of spectroscopic data, and their absolute configurations were confirmed by using computational ECD calculations and/or single-crystal X-ray diffraction analyses. All obtained isolates were evaluated for their antiproliferative effects against four human cancer cell lines (A549, HepG2, MCF-7, and SCG-7901) and two noncancerous cell lines (RAW 264.7 and human normal hepatocytes L02), as well as their anti-inflammatory activities by measuring their abilities to inhibit NO production in LPS-stimulated murine RAW 264.7 cells in vitro. Compounds 1-5, 13, 16, 18, 19, 23, and 30 exerted significant antiproliferative effects on the four human cancer lines, with IC50 values ranging from 0.2(0) to 24.7(2) µM, and these compounds were not toxic to the two noncancerous cell lines at a concentration of 10 µM. Moreover, compounds 7, 10, 11, 12, 14, 17, 22, and 27 significantly inhibited NO production, with IC50 values ranging from 2.9(1) to 9.5(2) µM.

Sex- and size-dependent accumulation of Dechlorane Plus flame retardant in a wild frog-eating snake Amphiesma stolata.

Despite several studies having addressed the bioaccumulation of Dechlorane Plus (DP) flame retardant in wildlife, there is still a dearth of information for reptiles in general and for snakes in particular. Here, we report the residue levels and trophic transfer of syn-, anti-, and anti-Cl11-DP in a frog-eating snake-namely, the striped keelback snake Amphiesma stolata-from a DP hotspot in South China. The concentrations of syn-, anti-, and anti-Cl11-DP in A. stolata ranged from 1.06-21.2, 2.13-21.5, and 0.16-10.6 ng/g lipid weight, respectively, with significantly higher levels in males compared with females. Statistical analysis showed that the concentrations of these chemicals were negatively correlated with body sizes (length and mass) of the snake. The fractional abundance of anti-DP (fanti) did not significantly differ either between the sexes or between A. stolata and its diet (i.e., frogs). However, fanti showed positive correlations with the snake's body size and negative correlations with ∑DP concentrations (summed concentrations of syn- and anti-DP), indicating that body size and DP residue levels are important factors influencing DP isomeric profiles in these snakes. Biomagnification factors estimated based on the relationship between A. stolata and frogs were 0.49 ± 0.01 (mean ± SE), 0.44 ± 0.09, and 1.79 ± 0.54 for syn-, anti-, and anti-Cl11-DP, respectively, suggesting trophic dilution of syn- and anti-DP and a mild biomagnification of anti-Cl11-DP from frogs to snakes.

Bioaccumulation and maternal transfer of two understudied DDT metabolites in wild fish species.

While the secondary metabolites of DDT such as 2,2-bis(chlorophenyl)-1-chloroethylene (DDMU) and 2,2-bis(chlorophenyl)methane (DDM) have been detected in the environment for several decades, knowledge is extremely limited on their bioaccumulation characteristics. Here, we reported the bioaccumulation and maternal transfer of p,p'-DDMU and p,p'-DDM in two wild fishes, i.e., the northern snakehead (Channa argus) and crucian carp (Carassius auratus), from a DDT contaminated site in South China. The hepatic concentrations of p,p'-DDMU and p,p'-DDM in the fish were up to 549 and 893 ng/g lipid weight, contributing 5.3% and 3.2% in average to ΣDDXs (the sum concentrations of DDT and its 6 metabolites), respectively. The residues of p,p'-DDMU and p,p'-DDM in the fish exhibited interspecific and intraspecific variations, resulting from the differences in lipid content, sex, and body sizes (length and mass) between or within species. Both p,p'-DDMU and p,p'-DDM were consistently detected in the fish eggs, demonstrating their maternal transfer in female fish. The mean eggs to liver lipid-normalized concentration (E/L) ratios of p,p'-DDMU and p,p'-DDM were 0.98 and 1.77 in the northern snakehead, 0.35 and 0.01 in crucian carp, respectively; which were comparable to or even exceeded those of DDT and its major metabolites calculated in the same individual. Statistical analyses of the data showed that the E/L ratios were positively correlated with body sizes of the fish, but negatively correlated with the hepatic concentrations of p,p'-DDMU and p,p'-DDM in females; suggesting the influences of fish sizes and the mother body residues on their maternal transfer efficiencies.

Halogenated flame retardants in wild, prey-sized mud carp from an e-waste recycling site in South China, 2006-2016: Residue dynamics and ecological risk assessment.

The crude e-waste recycling has been regulated in China since the late 2000s; however, information on the recent levels and the ecological risks of e-waste derived contaminants such as halogenated flame retardants (HFRs) in the e-waste sites are limited. We therefore examined the concentrations of several HFRs in wild, prey-sized mud carps collected from a typical e-waste site in 2006, 2011 and 2016, to understand the exposure dynamics and ecological risk of these chemicals. Several ecological and biological parameters including δ15N, δ13C, body size and lipid content of the fish were also examined, to ensure an overall uniformity of the sample set among the sampling years. Among the HFRs measured, polybrominated diphenyl ethers (PBDEs) were detected at the highest concentrations (contributing >90% to ∑HFRs), followed by Dechlorane Plus (DPs), polybrominated biphenyls (PBBs), and alternative brominated flame retardants (ABFRs). The fish concentrations of ∑PBDEs, ∑PBBs and ∑DPs significantly dropped by 65%, 57% and 53% from 2006 to 2011, and 12%, 74% and 51% from 2011 to 2016, respectively; likely reflecting the positive impact of the environmental regulations on crude e-waste recycling. The ∑ABFRs concentrations were also decreased by 80% from 2006 to 2011, but increased by 127% from 2011 to 2016; suggesting possible fresh input of these novel HFRs in recent years. In addition to the changes in the HFR concentrations, contaminant profiles in the fish were also changed, possibly due to environmental degradation of the HFRs. Despite our conservative method of risk assessment, we found that PBDEs posed an important risk both for the mud carp and for piscivorous wildlife that inhabit the e-waste site.

Role of delayed wider endoscopic optic decompression for traumatic optic neuropathy: a single-center surgical experience.

BACKGROUND: The aim of the present study was to discuss the efficacy of delayed wider endoscopic optic decompression in traumatic optic neuropathy (TON). METHODS: A total of 479 patients were treated with corticosteroids and delayed wider endoscopic optic decompression, including the injury-to-surgery interval, within 2 weeks in patients with no light perception (NLP), and within 1 month in patients with residual eyesight. Based on the traditional decompression range, the superior wall of the optic canal was further decompressed. The preoperative and postoperative visual acuities (VAs) were reviewed, and the therapeutic efficacy was analyzed. RESULTS: The final VA was 0.1 or better in 29 cases, finger count in 79 cases, hand motion in 99 cases, light perception (LP) in 25 cases, and NLP in 247 cases. A total of 136 patients (136/383, 35.5%) recovered after NLP treatment, and 78 patients (69/96, 71.9%) had improved residual eyesight. The improvement rate in patients with residual eyesight was significantly higher than that of patients with NLP (P<0.01). Moreover, the total VA after treatment was better than that before surgery (P<0.01). CONCLUSIONS: Delayed wider optic nerve decompression plus corticosteroids remains an effective and safe therapeutic strategy for patients with delayed treatment intervals of more than 1 week, especially for those with residual eyesight within 1 month.

Overexpression of CD155 is associated with PD-1 and PD-L1 expression on immune cells, rather than tumor cells in the breast cancer microenvironment.

BACKGROUND: CD155 is an immune checkpoint protein in cancers and interacts with ligands to regulate the immune microenvironment. The expression of CD155 is correlated with the prognosis and pathological features of breast cancer. AIM: To investigate the expression status of CD155 and the association with exhausted CD4+ helper and CD8+ cytotoxic tumor infiltrating lymphocytes (TILs) and PD-L1 in the breast cancer microenvironment. METHODS: One hundred and twenty-six breast cancer patients with invasive ductal breast cancer were consecutively recruited into this study. Immunohistochemistry was used to detect the expression CD155, PD-L1 and PD-1 on tumor-infiltrating immune cells and tumor cells in the microenvironment. RESULTS: The proportion of patients with CD155 expression was higher in triple negative breast cancer (72.7%) than in Luminal A patients (22.2%, P < 0.05). Patients with positive CD155 expression had a higher percentage of CD4+/PD-1+ helper TILs (30%) than patients with negative CD155 expression (21%, P < 0.05). Patients with positive CD155 expression also had higher cell counts of exhausted CD4+ TILs [47 vs 20/high-power fields (HPF)] and unexhausted CD8+ TILs (30 vs 17/HPF) than patients with negative expression (P < 0.05). CD155 expression was correlated with increased PD-L1 expression in immune cells, 0.8% and 0.02% immune cells expressed PD-L1 in patients with positive and negative CD155 expression, respectively (P < 0.05). CONCLUSION: CD155 was related to an inhibitory immune breast cancer microenvironment. CD155 was associated with a high proportion of exhausted CD4+ and unexhausted CD8+ TILs and high PD-L1 expression in immune cells.

Overexpression of an Immune Checkpoint (CD155) in Breast Cancer Associated with Prognostic Significance and Exhausted Tumor-Infiltrating Lymphocytes: A Cohort Study.

PURPOSE: The immune checkpoint inhibitor is approved for breast cancer treatment, but the low expression of PD-L1 limits the immunotherapy. CD155 is another immune checkpoint protein in cancers and interacts with ligands to regulate immune microenvironment. This study is aimed at investigating the expression of CD155 and the association with prognosis and pathological features of breast cancer. METHODS: 126 patients were recruited this cohort study consecutively, and CD155 expression on tumor cells was detected by immunohistochemistry. The Kaplan-Meier survival curve and Cox hazard regression model were used to estimate the association. RESULTS: 38.1% patients had an overexpression of CD155, and the proportion of tumor cells with CD155 overexpression was 17%, 39%, 37%, and 62% among Luminal A, Luminal B, HER2-positive, and triple negative breast cancer cases, respectively (p < 0.05). Patients with CD155 overexpression had the Ki-67 index significantly higher than that of patients with low expression (42% vs. 26%). Though the number of tumor-infiltrating lymphocytes was higher among patients with CD155 overexpression (144/HPF vs. 95/HPF), the number of PD-1+ lymphocytes was significantly higher (52/HPF vs. 25/HPF, p < 0.05). Patients of CD155 overexpression had the disease-free and overall survival decreased by 13 months and 9 months, respectively (p < 0.05). CD155 overexpression was associated with an increased relapse (HR = 13.93, 95% CI 2.82, 68.91) and death risk for breast cancer patients (HR = 5.47, 1.42, 20.99). CONCLUSIONS: Overexpression of CD155 was correlated with more proliferative cancer cells and a dysfunctional immune microenvironment. CD155 overexpression introduced a worse relapse-free and overall survival and might be a potential immunotherapy target for breast cancer.