Morphology, genetic characterization and phylogeny of Aplectana dayaoshanensis n. sp. (Nematoda: Ascaridida) from frogs.

Cosmocercoid nematodes are common parasites in the digestive tract of amphibians. However, our knowledge of the species diversity, genetic data and molecular phylogeny of the superfamily Cosmocercoidea are far from being well understood. In the present study, large numbers of cosmocercoid nematodes were collected from the fine-spined frog Sylvirana spinulosa (Smith) (Anura: Ranidae) and the white-spotted thigh tree-frog Polypedates megacephalus (Hallowell) (Anura: Rhacophoridae) in Guangxi Province, China. Integrated morphological and genetic evidence reveals these nematode specimens to be a new species of the genus Aplectana, A. dayaoshanensis n. sp. (Cosmocercoidea: Cosmocercidae). The molecular characterization of small ribosomal DNA (18S), internal transcribed spacer (ITS) and large ribosomal DNA (28S) of A. dayaoshanensis n. sp., together with the 28S of A. chamaeleonis (Baylis, 1929) (collected from Hyperolius kivuensis Ahl in Rwanda), were reported for the first time. Moreover, phylogenetic analyses using maximum likelihood (ML) inference based on 18S + 28S and ITS sequence data, respectively, both supported the family Cosmocercidae to be a monophyletic group and the family Kathlaniidae to be a paraphyletic group. Our phylogenetic results rejected the monophyly of the genus Aplectana. The present results contribute to the knowledge of the species diversity and genetic data of cosmocercoid nematodes, and preliminarily revealed the phylogenetic relationships of the major families and some genera in the Cosmocercoidea.

LncRNA HCG18 suppresses CD8+ T cells to confer resistance to cetuximab in colorectal cancer via miR-20b-5p/PD-L1 axis.

Aim: We aimed to explore the effect of long noncoding RNA HCG18 in colorectal cancer (CRC). Materials & methods: Relative gene and protein expression were screened. Colony formation and flow cytometry assays were performed to determine proliferation and apoptosis. Dual luciferase and RNA immunoprecipitation assays were conducted to validate the interaction between indicated molecules. Xenograft in nude mice was applied to verify the conclusion in vivo. Results:HCG18 and PD-L1 were upregulated while miR-20b-5p was downregulated in CRC tissue. Functional analysis revealed that lncRNA HCG18 promoted proliferation, migration and resistance to cetuximab of CRC cells via the miR-20b-5p/PD-L1 axis. Conclusion:HCG18 facilitated progress of the tumor, conferred to cetuximab resistance and suppressed CD8+ T cells via the miR-20b-5p/PD-L1 axis.

Lay abstract In the present study, we found a long noncoding RNA (lncRNA), HCG18 (a recently discovered lncRNA that facilitates tumor progression via multiple mechanisms), was upregulated in colorectal cancer (CRC). Further studies revealed that HCG18 suppressed CD8⁺ T-cell (cytotoxic T lymphocyte which kills cancer cell) activation to induce cetuximab (a first-line drug in CRC) resistance. Mechanically, HCG18 elevated expression of PD-L1 (a receptor in T-cell membranes, thus suppressing the proliferation of CD8⁺ cytotoxic T lymphocytes) via sponging (lncRNA binds with miRNA) miR-20b-5p. This study might provide a deeper insight into understanding cetuximab resistance in CRC.

Hsa_circ_0136666 activates Treg-mediated immune escape of colorectal cancer via miR-497/PD-L1 pathway.

PURPOSE: In the rankings of cancer mortality and incidence worldwide, colorectal cancer ranks fourth and the third, respectively. Circular RNA hsa_circ_0136666 (hsa_circ_0136666) is reported to participate in the growth of colorectal cancer. However, the mechanism by which hsa_circ_0136666 regulates the tumorigenesis of colorectal cancer needs to be further explored. In this study, we report here the role of hsa_circ_0136666 in the aberrant activation of Treg cells and immune evasion of tumor cells, providing a new strategy for the treatment of colorectal cancer. METHODS: Western blotting assay and qRT-PCR assay were used to determine protein and mRNA expression levels. Dual-luciferase reporter assay was used to evaluate the targeted regulatory relationship. RNA immunoprecipitation was used to detect RNA binding. Colony formation assay was utilized to measure the cell proliferation. Flow cytometry was used to assess cell apoptosis. Xenograft model was setup to evaluate tumor growth. RESULTS: The results showed that hsa_circ_0136666 and PD-L1 was increased in colorectal cancer cells while miR-497 was decreased in colorectal cancer cells when compared with normal colon epithelial cell line. Hsa_circ_0136666 was demonstrated to directly target miR-497, which also regulated PD-L1 by binding to its 3'UTR. Further mechanistic studies identified that hsa_circ_0136666 controlled cell proliferation and apoptosis via targeting miR-497 and regulating PD-L1 expression. Of note, hsa_circ_0136666 stimulated Treg cells mediated by miR-497/PD-L1 axis and its downstream signal pathway in Treg cells. Finally, hsa_circ_0136666 was found to accelerate the tumor growth in vivo. CONCLUSIONS: Our findings demonstrated that hsa_circ_0136666 promoted the expression of PD-L1 by inhibiting miR-497 level in colorectal cancer, thus inducing the activation of Treg cells and leading to the immune escape of tumor, providing a novel mechanistic insight into the pathogenesis of colorectal cancer.

Description of a new species of Aplectana (Nematoda: Ascaridomorpha: Cosmocercidae) using an integrative approach and preliminary phylogenetic study of Cosmocercidae and related taxa.

BACKGROUND: Nematodes of the family Cosmocercidae (Ascaridomorpha: Cosmocercoidea) are mainly parasitic in the digestive tract of various amphibians and reptiles worldwide. However, our knowledge of the molecular phylogeny of the Cosmocercidae is still far from comprehensive. The phylogenetic relationships between Cosmocercidae and the other two families, Atractidae and Kathlaniidae, in the superfamily Cosmocercoidea are still under debate. Moreover, the systematic position of some genera within Cosmocercidae remains unclear. METHODS: Nematodes collected from Polypedates megacephalus (Hallowell) (Anura: Rhacophoridae) were identified using morphological (light and scanning electron microscopy) and molecular methods [sequencing the small ribosomal DNA (18S), internal transcribed spacer 1 (ITS-1), large ribosomal DNA (28S) and mitochondrial cytochrome c oxidase subunit 1 (cox1) target regions]. Phylogenetic analyses of cosmocercoid nematodes using 18S + 28S sequence data were performed to clarify the phylogenetic relationships of the Cosmocercidae, Atractidae and Kathlaniidae in the Cosmocercoidea and the systematic position of the genus Aplectana in Cosmocercidae. RESULTS: Morphological and genetic evidence supported the hypothesis that the nematode specimens collected from P. megacephalus represent a new species of Aplectana (Cosmocercoidea: Cosmocercidae). Our phylogenetic results revealed that the Cosmocercidae is a monophyletic group, but not the basal group in Cosmocercoidea as in the traditional classification. The Kathlaniidae is a paraphyletic group because the subfamily Cruziinae within Kathlaniidae (including only the genus Cruzia) formed a seperate lineage. Phylogenetic analyses also showed that the genus Aplectana has a closer relationship to the genus Cosmocerca in Cosmocercidae. CONCLUSIONS: Our phylogenetic results suggested that the subfamily Cruziinae should be moved from the hitherto-defined family Kathlaniidae and elevated as a separate family, and the genus Cosmocerca is closely related to the genus Aplectana in the family Cosmocercidae. The present study provided a basic molecular phylogenetic framework for the superfamily Cosmocercoidea based on 18S + 28S sequence data for the first time to our knowledge. Moreover, a new species, A. xishuangbannaensis n. sp., was described using integrative approach.

The hepatic microenvironment promotes lung adenocarcinoma cell proliferation, metastasis, and epithelial-mesenchymal transition via METTL3-mediated N6-methyladenosine modification of YAP1.

The inflammatory microenvironment plays an important role in the onset and progression of lung adenocarcinoma (LUAD), and the liver is a suitable site of metastasis for LUAD cells. However, whether the inflammatory microenvironment of the liver is conducive to the proliferation, invasion, and metastasis of LUAD cells remains unclear. In this study, we confirmed that the hepatic inflammatory microenvironment stimulated by IL-6 promoted the proliferation, migration, invasion, and epithelial-mesenchymal transition of LUAD cells, increased the m6A methylation of total RNA, and transcriptionally activated METTL3 expression. Additionally, METTL3 activated the YAP1/TEAD signaling pathway by increasing the m6A modification and expression of YAP1 mRNA. These results indicate that the hepatic inflammatory microenvironment plays a role in regulating the biological functions of LUAD cells. Further, our study identifies a molecular mechanism that may provide a new strategy for the early diagnosis, treatment, and prognosis of liver metastasis in LUAD patients.

Effect of C-reactive protein/albumin ratio on prognosis in advanced non-small-cell lung cancer.

AIM: Systemic inflammatory response is closely related to tumor progression. We retrospectively investigated relationships between systemic inflammatory scores, C-reactive protein/albumin (CRP/Alb) ratio (CAR) and clinical characteristics in advanced non-small-cell lung cancer (NSCLC) in 436 patients to find better clinical predictors of NSCLC prognosis. METHODS: Blood specimens were collected 1 week before treatment to test for systemic inflammatory scores and albumin. Patients' overall survival (OS) was calculated via Kaplan-Meier method. Single-factor log-rank and multivariate Cox regression analyses and receiver operating characteristic curves were used to evaluate the prognostic significance of CAR and other systemic inflammatory indexes in predicting OS. RESULTS: Kaplan-Meier method showed that Glasgow prognosis score (GPS), modified GPS (mGPS), neutrophil/lymphocyte ratio (NLR), platelet/lymphocyte ratio (PLR) and monocyte/lymphocyte ratio (MLR) were reliable prognostic factors for advanced NSCLC. CAR was positively correlated with GPS, mGPS, NLR, PLR and MLR in these patients. CAR was an independent risk factor for OS in advanced NSCLC, and was more closely associated with prognosis than were GPS, mGPS, NLR, PLR or MLR. CONCLUSION: In advanced NSCLC patients, CAR may be a better predictor of prognosis compared with other inflammatory markers. A prospective multicenter study is needed to verify these findings.

Recurrent Proliferative Glomerulonephritis With Monoclonal IgG Deposits After a Renal Transplant Which Was Insensitive to Pulse Therapy Remitted by Double Filtration Plasmapheresis.

Proliferative glomerulonephritis with monoclonal IgG deposits manifesting as a nephrotic syndrome recently has been described as a renal disease with the pathological features of mesangial and subendothelial deposits of monoclonal IgG. Eight cases of recurrent proliferative glomerulonephritis with monoclonal IgG deposits after a renal transplant have been reported. Almost all of these patients had a certain remission of proteinuria by steroids alone or with cyclophosphamide, and had further remission through other special treatments (ie, rituximab and plasmapheresis). We present a case of recurrent proliferative glomerulonephritis with monoclonal IgG deposits of the IgG3? subtype after a renal transplant, which was insensitive to pulse intravenous methyl-prednisolone and cyclophosphamide remitted by double filtration plasmapheresis. This case report reveals that recurrent proliferative glomerulo-nephritis with monoclonal IgG deposits may be insensitive to intravenous pulse therapy of methylprednisolone and cyclophosphamide. We advocate double filtration plasmapheresis as an effective treatment of proliferative glomerulo-nephritis with monoclonal IgG deposits on remission of proteinuria.

Elevated serum C-reactive protein, carcinoembryonic antigen and N2 disease are poor prognostic indicators in non-small cell lung cancer.

AIM: To assess the prognostic value of mediastinal lymph node metastases (N2 disease), carcinoembryonic antigen (CEA) levels and C-reactive protein (CRP) in non-small cell lung cancer (NSCLC), according to the 7th edition of the TNM classification. METHODS: Newly diagnosed stage III-IV NSCLC were enrolled, including 75 patients with malignant pleural effusion. The relationship between serum CRP levels and other relevant variables such as sex, Eastern Cooperative Oncology Group status, smoking status, initial staging, N2 disease, serum albumin, white blood cell count, platelet count, CEA, comorbidity and pathology were analyzed. Univariate and multivariate analyses were performed to find prognostic markers using Cox's proportional hazards model. RESULTS: Of the 127 patients enrolled, 55 (43%) had elevated CRP levels. There was a significant correlation between serum CRP level and platelet count (P = 0.011). Median overall survival (OS) in the normal CRP group was significantly longer than in the high CRP group (15.7 months vs 9.1 months, P = 0.013). Hypoalbuminemia (P = 0.047), higher CEA (P = 0.043) and N2 disease (P = 0.040) were additional prognostic factors on univariate analysis. On multivariate analysis an elevated CRP serum level (HR = 1.796; P = 0.005), higher CEA (HR = 1.563; P = 0.031) and N2 disease (HR = 1.723; P = 0.012) were independent prognostic factors for poor survival. CONCLUSION: High levels of serum CRP and CEA, and N2 disease are independent prognostic indicators for the survival of patients with stage III-IV NSCLC.

Serum VEGFR-3 and survival of advanced gastric cancer patients treated with FOLFOX.

AIM: To explore if vascular endothelial growth factor receptor-3 (VEGFR-3) and carcinoembryonic antigen (CEA) can predict overall survival in advanced gastric cancer. METHODS: VEGFR-3 level was assessed by enzyme-linked immunosorbent assay, and CEA was assessed by chemiluminescence immunoassay in the sera of 81 advanced gastric cancer patients before treatment with oxaliplatin plus 5-fluorouracil and folinic acid. RESULTS: Median survival time in patients with a low serum VEGFR-3 level was significantly longer than in those with a higher VEGFR-3 level (15.4 mo vs 7.7 mo, P < 0.001). Patients with a low CEA level had a longer survival than those with a higher CEA level (15.8 mo vs 8.6 mo, P < 0.001). Thirty-nine patients with low VEGFR-3 and low CEA levels had a median survival of 19.7 mo (P = 0.0006). The hazard ratio for patients with a high VEGFR-3 level was 2.443 (P = 0.002). CONCLUSION: High serum VEGFR-3 level is correlated significantly with poor survival. In patients with a high serum level of VEGFR-3, alternative chemotherapy regimens should be considered.