Retraction Note: Study on the functions and mechanism of immune functions of human telomerase reverse transcriptase regulating dendritic cells treating sepsis.

The article "Study on the functions and mechanism of immune functions of human telomerase reverse transcriptase regulating dendritic cells treating sepsis", by H.-M. Chen, L.-Q. Wang, H.-P. Wan, H.-Z. Wei, L.-C. Ke, C.-Y. Liu, Q.-Y. Tan, published in Eur Rev Med Pharmacol Sci 2016; 20 (21): 4500-4507-PMID: 27874963 has been retracted by the Editor in Chief for the following reasons. Some concerns were raised on PubPeer (https://pubpeer.com/publications/3604386A706802443E51758A893D6F) about Figures 3, 4, and 5 showing some overlaps and similar bands in Western blots figures. Furthermore, there is a lack of information regarding the ethics approval for the study involving rats. The journal contacted the authors to request the original raw data and information regarding the ethical approval of the manuscript but never received a reply. Therefore, due to major concerns detected, the Editor in Chief mistrusts the results presented and decided to withdraw the manuscript. The corresponding author has been informed about the retraction. This article has been retracted. The Publisher apologizes for any inconvenience this may cause. https://www.europeanreview.org/article/11476.

LncRNA WT-AS inhibits metastatic ability of non-small cell lung cancer by regulating KLK13.

OBJECTIVE: The aim of this study was to investigate the effect of long non-coding RNA (lncRNA) WT-AS on the invasiveness and migration of non-small cell lung cancer (NSCLC) cells, and to explore the underlying molecular mechanism of lncRNA WT-AS in the pathogenesis of NSCLC. PATIENTS AND METHODS: LncRNA WT-AS expression in 50 pairs of NSCLC tissues and adjacent ones was studied by quantitative real-time polymerase chain reaction (qRT-PCR) analysis, and the correlations of WT-AS with clinicopathological indicators and prognosis of NSCLC patients were analyzed. Meanwhile, NSCLC expression levels in NSCLC cell lines were also evaluated by qPCR assay. In addition, WT-AS overexpression and knockdown models were constructed using lentivirus in NSCLC cell lines A549 and H1299, respectively. Thereafter, transwell and cell wound healing assays were carried out to assess the implication of WT-AS in biological functions of NSCLC cells. Furthermore, the interaction between WT-AS and KLK13 was determined via Luciferase assay. RESULTS: The results showed that WT-AS expression in NSCLC was remarkably lower than that in normal tissues adjacent to the cancer. Univariate analysis suggested that compared with patients with high expression of WT-AS, patients in low expression group showed higher incidence of metastasis and lower survival rates. Overexpression of WT-AS suppressed cell invasion and metastasis capacity, while the opposite result was observed in WT-AS knockdown group. KLK13 expression showed an increase in NSCLC cell lines and tissues, which was negatively correlated with WT-AS level. Meanwhile, Luciferase assay confirmed the binding between WT-AS and KLK13. Western blotting revealed that KLK13 expression was remarkably elevated in EC tissues and was positively correlated with TRIM62. In addition, it was also found that WT-AS and KLK13 had a mutual regulatory effect, which together affect the malignant progress of NSCLC. CONCLUSIONS: This study shows for the first time that LncRNA WT-AS interacts with KLK13 to serve as a negative regulator of NSCLC progression.

Upregulation of caspase-3 by high glucose in chondrocyte involves the cytoskeleton aggregation.

OBJECTIVE: The hyperglycemic environment of diabetes promotes chondrocyte (CH) apoptosis and is closely related to the occurrence and development of osteoarthritis (OA). This present study aimed to elucidate the relation between the cytoskeleton and the caspase-3 expression of human CHs in high glucose in vitro. PATIENTS AND METHODS: We used different concentrations of glucose medium to test the effect of glucose on the CHs viability. Cytochalasin D and colchicine were used to prevent the aggregation of F-actin and ß-tubulin. Besides, Z-DEVD-FMK (ZDF) or Apoptosis Activator 2 was used to inhibit or activate the caspase-3 expression. The intensity of F-actin and ß-tubulin, cell viability, apoptosis, and caspase-3 expression were analyzed. RESULTS: Three days of treatment of 30 mM or 40 mM glucose significantly decreased the CHs viability compared to the 10 mM but increased the caspase-3, apoptosis, collagen, and the aggregation of the F-actin and ß-tubulin. However, the cytochalasin D and colchicine partly rejected the high-glucose induced caspase-3 upregulation, apoptosis, and CHs disability. Besides, these two anti-aggregation drugs also suppressed the Apoptosis Activator 2 induced caspase-3 upregulation and apoptosis. Furthermore, the application of ZDF could only prevent the F-actin aggregation, but not the ß-tubulin. CONCLUSIONS: Long-term high glucose triggers the caspase-3 expression and leads to the CH apoptosis involving cytoskeleton aggregation. Inhibition of cytoskeleton aggregation through the F-actin or ß-tubulin could alleviate the high glucose-induced caspase-3 upregulation.

Investigating the expression of miRNA-133 in animal models of myocardial infarction and its effect on cardiac function.

OBJECTIVE: This study aimed to investigate the expression of miR-133 in animal models of myocardial infarction and its effect on cardiac function. MATERIALS AND METHODS: Forty-five male-specific pathogen-free (SPF) C57/BL6 mice were selected, among which 35 were made for animal models of myocardial infarction and were enrolled into Model Group and another 10 were enrolled into Blank Control Group. Seven mice died in the model making. Ten mice randomly selected from the 28 mice successfully modeled were transfected with adenovirus carrying miRNA-133 and set as Virus Group, while the remaining 18 mice were randomly divided into Virus No-load Group and Model Group. Mice in Virus Group were transfected with adenovirus carrying miRNA-133, while those in Virus No-load Group were transfected with empty viral vectors without miRNA-133. The left ventricular ejection fraction (LVEF) and fractional shortening (FS) of the mice weeks after the infection were recorded and evaluated by echocardiography. The relative expression levels of miR-133 in the heart tissues of the four groups of mice were compared by Real Time-Polymerase Chain Reaction (RT-PCR). RESULTS: The miR-133 expressions in Blank Control Group and Virus Group were higher than that of Model Group (p<0.05). Then, the myocardial infarction area of mice was compared. The LVEF and FS values of mice in Model Group, Virus No-load Group, and Virus Group were significantly lower than those in Blank Control Group, with the LVEF and FS values of Virus Group higher than that of Model Group and Virus No-load Group (p<0.05). The swimming time of Blank Control Group was significantly higher than that of Model Group and Virus Group (p<0.05), with Virus Group and Virus No-load Group having a greatly longer swimming time than Model Group (p<0.05). The myocardial infarction area of mice in Virus Group was significantly smaller than that in Model Group and Virus No-load Group, the difference was statistically significant (p<0.05). There was no significant difference in myocardial infarction area of mice between Model Group and Virus No-load Group (p>0.05). CONCLUSIONS: MiR-133 was in a low expression state in the mice models of myocardial infarction and the overexpression of miR-133 could significantly improve the cardiac function index and motor function, as well as reduce the myocardial infarction area of mice with myocardial infarction. This could inspire new molecular therapy for the treatment of myocardial infarction.

MiR-96-5p promotes the proliferation, invasion and metastasis of papillary thyroid carcinoma through down-regulating CCDC67.

OBJECTIVE: MicroRNAs (miRNAs) have been identified to play an important regulatory role in various biological behaviors of papillary thyroid carcinoma (PTC). However, the specific role and function of miR-96-5p in PTC remain unclear. Therefore, the aim of this study is to detect the expression of miR-96-5p in PTC, and to explore its exact function. PATIENTS AND METHODS: The relative expression level of miR-96-5p in PTC tissues and cell lines was detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). MiR-96-5p mimics or inhibitors were then constructed and transfected into cells to upregulate or downregulate miR-96-5p expression. MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assay and colony formation assay were employed to evaluate the proliferation of PTC cells. Meanwhile, transwell assay was employed to detect the invasion and metastasis of PTC cells. In addition, the underlying mechanism of miR-96-5p was identified by Luciferase reporter gene assay and Western blot analysis. RESULTS: The expression of miR-96-5p in PTC tissues and PTC-derived cell lines was significantly higher than that of normal controls. The overexpression of miR-96-5p remarkably promoted the proliferation, invasion and migration of PTC cells. However, knockdown of miR-96-5p significantly decreased cell growth and metastasis. CCDC67 was verified as a target gene of miR-96-5p in PTC. Further experiments demonstrated that the restoration of CCDC67 could significantly reduce the carcinogenic function of miR-96-5p. CONCLUSIONS: MiR-96-5p was remarkably upregulated in PTC tumor tissues and cells. In addition, it promoted cell growth, invasion, and migration via repressing CCDC67 expression.

Downregulation of plasma miR-124 expression is a predictive biomarker for prognosis of glioma.

OBJECTIVE: MicroRNAs (miRNAs) have been widely identified as potential biomarkers for predicting prognosis of glioma. The objective of the study was to examine the clinical role of plasma miR-124 expression in glioma. PATIENTS AND METHODS: MiR-124 expression in plasma samples from 64 cases glioma patients and 40 normal healthy controls was examined by quantitative reverse transcription PCR (qRT-PCR). The correlation of miR-124 expression with clinicopathological feathers or prognosis of glioma patients was assessed. Univariate and multivariate Cox analysis were used to analyze the risk factors of prognosis. The receiver operating characteristic (ROC) curve was established, and the area under the ROC curve (AUC) was calculated to evaluate the difference of miR-124 expression in glioma patients. RESULTS: We showed that miR-124 expression was significantly downregulated in plasma samples of glioma patients compared with normal healthy controls (p<0.05). Plasma miR-124 expression significantly associated with Karnofsky Performance Status (KPS) score (p<0.05) and WHO grade (p<0.05) in glioma plasma. Patients with low miR-124 expression had worse disease free survival (DFS) and overall survival (OS) time than patients with high miR-124 expression. Univariate and multivariate analysis implied that low miR-124 expression was an independent maker of disease free survival (DFS) and overall survival (OS) in glioma. CONCLUSIONS: Our findings implied that plasma miR-124 expression may serve as an independent predictor of poor prognosis.

The expression research of miR-210 in the elderly patients with COPD combined with ischemic stroke.

OBJECTIVE: We conduct this study to investigate the expression level of miR-210 in elderly patients with COPD combined with ischemic stroke and analyzed its application value as a sensitive diagnostic indicator. PATIENTS AND METHODS: 50 cases of elderly patients with COPD combined with ischemic stroke were selected as group A, 50 cases of elderly patients with COPD were selected as group B, 50 cases of elderly patients with ischemic stroke were selected as group C, and 50 cases of healthy volunteers as group D. Real-time PCR assay for quantification was used to detect the expression level of miR-210 in peripheral blood and receiver operating curve (ROC) was used to analyze the diagnostic sensitivity and accuracy. RESULTS: MiR-210 level of group A was the lowest, followed by group B and group C; group D had the highest levels. The difference was statistically significant (p<0.05). MiR-210 levels decreased with an increasing decline degree of lung function and the difference was statistically significant (p<0.05). After miR-210 diagnosis, COPD sensitivity was 85.6%, the specificity was 72.6%, accuracy (AUC) was 0.821, and 95% CI 0.632-0.924. CONCLUSIONS: The down-regulation of MiR-210 expression in the COPD combined with ischemic stroke can be regarded as a sensitive index in diagnosis of COPD and ischemic stroke.

Exosomes mediated transfer of lncRNA UCA1 results in increased tamoxifen resistance in breast cancer cells.

OBJECTIVE: In this study, we firstly compared the loading of urothelial carcinoma-associated 1 (UCA1) in exosomes between tamoxifen sensitive and tamoxifen resistant breast cancer cells and further investigated the role of exosomal transfer of UCA1 in the development of tamoxifen resistance in estrogen receptor (ER) positive breast cancer cells. MATERIALS AND METHODS: Exosomes were isolated from the culture medium of tamoxifen sensitive MCF-7 cells and tamoxifen resistant LCC2 cells. QRT-PCR was performed to analyze UCA1 expression in cells and exosomes. CCK-8 assay, immunofluorescence staining of cleaved caspase-3 and flow cytometric analysis of annexin V/PI staining were used to assess tamoxifen sensitivity. RESULTS: UCA1 is significantly increased not only in LCC2 cells, but also in exosomes released from LCC2 cells. The increase in exosomes is more evident than in cells. MCF-7 cells pretreated with exos/LCC2 had a significantly increased cell viability, a decreased expression of cleaved caspase-3 and a lower ratio of apoptosis after tamoxifen treatment. The exos/LCC2 with impaired UCA1 loading had significantly suppressed capability to promote tamoxifen resistance in MCF-7 cells. CONCLUSIONS: UCA1 is significantly loaded in exosomes from tamoxifen resistant LCC2 cells. Exosomes mediated transfer of UCA1 can significantly increase tamoxifen resistance in ER-positive MCF-7 cells.

Study on the functions and mechanism of immune functions of human telomerase reverse transcriptase regulating dendritic cells treating sepsis.

OBJECTIVE: We analyzed the functions and mechanisms of immune functions of human telomerase reverse transcriptase regulating dendritic cells (DC) treating sepsis of mice models. MATERIALS AND METHODS: Eighty clean grade Balb/c animals aged from 6 to 8 weeks, weighted from 18 g to 22 g were selected for this study. The DC cells were harvested from the animals and cultivated to transfect with the recombinant eukaryotic expression plasmid hTERT-IRES2-EGFP construct. The LPS (E. coli 0111:B4, 5 mg/kg) was injected into the abdominal cavity of mice to establish sepsis models. Afterwards, animals were divided randomly into the sepsis group (A group), the group of hTERT transfecting DC (B group), the group of DC un-transfected (C group) with 25 mice in each group. 5 mice were in the normal control group (D group), without any treatment. An equivalent volume of normal saline was injected into the abdominal cavity of A group. Subsequently, 1 ml of cell suspension (105/ml) was transfected into B and C groups respectively. Five animals from A, B, C groups and one animal from group D were sacrificed after 24h, 48h, 72d, 7d and 10d respectively. RESULTS: It was found that median survival time of the group of hTERT transfecting DC was remarkably higher than that of the untransfected group and the sepsis group. The average scores of the pathology of kidney and intestine at each time were significantly lower than that of the other two groups (p<0.05). At each time point, in the group of hTERT transfecting DC, levels of CRP and Cr were remarkably lower than that of the other two groups; HLA-DR, CD40 of immune phenotype and the expression level of peripheral blood T cells MHC-II molecules were significantly higher than that of the other two groups; the expression level of IL-12 and TNF-a were significantly lower than that of the other two groups; apoptosis rate of DC were significantly lower than that of the other two groups; the content and activity of NF-κB were significantly higher than that of the other two groups (p<0.05). CONCLUSIONS: The telomerase reverse transcriptase gene can raise the expression and maturity of DC, reduce apoptosis, induce cytokine secretion, reduce the inflammatory response and increase the survival time.

Specificity redirection by CAR with human VEGFR-1 affinity endows T lymphocytes with tumor-killing ability and anti-angiogenic potency.

Immunotherapy that is based on adoptive transfer of T lymphocytes, which are genetically modified to express chimeric antigen receptors (CARs) that recognize tumor-associated antigens, has been demonstrated to be an efficient cancer therapy. Vascular endothelial growth factor receptor-1 (VEGFR-1), a vital molecule involved in tumor growth and angiogenesis, has not been targeted by CAR-modified T lymphocytes. In this study, we generated CAR-modified T lymphocytes with human VEGFR-1 specificity (V-1 CAR) by electroporation. V-1 CAR-modified T lymphocytes were demonstrated to elicit lytic cytotoxicity to target cells in a VEGFR-1-dependent manner. The adoptive transfer of V-1 CAR T lymphocytes delayed tumor growth and formation and inhibited pulmonary metastasis in xenograft models and such efficacies were enhanced by cotransfer of T lymphocytes that expressed interleukin-15 (IL-15). Moreover, V-1 CAR-modified T lymphocytes lysed primary endothelial cells and impaired tube formation, in vitro. These data demonstrated the antitumor and anti-angiogenesis ability of V-1 CAR-modified T lymphocytes. Our study provides the rationale for the clinical translation of CAR-modified T lymphocytes with VEGFR-1 specificity.

Combination of targeting gene-viro therapy with recombinant Fowl-pox viruses with HN and VP3 genes on mouse osteosarcoma.

BACKGROUND: Osteosarcoma is an aggressive cancerous neoplasm arising from primitive transformed cells of mesenchymal origin that exhibit osteoblastic differentiation and produce malignant osteoid. With the rapid development of tumor molecular biology, gene and viral therapy, a highly promising strategy for the treatment, has shown some therapeutic effects. OBJECTIVES: To study the strategy of cooperative cancer gene therapy, previously, we explored the antitumor effects of recombinant Fowl-pox viruses (FPVs) with both HN (hemagglutinin-neuramidinase) and VP3 genes on mouse osteosarcoma. MATERIALS AND METHODS: We constructed vFV-HN, vFV-VP3 and vFV-HN-VP3 inserting CAV VP3 gene, NDV HN gene into fowlpox virus. S180 osteosarcoma were transfected with Recombinant Fowl-pox viruses (FPVs). These cell lines stably expressing tagged proteins were selected by culturing in medium containing puromycin (2 µg/ml) and confirmed by immunoblotting and immunostaining. S180 osteosarcoma model with BALB/c mice and nude mice were established and the vFPV viruses as control, vFV-HN, vFV-VP3, vFV-HN-VP3 were injected into the tumor directly. The rate of tumor growth, tumor suppression and the sialic acid levels in serum were examined and the tumor tissues were analyzed by the method of immunohistochemistry. Flow cytometric analysis was performed using a FACSCalibur flow cytometer. A total of 100,000 events were analyzed for each sample and the experiment was repeated at least twice. RESULTS: Our data indicated that vFV-HN, vFV-VP3 and vFV-HN-VP3 all had growth inhibition effects, the inhibition rate of vFV-HN-VP3 group was 51.7%, which was higher than that of vFV-HN, vFV-VP3 group and control group (p < 0.01). The sialic acid level of vFV-HN-VP3 group in mouse serum was 4.22±0.27 mmol/l, which was lower than that of other groups (p < 0.01). CONCLUSIONS: These results suggest that genes into mouse osteosarcoma cancer cells can cause cell a specificity anti-tumor immune activity, suppress tumor growth, and increase the survival rate of the tumor within host.

Probing the geometry and interconnectivity of pores in organic aerogels using hyperpolarized 129XE NMR spectroscopy.

Hyperpolarized (HP) 129Xe NMR was used for the first time to probe the geometry and interconnectivity of pores in RF aerogels and to correlate them with the [resorcinol]/[catalyst] (R/C) ratio. We have demonstrated that HP 129Xe NMR is an ideal method for accurately measuring the volume-to-surface-area (Vg/S) ratios for soft mesoporous materials without using any geometric models. The Vg/S parameter, which is related both to the geometry and the interconnectivity of the porous space, has been found to correlate strongly with the R/C ratio and exhibits an unusually large span: an increase in the R/C ratio from 50 to 500 results in about 5-fold rise in Vg/S. Unlike conventional techniques, HP 129Xe NMR spectroscopy probes the geometry and interconnectivity of the nano- or mesopores in soft materials, providing new insights into the pore structure.

Grayanane diterpenoids from pieris formosa

Two new diterpenoids, pierisformoside A (1) and pierisformosin D (2), and two known diterpenoids, asebotoxins VIII (3) and V (4), were isolated from leaves of Pieris formosa. Their structures were elucidated on the basis of spectral analysis, including 1H-1H COSY, 13C-1H COSY, HMBC, and NOESY experiments.