Phosphorylation-mediated interaction between human E26 transcription factor 1 and specific protein 1 is required for tumor cell migration.

Transcription factors, human E26 transcription factor 1 (Ets1) and specific protein 1 (Sp1), are known to induce gene expression in tumorigenicity. High Ets1 expression is often associated with colorectal tumorigenesis. In this study, we discover that metastasis and clone formation in SW480 cells mainly depend on the direct interaction between Ets1 and Sp1 instead of high Ets1 expression. The interaction domains are further addressed to be the segment at Sp1(626-708) and the segment at Ets1(244-331). In addition, the phosphorylation inhibition of Ets1 at Tyr283 by either downregulation of Src kinase or Src family inhibitor treatment decreases the interaction between Sp1 and Ets1 and suppresses SW480 migration. Either administration or overexpression of the peptides harboring the interaction segment strongly inhibits the colony formation and migration of SW480 cells. Our findings suggest that the interaction between Ets1 and Sp1 rather than Ets1 alone promotes transformation in SW480 cells and provide new insight into the Ets1 and Sp1 interaction as an antitumour target in SW480 cells.

Multi-Omics Integration Reveals the Crucial Role of Fusobacterium in the Inflammatory Immune Microenvironment in Head and Neck Squamous Cell Carcinoma.

The tumor microbiome is believed to have a profound impact on tumor progression owing to its local colonization in the tumor microenvironment (TME). Using the Cancer Microbiome Atlas (TCMA), a database of curated, decontaminated microbial profiles for 3,689 oropharyngeal, esophageal, gastrointestinal, and colorectal tissue samples from 1,772 patients, we conducted a comprehensive multi-omics analysis to reveal microbial signatures among various cancers and the potential mechanisms involved in tumor progression of head and neck squamous cell carcinoma (HNSC). We found that compared with other cancer types, the tumor-resident microbiome of HNSC accounted for the highest bacterial abundance and strongest association with host TME signatures. Fusobacterium was found to be enriched in HNSC tissues, which was associated with an increased inflammatory effect and inferior prognosis. Moreover, we revealed that the microbiota-associated inflammatory TME was attributed to the competing endogenouse RNA (ceRNA) network and chromatin accessibility. IMPORTANCE Studies on revealing the composition and potential mechanisms of the tumor microbiome are still at an initial stage. We uncovered the potential contribution of the tumor-resident microbiota on the immunosuppressive microenvironment in HNSC, which will provide a new perspective for tumor microbiome research and yield valuable insights into the clinical management of HNSC.

CircRNA-011235 Counteracts The Deleterious Effect of Irradiation Treatment on Bone Mesenchymal Stem Cells by Regulating The miR-741-3p/CDK6 Pathway.

OBJECTIVE: The present work was aimed at uncovering the effect of circRNA-011235 (circ-011235) on irradiation-induced bone mesenchymal stem cells (BMSCs) injury and its regulatory mechanism, with a view to establish a scientific basis for its possible medical applications. MATERIALS AND METHODS: In this experimental study, after irradiation with different doses (0, 2, 4, 6 GY), the relative expression levels of circ-011235, miR-741-3p, and cyclin-dependent kinases 6 (CDK6) were detected in the BMSCs, using the real time-quantitative polymerase chain reaction (RT-qPCR). The overexpression effects of circ-011235 and CDK6 on the cell proliferation in irradiation-treated BMSCs were measured by the Cell Counting Kit-8 (CCK8) assay. And also, their effects on the cell cycle were evaluated by flow cytometry. RT-qPCR and immunoblotting were performed to detect the effects of pcDNA-circ-011235 and pcDNA-CDK6 on the expression of cyclin D1 and cyclindependent kinases 4 (CDK6) at the gene and protein levels, respectively. RESULTS: Irradiation treatment elevated the expression of circ-011235 and CDK6, but reduced miR-741-3p expression in the BMSCs with a dose-dependent effect. The proliferation of BMSCs was significantly inhibited in the irradiation treatment group, while the overexpression of circ-011235 and CDK6 effectively attenuated this inhibition. Also, overexpression of circ-011235 and CDK6 elevated the expression of cyclin D1 in irradiation-treated BMSCs, but had no significant effect on the CDK4 expression. CONCLUSION: Our results demonstrated that circ-011235 up-regulated the expression of cyclin D1 via miR-741-3p/ CDK6 signal pathway, thereby promoting cell cycle progression and proliferation of irradiation-treated BMSCs. This finding suggested circ-011235/ miR-741-3p/CDK6 pathway exerted a protective role in the response to irradiation and will be a potential new target for future research on the mechanism involved in the resistance of BMSCs to radiation.

CircRNA-016901 silencing attenuates irradiation-induced injury in bone mesenchymal stem cells via regulating the miR-1249-5p/HIPK2 axis.

Currently, bone marrow transplantation remains the basic treatment for various hematological tumors and irradiation is one of the most important pretreatment methods. However, irradiation pretreatment may result in damage to bone mesenchymal stem cells (BMSCs). The present study aimed to investigate the effect of circular RNA-016901 (circ-016901) on the injury of irradiation-induced BMSCs and the underlying mechanism. The expression levels of circ-016901, microRNA-1249-5p (miR-1249-5p) and homeodomain interacting protein kinase 2 (HIPK2) in irradiation-induced mouse BMSCs at various irradiation doses were detected via reverse transcription-quantitative PCR (RT-qPCR). The effect of circ-016901 on cell proliferation was examined using Cell Counting Kit-8 assays following silencing or overexpression of circ-016901. Cell apoptosis was detected by flow cytometry and caspase-3/7 activity. The expression of autophagy-related markers, including Beclin-1 and LC3-II/I, was detected at the mRNA and protein levels by RT-qPCR and western blotting, respectively. Irradiation treatment upregulated the expression of circ-016901 and HIPK2 and downregulated miR-1249-5p expression. The expression levels of LC3-II/I and Beclin-1 in BMSCs were downregulated in a dose-dependent manner. Silencing of circ-016901 promoted proliferation of irradiation-induced BMSCs and attenuated irradiation-induced apoptosis. Moreover, silencing of circ-016901 elevated the expressions of LC3-II/I and Beclin-1 in irradiation-induced BMSCs. Similar results were obtained with miR-1249-5p overexpression and HIPK2 silencing. These results demonstrated that circ-016901 silencing attenuated injury in irradiation-induced mouse BMSCs by regulating the miR-1249-5p/HIPK2 axis, providing a novel target for future research on the mechanism of radiation resistance in BMSCs.

Cell fate determined by the activation balance between PKR and SPHK1.

Double-stranded RNA (dsRNA)-dependent protein kinase R (PKR) activation via autophosphorylation is the central cellular response to stress that promotes cell death or apoptosis. However, the key factors and mechanisms behind the simultaneous activation of pro-survival signaling pathways remain unknown. We have discovered a novel regulatory mechanism for the maintenance of cellular homeostasis that relies on the phosphorylation interplay between sphingosine kinase 1 (SPHK1) and PKR during exogenous stress. We identified SPHK1 as a previously unrecognized PKR substrate. Phosphorylated SPHK1, a central kinase, mediates the activation of PKR-induced pro-survival pathways by the S1P/S1PR1/MAPKs/IKKα signal axis, and antagonizes PKR-mediated endoplasmic reticulum (ER) stress signal transduction under stress conditions. Otherwise, phosphorylated SPHK1 also acts as the negative feedback factor, preferentially binding to the latent form of PKR at the C-terminal kinase motif, inhibiting the homodimerization of PKR, suppressing PKR autophosphorylation, and reducing the signaling strength for cell death and apoptosis. Our results suggest that the balance of the activation levels between PKR and SPHK1, a probable hallmark of homeostasis maintenance, determines cell fate during cellular stress response.

[Effect of Mmu-circRNA_016901 on the Radiosensitivity of Bone Marrow Mesenchymal Stem Cells].

OBJECTIVE: To explore the effect of mmu-circRNA_016901 on the regulation of radiation injury of bone marrow stem cells and its mechanism. METHODS: Bone marrow stem cells were exposed to different dose of X-ray, then the expression level of mmu-circRNA_016901 in bone marrow cells treated with different doses of X-ray was detected. The luciferase reporter gene assay was used to confirm that miRNA1249-5p is the target of mmu-circRNA_016901, and RNA Binding Protein Immunoprecipitation was used to confirm that TGF-ß3 is the targeted on miRNA1249-5p，the expression of TGF-ß3 and cell proliferation were detected after the expression of mmu-circRNA_01690 was regulated. RESULTS: When the irradiation dose＜6 Gy, there were significant difference in the expression of mmn-circRNA-016901 after the bone marrow mesenchymal stem cells were treated by different doses of irradiation, which showed a statistically significant (P＜0.05). The luciferase reporter gene detection and co-immunoprecipitation experiments confirmed that Mmu-circRNA_016901 could binds to miRNA1249-5p specifically, and overexpression of mmu-circRNA_016901 could regulate miRNA1249-5p negatively, which resulted in a significant increase in TGF-ß3 expression and promoting of cell proliferation. CONCLUSION: mmu-circRNA_016901 affects the expression of TGF-ß3 through miRNA1249-5p, and thus participates in the regulation of the radiation damage mechanism of bone marrow mesenchymal stem cells.

Deoxynivalenol globally affects the selection of 3' splice sites in human cells by suppressing the splicing factors, U2AF1 and SF1.

Deoxynivalenol (DON) is one of the most abundant mycotoxins and has adverse effects on several biological processes, posing risks of protein synthesis-disrupting effects and ribotoxic response. Therefore, chronic exposure to DON would fundamentally reshape the global expression pattern. Whether DON causes toxic effects on mRNA splicing, a fundamental biological process, remains unclear. In this study, we found that administration of the relative low dosage of DON dramatically changed the alternative splicing of pre-mRNA in HepG2 cells. The overall number of transcripts with aberrant selection of 3' splice sites was significantly increased in DON-exposed HepG2 cells. This effect was further confirmed in two other human cell lines, HEK293 and Caco-2, suggesting that this DON-induced alteration in splicing patterns was universal in human cells. Among these DON-induced changes in alternative splicing, the expression levels of two related splicing factors, SF1 and U2AF1, which are essential for 3' splice site recognitions, were strongly suppressed. Overexpression of either of the two splicing factors strongly alleviated the DON-induced aberrant selection of 3' splice sites. Moreover, SF1 was required for human cell proliferation in DON exposure, and the restoration of SF1 expression partially reinstated the proliferation potential for DON-treated cells. In conclusion, our study suggests that DON, even at a low dosage, has great potential to change gene expression globally by affecting not only protein synthesis but also mRNA processing in human cells.

[Relation of Thromboelastography with Coagulation Function and Plt Count in Different Coagulation States].

OBJECTIVE: To investigate the correlation and consistency between thromboelastography (TEG) and routine tests, platelets count in different coagulation states (hypercoagulable, low coagulation, and normal coagulation) and to evaluate the clinical value of TEG, routine bloot test and Plt count. METHODS: The clinical data of 409 patients performed the TEG, coaglation 4 test and blood routine test in third Xiangya Hosptial of Central Sonth University from January 2015 to December 2017 were analyzed retrospectively. The TEG main parameters such as the activation time of clothing factor (R), the formation rate of blood clots (K), the maximal α-angle (Angle) and maximal amplitude (MA) were compared with prothrombin time (PT), activated partial thromboplastin time (APTT), fibrin (Fib), thrombin time (TT) in blood routine test as well as platelet (Plt) counts by using the person correlation, Kappa consistency and paired chi-square test in different coagulation states, at the same time the guiding rote of these 2 detection mathods for clinical application of blood was compared. RESULTS: R value positively correlated with PT, the correlation coefficient (r) under low, high and normal coagulation status was 0.376, 0.316 and 0.276 respectively (P<0.05); the Kruskal-Wallis test of pairwise comparison showed no statistical difference (P>0.05); the consistency of R with PT under low, high and normal coagalation status was 0.208, 0.227 and 0.131, respectively. The R value positively correlated with APTT, r value under low, high and normal coagulation status was 0.418, 0.258 and 0.458 respectively (P<0.05); the Kruskal Walls test of pairwise comparion showed that there was no difference between value of low and normal coagulation status (P>0.05), while there was difference between r value of low and high coagulation status (P<0.05), the consistences of R value with APTT under low, high and normal coagulatiom status was 0.338, 0.291 and 0.161, with statistical difference (P<0.05). K value negatively correlated with Fib, r value under low, high and mornal coagulation status was -0.611, -0.411 and 0.311 respectively (P<0.05); the Kruskal-Wallis test of pairwise comparison all showed that statistical difference (P<0.05); the consistencey of K value with Fib was 0.432, 0.481 and 0.323 respectively under low, high and normal coagulation states. K value negatively correlated with plt count, r value under low, high and normal coagulation status was -0.278, -0.238 and -0.278 respectively (P<0.05); the consistency of K value with Fib level under low,high and normal coagulation status was 0.401, 0.312 and 0.279 respectively(P<0.05). Angle postively correlated with Fib level, r value under low, high and normal coagulation status was 0.638, 0.538 and 0.438 respectively (P<0.05); the Kruskal-Wallis test of pairwise comparison showed the statistical difference (P<0.05); the consistency of Angle with under low,high and normal coagulation status Fib was 0.323, 0.357 and 0.288 respectively(P<0.05). MA value positively correlated with Fib level (r=0.351, 0.381 and 0.211, P<0.05); the Kruskal-Wallis test of pairwise comparison showed that there was no difference of r value under low- and high-coagulation states (P>0.05), while there were differences of r values under low- and high-coagulation states with normal coagulation (P<0.05); the consistency of MA with Fib level under 3 kinds of coagulation states was 0.510, 0.467 and 0.427 respectively (P<0.05). MA value positively correlated with Plt count (r=0.478, 0.515 and 0.378) respectively under 3 kinds of coagulation states; the Kruskal-Wallis test of pairuse comparison both showed the statistical difference (P<0.05); the consistency of MA with Plt count under 3 kinds of coagulation status was 0.581, 0.461 and 0.350 (P<0.05). CONCLUSION: The TEG correlates with results of blood coagulation test and Plt detection; the correlation and consistecy of TEG with routine blood coagalation test and Plt detection are different under different status. Therefore, for patients who possibly had pathologic hyper- and hypo-coagulation, the TEG detection should be performed, so as to dynamically monitor the blood coagulation in vivo, guide the rational use of drugs and blood transfusion, reduce the risk of embolion and blood transfusion.

Dandelion root extract protects NCM460 colonic cells and relieves experimental mouse colitis.

The protective potential of dandelion on acute hepatitis, lung injury and colorectal cancer has recently been revealed. Importantly, ulcerative colitis (UC), a clinically defined inflammatory bowel disease, accelerates the risk of colorectal cancer. However, studies focusing on the activity of dandelion on UC are extremely limited. In the present study, we found that an aqueous extract of dandelion root increases cell viability and decreases apoptosis in dextran sodium sulfate (DSS)-incubated NCM460 human colonic epithelial cells, probably through removing the production of reaction oxygen species and blocking nuclear factor-kappaB signaling. We then examined the anti-colitis efficacy of this extract in an in vivo study. We detected that dandelion root extract efficiently ameliorates progressive acute injury as demonstrated by a reduction in body weight loss, severity scores of disease index and shortened colon length during DSS treatment, as well as reducing the inflammatory conditions and oxidative stress in the colon of DSS-induced mice. Our study clearly demonstrates that dandelion has a strong cytoprotective effect on NCM460 colonocytes and shows powerful defense on an established experimental mouse model of DSS-induced UC. Therefore, dandelion root extract can be an effective anti-colitis complex mixture and can provide a complementary alternative to currently available therapeutic intervention in UC.

Adsorption and visible-light photodegradation of organic dyes with TiO2/conjugated microporous polymer composites.

A series of composite materials made of TiO2 and conjugated microporous polymers (CMPs) were prepared with a hydrothermal method and used as both adsorbents and photocatalysts for the adsorption and visible-light photodegradation of organic dyes in aqueous solutions. It is found that the blending of CMPs can significantly improve both the adsorption capacity and the photocatalytic degradation activity of TiO2 towards organic dyes.

MicroRNA-199a-5p inhibits VEGF-induced tumorigenesis through targeting oxidored-nitro domain-containing protein 1 in human HepG2 cells.

VEGF induces deterioration of hepatocellular carcinoma (HCC) by enhancing cell proliferation and migration. MicroRNAs regulate many cellular processes. In this study, we examined the regulation of tumorigenesis in HCC cells by microRNAs in relation to the effect of VEGF. Differences in microRNA expression between HepG2 and THLE-3 cells were characterized by microarray analysis. The results showed that miR-199a-5p expression was markedly downregulated in HepG2 cells and was inhibited in VEGF-overexpressing HepG2 cells in a dose- and time-dependent manner. This miRNA also inhibited cell proliferation and migration, as demonstrated by MTT and cell migration assays. Oxidored-nitro domain-containing protein 1 (NOR1), a nitroreductase, was identified as a downstream target gene of miR-199a-5p, and upregulation of NOR1 proved critical for the inhibition of VEGF-induced cell proliferation and migration in HepG2 cells by miR-199a-5p. These results indicate that miR-199a-5p is critical for regulating cell proliferation and migration by targeting and upregulating NOR1 in human HepG2 cells.

[Ratio of erythrocyte and plasma in massive blood transfusion].

This study was purposed to explore the suitable ratio between fresh frozen plasma and erythrocyte by retrospective analysis of coagulation in patients with massive blood transfusion. The clinical data of 151 cases with massive blood transfusion from January 2011 to January 2013 were analyzed retrospectively. According to coagulation, patients were divided into coagulation normal group (138 cases) and coagulation dysfunction group (13 cases). Based on the ratio of 1:1 of fresh frozen plasma and erythrocyte, the patients were divided into high plasma group(2:1), medium plasma group (1:1) and low plasma (<1:1) subgroups. Coagulation was detected before and after 24 h of massive blood transfusion. The results showed that prothrombin time (PT), activated partial thromboplastin time (APTT) and thrombin time (TT) were prolonged, fibrinogen (FIB) level decreased significantly (all P < 0.05) in the low plasma subgroup of coagulation normal group after massive blood transfusion 24 h; the high plasma and the medium plasma group of coagulation normal group had no significant changes in coagulation (P > 0.05); prothrombin time, activated partial thromboplastin time, thrombin time and fibrinogen level in the medium plasma and low plasma subgroup of coagulation dysfunction group after massive transfusion was still in abnormal levels (P > 0.05), coagulation function in high plasma subgroup was improved significantly (P < 0.05). It is concluded that the ratio of plasma to erythrocyte should be adjusted according to the patient's coagulation function during massive blood transfusion, the ratio between fresh frozen plasma and erythrocyte is recommended to be 2:1 in patients of coagulation dysfunction in order to improve the patient's coagulation function and to reduce the incidence of adverse event, the ratio of fresh frozen plasma to erythrocyte is recommended to be 1:1 in patients with normal coagulation so as to reduce the dilutional coagulopathy and hypervolemia of blood.

[Analysis on the effectiveness in 727 times of platelet transfusion].

This study was purposed to analyze the efficiency of platelet transfusion and to explore factors influencing platelet transfusion efficiency. 727 times of platelets transfusion in 254 patients in The Third Xiangya Hospital from September 2010 to May 2011 were analyzed retrospectively. Moreover, according to symptoms, times of platelet transfusion, blood types and splenomegaly, the corrected count of increment (CCI) and percentage of platelet recovery (PPR) were calculated for evaluation of platelet transfusion efficiency. The results showed that there were 456 effective transfusions out of 727 transfusions (62.72). Among them, the therapeutic effect of platelet transfusion for patients with acute blood loss anemia and chronic systemic diseases was relatively obvious, specially for chronic renal disease, the effective efficiency of them was 94.12. The patients with splenomegaly showed a significant impact on platelet transfusion efficiency (41.07). Analysis found that the frequency of platelet transfusion negatively correlated with transfusion efficiency. It is concluded that the transfusion frequency and splenomegaly are factors influencing the transfusion efficiency.