Vitamin D receptor induces oxidative stress to promote esophageal squamous cell carcinoma proliferation via the p53 signaling pathway.

Background: Esophageal squamous cell carcinoma (ESCC) is a common pathological esophageal cancer with poor prognosis. Vitamin D deficiency reportedly occurs in ESCC patients, and this is related to single nucleotide polymorphism of vitamin D receptor (VDR). Objective: We investigated the effect of VDR on ESCC proliferation, invasion, and metastasis and its potential mechanism. Methods: ESCC and normal tissues were collected from 20 ESCC patients. The ESCC tissue microarray contained 116 pairs of ESCC and normal tissues and 73 single ESCC tissues. VDR expression and its clinicopathological role were determined by real-time quantitative polymerase chain reaction, Western blot, and immunohistochemistry staining. sh-VDR and VDR overexpression were used to validate the effect of VDR on ESCC cell phenotype, and tandem mass tag-based quantitative proteomics and bioinformatics methods identified differential VDR-related proteins. The downstream pathway and regulatory effect were analyzed using ingenuity pathway analysis (IPA). Differentially expressed proteins were verified through parallel reaction monitoring and Western blot. In vivo imaging visualized subcutaneous tumor growth following tail vein injection of VDR-deficient ESCC cells. Results: High VDR expression was observed in ESCC tissues and cells. Gender, T stage, and TNM stage were related to VDR expression, which was the independent prognostic factor related to ESCC. VDR downregulation repressed ESCC cell proliferation, invasion, and migration in vitro and subcutaneous tumor growth and lung metastases in vivo. The cell phenotype changes were reversed upon VDR upregulation, and differential proteins were mainly enriched in the p53 signaling pathway. TP53 cooperated with ABCG2, APOE, FTH1, GCLM, GPX1, HMOX1, JUN, PRDX5, and SOD2 and may activate apoptosis and inhibit oxidative stress, cell metastasis, and proliferation. TP53 was upregulated after VDR knockdown, and TP53 downregulation reversed VDR knockdown-induced cell phenotype changes. Conclusions: VDR may inhibit p53 signaling pathway activation and induce ESCC proliferation, invasion, and metastasis by activating oxidative stress.

Secure data storage based on blockchain and coding in edge computing.

Edge computing is an important tool for smart computing, which brings convenience to data processing as well as security problems. In particular, the security of data storage under edge computing has become an obstacle to its widespread use. To solve the problem, the mechanism combing blockchain with regeneration coding is proposed to improve the security and reliability of stored data under edge computing. Our contribution is as follows. 1) According to the three-tier edge computing architecture and data security storage requirements, we proposed hybrid storage architecture and model specifically adapted to edge computing. 2) Making full use of the data storage advantages of edge network devices and cloud storage servers, we build a global blockchain in the cloud service layer and local blockchain is built on the terminals of the Internet of things. Moreover, the regeneration coding is utilized to further improve the reliability of data storage in blockchains. 3) Our scheme provides a mechanism for periodically validating hash values of data to ensure the integrity of data stored in global blockchain.

Artemisia capillaris inhibited enterovirus 71-induced cell injury by preventing viral internalization.

Artemisia capillaris (A. capillaris) is a common herbal drug used for thousands years in ancient China. A. capillaris has been empirically used to manage hand-foot-mouth disease (HFMD), which is commonly caused by enterovirus 71 (EV71). EV71 can cause meningoencephalitis with mortality and neurologic sequelae without effective management. It is presently unknown whether A. capillaris is effective against EV71 infection. To test the hypothesis that it could protect cells from EV71-induced injury, a hot water extract of A. capillaris was tested in human foreskin fibroblast cells (CCFS-1/KMC) and human rhabdomyosarcoma cells (RD cells) by plaque reduction assay and flow cytometry. Inhibition of viral replication was examined by reverse quantitative RT-PCR (qRT-PCR). Its effect on translations of viral proteins (VP0, VP1, VP2, protease 2B and 3AB), and apoptotic proteins were examined by western blot. A. capillaris was dose-dependently effective against EV71 infection in both CCFS-1/KMC cells and RD cells by inhibiting viral internalization. However, A. capillaris was minimally effective on viral attachment, VP2 translation, and inhibition of virus-induced apoptosis. Further isolation of effective molecules is needed. In conclusion, A. capillaris has anti-EV71 activity mainly by inhibiting viral internalization. A. capillaris would be better to manage EV71 infection in combination with other agents.

[Comparisons of Microbial Numbers, Biomasses and Soil Enzyme Activities Between Paddy Field and Drvland Origins in Karst Cave Wetland].

The purpose of this study is to compare microbial number, microbial biomass as well as soil enzyme activity between paddy field and dryland originated karst wetland ecosystems. The soil samples (0-20 cm) of uncultivated wetland, paddy field and dryland were collected in Huixian karst cave wetland, Guilin, China. Microbial numbers and biomass were detected using dilute plate incubation counting and chloroform fumigation-extraction, respectively. Microbial DNA was extracted according to the manufacturer's instructions of the kit. Microbial activity was examined using soil enzyme assays as well. The result showed that the bacteria number in paddy filed was (4.36 +/- 2.25) x 10(7) CFU x g(-1), which was significantly higher than those in wetland and dryland. Fungi numbers were (6.41 +/- 2.16) x 10(4) CFU x g(-1) in rice paddy and (6.52 +/- 1.55) x 10(4) CFU x g(-1) in wetland, which were higher than that in dryland. Actinomycetes number was (2.65 +/- 0.72) x 10(6) CFU x g(-1) in dryland, which was higher than that in wetland. Microbial DNA concentration in rice paddy was (11.92 +/- 3.69) microg x g(-1), which was higher than that in dryland. Invertase activity was (66.87 +/- 18.61) mg x (g x 24 h)(-1) in rice paddy and alkaline phosphatase activity was (2.07 +/- 0.99) mg x (g x 2 h)(-1) in wetland, both of which were higher than those in dryland. Statistical analysis showed there was a significant positive correlation of microbial DNA content, alkaline phosphatase activity and microbial carbon with soil pH, soil organic carbon (SOC), total nitrogen, alkali-hydrolyzable nitrogen, soil moisture, exchangeable Ca2+ and exchangeable Mg2+, as well as a significant positive correlation of intervase activity with the former three microbial factors. The above results indicated that microbial biomass and function responded much more sensitively to land-use change than microbial number in karst cave wetland system. Soil moisture, SOC and some factors induced by land-use change could affect mainly microbiological characteristics. We suggest that rice paddy, a kind of constructed wetland, should be kept and protected in experimental area or buffer area of Huixian karst cave wetland in the light of its similar property with the natural wetland.

Areca nut extract induces pyknotic necrosis in serum-starved oral cells via increasing reactive oxygen species and inhibiting GSK3ß: an implication for cytopathic effects in betel quid chewers.

Areca nut has been proven to be correlated with various pathologic alterations in oral cavity. However, the mechanisms for such cytopathic effects are still elusive due mostly to the limitations of cell culture systems. Here we discovered that areca nut extract (ANE) induced production of autophagosome vacuoles in cells cultured with rich medium but induced pyknosis and ballooning, two morphological alterations frequently observed in betel quid chewers, in cells under a serum-free culture condition. Permeability of the serum-starved cells to propidium iodide (PI) confirmed ANE induced novel necrosis with pyknosis (pyknotic necrosis), providing a possible explanation for inflammatory infiltration in chewers' mucosa. In these serum-starved cells, ANE strongly induced reactive oxygen species (ROS), which acted as a key switch for the initiation of pyknotic necrosis. Calcium flux was also involved in the morphological alterations. Besides, inhibition of GSK3ß by SB216763 significantly exacerbated the pyknotic necrosis either induced by ANE or H2O2 in serum-starved cells, suggesting that GSK3ß is a critical regulator for ANE/ROS-mediated pyknotic necrosis. Interestingly, LC3-II transition and PARP cleavage were still detected in the serum-starved cells after ANE treatment, suggesting concurrent activation of apoptotic and autophagic pathways. Finally, insulin could counteract the effect of ANE-induced pyknotic necrosis. Taken together, these data provide a platform for studying ANE-induced cytopathogenesis and the first clinical implication for several pathological alterations, such as ballooning and inflammatory infiltration, in betel quid chewers.

Arecoline downregulates levels of p21 and p27 through the reactive oxygen species/mTOR complex 1 pathway and may contribute to oral squamous cell carcinoma.

Arecoline, the major alkaloid of areca nut, has been shown to cause strong genotoxicity and is considered a potential carcinogen. However, the detailed mechanism for arecoline-induced carcinogenesis remains obscure. In this study, we noticed that the levels of p21 and p27 increased in two oral squamous cell carcinoma cell lines with high confluence. Furthermore, when treated with arecoline, elevated levels of p21 and p27 could be downregulated through the reactive oxygen species/mTOR complex 1 (ROS/mTORC1) pathway. Although arecoline decreased the activity of mTORC1, the amounts of autophagosome-like vacuoles or type II LC3 remained unchanged, suggesting that the downregulation of p21 and p27 was independent of autophagy-mediated protein destruction. Arecoline also caused DNA damage through ROS, indicating that the reduced levels of p21 and p27 might facilitate G (1) /S transition of the cell cycle and subsequently lead to error-prone DNA replication. In conclusion, these data have provided a possible mechanism for arecoline-induced carcinogenesis in subcytolytic doses in vivo.

Reversine suppresses oral squamous cell carcinoma via cell cycle arrest and concomitantly apoptosis and autophagy.

BACKGROUND: The effective therapies for oral cancer patients of stage III and IV are generally surgical excision and radiation combined with adjuvant chemotherapy using 5-Fu and Cisplatin. However, the five-year survival rate is still less than 30% in Taiwan. Therefore, evaluation of effective drugs for oral cancer treatment is an important issue. Many studies indicated that aurora kinases (A, B and C) were potential targets for cancer therapies. Reversine was proved to be a novel aurora kinases inhibitor with lower toxicity recently. In this study, the potentiality for reversine as an anticancer agent in oral squamous cell carcinoma (OSCC) was evaluated. METHODS: Effects of reversine on cell growth, cell cycle progress, apoptosis, and autophagy were evaluated mainly by cell counting, flow cytometry, immunoblot, and immunofluorescence. RESULTS: The results demonstrated that reversine significantly suppressed the proliferation of two OSCC cell lines (OC2 and OCSL) and markedly rendered cell cycle arrest at G2/M stage. Reversine also induced cell death via both caspase-dependent and -independent apoptosis. In addition, reversine could inhibit Akt/mTORC1 signaling pathway, accounting for its ability to induce autophagy. CONCLUSIONS: Taken together, reversine suppresses growth of OSCC via multiple mechanisms, which may be a unique advantage for developing novel therapeutic regimens for treatment of oral cancer in the future.