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Abstract Background Our recent cDNA microarray data showed that centromere protein F (CENP-F) is significantly upregulated in primary cultured nasopharyngeal carcinoma (NPC) tumor cells compared with normal nasopharyngeal epithelial cells. The goal of this study was to further investigate the levels of CENP-F expression in NPC cell lines and tissues to clarify the clinical significance of CENP-F expression in NPC as well as the potential therapeutic implications of CENP-F expression. Methods Real-time RT-PCR and western blotting were used to examine CENP-F expression levels in normal primary nasopharyngeal epithelial cells (NPEC), immortalized nasopharyngeal epithelial cells and NPC cell lines. Levels of CENP-F mRNA were determined by real-time RT-PCR in 23 freshly frozen nasopharyngeal biopsy tissues, and CENP-F protein levels were detected by immunohistochemistry in paraffin sections of 202 archival NPC tissues. Statistical analyses were applied to test for prognostic associations. The cytotoxicities of CENP-F potential target chemicals, zoledronic acid (ZOL) and FTI-277 alone, or in combination with cisplatin, in NPC cells were determined by the MTT assay. Results The levels of CENP-F mRNA and protein were higher in NPC cell lines than in normal and immortalized NPECs. CENP-F mRNA level was upregulated in nasopharyngeal carcinoma biopsy tissues compared with noncancerous tissues. By immunohistochemical analysis, CENP-F was highly expressed in 98 (48.5%) of 202 NPC tissues. Statistical analysis showed that high expression of CENP-F was positively correlated with T classification (P < 0.001), clinical stage (P < 0.001), skull-base invasion (P < 0.001) and distant metastasis (P = 0.012) inversely correlated with the overall survival time in NPC patients. Multivariate analysis showed that CENP-F expression was an independent prognostic indicator for the survival of the patient. Moreover, we found that ZOL or FTI-277 could significantly enhance the chemotherapeutic sensitivity of NPC cell lines (HONE1 and 6-10B) with high CENP-F expression to cisplatin, although ZOL or FTI-277 alone only exhibited a minor inhibitory effect to NPC cells. Conclusion Our data suggest that CENP-F protein is a valuable marker of NPC progression, and CENP-F expression is associated with poor overall survival of patients. In addition, our data indicate a potential benefit of combining ZOL or FTI-277 with cisplatin in NPC suggesting that CENP-F expression may have therapeutic implications.

Abstract Background Many kinetochore proteins have been shown to be associated with human cancers. The aim of the present study was to clarify the expression of Centromere protein H (CENP-H), one of the fundamental components of the human active kinetochore, in esophageal carcinoma and its correlation with clinicopathological features. Methods We examined the expression of CENP-H in immortalized esophageal epithelial cells as well as in esophageal carcinoma cells, and in 12 cases of esophageal carcinoma tissues and the paired normal esophageal tissues by RT-PCR and Western blot analysis. In addition, we analyzed CENP-H protein expression in 177 clinicopathologically characterized esophageal carcinoma cases by immunohistochemistry. Statistical analyses were applied to test for prognostic and diagnostic associations. Results The level of CENP-H mRNA and protein were higher in the immortalized cells, cancer cell lines and most cancer tissues than in normal control tissues. Immunohistochemistry showed that CENP-H was expressed in 127 of 171 ESCC cases (74.3%) and in 3 of 6 esophageal adenocarcinoma cases (50%). Statistical analysis of ESCC cases showed that there was a significant difference of CENP-H expression in patients categorized according to gender (P = 0.013), stage (P = 0.023) and T classification (P = 0.019). Patients with lower CENP-H expression had longer overall survival time than those with higher CENP-H expression. Multivariate analysis suggested that CENP-H expression was an independent prognostic marker for esophageal carcinoma patients. A prognostic value of CENP-H was also found in the subgroup of T3~T4 and N0 tumor classification. Conclusion Our results suggest that CENP-H protein is a valuable marker of esophageal carcinoma progression. CENP-H might be used as a valuable prognostic marker for esophageal carcinoma patients.

BACKGROUND Soil limit water content is an important basis for fine-grained soil classification and engineering property evaluation. Additionally, it is an important evaluation index for clay exploration and industrial utilization. Inaccurate test results of soil limit water content may lead to serious engineering safety accidents and personal and economic property losses. Certified reference materials (CRMs) are important for ensuring the accuracy, comparability, and effectiveness of the analyzed data. However, no certified reference materials for soil limit water content exist; therefore, the preparation of CRMs for the soil limit water content holds great significance. OBJECTIVES To prepare CRMs of soil limit water content. METHODS In strict accordance with the specifications and standards, such as "Technical Specifications for First-Class Reference Materials" (JJF 1006-1994) and "General Principles and Statistical Principles for the Valuation of Reference Materials" (JJF 1343-2012), five standard materials of soil limit water content (GBW07969, GBW07970, GBW07971, GBW07972, and GBW07973) have been developed. These samples were collected from Huaibei of Anhui province, Datong of Shanxi province, and Nanjing of Jiangsu province. After artificial crushing, drying, and sterilization, the samples were finely crushed to less than 0.25mm by a large ball mill. After particle size analysis, the samples were bottled and numbered in a clean room. RESULTS Twenty-five bottles of each sample were randomly selected for homogeneity testing. All the measured values of F were less than F0.05 (24, 25)=1.96, and the relative standard deviation (RSD) was between 1.16% and 2.67%, which indicated good uniformity. There were no significant differences in the long-term stability test (12months) and the short-term stability test (60℃, -20℃). The certified values of 10mm liquid limit, plastic limit, and plasticity index were 26.3%-39.9%, 16.9%-22.2%, and 10.0%-17.7%, respectively. The gradient series was significant, which included silty clayey and clayey type of soils. CONCLUSIONS Five first classes of National CRMs (GBW07969, GBW07970, GBW07971, GBW07972, and GBW07973) of soil limit water content were successfully prepared. This series of CRMs can be used for calibration of instruments and equipment, quality control, capability verification, and other technical quality activities, which provide a guarantee for the accuracy requirements of soil limit water test data in hydraulic environment geological exploration, geotechnical engineering exploration, clay mine exploration, and other related disciplines.

The Laser Particle Size Analyzer is being more widely used due to its fast test speed, convenient operation and good repeatability, to name a few of its qualities. However technical parameters of instrument and test conditions affect the results significantly. The test conditions of the HELOS/BF Laser Particle Size Analyzer were optimized by experiments described in this paper. The results indicate that samples with light specific-gravity or small particle size require lower stirring speed, higher ultrasonic intensity and longer ultrasonic time. Samples with heavy specific-gravity or large particle size require higher stirring speed, lower ultrasonic intensity and shorter ultrasonic time. The dispersant can accelerate the dispersion of the sample particles, especially for the small particle size samples. The low injection concentration (shading rate) with less particles leads to higher measured results for particle size because the weak signal causes bad representation for the samples. The high injection concentration leads to lower measured results based on the broad range of particle size distribution by multiple scattering. It suggests that the injection sample concentration (shading rate) between 10%-25% was appropriate for the HELOS/BF Laser Particle Size Analyzer.

Inflammatory cytokine interleukin-17 (IL-17) has been associated with the risk of progressive cancers including lung cancer. However, it remains unclear how IL-17 may contribute to the invasion and development of these inflammation-associated malignancies. Here we aimed to investigate the role of IL-17 in lung cancer cell development. Epithelial-mesenchymal transition (EMT) has been recently proposed as a developmental process which plays an important role in cancer progression and metastases. Here we show that IL-17 might promote EMT in lung cancer cells by inducing the transcriptional repressor ZEB1. Exposure to IL-17 upregulated the signature EMT phenotypic markers vimentin and E-cadherin in lung cancer cells, and compared with controls, increased cell migration was observed in IL-17-treated lung cancer cells. ZEB1 mRNA and protein expression was induced by IL-17, and IL-17 stimulated nuclear localization of phosphorylated ZEB1. Conversely, suppressing ZEB1 expression by ZEB1 siRNA abrogated IL-17-stimulated vimentin expression and cell migration. Moreover, the phosphorylation of IκBα was required for IL-17-induced expression of ZEB1, suggesting the involvement of canonical NF-κB signaling. To check this hypothesis, we used IKK inhibitor BAY 11-7028 to block NF-κB activity. We found that BAY 11-7028 abrogated IL-17-induced ZEB1 expression, cell migration, and EMT, thus confirming that NF-κB is required for IL-17 to induce these aggressive phenotypes in lung cancer cells. Taken together, our data support the idea that IL-17-induced EMT promotes lung cancer cell migration and invasion via NF-κB-mediated upregulation of ZEB1. This study reveals a new signaling axis through which the tumor microenvironment causes ZEB1 expression to promote cancer metastasis. We suggest that targeting IL-17-induced ZEB1 expression may offer an effective therapeutic strategy for lung cancer treatment.

Aim: To explore the synergistic effect of MBP 68-86 and 87-99, on the inhibition of experimental autoimmune encephalomyelitis (EAE) in Lewis rat by nasal administration., Methods: Three different MBP peptides(MBP 68-86, 87-99, and the non-encephalitogenic peptide 110-128) were synthesized and administrated nasally to Lewis rat on day-11, -10, -9, -8 and -7 prior to immunization with the guinea pig MBP (gp-MBP)+CFA, which was used to induce EAE. The protective effect on Lewis rat from EAE by the MBP peptides was evaluated., Results: Protection was achieved with the encephalitogenic peptides MBP 68-86 and 87-99, MBP 68-86 being more potent, but not with MBP 110-128. Neither MBP 68-86 nor 87-99 used alone conferred complete protection to gp-MBP-induced EAE. In contrast, nasal administration of a mixture of MBP 68-86 and 87-99 completely blocked gp-MBP-induced EAE even at lower dosage than being used alone. Rats tolerized with MBP 68-86+87-99 nasally showed decreased T cell responses to MBP, reflected by lymphocyte proliferation and IFN-gamma ELISPOT assays. Rats tolerized with MBP 68-86+87-99 also had abrogated MBP-reactive IFN-gamma and TNF-alpha mRNA expression in lymph node cells compared to rats receiving MBP 110-128 nasally, while similar low levels of MBP-reactive TGF-beta and IL-4 mRNA expressing cells were observed in the two groups., Conclusion: Nasal administration of encephalitogenic MBP peptides can induce antigen-specific T cell tolerance and confer incomplete protection to gp-MBP-induced EAE, and MBP 68-86 and 87-99 have synergistic effects. Non-regulatory mechanisms are proposed to be responsible for tolerance development after nasal peptide administration.

Aim: To explore the mechanism of differentiation of bone marrow stromal cells (BMSCs) into neurons in different micro-environments in vitro., Methods: BMSCs were isolated from bone marrow of SD rats and cultured and expanded in vitro. After being identified by immunofluorescence staining, the BMSCs labeled with PKH67 were co-cultured with foetal brain neural cells in the same plate well or in two-layer Petri dish. 8 days later, the BMSCs were detected by immunofluorescence staining., Results: After being co-cultured with foetal brain neural cells at the same time, some BMSCs differentiated into neurons. (32.72+/-2.56)% of the BMSCs expressed neuron-specific enolase (NSE) in the co-cultured group, which was obviously much more than that in control group (P <0.05). Only (4.87+/-0.79)% of the BMSCs expressed NSE when the BMSCs co-cultured with foetal brain neural cells in two-layer Petri dish, which had no difference with the control group (P>0.05). The number of differentiated BMSCs was less than that of the co-cultured group (P <0.05)., Conclusion: In vitro, the local microenvironment formed by neural cells can promote BMSCs to differentiate into neurons, and close contact between BMSCs and neural cells is an important condition that induce BMSC to differentiate into neurons.