An integrative bioinformatics investigation and experimental validation of chromobox family in diffuse large B-cell lymphoma.

BACKGROUND: Diffuse large B-cell lymphoma (DLBCL) is one of the most aggressive malignant tumors. Chromobox (CBX) family plays the role of oncogenes in various malignancies. METHODS: The transcriptional and protein levels of CBX family were confirmed by GEPIA, Oncomine, CCLE, and HPA database. Screening of co-expressed genes and gene function enrichment analysis were performed by GeneMANIA and DAVID 6.8. The prognostic value, immune cell infiltration and drug sensitivity analysis of CBX family in DLBCL were performed by Genomicscape, TIMER2.0, and GSCALite database. Confirmatory Tests of CBX family protein expression in DLBCL were performed by immunohistochemistry. RESULTS: The mRNA and protein expressions of CBX1/2/3/5/6 were higher in DLBCL tissues than control groups. Enrichment analysis showed that the functions of CBX family were mainly related to chromatin remodeling, methylation-dependent protein binding, and VEGF signaling pathway. The high mRNA expressions of CBX2/3/5/6 were identified to be associated with short overall survival (OS) in DLBCL patients. Multivariate COX regression indicated that CBX3 was independent prognostic marker. Immune infiltration analysis revealed that the mRNA expressions of CBX family (especially CBX1, CBX5, and CBX6) in DLBCL were significantly correlated with the infiltration of most immune cells (including B cells, CD8 + T cells, CD4 + T cells, neutrophils, monocytes, macrophages, and Treg cells). Meanwhile, there was a strong correlation between the expression levels of CBX1/5/6 and surface markers of immune cells, such as the widely studied PVR-like protein receptor/ligand and PDL-1 immune checkpoint. Notably, our study found that DLBCL cells with CBX1 over-expression were resistant to the common anti-tumor drugs, but CBX2/5 had two polarities. Finally, we confirmed the higher expressions of CBX1/2/3/5/6 in DLBCL tissues compared with control groups by immunohistochemistry. CONCLUSION: We provided a detailed analysis of the relationship between the CBX family and the prognosis of DLBCL. Distinguished from other studies, We found that high mRNA expressions of CBX2/3/5/6 were associated with poor prognosis in DLBCL patients, and Multivariate COX regression indicated that CBX3 was independent prognostic marker. Besides, our study also found an association between the CBX family and anti-tumour drug resistance, and provided a relationship between CBX family expression and immune cell infiltration.

The benefits of autotrophic nitrogen removal from high concentration of urea wastewater through a process of urea hydrolysis and partial nitritation in sequencing batch reactor.

For the sake of high efficiency and saving operational cost for high-concentration urea wastewater treatment, a novel two-stage partial nitritation (PN)-anammox process containing simultaneous urea hydrolysis and PN in sequencing batch reactor (SBR) was investigated. Although the influent urea concentration increased from 500 to 1200 mg/L, the SBR simultaneously achieved urea removal efficiency higher than 98% and stable PN with effluent NO2--N/NH4+-N ratio of 1.0-1.3 without any extra alkalinity addition. The intracellular hydrolysis was the dominant mechanism for urea removal and persistent free ammonia inhibition on nitrite-oxidizing bacteria was the main reason for nitrite accumulation of 97.92% in SBR. The subsequent anammox reactor showed efficient nitrogen removal performance with average ammonium removal efficiency, nitrogen removal efficiency and maximum nitrogen removal loading rate of 98.08%, 81.45% and 1.05 kg N·m-3·d-1 respectively. High-throughput sequencing results indicated Gemmatimonadetes became the most abundant bacterial phylum related to potential intracellular urea hydrolysis and displayed obvious ammonium-oxidizing bacteria enrichment and nitrite-oxidizing bacteria inhibition in SBR, and the dominant anammox bacteria (Candidatus_Kuenenia) in anammox reactor. The proposed process was proven to be promising for high-concentration urea wastewater treatment, facilitating the sustainable development of the urea industry in the future.

Characterization of the transcriptional activation domains of human TEF3-1 (transcription enhancer factor 3 isoform 1).

TEF3-1 (transcription enhancer factor 3 isoform 1) is a human transcriptional factor, which has a N-terminal TEA/ATTS domain supposedly for DNA binding and C-terminal PRD and STY domains for transcriptional activation. Taking advantage of the efficient reporter design of yeast two-hybrid system, we characterized the TEF3-1 domains in activating gene expression. Previously study usually mentioned that the C-terminal domain of TEF3-1 has the transcriptional activity, however, our data shows that the peptides TEF3-11-66 and TEF3-1197-434 functioned as two independent activation domains, suggesting that N-terminal domain of TEF3-1 also has transcriptional activation capacity. Additionally, more deletions of amino acids 197-434 showed that only the peptides TEF3-1197-265 contained the minimum sequences for the C-terminal transcriptional activation domain. The protein structure is predicted to contain a helix-turn-helix structure in TEF3-11-66 and four ß sheets in TEF3-1197-265. Finally, after the truncated fragments of TEF3-1 were expressed in HUVEC cells, the whole TEF3-1 and the two activation domains could increase F-actin stress fiber, cell proliferation, migration and targeted gene expression. Further analysis and characterization of the activation domains in TEF3-1 may broaden our understanding of the gene involved in angiogenesis and other pathological processes.

Effective recovery of the nitritation process through hydrogen peroxide.

This study successfully achieved stable nitritation by adding hydrogen peroxide (H2O2) to the nitrification sludge whose nitritation stability had been destroyed. The batch experiment demonstrated that, the activity of ammonia-oxidizing bacteria (AOB) was restored more rapidly than that of nitrite oxidizing bacteria (NOB) after the addition of H2O2, thereby selectively promoting AOB enrichment and NOB washout. When the H2O2 concentration was 6.25 mg/L, the NOB activity was significantly reduced and the nitrite accumulation rate (NAR) was more than 95% after 18 cycles of nitrifying sludge restoration. As a result, H2O2 treatment enabled a nitrifying reactor to recover stable nitritation performance via H2O2 treatment, with the NAR and ammonia removal efficiency (ARE) both exceeding 90%. High-throughput sequencing analysis revealed that H2O2 treatment was successful in restoring nitritation, as the relative abundance of Nitrosomonas in the nitrifying reactor increased from 6.43% to 41.97%, and that of Nitrolancea decreased from 17.34% to 2.37%. Recovering nitritation by H2O2 inhibition is a low operational cost, high efficiency, and non-secondary pollution nitritation performance stabilization method. By leveraging the varying inhibition degrees of H2O2 on AOB and NOB, stable nitrification can be efficiently restored at a low cost and without causing secondary pollution.

[Nitritation Performance of Zeolite Moving Bed Biofilm Reactor for Ammonium Wastewater Treatment].

A new type of zeolite-suspended packing was developed by using zeolite as an important raw material, which was then used to start the zeolite moving bed biofilm reactor (ZMBBR). ZMBBR was compared with the ceramsite moving bed biofilm reactor (CMBBR) packed with ordinary ceramsite-suspended packing to investigate the different nitritation performance. The results showed that stable nitritation was successfully achieved in two reactors by the inhibitory effect of free ammonia (FA), and both of their nitrite accumulation rates (NAR) reached 90%; due to the adsorption of zeolite to ammonium, ZMBBR relieved the inhibition of FA on AOB faster than CMBBR and achieved nitritation earlier; CMBBR and ZMBBR could maintain long-term stable nitrosation when ρ(NH4+-N) was 350 mg·L-1 and 1050 mg·L-1 and NPRAVG was 0.43 kg·(m3·d)-1 and 1.26 kg·(m3·d)-1, respectively, and ARECMBBR=82.21% and AREZMBBR=88.85%. In the process of the influent ρ(NH4+-N) gradually increasing from 250 mg·L-1 to 1250 mg·L-1, the maximum nitrite production rate (NPR) of CMBBR was 0.5634 kg·(m3·d)-1; when ρ(FA) reached 166 mg·L-1 at the influent ρ(NH4+-N) of 750 mg·L-1, CMBBR broke down for the heavy inhibition of FA. The maximum NPR of ZMBBR was 1.800 kg·(m3·d)-1, and the performance of ZMBBR was getting worse after the ρ(FNA) reached the peak value of 1.9611 mg·L-1 at the influent ρ(NH4+-N) of 1250 mg·L-1. Subsequently, the ρ(FA) of ZMBBR reached 158 mg·L-1 rapidly, the NPR dropped significantly to 0.9028 kg·(m3·d)-1, and the performance of ZMBBR became significantly worse. It was demonstrated by high-throughput sequencing analysis that the dominant strain of ZMBBR and CMBBR was Nitrosomonas_europaea, and the relative abundances of N._europaea in ZMBBR and CMBBR were 11.15% and 10.92%, respectively.

Partial nitritation performance and microbial community in sequencing batch biofilm reactor filled with zeolite under organics oppression and its recovery strategy.

Influences of organics on partial nitritation performance were investigated in a lab-scale sequencing batch biofilm reactor filled with zeolite. Significant differences in nitrite production rate (NPR) were observed between different dosages of glucose. With influent COD/N ratio from 0 to 1.5, NPR declined from 0.4 to 0.05 kg/(m3·d). Meanwhile, an appropriate NO2--N/NH4+-N ratio (1.4 ± 0.5) could be obtained for simultaneous anammox denitrification at COD/N ratio of 0.5. Increasing airflow rate was found as an effective recovery strategy. Other than competition of heterotrophs with nitrifiers for dissolved oxygen, it has been verified that addition of organics generated higher free ammonia, and then further inhibitedammonium oxidizing bacteria (AOB). Moreover, three-dimensional excitation-emission matrix (3D-EEM) results revealed that protein-like and humic acid-like substances were the main components in extracellularpolymericsubstances (EPS). And high-throughput sequencing analysis demonstrated that the relative abundance of AOB decreased.

Performance and mechanism of urea hydrolysis in partial nitritation system based on SBR.

Urea hydrolysis in partial nitritation process forming nitrite and ammonia is advantageous to subsequent treatment with ANAMMOX for total nitrogen removal. In this study, stable partial nitritation for urea wastewater with urea increasing from 250 to 2000 mg L-1 were achieved in an aerobic SBR. Urea removal efficiency and nitrite accumulation percentage both kept above 98%, with nitrite production rate about 0.985 kg N·m-3·d-1. Urea hydrolysis mechanism in this aerobic system was described as, (1) massive urea in the bulk was absorbed into cell, (2) urea was hydrolyzed by intracellular urease inside cell, (3) produced ammonia then slowly diffused into the bulk through membrane, which is later converted by ammonia-oxidizing bacteria (AOB) into nitrite. Due to this mechanism, the activity of AOB could not be inhibited by high FA (free ammonia) value under high urea concentration condition while nitrite-oxidizing bacteria (NOB) remained to be inhibited. An uncultured genus belonging to poorly characterized phylum Gemmatimonadetes was found enriched in this process and became dominant genus. This genus was speculated to have same energy pathway like ureaplasma, by absorbing excessive urea from environment and utilize urea hydrolysis to generate energy. So it was believed to be responsible for urea hydrolysis mechanism mentioned above. This SBR showed stable partial nitritation and high urea removal efficiency for treating urea wastewater, which was obviously feasible as the pretreatment process for subsequent ANAMMOX.

Biological nitrogen removal via combined processes of denitrification, highly efficient partial nitritation and Anammox from mature landfill leachate.

The combined processes of pre-denitrification, highly efficient partial nitritation and Anammox were developed to treat mature landfill leachate. In the partial nitritation stage, an outstanding nitrite production rate (NPR) of approximately 1.506 kg·(m3 day)-1 of mature landfill leachate was achieved in a zeolite biological aerated filter (ZBAF) due to the inhibition of nitrite-oxidizing bacteria (NOB) by free ammonia (FA) and free nitrous acid (FNA). With respect to the nitrogen removal performance of the combined process, remarkable nitrogen removal efficiencies (NRE) and nitrogen removal rates (NRR), which exceeded 90.0% and 0.490 kg·(m3 day)-1, respectively, were detected based on the stable and efficient partial nitritation performance and reasonable control of effluent nitrite to ammonium ratios (at approximately 1.2) in the ZBAF. High-throughput sequencing analysis further revealed that the dominant bacteria genera Paracoccus and Comamonas in the denitrification reactor, Nitrosomonas in the ZBAF and Candidatus Kuenenia and Candidatus Anammoxoglobus in the Anammox reactor were demonstrated to be responsible for denitrification, partial nitritation and Anammox process, respectively.

Nuclear localization of TEF3-1 promotes cell cycle progression and angiogenesis in cancer.

TEF3-1 (transcriptional enhancer factor 3 isoform 1), also known as TEAD4 (TEA domain family member 4), was recently revealed as an oncogenic character in cancer development. However, the underlying molecular pathogenic mechanisms remain undefined. In this paper, we investigated nuclear TEF3-1 could promote G1/S transition in HUVECs, and the expression levels of cyclins and CDKs were upregulated. Additionally, if TEF3-1 was knocked down, the expression of cyclins and CDKs was downregulated while the expression of P21, a negative regulator of the cell cycle, was upregulated. A microarray analysis also confirmed that TEF3-1 overexpression upregulates genes that are related to cell cycle progression and the promotion of angiogenesis. Moreover, we observed that nuclear TEF3-1 was highly expressed during the formation of vascular structures in gastric cancer (GC). Finally, tumor xenograft experiments indicated that, when TEF3-1 was knocked down, tumor growth and angiogenesis were also suppressed. Taken together, these results demonstrate for the first time that TEF3-1 localization to the nucleus stimulates the cell cycle progression in HUVECs and specifically contributes to tumor angiogenesis. Nuclear TEF3-1 in HUVECs may serve as an oncogenic biomarker, and the suppression of TEF3-1 may be a potential target in anti-tumor therapy.