Li Hong Yin

Isolation and Characterization of an Algicidal Bacterium Indigenous to Lake Taihu with a Red Pigment able to Lyse Microcystis aeruginosa

OBJECTIVEnTo isolate and characterize indigenous algicidal bacteria and their algae-lysing compounds active against Microcystis aeruginosa, strains TH1, TH2, and FACHB 905.nnnMETHODSnThe bacteria were identified using the Biolog automated microbial identification system and 16S rDNA sequence analysis. The algae-lysing compounds were isolated and purified by silica gel column chromatography and reverse-phase high performance liquid chromatography. Their structures were confirmed by Nuclear Magnetic Resonance (NMR) and Fourier Transform Infrared (FT-IR) spectroscopy. Algae-lysing activity was observed using microscopy.nnnRESULTSnThe algae-lysing bacterium LTH-2 isolated from Lake Taihu was identified as Serratia marcescens. Strain LTH-2 secreted a red pigment identified as prodigiosin (C20H25N3O), which showed strong lytic activity with algal strains M. aeruginosa TH1, TH2, and FACHB 905 in a concentration-dependent manner. The 50% inhibitory concentration (IC50) of prodigiosin with the algal strains was 4.8 (± 0.4)× 10⁻² μg/mL, 8.9 (± 1.1)× 10⁻² μg/mL, and 1.7 (± 0.1)× 10⁻¹ μg/mL in 24 h, respectively.nnnCONCLUSIONnThe bacterium LTH-2 and its pigment had strong Microcystis-lysing activity probably related to damage of cell membranes. The bacterium LTH-2 and its red pigment are potentially useful for regulating blooms of harmful M. aeruginosa.

Integrated analysis of competing endogenous RNA network revealing lncRNAs as potential prognostic biomarkers in human lung squamous cell carcinoma

Accumulating evidence shows the important role of long non-coding RNAs (lncRNAs) in competing endogenous RNA (ceRNA) networks for predicting survival in tumor patients. However, prognostic biomarkers for lung squamous cell carcinoma (LUSC) are still lacking. The objective of this study is to identify a lncRNA signature for evaluation of overall survival (OS) in 474 LUSC patients from The Cancer Genome Atlas (TCGA) database. A total of 474 RNA sequencing profiles in LUSC patients with clinical data were obtained, providing a large sample of RNA sequencing data, and 83 LUSC-specific lncRNAs, 26 miRNAs, and 85 mRNAs were identified to construct the ceRNA network (fold change>2, P<0.05). Among these above 83 LUSC-specific lncRNAs, 22 were assessed as closely related to OS in LUSC patients using a univariate Cox proportional regression model. Meanwhile, two (FMO6P and PRR26) of the above 22 OS-related lncRNAs were identified using a multivariate Cox regression model to construct a risk score as an independent indicator of the prognostic value of the lncRNA signature in LUSC patients. LUSC patients with low-risk scores were more positively correlated with OS (P<0.001). The present study provides a deeper understanding of the lncRNA-related ceRNA network in LUSC and suggests that the two-lncRNA signature could serve as an independent biomarker for prognosis of LUSC.

Combined Effects between Functionalized Multi-Walled Carbon Nanotubes and Cigarette Smoke on Human Bronchial Epithelial Cells

To evaluate the combined cytotoxicity effects between functionalized multi-walled carbon nanotubes (MWCNTs-PC) and cigarette smoke solution (CSS), 16-HBE cells was used as the target cells and exposed to various concentrations of MWCNTs-PC and CSS combined together. Cell proliferation, cell apoptosis, DNA damage were detected by Methyl thiazolyl tetrazolium (MTT) assay, flow cytometry, single cell gel electrophoresis assay (SCGE) and micronuclear assay, respectively. The dose-dependent cytotoxic and genetic effects of CSS were found in our study. However, compared to the control group, the MWCNTs-PC exposed groups showed no significant difference in all concentration, with or without CSS exposure. It suggests that the MWCNTs-PC did not influence cellular toxicity or DNA damage of CSS on 16-HBE cells. No combined cytotoxic effects between NWCNTs-PC and CSS were found in this study.

Neurotoxicity Evaluation of Chlorpyrifos Exposure with Caenorhabditis elegans

Chlorpyrifos, a broad spectrum orgnaophosphorus insecticide, is extensively used in agricultural and household insect control, but the potential health effects associated human exposure to low levels is unclear, which has been subject of increasing concern in the last years. However, few studies about chlorpyrifos toxicity have been conducted in the model organism Caenorhabditis elegans, which can complement both in vitro and in vivo mammalian models in toxicology. In the present study, Caenorhabditis elegans was chosen to evaluate the neurotoxicity of chlopyrifos with 0.003, 0.03, 0.3, and 3 mg/L after 4h exposure by analyzing basic movements, feeding ability and inhibition of acetylcholinesterase activity. Results indicated that forward turn frequency and acetylcholinesterase activity were reduced significantly compared with control at the highest concentration group, while feeding ability showed to decrease significantly at the lowest concentration group. The impact of feeding ability seems more sensitive than other targets. Moreover, we assessed the postexposure recovery of basic movements after the 4h exposure of Chlorpyrifos. Forward turn frequence of worms exposed to the highest concentration exposure failed to recovery, so that this suggested chlorpyrifos may have the long term health effect at high concentration exposure. Results also demonstrated that Caenorhabditis elegans could be a good model for chlorpyrifos-induced toxic effects research.

Comprehensive analysis of a novel lncRNA profile reveals potential prognostic biomarkers in clear cell renal cell carcinoma

Clear cell renal cell carcinoma (ccRCC) is the main subtype of malignant kidney cancer. Long non‑coding RNA (lncRNA) serves a key role in predicting survival in patients with cancer. The present study aimed to develop an lncRNA‑related signature of prognostic values for patients with ccRCC. RNA sequencing data of 454xa0patients were analyzed from The Cancer Genome Atlas (TCGA). To identify the differentially expressed lncRNAs, the patients from four groups classified by tumor stages were compared. The association between survival outcome and lncRNA expression profile was assessed by the univariate and multivariate Cox proportional hazards model. Survival was analyzed using the log‑rank test, and functions of target lncRNAs were investigated through Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway analysis. Finally, 19xa0lncRNAs were identified as significantly associated with overall survival (OS) time. These lncRNAs were gathered as a signal prognostic signature, which may be a potential biomarker for the prognosis of ccRCC. The risk score was built to evaluate the predictive value of the lncRNA signature. There was a significant positive correlation between ccRCC patients with the low‑risk score and OS time (P<0.001). Reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR) was used to verify the result in 17xa0pairs of ccRCC and adjacent non‑tumor tissues. Functional enrichment analysis revealed that these lncRNAs were associated with several molecular pathways of the tumor. The RT‑qPCR validation was consistent with the TCGA bioinformatics results. In conclusion, a tumor‑specific lncRNA signature of 19xa0lncRNAs was identified and the joint prognostic power was evaluated in the present study, and this signature was determined to be a potential biomarker for the prognosis of ccRCC.

Identification and functional characterization of long non-coding RNAs in human gastric cancer

Abnormal regulation of long non-coding RNAs (lncRNAs) appears to be a primary feature of numerous types of human cancer. However, the association between the dysregulation of lncRNAs and functional alterations in gastric cancer (GC) remains unclear. In previous studies, we applied microarray and bioinformatics analyses to screen for key lncRNAs from the tumor tissues and matched adjacent non-tumor tissues of 10 patients with GC. There were seven key lncRNAs demonstrated to be significantly different between carcinoma tissues and adjacent non-tumor tissues. In the present study, the expression of these seven selected lncRNAs were validated in 82 patients with GC to further investigate the association between lncRNAs and GC clinical characterization. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) results demonstrated that RP5-919F19, MCPH1 antisense RNA 1 (CTD-2541M15) and urothelial carcinoma-associated 1 (UCA1) exhibited consistent upregulation in cancer compared with adjacent non-tumor tissues, whereas AP000459, LOC101928316, tumor suppressor candidate 8 (LINC01071) and maternally expressed 3 (MEG3) showed consistent downregulation. The results from the microarray and RT-qPCR experiments achieved 100% agreement. A correlation analysis indicated that RP5-919F19, LOC101928316 and MEG3 were significantly associated with tumor differentiation degree, RP5-919F19, UCA1 and MEG3 were significantly associated with lymph node metastasis, and RP5-919F19, CTD-2541M15 and UCA1 were significantly associated with tumor-node-metastasis stage (P<0.05). In addition, it was identified that the differential expression of LINC01071 and LOC101928316 significantly correlated with the age and gender of the GC patients, respectively (P<0.05). The results suggest that the lncRNAs RP5-919F19, LOC101928316, CTD-2541M15, UCA1 and MEG3 are closely associated with the invasion and metastasis of GC, which reveals these indicators as potential specificity biomarkers for the diagnosis, prognosis and classification of GC. Thus, these lncRNAs merit further study as novel candidate biomarkers for the clinical diagnosis of GC and as potential targets for therapy.

Integrated analysis of long non‑coding RNA competing interactions revealed potential biomarkers in cervical cancer: Based on a public database

Cervical cancer (CC) is a common gynecological malignancy in women worldwide. Using an RNA sequencing profile from The Cancer Genome Atlas (TCGA) and the CC patient information, the aim of the present study was to identify potential long non‑coding RNA (lncRNA) biomarkers of CC using bioinformatics analysis and building a competing endogenous RNA (ceRNA) co‑expression network. Results indicated several CC‑specific lncRNAs, which were associated with CC clinical information and selected some of them for validation and evaluated their diagnostic values. Bioinformatics analysis identified 51xa0CC‑specific lncRNAs (fold‑change >2 and P<0.05), and 42 of these were included in ceRNA network consisting of lncRNA‑miRNA‑mRNA interactions. Further analyses revealed that differential expression levels of 19xa0lncRNAs were significantly associated with different clinical features (P<0.05). A total of 11xa0key lncRNAs in the ceRNA network for reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR) analysis to detect their expression levels in 31xa0pairs of CC clinical samples. The results indicated that 7xa0lncRNAs were upregulated and 4xa0lncRNAs were downregulated in CC patients. The fold‑changes between the RT‑qPCR experiments and the TCGA bioinformatics analyses were the same. Furthermore, the area under the receiver operating characteristic (ROC) curve of four lncRNAs (EMX20S, MEG3, SYS1‑DBNDD2 and MIR9‑3HG) indicated that their combined use may have a significant diagnostic value in CC (P<0.05). To the best of our knowledge, the present study is the first to have identified CC‑specific lncRNAs to construct a ceRNA network and has also provided new insights for further investigation of a lncRNA‑associated ceRNA network in CC. In additon, the verification results suggested that the method of bioinformatics analysis and screening of lncRNAs was accurate and reliable. To conclude, the use of multiple lncRNAs may thus improve diagnostic efficacy in CC. In addition, these specific lncRNAs may serve as new candidate biomarkers for clinical diagnosis, classification and prognosis of CC.

miR-93-5p Transferred by Exosomes Promotes the Proliferation of Esophageal Cancer Cells via Intercellular Communication by Targeting PTEN

OBJECTIVEnTo investigate the relationship between plasma miR-93-5p and the risk of esophageal cancer, as well as the influence of miR-93-5p on the biological function of esophageal cancer cells, exerted through exosomes.nnnMETHODSnThe expression of plasma miR-93-5p in esophageal cancer patients and healthy controls was analysed by real-time quantitative PCR. The influence of miR-93-5p on the risk and prognosis of esophageal carcinoma was analyzed by conditional logistic regression and survival analysis. The effect of miR-93-5p on the biological function of recipient cells was investigated by establishing an in vitro donor cell co-culture model. The target gene of miR-93-5p was validated by luciferase reporter assay and Western Blotting.nnnRESULTSnUpregulation of plasma miR-93-5p expression significantly increases the risk of esophageal cancer and is associated with poor prognosis. miR-93-5p transferred by exosomes promotes the proliferation of recipient esophageal cancer cells and affects the expression of PTEN and its downstream proteins p21 and cyclin D1.nnnCONCLUSIONnOur study provides a reference for the identification of biomarkers for the diagnosis and prognosis of esophageal cancer.

Aluminum Oxide Nanoparticles Upregulate ED1 Expression in Rat Olfactory Bulbs by Repeated Intranasal Instillation

Respiratory route is one of the major exposure routes to nanoparticles. The environmental agent aluminum is intensively investigated for the association with development of neurodegeneration. To evaluate potential neurotoxicity induced by aluminum oxide (Al2O3) nanoparticles, male rats were intranasally instilled with 0.1 or 1 (Al) mg/kg nanoAl2O3 or aluminum chloride (AlCl3) every two days for 60 days, using pure water as vehicle control. Neurotoxicity effects were determined by behavioural studies and immunohistochemistry staining of ED1 and beta-amyloid precursor protein (Aβ). Neither of nanoAl2O3 treated groups showed significant alterations in Morris water maze tests, however, increased escape latency were observed in 1mg/kg AlCl3 treated rats. Further, upregulation of ED1 expression were showed in olfactory bulb of 1 mg/kg nanoAl2O3 and AlCl3 exposed rats. Massive Aβ expressions were observed in whole brain of 1mg/kg (Al) AlCl3 treated rats. ED1 expression is a marker of microglia/macrophages activation, suggesting stimulus of Al2O3 nanoparticles to microglia/macrophages located in olfactory bulb and perivascular areas. In these studies, Al2O3 nanoparticles didnt show any alterations on spacial learning behaviours of rats and expression of Aβ of neuron, therefore, display lower neural effects than AlCl3.

Using Immortalized Human Lymphocytes Cell Lines to Screen the Genetic Markers of ADH5 to Formaldehyde

Formaldehyde is a carcinogen with highly toxic effect on organisms. ADH5 gene encodes the human formaldehyde dehydrogenase and is important to the detoxification of formaldehyde. The purpose of this work was to establish a platform to study ADH5 genetic markers induced by formaldehyde. After analyzing ADH5 SNPs database of Chinese Population, the three specific SNP sites : rs1154409,rs28730619,rs1154414 were sequenced in Human immortalized B lymphocytes of Chinese Han population by DNA sequencing. The real-time reverse transcription polymerase chain reaction (RT-PCR) approach was used to estimate expression of ADH5 mRNAs influenced by SNPs structure in human immortalized lymphocytes. Different immortalized B lymphocytes lines are gained by genotyping of multiple SNP sites of ADH5 gene. The mRNA expression of ADH5 could up regulate by 0.0005% formaldehyde for 48h exposure. But it showed rs1154409,rs28730619,rs1154414 SNPs has no association with the expression of ADH5 mRNA induced by formaldehyde. The human immortality lymphocytes model might be an efficient tool in a detection of genetic biomarkers of susceptibility to chemicals. The further research should be explored in the protein expression in more cell lines.