Identification of Hub Genes as Potential Prognostic Biomarkers in Cervical Cancer Using Comprehensive Bioinformatics Analysis and Validation Studies.

BACKGROUND: Cervical cancer belongs to one of the most common female cancers; yet, the exact underlying mechanisms are still elusive. Recently, microarray and sequencing technologies have been widely used for screening biomarkers and molecular mechanism discovery in cancer studies. In this study, we aimed to analyse the microarray datasets using comprehensive bioinformatics tools and identified novel biomarkers associated with the prognosis of patients with cervical cancer. METHODS: The differentially expressed genes (DEGs) from Gene Expression Omnibus (GEO) datasets including GSE138080, GSE113942 and GSE63514 were analysed using GEO2R tool. The functional enrichment analysis was performed using g:Profiler tool. The protein-protein interaction (PPI) network construction and hub genes identification were performed using the STRING database and Cytoscape software, respectively. The hub genes were subjected to expression and survival analysis in the cervical cancer. The EdU incorporation and Cell Counting Kit-8 assays were performed to evaluate the effects of hub gene knockdown on the proliferation of cervical cancer cells. RESULTS: A total of 89 overlapping DEGs (63 up-regulated and 26 down-regulated genes) were identified in the microarray datasets. The functional enrichment analysis indicated that the overlapping DEGs were mainly associated with "DNA replication" and "cell cycle". Furthermore, the PPI network analysis revealed that the network contains 87 nodes and 309 edges. Sub-module analysis using the Molecular Complex Detection tool identified 21 hub genes from the PPI network. The expression levels of the 21 hub genes were all up-regulated in the cervical cancer tissues when compared to normal cervical tissues as analysed by GEPIA tool. The survival analysis showed that the low expression of cell division cycle 45 (CDC45), GINS complex subunit 2 (GINS2), minichromosome maintenance complex component 2 (MCM2) and proliferating cell nuclear antigen (PCNA) was significantly correlated with the shorter overall survival of patients with cervical cancer. Moreover, the protein expression levels of GINS2, MCM2 and PCNA, but not CDC45, were significantly up-regulated in the cervical cancer tissues when compared to normal cervical tissues. Finally, knockdown of MCM2 significantly suppressed the proliferation of HeLa and SiHa cells. CONCLUSION: In conclusion, we screened a total of 89 overlapping DEGs from the GEO datasets, and further analysis identified four hub genes (CDC45, GINS2, MCM2 and PCNA) that were likely associated with the prognosis of patients with cervical cancer. MCM2 knockdown repressed the cervical cancer cell proliferation. The current findings may provide novel insights into understanding the pathophysiology of cervical cancer and develop therapeutic targets for patients with cervical cancer.

Integrated analysis of circRNA-miRNA-mRNA regulatory network identifies potential diagnostic biomarkers in diabetic foot ulcer.

Diabetic foot ulcer (DFU) is a common and serious complication of diabetes mellitus, which influences patients' quality of life. Recently, circRNA regulated the mRNA levels by functioning as miRNA sponge in various disease, including diabetes mellitus. Nevertheless, the circRNA-miRNA-mRNA regulatory network involved in DFU remains obscure. The aim of this study is to construct a competing endogenous RNA (ceRNA) network and screen biological indicators as diagnostic factors in DFU. All the differentially expressed circRNAs, miRNAs and mRNAs were derived from Gene Expression Omnibus database. Furthermore, circRNAs identified by cytoHubba analysis and miRNAs obtained by human miRNA-disease database were used to construct DFU-specific ceRNA network with intersection of mRNAs. Functional enrichment analysis displayed the function and pathway of dysregulated mRNAs. Hub genes with high diagnostic value were screened by ClusterONE, GO semantic similarity and receiver operating characteristic (ROC) curve. Here, the ceRNA network consisted of 8 circRNAs, 11 miRNAs and 91 mRNAs. Functional enrichment analysis demonstrated diabetic complications-related pathway including TGF-beta, FoxO and Wnt signaling pathway. GO semantic similarity and ROC curve analysis showed 6 hub genes with high diagnostic value (the area under the ROC curve ≥ 0.8) in patients with DFU, including BCL2, CCND1, IRAK4, SMAD4, SP1 and SUFU, which were identified as potential target genes for DFU diagnosis. In conclusion, the present study looked at a circRNA-miRNA-mRNA regulatory network with DFU and screened the potential function of mRNA, then identified novel diagnostic biomarkers and therapeutic targets for patients with DFU.

MiR-147b inhibits cell viability and promotes apoptosis of rat H9c2 cardiomyocytes via down-regulating KLF13 expression.

Recently, microRNAs (miRNAs) have been shown to involve in the process of heart failure. This study aims to investigate the functional role of miR-147b in rat H9c2 cardiomyocytes and explore the underlying molecular mechanisms. Cell viability of H9c2 cells was detected by MTT assay. Cell apoptosis was detected by flow cytometry. Expression of miR-147b and KLF13 mRNA was detected by quantitative real-time PCR. The relationship between miR-147b and KLF13 was verified by dual-luciferase reporter assay. Protein levels were detected by western blot analysis. It was found that H2O2 inhibited cell viability and promoted cell apoptosis of H9c2 cells in a concentration-dependent manner. MiR-147b overexpression suppressed cell viability and increased apoptosis in H9c2 cells, while knock-down of miR-147b increased cell viability and reduced apoptosis in H2O2-treated H9c2 cells. Luciferase reporter assay and in vitro functional assay showed that KLF13 was a downstream target of miR-147b, and KLF13 knock-down suppressed cell viability and induced apoptosis in H9c2 cells. Enforced expression of KLF13 restored the effects of miR-147b overexpression on cell viability and apoptosis in H9c2 cells. MiR-147b modulated the expression levels of apoptosis-related proteins, and the effects of miR-147b overexpression on apoptosis-related proteins levels were prevented by enforced expression of KLF13 in H9c2 cells. The in vivo experiments showed that miR-147b was up-regulated, and KLF13 was down-regulated in the myocardial tissues from rats with chronic heart failure. Collectively, miR-147b inhibits viability and promotes cell apoptosis by targeting KLF13 in H9c2 cells, which may be associated with the pathogenesis of heart failure.

Serological Analysis and Drug Resistance of Chlamydia pneumoniae and Mycoplasma pneumoniae in 4500 Healthy Subjects in Shenzhen, China.

OBJECTIVE: To understand the prevalence and distribution of Chlamydia pneumoniae (CP) and Mycoplasma pneumoniae (MP) in the population and to provide a basis for the prevention and treatment of respiratory tract infection. METHODS: This study included a total of 4500 healthy subjects who were given physical examination in Shenzhen People's Hospital from January to December in 2016. Venous blood was drawn from people to detect the MP- and CP-specific IgG and IgM in the serum using chemiluminescence immunoassay (CLIA). The relationship of MP and CP infections with patient age, seasons, and percentage of infections was analyzed. CONCLUSION: CP and MP cause high rate of asymptomatic infection, which may be associated with the high incidence of CP and MP infection, especially in children and the elderly population. Therefore, the implementation of effective and practical prevention measures has become an urgent need. MP culture and drug sensitivity test should be performed as early as possible in patients with manifested MP infections in order to ensure timely and proper treatment and to reduce the emergence of drug-resistant strains.

Case Report: Bullous Scabies in Two Children below 10 Years.

Bullous scabies is an infrequent and atypical presentation of scabies, with predilection for elderly and males. Its median age of presentation is 70 years. We report two male cases of bullous scabies who were 7 years and 6 months old. Both patients had excellent response to sulfur 10% ointment alone and have had no recurrence in more than 3 months of follow-up.

The correlations between system treatment efficiencies and aboveground emergent macrophyte nutrient removal for the Hsin-Hai Bridge phase II constructed wetland.

This study investigated the correlations between the system treatment efficiencies and total nitrogen (TN) and total phosphorus (TP) accumulations of aboveground tissues of the wetland macrophytes of Hsin-Hai Bridge phase II constructed wetland. Among 19 emergent macrophytes studied, the optimal TN contents, 3.82% and 3.52% (w/w) were found for water spinach (Ipomoea aquatica) and Ludwigia x taiwanensis; while the optimal TP contents were found for the above two macrophytes at 0.64% and 0.83% (w/w). The accumulations of total plant TN and TP uptakes increased from 213 to 403 kg and 41 to 75 kg from March 2007 to the peak at September 2007, respectively. The TN ratios between plant tissue accumulations and the removals from the influents were 1.57%, 2.76%, 1.51% and 3.2% from March 2007 to March 2008. In the same period, the TP ratios between plant tissue accumulations and the removals from influents were 1.71%, 8.0%, 0.58% and 10.1%. The roles of the uptakes by aboveground portions of emergent macrophytes in system nutrient removals from the influents were more significant during growth seasons.

Evaluation of the polymorphic D-loop of Columba livia in forensic applications.

We report on the polymorphisms exhibited by three hypervariable regions within the D-loop of Columba livia (pigeon) mitochondrial DNA. A total of 131 samples were taken from 131 randomly selected birds and used in the analyses of SNPs, a variable number of tandem repeats (VNTR) and an STR locus using CE. The number of repeats for the VNTR ranged from 2 to 8 producing 21 haplotypes, with 54 individuals exhibiting heteroplasmy. The STR locus exhibited multiple and continuous repeats within each individual and these patterns were not reproducible with individuals of the same maternal lineage, where different haplotypes were noted. Combining the SNP and VNTR loci produced 38 haplotypes, with the power of discrimination being 0.93. The polymorphic regions of D-loop observed in this study are potential markers for maternal relationship identification.

Establishing the mitochondrial DNA D-loop structure of Columba livia.

The complete mitochondrial DNA D-loop structure of pigeon (Columba livia) was established in this study. A strategy of amplifying three partial fragments of the D-loop and then combing the three fragments to cover the full length of the D-loop was adopted. Ten samples from pigeons were collected and were successfully amplified and sequenced. Repetitive sequences of a VNTR and an STR were both observed at the 3'-end of D-loop region. DNA sequence data revealed polymorphic sequences including indels, SNP, VNTR and STR within the D-loop. The size of the D-loop ranged from 1310 to 1327 bp from the initiation site of D-loop to the site immediately upstream of the repeat sequences depending upon the number of insertions or deletions. Each sample could be distinguished based on four genotyping procedures; being indels, SNPs, VNTRs and STRs. The polymorphic nature of the D-loop can be a valuable method for maternal identification and genetic linkage of pigeon in particular forensic science investigations.

Species identification using the cytochrome b gene of commercial turtle shells.

Turtle shells and their gelled products are familiar in some countries as foods, tonics and medicines. These shells may come from endangered and protected species, requiring the identification of the species present to enforce national and international legislation. We report on the design of five combinations of primer pairs for the identification of turtle shells and shell fragments used as ornaments, food products and medicines. The types of samples used are those encountered frequently and will typically contain highly degraded DNA. The success rate for species identification using the test described is dependent upon the choice of primer sets used and the length of the expected amplification product. Gelled products were simulated by the process of decoction for up to 12 h, after which all the turtle species could be identified from the liquid samples. This study establishes a method for the identification of commercial turtle shells and illustrates a simulated case using gelled products.