Construction of molecular subgroups of ulcerative colitis.

OBJECTIVE: Ulcerative colitis (UC), a chronic inflammatory disease of the colon with unknown etiology, is characterized by remission and recurrence. At present, a considerable number of UC cases are misdiagnosed or delayed in diagnosis and treatment. We aimed to identify UC-related genes to aid the development of drugs for this condition. PATIENTS AND METHODS: Transcriptome data of 362 patients with UC and 126 control subjects were obtained from the Gene Expression Omnibus. The 362 patients with UC were subgrouped using unsupervised machine learning. R software was used to analyze the clinical characteristics of the subgroups, screen subgroup-specific genes, assess the relationships between gene modules and clinical characteristics using weighted gene co-expression network analysis, and perform Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses of the subgroups. RESULTS: Patients with UC were classified into two subgroups. Genes specific to subgroup I included IL21R, ATP8B2, and PLEKHO1. Severe disease tended to be associated with immune cell infiltration; anti-tumor necrosis factor (TNF)-α antibodies and ustekinumab may have been effective in this subgroup. Subgroup II-specific genes included SLC4A4, EPB41L4B, and PLCE1. Patients in this subgroup had mild clinical conditions; however, their disease was more likely to progress to colorectal cancer. Thus, 5-aminosalicylic acid-based drugs may be effective for the treatment of UC in these patients. CONCLUSIONS: We divided UC into two molecular subgroups based on transcriptome data, providing molecular evidence for the development of diagnostic methods and individualized treatment strategies for UC.

A stress-induced cellular aging model with postnatal neural stem cells.

This corrects the article DOI: 10.1038/cddis.2014.82.

Analysis of circulating long non-coding RNA UCA1 as potential biomarkers for diagnosis and prognosis of osteosarcoma.

OBJECTIVE: The aim of this investigation was to examine the potential usefulness of long non-coding RNA UCA1 (UCA1) as a biomarker for diagnosis and prognosis in osteosarcoma. PATIENTS AND METHODS: The expression level of UCA1 was determined using TaqMan real-time PCR in human osteosarcoma tissues and patients' sera. Next, we investigated to clarify the relationship of UCA1 with clinicopathological features. The receiver operating characteristic (ROC) curve was performed to estimate the diagnostic value of UCA1. Finally, the prognosis of patients with osteosarcoma was assessed by Kaplan-Meier method and Cox proportional hazards model. RESULTS: We observed that UCA1 was significantly increased in osteosarcoma tissue compared with normal bone tissue (p<0.001) and the serum expression of UCA1 was significantly higher in patients with osteosarcoma than that in healthy controls (p<0.01). Up-regulation of UCA1 was correlated with clinical stage (p=0.001) and metastasis (p=0.007). Furthermore, serum UCA1 levels were observed to be robust in differentiating osteosarcoma patients from control subjects [area under the curve (AUC) = 0.831; 95% confidence interval (CI)= 0.746 to 0.916]. Kaplan-Meier analysis showed that that high UCA1 expression level was associated with poorer overall survival (p<0.001) and disease-free survival (p<0.001). Finally, Cox regression analyses showed that UCA1 expression might be an independent prognostic parameter to predict poor prognosis. CONCLUSIONS: Our study firstly showed that UCA1 could be a specific and noninvasive candidate biomarker for the diagnosis and prognosis of UCA1.

miR-542-3p targets sphingosine-1-phosphate receptor 1 and regulates cell proliferation and invasion of breast cancer cells.

OBJECTIVE: MicroRNAs (miRs) regulate the proliferation and metastasis of numerous cancer cell types. This study aimed to reveal the role of microRNA-542-3p (miR-542-3p) in breast cancer (BC) cell proliferation and its potential mechanisms. MATERIALS AND METHODS: MiR-542-3p expression was detected by reverse transcription-polymerase chain reaction (RT-PCR) and sphingosine-1-phosphate receptor 1(S1PR1) protein expression was measured by Western blotting. TargetScan was used to predict the downstream target genes of miR-542-3p, which were confirmed by luciferase and RNA immunoprecipitation assays. Biological functions of miR-542-3p and S1PR1 were analyzed using CKK-8, colony formation, migration, and invasion. RESULTS: It was demonstrated that the expression of miR-542-3p was downregulated in BC tissues and cell lines. We also showed that the expression of S1PR1 was upregulated in BC tissues and cell lines. Furthermore, we found that the expression level of miR-542-3p was negatively correlated with the expression level of S1PR1 in BC tissues. Over-expression of miR-542-3p inhibited BC cell proliferation, colony formation, migration and invasion. The dual luciferase reporter experiments showed that miR-542-3p regulated the expression of S1PR1 by combining with its 3'UTR. Finally, we showed that down-expression of miR-542-3p inhibited BC cell proliferation, colony formation, migration and invasion. CONCLUSIONS: Our study provides the new insight that miR-542-3p inhibited colon cancer cells via targeting S1PR1, suggesting that miR-542-3p/S1PR1 can serve as a potential therapeutic target for BC.

PET/CT imaging in diagnosing lymph node metastasis of esophageal carcinoma and its comparison with pathological findings.

OBJECTIVE: To investigate the usefulness of 18F-FDG PET/CT imaging in the diagnosis of lymph node metastasis of esophageal carcinoma and to compare its results with pathological findings. PATIENTS AND METHODS: This study examined 43 cases of patients (32 males and 11 females, aged 54 ± 13 years old) being diagnosed with lymph node metastasis of esophageal carcinoma in our hospital between 2005 and 2014. All of these patients accepted 18F-FDG PET/CT imaging 10 days before the operation. Before reconstruction, each patient went through the body scan. PET/CT images were subject to comprehensive diagnostic analyses, by three experienced radiologists and/or professional nuclear physicians, on the number of metastatic lymph nodes and the maximum standardized uptake value (SUVmax). A control study was also performed on the pathological findings according to the latest esophageal cancer lymph node partition. RESULTS: A total of 846 lymph nodes were taken out from the patients, among which 154 were confirmed with metastasis. When the SUVmax cutoff values were set at 2.5 and 5, 201 and 173 metastatic lymph nodes were found by 18F-FDG PET/CT imaging, respectively. Additionally, under such condition, 18F-FDG PET/CT imaging had the esophageal sensitivity of 69.48% vs 87.66%, specificity of 92.71% vs 94.51%, accuracy of 83.33% vs 93.26%, positive predictive value of 53.23% vs 78.03%, and negative predictive value of 92.71% vs 97.18%. CONCLUSIONS: 18F-FDG PET/CT had high accuracy in imaging lymph node metastasis of esophageal cancer. SUVmax cutoff value of 5 had a higher diagnostic accuracy and should be recommended in clinical practice.

Identification of key genes involved in HER2-positive breast cancer.

OBJECTIVE: As an invasive cancer, breast cancer is the most common tumour in women and is with high mortality. To study the mechanisms of HER2-positive breast cancer, we analyzed microarray of GSE52194. MATERIALS AND METHODS: GSE52194 was downloaded from Gene Expression Omnibus including 5 HER2-positive breast cancer samples and 3 normal breast samples. Using cuffdiff software, differentially expressed genes (DEGs) and differentially expressed long non-coding RNAs (DE-lncRNAs) were screened. Functions of the DEGs were analyzed by Gene Ontology (GO) and pathway enrichment analyses. Then, protein-protein interaction (PPI) network of the DEGs was constructed using Cytoscape and modules of the PPI network were screened by CFinder. Moreover, lncRNA-DEG pairs were screened. RESULTS: Total 209 lncRNA transcriptions were predicted, and 996 differentially expressed transcriptions were screened. Besides, FOS had interaction relationships with EGR1 and SOD2 separately in module E and F of the PPI network for the DEGs. Moreover, there were many lncRNA-DEG pairs (e.g. TCONS_00003876-EGR1, TCONS_00003876-FOS, lnc-HOXC4-3:1-FOS, lnc-HOXC4-3:1-BCL6B, lnc-TEAD4-1:1-FOS and lnc-TEAD4-1:1-BCL6B), meanwhile, co-expressed DEGs of TCONS_00003876, lnc-HOXC4-3:1 and lnc-TEAD4-1:1 were enriched in p53 signaling pathway, MAPK signaling pathway and cancer-related pathways, respectively. CONCLUSIONS: ANXA1, EGR1, BCL6, SOD2, FOS, TCONS_00003876, lnc-HOXC4-3:1 and lnc-TEAD4-1:1 might play a role in HER2-positive breast cancer.

A stress-induced cellular aging model with postnatal neural stem cells.

Aging refers to the physical and functional decline of the tissues over time that often leads to age-related degenerative diseases. Accumulating evidence implicates that the senescence of neural stem cells (NSCs) is of paramount importance to the aging of central neural system (CNS). However, exploration of the underlying molecular mechanisms has been hindered by the lack of proper aging models to allow the mechanistic examination within a reasonable time window. In the present study, we have utilized a hydroxyurea (HU) treatment protocol and effectively induced postnatal subventricle NSCs to undergo cellular senescence as determined by augmented senescence-associated-ß-galactosidase (SA-ß-gal) staining, decreased proliferation and differentiation capacity, increased G0/G1 cell cycle arrest, elevated reactive oxygen species (ROS) level and diminished apoptosis. These phenotypic changes were accompanied by a significant increase in p16, p21 and p53 expression, as well as a decreased expression of key proteins in various DNA repair pathways such as xrcc2, xrcc3 and ku70. Further proteomic analysis suggests that multiple pathways are involved in the HU-induced NSC senescence, including genes related to DNA damage and repair, mitochondrial dysfunction and the increase of ROS level. Intriguingly, compensatory mechanisms may have also been initiated to interfere with apoptotic signaling pathways and to minimize the cell death by downregulating Bcl2-associated X protein (BAX) expression. Taken together, we have successfully established a cellular model that will be of broad utilities to the molecular exploration of NSC senescence and aging.

Use of a new type of trocar for the surgical treatment of hydrocephalus: a simple and effective technique.

This study compared the use of a new type of peritoneocentesis trocar with conventional laparotomy for the placement of the distal catheter in the treatment of hydrocephalus with ventriculoperitoneal shunt. A total of 376 patients with hydrocephalus were recruited to the study and were assigned randomly to undergo insertion of the distal catheter by conventional laparotomy (n = 195) or using the new peritoneal trocar (n = 181). The time taken for the surgical procedure and the complication rate over the following 1-year period were compared between the two groups. The mean length of the procedure to place the distal catheter was significantly shorter in the trocar group compared with the laparotomy group. Infection and obstruction rates were significantly higher in the laparotomy group than in the trocar group. In conclusion, the use of the new trocar was associated with lower rates of surgically induced trauma and complications compared with conventional laparotomy.

Prevention of neuropathic pain in an animal model of spare nerve injury following oral immunization with recombinant adenovirus serotype 5-mediated NR2B gene transfer.

Spinal N-methyl-D-aspartate receptor 2B subunit (NR2B)-increased expression plays an important role in the facilitation and maintenance of the persistent pain state due to peripheral nerve injury. A vaccination strategy to reduce the expression of brain protein is feasible and may have therapeutic potential for neurological disorders. Thus, we investigated the effect of oral immunization with recombinant adenovirus serotype 5-mediated NR2B gene transfer (rAd5/NR2B) for the modulation of neuropathic pain. After peroral administration of the rAd5/NR2B vaccine, transgene NR2B expression persisted for at least a week and was associated with the induction of high serum titers of NR2B-specific antibodies. Following the occurrence of mechanical allodynia due to peripheral nerve injury, NR2B-specific antibodies could pass the blood-brain barrier, transport and subsequently bind to the spinal NR2B protein. The humoral immunoresponse results in the strong antiallodynia in the spared nerve injury animal model. These data proved the feasibility of oral immunization with rAd5/NR2B for the prevention of neuropathic pain.

CTLA4-Fas ligand gene transfer mediated by adenovirus induce long-time survival of murine cardiac allografts.

BACKGROUND: Fas ligand gene transfer to induce peripheral allograft tolerance in animal models has shown controversial results. The immunosuppression effects mediated by engineered FasL depend on whether alloreactive T cells are selectively deleted. In the present study, we tested the feasibility of a strategy to induce long-time survival by fusing CTLA4-FasL gene transfer in vivo. METHODS: Cardiac allografts from DA(RT-1(a)) rats were transplanted heterotopically into the abdomens of LEW(RT-1(1)) rats. Plaque units (5x10(9)) of either AdCTLA4-FasL, AdCTLA4Ig, or AdEGFP were administered via the portal vein immediately after cardiac transplantation. The frequencies of helper T lymphocyte precursors (HTLp) and cytotoxic T lymphocyte precursors (CTLp) were determined by a combined single limiting dilution assay on days 5 and 20 after transplantation. RESULTS: Cardiac allograft survival was significantly prolonged by either AdCTLA4-FasL or AdCTLA4Ig treatment(mean survival times [MST] of 71.0 +/- 3.7 and 45.7 +/- 2.4, respectively, n = 6) compared with untreated hosts or animals treated with AdEGFP(MST of 5.7 +/- 0.5 and 5.2 +/- 0.4, respectively, n = 6). In addition, treatment with AdCTLA4-FasL led to significantly prolonged allograft survival compared with AdCTLA4Ig treatment. Furthermore, the frequencies of HTLp and CTLp on day 20 among rats treated with AdCTLA4-FasL was lower than those on day 5, whereas frequencies of HTLp and CTLp on day 20 were similar with those on day 5 in the other groups. CONCLUSION: These results suggest that administration of an adenovirus encoding fusion CTLA4-FasL gene to rat recipients effectively decreased the size of alloreactive T cells and induced long-term survival of cardiac allografts.