Construction of the Six-lncRNA Prognosis Signature as a Novel Biomarker in Esophageal Squamous Cell Carcinoma.

Esophageal squamous cell carcinoma (ESCC) is a common malignant gastrointestinal tumor threatening global human health. For patients diagnosed with ESCC, determining the prognosis is a huge challenge. Due to their important role in tumor progression, long non-coding RNAs (lncRNAs) may be putative molecular candidates in the survival prediction of ESCC patients. Here, we obtained three datasets of ESCC lncRNA expression profiles (GSE53624, GSE53622, and GSE53625) from the Gene Expression Omnibus (GEO) database. The method of statistics and machine learning including survival analysis and LASSO regression analysis were applied. We identified a six-lncRNA signature composed of AL445524.1, AC109439.2, LINC01273, AC015922.3, LINC00547, and PSPC1-AS2. Kaplan-Meier and Cox analyses were conducted, and the prognostic ability and predictive independence of the lncRNA signature were found in three ESCC datasets. In the entire set, time-dependent ROC curve analysis showed that the prediction accuracy of the lncRNA signature was remarkably greater than that of TNM stage. ROC and stratified analysis indicated that the combination of six-lncRNA signature with the TNM stage has the highest accuracy in subgrouping ESCC patients. Furthermore, experiments subsequently confirmed that one of the lncRNAs LINC01273 may play an oncogenic role in ESCC. This study suggested the six-lncRNA signature could be a valuable survival predictor for patients with ESCC and have potential to be an auxiliary biomarker of TNM stage to subdivide ESCC patients more accurately, which has important clinical significance.

Is Human-induced Pluripotent Stem Cell the Best Optimal?

OBJECTIVE: Since the advent of induced pluripotent stem cell (iPSC) technology a decade ago, enormous progress has been made in stem cell biology and regenerative medicine. Human iPSCs have been widely used for disease modeling, drug discovery, and cell therapy development. In this review, we discuss the progress in applications of iPSC technology that are particularly relevant to drug discovery and regenerative medicine, and consider the remaining challenges and the emerging opportunities in the field. DATA SOURCES: Articles in this review were searched from PubMed database from January 2014 to December 2017. STUDY SELECTION: Original articles about iPSCs and cardiovascular diseases were included and analyzed. RESULTS: iPSC holds great promises for human disease modeling, drug discovery, and stem cell-based therapy, and this potential is only beginning to be realized. However, several important issues remain to be addressed. CONCLUSIONS: The recent availability of human cardiomyocytes derived from iPSCs opens new opportunities to build in vitro models of cardiac disease, screening for new drugs and patient-specific cardiac therapy.

SOD mimetic improves the function, growth, and survival of small-size liver grafts after transplantation in rats.

BACKGROUND: Small-for-size syndrome (SFSS) may occur when graft volume is less than 45% of the standard liver volume, and it manifests as retarded growth and failure of the grafts and more mortality. However, its pathogenesis is poorly understood, and few effective interventions have been attempted. AIMS: The present study aimed to delineate the critical role of oxidant stress in SFSS and protective effects of a superoxide dismutase mimetic, Mn(III)tetrakis(4-benzoic acid)porphyrin chloride (MnTBAP), on graft function, growth, and survival in the recipient rats. METHODS: Small size graft liver transplantation (SSGLT) was performed to determine the survival, graft injury, and growth. MnTBAP was administered in SSGLT recipients (SSGLT+MnTBAP). RESULTS: Serum alanine aminotransferase levels were sustained higher in SSGLT recipients, which were correlated with an increased apoptotic cell count and hepatocellular necrosis in liver sections. Malondialdehyde content, gene expression of tumor necrosis factor α and interleukin 1ß, and DNA binding activity of nuclear factor-κB in the grafts were increased significantly in SSGLT recipients compared with sham-operated controls. Both phosphorylated p38 mitogen-activated protein kinase and nuclear c-Jun were increased in SSGLT. All these changes were strikingly reversed by the administration of MnTBAP, with an increase in serum superoxide dismutase activity. Moreover, in situ bromodeoxyuridine incorporation demonstrated that graft regeneration was much more profound in the SSGLT+MnTBAP group than in the SSGLT group. Finally, the survival of recipients with MnTBAP treatments was significantly improved. CONCLUSIONS: Enhanced oxidant stress with activation of the p38/c-Jun/nuclear factor-κB signaling pathway contributes to SFSS-associated graft failure, retarded graft growth, and poor survival. MnTBAP effectively reversed the pathologic changes in SFSS-associated graft failure.

miRNA regulation of liver growth after 50% partial hepatectomy and small size grafts in rats.

BACKGROUND: The molecular mechanisms underlying the growth of small size grafts and the remaining livers are poorly understood. MicroRNAs (miRNAs) negatively modulate expression of genes that are involved in cellular function and metabolism. The aim of this study is to identify critical miRNA species that modulate the growth of small grafts and the remaining livers after partial hepatectomy (PH). METHODS: Small size graft liver transplantation was performed in rats. Liver tissue was harvested after transplant or PH for the determination of miRNA expression profile, and the data were confirmed by quantitative reverse-transcriptase polymerase chain reaction. The genes involved in cell cycle and proliferation were analyzed by quantitative reverse-transcriptase polymerase chain reaction and immunohistochemical staining. RESULTS: Compared with control liver, miR_122a, Let_7b, and miR_26a were reduced by more than 90% in 45% volume grafts. In the remaining livers after 50% PH, 30 miRNAs were down-regulated by more than 50%, and among them, miR_22a, miR_26a, miR_30b, Let_7f, and Let_7g were markedly decreased. A negative correlation existed between down-regulated miRNAs and highly up-regulated genes involved in cell cycle and proliferation in the remaining livers. Moreover, overexpression of miR_26a markedly down-regulated cyclin E2 protein levels and significantly decreased proliferation of HepG2 cells. CONCLUSION: Down-regulated miRNAs play a pivotal role in promoting the growth of small size grafts and the remaining livers. The negative correlation between down-regulated miRNAs and up-regulated genes suggests that these specific miRNAs participate in the modulation of a growth response in both living donors and small size graft recipients.

Immunohistochemical staining of cancer stem cell markers in hepatocellular carcinoma.

BACKGROUND: Cancer stem cells (CSCs) are thought to be a critical subpopulation in tumor development, progression, metastasis and recurrence, and the identification of these cells is an initial step in understanding their role in oncogenesis and in seeking valuable markers for diagnosis or development of targeting therapeutics. AIMS: To identify CSCs in hepatocellular carcinoma (HCC) specimens and define their tissue specificity. METHODS: Immunohistochemical staining of CSC markers: CD44, CD90, CD133 and aldehyde dehydrogenase (ALDH) was performed in 25 HCC specimens, 4 hepatoblastomas, 8 peri-malignant tissues, and 19 cases of viral hepatitis. RESULTS: The positivity of CD44 staining in HCC specimens was significantly lower than in viral hepatitis specimens. The positive rate of CD133 in HCC was similar to viral hepatitis specimens. CD133(+) cells were largely localized to ALDH-positive cells in HCC as revealed by confocal microscopy. In contrast, the co-expression of both markers was visualized within vessels or in the portal areas in viral hepatitis. Moreover, among 7 liver specimens adjacent to HCC tissue, 3-6 samples were positive for CD44, CD90, CD133 and ALDH, especially in dysplastic cells. One of 4 hepatoblastoma cases was positive for all these markers; whereas, the other three specimens were negative for all these CSC markers. CONCLUSIONS: In HCC and dysplastic tissues, clusters of CD133(+)/ALDH(high) cells were identified. The use of cancer stem cell markers to screen tissues with chronic liver diseases provides limited guidance in the identification of malignant cells.

Physico-chemical characterization of polylipid nanoparticles for gene delivery to the liver.

Polylipid nanoparticles (PLNP) have been shown to be very effective in delivering antioxidative genes in the treatment of liver injury in mice. To build on our previous studies and to further characterize PLNP formulated from polycationic lipid (PCL) and cholesterol, we report here the synthesis of multigram quantities of PCL and employ analytical tools, such as Raman spectroscopy of single PLNP and live-cell imaging of lipofection, for the physicochemical characterization of PCL, PLNP, and the transfection process. Mass spectrometry demonstrates the characteristics of polymeric lipids. Raman spectrum of PCL reveals the polymeric structure of the polymers. The presence of cholesterol in PLNP formulation did not markedly change the Raman spectrum. PLNP-derived polyplexes exhibit Raman spectra very similar to PLNP except that the C-H out-of-plane deformation mode of the polymeric lipid is significantly suppressed, indicating the interaction with plasmid DNA. Zeta potential measurement indicates a large DNA-carrying capacity of PLNP and their stability for in vivo gene delivery. The live-cell fluorescent imaging dynamically shows that PLNP exerts transfection efficiency similar to lipofectamine in leading to early reporter gene expression in live hepatic cells. In conclusion, polylipid nanoparticles possess a high DNA carrying capacity and lipofection efficiency, rendering them suitable for testing in large animals. The employment of novel state-of-the-art technologies in the study of lipofection represents the level of physicochemical and biological characterization that is needed to best understand the key elements involved in the lipofection process.

[Implication of different expression of IL-2 mRNA and IL-10 mRNA in CD4(+)CD25(+)T cell induced immune tolerance of liver transplantation in rat].

OBJECTIVE: To investigate the expression of tolerance-associated interleukin (IL)-2 mRNA and IL-10 mRNA in rats after allogeneic liver transplantation. METHODS: The experimental rats were randomly divided into 3 groups: acute rejection model group (group I), CD4(+)CD25(+)T cell treatment group (group II), donors of the two groups were Wistar while recipients were Sprague-Dawley (SD) rats, and group III as control group, both donors and recipients of this group were SD rats, with 12 rats in each group. Splenic lymphocyte of donor in group II were injected through vena dorsalis penis 7 days before liver transplantation; equal volume normal saline (NS) were injected in rats of group I and III. The contents of IL-2 mRNA and IL-10 mRNA in the liver tissue were determined by reverse transcription-polymerase chain reaction (RT-PCR), and the number of lymphocytes in the donor liver was quantified with flow cytometry 7 days after transplantation. Meanwhile the pathologic change in the donor tissue were observed, and the recipients' life span was recorded. RESULTS: IL-2 mRNA was expressed strongly in the liver of group I, but it was expressed weakly in group II, and no expression was detected in the liver of group III. IL-10 mRNA was expressed only in group II, and the level was high. The rats in group II and III survived over 30 days, but rats in group I had a shorter life (8-11 days, both P<0.01). There was heavy lymphocytic infiltration in the liver of group I,and was much higher than that of the other groups [group I:(14.31+/-3.41)x10(6)/g, group II: (5.04+/-1.13) x10(6)/g, group III: (1.55+/-0.40) x10(6)/g, all P<0.01], and pathology showed moderate rejection. Rats in group II had milder lymphocytic infiltration, and pathology showed no signs of rejection or uncertain rejection, and the ratio of CD4(+) T cell and CD4(+)CD25(+) were higher than those of group I [(43.31+/-8.07)% vs. (33.65+/-7.25)% and (11.39+/-1.92)% vs. (3.05+/-0.62)%] and group III [(43.31+/-8.07)% vs. (31.18+/-6.25)% and (11.39+/-1.92)% vs. (3.37+/-0.72)%, P<0.05 or P<0.01]. In group III no lymphocytic infiltration was found, and pathology showed no sign of rejection. CONCLUSION: IL-2 may participate in the immune rejection in allogeneic liver transplantation, but IL-10 plays an important role in CD4(+)CD25(+) T cell inducing immune tolerance of allogeneic liver transplantation in the rats.