[Effect of inhibiting miR-204 expression on the learning and memory abilities of neonatal rats with intrauterine growth restriction and related mechanism]. / 抑制miR-204è¡¨è¾¾å¯¹å®«å† å‘è‚²è¿Ÿç¼“æ–°ç”Ÿå¤§é¼ å­¦ä¹ è®°å¿†èƒ½åŠ›çš„å½±å“åŠç›¸å ³æœºåˆ¶.

OBJECTIVES: To investigate the effect of inhibiting miR-204 expression on the learning and memory abilities of neonatal rats with intrauterine growth restriction (IUGR) and related mechanism. METHODS: A rat model of IUGR was prepared by low-protein diet. The 3-day-old IUGR rats were divided into three groups: model, miRNA antagonist control and miR-204 antagonist, with 10 rats in each group. Ten normal neonatal rats served as the control group. Morris water maze test was used to measure the learning and memory abilities of the rats. Quantitative real-time PCR was used to measure the mRNA expression levels of miR-204 and brain-derived neurotrophic factor (BDNF) in the hippocampus. Nissl staining and TUNEL staining were used to observe the number of Nissl bodies and the apoptosis of cells in the hippocampus. Western blot was used to measure the expression levels of BDNF/TrkB signaling pathway-related proteins in the hippocampus. RESULTS: Compared with the control group, the model group had a significant increase in the escape latency and a significant reduction in the number of platform crossings (P<0.001). The model group also had significant increases in the apoptosis rate of cells and the expression level of miR-204 in hippocampal tissue (P<0.001), while the number of Nissl bodies, the mRNA expression level of BDNF, and the protein expression levels of BDNF, p-TrkB, and p-CREB in the model group were significantly reduced compared with the control group (P<0.001). After inhibition of the expression of miR-204, the number of Nissl bodies, the mRNA expression level of BDNF, and the protein expression levels of BDNF, p-TrkB, and p-CREB significantly increased, while the cell apoptosis rate and the expression level of miR-204 in the hippocampus significantly decreased. The escape latency was also reduced, while the number of platform crossings increased after inhibition of the expression of miR-204 (P<0.001). CONCLUSIONS: Inhibiting miR-204 can improve the learning and memory functions of neonatal rats with IUGR, possibly by targeted activation of the BDNF/TrkB signaling pathway.

LncRNA FGD5-AS1 enhances the proliferation and stemness of hepatocellular carcinoma cells through targeting miR-223 and regulating the expression of ECT2 and FAT1.

AIM: Hepatocellular carcinoma (HCC) is common and causes many deaths worldwide. The aim of this study is to explore the mechanism by which long non-coding RNA FGD5-AS1 regulates HCC cell proliferation and stemness. METHODS: Tumor and normal adjacent tissues were harvested from HCC patients. Real-time quantitative reverse transcription-PCR was applied to examine the expression of FGD5-AS1, miR-223, Epithelial cell transforming sequence 2 (ECT2) and FAT1. The protein levels of ECT2, FAT1, proliferating cell nuclear antigen (PCNA), OCT4, CD133 and CD90 were analyzed by western blot. The localization of FGD5-AS1 was examined by Fluorescence in situ hybridization. Cell proliferation was analyzed with CCK-8 and colony formation assays. Spheroid formation was used for analyzing cell stemness. Gene interaction was examined by RNA immunoprecipitation and luciferase activity assays. A subcutaneous xenograft mouse model was established to analyze HCC growth and stemness in vivo. Immunohistochemistry staining was used to analyze the expression PCNA and OCT4 in subcutaneous tumors. RESULTS: FGD5-AS1 was upregulated in HCC and its high expression indicated poor prognosis of patients. High expression of FGD5-AS1 enhanced HCC cell proliferation and stemness. Knockdown of FGD5-AS1 restrained tumor growth and stemness in mice. FGD5-AS1 directly sponged miR-223 and promoted the expression of ECT2 and FAT1 in HCC. Both knockdown of miR-223 and overexpression of ECT2 and FAT1 reversed FGD5-AS1 silencing-mediated suppression of HCC cell proliferation and stemness. CONCLUSION: FGD5-AS1 directly sponged miR-223 and promoted the expression of ECT2 and FAT1 in HCC, thus enhancing HCC cell proliferation and stemness. Our study identifies potential prognostic biomarkers and therapeutic targets for HCC.

Prognostic Value of Postoperative Circulating Tumor DNA in Patients With Early- and Intermediate-Stage Hepatocellular Carcinoma.

Majority of patients with resected early- and intermediate-stage liver cancer will experience postoperative recurrence. This study aimed to investigate the application of ctDNA sequencing in the postoperative period of hepatocellular carcinoma. A total of 96 patients with liver cancer were enrolled in this study. Postoperative peripheral blood samples were collected from all patients after surgery and analyzed using hybridization capture-based next-generation sequencing. Identification of at least one somatic mutation in the peripheral blood was defined as ctDNA+. Five genetic features in tumor tissues were associated with disease-free survival (DFS) using Lasso-Cox model. The area under the receiver operating characteristic curve was 0.813 and 0.882 in training and validation cohorts, respectively. The recurrence rate in ctDNA+ and ctDNA- groups was 60.9% and 27.8%, respectively. Multivariate Cox regression analysis showed that the postoperative ctDNA was an independent prognostic predictor of DFS (HR [hazard ratio]: 6.074, 95% Cl [confidence interval]: 2.648-13.929, P<0.001) and overall survival (OS) (HR: 4.829, 95% CI: 1.508-15.466, P=0.008). Combined ctDNA with AFP improved prediction performance. The median DFS was 2.0, and 8.0 months in ctDNA+/AFP-H and ctDNA+/AFP-L groups, respectively; while ctDNA-/AFP-H and ctDNA-/AFP-L groups had not reached the median time statistically (Log-rank test, P < 0.0001). Furthermore, ctDNA- patients had better prognosis than ctDNA+ patients irrespective of tumor stage. Postoperative ctDNA sequencing has great prognostic value in patients with liver cancer. Patients with positive ctDNA should receive more intensive disease monitoring and more aggressive treatment strategies to improve the survival time.

N,S,P Co-Doped Carbon Nanodot Fabricated from Waste Microorganism and Its Application for Label-Free Recognition of Manganese(VII) and l-Ascorbic Acid and AND Logic Gate Operation.

A novel fluorescent probe based on N,S,P codoped carbon nanodots (N,S,P-CNDSac) is very simple and quickly fabricated by a one-step hydrothermal pyrolysis of Saccharomyces cerevisiae and utilized for label-free and "on-off-on" sequential detection of manganese(VII) and l-ascorbic acid (l-AA). The fluorescence of N,S,P-CNDSac can be effectively quenched by Mn(VII) based on an inner filter effect (IFE) and recovered upon the addition of l-AA due to the easy conversion of Mn(VII) to reduced states (i.e., Mn(IV), Mn(II), and Mn(0)) by l-AA. This probe exhibited favorable selectivity and sensitivity toward Mn(VII) and l-AA with detection limits of 50 nmol/L and 1.2 µmol/L, respectively. Simultaneously, an "AND" logic gate based on the as-fabricated N,S,P-CNDSac has been constructed. Also, the as-proposed fluorescent probe was extended to detect Mn(VII) and l-AA in biosystems. Furthermore, the as-constructed fluorescent probe system was successfully applied to the analyses of Mn(VII) in tap water, Fenhe River water, and medicinal herb samples with satisfactory results. The proposed method is simple and easily accessible, demonstrating the great potential of N,S,P-CNDSac in biosensing, disease diagnosis, cellular labeling, and environmental monitoring.

[Value of postoperative indocyanine green retention rate at 15 minutes combined with standard remnant liver volume in predicting liver dysfunction after hepatectomy].

OBJECTIVE: To investigate the value of indocyanine green retention rate at 15 minutes (ICG R15) on postoperative day 3 combined with standard remnant liver volume (SRLV) in predicting the occurrence of liver dysfunction after hepatectomyin hepatocellular carcinoma (HCC).
 Methods: The clinical data of 61 HCC patients undergone hepatectomy in Xiangya Hospital of Central South University from January 2015 to February 2016 were collected and analyzed. The patients were divided into 2 groups: a normal liver function group (n=40) and a liver dysfunction group (n=21). Univariate analysis was used to evaluate the risk factors for postoperative liver dysfunction. Logistic regression was used to assess the independent risk factors for postoperative liver dysfunction, and the regression equation between independent risk factors and postoperative liver dysfunction was established. The receiver operating characteristic (ROC) curve was used to examine the regression equation and compare the value difference in predicting postoperative liver dysfunction between single and combined independent risk factors.
 Results: Postoperative liver dysfunction occurred in 21 of the 61 patients, with an incidence rate at 34.4%. There was no significant difference in the time of operation, time of hepatic portal occlusion, volume of tumor and volume of resected liver between the 2 groups (all P>0.05), but there were significant differences in the ICG R15 on postoperative day 3, intraoperative blood loss and SRLV between the 2 groups (all P<0.05). The ICG R15 on postoperative day 3, intraoperative blood loss, SRLV were the risk factors for postoperative liver dysfunction. Logistic regression analysis showed ICG R15 on postoperative day 3 and SRLV were the independent risk factors for postoperative liver dysfunction, and the regression equation between independent risk factors and postoperative liver dysfunction was as follows: logit(P)=1.277+0.140×ICG R15 on postoperative day 3-5.125×SRLV. The area under the ROC curve of ICG R15 on postoperative day 3 combined with SRLV was more than that of single ICG R15 and single SRLV.
 Conclusion: ICG R15 on postoperative day 3 and SRLV are the independent risk factors for postoperative liver dysfunction. The regression equation, which is established by combination of ICG R15 with SRLV, can predict the occurrence of postoperative liver dysfunction. The accuracy of ICG R15 on postoperative day 3 combined with SRLV is better than that of single ICG R15 or single SRLV.

Two-liquid-phase system: A promising technique for predicting bioavailability of polycyclic aromatic hydrocarbons in long-term contaminated soils.

A two-liquid-phase system (TLPS), which consisted of soil slurry and silicone oil, was employed to extract polycyclic aromatic hydrocarbons (PAHs) in four long-term contaminated soils in order to assess the bioavailability of PAHs. Extraction kinetics of six PAHs viz. phenanthrene, fluoranthene, pyrene, benzo(a)anthracene, benzo(a)pyrene, dibenzo(a,h)anthrancene were selected to investigate as they covered the susceptible and recalcitrant PAHs in soil. A parallel experiments were also carried out on the microbial degradation of these PAHs in soil with and without biostimulation (by adding (NH4)2HPO4). The rapidly desorbed fraction of fluoranthene, as indicated by the two-fraction model, was found the highest, ranging from 21.4% to 37.4%, whereas dibenzo(a,h)anthrancene was the lowest, ranging from 8.9% to 20.5%. The rapid desorption of selected PAHs was found to be finished within 24 h. The rapidly desorbed fraction of PAHs investigated using TLPS, was significantly correlated (R2 = 0.95) with that degraded by microorganisms in biostimulation treatment. This suggested that the TLPS-assisted extraction could be a promising technique in determining the bioavailability of aged PAHs in contaminated soils. It also suggested that applying sufficient nutrients in bioremediation of field contaminated soils is crucial. Further work is required to test its application to more hydrophobic organic pollutants in long-term contaminated soils.

The Prognostic Role of Ki-67/MIB-1 in Upper Urinary-Tract Urothelial Carcinomas: A Systematic Review and Meta-Analysis.

BACKGROUND: Upper urinary-tract urothelial carcinomas (UTUC) constitute 5% of urothelial malignancies. Prognostic biomarkers would allow lower risk surgical approaches for less aggressive UTUCs. One biomarker-Ki-67/mindbomb E3 ubiquitin protein ligase 1 (Ki-67/MIB-1)-shows promise in UTUC, but there have been conflicting findings regarding its prognostic role. The systematic review and meta-analysis aim to determine the prognostic value of Ki-67/MIB-1 in UTUC in terms of UTUC-specific mortality rate, 5-year disease-free survival, and 5-year overall survival (including disease-specific survival). METHODS: A systematic review of the current literature produced 654 records. A total of 13 studies consisting of 1030 patients were finally included in the meta-analysis. Hazard ratios (HRs) with 95% confidence intervals (CI) were extracted or estimated. The individual HR estimates were combined into a pooled HR using a fixed-effects model that summed homogeneity of the individual true HRs. RESULTS: Patients with Ki-67/MIB-1 overexpression displayed significantly higher UTUC-specific mortality rate (pooled HR: 2.14, 95% CI: 1.73-2.64; p<0.00001), significantly reduced 5-year disease-free survival (pooled HR: 2.27, 95% CI: 1.79-2.92; p<0.00001), and significantly reduced 5-year overall survival (pooled HR=1.77; 95% CI: 1.39-2.23 p<0.00001). There was significant heterogeneity detected in the UTUC-specific mortality rate meta-analysis (I(2)=63%) and the 5-year disease-free survival meta-analysis (I(2)=65%), but there was no significant heterogeneity detected in the 5-year overall survival meta-analysis (I(2)=0%). Egger's testing showed that none of the outcomes were influenced by publication bias (p>0.05). CONCLUSIONS: Ki-67/MIB-1 overexpression shows promise as a prognostic biomarker for UTUC patients and requires further investigation.

Ratiometric emission fluorescent pH probe for imaging of living cells in extreme acidity.

A novel ratiometric emission fluorescent probe, 1,1-dimethyl-2-[2-(quinolin-4-yl)vinyl]-1H-benzo[e]indole (QVBI), is facilely synthesized via ethylene bridging of benzoindole and quinoline. The probe exhibits ratiometric fluorescence emission (F(522nm)/F(630nm)) characteristics with pKa 3.27 and linear response to extreme-acidity range of 3.8-2.0. Also, its high fluorescence quantum yield (Φ = 0.89) and large Stokes shift (110 nm) are favorable. Moreover, QVBI possesses highly selective response to H(+) over metal ions and some bioactive molecules, good photostability, and excellent reversibility. The probe has excellent cell membrane permeability and is further applied successfully to monitor pH fluctuations in live cells and imaging extreme acidity in Escherichia coli cells without influence of autofluorescence and native cellular species in biological systems.

miR-101 suppresses vascular endothelial growth factor C that inhibits migration and invasion and enhances cisplatin chemosensitivity of bladder cancer cells.

BACKGROUND: The microRNA miR-101 is downregulated in several cancers, including bladder cancer. However, miR-101's role in the invasion, metastasis, and chemosensitivity of bladder cancer cells remains unclear. This study was conducted to determine miR-101's role on the lymphangiogenic molecule vascular endothelial growth factor C (VEGF-C) and their effects upon bladder cancer cell migration, invasion, and chemosensitivity to cisplatin. METHODS: Two bladder cancer cell lines (T24 and 5637) and the tool cell line 293T were employed here. Bladder cancer cells were transfected with either a miR-101 overexpression vector or a scrambled-sequence lentivirus, both of which exhibited a high transfection efficiency. Non-transfection was used as a mock negative control. Wound healing and Transwell assays were performed to measure cell migration and invasiveness. A luciferase reporter assay was performed to validate miR-101's interaction with VEGF-C's 3' untranslated region followed by RT-PCR and Western blot confirmation. An MTS assay was used to evaluate the cisplatin sensitivity of the cell lines. RESULTS: miR-101 overexpression significantly inhibited the migration and invasiveness while significantly enhancing cisplatin sensitivity. miR-101 negatively regulated VEGF-C protein expression, and VEGF-C overexpression rescued the effects of miR-101 overexpression, indicating that miR-101 negatively regulates VEGF-C protein expression post-transcriptionally. miR-101 and VEGF-C interference independently enhanced cisplatin cytotoxicity in bladder cancer cells. CONCLUSIONS: miR-101 suppresses VEGF-C expression, inhibits cell migration and invasion, and increases cisplatin sensitivity in bladder cancer cells. This study provides new insight into miR-101's role in bladder cancer and shows miR-101's promise as a potential molecular target for bladder cancer.

Low temperature synthesis of phosphorous and nitrogen co-doped yellow fluorescent carbon dots for sensing and bioimaging.

Phosphorous and nitrogen co-doped carbon dots (P,N-CDs) with satisfactory quantum yield have been prepared through one-step acidic oxidation of pumpkin by H3PO4 at low temperature (90 °C). The as-prepared P,N-CD is relatively monodisperse with a narrow size distribution. The P,N-CD displays a remarkable emission enhancement in the yellow fluorescence region (λem = 550 nm) when the pH is increased from 1.5 to 7.4. The pKa value of P,N-CDs was found to be 4.17 and it shows linear response to the physiological range of pH 4.7-7.4, which is valuable for near-neutral cytosolic pH research. It is observed that P,N-CDs are superior fluorescent bioimaging agents in animals and cells thanks to their excellent solubility and ultra-low toxicity. In addition, P,N-CDs display a notably large Stokes shift of 125 nm, good reversibility and could effectively avoid the influence of autofluorescence in biological systems. The confocal fluorescent microscopic images of subcellular distribution and the detection of pH in MCF-7 cells were achieved successfully, suggesting that P,N-CDs have excellent cell membrane permeability and are further applied successfully to monitor pH fluctuations in live cells with negligible autofluorescence.