LINC00240 sponges miR-4465 to promote proliferation, migration, and invasion of hepatocellular carcinoma cells via HGF/c-MET signaling pathway.

OBJECTIVE: LINC00240, as a novel long non-coding RNAs (lncRNAs), has never been studied in hepatocellular carcinoma (HCC). This research reported the expression and function of LINC00240 in HCC. PATIENTS AND METHODS: LINC00240 expression in 180 HCC patients was downloaded from the Cancer Genome Atlas (TCGA) database. HCC patients' survival was analyzed via Kaplan­Meier analysis. The expression of LINC00240, miR-4465 and HGF in Hep3B and Huh7 cells were regulated by transfection. Cell viability was determined by MTT assay. Transwell experiment was used for the detection of cells migration and invasion abilities. The interaction between LINC00240, miR-4465 and HGF was reflected by Luciferase reporter assay. LINC00240, miR-4465, HGF and p-c-MET expression in HCC cells were researched by real-time quantitative polymerase chain reaction (RT-qPCR) and Western blot. RESULTS: TCGA data showed that high LINC00240 expression was markedly associated with lower survival of HCC patients (p = 0.036). LINC00240 expression was aberrantly upregulated in HCC cells. Silencing of LINC00240 significantly reduced HCC cells viability, migration and invasion. miR-4465 was a target gene of LINC00240. Silencing of LINC00240 reduced HCC cells viability, migration and invasion via directly promoting miR-4465 expression. HGF was target gene of miR-4465. miR-4465 up-regulation obviously suppressed HGF and p-c-MET expression. According to rescue experiment, LINC00240 silencing inhibited HCC cells viability, migration and invasion by suppressing HGF/c-MET signaling pathway via targeting miR-4465. CONCLUSIONS: LINC00240 sponges miR-4465 to promote HCC cells proliferation, migration and invasion via HGF/c-MET signaling pathway.

MiR-144 inhibits colorectal cancer cell migration and invasion by regulating PBX3.

OBJECTIVE: MicroRNAs (miRNA) are aberrantly expressed in various human cancers, including colorectal cancer (CRC). We aim to investigate the functional role and underlying mechanism of miR-144 in CRC. PATIENTS AND METHODS: The expressional level of miR-144 and pre-leukemia transcription factor 3 (PBX3) in CRC tissues and cells was confirmed by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) and Western blot. The migration and invasion of CRC cells were detected by transwell assay. Luciferase reporter assay was performed to determine the specific target of miR-144 in CRC cells. RESULTS: The results displayed that miR-144 expression was significantly decreased in CRC tissues and cells compared to that in normal controls. Additionally, miR-144 mimic suppressed, while miR-144 inhibitor promoted the ability of CRC cell migration and invasion. More importantly, PBX3 was the direct target of miR-144 in regulating CRC development and PBX3 could reverse the inhibitory effect of miR-144 mimic on CRC cells. PBX3 expression was significantly increased in CRC and negatively correlated with miR-144 expression. CONCLUSIONS: In conclusion, miR-144 suppressed CRC cell migration and invasion by targeting PBX3, suggesting its potential value in the diagnosis and treatment of CRC.

A modified longitudinal incision via transoral sublabial approach for removal of the nasal vestibular cyst: a clinical observation.

OBJECTIVE: The aim of this study was to investigate the clinical effect of the removal of nasal vestibular cysts through a modified longitudinal incision via a transoral sublabial approach. METHOD: In 28 cases, a nasal vestibular cyst was removed through a modified longitudinal incision via a transoral sublabial approach. A visual analogue scale score was used to evaluate the numbness of the nasal alar and upper lip. Post-operative complications were recorded. Medical photographs were used for assessment. RESULTS: For all patients, incisions reached clinical primary healing one week after surgery. All patients were free of post-operative haematoma, infection, oronasal fistula and malformation. In the first week and the first month after surgery, numbness of the nasal alar and upper lip was recorded in few cases. The patients were followed up for 2-57 months without recurrence. CONCLUSION: Removal of nasal vestibular cysts via a transoral sublabial approach with a modified longitudinal incision is a minimally invasive and simple surgical method with few complications and a quick recovery.

Research on cardioprotective effect of irbesartan in rats with myocardial ischemia-reperfusion injury through MAPK-ERK signaling pathway.

OBJECTIVE: The aim of this study was to explore whether the mechanism of Irbesartan (IRB) in the treatment of rats with myocardial ischemia-reperfusion injury (MIRI) was related to the mitogen-activated protein kinase (MAPK)-extracellular signal-regulated kinase (ERK) signaling pathway. MATERIALS AND METHODS: The rat model of MIRI was first successfully established. All rats were randomly divided into 5 groups, including the Sham group (sham operation control group, ligation only), Model group (MIRI rat model group), IRB12.5 group [low-dose IRB (12.5 mg/kg/d) group], IRB50.0 group [medium-dose IRB (50.0 mg/kg/d) group] and IRB200.0 group [high-dose IRB (200.0 mg/kg/d) group]. After treatment of IRB in MIRI rats, the activities of four myocardial enzyme indexes, including creatine kinase-MB (CK-MB), aspartate aminotransferase (AST), lactate dehydrogenase (LDH) and cardiac troponin T (cTnT), were detected via enzyme-linked immunosorbent assay (ELISA). The effect of IRB on myocardial apoptosis in MIRI rats was detected via Annexin V-fluorescein isothiocyanate/Propidium Iodide (FITC/PI) double staining. Meanwhile, the messenger ribonucleic acid (mRNA) levels of ERK, B-cell lymphoma-2 (Bcl-2) and Bcl-2 associated X protein (Bax) in myocardial cells after treatment of IRB were detected via Reverse Transcription-Polymerase Chain Reaction (RT-PCR). Furthermore, the protein levels of ERK and p-ERK were detected via Western blotting. RESULTS: Different concentrations of IRB could protect myocardium from MIRI. IRB at doses of 50.0 mg/kg/d and 200.0 mg/kg/d could significantly downregulate myocardial enzyme indexes in MIRI (p<0.01). Meanwhile, both the doses could markedly inhibit myocardial apoptosis in MIRI rat model by regulating the expressions of apoptosis-related genes (Bcl-2 and Bax) (p<0.01), eventually improving myocardial pathological damage. At the same time, it could also significantly decrease the mRNA and protein levels of ERK in the MAPK-ERK signaling pathway (p<0.05). CONCLUSIONS: The cardioprotective mechanism of IRB in MIRI rats may be related to the inhibition of the activation of the MAPK-ERK signaling pathway.

Circular RNA in blood corpuscles combined with plasma protein factor for early prediction of pre-eclampsia.

OBJECTIVE: To determine the expression of circular RNA (circRNA) in blood corpuscles of pregnant women before 20 weeks of pregnancy and to create a new model to identify the performance of circRNA combined with protein factors for the early diagnosis of pre-eclampsia (PE). DESIGN: Nested case-control study. SETTING: University medical centre, Guangzhou, China. POPULATION: A total of 1400 pregnant women recruited between 8 and 20 weeks of gestation. In all, 41 women with PE were included in the study, and were matched with 41 normally pregnant women based on maternal age and gestational age at same-size ratio. METHODS: The samples were analysed using a human circRNA microarray in the discovery phase, then the circRNA and the plasma protein factor endoglin (ENG) were validated. Finally we combined ENG with circRNA to create a new early prediction model for PE. MAIN OUTCOME MEASURES: Early changes of circRNA and ENG in PE. RESULTS: The circ_101222 levels in blood corpuscles of patients with PE were significantly higher than those in corresponding healthy women (P < 0.001). Using ENG in combination with circ_101222 resulted in a sensitivity of 0.7073, a specificity of 0.8049, and overall area under the curve of 0.876 (95% confidence interval 0.816-0.922) for the prediction of PE. CONCLUSION: CircRNA and plasma proteins may have some predictive value for PE (such as circ_101222 and ENG). The performance of each of these factors may be strengthened when plasma proteins are used in combination with circRNA. The results are preliminary and need to be validated in larger studies and other populations. TWEETABLE ABSTRACT: Plasma protein endoglin in combination with circ_101222 strengthened the predictive power for pre-eclampsia.

The regulatory role of NF-κB in autophagy-like cell death after focal cerebral ischemia in mice.

Autophagy may contribute to ischemia-induced cell death in the brain, but the regulation of autophagic cell death is largely unknown. Nuclear factor kappa B (NF-κB) is a regulator of apoptosis in cerebral ischemia. We examined the hypothesis that autophagy-like cell death could contribute to ischemia-induced brain damage and the process was regulated by NF-κB. In adult wild-type (WT) and NF-κB p50 knockout (p50(-/-)) mice, focal ischemia in the barrel cortex was induced by ligation of distal branches of the middle cerebral artery. Twelve to 24h later, autophagic activity increased as indicated by enhanced expression of Beclin-1 and LC3 in the ischemic core and/or penumbra regions. This increased autophagy contributed to cell injury, evidenced by terminal deoxynucleotidyltransferase (TdT)-mediated dUTP-biotin nick end labeling (TUNEL) co-staining and a protective effect achieved by the autophagy inhibitor 3-methyladenine. The number of Beclin-1/TUNEL-positive cells was significantly more in p50(-/-) mice than in WT mice. Neuronal and vascular cell death, as determined by TUNEL-positive cells co-staining with NeuN or Collagen IV, was more abundant in p50(-/-) mice. Immunostaining of the endothelial cell tight junction marker occludin revealed more damage to the blood-brain barrier in p50(-/-) mice. Western blotting of the peri-infarct tissue showed a reduction of Akt-the mammalian target of rapamycin (mTOR) signaling in p50(-/-) mice after ischemia. These findings provide the first evidence that cerebral ischemia induced autophagy-like injury is regulated by the NF-κB pathway, which may suggest potential treatments for ischemic stroke.

Percutaneous anterior transarticular screw fixation for atlantoaxial instability: a case series.

We reviewed the outcome of a retrospective case series of eight patients with atlantoaxial instability who had been treated by percutaneous anterior transarticular screw fixation and grafting under image-intensifier guidance between December 2005 and June 2008. The mean follow-up was 19 months (8 to 27). All eight patients had a solid C1-2 fusion. There were no breakages or displacement of screws. All the patients with pre-operative neck pain had immediate relief from their symptoms or considerable improvement. There were no major complications. Our preliminary clinical results suggest that percutaneous anterior transarticulation screw fixation is technically feasible, safe, useful and minimally invasive when using the appropriate instruments allied to intra-operative image intensification, and by selecting the correct puncture point, angle and depth of insertion.

Offspring from heterotopic transplantation of newborn mice ovaries.

This study is aimed at investigating the developmental potential of the primordial follicles from ovaries of newborn mice after cryopreservation in liquid nitrogen for long-term storage, thawing, and heterografting into the kidney capsules of ovariectomized adult female mice. After stimulation of recipient mice with pregnant mare serum gonadotropin on day-19 after heterografting, the primordial follicles of the transplanted ovaries could develop into antral follicles. When the oocyte-cumulus cell complexes were retrieved from these antral follicles, they could mature after in vitro culture for 16­17 h. After in vitro fertilization, the rates of embryos derived from these oocytes that developed into the two-cell stage and the blastocyst stage after 16­17 h and after day-4, respectively,in the culture medium were 55.40% (55/107) and 9.09% (5/55),respectively. In the ovarian transplantation groups, no pups were derived from the 410 embryos that were transferred into 10 pseudopregnant mothers at the pronuclear stage. However,of the 10 surrogate mothers in whom 570 embryos were transferred at the two-cell stage, four achieved pregnancy and gave birth to 20 live offspring. These results demonstrated that primordial follicles in newborn mice ovaries were capable of sustaining their developmental potential after freezing and thawing. Once transplanted into the kidney capsules of ovariectomized adult female mice, these primordial follicles could develop and respond to gonadotropin stimulation and reach the antral stage; further, live offspring could be derived from these follicles.

The structures of mouse and human L1 elements reflect their insertion mechanism.

L1 is an abundant, interspersed repeated DNA element of mammalian genomes. It has achieved its high copy number via retrotransposition. Like other non-LTR retrotransposons, L1 insertion into chromosomal DNA apparently occurs by target-site primed reverse transcription, or TPRT. L1 retrotransposition often generates elements with 5' truncations that are flanked by a duplication of the genomic target site (TSD). It is typically assumed that the 5' truncated elements are the consequence of poor processivity of the L1 reverse transcriptase. However, we find that the majority of young L1 elements from both the human and mouse genomes are truncated at sequences that can basepair with the target site. Thus, to whatever extent truncation is a consequence of poor processivity, we suggest that truncation is likely to occur when target site sequence can basepair with L1 sequence. This finding supports a model for insertion that occurs by two sequential TPRT reactions, the second of which relies upon the homology between the target site and L1. Because perfect heteroduplex formation is not required for all insertions, a dynamic relationship between the primer, template and enzyme during reverse transcription is inferred. 5' truncation may be a successful evolutionary strategy that is exploited by L1 as a means to escape host suppression of transposition.