Upregulation of miR-142-5p induced by lipopolysaccharide contributes to apoptosis of hepatocytes and hepatic failure.

OBJECTIVE: This study was probed to uncover the mechanism of miR-142-5p in septic liver injury. MATERIALS AND METHODS: In this study, in-vitro and in-vivo models of sepsis were used. For in-vitro sepsis model, hepatocyte cell line (L02 cells) was treated with LPS (lipopolysaccharide). Whereas for in-vivo sepsis model, cecal ligation and puncture were performed in mice. Mice were assigned into three groups: control, CLP (Cecal Ligation Puncture), CLP + miR-142-5p inhibitor group. Liver injury was assessed via H&E staining. IL-6, TNF-α, and IL-1ß expressions were assayed through ELISA kits. C-caspase-9, C-caspase-3, ERK, p65, and IκBα expressions were determined via western blot and RT-qPCR. Apoptosis in LPS-induced L02 cells was detected by TUNEL staining. RESULTS: Our results show that miR-142-5p exhibited perspicuous upregulation in CLP mice tissues and LPS-induced L02 cells. On the other hand, inhibition of miR-142-5p could promote LPS-induced L02 cell activity and reduce apoptosis and inflammation. In terms of molecular mechanism, downregulation of miR-142-5p could abate sepsis-mediated acute hepatic injury by targeting SOCS1, through ERK and NF-κB pathway. CONCLUSIONS: Overall our results demonstrate that miR-142-5p inhibitors can mitigate septic liver injury by downregulating the inflammation and apoptosis via targeting SOCS1. Thus, miR-142-5p can serve a potential therapeutic target for sepsis mediated acute hepatic injury.

Prognostic potentials of miRNA-19a-3p and PDCD5 in nasopharynx carcinoma.

OBJECTIVE: To determine expressions of MicroRNA-19a-3p (miRNA-19a-3p) and PDCD5 in nasopharyngeal carcinoma (NPC) tissues, and their prognostic potentials in NPC. PATIENTS AND METHODS: Expressions of miRNA-19a-3p and PDCD5 in NPC tissues and controls were determined by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). The correlation between expressions of miRNA-19a-3p and PDCD5 in NPC was evaluated by Pearson correlation test. Furthermore, potential influences of miRNA-19a-3p and PDCD5 on clinical features of NPC patients were assessed. Through 5-year follow-up, survival analysis in NPC patients was conducted by Kaplan-Meier method. Finally, factors influencing prognosis of NPC were determined using the Cox regression model. RESULTS: MiRNA-19a-3p was upregulated and PDCD5 was downregulated in NPC tissues. Pearson correlation test uncovered a negative correlation between expression levels of miRNA-19a-3p and PDCD5 in NPC tissues. MiRNA-19a-3p level was correlated with N classification and clinical stage in NPC patients, while PDCD5 level was correlated with T classification, pathological grade and clinical stage. Survival analysis showed poor prognosis in NPC patients expressing high level of miRNA-19a-3p or low level of PDCD5. Cox regression analysis illustrated that N2+3 classification, clinical stage III+IV, high level of miRNA-19a-3p and low level of PDCD5 were independent risk factors for the prognosis of NPC. CONCLUSIONS: MiRNA-19a-3p is upregulated and PDCD5 is downregulated in NPC tissues. High level of miRNA-19a-3p and low level of PDCD5 are unfavorable for the prognosis of NPC.

LncRNA CCAT1 promotes cell proliferation and differentiation via negative modulation of miRNA-218 in human DPSCs.

OBJECTIVE: To investigate the expression of Long non-coding RNA (LncRNA) CCAT1 and potential functions in promoting cell proliferation and differentiation, via miRNA-218 in human adult Dental Pulp Stem Cells (DPSCs). PATIENTS AND METHODS: CCAT1 expressions in Periodontal Ligament Cells (PDLCs), DPSCs, differentiated main population (MP) cells and stem-cell-enriched Side Population (SP) cells in DPSCs were detected by qRT-PCR. MTT assay and ELISA assay were performed to evaluate the DPSCs cell proliferation and differentiation. The correlation between miR-218 and CCAT1 was detected by statistical analysis. The bioinformatics and luciferase assay were performed to explore the interaction and binding site of CCAT1 and miR-218. RESULTS: Results showed the CCAT1 expression was up-regulated in DPSCs cells. And the expression level in MP cell was higher than SP cell. MTT assay and showed overexpression CCAT1 significantly increased cell proliferation of DPSCs. ELISA assay showed the expressions of collagen I, Osteopontin (OPN) and Osteocalcin (OCN) were significantly increased in DPSCs compared with control (p<0.05). The bioinformatics and luciferase assay showed that the CCAT1 directly interacted with miR-218. In addition, miR-218 expression was negatively correlated with CCAT1 expression in DPSCs CONCLUSIONS: For the first time, we found that lncRNA-CCAT1 was upregulated in DPSCs, which could promote cell proliferation and differentiation by repressing the expression of miR-218.

NLRP12 promotes host resistance against Pseudomonas aeruginosa keratitis inflammatory responses through the negative regulation of NF-κB signaling.

OBJECTIVE: To investigate the role of NLRP12 in regulating Pseudomonas aeruginosa (P. aeruginosa) keratitis. MATERIALS AND METHODS: Real-Time-PCR and Western blot were performed to measure the NLRP12 level in corneas and bone marrow-derived macrophages (BMDMs) of C57BL/6 (B6) mice. B6 mice received a subconjunctival injection of lentivirus expressing active NLRP12 (NLRP12-lentivirus) or Ctl-lentivirus (as control), followed by infection of P. aeruginosa. The clinical score, slit lamp and bacterial plate count of mice were evaluated. In addition, myeloperoxidase (MPO) was detected to assess the infiltration of polymorphonuclear neutrophil (PMN). Cytokine levels were measured by Real Time-PCR and ELISA. Meanwhile, the bacterial burden was also evaluated. The activation of NF-κB signaling was determined by pIκBα/IκBα levels based on Western blot and NF-κB-dependent Luciferase activity on the basis of Luciferase assays using 293T cells. RESULTS: NLRP12 mRNA and protein levels were decreased in B6 corneas and BMDMs after P. aeruginosa infection. The over-expression of NLRP12 in B6 corneas significantly ameliorated the severity of corneal disease, bacterial burden, PMN infiltration and pro-inflammatory cytokine expression. In vitro analysis demonstrated that the up-regulation of NLRP12 suppressed pro-inflammatory cytokine production and enhanced bacterial clearance in RAW264.7 cells. The protein levels of pIκBα and IκBα were significantly decreased after NLRP12-lentivirus treatment compared with that of Ctl-lentivirus. NF-κB-dependent Luciferase activity was potently inhibited by NLRP12 infected with P. aeruginosa or cotransfected with the downstream signaling molecules including IKKα and IKKß in 293T cells. CONCLUSIONS: NLRP12 decreases the severity of P. aeruginosa keratitis, reduces corneal inflammation and bacterial burden through the down-regulation of the NF-κB signaling pathway.

The expressions and significance of APN, D-D, IL-17 and hs-CRP in patients with acute exacerbation of chronic obstructive pulmonary disease.

OBJECTIVE: Inflammatory reactions and imbalance of oxidant/antioxidant and protease/anti-protease are the major causes of chronic obstructive pulmonary disease. Based on the information mentioned, the expressions and significance of adiponectin (APN), D-dimer (DD), Interleukin (IL)-17, and high-sensitivity CRP (hs-CRP) in patients with acute exacerbation of chronic obstructive pulmonary disease were investigated in this study. PATIENTS AND METHODS: A total of 70 patients with chronic obstructive pulmonary disease were enrolled and divided into stable group (group A, 28 cases) and acute exacerbation group (group B, 42 cases). Thirty-five healthy volunteers were included in the control group (group C, 35 cases). The levels of serum APN, IL- 17, D-D, and hs-CRP were tested and compared RESULTS: Levels of APN from Group B were significantly lower than that of Group A or Group C, while levels of APN of Group A were also significantly lower than that of Group C, (p < 0.05). Levels of IL-17, D-D, and Hs-CRP of group b were significantly increased compared to that of Group A or Group C, and levels of IL-17, D-D, and Hs-CRP of Group A were significantly elevated compared to that of Group C (p < 0.05). A negative statistical correlation was found between APN and IL-17, D-D, and Hs-CRP (p < 0.05). CONCLUSIONS: Levels of APN were downregulated in patients with acute exacerbation of chronic obstructive pulmonary disease. The expression levels of APN, IL-17, D-D, and Hs-CRP were closely correlated with clinical stages and can be used as parameters for the evaluation of the severity of chronic obstructive pulmonary disease.

MiRNA regulates OCT4 expression in breast cancer cells.

OBJECTIVE: Breast cancer cell infiltration, migration, and proliferation significantly affect its curative effect. Stemness gene octamer-binding transcription factor 4 (OCT4) upregulated in breast cancer tissue compared with normal control. MiRNA exhibits regulatory role in gene expression. This study adopted bioinformatics to predict the miRNA to regulate OCT4 gene and investigated its impact on breast cancer cell infiltration, migration, and proliferation. MATERIALS AND METHODS: MirBase database was analyzed to explore the potential miRNA in regulating OCT4 based on human OCT4 gene sequence. MiRNA mimics and inhibitor were synthetized and transfected to BS524 cells. qRT-PCR was applied to test miRNA and OCT4 mRNA expressions in cells at 12 h, 24 h, and 48 h after transfection. Western blot was selected to detect OCT4 protein expression. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was selected to determine cell proliferation. Scratch assay was adopted to evaluate cell migration. Transwell assay was used to analyze cell infiltration. RESULTS: MiR-145 may regulate OCT4 gene with score 82. OCT4 mRNA and protein increased at 12 h after transfection (p > 0.05). OCR4 gene significantly upregulated, cell proliferation, migration, and infiltration enhanced by miR-145 transfection compared with control (p < 0.05). OCT4 gene downregulated, while cell proliferation, infiltration, and migration markedly weakened in miR-145 inhibitor group compared with control (p < 0.05). CONCLUSIONS: MiR-145 affects breast cancer BS524 cell proliferation, infiltration, and migration via positively regulating OCT4 gene expression.

Analysis of nasal cavity morphology and nasolabial development of the normal Han ethnic people under age of 12.

OBJECTIVE: This study aims to establish the three-dimensional (3D) reconstruction model of nasal cavity for China's Han ethnic population (0-12 years) by laser scanning and photogrammetry, and thus to elucidate the developmental mechanism of nasal cavity morphology and nasolabial region. PATIENTS AND METHODS: A total of 260 normal people of the Han ethnic aged 0-12 were recruited as subjects, among whom 60 were scanned for nasal cavity morphology in order to get reconstructed models with the computer engineering software. Photogrammetry was performed for the remaining 200 subjects to measure the 7 parameters that reflect vertically or horizontally the anatomical features of the nasolabial region. RESULTS: The interior morphology of nasal cavity was accurately established by 3D laser scanning and photogrammetry with the optimal morphology of nasal cavity simulated through 3D reconstruction. Development of nasal cavity and nasolabial region was also analyzed. CONCLUSIONS: The 3D laser scanning analysis is the ideal method to analyze the interior morphology of nasal cavity by reconstructing the normal interior morphology of nasal cavity and quantitatively analyze the change of nasal cavity morphology with age. Photogrammetry can be applied to conduct the morphological measurement for the nasolabial region and, thus, assessing the development of the nasolabial region with age, which provides information for choosing the timing and options of surgery in treating harelip and nasal deformity.

Carboxypeptidase E protects hippocampal neurons during stress in male mice by up-regulating prosurvival BCL2 protein expression.

Prolonged chronic stress causing elevated plasma glucocorticoids leads to neurodegeneration. Adaptation to stress (allostasis) through neuroprotective mechanisms can delay this process. Studies on hippocampal neurons have identified carboxypeptidase E (CPE) as a novel neuroprotective protein that acts extracellularly, independent of its enzymatic activity, although the mechanism of action is unclear. Here, we aim to determine if CPE plays a neuroprotective role in allostasis in mouse hippocampus during chronic restraint stress (CRS), and the molecular mechanisms involved. Quantitative RT-PCR/in situ hybridization and Western blots were used to assay for mRNA and protein. After mild CRS (1 h/d for 7 d), CPE protein and mRNA were significantly elevated in the hippocampal CA3 region, compared to naïve littermates. In addition, luciferase reporter assays identified a functional glucocorticoid regulatory element within the cpe promoter that mediated the up-regulation of CPE expression in primary hippocampal neurons following dexamethasone treatment, suggesting that circulating plasma glucocorticoids could evoke a similar effect on CPE in the hippocampus in vivo. Overexpression of CPE in hippocampal neurons, or CRS in mice, resulted in elevated prosurvival BCL2 protein/mRNA and p-AKT levels in the hippocampus; however, CPE(-/-) mice showed a decrease. Thus, during mild CRS, CPE expression is up-regulated, possibly contributed by glucocorticoids, to mediate neuroprotection of the hippocampus by enhancing BCL2 expression through AKT signaling, and thereby maintaining allostasis.

Type-Two intramolecular diels-alder reactions of pyrazolo-o-quinodimethanes

Two isomeric series of homologous N-(acryloyloxy)alkylated pyrazolo-3-sulfolenes 10a-c and 18a-c have been efficiently synthesized from a common starting material, 4,6-dihydro-1H-thieno[3, 4-c]pyrazole (5). Thermolysis of these fused 3-sulfolenes provides the corresponding o-quinodimethanes which simultaneously undergo "type-two" intramolecular Diels-Alder reactions to form two- and three-atom-bridged tricyclic pyrazoles which are otherwise difficult to prepare. It was also demostrated that, depending on the N-substitution position of the pyrazolo-fused 3-sulfolenes, the temperature required for the thermal extrusion of SO(2) and the regioselectivity of the T2-IMDA reactions were influenced substantially.

Long-range through-bond photoactivated sigma bond cleavage in steroids. Intramolecular sensitized debromination

[structure: see text] The photolysis of 17alpha-bromo-3alpha-(triphenylsilyloxy)-5alpha-androstane (2; 3alphaTPSO/17alphaBr) and 17alpha-bromo-3alpha-(triphenysilyloxy)-5-androstan-6-one (3; 3alphaTPSO/6ketone/17alphaBr) is described. Irradiation of 2 with 266 nm light leads to debromination via intramolecular transfer of triplet excitation energy with a quantum efficiency of 0.0011. Photolysis of 3 with both 266 and 308 nm light leads to debromination with quantum efficiencies of ca. 0.0066. The debromination of 3 is attributed to activation via the ketone excited singlet state, with singlet energy transfer from C6 to C17 ca. 35% efficient and occurring with a rate constant of 1.4 x 10(8) s(-1).