Scopolamine causes delirium-like brain network dysfunction and reversible cognitive impairment without neuronal loss.

Delirium is a severe acute neuropsychiatric syndrome that commonly occurs in the elderly and is considered an independent risk factor for later dementia. However, given its inherent complexity, few animal models of delirium have been established and the mechanism underlying the onset of delirium remains elusive. Here, we conducted a comparison of three mouse models of delirium induced by clinically relevant risk factors, including anesthesia with surgery (AS), systemic inflammation, and neurotransmission modulation. We found that both bacterial lipopolysaccharide (LPS) and cholinergic receptor antagonist scopolamine (Scop) induction reduced neuronal activities in the delirium-related brain network, with the latter presenting a similar pattern of reduction as found in delirium patients. Consistently, Scop injection resulted in reversible cognitive impairment with hyperactive behavior. No loss of cholinergic neurons was found with treatment, but hippocampal synaptic functions were affected. These findings provide further clues regarding the mechanism underlying delirium onset and demonstrate the successful application of the Scop injection model in mimicking delirium-like phenotypes in mice.

How well do integrated 3D models predict alveolar defects after treatment with clear aligners?

OBJECTIVES: To evaluate the accuracy of integrated models (IMs) constructed by pretreatment cone-beam computed tomography (pre-CBCT) in diagnosing alveolar defects after treatment with clear aligners. MATERIALS AND METHODS: Pre-CBCT and posttreatment cone-beam computed tomography (CBCT) scans from 69 patients who completed nonextraction treatment with clear aligners were collected. The IMs comprised anterior teeth in predicted positions and alveolar bone from pre-CBCT scans. The accuracy of the IMs for identifying dehiscences or fenestrations was evaluated by comparing the means of the defect volumes, absolute mean differences, and Pearson correlation coefficients with those measured from post-CBCT scans. Defect prediction accuracy was assessed by sensitivity, specificity, positive predictive values, and negative predictive values. Factors possibly affecting changes in mandibular alveolar defects were analyzed using a mixed linear model. RESULTS: The IM measurements showed mean deviations of 2.82 ± 9.99 mm3 for fenestrations and 3.67 ± 9.93 mm3 for dehiscences. The absolute mean differences were 4.50 ± 9.35 mm3 for fenestrations and 5.17 ± 9.24 mm3 for dehiscences. The specificities of the IMs were higher than 0.8, whereas the sensitivities were both lower (fenestration = 0.41; dehiscence = 0.53). The positive predictive values were unacceptable (fenestration = 0.52; dehiscence = 0.62), and the overall reliability was low (<0.80). Molar distalization and proclination were positively correlated with significant increases in alveolar defects at the mandibular incisors after treatment. CONCLUSIONS: Alveolar defects after clear aligner treatment cannot be simulated accurately by IMs constructed from pre-CBCT. Caution should be taken in the treatment of crowding with proclination and molar distalization for the safety of alveolar bone at the mandibular incisors.

[Characteristic expression of apoptotic genes in epulis].

PURPOSE: To investigate the characteristic expression of apoptotic genes in epulis. METHODS: Thirty-six patients with epulis were recruited in this study, and related tumor and normal gingival tissues were collected. Six pairs of samples were used for PCR array analysis of apoptosis, and the other thirty pairs of samples were used for qRT-PCR replication. The expression levels of related genes were calculated with internal reference gene delta ΔΔCt. The data were analyzed with SPSS 17.0 software package. RESULTS: Compared with the normal gingival tissues, the expression levels of AIFM1, BCL2, BCL2L1, BCL2L2, BFAR, BIRC2, BIRC3, BIRC6, BNIP2, BNIP3, CD40LG, and XIAP were significantly increased in the gingival tissues, while the expression level of TNFRSF25 was decreased. CONCLUSIONS: Over-expression of anti-apoptotic genes in Bcl-2 family and IAP family inhibits apoptosis of gingival tissues, which eventually causes epulis.

Lentiviral­mediated Shh reverses the adverse effects of high glucose on osteoblast function and promotes bone formation via Sonic hedgehog signaling.

Patients with diabetes tend to have an increased incidence of osteoporosis, which may be associated with hyperglycemia; however, the pathogenic mechanisms governing this interaction remain unknown. The present study sought to investigate whether elevated extracellular glucose levels of bone mesenchymal stem cells (BMSCs) could influence osteoblastic differentiation and whether the intracellular Sonic hedgehog (Shh) pathway could adjust the effects. Furthermore, to verify the results in vivo, a rat tooth extraction model was constructed. BMSCs were incubated in eight types of culture medium, including low glucose (LG), LG + lentivirus (Lenti), LG + Lenti­small interfering RNA (Lenti­siRNA), LG + Lenti­Shh, high glucose (HG), HG + Lenti, HG + Lenti­siRNA and HG + Lenti­Shh. The lentiviral transfection efficiency was observed using a fluorescence microscope; protein and mRNA expression was detected by western blotting and reverse transcription­quantitative polymerase chain reaction (RT­qPCR). The matrix mineralization and alkaline phosphatase (ALP) activity of BMSCs were examined by Alizarin red staining and ALP activity assays, respectively. The expression of osteogenesis­related genes in BMSCs were quantified by RT­qPCR. The alveolar ridge reduction was measured and histological sections were used to evaluate new bone formation in the tooth socket. With high concentrations of glucose, Shh expression, matrix mineralization nodules formation, ALP activity and the levels of bone morphogenic protein 4 (BMP4), bone sialoprotein (BSP) and osteopontin (OPN) expression were greatly reduced compared with LG and corresponding control groups. Whereas activated Shh signaling via Lenti­Shh could increase the number of matrix mineralization nodules, ALP activity, and the expression levels of BMP4, BSP and OPN in BMSCs. Additionally, in vivo assays demonstrated that Lenti­Shh induced additional bone formation. Collectively, the results of the present study indicated that HG inhibited the Shh pathway in osteoblasts and resulted in patterning defects during osteoblastic differentiation and bone formation, while the activation of Shh signaling could suppress these deleterious effects.

[Analysis of orthologous lncRNAs in humans and mice and their species-specific epigenetic target genes].

OBJECTIVE: To identify orthologous lncRNAs in human and mice and the species specificity of their epigenetic regulatory functions. METHODS: The human/mouse whole-genome pairwise alignment (hg19/mm10, genome.UCSC.edu) was used to identify the orthologues in 13 562 and 10 481 GENCODE-annotated human and mouse lncRNAs. The Infernal program was used to search the orthologous sequences of all the exons of the 13562 human lncRNAs in mouse genome (mm10) to identify the highly conserved orthologues in mice. LongTarget program was used to predict the DNA binding sites of the orthologous lncRNAs in their local genomic regions. Gene Ontology analysis was carried out to examine the functions of genes. RESULTS: Only 158 orthologous lncRNAs were identified in humans and mice, and many of these orthologues had species-specific DNA binding sites and epigenetic target genes. Some of the epigenetic target genes executed important functions in determining human and mouse phenotypes. CONCLUSION: s Only a few human and mouse lncRNAs are orthologues, and most of lncRNAs are species-specific. The orthologous lncRNAs have species-specific epigenetic target genes, and species-specific epigenetic regulation greatly contributes to the differences between humans and mice.

Tension-loaded bone marrow stromal cells potentiate the paracrine osteogenic signaling of co-cultured vascular endothelial cells.

Co-culture of bone marrow stromal cells (BMSCs) and vascular endothelial cells (VECs) is a promising strategy for better osteogenesis and pre-vascularization in bone tissue engineering. Recent reports have shown that mechanical stretching further promotes osteogenesis in BMSC/VEC co-culture systems, but the underlying mechanism of this process remains unclear. In this study, noncontact co-cultures of rat primary BMSCs and VECs were employed to interrogate paracrine cell-to-cell communications in response to tension. Exposure of VECs to 6% tension for 48 h elicited neither ALP activity nor mRNA expression of OCN and OPN in BMSCs incubated in a shared culture medium. Instead, BMSCs subjected to tension induced robust VEGF release, and its conditioned medium enhanced the proliferation and tubular formation of VECs with a concurrent increase in BMP-2 and IGF-1 production. Conditioned medium from activated VECs in turn promoted expression of osteogenic genes in BMSCs, followed by an increase in matrix mineralization. The addition of VEGF-R inhibitor Tivozanib to these systems abrogated the tension-induced paracrine effects on VECs and subsequently impaired BMSC osteogenesis. These results clearly demonstrate that the response of BMSCs to tension potentiates paracrine osteogenic signaling from VECs; this positive feedback loop is initiated by VEGF release.

A novel functional polymorphism of GFAP decrease glioblastoma susceptibility through inhibiting the binding of miR-139.

Glioblastoma (GBM) is the most commonly diagnosed solid tumor outside the central nervous system. However, genetic factors underlying GBM remain largely unclear. Previous studies indicated that Glial fibrillary acidic protein (GFAP) might play an important role in the aggressiveness of GBM and also contributed to its poor overall survival. The present study aims to test (1) the associations between GFAP single nucleotide polymorphisms (SNPs) and GBM cells chemoresistance and metastasis, and (2) the molecular mechanism accounting for their effects. Four tagging SNPs of GFAP were initially genotyped in 667 subjects and the significant SNP was further analyzed via online bioinformatical tools. SNP rs11558961 was found to be significantly associated with GBM susceptibility. It was predicted to influence microRNA(miR)-139 binding to 3'UTR of GFAP gene. In functional experiments, we found that cells transfected with rs11558961 G-allele constructs had lower baseline luciferase activities and were more responsive to miR-139 changes, compared to C-allele constructs. Moreover, rs11558961 C>G variant reduced the chemoresistance of GBM cells and migration capability. In conclusion, rs11558961 might influence the chemoresistance and progression of GBM cells via promoting the binding of miR-139, ultimately decrease the susceptibility of GBM. This investigation will shed light on the optimizing for clinical trial design and individualizing of therapeutic plans.

Enhanced performance of polyimide hybrid membranes for benzene separation by incorporating three-dimensional silver-graphene oxide.

Graphene oxide-Ag nanoparticle composites were prepared through impregnation reduction using different reactants. Transmission electron microscopy, X-ray diffraction, X-ray photoelectron spectroscopy, and Fourier transform infrared spectroscopy analyses were performed to characterize differences in the morphologies of three different Ag-GO composites. Scanning electron microscopy, transmission electron microscopy, and differential scanning calorimetry analyses were also applied to evaluate the morphology and thermal stability of the hybrid membranes. Swelling-sorption and pervaporation experiments of benzene and cyclohexane were conducted to evaluate the separation performance of hybrid membranes containing different Ag-GO composites. Results demonstrated that small Ag nanoparticles generated through impregnation reduction using Ag(NH3)2(+) and PEG were homogeneously distributed in the hybrid membranes because of moderate reduction rate. The polymide (PI) hybrid membrane exhibited high separation performance. Increase in Ag content in the Ag-GO samples led to the formation of Ag particles on the GO surface; these particles enhanced the separation performance of the hybrid membranes. When Ag-GO samples with 15 mass percent added, the hybrid membrane showed the highest separation performance and its maximum separation factor in the pervaporation experiments reached 35. It is more than three times higher than that of the GO/PI hybrid membrane. Moreover, large Ag particles were formed and aggregated during the preparation and polymerization of Ag-GO samples with high Ag contents; these particles reduced the separation performance of the hybrid membranes.

TLR4 polymorphisms associated with developing gastric pre-cancer lesions in a Chinese Han population.

Helicobacter pylori infection is a risk factor for gastric cancer. In addition, toll-like receptor 4 (TLR4) plays a fundamental role in pathogen recognition and activation of innate immunity. This study investigated the association of TLR4 polymorphisms with a risk of intestinal metaplasia (IM) and intraepithelial neoplasia (IN) in a Chinese Han population. This study analyzed TLR4 gene polymorphisms in 333 patients (IM, 193 cases; IN, 140 cases) and 312 atypia-free controls in a Chinese Han population using a Taqman allelic discrimination assay. The TLR4 single nucleotide polymorphisms +896A/G and +1196C/T were not associated with the risk of IM or IN. However, the single-locus analysis showed that the C allele of TLR4+2856T/C had significantly reduced risk of IM and IN [adjusted odds ratio (OR)=0.42; 95%CI=0.29-0.62 and OR=0.62; 95%CI=0.41-0.93, respectively] compared with the wild-type homozygote (TT). The frequencies of TLR4+2856T/C TC and T carrier were significantly lower in patients with Sydney's slight IM and low grade IN (P<0.01 and P=0.01, respectively), while the TC genotype showed a lower risk of moderate IM compared to healthy controls (P=0.045). In addition, the data revealed that H. pylori infection, heavy alcohol consumption and high salt uptake were associated with a higher susceptibility for developing this neoplasm. TLR4 rs10759932 TC and C carriers were associated with a lower risk in developing precancerous lesions in the stomach in a Chinese Han population.

[Study on the pH-sensitive secondary structure of gamma-PGA embedded with magnetite nanoparticles].

The conformational transition of poly gamma-glutamic acid (gamma-PGA) embedded with magnetite nanoparticles under various pH conditions was investigated by Fourier transform infrared spectroscopy (FTIR). The secondary structure content was determined through the analysis of amide I bands of Fourier deconvolution spectra, secondary derivative spectra and the Gaussian curve fitting of the original infrared spectra. The results showed that the conformation of the gamma-PGA was affected by solution pH. The total contents of beta-sheet and beta-turn were higher than 65%, while alpha-helix and random coil were low. The content of beta-turn increased with increasing pH, while the beta-sheet decreased. Additionally, the zeta potential results showed that the pH-sensitive secondary structure of gamma-PGA had influence on the stability of suspension of magnetic gamma-PGA nanospheres. The minimum value of zeta potential (-35. 4 mV) was obtained at pH 10.2.