Er:YAG laser settings for debonding zirconia restorations: An in vitro study.

This in vitro study aimed to determine the optimal frequency and energy settings for debonding zirconia restorations using an erbium-doped yttrium aluminum garnet (Er:YAG) laser. A total of 200 zirconia specimens (5 mm × 5 mm × 1.5 mm) were fabricated from two types of materials: (1) 3 mol% yttria oxide stabilized tetragonal zirconia polycrystalline (3Y-TZP) and (2) 5 mol% yttria oxide stabilized tetragonal zirconia polycrystalline (5Y-TZP). The zirconia specimens were bonded to dentin using resin cement (RelyX Ultimate, 3 M) and divided into 20 groups based on their laser treatments (n = 5). Er:YAG laser treatment was applied at various frequencies (10 Hz and 20 Hz) and energies (80 mJ, 100 mJ, 120 mJ, 140 mJ, 160 mJ, 180 mJ, 200 mJ, 220 mJ, 240 mJ, and 260 mJ). The time required to debond the specimens and the temperature changes that dentin underwent during the laser treatment were recorded. The surface morphologies of the debonded dentin and zirconia specimens were observed using scanning electron microscopy (SEM). Additional zirconia specimens were fabricated for 4-point flexural strength testing and surface roughness measurements. Statistical analyses were conducted using three-way analysis of variance (ANOVA) and Student-Newman-Keuls (SNK)-q tests (α = 0.05). The debonding time of each specimen varied between 4.8 and 160.4 s, with an average value of 59.2 s. The dentin temperature change for each specimen ranged from 2.3 to 3.6 °C, with an average value of 2.7 °C. The debonding time was significantly influenced by the zirconia material type and laser energy, but it was not affected by the laser frequency. Among the specimens, those made of 3Y-TZP needed significantly more time for debonding than 5Y-TZP. The optimal energies were 220 mJ for 3Y-TZP and 200 mJ for 5Y-TZP. The laser frequency, laser energy, and type of zirconia material had no effect on the dentin temperature change. Additionally, no surface alternations were observed on the dentin or zirconia materials after laser treatment. The surface roughness and flexural strength of the zirconia materials remained unchanged after laser treatment. In summary, Er:YAG laser treatment effectively and safely removes zirconia restorations without impacting their mechanical properties, with a safe temperature change of less than 5.6 °C. The optimum frequency and energy settings for debonding 3Y-TZP and 5Y-TZP restorations were found to be 10/20 Hz and 220 mJ and 10/20 Hz and 200 mJ, respectively.

Identifying and validating key genes mediating intracranial aneurysm rupture using weighted correlation network analysis and exploration of personalized treatment.

Background: Intracranial aneurysmal subarachnoid hemorrhage (aSAH) is a dangerous and highly fatal condition if ruptured. Significant advances have been made in the treatment of unruptured intracranial aneurysms (UIAs), but risk assessment methods for early diagnosis of intracranial aneurysm (IA) rupture remain limited. Methods: The datasets of IA GSE13353, GSE15629, and GSE54083 were downloaded through the Gene Expression Omnibus (GEO) database. Differentially expressed genes (DEGs) in unruptured and ruptured aneurysms were identified by R software using methods such as gene set enrichment analysis (GSEA) and weighted gene co-expression network analysis (WGCNA). Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis were performed on the DEGs, and logistic regression models were used to construct a prediction model to discriminate UIA from healthy samples. We then performed GSEA on the genes in the model, followed by model validation using the GSE54083 dataset. Finally, we used the single-sample (ss)GSEA method to investigate the relationship between the diagnostic model genes and immune cells and immune function. Results: A total of 79 DEGs were obtained in patients with IA rupture compared to unruptured controls. The results of KEGG and GO enrichment analysis showed that neutrophil activation is involved in immune response, neutrophil mediated immunity, and positive regulation of angiogenesis. Interestingly, the results of immunoassays demonstrated that the break in IA may be associated with immune T cells. We used DEGs and WGCNA to determine common genes. The logistic regression model was trained based on 24 intersecting genes, and eventually retained 2 genes, KIAA0226L and UPP1, which we found to be reliable using the validation set, and GSEA revealed that the diagnostic model was associated with the Hippo signaling pathway and vascular smooth muscle contraction, and viral protein interaction with cytokine and cytokine were also associated. Finally explored using the CMap database, Tivozanib could be a potential small molecule drug for the treatment of ruptured intracranial aneurysms (RIAs). Conclusions: We identified new diagnostic genes associated with IA rupture, which may provide a new way of aneurysm diagnosis.

Effects of the application timing of anti-erosive agents on dentin erosion.

This in vitro study aimed to evaluate the effects of the application timing of anti-erosive agents on dentin erosion. Eighty dentin specimens with dimensions of 2 × 2 × 2 mm were prepared and randomly divided into 4 groups based on the treatment solutions: 1.23 × 104 µg/ml sodium fluoride (NaF), 120 µg/ml chlorhexidine (CHX), 300 µg/ml quercetin (QUE), and deionized water (DW, negative control). The specimens in each group were further divided into 2 subgroups according to the application timing of the treatment solutions (n = 10): before the erosive challenges (PRE) and after the erosive challenges (POST). All specimens were submitted to 4 daily erosive challenges for 5 d. For each erosive challenge, the specimens in the subgroup PRE were treated with the respective solutions for 2 min and then immersed in cola drinks for 5 min, while the specimens in the subgroup POST were immersed in cola drinks for 5 min followed by treatment with the respective solutions for 2 min. The erosive dentin loss (EDL) was measured using a contact profilometer, and the surface morphology of the dentin specimens was evaluated by scanning electron microscopy at the end of the experiment. The data were statistically analyzed using two-way analysis of variance (ANOVA) and Bonferroni's test (α = 0.05). Significantly less EDL was observed in the groups NaF, CHX, and QUE than in the group DW (all P < 0.001). Significantly lower EDL was observed in the groups CHX and QUE than in the group NaF (P = 0.001 and P < 0.001, respectively). For CHX, subgroup POST exhibited significantly less EDL than subgroup PRE (P < 0.001). Regarding QUE, subgroup PRE showed significantly less EDL than subgroup POST (P < 0.001). Furthermore, a relatively greater number of obliterated dentinal tubules was visible in the subgroup POST rather than in the subgroup PRE of the group CHX, while in the group QUE, narrower dentinal tubules were observed in the subgroup PRE than those in subgroup POST. In conclusion, CHX and QUE showed the best performance in controlling dentin erosion. CHX was more effective in reducing EDL when applied after erosive challenges, whereas QUE worked more effectively when used before erosive attacks. The application timing should be considered when evaluating the effects of anti-erosive agents because it may determine their effectiveness.

Network pharmacological analysis of active components of Xiaoliu decoction in the treatment of glioblastoma multiforme.

Background: Glioblastoma multiforme (GBM) is the most aggressive primary nervous system brain tumor. There is still a lack of effective methods to control its progression and recurrence in clinical treatment. It is clinically found that Xiaoliu Decoction (XLD) has the effect of treating brain tumors and preventing tumor recurrence. However, its mechanism is still unclear. Methods: Search the Traditional Chinese Medicine System Pharmacology Database (TCSMP) for efficient substances for the treatment of XLD in the treatment of GBM, and target the targeted genes of the effective ingredients to construct a network. At the same time, download GBM-related gene expression data from the TCGA and GTEX databases, screen differential expression bases, and establish a drug target disease network. Through bioinformatics analysis, the target genes and shared genes of the selected Chinese medicines are analyzed. Finally, molecular docking was performed to further clarify the possibility of XLD in multiple GBMs. Results: We screened 894 differentially expressed genes in GBM, 230 XLD active ingredients and 169 predicted targets of its active compounds, of which 19 target genes are related to the differential expression of GBM. Bioinformatics analysis shows that these targets are closely related to cell proliferation, cell cycle regulation, and DNA synthesis. Finally, through molecular docking, it was further confirmed that Tanshinone IIA, the active ingredient of XLD, was tightly bound to key proteins. Conclusion: To sum up, the results of this study suggest that the mechanism of XLD in the treatment of GBM involves multiple targets and signal pathways related to tumorigenesis and development. This study not only provides a new theoretical basis for the treatment of glioblastoma multiforme with traditional Chinese medicine, but also provides a new idea for the research and development of targeted drugs for the treatment of glioblastoma multiforme.

Network pharmacological analysis of active components of Tongqiao Huoxue Decoction in the treatment of intracerebral hemorrhage.

Background: Intracerebral hemorrhage (ICH) is a type of stroke which results in a high disability and mortality rate and has a poor prognosis. Tongqiao Huoxue Decoction (TQHXD) is a classical Chinese prescription. Clinical practice has proven that TQHXD can promote blood circulation and can effectively treat ICH and its sequelae. However, the current mechanism is still unclear. Methods: The chemical components and target genes of TQHXD were collected from the Traditional Chinese medicine (TCM) Systems Pharmacology and Bioinformatics Analysis Tool for Molecular Mechanism of Traditional Chinese Medicine analysis platforms, and the gene expression data of ICH tissues were downloaded from the Gene Expression Omnibus (GEO) database. Weighted gene co-expression network analysis (WGCNA) was performed to obtain differentially co-expressed gene pairs and build a drug-target-disease network. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were performed on the obtained target genes and shared genes. Finally, molecular docking was carried out to further clarify the utility of TQHXD for the treatment of ICH. Results: A total of 304 differentially expressed genes in ICH, 42 TQHXD active ingredients, and 279 predicted targets of its active compounds were obtained. Bioinformatics analysis showed that they were involved in angiogenesis, the regulation of wound healing, and other biological processes. Furthermore, their participation in fluid shear stress and the atherosclerosis signaling pathway indicated their close association with the pathological processes of ICH. Finally, molecular docking was carried out to further confirm the tightly binding structural sites of the effective components of TQHXD and key proteins. Conclusions: In summary, the results of this study suggest that the mechanism of action of TQHXD in the treatment of ICH involves multiple targets and signaling pathways related to its occurrence and development. This study not only provides a new theoretical basis for the treatment of ICH with traditional Chinese medicine, but also provides new ideas for the research and development of drugs for the treatment of ICH.

[Small interfering RNA-mediated α-enolase knockdown suppresses glycolysis and proliferation of human glioma U251 cells in vitro].

OBJECTIVE: To investigate the role of α-enolase (ENO1) in regulating glucose metabolism and cell growth in human glioma cells. METHODS: Glucose uptake and lactate generation were assessed to evaluate the changes in glucose metabolism in human glioma U251 cells with small interfering RNA (siRNA)-mediated ENO1 knockdown. MTT assay and 5-ethynyl-2'-deoxyuridine (EdU) staining were used to examine the cell growth and cell cycle changes following siRNA transfection of the cells. RESULTS: Transfection of U251 cells with siRNA-ENO1 markedly reduced glucose uptake (P=0.023) and lactate generation (P=0.007) in the cells and resulted in significant suppression of cell proliferation (*P<0.05) since the second day following the transfection. Transfection with siRNA-ENO1 also obviously suppressed cell cycle G1/S transition in the cells (P=0.0425). The expressions of HK2 and LDHA, the marker genes for glucose metabolism, were significantly down-regulated in the cells with siRNA-mediated ENO1 knockdown. CONCLUSION: ENO1 as a potential oncogene promotes glioma cell growth by positively modulating glucose metabolism.

[Pathological analysis and mRNA expression of apoptosis genes in rat kidney tissue after subacute melamine treatment].

OBJECTIVE: to investigate pathological changes and mRNA expression of apoptosis genes bcl-2, bax and Caspase-3 in rat kidney tissue after rats are administrated with melamine for 28 days. METHODS: 10 male SD rats and 10 female SD rats in each group were administrated with three doses of melamine (low dose, middle dose, high dose) by gavage for 28 days. The animals were divided into three experimental groups and one control group. The doses for male rats were 200, 400, 800 mg/kg, but for female rats they were 150, 300, 600 mg/kg. After melamine treatment the animals were sacrificed and the kidneys were taken out for pathological analysis and for detecting mRNA expression of bcl-2, bax and Caspase-3 with fluorescent quantitative PCR assay. RESULTS: the tubular cylinders were observed in three experimental groups. The positive rates of tubular cylinders in three groups (from low dose to high dose) were 11/20, 13/20, 16/20, respectively. Additionally, melamine induced a significant decrease in mRNA expression of bcl-2 at low dose, middle dose or high dose, bcl-2 expression decreased by 20.58% - 49.51% in three groups treated with melamine. Furthermore, bax mRNA levels increased by 44.66% - 300.96% in groups treated with three doses of melamine, and Caspase-3 mRNA levels increased by 64.76% - 360.75% in groups treated with three doses of melamine. CONCLUSIONS: melamine could induce pathological changes of rat kidneys, and it also induces a significant alteration of apoptosis Bcl-2, Bax and Caspase-3 mRNA expression in rat kidney tissue.

[Study on skin sensitization as well as liver and kidney impairment in guinea pigs treated with trichloroethylene].

OBJECTIVE: To study skin sensitization as well as liver and kidney impairment in guinea pigs treated with trichloroethylene (TCE). METHODS: Guinea pig maximization test (GPMT) was applied in this study, guinea pigs were divided into 3 groups, namely negative control, positive control and TCE treatment. Animals of 3 groups were administrated with olive oil, 2, 4-dinitrochlorobenzene (DNCB), and TCE, respectively, by intradermal injection. The animal skin was observed and blood was collected after various treatment, the liver function tests were conducted, including detection of activities of ALT, AST, LDH and levels of creatinine, uric acid, and urea with automatic biochemical analyzer. RESULTS: Obvious skin impairment was observed in the groups of positive control and TCE treatment, the skin impairment included erythema and edema, the sensitization rate was 100% in positive control and 83.3% in TCE treatment group. Additionally, the activities of ALT, AST and LDH increased significantly in the groups of positive control and TCE treatment when compared with the negative control. CONCLUSIONS: Trichloroethylene is one of the strong hypersensitizing substances, it could induce skin allergic reaction and liver impairment in guinea pigs.

[Effect of cadmium chloride on the expression and phosphorylation of mitogen-activated protein kinase in normal rat kidney epithelial cells].

OBJECTIVE: To explore the effect of cadmium chloride on the expression and phosphorylation of mitogen-activated protein kinase (MAPK) in normal rat kidney epithelial (NRK) cells. METHODS: The NRK cells were incubated with cadmium chloride either at respective dose (0, 1, 5, 10, 20, 40 µmol/L) for 24 h or at same dose (10 µmol/L) for respective time (0, 0.5, 1.0, 2.0, 4.0, 8.0 h). Western blotting was applied to test the expression of MAPK in NRK cells (ERK1/2, p38, JNK); and phosphor-specific antibody to detect the phosphorylated MAPK (p-ERK1/2, p-p38, p-JNK). RESULTS: There was no significant difference in the MAPK expression among the groups either treated with different doses or for different time; however, the level of phosphorylated MAPK was comparatively higher than it in control group. There was an obvious expression of p-ERK1/2 at 1.00 ± 0.06 in the group incubated with 10 µmol/L CdCl(2); and the expression in the 20 µmol/L and 40 µmol/L CdCl(2) group was 2.58 ± 0.11, 2.76 ± 0.23 respectively, which was 1.58 and 1.76 times more than it in 10 µmol/L CdCl(2) group. The differences were statistically significant (F = 827.70, P < 0.01). The respective expression of p-p38MAPK in the 20 µmol/L (2.47 ± 0.20)and 40 µmol/L CdCl(2) group (3.73 ± 0.25)was 1.47 and 2.73 times more than it in control group (1.00 ± 0.02), and the differences were also statistically significant (F = 280.06, P < 0.01). There was a dose-effect relationship of the concentration of cadmium in the expression of p-ERK1/2 (r = 0.919, t = 4.69, P = 0.009) and p-p38MAPK (r = 0.945, t = 5.79, P = 0.004). Additionally, phosphorylated MAPK expressed in a time-dependent manner. The expression of p-ERK1/2 was obvious in the group incubated for 1 h (1.26 ± 0.11), and the respective expression in the 4 h group (1.51 ± 0.07) and 8 h group (3.53 ± 0.23) was 1.5 and 3.5 times of it in the control group (1.00 ± 0.02). The differences were statistically significant (F = 427.82, P < 0.001). The expression of p-p38MAPK increased significantly in 1 h group (1.31 ± 0.07); while the respective expression in 4 h group (3.53 ± 0.32) and 8 h group (4.41 ± 0.38) was 3.5 and 4.4 times of it in control group (1.00 ± 0.03). The differences were also statistically significant (F = 280.06, P < 0.001). CONCLUSION: Cadmium chloride could significantly enhance the phosphorylation of MAPK in NRK cells; and it is probably associated with the activation of MAPK.

Up-regulating PPAR-γ expression and NO concentration, and down-regulating PAI-1 concentration in a rabbit atherosclerotic model: the possible antiatherogenic and antithrombotic effects of atorvastatin.

We investigated the effect of atorvastatin on the plasma concentration of plasminogen activator inhibitor-1 (PAI-1) and nitric oxide (NO) in a rabbit model, and the relationship between these effects and peroxisome proliferator-activated receptor γ (PPAR-γ). In our experiments, 24 male Japanese rabbits were divided into 3 groups: the high-cholesterol diet group (the high-C group), the high-cholesterol diet plus atorvastatin group (the atorvastatin group), and the normal diet group (the control group). All rabbits were killed after a 16-week feeding. The expression of PPAR-γ and the plasma concentrations of NO and PAI-1 were evaluated by an immunohistochemical assay while the level of the plasma lipid profile was measured using a commercially available kit. The atorvastatin not only reduces the plasma levels of the total cholesterol (TC) and the low-density lipoprotein cholesterol (LDL-C), but also increases the expression of PPAR-γ and the concentration of NO in comparison to the control group [16.11 ± 2.35% vs 7.68 ± 1.04%; 249.30 ± 27.90 vs 179.12 ± 28.51 (µml/L), p<0.05 respectively]. In addition, the concentration of PAI-1 in the atorvastatin group is lower than that in the control group (0.11 ± 0.01A vs 0.14 ± 0.02A, p<0.05). The changes of PAI-1 and NO in the atorvastatin group are in good accordance to that of PPAR-γ. Results show that atorvastatin significantly up-regulates the expression of nuclear transcription factor, namely PPAR-γ, and induces the changes of the other two factors, which might provide mechanisms for the antiatherosclerotic and antithrombotic effects of atorvastatin.