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Objective To investigate the effect of calcium ions on the expression of kallikrein-4 (KLK4) and cell growth of ameloblast, and to provide an experimental basis for calcium ion promoting normal mineralization of enamel. Methods ALC cells were treated with 0, 2.0, 2.5, 3.0, and 3.5 mmol/L CaCl2 for 24 and 48 h. KLK4 expression was analyzed by qRT-PCR and Western blot analysis. The viability of ALC cells was determined by using CCK-8. AnnexinV-FITC/PI dual staining combined with flow cytometry and Hoechst 33342 staining were used to detect the ALC cell cycle and cell apoptosis. The protein expression level of glucose-regulated protein 78 (GRP78) was measured by Western blot analysis. Results After 24 h of treatment with 2.5, 3.0, and 3.5 mmol/L CaCl2, the expression of KLK4 mRNA was increased (P2, the expression of KLK4 protein was increased (P2, the expression of KLK4 mRNA and protein was increased (P2 (P2. Hoechst 33342 staining results showed that 3.0 mmol/L and 3.5 mmol/L CaCl2 may promote apoptosis in ALC cells. Flow cytometry showed that the proportion of G2/M phase cells and the apoptosis rate increased after 3.5 mmol /L CaCl2 treatment for 24 h (P2 groups. After 24 h of treatment with 3.0 mmol/L and 3.5 mmol/L CaCl2, the expression of GRP78 protein was reduced (P2, the expression of GRP78 protein was reduced (P

Copyright of Journal of Prevention & Treatment For Stomatological Diseases is the property of Journal of Prevention & Treatment For Stomatological Diseases Editorial Office and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Objectives To construct human breast cancer cell lines HS578T and MDA-MB-231 that overexpress the endoplasmic reticulum stress marker glucose-regulated protein 78（GRP78), and to provide a basis for exploring the mechanism of GRP78 in the development and progression of breast cancer. Methods The lentiviral vector pCDH-CMVGRP78-HA-EF1-Puro plasmid expressing GRP78 was constructed by molecular cloning technology, and the plasmid was amplified by PCR method. Then the plasmid was cut by EcoRⅠ and NotⅠ restriction enzymes, and the digestion products were identified by sequencing. The pCDH-CMV-GRP78-HA-EF1-Puro plasmid was constructed successfully. 293FT cell line （clonal isolate） in the logarithmic growth phase was added into a culture dish with pCDH-CMV-GRP78-HA-EF1-Puro mixture, and was cultured for 48 h； the supernate containing GRP78 lentiviral particles was collected and stored for later use. In addition, human breast cancer cell lines HS578T and MDA-MB-231 in the logarithmic growth phase were cultured in the medium containing supernate of GRP78 lentiviral particles. After 48 h culture, 1：500 diluted purinycin was added to the medium, and the cell lines of HS578T and MDA-MB-231 with overexpression of GRP78 were screened. The GRP78 of HS578T and MDA-MB-231 cells was detected by Western blotting after being cultured with purinamycin for 48 h. The GRP78 protein in HS578T and MDA-MB-231 cells was localized by confocal laser microscopy. Results The cell lines of HS578T and MDA-MB-231 with overexpression of GRP78 showed obvious GPR78 protein electrophoretic bands around 78 kd. There was GRP78 protein expression in HS578T cells, which was mainly distributed in the cytoplasm. Conclusions We successfully constructed MDA-MB-231 and HS578T cell lines with GRP78 overexpression, and GRP78 was mainly ex‐ pressed in the cytoplasm. [ABSTRACT FROM AUTHOR]

Objective To study the effect of low concentrations of sodium fluoride on the osteogenic/odontogenic differentiation of human dental pulp cells (hDPCs) in vitro. Methods This study was reviewed and approved by the Ethics Committee. hDPCs were cultured using a modified tissue explant technique in vitro. The effects of different concentrations of sodium fluoride on the proliferation of hDPCs were measured by methylthiazol tetrazolium (MTT) assay. Appropriate concentrations were added to the osteogenic/odontogenic differentiation induction medium, and the cells were induced in vitro. Alizarin red S staining was used to detect the osteoblastic/odontogenic differentiation ability of the cells, and the mRNA expression of the key differentiation factors was detected by RT-qPCR. Moreover, the expression of key molecules of endoplasmic reticulum stress (ERS) was detected by RT-qPCR and Western blot. The data were analyzed with the SPSS 18.0 software package. Results Low concentration of NaF (0.1 mmol/L) could stimulate cell proliferation in vitro, while a high concentration (5-10 mmol/L) could inhibit cell proliferation (P

Copyright of Journal of Prevention & Treatment For Stomatological Diseases is the property of Journal of Prevention & Treatment For Stomatological Diseases Editorial Office and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Acta Agriculturae Zhejiangensis is the property of Acta Agriculturae Zhejiangensis Editorial Office and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

To investigate the effects of quercetin on oxidative stress and endoplasmic retic‑ ulum stress of human umbilical vein endothelial cells（HUVECs）induced by high glucose and their pos‑ sible mechanism. Methods HUVECs were cultured in vitro，and the cells were divided into the control group（Control group），high glucose（HG）group，and quercetin groups with different concentrations （20 µmol/L，50 µmol/L，and 100 µmol/L，named as Q20，Q50，Q100 groups），respectively. HU‑ VECs in the Q20 group were pretreated with 20 µmol/L quercetin for 1 h，and then were added with 10 mg/L glucose for culture. HUVECs in the Q50 group were pretreated with 50 µmol/L quercetin for 1 h，and then were added with glucose for culture. HUVECs in the Q100 group were pretreated with 100 µmol/L quercetin for 1 h，and then were added with 10 mg/L glucose for culture. In the HG group， 10 mg/L HG was added to the culture medium，and the cells in the Control group were cultured in the low-glucose DMEM medium. After 24 hours of intervention，the cell viability was detected by cell count‑ ing Kit-8（CCK-8）method. The apoptosis rate and reactive oxygen species（ROS）level were detected by flow cytometry. The degree of cell membrane damage was evaluated by detecting lactate dehydrogenase （LDH）activity in the media. Western blotting was used to detect the expression levels of glucose-regulat‑ ed protein 78 （GRP78），CCAAT/enhancer-binding protein homologous protein （CHOP） and Cas‑ pase12. Results Compared with the Control group，the activity of endothelial cells in the HG group de‑ creased after 24 h（P<0. 05）. Compared with the HG group，the activity of endothelial cells in the Q20， Q50 and Q100 groups increased，and the cell viability increased with the increased concentrations of quercetin（all P<0. 05）. Compared with the Control group，the apoptosis rate of the HG group increased （P<0. 05），while the apoptosis rates of the Q20，Q50 and Q100 groups decreased，and the apoptosis rate decreased with the increased concentrations of quercetin（all P<0. 05）. Compared with the Control group，the ROS in the HG group increased（P<0. 05）；compared with the HG group，the ROS in the Q20，Q50 and Q100 groups decreased，and the ROS decreased with the increased concentrations of quer‑ cetin（all P<0. 05）. Compared with the Control group，the LDH activity in the cell culture medium of the HG group increased（P<0. 05）. Compared with the HG group，the LDH activity in the cell culture medi‑ um of the Q50 group and Q100 group decreased （both P<0. 05）. Compared with the Q50 group， the LDH activity in the cell culture medium of the Q100 group decreased（P<0. 05）. Compared with the Control group，the expression levels of GRP78，CHOP，and Caspase 12 in the endothelial cells of the HG group increased（all P<0. 01）. Compared with the HG group，the expression levels of GRP78， CHOP，and Caspase 12 decreased in the endothelial cells of the Q20，Q50 and Q100 groups（all P< 0. 01）. Conclusions Quercetin can increase endothelial cell activity，reduce LDH activity，inhibit the apoptosis，reduce ROS content，and inhibit the expression levels of GRP78，CHOP and Caspase 12 of endothelial cells in the high glucose solution，and its inhibitory effect is enhanced with the increased con‑ centrations of quercetin. Quercetin may play a protective effect on endothelial cells by inhibiting endothe‑ lial cell oxidative stress，endoplasmic reticulum stress and reducing apoptosis under the condition of high glucose. [ABSTRACT FROM AUTHOR]

Objective: To analyze the clinical application value of serum heme oxygenase (HO)-1expression level in non-alcoholic fatty liver disease (NAFLD) and, based on that, establish a diagnostic model combined with glucose regulatory protein 78 (GRP78) so as to clarify its diagnostic effectiveness and application value. Methods: A total of 210 NAFLD patients diagnosed by abdominal B-ultrasound and liver elastography were included, and at the same time, 170 healthy controls were enrolled. The general clinical data, peripheral blood cell counts, and biochemical indicators of the research subjects were collected. The expression levels of HO-1 and GRP78 were detected using an enzyme-linked immunosorbent assay. Multivariate analysis was used to screen independent risk factors for NAFLD. Visual output was performed through nomogram diagrams, and the diagnostic model was constructed. Receiver operating characteristic curve (ROC), calibration curve, and decision curve analysis (DCA) were used to evaluate the diagnostic effectiveness of NAFLD. Measurement data were analyzed using a t -test or Mann-Whitney U rank sum test to detect data differences between groups. Enumeration data were analyzed using the Fisher's exact probability test or the Pearson χ (2) test. Results: Compared with the healthy control group, the white blood cell count, aspartate aminotransferase (AST), alanine aminotransferase, gamma-glutamyl transferase (GTT), fasting blood glucose (Glu), triglyceride (TG), low-density lipoprotein cholesterol (LDL-C), serum HO-1, and GRP78 levels were significantly increased in the NAFLD group patients ( P  < 0.05). Binary logistic analysis results showed that AST, TG, LDL-C, serum HO-1, and GRP78 were independent risk factors for NAFLD ( P  < 0.05). A nomogram clinical predictive model HGATL was established using HO-1 (H), GRP78 (G) combined with AST (A), TG (T), and LDL-C (L), with the formula P =-21.469+3.621×HO-1+0.116 ×GRP78+0.674×AST+6.250×TG+4.122 ×LDL-C. The results confirmed that the area under the ROC curve of the HGATL model was 0.965 8, with an optimal cutoff value of 81.69, a sensitivity of 87.06%, a specificity of 92.82%, a P  < 0.05, and the diagnostic effectiveness significantly higher than that of a single indicator. The calibration curve and DCA both showed that the model had good diagnostic performance. Conclusion: The HGATL model can be used as a novel, non-invasive diagnosis model for NAFLD and has a positive application value in NAFLD diagnosis and therapeutic effect evaluation. Therefore, it should be explored and promoted in clinical applications.

Objective To explore the effect of drug-containing serum of Zhishi (immature hitter orange) on endoplasmic reticulum stress (ERS) injury in rat gastric interstitial cells of Cajal (ICCs). Methods ICCs were isolated by collagenase digestion and identified by immunofluorescence staining. Tunicamycin (Tm) was used to induce ICCs to establish ERS models, the 4-Phenylbutyric acid (4-PBA) was used to block autophagy. Rat drug-containing serum of Zhishi was prepared by using the serum pharmacology routine method. ICCs were randomly divided into four groups after subculture, including the blank group (normal culture), ERS group (the ICCs were stimulated with 1 μg /mL Tm), 4-PBA group (the ICCs were stimulated with 1 μg /mL Tm and 10 mmol/L 4-PBA), and Zhsihi group (the ICCs were stimulated with 1 μg /mL Tm and 20% drug-containing serum), and we continued to cultivate ICCs for 24 h. The ultrastructure of endoplasmic reticulum in ICCs was observed by transmission electron microscopy. The mRNA expression levels of GRP78 and ATF6 were detected by real-time fluorescent quantitative PCR. Results We isolated and identified ICCs successfully. Under transmission electron microscope, we observed that the endoplasmic reticulum of ICCs was normal and well-arranged in the blank group; the number of organelles reduced, and endoplasmic reticulum swelling, dilation, vesiculation changes and disordered arrangement were found in the ERS group; ICCs had endoplasmic reticulum swelling and limitation expansion in the Zhsihi group. Compared with the ICCs group, the mRNA expression of GRP78 and ATF6 increased in the ERS group (both P <0.05). Compared with the ERS group, the mRNA expression of GRP78 and ATF6 decreased in the Zhishi group (both P < 0.05). Conclusion The drug-containing serum of Zhishi can reduce the ERS injury of ICCs possibly by regula-ting the ATF6 signaling pathway of ERS. [ABSTRACT FROM AUTHOR]

Objective To explore the effect of curcumin on endoplasmic reticulum stress of alveolar epithelial cells in a rat model of chronic obstructive pulmonary disease (COPD). Methods Male SD rats were randomly divided into normal control, COPD model, curcumin-treated and solvent control groups. The rat model of COPD was induced by cigarette smoke exposure combined with intratracheal administration of lipopolysaccharide, and rats were further treated according to different groups. The numbers of total cells, neutrophils and macrophages in bronchoalveolar lavage fluid (BALF) were counted. Pathological changes of lung tissues were studied by HE staining. The endoplasmic reticulum morphology of alveolar epithelial cells was observed using transmission electron microscopy. The protein expression of glucose-regulated protein 78 (GRP78) in alveolar epithelial cells was detected by immunohistochemistry. Results Compared with those in normal control group, the numbers of total cells, neutrophils and macrophages in BALF from rats in COPD model group were all significantly increased (P<0.05). The numbers of total cells, neutrophils and macrophages in BALF from rats in curcumin-treated group were significantly lower than those in COPD model group (P<0.05). Light microscopy and transmission electron microscopy showed enlarged alveolar space and swollen endoplasmic reticulum of alveolar epithelial cells in rats from COPD model group. The damage of alveolar epithelial cells and endoplasmic reticulum in COPD rats was attenuated by curcumin treatment. Compared with that in normal control group, the expression of GRP78 in alveolar epithelial cells in rats from COPD model group was significantly increased (P<0.05). And the protein expression level of GRP78 in alveolar epithelial cells in COPD rats was significantly down-regulated by curcumin treatment (P<0.05). Conclusion Curcumin plays a protective effect against COPD possibly by inhibiting endoplasmic reticulum stress of alveolar epithelial cells. [ABSTRACT FROM AUTHOR]

Copyright of Journal of Capital Institute of Physical Education is the property of Shoudu Tiyu Xueyuan and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)