OBJECTIVETo determine the effect of type I transmembrane protein (IRE1alpha) deletions on the cell cycle of human periodontal ligament fibroblasts (hPDLFs) cells.

METHODSBased on the IRE1alpha deletions, a full-length model was successfully constructed. Moreover, overlapping polymerase chain reaction mutagenesis facilitated the establishment of two deletion mutants of IREla (pD-Kinase, pD-Rnase). The full-length model and two mutant eukaryotic expression vectors were transfected into hPDLFs cells. Western blot analysis was performed to identify the expression in the cells. The changes in the cell cycle of hPDLFS cells were detected by flow cytometry (FCM).

RESULTSThe two deletion mutants of IRE1alpha with eukaryotic expression vectors were successfully constructed and correctly expressed in hPDLFs cells based on Western blot analysis. Under stress conditions, the FCM assay showed that cell percentage of S phases increased, whereas that of G1 phases decreased in the IRE1alpha group (P < 0.05) compared with the control group of tunicamycin (TM) treatment. Moreover, the cell percentage of the S phases decreased, whereas that of the G1 phases increased in the D-Rnase group (P < 0.05) compared with the control. The deletion mutant D-Kinase had no influence on hPDLFS cell proliferation and cycle (P>0.05).

CONCLUSIONUnder stress conditions, IRE1alpha can improve the cell cycle of hPDLFs cells from the G1 to the S phase. The deletion mutant D-Rnase cause hPDLFs cell growth arrest at the G1 phase, whereas deletion mutant D-Kinase has no significant effect.

Objective To evaluate the effects of propolis on the growth of S.mutans ATCC 25175(S.m),S.sobrinus 6715(S.s) and their fluoride-resistant strains(S.m-FR,S.s-FR),and the change of their glucosyltransferase(GTF).Methods S.m and S.s were induced with sodium fluoride by minimum inhibitory concentration(MIC) to induce fluoride-resistant strains(S.m-FR and S.s-FR).Fluid dilution method was used to observe the effects of different levels of propolis on growth of S.m,S.m-FR,S.s and S.s-FR.Chemistry enzyme analysis was used to detect the influence of propolis on the enzyme activity of GTF.Results MIC of propolis for S.m,S.m-FR,S.s and S.s-FR were respectively 0.39,0.78,0.20 and 0.39 g/L.MBC of propolis for them were respectively 0.78,1.56,1.56,and 1.56 g/L.GTF of S.m,S.m-FR,S.s,and S.s-FR was decreased gradually with the increase of concentration of propolis.There were significant differences among total sample groups,and each experimental group and positive control group as well(P

Objective To study the intracellular localization and temporal expression of CPSIT_0271 in Chlamydia psittaci-infected cells; and to investigate the effects of recombinant GST-CPSIT_0271 protein on the expression of proinflammatory cytokines including IL-6, IL-1βand TNF-αby THP-1 cells.Methods The gene encoding CPSIT_0271 of Chlamydia psittaci was expressed as fusion protein ( GST-CPSIT_0271 ) in E.coli.The polyclonal antibody was prepared by immunizing BALB /c mice with the purified recombinant pro-tein.Antibody titer was determined by ELISA .Indirect immunofluorescence assay ( IFA) was performed to lo-cate the endogenous CPSIT_0271 protein in C.psittaci-infected cells .The expression characteristics of CPSIT_0271 protein were detected by Western blot in C.psittaci-infected HeLa cells at different time points .The levels of IL-6, IL-1βand TNF-αwere analyzed by ELISA after stimulating THP-1 cells with different concentrations of CPSIT_0271 protein.Results CPSIT_0271 protein was found to express in the chlamydia inclusion of C.psittaci-infected HeLa cells .The expression of CPSIT_0271 protein was detected firstly at 36 h and increased at 48 h after C.psittaci infection.The titer of anti-CPSIT_0271 specific antibody in GST-CPSIT_0271 immu-nized mice reached to 1 ∶16 000.GST-CPSIT_0271 protein increased the levels of IL-6, IL-1βand TNF-αin THP-1 cells in a dose-dependent manner in the range of 2 to 5 μg/ml.The levels of TNF-αand IL-1βreached their peaks at 24 h, and IL-6 level peaked at 48 h upon the stimulation by 5 μg/ml of GST-CPSIT_0271 pro-tein.Conclusion CPSIT_0271 expressed in inclusion bodies of Chlamydia psittaci in the infected cells , sug-gesting it might be a late expression gene .GST-CPSIT_0271 protein shows good immunogenicity and enhances the expressions of IL-6, IL-1βand TNF-αin THP-1 cells.

Objective To observe the degradation time and the intimal hyperplasia of biodegradable magnesium alloy stent (MPM) implanted in the abdominal aorta of experimental rabbits.Methods A total of 24 New Zealand white rabbits were randomly divided into four groups (30 d,60 d,90 d and 180 d) with 6 rabbits in each group.In cach rabbit one MPM stent was implanted in the abdominal aorta at the level of one cm below the left renal artery.Reexamination of abdominal aortography with DSA was separately performed at 30,60,90 and 180 d after stent implantation to check the stent condition.The rabbits of each group were sacrificed at the corresponding scheduled day,the stenting segment of aorta of each rabbit was removed and the specimen was sent for microscopic examination.The experimental results were analyzed with SPSS20.0 software.Results All the 24 experimental rabbits survived.During the follow-up period the stent showed gradual degradation changes,and basically complete degradation was not observed until to 180 days.Meanwhile,the intimal hyperplasia reached its peak at 90 days after implantation.The abdominal aorta remained unobstructed during the whole process of degradation.Conclusion The time of complete degradation for MPM stent is 182 days,which is long enough to meet the needs of vascular positive remodeling.

Objective To discuss the application of different types of bronchial arteriography catheter in performing bronchial artery embolization (BAE) for the treatment of hemoptysis.Methods The clinical data of a total of 97 patients with hemoptysis,who received BAE during the period from January 2013 to May 2016,were collected.According to angiographic findings in aspect of the opening and running direction of the arteries causing bleeding,the responsible arteries were divided into 4 types:upward opening,horizontal opening and running upwards,horizontal opening and running downwards,and downward opening.For each responsible artery,appropriate angiography catheter was selected from the following catheters:MIK catheter,left gastric artery catheter,Cobra catheter,Simmon-1 catheter and Simmon-2 catheter.With super-selective catheterization technique the selected suitable catheter was inserted into the responsible artery and angiography was subsequently performed.The effect of the selection of bronchial arteriography catheter in performing BAE for hemoptysis was analyzed.Results A total of 180 responsible arteries were detected in 97 patients.Of the 180 responsible arteries,artery with upward opening was seen in 42,artery with horizontal opening and running upwards was found in 54,artery with horizontal opening and running downwards was observed in 46,and artery with downward opening was detected in 38.The success rates of super-selective catheterization for MIK catheter,left gastric artery catheter,Cobra catheter and Simmon catheter were 83.3％ (35/42),92.6％ (50/54),87.0％ (40/46) and 89.5％ (34/38,including 30 Simmon-1 catheters and 4 Simmon-2 catheters) respectively.After BAE,the responsible arteries were occluded in all patients,and hemoptysis stopped immediately.The recurrence rate at 6 months after BAE was 7.2％ (7/97).Conclusion For the treatment of hemoptysis,BAE is safe and effective.The key point to ensure a successful BAE is that the selection of appropriate catheter should be based on the opening and running direction of the artery causing bleeding.

Objective To investigate the effects of dexamethasone on absorption of lung edema in rabbits with seawater drowning-induced acute lung injury.Methods Seawater(4 ml/kg body weight)was instilled into the lower trachea of ventilated and anesthetized rabbits.These rabbits were assigned randomly to receive intravenous injection of 1 mg/ kg body weight of dexamethasone(dexamethasone group,DG)or 2 ml of normal saline(control group,CG).Lung edema was measured by extravascular lung water index(EVLWI)using a gravimetric method.Three hours after treatment, epithelial Na~+ channel subunit-?(?-ENaC)mRNA and Na~+/K~+-adenosine triphosphatase subunit-?l(NKA-?l) protein abundances in lung tissues were respectively measured by reverse transcriptase-polymerase chain reaction and Western blotting,and NKA activity was measured by monitoring the release of inorganic phosphate(Pi)from adenosine triphosphate(ATP).Results The DG's EVLWI was significantly lower than the CG's[(0.508?0.089)vs.(0.648?0.102),P＜0.05)],but the DG's NKA activity,?-ENaC mRNA and NKA-?l protein abundances were significantly higher than the CG's,correspondingly(P＜0.05).Conclusions With up-regulation of the NKA activity and expressions of?-ENaC and NKA-?l,dexamethasone treatment could promote the absorption of lung edema in rabbits with seawater drowing-induced acute lung injury.

In the process of conducting implementation research on health service issues, stakeholders' preference for contents related to evidence-based practice (EBP) and implementation strategies is closely related to whether EBP can be effectively implemented.However, multiple preference assessment methods exist, each with their own strengths, weaknesses, and application scenarios, which makes it challenging for researchers to select appropriate and effective preference assessment methods. This paper aims to review the origins, characteristics, and application scenarios of commonly used preference assessment methods, with the hope of providing valuable reference and lessons for domestic scholars to select and apply appropriate preference assessment methods in implementation research.

0.05).Myocardial perfusion defect scores were decreased significantly from 14.8?3.0 to 10.5?1.8(P

Purpose: Temozolomide is used in first-line treatment for glioblastoma. However, chemoresistance to temozolomide is common in glioma patients. In addition, mechanisms for the anti-tumor effects of temozolomide are largely unknown. Ferroptosis is a form of programmed cell death triggered by disturbed redox homeostasis, overloaded iron, and increased lipid peroxidation. The present study was performed to elucidate the involvement of ferroptosis in the anti-tumor mechanisms of temozolomide. Materials and Methods: We utilized the CCK8 assay to evaluate cytotoxicity. Levels of lactate dehydrogenase (LDH), malondialdehyde (MDA), iron, and glutathione (GSH) were measured. Flow cytometry and fluorescence microscope were used to detect the production of reactive oxygen species (ROS). Western blotting, RT-PCR and siRNA transfection were used to investigate molecular mechanisms. Results: Temozolomide increased the levels of LDH, MDA, and iron and reduced GSH levels in TG905 cells. Furthermore, we found that ROS levels and DMT1 expression were elevated in TG905 cells treated with temozolomide and were accompanied by a decrease in the expression of glutathione peroxidase 4, indicating an iron-dependent cell death, ferroptosis. Our results also showed that temozolomide-induced ferroptosis is associated with regulation of the Nrf2/HO-1 pathway. Conversely, DMT1 knockdown by siRNA evidently blocked temozolomide-induced ferroptosis in TG905 cells. Conclusion Taken together, our findings indicate that temozolomide may suppress cell growth partly by inducing ferroptosis by targeting DMT1 expression in glioblastoma cells.

Purpose: Temozolomide is used in first-line treatment for glioblastoma. However, chemoresistance to temozolomide is common in glioma patients. In addition, mechanisms for the anti-tumor effects of temozolomide are largely unknown. Ferroptosis is a form of programmed cell death triggered by disturbed redox homeostasis, overloaded iron, and increased lipid peroxidation. The present study was performed to elucidate the involvement of ferroptosis in the anti-tumor mechanisms of temozolomide. Materials and Methods: We utilized the CCK8 assay to evaluate cytotoxicity. Levels of lactate dehydrogenase (LDH), malondialdehyde (MDA), iron, and glutathione (GSH) were measured. Flow cytometry and fluorescence microscope were used to detect the production of reactive oxygen species (ROS). Western blotting, RT-PCR and siRNA transfection were used to investigate molecular mechanisms. Results: Temozolomide increased the levels of LDH, MDA, and iron and reduced GSH levels in TG905 cells. Furthermore, we found that ROS levels and DMT1 expression were elevated in TG905 cells treated with temozolomide and were accompanied by a decrease in the expression of glutathione peroxidase 4, indicating an iron-dependent cell death, ferroptosis. Our results also showed that temozolomide-induced ferroptosis is associated with regulation of the Nrf2/HO-1 pathway. Conversely, DMT1 knockdown by siRNA evidently blocked temozolomide-induced ferroptosis in TG905 cells. Conclusion Taken together, our findings indicate that temozolomide may suppress cell growth partly by inducing ferroptosis by targeting DMT1 expression in glioblastoma cells.