Exploring mechanical properties and failure mechanisms of aramid and PBO crystals through molecular dynamics simulations.

Molecular dynamics simulations were used to analyze the mechanical properties and failure processes of poly(p-phenylene-terephthalamide) (PPTA), poly(p-phenylene-benzimidazole-terephthalamide) (PBIA), PBIA-PPTA (formed by 1:1 copolymerization of PPTA and PBIA), and poly(p-phenylene-benzobisoxazole) (PBO) crystals at different strain rates and temperatures. The failure stress and strain were found to be linear with the temperature and logarithmic strain rate. Moreover, based on the kinetic theory of fracture and the comprehensive simulation results, we formulated a model that describes the failure stress of the aforementioned crystals under varying strain rates and temperatures. Through the analysis of the failure process, we found that in PPTA, PBIA, and PBIA-PPTA crystals, the bond failure probability is correlated with the strain rate and temperature. The examination of bond lengths and angles unveiled that bonds with larger initial aligning angles are more susceptible to failure during the strain process. Intriguingly, the stretching process induced a conformational change in the PBO molecular chain, leading to a deviation from the linear relation in its stress-strain curve.

Predictive value of serum ß2-microglobulin in cardiac valve calcification in maintenance hemodialysis patients.

Background: Cardiac valve calcification (CVC) is associated with adverse cardiovascular events. We studied the risk factors of CVC in maintenance hemodialysis (MHD) patients and the value of serum ß2-microglobulin (ß2-MG) levels in predicting the incidence of CVC. ß2-MG is a middle molecular weight toxin. In recent years, researchers found that elevated blood ß2-MG was associated with coronary, thoracic, and abdominal aortic calcifications with significant correlations. ß2-MG has been emerging as a strong biomarker for cardiovascular mortality in uremic patients but its role in CVC is not well studied. This study looked specifically at CVC occurrence in relation to ß2-MG for MHD patients. Methods: Patients who underwent MHD for more than 3 months in the First People's Hospital of Nantong City from November 2012 to November 2019 with complete available data were included in the study. The patients were divided into the CVC group and the non-CVC group. The general information and clinical laboratory indicators of the patients were collected in a retrospective manner. We analyzed the risk factors for developing CVC in MHD patients using binary logistic regression method. Receiver operating characteristic (ROC) curves were used to calculate the cut-off value of ß2-MG for predicting CVC. The decision tree (DT) method was used to classify and explore the probability of CVC in patients with MHD. Results: The ß2-MG in the CVC group was significantly higher than that in the non-CVC group (t=6.750, P<0.001). Multivariate binary logistic regression analysis showed that gender, age, serum ß2-MG, and hemodialysis (HD) adequacy (Kt/V urea) were independent risk factors for CVC in MHD patients. ROC analysis showed that a ß2-MG value of 25 µg/L was the best cut-off point for predicting CVC in MHD patients. According to binary logistic regression analysis, the ß2-MG ≥25 µg/L group was 3.39 times more likely to develop CVC than the ß2-MG <25 µg/L group [odds ratio (OR), 3.39; 95% confidence interval (CI), 1.63-7.06; P=0.001]. The DT model determined that serum ß2-MG ≥25 µg/L and age >69 years were important determinants for predicting CVC in MHD patients. Conclusions: Serum ß2-MG in MHD patients has a positive correlation with the severity and occurrence of CVC.

Correlation between lactate dehydrogenase and QTc interval prolongation in maintenance hemodialysis patients: a cross-sectional study.

BACKGROUND: Corrected QT (QTc) interval prolongation is one of the common causes of sudden cardiac death in patients with maintenance hemodialysis (MHD) patients. However, there are few studies on QTc prolongation in MHD patients. The concentration of lactate dehydrogenase (LDH) in hemodialysis population increased, and LDH was associated with the mortality of MHD patients. This study aimed to investigate the relationship between QTc interval prolongation and LDH in MHD patients. METHODS: This is a cross-sectional observational study. Patients who underwent MHD for more than 3 months in the Second Affiliated Hospital of Nantong University from November 2012 to November 2019 with complete data were selected as the research subjects. The patients were divided into the normal QTc interval group and the QTc interval prolongation group. The general data of patients and clinical laboratory indicators were collected retrospectively from the electronic medical record system. Pearson correlation analysis and binary logistic regression were used to analyze the correlation between LDH and QTc interval prolongation; the cut-off value of LDH predicting QTc interval prolongation was calculated by receiver operating characteristic (ROC) curve. RESULTS: The LDH level in the prolonged QTc interval group was significantly higher than that in the normal group (301.96±110.91 vs. 215.39±67.65, t=-8.03, P<0.001). QTc interval and LDH (r=0.386) were positively correlated. Binary logistic regression analysis showed that LDH, serum potassium <4 mmol/L, serum phosphorus, and left ventricular end-diastolic diameter (LVDd) were independent related factors for QTc interval prolongation. The ROC curve results showed that LDH =220 U/L was the best cutoff point for predicting QTc interval prolongation in MHD patients, with a sensitivity of 81.45% and a specificity of 59.35%. Binary logistic regression analysis showed that the LDH >220 U/L group was 6.34 times more likely to have QTc interval prolongation than the LDH ≤220 U/L group (OR 6.34, 95% CI: 3.47-11.58, P<0.001). CONCLUSIONS: LDH in MHD patients is closely related to QTc interval prolongation. Serum LDH, ionic calcium, serum phosphorus and potassium may predict QTc interval prolongation. Monitoring related indicators can remind clinicians to intervene as soon as possible to reduce the potential risk of arrhythmia and sudden cardiac death (SCD).

Development and Validation of a Risk Nomogram Model for Predicting Community-Acquired Pressure Injury Among the Older Adults in China: A Case-Control Study.

Purpose: A predictive model of community-acquired pressure injury (CAPI) was established and validated to allow the early identification of the risk of pressure injuries by family caregivers and community workers. Patients and Methods: The participants were hospitalized patients 65 years and older from two branches of a tertiary hospital in China, one for model training set and the other for validation set. This study was a case-control study based on hospital electronic medical records. According to the presence of pressure injury at admission, patients were divided into a case group and a control group. In the model training set, LASSO regression was used to select the best predictors, and then logistic regression was used to construct a nomogram. The performance of the model was evaluated by drawing the receiver operating characteristic curve (ROC) and calculating the area under the curve (AUC), calibration analysis, and decision curve analysis. The model used a 10-fold crossover for internal and external validation. Results: The study included a total of 20,235 subjects, including 11,567 in the training set and 8668 in the validation set. The prevalence of CAPI in the training and validation sets was 2.5% and 1.8%, respectively. A nomogram was constructed including eight variables: age ≥ 80, malnutrition status, cerebrovascular accidents, hypoproteinemia, respiratory failure, malignant tumor, paraplegia/hemiplegia, and dementia. The AUC of the prediction model in the original model, internal validation, and external validation were 0.868 (95% CI: 0.847, 0.890), mean 0.867, and 0.840 (95% CI: 0.807,03.873), respectively. The nomogram showed acceptable calibration and clinical benefit. Conclusion: We constructed a nomogram to predict CAPI from the perspective of comorbidity that is suitable for use by non-specialists. This nomogram will help family caregivers and community workers with the early identification of PI risks.

The Tumor Plastic Surgery Technology versus Traditional Repair Technology on the Repair of Large-Area Skin Defects after Maxillofacial Tumor Resection: A Randomized Controlled Trial.

Objective: To explore the effect of tumor plastic surgery on the repair of large-area skin defects after maxillofacial tumor resection. Methods: 90 patients undergoing maxillofacial tumor resection in our hospital from March 2019 to March 2020 were selected and randomized 1 : 1 to receive either tumor plastic surgery (experimental group) or traditional repair (control group). The clinical efficacy and facial cosmetic improvement of the two groups were compared. The Patient and Observer Scar Assessment Scale (POSAS) was used to evaluate the surgical outcomes of the two groups, the Profile of Mood States (POMS) was used to evaluate the patients' psychological status, and the Generic Quality of Life Inventory-74 (GQOLI-74) was used to assess the quality of life of patients. Results: Total clinical effective rate of the experimental group was significantly higher than that of the control group (p < 0.001). A higher excellent rate of facial cosmetic improvement was observed in the experimental group versus the control group (p < 0.001). Significantly lower POSAS scores of the experimental group than the control group were observed (p < 0.001). The POMS scores of the experimental group after treatment were lower than those of the control group (p < 0.001). Tumor plastic surgery resulted in a remarkably higher GQOLI-74 score in the patients versus traditional repair (p < 0.001). Conclusion: Tumor plastic surgery is a promising alternative for patients undergoing maxillofacial tumor resection. It can effectively promote the recovery of facial morphology and physiological function of patients, with high clinical efficacy, so it merits promotion and application.

Zinc-finger protein 750 mitigates malignant biological behavior of oral CSC-like cells enriched from parental CAL-27 cells.

Oral squamous cell carcinoma (OSCC) is the most commonly occurring oral malignancy. Cancer stem cells (CSCs) are known to be responsible for cancer recurrence and metastasis. Zinc-finger protein 750 (ZNF750) has been reported to inhibit OSCC cell proliferation and invasion. The present study aimed to elucidate the role of ZNF750 in the inhibition of the renewal ability of CSCs derived from the OSCC cell line, CAL-27. The effects of ZNF750 on CSC-like properties were examined using aldehyde dehydrogenase (ALDH), tumor sphere formation and colony formation assays. Reverse transcription-quantitative PCR and western blotting were performed to detect the expression levels of octamer-binding transcription factor 4, sex-determining region Y-box 2, the enhancer of zeste homolog 2 (Ezh2), embryonic ectoderm development (EED) and SUZ12 polycomb repressive complex 2 subunit (SUZ12), and for the identification of genes associated with metastasis. ZNF750 effectively attenuated CSC-like cell self-renewal abilities; ZNF750 decreased the ALDH-positive cell population, tumor sphere and colony formation abilities, cell viability and stemness factors. Furthermore, the expression levels of Ezh2, EED and SUZ12 were decreased by ZNF750. ZNF750 inhibited MMP1, 3, 9 and 13 expression levels, and decreased the cell invasion and migratory abilities. Moreover, the expression of tissue inhibitors of matrix metalloproteinases-1 was increased by ZNF750. However, opposite effects were observed following the knockdown of the ZNF750 gene. Overall, the present study demonstrated that ZNF750 has the potential to inhibit the renewal of CSC-like cells enriched from parental CAL-27 cells.

ZNF750 exerted its Antitumor Action in Oral Squamous Cell Carcinoma by regulating E2F2.

Cell cycle activator E2F transcription factor 2 (E2F2) play a key role in tumor development and metastasis. Previous RNA sequence analysis (GSE134835) revealed E2F2 was significantly reduced by Zinc-finger protein 750 (ZNF750) in oral squamous cell carcinoma (OSCC). This study was aimed to determine the involvement of E2F2 in antitumor action of ZNF750. The nude mouse xenograft model was established by subcutaneously injection of stable cell line CAL-27oeZNF750 or CAL-27shZNF750. Xenograft tumor volume and tumor weight was measured. The expression of E2F2, transcriptional repressors such as enhancer of zeste 2 (Ezh2), PHD finger protein 19 (PHF19), and the genes related to cell proliferation or metastasis was studied in vivo or in vitro. Luciferase assay was performed to investigate regulation effect of ZNF750 on E2F2 luciferase activity. The involvement of E2F2 in the antitumor action of ZNF750 was studied by cotransduced ZNF750 with E2F2 lentivirus. The tumor growth and metastasis was repressed by ZNF750 manifested by reduced tumor size, tumor weight and the genes related to cell proliferation and metastasis. However, all of these were reversed by knockdown of the ZNF750 gene. Furthermore, E2F2 luciferase activity was inhibited by ZNF750. E2F2 partly blocked the antitumor action of ZNF750 manifested by increased self-renewal, invasion, migration, elevated Ezh2 and MMP13 protein expression in ZNF750 + E2F2 groups. However, silenced E2F2 further enhanced the antitumor action of ZNF750. ZNF750 depressed E2F2 activity and played a critical role in regulating transcriptional repressors for inhibiting the OSCC growth and metastasis in OSCC.

Response of microalgae Chlorella vulgaris to Cr stress and continuous Cr removal in a membrane photobioreactor.

This study evaluated the toxicity of Cr(VI) to microalgae Chlorella vulgaris, and its removal by continuous microalgae cultivation in membrane photobioreactor (MPBR). Batch cultivation in photobioreactors showed that low concentration of Cr(VI) (0.5 and 1.0 mg L-1) stimulated the growth of C. vulgaris, while 2.0 and 5.0 mg L-1 Cr(VI) in the wastewater significantly inhibited the growth of C. vulgaris. Superoxide dismutase and catalase activities that represented cellular antioxidant capacity significantly increased at 0.5 and 1.0 mg L-1 Cr(VI), and then gradually decreased with the continuous increase of Cr(VI) concentration. The content of malondialdehyde, which represents the degree of cellular oxidative damage, increased with the increase of Cr(VI) concentration and reached the peak value at 2.0 mg L-1 Cr(VI). C. vulgaris was then cultured in MPBR equipped with hollow-fiber ultrafiltration membrane module to achieve continuous removal of Cr from wastewater. With the in-situ solid-liquid separation function of the membrane module, solid retention time (SRT) and hydraulic retention time (HRT) of the reactor could be controlled separately. Experimental results showed that both SRT and HRT had significant effects on the algal biomass production and pollutants removal. During the continuous operation, MPBR achieved a maximum total Cr reduction of 50.0% at HRT of 3-day and SRT of 40-day, and a maximum volumetric removal rate of total Cr of 0.21 mg L-1 d-1 at HRT of 2-day and SRT of 40-day.

Simultaneous removal of nutrient and sulfonamides from marine aquaculture wastewater by concentrated and attached cultivation of Chlorella vulgaris in an algal biofilm membrane photobioreactor (BF-MPBR).

Microalgae based wastewater treatment has attracted increasing attention for its many advantages in recent years. In this study, a novel microalgae biofilm membrane photobioreactor (BF-MPBR) was developed for the efficient microalgae cultivation and the removal of nutrient and sulfonamides (SAs) from marine aquaculture wastewater. Two BF-MPBRs with hydraulic retention time (HRT) of 1 day and 2 days respectively were continuously operated for 70 days without harvesting microalgae. Concentrated and attached culture of marine Chlorella vulgaris was achieved in these continuous flow BF-MPBRs due to the suspended solid carriers and microfiltration membrane module in the reactors. The algal biomass productivity achieved in BF-MPBRs with HRT of 1 day and 2 days were 14.02 and 22.03 mg L-1 day-1, respectively. In addition, at the end of the cultivation, 60.4% and 45.0% of microalgae were fixed into algal biofilm in BF-MPBRs with 1 day and 2 day HRT, respectively. Compared with batch cultivation, more efficient nutrient and SAs removal performance was achieved in BF-MPBRs, although the HRT of the BF-MPBRs used in this study was only 1 or 2 days. During the stable operation stage of the BF-MPBRs, the reduction in dissolved inorganic nitrogen (DIN), dissolved inorganic phosphorus (DIP), sulfadiazine (SDZ), sulfamethazine (SMZ) and sulfamethoxazole (SMX) were found in the range of 91.0-99.6%, 92.1-98.4%, 61.0-79.2%, 50.0-76.7% and 60.8-82.1%, respectively. Therefore, nutrient and SAs were simultaneously and efficiently removed from marine aquaculture wastewater by microalgae cultivation in BF-MPBR.

EZH2 inhibition promotes ANGPTL4/CREB1 to suppress the progression of ulcerative colitis.

AIMS: Enhancer of zeste homolog 2 (EZH2) is associated with ulcerative colitis development. However, the mechanism of EZH2 in ulcerative colitis progression remains unclear. MAIN METHODS: Lipopolysaccharide (LPS)-treated Caco-2 cells and dextran sodium sulfate (DSS)-treated mice were used as model of ulcerative colitis. The levels of EZH2, angiopoietin-like 4 (ANGPTL4) and cyclic adenosine monophosphate response element-binding protein 1 (CREB1) were tested via quantitative real-time polymerase chain reaction (qRT-PCR) and western blot. Cell viability and apoptosis was measured via 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl-tetrazolium bromide or flow cytometry. The abundances of inflammatory cytokines were examined via qRT-PCR and enzyme-linked immunosorbent assay. The association between EZH2 and ANGPTL4 was explored via chromatin immunoprecipitation. The colon damage in DSS-treated mice was investigated by colon length, histological analysis, inflammatory response and apoptosis. KEY FINDINGS: LPS induced viability inhibition, inflammatory response and apoptosis in Caco-2 cells. EZH2 expression was increased but ANGPTL4 and CREB1 levels were decreased in LPS-challenged Caco-2 cells. Overexpression of ANGPTL4 or CREB1 suppressed LPS-induced damage in Caco-2 cells. EZH2 could target ANGPTL4 to mediate CREB1 expression. Inhibition of EZH2 suppressed LPS-caused injury. Moreover, knockdown of ANNGPTL4 or CREB1 attenuated the role of EZH2 inhibition. DSS caused the reduced colon length and increased inflammatory response as well as apoptosis. EZH2 expression was up-regulated but ANGPTL4 and CREB1 expression were down-regulated in DSS-treated mice. SIGNIFICANCE: Inhibition of EZH2 declined LPS-induced injury in Caco-2 cells by mediating ANGPTL4 and CREB1, indicating the potential of EZH2 in treatment of ulcerative colitis.

rBMSC/Cav-1F92A Mediates Oxidative Stress in PAH Rat by Regulating SelW/14-3-3η and CA1/Kininogen Signal Transduction.

BACKGROUND/OBJECTIVES: Carbonic anhydrase 1 (CA1)/kininogen and selenoprotein W (SelW)/14-3-3η signal transduction orchestrate oxidative stress, which can also be regulated by nitric oxide (NO). The mutated caveolin-1 (Cav-1F92A) gene may enhance NO production. This study explored the effect of Cav-1F92A-modified rat bone marrow mesenchymal stem cells (rBMSC/Cav-1F92A) on oxidative stress regulation through CA1/kininogen and SelW/14-3-3η signal transduction in a rat model of monocrotaline- (MCT-) induced pulmonary arterial hypertension (PAH). METHOD: PAH was induced in rats through the subcutaneous injection of MCT. Next, rBMSC/Vector (negative control), rBMSC/Cav-1, rBMSC/Cav-1F92A, or rBMSC/Cav-1F92A+L-NAME were administered to the rats. Changes in pulmonary hemodynamic and vascular morphometry and oxidative stress levels were evaluated. CA1/kininogen and SelW/14-3-3η signal transduction, endothelial nitric oxide synthase (eNOS) dimerization, and eNOS/NO/sGC/cGMP pathway changes were determined through real-time polymerase chain reaction, Western blot, or immunohistochemical analyses. RESULTS: In MCT-induced PAH rats, rBMSC/Cav-1F92A treatment reduced right ventricular systolic pressure, vascular stenosis, and oxidative stress; downregulated CA1/kininogen signal transduction; upregulated SelW/14-3-3η signal transduction; and reactivated the NO pathway. CONCLUSIONS: In a rat model of MCT-induced PAH, rBMSC/Cav-1F92A reduced oxidative stress by regulating CA1/kininogen and SelW/14-3-3η signal transduction.

A mathematical model of chromosome recombination-induced drug resistance in cancer therapy.

Cytotoxic chemotherapeutics are common treatment methods of many cancers, and patients are often dosed at maximum tolerated dose (MTD), which is trying to eliminate cancer cells as much as possible. However, highly doses chemotherapy may induce unexpected gene mutations or DNA recombinations, which in turn result in unpredictable outcomes and drug resistance. In this study, we focus on the occurrence of DNA recombinations, and present a mathematical model for the influence of genomic disorder due to chemotherapy, and investigate how it may lead to drug resistance. We show that there is an optimal dose so that the tumor cells number is minimum at the steady state, which suggests the existence of an optimal dose of chemotherapy below the MTD. Model simulations show that when the dose is either low or high, the tumor cancer cells number may maintain a higher level steady state, or even sustained oscillations when the dose is too high, which are clinically inappropriate. Our results provide a theoretical study on the dose control of chemotherapy in cancer therapy.

Genetic factors for idiopathic choroidal neovascularization.

Objective: The aim of this study was to investigate genetic factors associated with idiopathic choroidal neovascularization (ICNV). Methods: We conducted a case-control study including 69 cases with ICNV and 114 controls who underwent cataract surgery. Single nucleotide polymorphisms (SNPs) from genes reported to be related to AMD, CNV and uveitis were selected for this study. Results: In an univariate analysis, the rs669676 SNP located in the COL8A1 gene was associated with the proportion of people who has idiopathic CNV ( X2 = 9.3453, corrected p-value = 0.1). For the rs669676 SNP, minor allele homozygotes, in the dominant model of genotype analysis (GG versus AA-GA), it showed significant differences in the ICNV group vs controls (p = .01, OR = 1.219 (95%CI: 1.04-1.429)). Conclusions: The rs669676 SNP located in the COL8A1 gene may contribute to a genetic susceptibility for ICNV.

Effect of organic carbon to nitrogen ratio in wastewater on growth, nutrient uptake and lipid accumulation of a mixotrophic microalgae Chlorella sp.

This study investigated the biomass/lipid production, nutrient removal and fatty acid composition of an isolated mixotrophic microalga (Chlorella sp. G-9) cultured in simulated wastewater with different TOC/TN ratio. As the TOC/TN ratio of wastewater increased from 0 to 24, the growth rate of Chlorella sp. G-9 increased gradually, but did not increase further at 30. Nutrient removal was related to microalgae growth. In the wastewater with TOC/TN ratio of 24 and 30, 99.58% and 99.61% nitrogen was removed, respectively. In conditions of initial TOC/TN ratios of 24 and 30, Chlorella sp. G-9 could accumulate lipid as high as 35.3% and 36.5%, respectively. The corresponding lipid productivities were 34.2 and 32.6 mg L-1 d-1, respectively, which were 13.7 and 13.0 times higher than those in photoautotrophic condition. Increasing the initial TOC/TN ratio of the wastewater could slightly increase the saturated degree in fatty acid, thereby improving the stability of biodiesel.

EPCR promotes MGC803 human gastric cancer cell tumor angiogenesis in vitro through activating ERK1/2 and AKT in a PAR1-dependent manner.

The endothelial cell protein C receptor (EPCR) serves a key role in activated protein C (APC)-mediated cytoprotective effects in endothelial cells, and is involved in the development of certain types of human cancer. To the best of our knowledge, the present study is the first to demonstrate that EPCR may exert effects on gastric cancer angiogenesis in vitro. To detect microvessel density (MVD), the microvascular endothelial cells were stained for cluster of differentiation (CD)31 and CD34 in 61 cases of surgical resection of gastric carcinoma tissues, and the association between the expression of EPCR protein and MVD was analyzed. In addition, to analyze the effect of EPCR expressed by gastric cancer cells on the proliferation, migration and angiogenic abilities of endothelial cells, human umbilical vein endothelial cells (HUVECs) were cultured with tumor-conditioned medium derived from EPCR knockdown or protease-activated receptor 1 (PAR1)-blocked MGC803 gastric cancer cells. A CCK-8 assay was used to assess the proliferation ability of the HUVECs. A Transwell assay was performed to assess the migration ability of the HUVECs and a Matrigel-based tube formation assay was used to assess the angiogenic activity of the HUVECs. The results demonstrated that the expression of EPCR was correlated with the MVD of gastric cancer tissues. When cultured with tumor-conditioned medium derived from EPCR knockdown or PAR1-blocked MGC803 cells, the proliferation, migration and tubules formation abilities of HUVECs were markedly inhibited markedly. The expression of phosphorylated (p)-extracellular signal regulated kinase 1/2, p-protein kinase B (AKT; s473) and p-AKT (T308) in the HUVECs was decreased. In addition, EPCR knockdown inhibited PAR1 activation in the MGC803 cells. These results indicated that the expression of EPCR in gastric cancer cell line MGC803 contributes to tumor angiogenesis in vitro by activating ERK1/2 and AKT, and that this effect of EPCR is dependent on PAR1 activation.

Integrin alpha5beta1 suppresses rBMSCs anoikis and promotes nitric oxide production.

BACKGROUND: Cell based therapy has been heralded as a novel, promising therapeutic strategy for cardiovascular diseases including pulmonary arterial hypertension (PAH). However, the low survival rate after transplantation due to cell death via anoikis is a major obstacle in stem cell therapy. Cells adhesion via Integrin alpha5beta1 (ITGA5B1) has a tendency to exert higher maximum forces. The present study aimed to evaluate the potential protective effect of ITGA5B1 on rat bone marrow mesenchymal stem cells (rBMSCs) from anoikis. METHODS: Mononuclear cells were isolated by density gradient centrifugation with Ficoll, and rBMSCs cell surface markers were evaluated by flow cytometry. Osteogenic and adipocyte differentiation was determined by Alizarin Red S and Oil Red O staining respectively. The expression of Integrin A5 (ITGA5), Integrin B1 (ITGB1), eNOS and actived-caspase-3 mRNA or protein was confirmed by qPCR and western-blot. Cell adhesion, cell viability, anoikis and the migration of rBMSCs were also evaluated. Nitric oxide (NO) production was detected by the greiss assay. RESULTS: Co-infected with Integrin A5 and B1 lentivirus to rBMSCs increased ITGA5 and ITGB1 mRNA and protein expression. ITGA5B1 enhanced the cell adhesion, cell viability, cell migration and NO production but reduced the cell anoikis in rBMSCs/ITGA5B1 groups. CONCLUSION: Transduction of rat rBMSCs with ITGA5B1 lentivirus could prevent cell anoikis and increase NO production.

Immunoregulatory Role of MicroRNA-21 in Macrophages in Response to Bacillus Calmette-Guerin Infection Involves Modulation of the TLR4/MyD88 Signaling Pathway.

BACKGROUND/AIMS: The purpose of this study is to explore the immunoregulatory role of microRNA-21 (miR-21) targeting of the TLR4/MyD88 signaling pathway in macrophages in response to Bacillus Calmette-Guerin (BCG) infection. METHODS: After infection with BCG, mouse RAW246.7 cells were assigned into control, BCG, miR-21 mimic + BCG, mimic-negative control (NC) + BCG, miR-21 inhibitor + BCG, inhibitor-NC + BCG, BCG + TAK242 (an inhibitor of the TLR4 signaling pathway), and miR-21 inhibitor + TAK242 + BCG groups. Western blotting and qRT-PCR were used to detect the expression of miR-21, TLR4 and MyD88. The levels of TNF-a, IL-6 and IL-10 were detected by enzyme-linked immunosorbent assay (ELISA). Cell viability was measured using an MTT assay. Cell apoptosis and necrosis rates were detected using flow cytometry. RESULTS: Compared with the control group, miR-21 expression and levels of TNF-a, IL-6 and IL-10, as well as cell apoptosis and necrosis rates, were elevated, while expression of TLR4 and MyD88, as well as cell viability, were reduced in BCG infection groups. Compared with the BCG group, miR-21 expression was increased in the miR-21 mimic + BCG group but decreased in the miR-21 inhibitor + BCG and miR-21 inhibitor + TAK242 + BCG groups. The expression of TLR4 and MyD88, as well as the cell viability, were decreased, while levels of TNF-a, IL-6 and IL-10, as well as cell apoptosis and necrosis rates, were increased in the miR-21 mimic + BCG and TAK242 + BCG groups. The opposite trends were found in the miR-21 inhibitor + BCG group. Compared with the TAK242 + BCG group, the miR-21 inhibitor + TAK242 + BCG group had higher expression of TLR4 and MyD88 as well as higher cell viability and lower levels of TNF-a, IL-6, IL-10, cell apoptosis and necrosis rates. However, the miR-21 inhibitor + TAK242 + BCG group exhibited the opposite trends when compared with the miR-21 inhibitor + BCG group. CONCLUSION: Our results suggest that miR-21 can negatively modulate the TLR4/MyD88 signaling pathway, resulting in decreased cell viability, increased cell apoptosis and increased levels of inflammatory factors following BCG infection in macrophages.

Knockdown of SKA1 gene inhibits cell proliferation and metastasis in human adenoid cystic carcinoma.

The spindle and kinetochore-associated complex subunit 1(SKA1) is a newly discovered gene, which has been associated with mitosis and tumorigenesis. However, its role insalivary adenoid cystic carcinoma (SACC) is still unknown, and the invasive and metastatic mechanism in SACC is still unclear. To explore the molecular mechanism of SKA1 in the process of malignant proliferation and metastasis in adenoid cystic cancer (ACC) cells, we employed lentivirus-mediated short hairpin RNA to knockdown SKA1 in SACC-83 cells. The results demonstrated that the lentivirus-mediated shRNA-targeting SKA1 lead to a significant down-regulation of SKA1 expression. Knockdown of SKA1 inhibited cell proliferation, cell invasion, migration and the cell cycle arrest. Furthermore, knockdown of SKA1 reduced the Ndc80, CDK4, Cyclin D1, Cyclin E1, Cyclin B1 and matrix metalloproteinase-9 (MMP-9) protein expression, but increased the p27 protein expression. These findings indicated that SKA1 might be a promising target for cancer gene therapy in human ACC.

[Clinical evaluation of the prevalence of endodontic interappointment emergencies during intracanal medication before and after root canal preparation].

PURPOSE: To evaluate the incidence of pain during intracanal medication before and after root canal instrumentation for therapeutic reference. METHODS: Three hundred and twenty four teeth were selected in this study, and the teeth received root canal therapy with crown-down technique using ProTaper rotary system, followed by calcium hydroxide intracanal medication. The degree of reported pain was recorded as 4 levels after treatment according to the Negm criteria. The incidence of endodontic interappointment emergencies (EIAE) at various recording times starting from root canal preparation was evaluated based on the results analyzed with MATLAB and SPSS19.0 software package. RESULTS: All patients reported of pain immediately after root canal therapy and calcium hydroxide intracanal medication, of which 1.54% were diagnosed as EIAE, the prevalence and degree of pain were increased. The prevalence of EIAE reached 25.62% one day after treatment, and decreased to 3.70% 3 days later. Most of the pain released without treatment; however, few patients experienced severe pain. The relationship between the EIAE percentage and time duration after treatment was fitted by using MATLAB as P(t)=100%×0.066t exp[-0.076t] (t: hour; RMSE=3.91). The patient started to report free of pain 6 hours after the treatment, 33.64% of the patients were painless 1 week after treatment, and 82.71% were painless 2 weeks later. There was significant difference between the pain level before and 2 weeks after treatment (P<0.01). CONCLUSIONS: During root canal preparation and intracanal medication, the prevalence and degree of pain were increased immediately after treatment, decreased 3 days later, and most of the pain was released 2 weeks after treatment.