Bioaccumulation and Trophodynamics of Novel Brominated Flame Retardants (NBFRs) in Marine Food Webs from the Arctic and Antarctic Regions.

The pervasive contamination of novel brominated flame retardants (NBFRs) in remote polar ecosystems has attracted great attention in recent research. However, understanding regarding the trophic transfer behavior of NBFRs in the Arctic and Antarctic marine food webs is limited. In this study, we examined the occurrence and trophodynamics of NBFRs in polar benthic marine sediment and food webs collected from areas around the Chinese Arctic Yellow River Station (n = 57) and Antarctic Great Wall Station (n = 94). ∑7NBFR concentrations were in the range of 1.27-7.47 ng/g lipid weight (lw) and 0.09-1.56 ng/g lw in the Arctic and Antarctic marine biota, respectively, among which decabromodiphenyl ethane (DBDPE) was the predominant compound in all sample types. The biota-sediment bioaccumulation factors (g total organic carbon/g lipid) of NBFRs in the Arctic (0.85-3.40) were 4-fold higher than those in the Antarctica (0.13-0.61). Trophic magnification factors (TMFs) and their 95% confidence interval (95% CI) of individual NBFRs ranged from 0.43 (95% CI: 0.32, 0.60) to 1.32 (0.92, 1.89) and from 0.34 (0.24, 0.49) to 0.92 (0.56, 1.51) in the Arctic and Antarctic marine food webs, respectively. The TMFs of most congeners were significantly lower than 1, indicating a trophic dilution potential. This is one of the very few investigations on the trophic transfer of NBFRs in remote Arctic and Antarctic marine ecosystems, which provides a basis for exploring the ecological risks of NBFRs in polar regions.

Occurrence and Trophic Transfer of Polychlorinated Naphthalenes (PCNs) in the Arctic and Antarctic Benthic Marine Food Webs.

Information about the occurrence and trophic transfer of polychlorinated naphthalenes (PCNs) in polar ecosystems is vital but scarce. In this study, PCNs were analyzed in benthic marine sediment and several biological species, collected around the Chinese polar scientific research stations in Svalbard in the Arctic and South Shetland Island in Antarctica. Total PCNs in biota ranged from 28 to 249 pg/g of lipid weight (lw) and from 11 to 284 pg/g lw in the Arctic and Antarctic regions, respectively. The concentrations and toxic equivalent (TEQ) of PCNs in polar marine matrices remained relatively low, and the compositions were dominated by lower chlorinated homologues (mono- to trichlorinated naphthalenes). Trophic magnification factors (TMFs) were calculated for congeners, homologues, and total PCNs in the polar benthic marine food webs. Opposite PCN transfer patterns were observed in the Arctic and Antarctic regions, i.e., trophic dilution and trophic magnification, respectively. This is the first comprehensive study of PCN trophic transfer behaviors in remote Arctic and Antarctic marine regions, providing support for further investigations of the biological trophodynamics and ecological risks of PCNs.

Evolution between bright and dark pulses in a MoxW1-xTe2 based fiber laser.

We proposed an erbium-doped fiber laser mode-locked with a MoxW1-xTe2-based nonlinear optical modulator for the first time to our best knowledge. This fiber laser can deliver bright pulses, bright-dark pulse pairs, dark pulses, bright-dark-bright pulses, and dark-dark-bright pulses. The modulation depth and saturation intensity of the MoxW1-xTe2-based saturable absorber were about 7.8% and 8.6 MW/cm2, respectively. When 10% of the laser in the cavity was output, conventional soliton pulses with central wavelength of 1560.1 nm can be obtained in the cavity. When 70% of the laser was output, dual-wavelength domain-wall dark pulses appeared in the laser cavity. This experiment revealed that an appropriate increase in the ratio of output energy can improve the chance of dark pulses in fiber lasers. The mode-locking states in this fiber laser can evolve with each other between bright pulses, bright-dark pulse pairs and dark pulses by adjusting the polarization controller. The results indicated that the MoxW1-xTe2 can be used to make modulators for generating dark pulses. Furthermore, our work will be of great help to improve the chance of the generation of dark pulse in fiber lasers.

High-Energy Mode-Locked Pulse Er-Doped Fiber Laser-Based GeTe as Saturable Absorber.

High-energy Er-doped fiber laser with high conversion efficiency is reported, which is mode-locked by a germanium telluride (GeTe)-based saturable absorber (SA). By adjusting the direction of the polarization controller (PC), a high-energy pulse with a central wavelength of 1533.1 nm and a fundamental repetition frequency of 1.58 MHz is achieved. Under the pump power of 450.1 mW, the maximum average output power is 50.48 mW, and the single-pulse energy is 32 nJ. It is worth noting that the optical-to-optical conversion efficiency has reached about 11.2%. The experimental results indicate that GeTe performs excellently as SAs for obtaining mode-locked fiber lasers and plays an extremely important role in high-energy fiber lasers.

Effect and mechanism of chlorogenic acid on cognitive dysfunction in mice by lipopolysaccharide-induced neuroinflammation.

Background: Neuroinflammation is an important factor causing numerous neurodegenerative pathologies. Inflammation can lead to abnormal neuronal structure and function and even death, followed by cognitive dysfunction. There is growing evidence that chlorogenic acid has anti-inflammatory effects and immunomodulatory activity. Purpose: The aim of this study was to elucidate the potential targets and molecular mechanisms of chlorogenic acid in the treatment of neuroinflammation. Methods: We used the lipopolysaccharide-induced neuroinflammation mouse model and the lipopolysaccharide-stimulated BV-2 cells in vitro model. Behavioral scores and experiments were used to assess cognitive dysfunction in mice. HE staining and immunohistochemistry were used to assess neuronal damage in the mouse brain. Immunofluorescence detected microglia polarization in mouse brain. Western blot and flow cytometry detected the polarization of BV-2 cells. The migration of BV-2 cells was detected by wound healing assay and transwell assay. Potential targets for chlorogenic acid to exert protective effects were predicted by network pharmacology. These targets were then validated using molecular docking and experiments. Results: The results of in vivo experiments showed that chlorogenic acid had an obvious ameliorating effect on neuroinflammation-induced cognitive dysfunction. We found that chlorogenic acid was able to inhibit BV-2 cells M1 polarization and promote BV-2 cells M2 polarization in vitro while also inhibiting the abnormal migration of BV-2 cells. Based on the network pharmacology results, we identified the TNF signaling pathway as a key signaling pathway in which chlorogenic acid exerts anti-neuroinflammatory effects. Among them, Akt1, TNF, MMP9, PTGS2, MAPK1, MAPK14, and RELA are the core targets for chlorogenic acid to function. Conclusion: Chlorogenic acid can inhibit microglial polarization toward the M1 phenotype and improve neuroinflammation-induced cognitive dysfunction in mice by modulating these key targets in the TNF signaling pathway.

Legacy and novel brominated flame retardants in air of Ny-Ålesund, Arctic from 2011 to 2019.

Concentrations of polybrominated diphenyl ethers (PBDEs) and novel brominated flame retardants (NBFRs) in the atmosphere of Ny-Ålesund, Svalbard, were investigated. Passive air samples were collected for eight consecutive one-year periods from August 2011 to August 2019 at seven Arctic sampling sites. High-resolution gas chromatography coupled with high-resolution mass spectrometry (HRGC-HRMS) and gas chromatography coupled with election capture negative ionization mass spectrometry (GC-NCI-MS) were employed for PBDE and NBFR analysis, respectively. The median concentrations of Æ©11PBDEs and Æ©6NBFRs were 0.6 pg/m3 and 4.0 pg/m3, respectively. Hexabromobenzene and BDE-47 were the most abundant NBFR and PBDE congeners in the atmosphere, accounting for 31% and 24% of Æ©NBFR and Æ©PBDE concentrations, respectively. Æ©NBFR concentration was approximately six times higher than that of Æ©PBDEs in the same samples. Among NBFRs, the concentrations of 1,2,3,4,5-pentabromobenzene, 2,3,4,5,6-pentabromobenzene, and 2,3-dibromopropyl-2,4,6-tribromophenyl ether showed increasing temporal variations, with estimated doubling times of 3.0, 3.3, and 2.8 years, respectively. The concentrations of almost all PBDE congeners showed a decreasing variation, with halving times ranging from 2.1 to 9.5 years.

Novel brominated and organophosphate flame retardants in the atmosphere of Fildes Peninsula, West Antarctica: Continuous observations from 2011 to 2020.

Novel brominated flame retardants (NBFRs) and organophosphate esters (OPEs) have been widely detected in various environmental matrices worldwide and raised public concerns in recent years. However, few studies reported their occurrence and temporal trend in Antarctic air. In this study, concentrations, distribution, and temporal trends of NBFRs and OPEs in the air of Fildes Peninsula, West Antarctica, were investigated using XAD resin-based passive air sampling from January 2011 to January 2020. Air concentrations of the total OPEs (Σ7OPEs) were one to two orders of magnitude higher than those of the total NBFRs (Σ6NBFRs). Decabromodiphenyl ethane and tris(2-chloroethyl) phosphate were the most abundant NBFR and OPE congeners, respectively. Significant positive correlations were observed among hexabromobenzene, pentabromoethylbenzene, and pentabromotoluene, indicating that their occurrence in Antarctic air may be affected by similar sources. No spatial differences in any of the NBFR and OPE congeners were observed, implying minor impact from local scientific research stations. Linear regression analysis was used to evaluate the temporal trends of NBFRs and OPEs in Antarctic air, with decreasing trends observed for Σ6NBFRs and Σ7OPEs. This is one of the rare studies providing a comprehensive investigation of the temporal trends in NBFRs and OPEs in Antarctic air and highlights concern regarding the contamination of these chemicals in remote polar regions.

Polychlorinated naphthalenes (PCNs) in soils and plants from Svalbard, Arctic: Levels, distribution, and potential sources.

As persistent organic pollutants (POPs) newly banned by the Stockholm Convention, polychlorinated naphthalenes (PCNs) have been widely detected in various environmental matrices. To date, however, the occurrence of PCNs in soils and plants in the Arctic environment has not been reported. In the current study, the concentrations and distribution of PCNs in Arctic soils and plants from Svalbard were analyzed. Total PCN concentrations ranged from 5.3 to 2550 pg/g dry weight (dw) in soils and 77 to 870 pg/g dw in plants. The higher levels of PCNs near the research stations and Longyearbyen town highlighted the significant influence of local anthropogenic emission sources. The composition of PCNs in Arctic soils and plants was dominated by lower chlorinated homologues, especially mono- to trichlorinated naphthalenes, which accounted for over 80 % of total PCNs in the soil and plant samples. The correlation analysis indicated the potential influences of total organic carbon (TOC) content on PCN concentrations in the soil, and octanol-air partition coefficients (KOA) or octanol-water partition coefficients (KOW) on PCN accumulation from soils to plants. To the best of our knowledge, this is the first study to report on the concentration and distribution of PCNs in Arctic soils and plants.

Organophosphate esters in Arctic air from 2011 to 2019: Concentrations, temporal trends, and potential sources.

Concentrations of seven organophosphate ethers (OPEs) were quantified in passive air samples deployed for eight consecutive one-year periods from August 2011 to August 2019 at seven sampling sites in the area of Ny-Ålesund, Svalbard, Arctic. Non-chlorinated and chlorinated OPEs were approximately equally abundant and the mean atmospheric concentration for the sum of OPEs was around 300 pg/m3. Levels of OPEs were two orders of magnitude higher than those of polybrominated diphenyl ethers in the sampling regions, likely a result of efficient long-range transport and higher environmental release rates. For the two most abundant compounds, tris(2-chloroethyl) phosphate and tris-n-butyl phosphate, increasing temporal trends in atmospheric concentrations were observed, with estimated doubling times of 2.9 and 4.2 years, respectively. Slightly elevated OPE levels at two sampling sites in the vicinity of a research station and the local airport suggest the possible influence of local contamination sources. Re-volatilization from glaciers may also influence levels of OPE in the Arctic atmosphere.

Associations between Indoor Environmental Quality and Infectious Diseases Knowledge, Beliefs and Practices of Hotel Workers in Wuhan, China.

Knowledge, beliefs, and practices regarding infectious diseases are key elements that ensure practitioners' health and safety. It is important to carry out such a survey in hotels. This study aims to determine the levels of knowledge, beliefs, and practices regarding infectious diseases among practitioners and their associations with the environmental quality of hotels in Wuhan, China. We surveyed infectious disease knowledge, beliefs, and practices of practitioners in 18 hotels and detected these hotels' environment, including physical factors of temperature, humidity, noise, and the indoor air quality of benzene, toluene, xylene, formaldehyde, CO, CO2, the total count of fungi, aerobic plate count, PM10, and PM2.5. 128 practitioners were included, and 28.9% were male. The questionnaire included knowledge, beliefs, and practices regarding infectious diseases. Our study found moderate levels of knowledge and beliefs, and good health practices. People's beliefs toward COVID-19 were correlated significantly with their knowledge (p < 0.05). Beliefs and health practices were correlated significantly with environmental quality (p < 0.05). However, the environmental quality was correlated negatively with the classification of hotels. Conclusively, despite the good health practices of practitioners, the knowledge and beliefs toward infectious diseases need to strengthen. Hotels should emphasize health education in practitioners and the improvement of environmental hygiene. Integrating all three components into a comprehensive environmental promotion program is warranted.

Accumulation and influencing factors of novel brominated flame retardants in soil and vegetation from Fildes Peninsula, Antarctica.

The concentrations and distributions of nine novel brominated flame retardants (NBFRs) were analyzed in soil, lichen (Usnea aurantiaco-atra), and moss (Sanionia uncinata) samples collected from the Chinese Antarctic Great Wall Station and surrounding Fildes Peninsula area in west Antarctica. Total NBFR concentrations ranged from 61.2-225 pg/g dry weight (dw) in soil, 283-1065 pg/g dw in moss, and 135-401 pg/g dw in lichen, respectively. Decabromodiphenyl ethane (DBDPE) was the dominant NBFR in all samples, accounting for 65.2%, 50.1%, and 72.4% of cumulative NBFR concentration in soil, moss, and lichen, respectively. The concentrations of NBFRs in plant samples were higher than those in soil, which may be related to plant bioaccumulation. Significant log/log-linear correlations (p < 0.05) were found between the concentrations of BEHTEBP and total organic carbon (TOC) in soil, and between DBDPE and lipid content in mosses, indicating that TOC and lipid content potentially affect certain NBFRs in Antarctic soil and moss. This study presents the first report on NBFR contamination in soil and various vegetation in Antarctica.

NPFF increases fusogenic proteins syncytin 1 and syncytin 2 via GCM1 in first trimester primary human cytotrophoblast cells.

Neuropeptide FF (NPFF) is well-known for its roles in the central nervous system. Despite studies demonstrating that NPFF receptor 2 (NPFFR2) mRNA is highest in placenta, nothing is known about NPFF-NPFFR2 functions in placental development. Here, we investigated the effects of NPFF-NPFFR2 on expression of syncytial [human chorionic gonadotropin (hCG) ß] and fusogenic [syncytin 1, syncytin 2, and glial cells missing 1 (GCM1)] genes in first trimester primary human cytotrophoblast cells. By analyzing two publicly available microarray data sets, we found that NPFF is consistently expressed throughout gestation whereas NPFFR2 increases in first trimester and is elevated in placenta samples from women with preeclampsia. Immunohistochemistry showed that NPFFR2, syncytin 1/2, and GCM1 each displayed unique patterns of expression among different trophoblast populations in first trimester placenta. Treatment of primary human cytotrophoblast cells with NPFF increased the mRNA and protein levels of hCG ß, syncytin 1, syncytin 2, and GCM1; and knockdown of NPFFR2 abolished these effects. Interestingly, GCM1 mediated NPFF-induced upregulation of syncytin 1 and syncytin 2, but not hCG ß, in primary human cytotrophoblasts. Our results demonstrate that NPFF acts via NPFFR2 to enhance production of hCG ß and promote GCM1-dependent expression of syncytin 1 and 2 in human cytotrophoblasts.

TGFß1 induces endometrial cancer cell adhesion and migration by up-regulating integrin αvß3 via SMAD-independent MEK-ERK1/2 signaling.

Endometrial cancer is the most common, and second most lethal, gynecological malignancy, and its rates of incidence and death are growing. This is likely attributable to increased numbers of high-risk type II endometrial cancers which account for ~30% of cases but ~75% of deaths due to their aggressive and metastatic behaviour. Histopathological and in vitro functional studies suggest that aberrant TGFß1 signaling may contribute to endometrial cancer development and the acquisition of invasive/metastatic characteristics. However, little is known about the cellular and molecular mechanisms of TGFß1 in high-risk endometrial cancers. In the present study, we examined the roles and mechanisms of TGFß1 on cell adhesion and motility in type II endometrial cancer cell lines, KLE and HEC-1B. We show that treatment with TGFß1 increases cell adhesion to vitronectin and transwell cell migration. We also demonstrate that TGFß1 treatment increases integrin ß3 and αv mRNA and protein levels via SMAD-independent MEK-ERK1/2 signaling. Importantly, siRNA depletion or antibody-mediated blocking of integrin αvß3 reversed the effects of TGFß1 on cell adhesion and migration. Our results suggest that TGFß1-MEK-ERK1/2-integrin αvß3 signaling could contribute to the invasive behaviour of high-risk endometrial cancer by promoting cell adhesion and migration.

Sprouty2 inhibits amphiregulin-induced down-regulation of E-cadherin and cell invasion in human ovarian cancer cells.

Similar to Drosophila Sprouty (SPRY), mammalian SPRY proteins inhibit the receptor tyrosine kinase-mediated activation of cellular signaling pathways. SPRY2 expression levels have been shown to be down-regulated in human ovarian cancer, and patients with low SPRY2 expression have significantly poorer survival than those with high SPRY2 expression. In addition, epidermal growth factor receptor (EGFR) is overexpressed in human ovarian cancer and is associated with more aggressive clinical behavior and a poor prognosis. Amphiregulin (AREG), the most abundant EGFR ligand in ovarian cancer, binds exclusively to EGFR and stimulates ovarian cancer cell invasion by down-regulating E-cadherin expression. However, thus far, the roles of SPRY2 in AREG-regulated E-cadherin expression and cell invasion remain unclear. In the present study, we show that treatment with AREG up-regulated SPRY2 expression by activating the EGFR-mediated ERK1/2 signaling pathway in two human ovarian cancer cell lines, SKOV3 and OVCAR5. In addition, overexpression of SPRY2 attenuated the AREG-induced down-regulation of E-cadherin by inhibiting the induction of the E-cadherin transcriptional repressor, Snail. Moreover, SPRY2 overexpression attenuated AREG-stimulated cell invasion and proliferation. This study reveals that SPRY2 acts as a tumor suppressor in human ovarian cancer and illustrates the underlying mechanisms that can be used as possible targets for the development of novel therapeutics.

Growth differentiation factor 8 induces SKOV3 ovarian cancer cell migration and E-cadherin down-regulation.

Epithelial ovarian cancer is the most lethal gynecological malignancy because most women present with late stage disseminated disease. Epithelial-mesenchymal transition (EMT) is characterized by the down-regulation of E-cadherin and up-regulation of N-cadherin, and is a crucial event in the pathogenesis of ovarian cancer. Transforming growth factor-ß (TGF-ß) is a major regulator of EMT in many normal and neoplastic cell types. Growth differentiation factor 8 (GDF8), which also activates TGF-ß-like SMAD2/3 signaling, is best known for negatively regulating muscle growth. Though recent studies suggest that GDF8 enhances placental trophoblast cell migration, little is known about the role of GDF8 in EMT and cancer metastasis. We hypothesized that GDF8 could enhance ovarian cancer cell migration by inducing EMT. Here we demonstrate for the first time that GDF8 down-regulates E-cadherin but does not alter N-cadherin in SKOV3 ovarian cancer cells. This effect is abolished by the activin receptor-like kinase (ALK)4/5/7 inhibitor SB431542 or siRNA-mediated knockdown of ALK5, whereas knockdown of ALK4 is only partially inhibitory. GDF8 treatment increases the phosphorylation of SMAD2/3 and up-regulates the E-cadherin transcriptional repressors Snail and Slug; and these effects are abolished by pre-treatment with SB431542. Knockdown of common SMAD4 fully reverses the effects of GDF8 on E-cadherin and partially attenuates its effects on Snail and Slug. Importantly, GDF8 treatment increases SKOV3 cell migration and this effect is blocked by SB431542. Our study suggests that GDF8 promotes ovarian cancer cell migration via ALK4/5-SMAD2/3-E-cadherin signaling.

TGF-ß1 stimulates migration of type II endometrial cancer cells by down-regulating PTEN via activation of SMAD and ERK1/2 signaling pathways.

PTEN acts as a tumor suppressor primarily by antagonizing the PI3K/AKT signaling pathway. PTEN is frequently mutated in human cancers; however, in type II endometrial cancers its mutation rate is very low. Overexpression of TGF-ß1 and its receptors has been reported to correlate with metastasis of human cancers and reduced survival rates. Although TGF-ß1 has been shown to regulate PTEN expression through various mechanisms, it is not yet known if the same is true in type II endometrial cancer. In the present study, we show that treatment with TGF-ß1 stimulates the migration of two type II endometrial cancer cell lines, KLE and HEC-50. In addition, TGF-ß1 treatment down-regulates both mRNA and protein levels of PTEN. Overexpression of PTEN or inhibition of PI3K abolishes TGF-ß1-stimulated cell migration. TGF-ß1 induces SMAD2/3 phosphorylation and knockdown of common SMAD4 inhibits the suppressive effects of TGF-ß1 on PTEN mRNA and protein. Interestingly, TGF-ß1 induces ERK1/2 phosphorylation and pre-treatment with a MEK inhibitor attenuates the suppression of PTEN protein, but not mRNA, by TGF-ß1. This study provides important insights into the molecular mechanisms mediating TGF-ß1-induced down-regulation of PTEN and demonstrates an important role of PTEN in the regulation of type II endometrial cancer cell migration.

Activin B promotes endometrial cancer cell migration by down-regulating E-cadherin via SMAD-independent MEK-ERK1/2-SNAIL signaling.

High-risk type II endometrial cancers account for ~30% of cases but ~75% of deaths due, in part, to their tendency to metastasize. Histopathological studies of type II endometrial cancers (non-endometrioid, mostly serous) suggest overproduction of activin B and down-regulation of E-cadherin, both of which are associated with reduced survival. Our previous studies have shown that activin B increases the migration of type II endometrial cancer cell lines. However, little is known about the relationship between activin B signaling and E-cadherin in endometrial cancer. We now demonstrate that activin B treatment significantly decreases E-cadherin expression in both a time- and concentration-dependent manner in KLE and HEC-50 cell lines. Interestingly, these effects were not inhibited by knockdown of SMAD2, SMAD3 or SMAD4. Rather, the suppressive effects of activin B on E-cadherin were mediated by MEK-ERK1/2-induced production of the transcription factor SNAIL. Importantly, activin B-induced cell migration was inhibited by forced-expression of E-cadherin or pre-treatment with the activin/TGF-ß type I receptor inhibitor SB431542 or the MEK inhibitor U0126. We have identified a novel SMAD-independent pathway linking enhanced activin B signaling to reduced E-cadherin expression and increased migration in type II endometrial cancer.

Activin B induces human endometrial cancer cell adhesion, migration and invasion by up-regulating integrin ß3 via SMAD2/3 signaling.

Endometrial cancer is the fourth most common female cancer and the most common gynecological malignancy. Although it comprises only ~10% of all endometrial cancers, the serous histological subtype accounts for ~40% of deaths due to its aggressive behavior and propensity to metastasize. Histopathological studies suggest that elevated expression of activin/inhibin ßB subunit is associated with reduced survival in non-endometrioid endometrial cancers (type II, mostly serous). However, little is known about the specific roles and mechanisms of activin B (ßB dimer) in serous endometrial cancer growth and progression. In the present study, we examined the biological functions of activin B in type II endometrial cancer cell lines, HEC-1B and KLE. Our results demonstrate that treatment with activin B increases cell migration, invasion and adhesion to vitronectin, but does not affect cell viability. Moreover, we show that activin B treatment increases integrin ß3 mRNA and protein levels via SMAD2/3-SMAD4 signaling. Importantly, siRNA knockdown studies revealed that integrin ß3 is required for basal and activin B-induced cell migration, invasion and adhesion. Our results suggest that activin B-SMAD2/3-integrin ß3 signaling could contribute to poor patient survival by promoting the invasion and/or metastasis of type II endometrial cancers.

L-carnitine and taurine synergistically inhibit the proliferation and osteoblastic differentiation of vascular smooth muscle cells.

AIM: To investigate the synergistic action of L-carnitine (LC) and taurine (TAU) on the proliferation and osteoblastic differentiation of vascular smooth muscle cells (VSMCs). METHODS: DNA and protein synthesis of VSMCs were assessed using scintillation counting. Alkaline phosphatase (ALP) activity and calcium content were determined to investigate the effects of LC and TAU on the osteoblastic differentiation and mineralization of VSMCs. TAU uptake by VSMCs was assayed. RNA interference was used to down-regulate the expression of the TAU transporter (TAUT) in rat VSMCs. RESULTS: LC and TAU synergistically inhibited the proliferation and beta-glycerophosphate (beta-GP)-induced osteoblastic differentiation of VSMCs as evidenced by the decreased [(3)H]thymidine incorporation, ALP activity and calcium deposition. Furthermore, LC stimulated the TAU uptake and TAUT expression in VSMCs. Suppression of TAUT with short hairpin RNA (shRNA) abolished the synergistic action of LC and TAU in VSMCs. CONCLUSION: The synergistic inhibitory action of LC and TAU on the proliferation and osteoblastic differentiation of VSMCs is attributable to the up-regulation of TAUT expression and TAU uptake by LC.