0D/2D mixed-dimensional perovskite constructed by thiol- and disulfide-containing ligands.

Reduced-dimensional (RD) perovskites have shown attractive chemical and physical properties for optoelectronic applications. Incorporating large organic ligands enables infinite tunability in the components and structures. Theoretically, it is feasible to apply multiple types of organic ligands in a single RD crystal to achieve multiple-dimensional perovskites. However, the coexistence of different organic ligands commonly introduces competing crystal growths that inhibit the formation of a more complex crystal structure. Herein, we report a case of mixed-dimensional (MD) perovskite single crystal containing two types of sulfide-containing ligands. We show that the application of ketones can partially oxidize organothiol ligands in the precursor solution. The resultant disulfide-based ligands can be co-incorporated with the thiol-based ligand in a single MD perovskite crystal. X-ray diffraction confirmed that the structure contains both layered and isolated inorganic components constructed by face-sharing lead halide octahedra. Unlike conventional RD structures, the MD perovskite shows an enlarged bandgap with valence band maximum and conduction band minimum being spatially separated, and isotropic optical features, as revealed by x-ray diffraction, spectroscopies, and density functional theory computation.

Unified one-electron Hamiltonian formalism of spin-orbit Jahn-Teller and pseudo-Jahn-Teller problems in tetrahedral and octahedral symmetries.

Heavy element compounds with high symmetries often feature both spin-orbit coupling and vibronic coupling. This is especially true for systems with tetrahedral and octahedral symmetries, whose electronic states may be threefold degenerate and experience complicated Jahn-Teller and pseudo-Jahn-Teller interactions. To accurately describe these interactions, high quality spin-orbit vibronic Hamiltonian operators are needed. In this study, we present a unified one-electron Hamiltonian formalism for spin-orbit vibronic interactions for systems in all tetrahedral and octahedral symmetries. The formalism covers all spin-orbit Jahn-Teller and pseudo-Jahn-Teller problems in the symmetries with arbitrary types and arbitrary numbers of vibrational modes and generates Hamiltonian expansion formulas of arbitrarily high order.

Spontaneous Polarization Effect and Photocatalytic Activity of Layered Compound of BiOIO3.

Internal polarized electric field is found to be an effective and available strategy to separate photogenerated electron-hole pairs. By this method, the efficiency of photocatalytic reactions can be obviously enhanced. Here, the layered compound of BiOIO3 with spontaneous polarization was synthesized by a simple hydrothermal method. Taking another bismuth compound BiOI as a counterpart, which has a similar layered structure, the spontaneous polarization effects of BiOIO3 were analyzed and confirmed. The photocatalytic activity of BiOIO3 and BiOI were evaluated by the degradation of methyl orange. Methyl orange was almost completely photocatalytically decomposed by BiOIO3 and BiOI in 40 and 90 min, respectively. The separation and transfer behaviors of photogenerated electron-hole pairs were investigated by a series of photoelectrochemical characterizations. It is further proved the separation and transmission efficiency of BiOIO3 are higher than those of BiOI. According to the results of density of theory calculations, the internal polarized electric field in BiOIO3 is ascribed to the spatial asymmetry of the IO3 group, which is estimated to ∼1.5 × 1010 V/m. Under the action of this internal polarized electric field, the photogenerated electrons and holes would transfer along opposite directions, i.e., photogenerated electrons and holes respectively gather at the Bi/I side and O side. Additionally, superoxide radicals (•O2-) and holes (h+) are produced during the degradation process, which are responsible for the high visible-light photocatalytic activity. Finally, the cyclic degradation test proves that its photocatalytic performance has long-term stability. Therefore, BiOIO3 polar material can be used as one of the alternative materials for efficient photocatalytic reaction.

Rapid BMI Increases and Persistent Obesity in Small-for-Gestational-Age Infants.

Purpose: In order to compensate for the early intrauterine growth restriction, small-for-gestational age (SGA) infants have "catch-up growth" after birth. Increased caloric intake has been suggested for SGA infants conventionally. It is important to determine if the early growth rate of body mass index (BMI) is associated with risk of persistent obesity later in life. In this longitudinal cohort study, we assessed the BMI of a large cohort of children who were SGA at birth to determine their risk of persistent obesity at school age (6-7 years) due to excessive weight gain in the first 3 years of life. Methods: We collected the height and weight data of 23,871 SGA babies. A polynomial function was used to fit the BMI-for-age z-score (BAZ) values of 0-6 years old SGA children and interpolate their growth trajectory. In addition, we screened out 6,959 children from 23,871 children to further evaluate the dynamic changes of early childhood BMI. We divided the school-age children into groups as non-obese (BAZ < 2) and obese (BAZ > 2), and determined the association between changes in BMI and school-age obesity. Results: From the perspective of BMI distribution, the interpolated growth trajectory indicated that SGA children reaching overweight status or developing obesity by 3 years of age, continued to have obesity until school age (R2, 0.65; R2, 0.21). The retrospective analysis showed that children who were overweight and had obesity during school age had a high BMI from early age. By analyzing the changes in early BMI, we found that the fastest growth of SGA children occurred in the early infancy before 6 months and they continued to grow rapidly for a period of time. Interestingly, former SGA children who maintained a near overweight (1 < BAZ < 2) status before the age of 2 maintained an appropriate growth rate and usually did not develop obesity. Conclusions: A rapid increase in BMI during early infancy in former SGA newborns leads to a persistent risk of obesity. The energy intake of SGA infants should appropriately meet the infants' growth needs and early BMI changes should be closely monitored for an optimal integrated management.

Redox-sensitive signaling by angiotensin II involves oxidative inactivation and blunted phosphorylation of protein tyrosine phosphatase SHP-2 in vascular smooth muscle cells from SHR.

Angiotensin II (Ang II) signaling in vascular smooth muscle cells (VSMCs) involves reactive oxygen species (ROS) through unknown mechanisms. We propose that Ang II induces phosphorylation of growth signaling kinases by redox-sensitive regulation of protein tyrosine phosphatases (PTP) in VSMCs and that augmented Ang II signaling in spontaneously hypertensive rats (SHRs) involves oxidation/inactivation and blunted phosphorylation of the PTP, SHP-2. PTP oxidation was assessed by the in-gel PTP method. SHP-2 expression and activity were evaluated by immunoblotting and by a PTP activity assay, respectively. SHP-2 and Nox1 were downregulated by siRNA. Ang II induced oxidation of multiple PTPs, including SHP-2. Basal SHP-2 content was lower in SHRs versus WKY. Ang II increased SHP-2 phosphorylation and activity with blunted responses in SHRs. Ang II-induced SHP-2 effects were inhibited by valsartan (AT(1)R blocker), apocynin (NAD(P)H oxidase inhibitor), and Nox1 siRNA. Ang II stimulation increased activation of ERK1/2, p38MAPK, and AKT, with enhanced effects in SHR. SHP-2 knockdown resulted in increased AKT phosphorylation, without effect on ERK1/2 or p38MAPK. Nox1 downregulation attenuated Ang II-mediated AKT activation in SHRs. Hence, Ang II regulates PTP/SHP-2 in VSMCs through AT(1)R and Nox1-based NAD(P)H oxidase via two mechanisms, oxidation and phosphorylation. In SHR Ang II-stimulated PTP oxidation/inactivation is enhanced, basal SHP-2 expression is reduced, and Ang II-induced PTP/SHP-2 phosphorylation is blunted. These SHP-2 actions are associated with augmented AKT signaling. We identify a novel redox-sensitive SHP-2-dependent pathway for Ang II in VSMCs. SHP-2 dysregulation by increased Nox1-derived ROS in SHR is associated with altered Ang II-AKT signaling.

[Thyroid disruption induced by pentachlorophenol].

OBJECTIVE: To assess thyroid disruption induced by sodium pentachlorophenol (PCP) using Organization for Economic Co-operation and Development (OECD) recommended TG 407 method. METHODS: A total of 30 specific pathogen free (SPF) SD adult male and female rats were randomly divided into 3 groups, and treated with water, 0.33 and 30 mg x kg(-1)x d(-1) of PCP-Na by oral gavage for consecutive 28 days, respectively. After final treatment, histological changes of thyroid were observed by hematoxylin-eosin stain, and the levels of thyroid hormones (total thyroxine (TT(4)), free thyroxine (FT(4)), total triiodothyronine (TT(3)), and free triiodothyronine (FT(3))) were determined by radioimmunoassay. The expression levels of thyroid receptors (TRalpha and TRbeta) mRNA and deiodinases (DioI, DioII and DioIII) mRNA in liver were analyzed by RT-PCR. RESULTS: In high dose group, liver weight coefficient of male and female rats were (4.82 +/- 0.42)% and (4.99 +/- 0.17)%, increased by 36.2% (t = 7.338, P < 0.01) and 41.8% (t = 8.955, P < 0.01), compared to control group ((3.54 +/- 0.14)%, (3.52 +/- 0.19)%), respectively, while the significant changes of kidney or thyroid weight were not observed. In high dose group, the levels of TT(4) and FT(4) in serum of male rats were (64.95 +/- 7.16) nmol/L and (8.16 +/- 2.29) pmol/L, and decreased by 26.6% (t = -3.999, P < 0.01) and 42.3% (t = -4.112, P < 0.01) compared to control group ((88.48 +/- 6.99) nmol/L, (14.13 +/- 1.68) pmol/L). In the same group, FT(4) in serum of female rats was (4.94 +/- 0.89) pmol/L, decreased by 55.5% (t = -3.380, P = 0.012) compared to control group ((11.10 +/- 3.40) pmol/L) and TT(3) and FT(3) in serum of female rats were (1.92 +/- 0.24) nmol/L and (3.05 +/- 0.79) pmol/L, increased by 74.5% (t = 5.263, P < 0.01) and 55.6% (t = 3.495, P < 0.01) compared to control group ((1.10 +/- 0.23) nmol/L, (1.96 +/- 0.32) pmol/L), respectively. PCP-Na didn't affect the expression levels of TRalpha, TRbeta, DioIII mRNA in high dose group, while DioII expression of male rats (0.209 +/- 0.017) down-regulated by 79.2% (t = -5.426, P < 0.01) compared to control group (1.006 +/- 0.137), and DioI expression of female rats (1.844 +/- 0.189) up-regulated by 66.6% (t = 4.359, P < 0.01) compared to control group (1.005 +/- 0.083), indicating DioI and DioII poss different sensitivity to adverse effects induced by PCP-Na between male and female rats. The histopathological results showed that PCP-Na could give rise to hyperplasia of the follicular epithelium cells, and the depletion of colloid. There were no significant changes in serum THs levels and expression of TRalpha, TRbeta, DioI-IIImRNA in low dose group. However, sporadic lymphocytic infiltration, follicles amplification in part and slightly increased in thickness of follicular cells were observed in this group. CONCLUSION: PCP is a kind of thyroid disrupting chemical.

[Effects of lycopene on cerebral ischemia-reperfusion injury in rats].

OBJECTIVE: To study the protective effects of lycopene (LP) on cerebral ischemia-reperfusion injury and oxidative stress in SD rats and the mechanism of them. METHODS: The rats were divided into five groups: normal control group, model control group, sham group and two LP groups (fed with 5 mg/kg bw or 20 mg/kg bw of lycopene daily for 15 days). The model for cerebral ischemia-reperfusion injury was established by middle cerebral artery occlusion (MCAO). The score of neurological behavior was evaluated at the 3rd and 24th hours after reperfusion. The rats were put to death 24 h after reperfusion. The size of cerebral infarction was measured. The activities of iNOS, SOD, CAT and the contents of NO and MDA in brain and serum uric acid were measured. The expressions of Bcl-2 mRNA and HIF-1alphamRNA in cortex were examined by using reverse transcription polymerase chain reaction (RT-PCR) technique. RESULTS: In comparison with the model group, the neurological deficits were milder, the volumes of cerebral infarction were smaller, the activities of SOD, CAT in brain tissue were higher, the activities of iNOS as well as the contents of NO, MDA in brain tissue and serum uric acid were lower in Lycopene groups. Compared with the model group and control group, the expression of HIF-1 alpha mRNA of cortex in the high dose lycopene (20 mg/kg bw) group was up-regulated; while the expression of Bcl-2 mRNA of cortex was up-regulated only in the low dose lycopene (5 mg/kg bw) group. CONCLUSION: There were some protective effects of oral administration of lycopene against cerebral ischemia-reperfusion injuries induced by focal cerebral ischemia and oxidative stress. The possible mechanism may be related with increasing activities of antioxidant enzymes, inhibiting lipid peroxidation, decreasing activities of iNOS,and up-regulating the expression of HIF-1alpha mRNA as well as Bcl-2 mRNA.

Regulation of the novel Mg2+ transporter transient receptor potential melastatin 7 (TRPM7) cation channel by bradykinin in vascular smooth muscle cells.

BACKGROUND: Transient receptor potential melastatin 7 (TRPM7) channels have been identified in the vasculature. However, their regulation and function remain unclear. METHODS: Here, we tested the hypothesis that bradykinin and its second messenger cAMP upregulate TRPM7, which stimulates activation of annexin-1 (TRPM7 substrate) and increases transmembrane Mg2+ transport and vascular smooth muscle cell (VSMC) migration. Human and rat VSMCs were studied. TRPM7 phosphorylation was assessed by immunoprecipitation:immunoblotting using antiphospho-serine/threonine and anti-TRPM7 antibodies. [Mg2+]i was measured by mag-fura-2. TRPM7 was downregulated by small interfering RNA and 2-aminoethoxydiphenyl borate. Annexin-1 activity was assessed by cytosol-to-membrane translocation. Cell migration and invasion, functional responses to bradykinin, were assessed in transwell chambers. RESULTS: Bradykinin increased expression of TRPM7 and annexin-1. TRPM7 was rapidly (5 min) phosphorylated on serine/threonine but not on tyrosine residues by bradykinin. [Mg2+]i was increased in bradykinin-stimulated cells (0.53 versus 0.68 mmol/l, basal versus bradykinin, P < 0.01). Annexin-1 activation was increased by bradykinin and inhibited by 2-aminoethoxydiphenyl borate. Although Hoe 140 (B2 receptor antagonist), U-73122 (phospholipase C inhibitor), 4-amino-5-(4-chlorophenyl)-7-(t-butyl)pyrazolo[3,4-d]pyrimidine (c-Src inhibitor) and chelerythrine (protein kinase C inhibitor) blocked bradykinin actions, dibutyryl-c-AMP was without effect. In small interfering RNA-transfected and in 2-aminoethoxydiphenyl borate-treated cells, bradykinin-induced Mg2+ influx and VSMC migration were reduced. CONCLUSION: Our results demonstrate that bradykinin regulates TRPM7 and its downstream target annexin-1 through phospholipase C-dependent, protein kinase C-dependent and c-Src-dependent and cAMP-independent pathways; effects are mediated through bradykinin type 2 receptor; and bradykinin regulates VSMC [Mg2+]i and migration through TRPM7. These data identify TRPM7/annexin-1/Mg2+ as a novel pathway in bradykinin signaling.

Transient receptor potential melastatin 7 ion channels regulate magnesium homeostasis in vascular smooth muscle cells: role of angiotensin II.

Magnesium modulates vascular smooth muscle cell (VSMC) function. However, molecular mechanisms regulating VSMC Mg2+ remain unknown. Using biochemical, pharmacological, and genetic tools, the role of transient receptor potential membrane melastatin 7 (TRPM7) cation channel in VSMC Mg2+ homeostasis was evaluated. Rat, mouse, and human VSMCs were studied. Reverse transcriptase polymerase chain reaction and immunoblotting demonstrated TRPM7 presence in VSMCs (membrane and cytosol). Angiotensin II (Ang II) and aldosterone increased TRPM7 expression. Gene silencing using small interfering RNA (siRNA) against TRPM7, downregulated TRPM7 (mRNA and protein). Basal [Mg2+]i, measured by mag fura-2AM, was reduced in siRNA-transfected cells (0.39+/-0.01 mmol/L) versus controls (0.54+/-0.01 mmol/L; P<0.01). Extracellular Mg2+ dose-dependently increased [Mg2+]i in control cells (Emax 0.70+/-0.02 mmol/L) and nonsilencing siRNA-transfected cells (Emax 0.71+/-0.04 mmol/L), but not in siRNA-transfected cells (Emax 0.5+/-0.01 mmol/L). The functional significance of TRPM7 was evaluated by assessing [Mg2+]i and growth responses to Ang II in TRPM7 knockdown cells. Acute Ang II stimulation decreased [Mg2+]i in control and TRPM7-deficient cells in a Na+-dependent manner. Chronic stimulation increased [Mg2+]i in control, but not in siRNA-transfected VSMCs. Ang II-induced DNA and protein synthesis, measured by 3[H]-thymidine and 3[H]-leucine incorporation, respectively, were increased in control and nonsilencing cells, but not in TRPM7 knockdown VSMCs. Our data indicate that VSMCs possess membrane-associated, Ang II-, and aldosterone-regulated TRPM7 channels, which play a role in regulating basal [Mg2+]i, transmembrane Mg2+ transport and DNA and protein synthesis. These novel findings identify TRPM7 as a functionally important regulator of Mg2+ homeostasis and growth in VSMCs.

[Mechanism of protective effects of procyanidins on liver injury induced by alcohol in mice].

OBJECTIVE: To study the mechanism of protective effects of procyanidins on liver injury model induced by alcohol in mice. METHODS: Mice were divided into 3 groups to set up the model of liver injury induced by alcohol in mice: the group of liver injury induced by alcohol, the group of procyanidins with low doses (100mg/kg), the group of procyanidins with high doses(300mg/kg). Then established normal group of control. The model of liver injury induced by alcohol in mice was used to investigate the protective effects at low and high doses procyanidins in serum indicated by alanine aminotransferase (ALT), asparate aminotransferase (AST) and in liver homogenate indicated by malonaldehyde (MDA), superoxide dismutase (SOD), reactive oxygen species (ROS). The levels of Cu,Zn-SOD mRNA in hepatic tissue and toll like receptor 4 (TLR4) mRNA were determined by using reverse transcription polymerase chain reaction (RT-PCR) technique. RESULTS: The procyanidins with low and high doses could reduce ALT, MDA, ROS content and increase SOD level, up-regulate the expression of Cu, Zn-SOD mRNA and down-regulate the expression of TLR4 mRNA. CONCLUSION: Mechanism of the protective effects on liver injuries of mice induced by alcoholmay be associated with promoting the expression of Cu, Zn-SOD and suppressing the expression of TLR4 mRNA.

Expression of a functionally active gp91phox-containing neutrophil-type NAD(P)H oxidase in smooth muscle cells from human resistance arteries: regulation by angiotensin II.

A major source of vascular smooth muscle cell (VSMC) superoxide is NAD(P)H oxidase. However, the molecular characteristics and regulation of this enzyme are unclear. We investigated whether VSMCs from human resistance arteries (HVSMCs) possess a functionally active, angiotensin II (Ang II)-regulated NAD(P)H oxidase that contains neutrophil oxidase subunits, including p22phox, gp91phox, p40phox, p47phox, and p67phox. mRNA expression of gp91phox homologues, nox1 and nox4, was also assessed in HVSMCs, human aortic smooth muscle cells, and rat VSMCs. HVSMCs were obtained from resistance arteries from gluteal biopsies of healthy subjects. gp91phox and nox4, but not nox1, were detected in HVSMCs. Nox1 and nox4, but not gp91phox, were expressed in human aortic smooth muscle cells and rat VSMCs. All NAD(P)H oxidase subunits were present in HVSMCs as detected by reverse transcriptase-polymerase chain reaction and immunoblotting. Ang II increased NAD(P)H oxidase subunit abundance. These effects were inhibited by cycloheximide. Acute Ang II stimulation (10 to 15 minutes) increased p47phox serine phosphorylation and induced p47phox and p67phox translocation. This was associated with NAD(P)H oxidase activation. In cells transfected with gp91phox antisense oligonucleotides, Ang II-mediated actions were abrogated. NADPH-induced superoxide generation was reduced by gp91ds-tat and apocynin, inhibitors of p47phox-gp91phox interactions. Our results suggest that HVSMCs possess a functionally active gp91phox-containing neutrophil-like NAD(P)H oxidase. Ang II regulates the enzyme by inducing phosphorylation of p47phox, translocation of cytosolic subunits, and de novo protein synthesis. These novel findings provide insight into the molecular regulation of NAD(P)H oxidase by Ang II in HVSMCs. Furthermore, we identify differences in gp91phox homologue expression in VSMCs from rats and human small and large arteries.

[Effects of grape seed proanthocyanidins extracts on experimental diabetic nephropathy in rats].

OBJECTIVE: To study the effects of grape seed proanthocyanidins extracts(GSPE) and its mechanism on early renal lesions of diabetic rats. METHODS: Diabetic rats induced by alloxan were given GSPE intragastrically for 6 weeks, then the antioxidative indexes and NO content, NOS activity in kidney and serum were measured, and the renal function indexes were tested as well. RESULTS: Compared with the diabetic group, the urinary protein/24h, levels of blood urea nitrogen(BUN)and serum creatinine(SCr), creatinine clearance rate(CCr) and the ratio of kidney weight/body weight were decreased, the SOD activity in kidney was raised while MDA content was fall in the GSPE group(high dose), and the differences were all significant. The NO content in the kidney and NOS activity in kidney and serum decreased in the GSPE (low dose)group, and there was significant difference when compared with diabetic group( P <0.05) . CONCLUSION: GSPE has the effect in protecting kidney of diabetic rats, the mechanism might be related with its action in increasing the renal antioxidative ability,decreasing the content of NO and the activity of NOS in kidney and serum.

Negative regulation of RhoA/Rho kinase by angiotensin II type 2 receptor in vascular smooth muscle cells: role in angiotensin II-induced vasodilation in stroke-prone spontaneously hypertensive rats.

OBJECTIVE: To test whether angiotensin II (Ang II) through the Ang II type 2 receptor (AT2R), downregulates RhoA/Rho kinase, which plays a role in AT1 receptor (AT1R)-mediated function. METHODS: In vitro studies were performed in A10 vascular smooth muscle cells (VSMC) and in vivo studies in mesenteric arteries from Wistar-Kyoto (WKY) and stroke-prone spontaneously hypertensive (SHRSP) rats. VSMC were stimulated with Ang II (10 mol/l), CGP42112A (10 mol/l, a selective AT2R agonist) +/- valsartan (10 mol/l, an AT1R antagonist), or the Rho kinase inhibitor fasudil (10 mol/l). AT1R and AT2R expression and myosin light chain (MLC) phosphorylation were determined by immunoblotting. RhoA activity was assessed by measuring membrane translocation. Functional significance between AT2R, RhoA/Rho kinase and vasodilation was assessed in arteries from valsartan-treated (30 mg/kg per day, 14 days) WKY and SHRSP rats. Vasodilatory responses to Ang II (10-10 mol/l) were performed in norepinephrine pre-contracted vessels +/- valsartan(10 mol/l), PD123319 (10 mol/l, an AT2R antagonist) or fasudil (10 mol/l). RESULTS: A10 VSMC expressed AT1R and AT2R. In valsartan-treated cells, Ang II-induced RhoA translocation was reduced versus controls (42 +/- 6%, P < 0.05). Similar responses were obtained with CGP42112A (45 +/- 6%, P < 0.05). This was associated with decreased MLC activation. Fasudil abrogated Ang II- and CGP42112A-mediated effects. Ang II evoked a significant vasodilatory response only in valsartan-treated SHRSP (max dilation 40 +/- 7%). PD123319 blocked these effects. Fasudil increased AngII-induced relaxation in SHRSP vessels. AT2R expression was increased by valsartan (two- to three-fold) in SHRSP arteries. RhoA translocation was increased two-fold in untreated SHRSP (P < 0.05) and was reduced by valsartan (P < 0.05). These changes were associated with decreased MLC phosphorylation. CONCLUSIONS: Ang II/AT2R negatively regulates vascular RhoA/Rho kinase/MLC phosphorylation. These processes may play a role in Ang II-mediated vasodilation in conditions associated with vascular AT2R upregulation, such as in SHRSP chronically treated with AT1R blockers, which may contribute to blood pressure lowering by these antihypertensive agents.

Vascular smooth muscle cell NAD(P)H oxidase activity during the development of hypertension: Effect of angiotensin II and role of insulinlike growth factor-1 receptor transactivation.

OBJECTIVE: We investigated whether angiotensin II (Ang II)-induced reactive oxygen species (ROS) generation is altered in vascular smooth muscle cells (VSMCs) from spontaneously hypertensive rats (SHR) during the phases of prehypertension, developing hypertension, and established hypertension and assessed the putative role of insulinlike growth factor-1 receptor (IGF-1R) in Ang II-mediated actions. METHODS: The VSMCs from SHR and Wistar-Kyoto rats (WKY) aged 4 (prehypertensive), 9 (developing hypertension), and 16 (established hypertension) weeks were studied. The ROS production and NAD(P)H oxidase activation were determined by fluorescence and chemiluminescence, respectively. The role of IGF-1R was assessed with the selective inhibitor AG1024. The ROS bioavailability was manipulated with Tiron (10(-5) mol/L) and diphenylene iodonium (DPI) (10(-6) mol/L). RESULTS: Angiotensin II dose dependently increased ROS production in WKY and SHR at all ages. The Ang II-induced responses were greater in SHR versus WKY at 9 and 16 weeks (P < .05). The Ang II-stimulated ROS increase was greater in 9- and 16-week-old SHR versus 4-week SHR (P < .05). These effects were reduced by AG 1024. Basal NAD(P)H oxidase activity was higher in VSMCs from 9-week-old SHR versus 4-week-old rats (P < .05). Angiotensin II induced a significant increase in oxidase activity in VSMCs from 9- and 16-week-old SHR (P < .001), without influencing responses in cells from 4-week-old SHR. Pretreatment of 9- and 16-week-old SHR cells with AG1024 reduced Ang II-mediated NAD(P)H oxidase activation (P < .05). CONCLUSIONS: Basal and Ang II-induced NAD(P)H-driven ROS generation are enhanced in VSMCs from SHR during development of hypertension, but not in cells from prehypertensive rats. Transactivation of IGF-1R by Ang II may be important in vascular oxidative excess in the development of hypertension in SHR.

[Studies of the norm and psychometrical properties of the ages and stages questionnaires, third edition, with a Chinese national sample].

OBJECTIVE: To introduce the Ages and Stages Questionnaires, Third Edition (ASQ-3), to China, created ASQ-Chinese (ASQ-C) and carried out studies of its national norm and the psychometrical properties in the children aged 1-66 months in the mainland of China in collaboration with the author of the ASQ System and under the authorizations from its publisher on translation, researches, publication and distribution of the ASQ-3. METHOD: The ASQ-3 questionnaires were translated and adapted into a Simplified Chinese version, the ASQ-C, with six steps such as translation, back-translation and adaptation and so on to ensure consistency with the core of the original document and to have the cultural relevance in China.A stratified cluster sampling method was utilized to recruit children aged 1-66 months with respect to demographic characteristics such as the proportion of population in each administrative region and in urban and rural areas and so on that are representative of 2010 China census data.A sample size of over 200 was collected for each ASQ-C age interval.Children were excluded from the normative sample who (1) are from communities or villages at an elevation of 2 000 m or above and(or) where simplified Chinese is not the official language, or (2) had been diagnosed as having a developmental delay by any authoritative organizations.The national normative sample for the ASQ-C had a total sample size of 4 452, sample size within each age interval ranged from 218 to 227, including 2 230 male cases and 2 222 female cases, 2 236 urban cases and 2 216 rural cases.A convenience sample was recruited from the normative sample to examine inter-rater reliability and test-retest reliability in all six administrative regions.Researchers completed the ASQ-C on the same child with their parents for 162 children for inter-rater reliability(the size of each ASQ-C age interval was 5-9); parents of 168 children completed another age-appropriate ASQ-C for test-retest reliability during 10-15 days after they completed the normative ASQ-C(The size of each ASQ-C age interval is 6-10). Another convenience sample was recruited from the follow-up of low birth weight infants for the concurrent validity of the ASQ-C in comparison with the Beijing Gesell.Parents of 198 children completed age-appropriate ASQ-C and professional administered to the children with the Beijing Gesell.In the ASQ-C norm and test-retest reliability, parents completed the age-appropriate ASQ-C, independently or with needed assistance. In inter-rater reliability, researchers completed the same ASQ-C after parents. In validity test, after parents completing age-appropriate ASQ-C, professional tested children with the Beijing Gesell.Data were analyzed using SPSS version 13.0 software.The mean and standard deviation of the national normative sample were calculated, reliability and validity of the ASQ-C was examined. RESULT: The demographic characteristics of this Chinese sample match the 2010 China census data on gender, urban or rural location, and family income.All 20 intervals of the ASQ-C were standardized on 21 national normative samples.Cronbach's alpha coefficient for the whole measure was 0.8.The Pearson correlation coefficient between the ASQ-C total scores of the two raters was 0.8.The Pearson correlation coefficient between the ASQ-C total scores of the two times was 0.8 (all P<0.000 1). The sensitivity of ASQ-C was 87.50% and the specificity of ASQ-C was 84.48%.The percentage of the agreement between the ASQ-C and the Beijing Gesell was 84.74%. CONCLUSION: These findings indicate that the ASQ-C is a reliable and valid measure with a representative national sample aged 1-66 months.It can be used to screen and monitor the development of children in the mainland of China.

Inhibitors of Na+/Mg2+ exchange activity attenuate the development of hypertension in angiotensin II-induced hypertensive rats.

OBJECTIVES: To investigate whether imipramine and quinidine, inhibitors of the Na /Mg exchanger, influence development of hypertension in rats infused with angiotensin (Ang) II. METHODS: Sprague-Dawley rats were divided into six groups: (1) control (vehicle); (2) Ang II (150 ng/kg per min subcutaneously); (3) imipramine alone (5 mg/kg per day in drinking water); (4) quinidine alone (5 mg/kg per day in drinking water); (5) Ang II plus imipramine; (6) Ang II plus quinidine. Rats were studied for 3 weeks. To verify that Ang II directly influences Na -dependent Mg exchange, in-vitro studies were performed in vascular smooth muscle cells (VSMCs) derived from mesenteric arteries. RESULTS: Ang II increased systolic blood pressure (SBP) in all groups. The magnitude of the increase was lower ( 0.01) in Ang II groups treated with imipramine (151 +/- 7.4 mmHg) or quinidine (163 +/- 4 mmHg) than in the Ang II only group (205 +/- 4 mmHg). Neither imipramine nor quinidine influenced SBP in vehicle-treated rats. Plasma concentrations of Mg and K were decreased in Ang II rats compared with controls (P < 0.05). Platelet intracellular free Mg concentration was reduced and platelet intracellular free Na concentration was increased in the Ang II group compared with control and treated groups (P < 0.01). These effects were normalized by imipramine and quinidine. Ang II stimulated Na -dependent Mg transport in VSMCs. These actions were abrogated by imipramine and quinidine and in Na -free conditions. CONCLUSIONS: Our data demonstrate that inhibitors of Na -dependent Mg transport attenuate development of hypertension in rats infused with Ang II. These findings suggest a possible role for Na /Mg exchange activity in the pathogenesis of Ang II-dependent hypertension.

Effects of low dietary magnesium intake on development of hypertension in stroke-prone spontaneously hypertensive rats: role of reactive oxygen species.

OBJECTIVES: To investigate whether low dietary Mg2+ intake influences the development of hypertension in stroke-prone spontaneously hypertensive rats (spSHRs) and whether these effects are associated with vascular functional and structural changes, and to assess the role of reactive oxygen species and the activation of vascular mitogen-activated protein (MAP) kinases in these processes. METHODS: Six-week-old male spSHRs (n = 18) were divided into three groups: control (normal chow, 0.21% Mg2+ ), low Mg2+ group (Mg2+ -free diet), and high Mg2+ group (Mg2+ -rich diet, 0.75%). Systolic blood pressure (SBP) was assessed weekly for 16 weeks. In a second series of experiments, 6-week-old spSHRs (n = 18) were divided into three groups and studied weekly for 7 weeks: control group, low Mg2+ group, and low Mg2+ group receiving the superoxide dismutase mimetic, tempol (1 mmol/l). RESULTS: The low Mg2+ diet caused an initial decrease in SBP followed, 5 weeks later, by an exacerbated development of hypertension. This was associated with a transient reduction in the plasma concentrations of substances associated with the thiobarbituric acid reaction (markers of oxidative stress), which increased rapidly 2 weeks later. In the low Mg2+ group, acetylcholine-induced vasodilatation was decreased compared with that in controls ( P<0.05). The media : lumen ratio was greater in rats receiving a low Mg2+ diet than in those fed a high Mg2+ diet ( P<0.05). Mg2+ depletion was associated with increased vascular superoxide anion compared with that in Mg2+ -supplemented rats (1.2 0.24 compared with 0.65 0.1 nmol/min per mg). Phosphorylation of MAP kinases was increased two- to threefold in Mg2+ -deficient rats. Tempol prevented the progression of hypertension and normalized the vascular changes in rats fed a low Mg2+ diet. CONCLUSIONS: Chronic Mg2+ deficiency leads to development of severe hypertension, endothelial dysfunction and vascular remodelling. These processes are associated with oxidative stress and upregulation of redox-dependent MAP kinases. Tempol normalized vascular changes and attenuated the development of hypertension. Our findings suggest that reactive oxygen species play an important part in vascular processes that are associated with progression of hypertension in Mg2+ -deficient spSHRs.

Angiotensin II and endothelin-1 regulate MAP kinases through different redox-dependent mechanisms in human vascular smooth muscle cells.

OBJECTIVE: The role of reactive oxygen species (ROS) in mitogen-activated protein kinase (MAPK) signaling by angiotensin (Ang) II and endothelin-1 (ET-1) in human vascular smooth muscle cells (VSMC) was investigated. DESIGN: VSMCs were derived from resistance arteries from healthy subjects. MAPK activity was assessed using phospho-specific antibodies. ROS generation was measured by CMH2DCFDA fluorescence and nicotinamide adenine dinucleotide phosphate (NADPH) oxidase activity by lucigenin chemiluminescence. RESULTS: Ang II and ET-1 increased MAPK phosphorylation (P < 0.01). Pre-treatment with Tiron and Tempol, *O2 scavengers, attenuated agonist-stimulated phosphorylation of p38MAPK, c-Jun N-terminal kinases (JNK) and ERK5, but not of ERK1/2 (extracellular signal-regulated kinases). Apocynin and diphenylene iodinium (DPI), NAD(P)H oxidase inhibitors, decreased Ang II-induced responses 60-70%. ET-1-mediated MAPK phosphorylation was unaffected by apocynin but was reduced (> 50%) by thenoyltrifluoroacetone (TIFT) and carboxyl cyanide-m-chlorophenylhydrazone (CCCP), mitochondrial inhibitors. Allopurinol and N-nitro-l-arginine methyl ester (l-NAME), xanthine oxidase and nitric oxide synthase (NOS) inhibitors, respectively, did not influence MAPK activation. Intracellular ROS generation, was increased by Ang II and ET-1 (P < 0.01). DPI inhibited Ang II- but not ET-1-mediated ROS production. Expression of p22phox and p47phox and activation of NAD(P)H oxidase were increased by Ang II but not by ET-1. CCCP and TIFT significantly attenuated ET-1-mediated ROS formation (P < 0.05), without influencing Ang II effects. CONCLUSIONS: Ang II activates p38MAPK, JNK and ERK5 primarily through NAD(P)H oxidase-generated ROS. ET-1 stimulates these kinases via redox-sensitive processes that involve mitochondrial-derived ROS. These data suggest that redox-dependent activation of MAPKs by Ang II and ET-1 occur through distinct ROS-generating systems that could contribute to differential signaling by these agonists in VSMCs.

Effect of synthetic link N peptide on the expression of type I and type II collagens in human intervertebral disc cells.

Intervertebral disc (IVD) degeneration is associated with proteolytic degradation of proteoglycan aggregates present within the extracellular matrix of the disc. Link N peptide (DHLSDNYTLDHDRAIH) is the N-terminal peptide of link protein, which stabilizes the proteoglycan aggregates. It is generated in vivo by proteolytic degradation during tissue turnover. It has been previously shown that this peptide can stimulate the synthesis of collagens by articular cartilage and bovine IVD cells in vitro. Being a synthetic peptide, Link N has considerable financial benefits for clinical use over recombinant growth factors because it is extremely cheap to produce. The purpose of the present study was to determine the effect of Link N on the expression of types I and II collagen and investigate the cellular mechanisms of Link N signal transduction in human IVD cells. The present results suggest that Link N stimulates the expression of types I and II collagen in human IVD cells. More specifically, Link N stimulated the expression of type I in nucleus pulposus (NP) cells, but not in annulus fibrosus cells. As Link N also decreased the phosphorylation of p38 in NP cells only, results suggest that p38 is a mediator of the effect of Link N on type I collagen expression. p38 is a member of the mitogen-activated protein kinase family highlighted by three major cascades: p38, c-Jun amino-terminal kinase, and extracellular signal-regulated kinase pathways. Link N showed no effect on the latter two pathways, suggesting a specific effect of Link N on the p38 cascade. On the other hand, Link N stimulated the expression of type II collagen in both NP and annulus fibrosus, suggesting that other mechanisms are implicated in the control of type II collagen expression in disc cells, without excluding p38 for the NP. In conclusion, the present study showed that Link N can modulate the expression of collagen in human IVD cells.

Effect of parathyroid hormone on type X and type II collagen expression in mesenchymal stem cells from osteoarthritic patients.

A major drawback of current cartilage and intervertebral disc tissue engineering is that human mesenchymal stem cells (MSCs) from osteoarthritic (OA) patients express type X collagen (COL10), a marker of late-stage chondrocyte hypertrophy (associated with endochondral ossification). Parathyroid hormone (PTH) regulates endochondral ossification by inhibiting chondrocyte differentiation toward hypertrophy. In this study, we investigated the effect of PTH on expression of COL10 in MSCs from OA patients and analyzed the potential mechanisms related to its effect. MSCs were obtained from aspirates from the intramedullary canal of donors undergoing total hip replacement for OA. Expanded cells were then incubated for 0-48 h without (control) or with 100 nM PTH (1-34). Protein expression and phosphorylation were measured by Western blot. Results showed that PTH (1-34) inhibited expression of COL10 in MSCs from OA patients in a time-dependent manner. In parallel, PTH (1-34) stimulated expression of COL2, a marker of chondrogenic differentiation. Results also showed that PTH (1-34) inhibited in a sustained manner the phosphorylation of p38 and AKT protein kinase signaling pathways. Interestingly, the modulation of COL2 and COL10 gene expression was significant as rapidly as after 1 h in the presence of PTH (1-34); changes in the phosphorylation of p38 and AKT were significant only after 6 h. This suggests that while p38 and AKT protein kinase signaling pathways may not be required to initiate the regulation of expression of COL2 and COL10 by PTH (1-34), these pathways may modulate later events necessary for preventing precocious MSC hypertrophy.