Myelin regulatory factor ( Myrf ) is a critical early regulator of retinal pigment epithelial development.

Myelin regulatory factor (Myrf) is a critical transcription factor in early retinal and retinal pigment epithelial development, and human variants in MYRF are a cause for nanophthalmos. Single cell RNA sequencing (scRNAseq) was performed on Myrf conditional knockout mice ( Rx>Cre Myrf fl/fl ) at 3 developmental timepoints. Myrf was expressed specifically in the RPE, and expression was abrogated in Rx>Cre Myrf fl/fl eyes. scRNAseq analysis revealed a loss of RPE cells at all timepoints resulting from cell death. GO-term analysis in the RPE revealed downregulation of melanogenesis and anatomic structure morphogenesis pathways, which were supported by electron microscopy and histologic analysis. Novel structural target genes including Ermn and Upk3b , along with macular degeneration and inherited retinal disease genes were identified as downregulated, and a strong upregulation of TGFß/BMP signaling and effectors was observed. Regulon analysis placed Myrf downstream of Pax6 and Mitf and upstream of Sox10 in RPE differentiation. Together, these results suggest a strong role for Myrf in the RPE maturation by regulating melanogenesis, cell survival, and cell structure, in part acting through suppression of TGFß signaling and activation of Sox10 . SUMMARY STATEMENT: Myrf regulates RPE development, melanogenesis, and is important for cell structure and survival, in part through regulation of Ermn , Upk3b and Sox10, and BMP/TGFb signaling.

Advanced maternal age impairs spatial learning capacity in young adult mouse offspring.

Effects of maternal aging on the offspring cognitive function remain controversial in population-based investigations, and information available in animal studies is very limited. We investigated the impact of a delayed first natural pregnancy on pregnancy outcomes in the mouse model. Spatial learning capacity in young adult mouse offspring was observed by step-down passive avoidance task and Morris water maze (MWM). Maternal serum α-klotho was measured by ELISA. Morphological characteristics of fetoplacental unit and offspring brain were identified by H&E and immunohistochemistry. Klotho, VDR and other related genes expression were quantified by real-time-RT-PCR and western blot. We found delayed pregnancy reduced fertility in female mice by three-fold (Young vs. Old: 5.0% vs. 20.7%), and increased adverse pregnant outcomes by eight-fold (Young vs. Old: 3.0% vs. 27.5%). Mice born to old mothers exhibited shorter retention trial latency in passive avoidance task and longer latency to find the platform in MWM, suggesting worse performance on the tests that measure learning and memory. Serum α-klotho level was lower in old female mice before pregnancy, whereas became comparable after pregnancy. Vitamin D receptor (VDR) expression, both in mRNA and protein, markedly decreased during the early stage of fetoplacental unit in old mice, especially in trophoblast giant cells when compared with that of young mice. Importantly, consistent with fetoplacental unit, VDR expression also declined in hippocampus from offspring born to old mice. These results suggest that young adult offspring from aged mothers exhibited worse cognitive function and the reduced VDR expression during fetoplacental development might play an important role.

[Tetramethoxystilbene, a selective CYP1B1 inhibitor, suppresses adipogenesis of C3H10T1/2 pluripotent stem cells].

OBJECTIVE: To investigate the inhibitory effects of tetramethoxystilbene, a selective CYP1B1 inhibitor, on adipogenic differentiation of C3H10T1/2 multi-potent mesenchymal cells. METHODS: In vitro cultured C3H10T1/2 cells at full confluence were induced by adipogenic agents (10 µg/ml insulin, 2 µmol/L dexamethasone and 0.5 mmol/L 3-isobutyl-1-methylxanthine) and exposed simultaneously to TMS at the final concentrations of 1.0, 2.0 or 4.0 µg/ml. Oil Red-O staining was used to observe the cell differentiation. The expression of peroxisome proliferator-activated receptor gamma (PPARγ) and its target genes cluster of differentiation 36 (CD36) and fatty acid binding protein 4 (FABP4) were quantified by real-time RT-PCR and Western blotting. RESULTS: Oil Red-O staining and TG contents revealed that TMS suppressed induced differentiation of C3H10T1/2 cells. TMS exposure of the cells dose-dependently decreased both mRNA and protein expressions of PPARγ, a key nuclear transcription factor during adipogenesis, and also lowered the mRNA expressions of PPARγ target genes CD36 and FABP4. CONCLUSION: TMS can suppress adipogenic differentiation of C3H10T1/2 cells by inhibiting PPARγ

Determinants of metabolic syndrome in Chinese schoolchildren.

The objective of this study was to identify the potential risk factors of metabolic syndrome (MetS) among Chinese schoolchildren. A cross-sectional study among 624 children (357 boys and 267 girls, aged 9.6 ± 0.7 years) was conducted in Wuhan, China, from May to June 2010. MetS was defined according to the criteria proposed by De Ferranti and the International Diabetes Federation (IDF) criteria. Data on cardiorespiratory fitness (CRF), household income, parental hypertension, and children's personal information, including birth weight, preterm birth, and breast-feeding, reported by their parents were obtained. Multiple logistic regression showed that CRF (odds ratio [OR] = 0.68; 95% confidence interval [CI] = 0.60-0.77), breast-feeding (OR = 0.32; 95% CI = 0.10-0.97), and paternal hypertension (OR = 5.06; 95% CI = 1.20-21.37) were all independently associated with MetS. In conclusion, low CRF and paternal hypertension significantly increase the risk, whereas breast-feeding may reduce the risk of MetS among Chinese schoolchildren.

Association between alcohol consumption and risk of cardiovascular disease and all-cause mortality in patients with hypertension: a meta-analysis of prospective cohort studies.

OBJECTIVE: To conduct a meta-analysis summarizing the risk of cardiovascular disease (CVD) and all-cause mortality (ACM) in relation to alcohol consumption in patients with hypertension, focusing on clarifying dose-response associations. PATIENTS AND METHODS: PubMed and EMBASE were searched for eligible prospective cohort studies from December 3, 1949, through January 18, 2014. The semi-parameter method and dose-response analysis were used. RESULTS: Nine studies (11 cohorts) were included in the meta-analysis. Compared with the lowest alcohol level (abstainers/occasional drinkers), the pooled relative risk (RR) was 0.72 (95% CI, 0.68-0.77) for the third highest category (median, 10 g/d), 0.81 (95% CI, 0.71-0.93) for the second highest category (median, 20 g/d), and 0.60 (95% CI, 0.54-0.67) for the highest category (median, 30 g/d). A J-shaped relationship between alcohol use and ACM was observed, and the nadir (RR, 0.82; 95% CI, 0.76-0.88) was found to be at a dose of 8 to 10 g of alcohol consumption per day. CONCLUSION: Findings of this meta-analysis suggest that low-to-moderate alcohol consumption was inversely significantly associated with the risk of CVD and ACM in patients with hypertension.

Fuzheng Qingjie recipe induces apoptosis in HepG2 cells via P38 MAPK activation and the mitochondria-dependent apoptotic pathway.

Fuzheng Qingjie (FZQJ) recipe is a polyherbal Chinese medicine capable of suppressing tumor growth and is used as an adjuvant therapy for various types of cancer. However, its anticancer mechanisms are yet to be fully elucidated. In the present study, we explored whether p38 mitogen-activated protein kinase (MAPK) was involved in FZQJ-mediated mitochondria-dependent apoptosis in human hepatocellular carcinoma cells. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays were used to measure the viability of HepG2 cells. 4,6-Diamidino-2-phenylindole (DAPI) and Annexin-V fluorescein isothiocyanate (FITC) were used to analyze the apoptosis of HepG2 cells. The mitochondrial membrane potential (∆ψ) and phosphorylated P38 MAPK protein were examined by a flow cytometer following 5,5',6,6'-tetrachloro­1,1',3,3'-tetraethylbenzimidazolcarbocyanine iodide (JC-1) and Alexa Fluor® 647 mouse anti-phosphorylated P38 MAPK antibody staining, respectively. The activation of caspase-9 and caspase-3 were measured using colorimetric assays. Additionally, Bcl-2 and Bax expression were examined using reverse transcription polymerase chain reaction (RT-PCR) and western blot analysis. The results demonstrated that water extract of FZQJ was able to induce apoptosis of HepG2 cells in vitro. FZQJ-induced apoptosis was accompanied by the loss of ∆ψ, downregulation of Bcl-2 and upregulation of Bax expression, and the activation of caspase-3, -9 and P38 MAPK. These results indicated that FZQJ induced apoptosis in HepG2 cells at least via P38 MAPK activation and the mitochondria-dependent apoptotic pathway.

[Effects of preconditioning of wufu jingfang on ischemia/reperfusion injury induced apoptosis in rats].

OBJECTIVE: To observe the protective effect of Wufu Jingfang (WJ, containing Aconitum carmichaeli Debx, Radix Aconiti Lateralis Preparatae, Rhizoma Pinelliae, and snakegourd fruit) on myocardial ischemia-reperfusion injury (I/R) of rats, thus exploring the feasibility of recipes containing eighteen incompatible pairs for specific pathological conditions. METHODS: Fifty male Wistar rats were randomly divided into five groups, i.e., the sham-operative control group (the SH group), the I/R group, the low dose WJ I/R group (the I/R +JFL group), the middle dose WJ I/R group (the I/R +JFM group), the high dose WJ I/R group (the I/R +JFH group), 10 in each group. Rats in the latter three groups were administered with WJ at 0.75 mL/100 g, 1.50 mL/100 g, and 3.00 mL/100 g body weight for 14 consecutive days by gastrogavage. All groups except the SH group received ligation of left anterior descending branch of coronary artery for 30-min ischemia followed by 120-min reperfusion. The micro-structural changes of myocardial mitochondria were observed by transmission electron microscope. The ischemic cardiomyocyte apoptosis was detected in each group using one-step terminal-deoxynucleotidyl transferase mediated nick end labeling (TUNEL). The mRNA expressions of B-cell leukemia/lymphoma 2 (Bcl-2) and Bcl-2 associated x protein (Bax) were detected by RT-PCR. The activities of lactic dehydrogenase (LDH) and creatine kinase (CK) were detected using ELISA. The myocardial infarct size was detected. RESULTS: Compared with the I/R group, WJ pretreatment significantly suppressed the release of LDH and CK (Besides, the release of LDH and CK reduced along with increased dose.), reduced the myocardial infarct size, and lowered myocardial apoptosis index (P < 0.05). WJ pretreatment also modulated Bcl-2/Bax ratio by up-regulating Bcl-2 expression level while decreasing Bax expression level. CONCLUSIONS: WJ pretreatment might protect the heart from I/R injury via decreasing myocardial cell apoptosis. The results suggested that eighteen incompatible pairs is not absolute, but relative. Chinese medical preparation containing opposite Chinese herbs could be used in specific pathological states such as ischemic cardiomyopathy.

Physical Activity and Lung Function Growth in a Cohort of Chinese School Children: A Prospective Study.

BACKGROUNDS/OBJECTIVES: Evidence on the association between physical activity and lung function in children is sparse. The aim of this study was to evaluate children's lung function growth in relation to their physical activity level in Chinese children. METHODS: A total of 1713 school children aged 9.89±0.86 years who were asthma-free at baseline were followed-up for 18 months from 2006 to 2008 in Guangzhou, China. Information on physical activity and other socio-economic status were obtained from self-administered questionnaires. Lung function tests were performed with a standard procedure. RESULTS: At the baseline survey, physically active girls had significantly higher forced vital capacity (FVC) than inactive girls (1.79 l vs. 1.75 l, p<0.05). The growth rates for lung function indices were significantly higher for girls who were physically active at either or both follow-up surveys than those inactive at both surveys during the follow-up period forced expiratory flows at 25% (FEF25) difference per year (dpy) (0.20 l/s vs. 0.15 l/s), forced expiratory flows at 75% (FEF75) dpy (0.57 l/s vs. 0.45 l/s) and forced expiratory flows between 25% and 75% (FEF25-75) dpy (0.36 l/s vs. 0.28 l/s) (all p<0.05). CONCLUSIONS: Physical activity is positively associated with lung function growth among Chinese school-aged girls. Promotion of physical activity among children is of great importance.

4-Benzyl-3,5-dimethyl-1H-pyrazole.

In the title mol-ecule, C(12)H(14)N(2), the dihedral angle between the pyrazole and phenyl ring mean planes is 78.65 (19)°. In the crystal, mol-ecules are linked by N-H⋯N hydrogen bonds into chains along [010].

Relationship of body fat and cardiorespiratory fitness with cardiovascular risk in Chinese children.

UNLABELLED: BACKGROUNDS/OBJECTIVES: Cardiorespiratory fitness (CRF) and body fat play an important role in elevated risk for cardiovascular disease (CVD). However, the combined effects of CRF and obesity on metabolic health in Chinese children are unclear. The purpose of this study was to investigate the independent and combined associations between body fat, CRF, and CVD risk in Chinese schoolchildren. METHODS: The study subjects comprised 676 schoolchildren (392 boys and 284 girls, aged 9.6±0.7 yrs old) in Wuhan, China. Their body mass index (BMI), waist circumference (WC), CRF, blood pressure (BP), lipids, glucose, and pubertal status were assessed. Children were categorized into different groups based on their BMI, WC, and CRF using Chinese obesity cut-off points and CRF sex-specific median points. Metabolic Risk Score (MRS) was computed based on the standardized scores of BP, lipids, and glucose. RESULTS: Multiple linear regression models showed that, in the separate models, body fat was positively associated with MRS while CRF was inversely associated with MRS (p<0.001). However, when assessed simultaneously, only body fat had a significant association with MRS (p<0.001). In general, low-fit children had a lower MRS compared to their counterparts, and a significant difference between the two extreme groups was observed (low CRF and high fat vs. high CRF and low fat, p<0.001). CONCLUSIONS: These findings suggest that both body fat and CRF should be considered when interpreting CVD risk in Chinese children, while body fat may be correlated with CVD risk more than CRF.

Tetra-kis(3,5-dimethyl-1H-pyrazole-κN)(nitrato-κO,O')cadmium(II) nitrate.

The title compound, [Cd(NO(3))(C(5)H(8)N(2))(4)]NO(3), was prepared by reaction of cadmium nitrate and 3,5-dimethyl-pyrazole in ethanol solution. The Cd atom adopts a distorted cis-CdO(2)N(4) octa-hedral geometry involving four dimethylpyrazole molecules and one bidentate nitrate anion. The mol-ecular structure and packing are stabilized by N-H⋯O and C-H⋯O inter- and intra-molecular hydrogen-bonding inter-actions.

N-[(3,5-Dimethyl-pyrazol-1-yl)meth-yl]phthalimide.

The title compound {systematic name: 2-[(3,5-dimenthylpyrazol-1-yl)meth-yl]isoindole-1,3-dione}, C(14)H(13)N(3)O(2), was prepared by reaction of N-(bromo-meth-yl)phthalimide and 3,5-dimethyl-pyrazole in chloro-form solution. The mol-ecular structure and packing are stabilized by intra-molecular C-H⋯O hydrogen-bonding and C-H⋯π inter-actions.

N-(Imidazol-1-ylmeth-yl)phthalimide.

THE TITLE COMPOUND [SYSTEMATIC NAME: 2-(imidazol-1-ylmeth-yl)isoindole-1,3-dione], C(12)H(9)N(3)O(2), was prepared by reaction of N-(bromo-meth-yl)phthalimide and imidazole in chloro-form solution. The crystal structure is stabilized by weak inter-molecular C-H⋯π inter-actions and inter-molecular π-π inter-actions with centroid-centroid distances in the range 3.6469 (8)-3.8831 (9) Å.

N-(1H-1,2,3-Benzotriazol-1-ylmeth-yl)phthalimide.

THE TITLE COMPOUND [SYSTEMATIC NAME: 2-(1H-1,2,3-benzotriazol-1-ylmeth-yl)isoindole-1,3-dione], C(15)H(10)N(4)O(2), was prepared by the reaction of 1H-benzotriazole and 2-bromo-methyl-isoindole-1,3-dione. The benzotriazole and isoindole units are almost planar and make a dihedral angle of 70.2 (1)° (mean planes include C and N atoms). A weak C-H⋯O intra-molecular hydrogen bond involving a carbonyl O atom as acceptor stabilizes the observed mol-ecular conformation.

[Brain-derived neurotrophic factor mRNA expressions in rat brain induced by short-term in vivo exposure to deltamethrin].

OBJECTIVE: To elucidate the mechanism of damage on central nervous system (CNS) caused by deltamethrin (DM). METHODS: The mRNA and protein expressions of brain-derived neurotrophic factor (BDNF) in the cerebral cortex and hippocampus of the rats exposed to DM were measured by retro-transcription-polymerase chain reaction (RT-PCR), dot blot, flow cytometry analysis and immunohistochemistry. RESULTS: After exposure to DM at high-dose (DM1, 25.0 mg x kg(-1) x d(-1), i.p.) once and low-dose (DM2, 12.5 mg x kg(-1) x d(-1), i.p.) for 5 days, the level of BDNF mRNA and protein expression in the cerebral cortex and hippocampus of the rats increased significantly. The levels of BDNF mRNA and protein expression in the cerebral cortex and hippocampus measured by of RT-PCR in the rats with DM1 and DM2 were higher than those in the controls by 48% and 56%, and 59% and 54%, respectively. And, those measured by dot blot in the rats with MD1 and MD2 were 186% and 161%, and 148% and 158% of those in the controls, respectively, basically similar to those measured by RT-PCR. Flow cytometric analysis showed that the levels of BDNF mRNA and protein expression in the cerebral cortex and hippocampus in the rats with DM1 and DM2 were higher than those in the controls by 53% and 89%, and 45% and 46%, respectively. Immunohistochemical analysis showed that protein expression in the cerebral cortex of the rats with DM1 and DM2 were 129% and 147% of those in the controls, same as the flow cytometric analysis, but those were significantly higher in the hippocampus mainly in the CA1 and DG areas of the rats with MD1 and the CA3 and DG areas of the rats with DM2. CONCLUSIONS: DM could induce BDNF mRNA and protein expression in the cerebral cortex and hippocampus of the rats, which could play an important role in repairing of nerve damage.

[Effects of deltamethrin on cell survival rate and intracellular free Ca2+ concentration in primary cultured astrocytes of rat].

OBJECTIVE: To study the effects of deltamethrin (DM) on cell survival rate and intracellular Ca(2+) ([Ca(2+)]i) concentration in primary cultured astrocytes of rat. METHODS: The cell survival rate was measured by Typan Blue assay; the intracellular [Ca(2+)]i concentration was determined by the fluorescent Ca(2+) indicator Fura-2/AM. RESULTS: The survival rate of astrocytes was decreased to 91.9% after astrocytes were incubated with 1 x 10(-5) mol/L DM for 72 h (P < 0.05). The cell survival rates were 89.0%, 84.8%, 81.2% and 79.2% respectively when astrocytes were administered with 1 x 10(-4) mol/L DM for 4, 12, 24 and 72 h, which were remarkably lower than control groups (P < 0.01). Comparing with controls and before DM treatment, sharp increases in [Ca(2+)]i concentration [(451.4 +/- 42.3), (536.9 +/- 47.5) and (870.9 +/- 100.5) nmol/L respectively] were observed when astrocytes were incubated with 1 x 10(-7), 1 x 10(-6) and 1 x 10(-5) mol/L DM for 5 minutes (P < 0.01). After astrocytes were treated with 1 x 10(-8), 1 x 10(-7), 1 x 10(-6), 1 x 10(-5) mol/L DM for 15 minutes, the [Ca(2+)]i concentrations were decreased to (124.3 +/- 6.0), (131.3 +/- 19.1), (118.9 +/- 1.4), (136.6 +/- 3.8) nmol/L respectively, which were significantly different from those of controls and before treatment. And this situation was almost keeping stable to 30 min. CONCLUSION: The cell survival rate was decreased and the [Ca(2+)]i concentration was temporarily increased when astrocytes were treated with DM.

[Effects of pyrethroids on the activity of gamma-aminobutyric acid transferase in rat brain].

OBJECTIVE: To study the effects of pyrethroids on the activity of gamma-aminobutyric acid transferase (GABAT) in rat brain. METHOD: The coupled enzyme ultraviolet spectrophotography was applied to observe the effects of deltamethrin (DM) and permethrin (PM) on the activities of GABAT in rat cerebral cortex, hippocampus, corpus striatum and cerebellum in vitro and in vivo. RESULTS: In vitro, DM and PM had no significant effects on the activities of GABAT in rat cerebral cortex, hippocampus, corpus striatum and cerebellum at the final concentration of 10(-9) - 10(-4) mol/L. When 37.5 mg/kg DM and 600 mg/kg PM were orally administrated to the rats at one time, the activities of GABAT in rat cerebral cortex, hippocampus and cerebellum in the DM group [(2.96 +/- 0.43), (2.13 +/-0.44), (5.12 +/- 1.36) nmol x mg pro(-1) x min(-1), respectively] were lower than those in the control group [(3.43 +/- 0.41), (2.68 +/- 0.47), (6.74 +/- 1.64) nmol x mg pro(-1) x min(-1)] (P < 0.05), and the activities of GABAT in rat cerebral cortex and hippocampus in the PM group [(4.57 +/- 0.30), (4.18 +/- 0.63) nmol.mg pro(-1) x min(-1), respectively] were higher than those in the control group (P < 0.05). When 12.5 mg/kg DM and 200 mg/kg PM were orally administrated to the rats once a day for consecutive five days, the two pesticides had no significant effects on the activities of GABAT in rat cerebral cortex, hippocampus, corpus striatum and cerebellum (P > 0.05). CONCLUSIONS: In vitro, DM and PM had no significant effects on the activity of GABAT in rat brain; in vivo, DM and PM may have different effects on the activity of GABAT in rat brain, which deserve further study.