Differential effects of sleep deprivation on behavior and microglia in a brain-region-specific manner in young and aged male mice.

Microglia, resident immune cells in the central nervous system, constantly monitor the state of the surrounding brain activity. The animal model induced by sleep deprivation (SD) is widely used to study the pathophysiological mechanisms of insomnia and bipolar disorder. However, it remains unclear whether SD affects behaviors in young and aged male mice and microglia in various brain regions. In this study, we confirmed brain region-specific changes in microglial density and morphology in the accumbens nucleus (Acb), amygdala (AMY), cerebellum (Cb), corpus callosum (cc), caudate putamen, hippocampus (HIP), hypothalamus (HYP), medial prefrontal cortex (mPFC), and thalamus (TH) of young mice. In addition, the density of microglia in old mice was higher than that in young mice. Compared with young mice, old mice showed a markedly increased microglial size, decreased total length of microglial processes, and decreased maximum length. Importantly, we found that 48-h SD decreased microglial density and morphology in old mice, whereas SD increased microglial density and morphology in most observed brain regions in young mice. SD-induced hyperactivity was observed only in young mice but not in old mice. Moreover, microglial density (HIP, AMY, mPFC, CPu) was significantly positively correlated with behaviors in SD- and vehicle-treated young mice. Contrarily, negative correlations were shown between the microglial density (cc, Cb, TH, HYP, Acb, AMY) and behaviors in vehicle-treated young and old mice. These results suggest that SD dysregulates the homeostatic state of microglia in a region- and age-dependent manner. Microglia may be involved in regulating age-related behavioral responses to SD.

Depletion of microglia with PLX3397 attenuates MK-801-induced hyperactivity associated with regulating inflammation-related genes in the brain.

Acute administration of MK-801 (dizocilpine), an N-methyl-D-aspartate receptor (NMDAR) antagonist, can establish animal models of psychiatric disorders. However, the roles of microglia and inflammation-related genes in these animal models of psychiatric disorders remain unknown. Here, we found rapid elimination of microglia in the prefrontal cortex (PFC) and hippocampus (HPC) of mice following administration of the dual colony-stimulating factor 1 receptor (CSF1R)/c-Kit kinase inhibitor PLX3397 (pexidartinib) in drinking water. Single administration of MK-801 induced hyperactivity in the open-field test (OFT). Importantly, PLX3397-induced depletion of microglia prevented the hyperactivity and schizophrenia-like behaviors induced by MK-801. However, neither repopulation of microglia nor inhibition of microglial activation by minocycline affected MK-801-induced hyperactivity. Importantly, microglial density in the PFC and HPC was significantly correlated with behavioral changes. In addition, common and distinct glutamate-, GABA-, and inflammation-related gene (116 genes) expression patterns were observed in the brains of PLX3397- and/or MK-801-treated mice. Moreover, 10 common inflammation-related genes ( CD68, CD163, CD206, TMEM119, CSF3R, CX3CR1, TREM2, CD11b, CSF1R, and F4/80) with very strong correlations were identified in the brain using hierarchical clustering analysis. Further correlation analysis demonstrated that the behavioral changes in the OFT were most significantly associated with the expression of inflammation-related genes ( NLRP3, CD163, CD206, F4/80, TMEM119, and TMEM176a), but not glutamate- or GABA-related genes in PLX3397- and MK-801-treated mice. Thus, our results suggest that microglial depletion via a CSF1R/c-Kit kinase inhibitor can ameliorate the hyperactivity induced by an NMDAR antagonist, which is associated with modulation of immune-related genes in the brain.

Dynamic changes in microglia in the mouse hippocampus during administration and withdrawal of the CSF1R inhibitor PLX3397.

Pexidartinib (PLX3397), a colony-stimulating factor-1 receptor (CSF1R) inhibitor, is currently in phase 1-3 clinical trials as a treatment for a variety of tumours. CSF1R signalling regulates the development, survival and maintenance of microglia, the resident brain innate immune cells. In this study, we examined the effects of PLX3397 in the drinking water of mice on microglia in the hippocampus using ionized calcium-binding adapter molecule 1 (Iba1, a microglial marker) immunocytochemistry. A high concentration of PLX3397 (1 mg/mL) significantly decreased the density of Iba1-immunoreactive cells after 7 days of exposure, but a low concentration of PLX3397 (0.5 mg/mL) did not. In addition, both low and high concentrations of PLX3397 significantly increased the intersection number, total length and maximum length of microglial processes in male mice. PLX3397 administered for 21 days eliminated microglia with 78% efficiency in males and 84% efficiency in females. Significant increases in microglial processes were found after both seven and 21 days of PLX3397 exposure in males, whereas decreases in microglial processes were observed after both 14 and 21 days of exposure in females. After PLX3397 withdrawal following its administration for 14 days in males, the soma size quickly returned to normal levels within a week. However, the microglial density, intersection number and total length of microglial processes after 3 days of recovery stabilized to untreated levels. In summary, these findings provide detailed insight into the dynamic changes in microglial number and morphology in the hippocampus in a dose- and time-dependent manner after PLX3397 treatment and withdrawal.

Bis (2-butoxyethyl) Phthalate Delays Puberty Onset by Increasing Oxidative Stress and Apoptosis in Leydig Cells in Rats.

Objective: This study investigated the effects of bis (2-butoxyethyl) phthalate (BBOP) on the onset of male puberty by affecting Leydig cell development in rats. Methods: Thirty 35-day-old male Sprague-Dawley rats were randomly allocated to five groups mg/kg bw per day that were gavaged for 21 days with BBOP at 0, 10, 100, 250, or 500 mg/kg bw per day. The hormone profiles; Leydig cell morphological metrics; mRNA and protein levels; oxidative stress; and AKT, mTOR, ERK1/2, and GSK3ß pathways were assessed. Results: BBOP at 250 and/or 500 mg/kg bw per day decreased serum testosterone, luteinizing hormone, and follicle-stimulating hormone levels mg/kg bw per day (P < 0.05). BBOP at 500 mg/kg bw per day decreased Leydig cell number mg/kg bw per day and downregulated Cyp11a1, Insl3, Hsd11b1, and Dhh in the testes, and Lhb and Fshb mRNAs in the pituitary gland (P < 0.05). The malondialdehyde content in the testis significantly increased, while Sod1 and Sod2 mRNAs were markedly down-regulated, by BBOP treatment at 250-500 mg/kg bw per day (P < 0.05). Furthermore, BBOP at 500 mg/kg bw per day decreased AKT1/AKT2, mTOR, and ERK1/2 phosphorylation, and GSK3ß and SIRT1 levels mg/kg bw per day (P < 0.05). Finally, BBOP at 100 or 500 µmol/L induced ROS and apoptosis in Leydig cells after 24 h of treatment in vitro (P < 0.05). Conclusion: BBOP delays puberty onset by increasing oxidative stress and apoptosis in Leydig cells in rats.The graphical abstract is available on the website www.besjournal.com.

Structure, magnetism and magnetocaloric effect in a new triangular lattice compound Gd3Cu9(OH)19Br8.

A new triangular lattice compound Gd3Cu9(OH)19Br8 has been synthesized by the hydrothermal method. The structure, magnetism and magnetocaloric effect of Gd3Cu9(OH)19Br8 have been studied by X-ray diffraction, magnetic susceptibility, isothermal magnetization and specific heat measurements. In Gd3Cu9(OH)19Br8, the Cu2+ ions form a Kagome lattice along the ab plane, and Gd3+ ions are located in the center of hexagonal holes of the Kagome layer. The Cu-sublattice and Gd-sublattice overlap and constitute a magnetic triangular lattice. The temperature dependence of susceptibility and specific heat curves indicate no magnetic transition down to 2 K, suggesting a paramagnetic-like behavior at low temperature. The magnetocaloric effect (MCE) at low temperature has been calculated according to Maxwell's equations. The maximum value of magnetic entropy change -ΔS M is 26.04 J kg-1 K-1 and adiabatic temperature change ΔT ad is 13.79 K, for a field change of 0-7 T, indicating a potential application of this compound in the field of magnetic refrigeration at low temperature. The influence of 4f-3d interaction on magnetism and MCE is also discussed.

Chronic lithium treatment ameliorates ketamine-induced mania-like behavior via the PI3K-AKT signaling pathway.

Ketamine, a rapid-acting antidepressant drug, has been used to treat major depressive disorder and bipolar disorder (BD). Recent studies have shown that ketamine may increase the potential risk of treatment-induced mania in patients. Ketamine has also been applied to establish animal models of mania. At present, however, the underlying mechanism is still unclear. In the current study, we found that chronic lithium exposure attenuated ketamine-induced mania-like behavior and c-Fos expression in the medial prefrontal cortex (mPFC) of adult male mice. Transcriptome sequencing was performed to determine the effect of lithium administration on the transcriptome of the PFC in ketamine-treated mice, showing inactivation of the phosphoinositide 3-kinase (PI3K)-protein kinase B (AKT) signaling pathway. Pharmacological inhibition of AKT signaling by MK2206 (40 mg/kg), a selective AKT inhibitor, reversed ketamine-induced mania. Furthermore, selective knockdown of AKT via AAV-AKT-shRNA-EGFP in the mPFC also reversed ketamine-induced mania-like behavior. Importantly, pharmacological activation of AKT signaling by SC79 (40 mg/kg), an AKT activator, contributed to mania in low-dose ketamine-treated mice. Inhibition of PI3K signaling by LY294002 (25 mg/kg), a specific PI3K inhibitor, reversed the mania-like behavior in ketamine-treated mice. However, pharmacological inhibition of mammalian target of rapamycin (mTOR) signaling with rapamycin (10 mg/kg), a specific mTOR inhibitor, had no effect on ketamine-induced mania-like behavior. These results suggest that chronic lithium treatment ameliorates ketamine-induced mania-like behavior via the PI3K-AKT signaling pathway, which may be a novel target for the development of BD treatment.

Interlayer quantum transport in Dirac semimetal BaGa2.

The quantum limit is quite easy to achieve once the band crossing exists exactly at the Fermi level (EF) in topological semimetals. In multilayered Dirac fermion systems, the density of Dirac fermions on the zeroth Landau levels (LLs) increases in proportion to the magnetic field, resulting in intriguing angle- and field-dependent interlayer tunneling conductivity near the quantum limit. BaGa2 is an example of a multilayered Dirac semimetal with its quasi-2D Dirac cone located at EF, providing a good platform to study its interlayer transport properties. In this paper, we report the negative interlayer magnetoresistance induced by the tunneling of Dirac fermions between the zeroth LLs of neighboring Ga layers in BaGa2. When the field deviates from the c-axis, the interlayer resistivity ρzz(θ) increases and finally results in a peak with the applied field perpendicular to the c-axis. These unusual interlayer transport properties are observed together in the Dirac semimetal under ambient pressure and are well explained by the model of tunneling between Dirac fermions in the quantum limit.

Psychophysiological Effects of Bamboo Plants on Adults.

The present study was conducted to clarify the psychophysiological relaxation effects of viewing bamboo on university students. Forty healthy Chinese participants enrolled in this study to clarify the psychophysiological relaxation effects of viewing bamboo. The effects of visual stimulation using a pot both with and without a bamboo were recorded by measuring the student's blood pressure, EEG and STAI. We observed that viewing bamboo plants resulted in significantly lower systolic (female, P < 0.001; male, P < 0.001; P < 0.05) and diastolic (female, P < 0.001; male, P < 0.001; P < 0.05) blood pressures, but no changes in the pulse rate (female, P = 0.09; male, P = 0.07; P > 0.05) were observed. The results of the EEG analysis indicated brainwave variation (all P < 0.05) and lower anxiety scores (P < 0.01) after 3 min of viewing bamboo compared with the control. These findings indicate that visual stimulation with bamboo plants induced psychophysiological relaxation effects on adults.

Metabolic Coupling Determines the Activity: Comparison of 11ß-Hydroxysteroid Dehydrogenase 1 and Its Coupling between Liver Parenchymal Cells and Testicular Leydig Cells.

BACKGROUND: 11ß-hydroxysteroid dehydrogenase 1 (11ß-HSD1) interconverts active 11ß-hydroxyl glucocorticoids and inactive 11keto forms. However, its directionality is determined by availability of NADP+/NADPH. In liver cells, 11ß-HSD1 behaves as a primary reductase, while in Leydig cells it acts as a primary oxidase. However, the exact mechanism is not clear. The direction of 11ß-HSD1 has been proposed to be regulated by hexose-6-phosphate dehydrogenase (H6PDH), which catalyzes glucose-6-phosphate (G6P) to generate NADPH that drives 11ß-HSD1 towards reduction. METHODOLOGY: To examine the coupling between 11ß-HSD1 and H6PDH, we added G6P to rat and human liver and testis or Leydig cell microsomes, and 11ß-HSD1 activity was measured by radiometry. RESULTS AND CONCLUSIONS: G6P stimulated 11ß-HSD1 reductase activity in rat (3 fold) or human liver (1.5 fold), but not at all in testis. S3483, a G6P transporter inhibitor, reversed the G6P-mediated increases of 11ß-HSD1 reductase activity. We compared the extent to which 11ß-HSD1 in rat Leydig and liver cells might be coupled to H6PDH. In order to clarify the location of H6PDH within the testis, we used the Leydig cell toxicant ethane dimethanesulfonate (EDS) to selectively deplete Leydig cells. The depletion of Leydig cells eliminated Hsd11b1 (encoding 11ß-HSD1) expression but did not affect the expression of H6pd (encoding H6PDH) and Slc37a4 (encoding G6P transporter). H6pd mRNA level and H6PDH activity were barely detectable in purified rat Leydig cells. In conclusion, the availability of H6PDH determines the different direction of 11ß-HSD1 in liver and Leydig cells.

Simultaneous determination of liensinine, isoliensinine and neferine in rat plasma by UPLC-MS/MS and application of the technique to pharmacokinetic studies.

ETHNOPHARMACOLOGICAL RELEVANCE: The in vivo effects of traditional herbal medicines are generally mediated by multiple bioactive components. The main constituents of Lotus Plumule are alkaloids such as liensinine, isoliensinine and neferine. In this study, a simple, sensitive, and robust analytical method based on ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) has been developed for the determination of the three alkaloids in rat plasma using carbamazepine as internal standard (IS). MATERIALS AND METHODS: After precipitation of the proteins with acetonitrile, chromatography was performed on an Acquity UPLC BEH C18 column (2.1mm×50mm, 1.7µm particle size) using a gradient elution with 0.1% formic acid in water and acetonitrile. Mass spectrometry involved positive electrospray ionization and multiple reaction monitoring (MRM) of the transitions at m/z 611.7→206.2 for liensinine, 611.3→192.2 for isoliensinine, 625.2→206.1 for neferine and m/z 237.1→194.2 for IS. RESULTS: The method was linear over the concentration range 5-1000ng/mL with a lower limit of quantifof 5ng/mL for each alkaloid. Inter- and intra-day precision (RSD%) were all within 11.4% and the accuracy (RE%) were equal or lower than 10.4%. Recoveries were more than 75.3% and matrix effects were not significant. Stability studies showed that the three alkaloids were stable under a variety of storage conditions. CONCLUSION: The method was successfully applied to a pharmacokinetic study involving intravenous administration of liensinine, isoliensinine and neferine to rats.

Changes in BMI before and during economic development and subsequent risk of cardiovascular disease and total mortality: a 35-year follow-up study in China.

OBJECTIVE: It is unclear whether changes in BMI during rapid economic development influence subsequent mortality. RESEARCH DESIGN AND METHODS: We analyzed whether BMI in 1976 and 1994 and changes in BMI during 1976-1994 predict cardiovascular disease (CVD) and all-cause mortality in a 35-year follow-up cohort of 1,696 Chinese (1,124 men and 572 women, aged 35-65 years) in Xi'an, China. Participants were categorized as underweight (<18.5 kg/m(2)), normal weight (18.5-24.9 kg/m(2)), and overweight (≥25.0 kg/m(2)). RESULTS: During 51,611 person-years of follow-up, we identified 655 deaths from all causes and 234 from CVD. From 1976 to 1994, the prevalence of overweight rose from 9.2 to 27.8%. With each unit increment in 1976 BMI, multivariate hazard ratios (HRs) (95% CI) were 0.78 (0.72-0.84) for CVD and 0.91 (0.87-0.95) for all-cause mortality. In contrast, corresponding HRs were 1.14 (1.08-1.19) and 1.05 (1.01-1.08) in 1994 BMI. The HRs for each unit increment in BMI change from 1976 to 1994 were 1.35 (1.25-1.41) for CVD and 1.09 (1.05-1.13) for all-cause mortality. Compared with participants with stable normal weight in 1976 and 1994, HRs of all-cause mortality for those who had normal weight in 1976 but became overweight in 1994 and for those who were persistently overweight during 1976-1994 were 1.42 (1.12-1.80) and 1.80 (1.04-3.14), respectively. CONCLUSIONS: Gaining weight with increased BMI at middle age in Chinese during economic development was associated with elevated risks of all-cause and CVD mortality. Higher BMI measured before economic development was associated with lower mortality risk, whereas BMI measured afterward was associated with increased mortality.

[Relationship between brachial-ankle pulse wave velocity and metabolic syndrome].

OBJECTIVE: To evaluate the association between brachial-ankle pulse wave velocity (ba-PWV) and metabolic syndrome (MetS) among the Chinese elderly and the gender difference. METHODS: We conducted a population-based cross-sectional study in a representative urban area of Beijing, China. A sample of 2 102 community elderly (848 males, and 1 254 females) aged 60 to 95 years were included in the study. MetS was defined according to the 2009 harmonizing definition. RESULTS: The prevalence of MetS was 59.1% (50.1% in males and 65.2% in females, P<0.001). The baPWV value was between 8.2 to 45.6 (20.0 ± 4.4) m/s, and showed an increasing trend with age (P<0.001). The partial correlation showed baPWV was positively associated with BMI (r=0.076, P=0.037), systolic blood pressure (r=0.380, P<0.001), diastolic blood pressure (r=0.276, P<0.001), triglyceride (r=0.040, P=0.046), fasting blood glucose (r=0.140, P<0.001), 2-hour post-meal blood glucose (r=0.121, P<0.001), and negatively associated with HDL-C (r=-0.128, P=0.048). There was a strong association between baPWV and prevalence of MetS and its component number in females but not in males. Compared with the lowest quartile of baPWV, the adjusted ORs were 1.22 (95%CI 0.83-1.79), 1.32 (95%CI 0.90-1.93), 1.46(95%CI 1.00-2.14) in males and 1.28 (95%CI 0.93-1.77), 1.55 (95%CI 1.12-2.16), 1.86(95%CI 1.32-2.61) in females for the second, third and top quartiles. CONCLUSION: The prevalence of MetS increases substantially with increasing levels of baPWV among the Chinese elderly, especially in females.

A real-time PCR for the detection of infectious myonecrosis virus in penaeid shrimp.

Infectious myonecrosis virus (IMNV) is a recently observed shrimp virus, which threats the cultured Litopenaeus vannamei and can cause huge economic loss in shrimp farming industry. The specific aim of this study was to develop a new sensitive real-time PCR method for the specific detection of shrimp IMNV. A real-time PCR assay with a pair of primers to specifically amplify a 101bp IMNV cDNA fragment and a corresponding TaqMan probe was developed, which shown to be specific for IMNV without cross reaction with DNA samples prepared from four other shrimp viruses including white spot syndrome virus (WSSV), hepatopancreatic parvovirus (HPV), monodon baculovirus (MBV), and infectious hypodermal and haematopoietic virus (IHHNV). The method could detect as low as one single copy of IMNV plasmid cDNA.