Endothelial KCa3.1 and KCa2.3 Mediate S1P (Sphingosine-1-Phosphate)-Dependent Vasodilation and Blood Pressure Homeostasis.

BACKGROUND: S1P (sphingosine-1-phosphate) has been reported to possess vasodilatory properties, but the underlying pathways are largely unknown. METHODS: Isolated mouse mesenteric artery and endothelial cell models were used to determine S1P-induced vasodilation, intracellular calcium, membrane potentials, and calcium-activated potassium channels (KCa2.3 and KCa3.1 [endothelial small- and intermediate-conductance calcium-activated potassium channels]). Effect of deletion of endothelial S1PR1 (type 1 S1P receptor) on vasodilation and blood pressure was evaluated. RESULTS: Mesenteric arteries subjected to acute S1P stimulation displayed a dose-dependent vasodilation response, which was attenuated by blocking endothelial KCa2.3 or KCa3.1 channels. In cultured human umbilical vein endothelial cells, S1P stimulated immediate membrane potential hyperpolarization following activation of KCa2.3/KCa3.1 with elevated cytosolic Ca2+. Further, chronic S1P stimulation enhanced expression of KCa2.3 and KCa3.1 in human umbilical vein endothelial cells in dose- and time-dependent manners, which was abolished by disrupting either S1PR1-Ca2+ signaling or downstream Ca2+-activated calcineurin/NFAT (nuclear factor of activated T-cells) signaling. By combination of bioinformatics-based binding site prediction and chromatin immunoprecipitation assay, we revealed in human umbilical vein endothelial cells that chronic activation of S1P/S1PR1 promoted NFATc2 nuclear translocation and binding to promoter regions of KCa2.3 and KCa3.1 genes thus to upregulate transcription of these channels. Deletion of endothelial S1PR1 reduced expression of KCa2.3 and KCa3.1 in mesenteric arteries and exacerbated hypertension in mice with angiotensin II infusion. CONCLUSIONS: This study provides evidence for the mechanistic role of KCa2.3/KCa3.1-activated endothelium-dependent hyperpolarization in vasodilation and blood pressure homeostasis in response to S1P. This mechanistic demonstration would facilitate the development of new therapies for cardiovascular diseases associated with hypertension.

Extracellular ATP-induced calcium oscillations regulating the differentiation of osteoblasts through aerobic oxidation metabolism pathways.

INTRODUCTION: The increase of ATP concentration in the extracellular space represents one of the effective signals that stimulate the physiological activities of cells when the bone is exposed to external mechanical stimulation such as stretching and shear stress force throughout life. However, the effects of ATP on osteoblast differentiation and related mechanisms are not well understood. MATERIALS AND METHODS: In this study, the roles of extracellular ATP on osteoblast differentiation, intracellular calcium ([Ca2+]i) levels, metabolomics, and the expression of proteins related to energy metabolism were investigated. RESULTS: Our results showed that 100 µM extracellular ATP initiated intracellular calcium ([Ca2+]i) oscillations via the calcium-sensing receptor (P2R) and promoted the differentiation of MC3T3-E1 cells. Metabolomics analysis showed that the differentiation of MC3T3-E1 cells depended on aerobic oxidation, but little glycolysis. Moreover, the differentiation of MC3T3-E1 cells and aerobic oxidation were suppressed with the inhibition of AMP-activated protein kinase (AMPK). CONCLUSION: These results indicate that calcium oscillations triggered by extracellular ATP can activate aerobic oxidation through AMPK-related signaling pathways and thus promote osteoblast differentiation.

Extracellular Calcium-Induced Calcium Transient Regulating the Proliferation of Osteoblasts through Glycolysis Metabolism Pathways.

During bone remodeling, high extracellular calcium levels accumulated around the resorbing bone tissue as soon as the activation of osteoclasts. However, if and how calcium is involved in the regulation of bone remodeling remains unclear. In this study, the effect of high extracellular calcium concentrations on osteoblast proliferation and differentiation, intracellular calcium ([Ca2+]i) levels, metabolomics, and the expression of proteins related to energy metabolism were investigated. Our results showed that high extracellular calcium levels initiated a [Ca2+]i transient via the calcium-sensing receptor (CaSR) and promoted the proliferation of MC3T3-E1 cells. Metabolomics analysis showed that the proliferation of MC3T3-E1 cells was dependent on aerobic glycolysis, but not the tricarboxylic acid cycle. Moreover, the proliferation and glycolysis of MC3T3-E1 cells were suppressed following the inhibition of AKT. These results indicate that calcium transient triggered by high extracellular calcium levels activated glycolysis via AKT-related signaling pathways and ultimately promoted the proliferation of osteoblasts.

Unsupervised cluster analysis reveals distinct subgroups in healthy population with different exercise responses of cardiorespiratory fitness.

Background: Considerable attention has been paid to interindividual differences in the cardiorespiratory fitness (CRF) response to exercise. However, the complex multifactorial nature of CRF response variability poses a significant challenge to our understanding of this issue. We aimed to explore whether unsupervised clustering can take advantage of large amounts of clinical data and identify latent subgroups with different CRF exercise responses within a healthy population. Methods: 252 healthy participants (99 men, 153 women; 36.8 ± 13.4 yr) completed moderate endurance training on 3 days/week for 4 months, with exercise intensity prescribed based on anaerobic threshold (AT). Detailed clinical measures, including resting vital signs, ECG, cardiorespiratory parameters, echocardiography, heart rate variability, spirometry and laboratory data, were obtained before and after the exercise intervention. Baseline phenotypic variables that were significantly correlated with CRF exercise response were identified and subjected to selection steps, leaving 10 minimally redundant variables, including age, BMI, maximal oxygen uptake (VO2max), maximal heart rate, VO2 at AT as a percentage of VO2max, minute ventilation at AT, interventricular septal thickness of end-systole, E velocity, root mean square of heart rate variability, and hematocrit. Agglomerative hierarchical clustering was performed on these variables to detect latent subgroups that may be associated with different CRF exercise responses. Results: Unsupervised clustering revealed two mutually exclusive groups with distinct baseline phenotypes and CRF exercise responses. The two groups differed markedly in baseline characteristics, initial fitness, echocardiographic measurements, laboratory values, and heart rate variability parameters. A significant improvement in CRF following the 16-week endurance training, expressed by the absolute change in VO2max, was observed only in one of the two groups (3.42 ± 0.4 vs 0.58 ± 0.65 ml⋅kg-1∙min-1, P = 0.002). Assuming a minimal clinically important difference of 3.5 ml⋅kg-1∙min-1 in VO2max, the proportion of population response was 56.1% and 13.9% for group 1 and group 2, respectively (P<0.001). Although group 1 exhibited no significant improvement in CRF at group level, a significant decrease in diastolic blood pressure (70.4 ± 7.8 vs 68.7 ± 7.2 mm Hg, P = 0.027) was observed. Conclusions: Unsupervised learning based on dense phenotypic characteristics identified meaningful subgroups within a healthy population with different CRF responses following standardized aerobic training. Our model could serve as a useful tool for clinicians to develop personalized exercise prescriptions and optimize training effects.

Role of the prefrontal lobe in young normotensives with a family history of hypertension and hypertensives.

Accumulating evidence has demonstrated a significant relationship between prefrontal lobe and hypertension. Elevated blood pressure is usually associated with a prefrontal hemodynamic abnormality. However, the detailed process is still unclear. In this study, we designed a startle protocol and tested the response of the cerebral cortex and cardiovascular system in young normotensive subjects with a family history of hypertension (FH+). Additionally, the cold forehead test (CFT) was performed in hypertensive subjects. In total, 40 young normotensive subjects (21 with FH+ and 19 without a family history of hypertension (FH-)) and 49 middle-aged subjects (21 normotensives (NT) and 28 hypertensives (HT)) were recruited. Our results showed that the magnitude of startle-evoked alpha oscillation at the parasympathetic-related prefrontal cortex (FP1 and FP2) in the FH+ group was significantly smaller than in the FH- group. Acute bradycardia (RRI increase) was observed in FH- subjects but disappeared in the FH+ group. The coupling between instant cardiac acute response (increased RRI) and prefrontal arousal (magnitude of evoked oscillation) was significantly weakened in the FH+ group compared with the FH- group. Furthermore, the decrease in HR induced by parasympathetic outflow during CFT was absent in HT subjects. Hence, we concluded that the impairment of parasympathetic outflow derived from the prefrontal lobe occurs in both healthy young offspring of hypertensive and hypertensive patients.

Increased cardiorespiratory synchronization evoked by a breath controller based on heartbeat detection.

BACKGROUND: The cardiovascular and respiratory systems are functionally related to each other, but the underlying physiologic mechanism of cardiorespiratory coupling (CRC) is unclear. Cardiopulmonary phase synchronization is a form of cardiorespiratory coupling. However, it is difficult to study in experimental data which are very often inherently nonstationary and thus contain only quasiperiodic oscillations. So how to enhance cardiopulmonary synchronization and quantify cardiopulmonary synchronization, the changes in cardiac function under the conditions of cardiopulmonary synchronization, and the physiological mechanisms behind them are the main issues to be discussed in this paper. RESULTS: The results showed that the cardiorespiratory synchronization significantly increased when breathing was controlled by heartbeat detection (p < 0.001). And the respiratory sinus arrhythmia (RSA) obviously decreased (p < 0.01) in the 2/2 mode and increased (p < 0.001) in the 4/4 mode. During the 2/2 breathing pattern compared with spontaneous breathing, systolic blood pressure (SBP) decreased (p < 0.05), and diastolic blood pressure (DBP), mean arterial blood pressure (MBP), and SV decreased significantly (p < 0.01). During the 4/4 breathing pattern compared to 2/2 breathing patterns, DBP, MBP, and cardiac output (CO) increased (p < 0.05), and stroke volume (SV) increased significantly (p < 0.01). When analyzing the relationships among these parameters, the RSA was found to be associated with the respiration rate in all respiratory patterns. CONCLUSIONS: We demonstrated that voluntary cardiorespiratory synchronization (VCRS) can effectively enhance cardiopulmonary phase synchronization, but cardiopulmonary phase synchronization and RSA represent different aspects of the cardiorespiratory interaction. It is found that cardiac function parameters such as the blood pressure and output per stroke could be affected by the number of heartbeats contained in the exhalation and inspiratory phase regulated through VCRS. So we can study cardiopulmonary phase synchronization by VCRS. It can be used to study in experimental data for the physiological mechanism of cardiopulmonary coupling.

The calcium transient characteristics induced by fluid shear stress affect the osteoblast proliferation.

Ca2+ signaling is essential for bone metabolism. Fluid shear stress (FSS), which can induce a rapid release of calcium from endoplasmic reticulum (ER) to produce calcium transients, plays a significant role in osteoblast proliferation and differentiation. However, it is still unclear of how calcium transients induced by FSS activating a number of downstream signals which subsequently regulate cell functions. In this study, we performed a group of Ca2+ transients models, which were induced by FSS to investigate the effects of different magnitudes of Ca2+ transients in osteoblast proliferation. Further, we performed a global proteomic profile of MC3T3-E1 cells in different Ca2+ transients models stimulated by FSS. GO enrichment and KEGG pathway analysis revealed that the TCA cycle was activated in the proliferating process. The activation of TCA needed mitochondrial Ca2+ uptake which were influenced by the amplitude of Ca2+ transients induced by FSS. Our work elucidate that osteoblast proliferation induced by FSS was related to the magnitude of calcium transients, which further activated energetic metabolism signaling pathway. This work revealed further understanding the mechanism of osteoblast proliferation induced by mechanic loading and help us to design new methods for osteoporosis therapy.

Cell micropatterning reveals the modulatory effect of cell shape on proliferation through intracellular calcium transients.

The mechanism by which cell shape regulates the function of the cell is one of the most important biological issues, but it remains unclear. Here, we investigated the effect of the regulation of cell shape on proliferation by using a micropatterning approach to confine MC3T3-E1 cells into specific shapes. Our results show that the proliferation rate for rectangle-, triangle-, square- and circle-shaped osteoblasts increased sequentially and was related to the nuclear shape index (NSI) but not the cell shape index (CSI). Interestingly, intracellular calcium transients also displayed different patterns, with the number of Ca2+ peaks increasing with the NSI in shaped cells. Further causal investigation revealed that the gene expression levels of the inositol 1,4,5-triphosphate receptor 1 (IP3R1) and sarco/endoplasmic reticulum Ca2+-ATPase 2 (SERCA2), two major calcium cycling proteins in the endoplasmic reticulum (ER), were increased with an increase in NSI as a result of nuclear volume changes. Moreover, the down-regulation of IP3R1 and/or SERCA2 using shRNAs in circle-shaped or control osteoblasts resulted in changes in intracellular calcium transient patterns and cell proliferation rates towards that of smaller-NSI-shaped cells. Our results indicate that changes in cell shape changed nuclear morphology and then the gene expression of IP3R1 and SERCA2, which produced different intracellular calcium transient patterns. The patterns of intracellular calcium transients then determined the proliferation rate of the shaped osteoblasts.

[Research progress on chemical constituents and quality control of Tripterygium wilfordii preparations].

Tripterygium wilfordii preparations,with various biological activities such as immunosuppressive,anti-inflammatory and anti-cancer effects,are widely used in the treatment of autoimmune diseases such as rheumatoid arthritis,lupus erythematosus,and nephrotic syndrome. They have definite therapeutic effect,but often cause serious adverse reactions and result in damages to liver,kidney,blood,reproduction,and other systems due to their complex compositions,great toxicity,and narrow margin between the toxic and therapeutic dosages. At present,T. wilfordii preparations produced by different manufacturers exhibit large variations in clinical efficacy and side effects in account of their different chemical compositions and quality fluctuation due to differences in raw materials and production process. However,the existing quality standards are controversial in terms of index components and content limit,which cannot be effectively used for the overall quality control of the preparations. In this paper,the research progress on chemical constituents,quality standard and quality control methods of four T. wilfordii preparations including Tripterygium Tablets,Tripterygium Zongtie Tablets,Tripterygium Shuangceng Tablets and Tripterygium Glycosides Tablets was reviewed,in order to provide ideas and reference for the quality improvement of this type of preparations.

Pulsed electromagnetic fields promote the proliferation and differentiation of osteoblasts by reinforcing intracellular calcium transients.

Pulsed electromagnetic fields (PEMF) can be used to treat bone-related diseases, but the underlying mechanism remains unclear, especially the process by which PEMFs initiate biological effects. In this study, we demonstrated the effects of PEMF on proliferation and differentiation of osteoblasts using the model of calcium transients induced by high extracellular calcium. Our results showed that PEMF can increase both the percentage of responding cells and amplitude of intracellular calcium transients induced by high extracellular calcium stimulation. Compared with corresponding extracellular calcium levels, PEMF stimulation increased proliferation and differentiation of osteoblasts and related gene expressions, such as insulin-like growth factor 1 (IGF-1), alkaline phosphatase (ALP), runt-related transcription factor 2 (Runx2), and osteocalcin (OCN), which can be completely abolished by BAPTA-AM. Moreover, PEMF did not affect proliferation and differentiation of osteoblasts if no intracellular calcium transient was present in osteoblasts during PEMF exposure. Our results revealed that PEMF affects osteoblast proliferation and differentiation through enhanced intracellular calcium transients, which provided a cue to treat bone-related diseases with PEMF. Bioelectromagnetics. 38:541-549, 2017. © 2017 Wiley Periodicals, Inc.

[Reasons for anastomotic leakage following the learning curve by laparoscopic anterior resection of rectal cancer].

OBJECTIVE: To investigate the reasons of anastomotic leakage following learning curve by laparoscopic anterior resection of rectal cancer.
 Methods: From December, 2011 to March, 2015, the clinical information of 179 patients in our hospital who underwent dixon of rectal cancer were collected. The patients were divided into a laparoscopic learning group, a laparotomy group and a laparoscopic group. The reasons of anastomotic leakage for each group were comparatively analyzed. Repeated cutting of anastomotic stoma was compared between the laparoscopic learning group and the laparoscopic group. The male, age, obesity, nutrition complications and the position of anastomotic stoma were compared among the 3 groups.
 Results: The rate of anastomotic leakage in the laparoscopic learning group was significantly higher than that in the laparotomy group and the laparoscopic group (P<0.05). Repeated cutting was a significant risk factor in the laparoscopic learning group (P<0.05), but not in the laparoscopic group. Except obesity, the four factors were significant risk factors in the laparoscopic learning group (P<0.05). All of the five factors were not the significant risk factors in the laparotomy group and the laparoscopic group (P>0.05).
 Conclusion: The operation technical shortcoming is the major factor in the learning of the laparoscopic anterior resection of rectal cancer. In order to reduce the rate of anastomotic leakage in the learning curve period, the selection of patients following the laparoscopic anterior resection of rectal cancer should avoid the following factors: male, older age, the low position of the tumor and the nutrition complications.

Cerebral and neural regulation of cardiovascular activity during mental stress.

BACKGROUND: Mental arithmetic has been verified inducing cerebral and cardiovascular responses. However, the mechanism and sequential responses are still ambiguous. This study aims to reveal the mechanism of cardiovascular and autonomic responses and the related scalp positions that regulate the autonomic nerves system (ANS) during MA task. METHODS: 34 healthy male subjects aged between 19 and 27 years old (mean age 23.6 ± 2.3 years) were recruited in. Electrocardiogram, impedance cardiography, beat-to-beat blood pressure and electroencephalography were measured simultaneously and continuously during the experiments. And the analysis of time-frequency, approximate entropy and Pearson correlation coefficient were adopted. For statistical comparison, paired t test is utilized in the study. RESULTS: The results showed that mental arithmetic task increased heart rate (from 72.35 ± 1.88 to 80.38 ± 2.34), blood pressure (systolic blood pressure: from 112.09 ± 3.23 to 126.79 ± 3.44; diastolic blood pressure: from 74.15 ± 1.93 to 81.20 ± 1.97), and cardiac output (from 8.71 ± 0.30 to 9.68 ± 0.35), and the mental arithmetic induced physiological responses could be divided into two stages, the first stage (10-110 s) and late stage (150-250 s). The high frequency power component (HF) of HRV decreased during MA, but the normalized low frequency power component (nLF) and LF/HF ratio of HRV increased only at the late stage. Moreover, during first stage, the correlations between approximate entropy of electroencephalography at Fp2, Fz, F4, F7 and the corresponding time-frequency results of HF were significant. During the late stage, the correlations between approximate entropy of electroencephalography at Fp2, Fz, C3, C4 and the corresponding nLF was significant. CONCLUSIONS: Our results demonstrated that (1) mental stress induces time-dependent ANS activity and cardiovascular response. (2) Parasympathetic activity is lower during mental arithmetic task, but sympathetic nerve is activated only during late stage of mental arithmetic task. (3) Brain influences the cardiac activity through prefrontal and temporal cortex with the activation of ANS during mental arithmetic.

Effects of extremely low frequency electromagnetic fields on intracellular calcium transients in cardiomyocytes.

Calcium transients play an essential role in cardiomyocytes and electromagnetic fields (EMF) and affect intracellular calcium levels in many types of cells. Effects of EMF on intracellular calcium transients in cardiomyocytes are not well studied. The aim of this study was to assess whether extremely low frequency electromagnetic fields (ELF-EMF) could affect intracellular calcium transients in cardiomyocytes. Cardiomyocytes isolated from neonatal Sprague-Dawley rats were exposed to rectangular-wave pulsed ELF-EMF at four different frequencies (15 Hz, 50 Hz, 75 Hz and 100 Hz) and at a flux density of 2 mT. Intracellular calcium concentration ([Ca(2+)]i) was measured using Fura-2/AM and spectrofluorometry. Perfusion of cardiomyocytes with a high concentration of caffeine (10 mM) was carried out to verify the function of the cardiac Na(+)/Ca(2+) exchanger (NCX) and the activity of sarco(endo)-plasmic reticulum Ca(2+)-ATPase (SERCA2a). The results showed that ELF-EMF enhanced the activities of NCX and SERCA2a, increased [Ca(2+)]i baseline level and frequency of calcium transients in cardiomyocytes and decreased the amplitude of calcium transients and calcium level in sarcoplasmic reticulum. These results indicated that ELF-EMF can regulate calcium-associated activities in cardiomyocytes.

Robustness of cluster synchronous patterns in small-world networks with inter-cluster co-competition balance.

All edges in the classical Watts and Strogatz's small-world network model are unweighted and cooperative (positive). By introducing competitive (negative) inter-cluster edges and assigning edge weights to mimic more realistic networks, this paper develops a modified model which possesses co-competitive weighted couplings and cluster structures while maintaining the common small-world network properties of small average shortest path lengths and large clustering coefficients. Based on theoretical analysis, it is proved that the new model with inter-cluster co-competition balance has an important dynamical property of robust cluster synchronous pattern formation. More precisely, clusters will neither merge nor split regardless of adding or deleting nodes and edges, under the condition of inter-cluster co-competition balance. Numerical simulations demonstrate the robustness of the model against the increase of the coupling strength and several topological variations.

IKZF4/NONO-RAB11FIP3 axis promotes immune evasion in gastric cancer via facilitating PD-L1 endosome recycling.

As an immune checkpoint protein expressed by diverse cancer cells, programmed death ligand 1 (PD-L1) facilitates immune evasion by interacting with programmed cell death-1 (PD-1) on T cells. Despite the clinical benefits observed in various cancer types, strategies targeting PD-1/PD-L1 have demonstrated limited efficacy in gastric cancer (GC). Furthermore, the regulation of PD-L1, especially at post-translational modification levels, remains largely unknown. Therefore, it is crucial to elucidate the mechanisms governing PD-L1 expression to enhance anti-tumor immunity. In this study, we have identified that IKAROS family zinc finger 4 (IKZF4) and Non-POU domain-containing octamer-binding (NONO) synergistically regulate and enhance the expression of RAB11 family-interacting protein 3 (RAB11FIP3) in GC. The IKZF4/NONO-RAB11FIP3 axis facilitates the endosomal recycling of PD-L1, particularly on the cell membrane of GC cells. Moreover, overexpression of RAB11FIP3 mitigates the hypo-expression of PD-L1 protein resulting from IKZF4 or NONO deletion. Functionally, the silencing of RAB11FIP3 or IKZF4 promotes T cell proliferation, and enhances T-cell cytotoxicity towards GC cells in vitro, which further inhibits tumor immune evasion in mice via increasing the infiltration of CD8+ T cells into the tumor microenvironment (TME) to suppress GC progression. Our study suggests that the IKZF4/NONO-RAB11FIP3 axis promotes immune evasion by facilitating PD-L1 endosome recycling, thus presenting a potential therapeutic target for GC treatment.

An oxygen enrichment device for lowlanders ascending to high altitude.

BACKGROUND: When ascending to the high altitude, people living in low altitude areas will suffer from acute mountain sickness. The aim of this study is to test the hypothesis that whether an oxygen concentration membrane can be made and used to construct a new portable oxygen enrichment device for individuals in acute exposure to the high altitude. METHODS: The membrane was fabricated using vinylsiloxane rubber, polyphenylene oxide hydrogen silicone polymers, chloroplatinic acid and isopropyl alcohol. The membrane was assembled in a frame and the performance was tested in terms of concentration of oxygen, flow rate of oxygen enriched air, pressure ratio across the membrane and ambient temperature. Furthermore, the oxygen concentration device was constructed using the membrane, a DC fan, vacuum pump and gas buffer. A nonrandomized preliminary field test was conducted, in which eight healthy male subjects were flown to Tibet (Lhasa, 3,700 m). First, subjects wore the oxygen enrichment device and performed an incremental exercise on cycle ergometer. The test included heart rate (HR), saturation of peripheral oxygen (SpO2) and physical work capacity (PWC). Then, after a rest period of 4 hours, the experimental protocol was repeated without oxygen enrichment device. RESULTS: The testing showed that the membrane could increase the oxygen concentration by up to 30%. Simulation test indicated that although the performance of the oxygen enrichment device decreased with altitudes, the oxygen concentration could still maintain 28% with flow rate of enriched air 110 cm3/s at 5000 m. The field test showed that higher SpO2, lower HR, and better PWC (measured by the PWC-170) were observed from all the subjects using oxygen enrichment device compared with non-using (P < 0.01). CONCLUSIONS: We concluded that the new portable oxygen enrichment device would be effective in improving exercise performance when ascending to the high altitude.

Cluster synchronization induced by one-node clusters in networks with asymmetric negative couplings.

This paper deals with the problem of cluster synchronization in networks with asymmetric negative couplings. By decomposing the coupling matrix into three matrices, and employing Lyapunov function method, sufficient conditions are derived for cluster synchronization. The conditions show that the couplings of multi-node clusters from one-node clusters have beneficial effects on cluster synchronization. Based on the effects of the one-node clusters, an effective and universal control scheme is put forward for the first time. The obtained results may help us better understand the relation between cluster synchronization and cluster structures of the networks. The validity of the control scheme is confirmed through two numerical simulations, in a network with no cluster structure and in a scale-free network.

[Preparation of freeze-dried powder of recombinate hirudin-2 nanoparticle for nasal delivery and permeability through nasal membrane in vitro].

OBJECTIVE: To optimize the freeze-dried powder preparation technology of recombinate hirudin-2 (rHV2) nanoparticle which has bio-adhesive characteristic for nasal delivery, also to investigate its stability and permeability through nasal membrane in vitro. METHODS: Taking the appearance, rediffusion of nanoparticle and rHV2 encapsulation efficiency as the evaluation indexes. Cryoprotector, the preparative technique and the effect of illumination and high temperature factors on its stability for rHV2 freeze-dried powder were investigated. Using Fraze diffusion cell technique, the permeability of rHV2 across rabbit nasal mucous membrane in chitosan solution, chitosan nanoparticle, and nanoparticle frozen-dried powder were compared with that in normal saline solution. RESULTS: The optimized preparation of rHV2 nanoparticle freeze-dried powder was as follows: 5% trehalose and glucose (1:1) was used as cryoprotector, nanoparticle solution was freezed for 24 h in vacuum frozen-dryer after being pre-freezed for 24 h. The content of rHV2 in the freeze-dried powder was 1.1 ug/mg. Illumination had little effect on the appearance, rediffusion and encapsulation efficiency of the rHV2 freeze-dried powder. High temperature could obviously influence the appearance of nanoparticle freeze-dried powder. The permeability coefficient (P) of nanoparticle was 5 times more than that in chictonson solution. It was indicated that chitosan nanoparticle has effect on increasing the permeability of rHV2. CONCLUSION: The freeze-dried powder of chitosan nanoparticle can be a good nasal preparation of rHV2.

Exercise ameliorates anxious behavior and promotes neuroprotection through osteocalcin in VCD-induced menopausal mice.

AIMS: As the ovaries age and women transition to menopause and postmenopause, reduced estradiol levels are associated with anxiety and depression. Exercise contributes to alleviate anxiety and depression and the bone-derived hormone osteocalcin has been reported to be necessary to prevent anxiety-like behaviors. The aim of this study was to investigate the effects of exercise on anxiety behaviors in climacteric mice and whether it was related to osteocalcin. METHODS: Menopausal mouse model was induced by intraperitoneal injection of 4-vinylcyclohexene diepoxide (VCD). Open field, elevated plus maze, and light-dark tests were used to detect anxious behavior in mice. The content of serum osteocalcin was measured and its correlation with anxiety behavior was analyzed. BRDU and NEUN co-localization cells were detected with immunofluorescence. Western blot was applied to obtain apoptosis-related proteins. RESULTS: The VCD mice showed obvious anxiety-like behaviors and 10 weeks of treadmill exercise significantly ameliorated the anxiety and increased circulating osteocalcin in VCD mice. Exercise increased the number of BRDU and NEUN co-localization cells in hippocampal dentate gyrus, reduced the number of impaired hippocampal neurons, inhibited the expression of BAX, cleaved Caspase3, and cleaved PARP, promoted the expression of BCL-2. Importantly, circulating osteocalcin levels were positively associated with the improvements of anxiety, the number of BRDU and NEUN co-localization cells in hippocampal dentate gyrus and negatively related to impaired hippocampal neurons. CONCLUSION: Exercise ameliorates anxiety behavior, promotes hippocampal dentate gyrus neurogenesis, and inhibits hippocampal cell apoptosis in VCD-induced menopausal mice. They are related to circulating osteocalcin, which are increased by exercise.

Investigation of the acicular aragonite growth behavior in AOD stainless steel slag during slurry-phase carbonation.

This study presents a novel method for producing acicular aragonite using argon oxygen decarburization (AOD) slag while controlling the reaction temperature, reaction time, stirring speed, and the magnesium-to­calcium stoichiometric ratio. This approach provides steel plants with an opportunity to decrease their CO2 emissions and promote efficient resource utilization and CO2 storage through the production of high-quality value-added products. The experimental results showed that reaction temperature was the most significant factor affecting the carbonation efficiency of AOD slag, followed by reaction time, stirring speed, CO2 partial pressure, and the liquid-to-solid ratio (L/S). The study also found that elevated temperature and prolonged reaction duration favored the preferential precipitation of aragonite. Additionally, raising the temperature and the magnesium-to­calcium stoichiometric ratio was shown to enhance the formation of aragonite, affecting its crystal growth orientation and dimensions. The optimal combination of reaction parameters for the preparation of acicular aragonite was found to be the reaction time of 8 h, the magnesium-to­calcium stoichiometric ratio of 0.8, the reaction temperature of 120 °C, and the stirring speed of 200 r·min-1. Under these conditions, the resulting acicular aragonite exhibited excellent overall uniformity, a large aspect ratio, and a smooth crystal surface, with a content of 91.49 %, a single crystal length ranging from 9.86 to 32.6 µm, and a diameter ranging from 0.63 to 2.15 µm. This study provides valuable insights into the efficient production of acicular aragonite from steel slag while reducing CO2 emissions and promoting the sustainable use of resources.