ObjectiveTo explore the effect of modified Ditan Decoction （涤痰汤） on chronic intermittent hypoxia cognitive function and the potential function mechanism. MethodsTwenty-four Sprague-Dawley （SD） rats were randomly divided into a normal group， a model group， and a modified Ditan Decoction group， with eight rats in each group. Rats in the modified Ditan Decoction group were administered the decoction by gavage at 14.8 ml/（kg·d）， while the normal group and the model group received the same dose of normal saline. Thirty minutes after daily gavage， the rats in all three groups were placed in an intermittent hypoxia chamber. The oxygen concentration for the model group and the modified Ditan Decoction group was adjusted daily for 8 hours using a computer program to establish the model， while the normal group was exposed to the same airflow rate of ambient air. The intervention was continued for 12 weeks to establish a chronic intermittent hypoxia rat model. The Y-maze test was used to evaluate spatial working memory in the rats. Resting-state functional magnetic resonance imaging （rs-fMRI） was performed to detect whole-brain regional homogeneity （ReHo） and seed-based functional connectivity （FC）. Brain regions showing significant differences in rs-fMRI were selected for further analysis. Immunofluorescence was used to detect β-amyloid （Aβ） deposition and the number of ionized calcium-binding adapter molecule 1 （IBA1）-positive microglial cells. Immunohistochemistry was employed to assess the expression of synaptophysin （SYP）， the excitatory synapse marker vesicular glutamate transporter 1 （Vglut1）， and the inhibitory synapse marker vesicular γ-aminobutyric acid transporter （VGAT）. ResultsCompared with the normal group， the model group showed a reduced spontaneous alternation rate in the Y-maze test. The smoothed Z-score standardized regional homogeneity （SzReHo） value in the left entorhinal cortex significantly increased， and the FC value from this seed point to the left basal forebrain significantly reduced. Additionally， the model group exhibited significantly higher Aβ fluorescence intensity and Iba1 positivity in the left entorhinal cortex， decreased expression of SYP， Vglut1， and VGAT， along with an increased Vglut1/VGAT ratio （P＜0.05 or P＜0.01）. Compared to the model group， the modified Ditan Decoction group demonstrated an increased spontaneous alternation rate， a significantly reduced SzReHo value in the left entorhinal cortex， and a significantly increased FC value from this region to the left basal forebrain. Furthermore， this group showed significantly lower Aβ fluorescence intensity and Iba1 positivity in the left entorhinal cortex， increased levels of SYP， Vglut1， and VGAT， and a decreased Vglut1/VGAT ratio （P＜0.05 or P＜0.01）. ConclusionModified Ditan Decoction can reconstruct the projection from the left basal forebrain to the entorhinal cortex in chronic intermittent hypoxia， thereby reducing Aβ aggregation and excessive microglial activation in the left entorhinal cortex. This process improves the excitation/inhibition imbalance caused by synaptic remodeling， ultimately enhancing cognitive function in rats of chronic intermittent hypoxia.

OBJECTIVES: To explore the neuroprotective mechanism of electroacupuncture at the acupoints Baihui and Shenting in rats with cerebral ischemia-reperfusion (IR) injury. METHODS: Forty-eight male SD rats were equally randomized into sham operation group, cerebral IR model group, acupoint electroacupuncture group and non-acupoint acupuncture group. In the latter 3 groups, cerebral focal ischemic injury was induced using the Longa method; in the two electroacupuncture groups, electroacupuncture was performed either at the acupoints Baihui and Shenting or at non-acupoint sites for 7 days. The changes in neurological deficit scores, cerebral infarction volume, learning and memory function, pathologies in hippocampal CA1 area, neuronal and synaptic ultrastructures, and synaptic density of the rats were observed, and serum GABA level and mRNA and protein expressions of GABA_AR α1, CaMK II, SYN1 and PSD-95 in the hippocampal tissue were detected. RESULTS: Compared with those in cerebral IR model group, the rats receiving electroacupuncture at the acupoints, but not those with electroacupuncture at the non-acupoints, showed significantly decreased neurological deficit scores and cerebral infarction volume with shortened escape latency and increased platform crossings. Electroacupuncture at the acupoints significantly increased neuronal cell number, decreased the width of the synaptic gaps and increased density of synaptic bodies in the ischemic hippocampal CA1 area, resulting also in increased serum GABA levels and hippocampal expressions of GABA_ARα1, SYN1 and PSD-95 and lowered expression level of CaMK II. CONCLUSIONS Electroacupuncture at Baihui and Shenting improves learning and memory function of rats with cerebral IR injury possibly through a mechanism that promotes synaptic regeneration, upregulates hippocampal expressions of GABAAR α 1, SYN1 and PSD-95 and downregulates the expression of CaMK II.
Animals ; Electroacupuncture ; Reperfusion Injury/therapy* ; Male ; Rats ; Rats, Sprague-Dawley ; Brain Ischemia/therapy* ; Hippocampus/metabolism* ; Memory ; Learning ; Disks Large Homolog 4 Protein/metabolism* ; Synapses ; Acupuncture Points ; Calcium-Calmodulin-Dependent Protein Kinase Type 2/metabolism* ; Receptors, GABA-A/metabolism* ; gamma-Aminobutyric Acid/metabolism*

Objective To investigate the relationship between learning and memory deficit and demyelination of the corpus callosum in twelve-month old APP/PS1 transgenic mice. Methods Twelve twelve-month old APP/PS1 transgenic mice were as AD group, and age-matched wild type (WT) littermates were as WT group. Learning and memory ability was tested with Morris water maze, and the mor-phology of nerve fiber of corpus callosum was detected with Luxol Fast Blue staining. Immunohistochemistry was used to detect myelin ba-sic protein (MBP) in the corpus callosum. Thioflavine S staining was used to detect amyloid plaque in the corpus callosum. Results Com-pared with WT group, the latency increased (Z>2.873, P<0.01) and the times crossing the location of the platform decreased (t=-7.339, P<0.001) in AD group. The nerve fibers were sparse and disorganized, with a lot of vacuoles in the corpus callosum of AD group. The positive expression of MBP in the corpus callosum was significantly decreased (t=-4.481, P<0.001) in AD group compared with WT group. There were amyloid plaques in the corpus callosum of AD group. Conclusion Twelve-month old APP/PS1 transgenic mice exhibit learning and memory deficit, which may be attributed to the deposition of the amyloid plaque mediated demyelinated injury of the corpus callosum.

Objective To explore the effects of electroacupuncture at Shenting (DU24) and Baihui (DU20) on cognitive dysfunction after stroke. Methods Forty-five Sprague-Dawley rats were randomly divided into control group (n=15), model group (n=15) and electroacupuncture group (n=15). The latter two groups were occluded their middle cerebral artery for two hours and reperfused. The electroacupuncture group accepted electroacupuncture at Shenting and Baihui 24 hours after modeling for seven days. They were assessed with Morris water maze once a day since the second day of intervention. Their brains were stained with TTC staining to measure cerebral infarction volume after treatment, while the expression of cyclic AMP response element binding protein (CREB) and phosphorylation (p-CREB) in hippocampal CA1 area were detected with immunohistochemistry. Results The escape latency and swimming distance of place navigation shortened in the electroacupuncture group compared with those in the model group (P<0.05) from the fourth day of intervention. The number of cross platform of spatial probe increased (P<0.05). The infarction volume was less in the electroacupuncture group than in the model group (P< 0.05), with increased expression of CREB and p-CREB in hippocampal CA1 area (P<0.05). Conclusion Electroacupuncture at Shenting and Baihui can increase the expression of CREB and phosphorylation in hippocampal CA1 area in rats after cerebral ischemia-reperfusion, to protect the neurons from ischemia and improve the learning and memory function.

BACKGROUND:Mineralized nodules are the mature marker of osteoblast differentiation, and the observation methods mainly use alizarin red staining.
OBJECTIVE:To compare the observation results of mineralized nodules by three methods, and to explore their characteristics and advantages, as wel as further application in the research of bone disease.
METHODS:The rat osteoblast-like cellline UMR-106 were cultured in the fresh medium that was changed every day, for 14 days. Alizarin red staining-light microscope, tetracycline fluorescence labeling-laser confocal scanning microscopy, and scanning electron microscopy were used to observe mineralized nodules. The calcium content of mineralized nodules was quantified using scanning electron microscopy and energy dispersive spectroscopy. In addition, tumor necrosis factor alpha that could inhibit the proliferation and differentiation of osteoblasts was used as the control.
RESULTS AND CONCLUSION:The three morphology methods could be used to observe the mineralized nodules of normal osteoblasts. As for tumor necrosis factor alpha, no mineralized nodules of osteoblasts were observed by alizarin red staining-light microscopy;smal mineralized nodules were observed by tetracycline staining-laser scanning confocal microscopy and scanning electron microscopy, suggesting tetracycline staining and scanning electron microscopy were more sensitive in the observation. Scanning electron microscopy could be used to observe the submicroscopic structures of mineralized nodules in the osteoblasts, and the formation of mineralized nodules, including the calcium secretion. Additional y, scanning electron microscopy combined with energy dispersive spectroscopy analysis can successful y quantify and position the mineralized nodules, indicating a potential application in the research of bone diseases.

BACKGROUND:Tougu Xiaotong capsule is the clinical prescription for the treatment of osteoarthritis in Fujian University of Traditional Chinese Medicine, and the previous studies mainly focus on effect to cartilage.
OBJECTIVE:To observe the effect of Tougu Xiaotong capsule on the proliferation and differentiation of osteoblasts as wel as the expressions of bone remodeling correlated factors.
METHODS:Rat osteoblast-like cel line ROS17/2.8 cel s were incubated with Tougu Xiaotong capsule. The ROS17/2.8 cel s were divided into blank control group and Tougu Xiaotong capsule groups with different
concentrations. The cel proliferation was determined by methylthiazolyldiphenyl-tetrazolium bromide assay. Osteoblast differentiation biomarkers alkaline phosphatase activity, osteocalcin and bone mineralized nodules were measured with colorimetry, enzyme-linked immunosorbent assay and alizarin red staining, respectively. The real-time PCR and enzyme-linked immunosorbent assay were used to detect the expressions of bone remodeling factors osteoprotegerin/receptor activator of nuclear factorκB ligand.
RESULTS AND CONCLUSION:Compared with the control group, the Tougu Xiaotong capsule with the
concentration of 0.25-2 g/L could significantly promote the ROS17/2.8 cel proliferation (P<0.05), up-regulate the alkaline phosphatase activity, osteocalcin expression level and mineralized nodules area, and increase the
percentage of bone remodeling factors osteoprotegerin/receptor activator of nuclear factorκB ligand (P<0.05). The mechanism of Tougu Xiaotong capsule protecting osteoarthritis may partly result from the regulation of
proliferation and differentiation of osteoblasts and bone remodeling.

Microscopic and histochemical methods were used to investigate flavonoids localization in the leaf and the stem of the Sarcandra glabra. The results indicated that flavonoids distributed mainly in epidermis, collenchyma, vascular bundles, secretory cells and palisade tissue of leaf. In the stem, they distributed mainly in epidermis, collenchyma, phloem and secretory cells. Histochemical localization of flavonoids using 5% solution of NaOH is convenient, rapid and reliable. The content of flavonoids in the leaf was higher those than in the stem. For sustainable utilization of the resources we suggested that only the leaves could be harvested.
Flavonoids ; metabolism ; Magnoliopsida ; metabolism ; Microscopy ; Plant Leaves ; metabolism ; Plant Stems ; metabolism