摘要
ObjectiveTo explore the effect and mechanism of Hei Xiaoyaosan in regulating the tumor necrosis factor receptor superfamily member 6 （Fas）/Fas ligand （FasL）/cysteine protease-8 （Caspase-8）/cysteine protease-3 （Caspase-3） signaling pathway to intervene in neuronal apoptosis and prevent Alzheimer's disease （AD）. MethodNinety SPF-grade SD male rats of 4 months old were selected and randomly grouped as follows： 10 rats in the blank group， 10 rats in the sham group （bilateral hippocampus injected with 1 μL normal saline）， and 70 rats in the modeling group ［bilater hippocampus injected with 1 μL amyloid-beta protein 1-42 （Aβ1-42） solution for the modeling of AD］. Fifty successfully modeled rats were selected and randomly assigned into model， donepezil hydrochloride （0.45 mg·kg-1）， and high-， medium-， and low-dose （15.30， 7.65， 3.82 g·kg-1） Hei Xiaoyaosan groups. Rats were administrated with corresponding agents by gavage once a day for 42 days. Terminal-deoxynucleoitidyl transferase-mediated nick end labeling （TUNEL） was employed to observe the apoptosis of neurons in the cortex and hippocampus， and immunohistochemistry （IHC） was used to detect the expression of B-cell lymphoma-2 （Bcl-2） and Bcl-2-associated X protein （Bax） in the hippocampus. Real-time fluorescence quantitative polymerase chain reaction（Real-time PCR） was employed to determine the mRNA levels of Fas， FasL， and Fas-associated protein with death domain （Fadd）. Western blot was used to determine the protein levels of Fas， FasL， Fadd， Caspase-3， cleved Caspase-3， Caspase-8， and cleved Caspase-8. ResultCompared with the blank group and sham group， the model group showed increased apoptosis rate in the cortex and hippocampus （P<0.01）， elevated Bax level （P<0.01）， lowered Bcl-2 level （P<0.01）， up-regulated mRNA levels of Fas， FasL， and Fadd in the hippocampus （P<0.01）， and up-regulated protein levels of Fas， FasL， Fadd， cleaved Caspase-3， and cleaved Caspase-8 （P<0.01）. Compared with the model group， donepezil hydrochloride and Hei Xiaoyaosan at high and medium doses decreased the apoptosis rate in the cortex and hippocampus （P<0.05， P<0.01）， lowered the Bax level （P<0.01）， elevated the Bcl-2 level （P<0.01）， and down-regulated the mRNA levels of Fas， FasL， and Fadd and the protein levels of Fas， FasL， Fadd， cleaved Caspase-3， and cleaved Caspase-8 （P<0.05， P<0.01） in the hippocampus. Low-dose Hei Xiaoyaosan decreased the apoptosis rate in the cortex and hippocampus （P<0.05， P<0.01）， lowered the Bcl-2 level （P<0.01）， and down-regulated the mRNA levels of FasL and Fadd （P<0.05） and the protein levels of Fas， FasL， Fadd， cleaved Caspase-3， and cleaved Caspase-8 （P<0.05） in the hippocampus. ConclusionHei Xiaoyaosan can protect neurons in the cortex and hippocampus of AD rats by inhibiting the apoptosis mediated by the Fas/FasL/Caspase-8/Caspase-3 signaling pathway.
摘要
ObjectiveTo explore the effect and mechanism of Hei Xiaoyaosan in modulating the synaptic plasticity in APP/PS1 mice by regulating the cyclic adenosine monophosphate （cAMP）/protein kinase A （PKA）/N-methyl-D-aspartate receptor （NMDAR） signaling pathway. MethodTwelve 4-month-old male C57BL/6J mice were selected as the blank control group， and 60 4-month-old male APP/PS1 double transgenic mice were randomized into model， KW-6002 （adenosine receptor antagonist， 3 mg·kg-1）， and high-， medium-， and low-dose （22.10， 11.05， 5.53 g·kg-1， respectively） Hei Xiaoyaosan groups， with 12 mice in each group. Mice were administrated with corresponding drugs for 90 days. Transmission electron microscopy was employed to observe the synaptic ultrastructure of hippocampal neurons， and Golgi staining was used to observe the dendritic spine density of neurons in hippocampal CA1 region. Western blot was employed to measure the protein levels of cAMP， PKA， N-methyl-D-aspartate receptors 1， 2A， and 2B （NR1， NR2A， and NR2B， respectively）， postsynaptic density protein 95 （PSD95）， and synapsin 1 （SYN1）. Real-time fluorescence quantitative polymerase chain reaction（Real-time PCR） was performed to determine the mRNA levels of cAMP， PKA， and NR1. Enzyme-linked immunosorbent assay was employed to determine the content of interleukin-12 （IL-12） and interleukin-4 （IL-4） in the hippocampus. ResultCompared with the blank group， the model group showed blurred boundaries between presynaptic membrane and postsynaptic membrane in hippocampal CA1 region， reduced and scattered synaptic vesicles， and decreased density of postsynaptic membrane， and irregular， disarranged， and loosened dendritic spines of neurons in hippocampal CA1 region （P<0.01）. In addition， the model group presented down-regulated protein levels of cAMP， PKA， NR1， NR2A， NR2B， PSD95， and SYN1 and mRNA levels of cAMP， PKA， and NR1， elevated IL-12 level， and lowered IL-4 level in the hippocampus （P<0.01）. Compared with the model group， the drug intervention groups showed clear and intact boundaries between presynaptic membrane and postsynaptic membrane in hippocampal CA1 region， increased synaptic vesicles with dense arrangement， increased density of postsynaptic membrane， and improved morphology， arrangement， and density of neuronal dendritic spines （P<0.05， P<0.01）. In addition， the drug interventions up-regulated the protein levels of cAMP， PKA， NR1， NR2A， NR2B， PSD95， and SYN1 （P<0.05，P<0.01） and mRNA levels of cAMP， PKA， and NR1 （P<0.01）， lowered the IL-12 level （P<0.01）， and elevated the IL-4 level （P<0.01） in the hippocampus. ConclusionHei Xiaoyaosan can improve the structure and morphology of hippocampal neurons in APP/PS1 mice by activating the cAMP/PKA/NMDAR signaling pathway and repairing synaptic plasticity.
摘要
ObjectiveTo investigate the role and mechanism of Hei Xiaoyaosan in intervening in oxidative stress in the rat model of Alzheimer's disease （AD） via modulating the rat sarcoma （RAS）/rapidly accelerating fibrosarcoma （RAF）/mitogen-activated protein kinase kinase （MEK）/extracellular signal-regulated kinase （ERK） signaling pathway. MethodOne hundred 4-month-old SPF-grade Wistar male rats were randomly grouped as follows： 10 in the blank group， 10 in the sham group （bilateral hippocampus injected with 1 μL normal saline）， and 80 in the modeling group ［bilateral hippocampus injected with 1 μL amyloid beta protein 1-42 （Aβ1-42） solution for the modeling of AD］. Fifty rats qualified for modeling were selected and randomized into the model， donepezil hydrochloride （0.5 mg·kg-1）， and high-， medium-， and low-dose （15.30， 7.65， 3.82 g·kg-1， respectively） Hei Xiaoyaosan groups. The rats were administrated with corresponding drugs by gavage once a day for 42 consecutive days. At the end of gavage， Morris water maze test was performed to examine the learning and memory abilities of the rats， and Nissl staining was used to observe the pathological changes of neurons in CA3 region of the hippocampus. The immunofluorescence assay was used to observe Aβ deposition and tau phosphorylation. Western blot was employed to determine the protein levels of RAS， RAF， phosphorylated （p）-RAF， MEK， p-MEK， ERK， and p-ERK in the hippocampal tissue. Biochemical methods were used to determine the levels of reactive oxygen species （ROS）， malondialdehyde （MDA）， and superoxide dismutase （SOD） in the hippocampal tissue. ResultCompared with the sham group， the model group showed prolonged escape latency （P<0.01）， shortened swimming distance in the target quadrant （P<0.01）， reduced and uneven stained Nissl bodies， enhanced fluorescence intensity of Aβ and p-tau （P<0.01）， up-regulated protein levels of RAS， p-RAF， p-MEK， and p-ERK in the hippocampal tissue （P<0.01）， increased ROS and MDA content （P<0.01）， and decreased SOD activity （P<0.01） on day 5. Compared with the model group， donepezil hydrochloride and high-， medium-， and low-dose Hei Xiaoyaosan shortened the escape latency （P<0.01）， increased the swimming distance in the target quadrant （P<0.01）， improved the arrangement， morphology， and structures of neurons and the number and distribution of Nissl bodies， decreased the fluorescence intensity of Aβ and p-tau （P<0.01）， up-regulated the protein levels of RAS， p-RAF， p-MEK， and p-ERK （P<0.05， P<0.01）， decreased the ROS and MDA content （P<0.01）， and increased the SOD activity （P<0.01） on day 5. ConclusionHei Xiaoyaosan may ameliorate oxidative stress， reduce Aβ and p-tau levels， and inhibit hippocampal neuronal damage by regulating the RAS/RAF/MEK/ERK signaling pathway， thus improving learning and memory abilities.
摘要
Alzheimer's disease （AD） is a neurodegenerative disease that predominantly affects the elderly. It belongs to the category of dementia in traditional Chinese medicine （TCM）， with the onset and progression closely associated with the functions of the kidney， liver， and spleen. The classic TCM formula Hei Xiaoyaosan， which regulates the three Yin of liver， spleen， and kidney， shows broad prospects in treating neurodegenerative diseases. This article reviews the experimental studies reported in the past decade to summarize the mechanisms of Hei Xiaoyaosan and its active components in intervening in AD. Hei Xiaoyaosan can treat AD via multiple targets， levels， and aspects comprehensively. The clinical studies have demonstrated that Hei Xiaoyaosan alone or in combination with other therapies has a definite therapeutic effect on AD. Specifically， it can ameliorate the cognitive impairment， mitigate oxidative stress， and inhibit the overexpression of soluble apoptotic factors in AD patients. This review aims to provide a theoretical basis for the treatment of AD with Hei Xiaoyaosan and explore new research directions. Moreover， it gives new insights into the clinical application of Hei Xiaoyaosan and the development of products with both medicinal and edible values.
摘要
ObjectiveTo investigate the effect of Danzhi Xiaoyaosan on the phosphorylation of tau protein and different sites of glycogen synthase kinase-3β （GSK-3β） and phosphoseryl/suanyl phosphate protein phosphatase 2A （PP2A） in the hippocampus of rats with Alzheimer's disease （AD） and its mechanism. MethodThe rat model of AD was established by injecting okadaic acid into the bilateral hippocampus of 90 male Wistar rats in SPF grades. The rats with successful modeling were selected and randomly divided into model group， aricept group （0.5 mg·kg-1）， and Danzhi Xiaoyaosan high， medium， and low groups （17.55， 8.77， and 4.38 g·kg-1）， and then gavaged for 42 d， once a day. Morris water maze was used to detect the learning and memory ability of rats， Nissl's staining was used to observe the morphological structure of neurons in the hippocampus， and Real-time polymerase chain reaction （Real-time PCR） was used to detect the mRNA expression levels of tau protein， GSK-3β， and PP2A. Western blot was used to determine the protein expression levels of tau protein， GSK-3β， and PP2A. ResultAs compared with the control group， the learning and memory abilities of the rats in the model group were significantly decreased （P<0.01）， and the hippocampal CA3 region cells had abnormal structure， disorderly arrangement， and decreased number. The expression levels of GSK-3β mRNA， GSK-3β， p-GSK-3β-Tyr216， p-PP2A， and p-tau were increased in the model group as compared with the control group （P<0.01）， and those of p-GSK-3β-Ser9 and PP2A decreased significantly （P<0.01）. As compared with the model group， the learning and memory ability of the Aricept group and the Danzhi Xiaoyaosan groups were improved （P<0.05， P<0.01）， and the cell morphology and the number of hippocampal CA3 regions were better. The mRNA expression levels of PP2A and tau in the Aricept group were significantly up-regulated （P<0.05）， the mRNA expression level of GSK-3β was significantly down-regulated （P<0.01）， and the protein expression levels of GSK-3β， p-GSK-3β-Tyr216， and p-PP2A were down-regulated （P<0.05， P<0.01）， and the protein expression level of PP2A was significantly up-regulated （P<0.01）. As compared with the model group， the mRNA expression level of PP2A in the high-dose Danzhi Xiaoyaosan group was significantly up-regulated （P<0.01）， and that of GSK-3β was significantly down-regulated （P<0.01）， whereas the protein expression levels of p-PP2A， p-GSK-3β-Tyr216， and p-tau were down-regulated （P<0.05， P<0.01）， and the protein expression level of PP2A was significantly up-regulated （P<0.01）. As compared with the model group， the mRNA expression level of GSK-3β was significantly down-regulated in the medium-dose Danzhi Xiaoyaosan group （P<0.01）， the protein expression levels of GSK-3β， p-GSK-3β-Tyr216， and p-tau were down-regulated （P<0.05， P<0.01）， and the protein expression level of PP2A was significantly up-regulated （P<0.01）. As compared with the model group， the mRNA expression level of PP2A was significantly up-regulated in the low-dose Danzhi Xiaoyaosan group （P<0.01）， and that of GSK-3β was significantly down-regulated （P<0.01）， whereas the protein expression levels of GSK-3β and p-GSK-3β-Tyr216 were down-regulated （P<0.05， P<0.01）， and those of p-GSK-3β-Ser9 and PP2A were significantly up-regulated （P<0.01）. ConclusionDanzhi Xiaoyaosan can improve the learning and memory ability of rats with AD， and its mechanism may be related to the regulation of the activities of GSK-3β and PP2A protein-related sites and the phosphorylation of tau protein.
摘要
Dermatomyositis is an autoimmune disease involving the skin and muscles. At the onset of dermatomyositis, it is difficult to make an early diagnosis due to atypical clinical manifestations and lack of serological markers. Skin and muscle lesions are associated with disease activity and prognosis in patients with dermatomyositis or clinical amyopathic dermatomyositis. Computed tomography, magnetic resonance imaging, ultrasonography, dermoscopy and other imaging techniques may be used to assess skin and muscle involvements, which can not only improve the accuracy of early diagnosis of dermatomyositis, but also provide important reference for the assessment of disease activity and prognosis.
摘要
Objective@#To evaluate the feasibility and efficacy of naked-eye assessment (NA) of ultrasound-guided fine needle aspiration cytology(US-FNAC) smears, which was performed by a trained non-cytological physician.@*Methods@#A total of 290 smears of FNAC in 143 thyroid nodules were used to evaluate the value of NA by an assistant with more than two years experience of intervention with ultrasound guidance. NA results such as the background of smear (bloody/non-bloody), thickness (thick/thin), as well as the contents (granulated/non-granulated) were recorded. The correlation between NA and cytological results was analyzed. Number of cells under microscopy, the non-diagnostic rate, and the significance between benignity and malignancy with different features of specimens were compared.@*Results@#There was no significant difference between the NA background and cytological findings(P=0.707, 0.200, 0.705, respectively). There was significance in number of cells, non-dianostic rate between thick and thinner specimens (P=0.000, 0.001, respectively). However, there was no significance in benignity and malignancy between thick and thinner specimens (P=0.385). There was significance in cytology results between the granulated and non-granulated smears(all P=0.000). The difference was found in 51.1% of the smears with granulates were malignancy in cytology. While 77.8% of the non-granulated smears were ultimately diagnosed as benignity.@*Conclusions@#The NA could be performed by a trained non-cytological physician and contribute to preliminary predictions of cytology effect.
摘要
ObjectiveTo evaluate the diagnostic value of “taller-than-wide sign” of thyroid malignancy in transverse and longitudinal plane using ultrasound.Methods708 thyroid lesions including 302 benign nodules and 406 malignant nodules ( 390 papillary carcinomas,12 medullary carcinomas,4 follicular carcinomas) that were diagnosed at surgery were included in this study.The sensitivity and specificity of “taller-than-wide sign” in transverse (A/TC ≥ 1 ) and longitudinal plane (A/TL ≥ 1 ) were evaluated using ultrasound.All thyroid malignant lesions were divided into three groups according to size of the nodules:group A≤1 cm; group B≤2 cm,group C＞2 cm.The sensitivity of each group was also evaluated.ResultsThe constituent ratio of “taller-than-wide sign” between benign and malignant nodules was significantly different (P＜0.001 ).The differences of sensitivity of A/TL ≥ 1 and A/TC ≥ 1 among three groups were also found( P ＜0.001),and the sensitivity of A/Tc≥1 was higher than A/TL≥1.ConclusionsA/TC≥1 found by ultrasound contributes to differentiate the thyroid carcinoma from benign nodules.For a large thyroid lesion,other indicators are needed to diagnose thyroid carcinoma.
摘要
OBJECTIVE@#To explore correlation of allergy and outcomes after endoscopic sinus surgery (ESS) in patients with chronic rhinosinusitis.@*METHOD@#Before ESS, 115 cases were arranged to detect allergic serum relative index: the concentration of serum TIgE, specific IgE semi-quantitative test and the concentration of serum ECP; to survey QOL status: VAS, SNOT-20 and SF-36; as well as inquire medical record, Lund-Mackay CT system scoring, clinical classification of chronic rhinosinusitis. After ESS (>1 year), 74 cases were followed up. The subjective and objective assessment were achieved by means of VAS, SNOT-20, SF-36 and Lund-Kennedy endoscopic scores.@*RESULT@#The improvements of VAS scores of the cases with increased serum TIgE were significantly lower than those without increased serum TIgE (P or = 3) were significantly worse than those with positive allergen (<3) (P<0.05). The objective outcomes of the cases with strong positive allergen were significantly worse than those without strong positive allergen (P<0.05). VAS scores and SNOT-20 scores after endoscopic sinus surgery of the cases with strong positive allergen were significantly worse than those without strong positive allergen (P<0.05). The concentration of serum ECP of the invalid cases was significantly higher than effective cases (P<0.05). The concentration of serum ECP was positive correlated with Lund-Kennedy endoscopic scores (r=0.49, P<0.05). In cases of II, III types, the objective outcomes of the cases with increased serum ECP were significantly worse than those without increased serum ECP (P<0.05), VAS scores and SNOT-20 scores after endoscopic sinus surgery of the cases with increased serum ECP were significantly worse than those without incre- ased serum ECP (P<0.05), the improvements of VAS scores of the cases with increased serum ECP were significantly lower than those without increased serum ECP (P<0.05).@*CONCLUSION@#Those allergic serum relative indexes make some negative effect on subjective and objective outcomes. Especially, in cases with polyps, the increase of serum TIgE and serum ECP has more negative effect. Our results might help to choose immunotherapy to combine with endoscopic sinus surgery for those selected patients to improve the outcomes of chronic rhinosinusitis.
Subject(s)
Adolescent , Adult , Aged , Humans , Middle Aged , Young Adult , Allergens , Blood , Cation Transport Proteins , Blood , Chronic Disease , Endoscopy , Eosinophil Cationic Protein , Blood , Immunoglobulin E , Blood , Postoperative Period , Quality of Life , Sinusitis , Blood , General Surgery , Treatment Outcome摘要
OBJECTIVE@#To explore the correlation between allergic factors and chronic rhinosinusitis, and discuss its effects on the development of illness.@*METHOD@#One hundred and one cases were selected to detect allergic serum relative index: the concentration of serum TIgE, specific IgE semi-quantitative test, the concentration of serum ECP as well as inquiring medical record, Lund-Mackay CT system scoring.@*RESULT@#The concentration of serum TIgE and serum ECP in CRS was higher than those in control (P < 0.01). The increased proportion of serum TIgE and serum ECP was significantly different among clinical stages (P < 0.05) and between the cases with and without prior sinus surgery (P < 0.05). CT scores of the cases with serum TIgE and serum ECP increased were significantly higher than those without serum TIgE and serum ECP increased. The strong positive rate in CRS was significantly different among clinical stages (P < 0.05). The strong positive rate increased significantly in the cases with prior sinus surgery (P < 0.05). CT scores in the cases with strong positive allergen were significantly higher than that in the cases without positive allergen (P < 0.05).@*CONCLUSION@#These results suggest that allergic factors make some negative effect on the severity of CRS and are the index for the severity and worse prognosis of illness.