ABSTRACT
Objective:To observe and compare the protective effects of Tongqiao Huoxue decoction （TQHX） prepared by three methods against cerebral ischemia-reperfusion injury （CIRI）， and to explore its mechanism through the glutamate （Glu） metabolic pathway in astrocytes. Method:The male SD rats of SPF grade were subjected to CIRI model induction by the modified middle cerebral artery occlusion method. The model rats were randomly divided into a model group， a sham operation group， and water-decocted， wine-decocted， and alcohol-extracted TQHX （6.3 g·kg<sup>-1</sup>·d<sup>-1</sup>） groups. The rats were treated correspondingly for 7 days. Those in the sham operation group and the model group were treated with an equal volume of normal saline by gavage. After the final treatment， the neurological function of rats was assessed by the modified neurological severity score （mNSS）. Hematoxylin-eosin （HE） staining was used to observe the morphological changes of ischemic brain tissues in rats. High-performance liquid chromatography （HPLC） was used to detect glutamate （Glu） in ischemic brain tissues. The expression of glutamate transporter-1 （GLT-1） and glial fibrillary acidic protein （GFAP） and co-expression of glutamine synthetase （GS） and GFAP in ischemic brain tissues were detected by immunofluorescence assay. Western blot was used to detect the protein expression of GFAP， GLT-1， and GS. Result:Compared with the sham operation group， the model group showed increased mNSS （<italic>P</italic><0.01）， large necrosis of cerebral cortex in ischemic brain tissues with disordered cell arrangement， obscure boundary， intracellular edema， and inflammatory infiltration， elevated Glu in ischemic brain tissues （<italic>P</italic><0.01）， declining GLT-1-GFAP co-expression and GS-GFAP co-expression （<italic>P</italic><0.01）， up-regulated expression of GFAP protein， and reduced protein expression of GLT-1 and GS（<italic>P<</italic>0.05，<italic>P<</italic>0.01）. Compared with the model group， the TQHX groups showed decreased mNSS （<italic>P<</italic>0.01）， relieved injury in the cerebral cortex and hippocampal nerve cells in ischemic brain tissues， reduced Glu expression（<italic>P<</italic>0.05，<italic>P<</italic>0.01）， elevated co-expression of GLT-1 and GFAP （<italic>P<</italic>0.05，<italic>P<</italic>0.01）， and up-regulated protein expression of GFAP and GLT-1（<italic>P<</italic>0.05，<italic>P<</italic>0.01）. The co-expression of GS and GFAP （<italic>P<</italic>0.05，<italic>P<</italic>0.01）and the expression of GS （<italic>P<</italic>0.01）were increased in the wine-decocted and alcohol-extracted TQHX groups. Compared with the water-decocted TQHX group， the alcohol-extracted group showed increased GLT-1-GFAP and GS-GFAP co-expression（<italic>P<</italic>0.05）； the wine-decocted and alcohol-extracted TQHX groups exhibited elevated GS protein expression （<italic>P<</italic>0.05）； the alcohol-extracted TQHX group displayed declining Glu content （<italic>P</italic><0.01） and increased protein expression of GFAP and GLT-1 （<italic>P<</italic>0.05， <italic>P<</italic>0.01）. Compared with the wine-decocted TQHX group， the alcohol-extracted TQHX group showed increased protein expression of GFAP and GLT-1（<italic>P<</italic>0.05，<italic>P<</italic>0.01）. Conclusion:TQHX prepared by three methods can improve neurological deficits in CIRI rats. The effect is presumedly achieved by promoting the further activation of astrocytes， increasing the expression of GLT-1 and GS， promoting the clearance of Glu accumulated in the synaptic cleft by astrocytes through the Glu-glutamine （Gln） circulation， and reducing the excitotoxicity of Glu. The alcohol-extracted TQHX group was superior to the water-decocted and wine-decocted TQHX groups in reducing the content of Glu in ischemic brain tissues， promoting the activation of astrocytes， and enhancing the protein expression of GLT-1 and GS.
ABSTRACT
Objective:To study the protective effect and mechanism of Chaenomelis Fructus alcohol extract (CFE) on the synovium of rheumatoid arthritis (RA). Method:Sixty male SD rats were randomly divided into normal group and model group. RA model was made by injection of complete Freund's adjuvant， and then was randomly divided into model group， CFE low， medium and high dose group and Tripterygium glycoside group according to the inflammatory score. The CFE groups (0.15，0.30，0.60 g·kg-1·d-1) had intragastric administration once a day for 30 d after the model establishment. The blank control group and the model group were given the same volume saline water by gavage. After all the drugs were given， the blood， joint tissues and synovium tissue of rats were collected. The levels of interleukin-1β (IL-1β)， interleukin-6 (IL-6)， interleukin-10 (IL-10) and tumor necrosis factor (TNF)-α in serum were detected by enzyme linked immunosorbent assay (ELISA)， the pathological changes of synovium were observed by hematoxylin eosin (HE) staining， and the expressions of B-cell lymphoma-2 (Bcl-2)，Bcl-2 associated X protein (Bax) and death factor (Fas) in joints were detected by Western blot. Result:Compared with normal group， the swelling degree and inflammation index of rats' feet in model group increased significantly， the levels of inflammatory factors IL-1β， IL-6， TNF-α in serum increased (P<0.01)， anti-inflammatory factor IL-10 decreased (P<0.01)， the protein expression of Bax， Fas and Bcl-2 increased， and the statistical results of Bcl-2 showed significant difference (P<0.05). Compared with model group， the swelling degree and inflammatory index of the plantar of RA rats were improved in the middle and high dose groups of CFE (P<0.01)， the pathological changes such as synovial tissue hyperplasia and inflammatory cell infiltration were reduced in each dose group， and the levels of IL-1β， IL-6 and TNF-α in serum were reduced (P<0.05)， anti inflammatory factor IL-10 increased (P<0.05)， the disorder of inflammatory cytokine in the model was corrected， Bax， Fas expression increased， Bcl-2 protein expression decreased (P<0.01). Conclusion:CFE can reduce the degree of inflammation in RA joint and has obvious anti RA effects， which may be related to the apoptosis of synoviocytes induced by CFE.
ABSTRACT
Supernumerary teeth is one of the dysplasia that the number of the teeth are more than physical number. Most cases of reports were with 1-2 supernumerary teeth and rare cases were with more than 3 supernumerary teeth. A 17-year old female patient of 7 impacted supernumerary teeth were found because of toothache of premolar caused by impacted supernumerary teeth and were treated by extraction of impacted supernumerary teeth.
Subject(s)
Female , Humans , Bicuspid , Incisor , Tooth, Impacted , Tooth, SupernumeraryABSTRACT
Incidence rate of 4 root canals in maxillary second molar is very low and most molars have only two mesiobuccal root canals. The emergence of 4 root canals in maxillary second molar with two lingual root canals is especially rare. A case of 4 root canals maxillary second molar with two lingual root canals was successfully treated and reported in this article.
Subject(s)
Humans , Dental Pulp Cavity , Maxilla , Molar , Tongue , Tooth RootABSTRACT
OBJECTIVE@#To observe the inductive efficiency of deriving hematopoietic cells from human embryonic stem (hES) cells co-cultured with human yolk sac stromal cells, fetal liver stromal cells or fetal bone marrow stromal cells,in order to discuss the effect of the different hemopoietic microenvironment on hemopoietic cytogenesis.@*METHODS@#We used two-step method to induce the hES cells into the hematopoietic cells. In the first step the hES cells were co-cultured with cytokines by formation of the day 5 embryoid bodies (5d EBs). In the second step the 5d EB cells were induced into the hematopoietic cells by co-culturing with human yolk sac stromal cells, fetal liver stromal cells or fetal bone marrow stromal cells for 10 days. The inductive efficiencies of deriving hematopoietic cells from hES cells co-cultured with the different hemopoietic microenvironment were reflected by the expression levels of flk, CD34 and CD45 antigen.@*RESULTS@#Flow cytometry analysis demonstrated that the population of the cells co-cultured with human yolk sac stromal cells contained flk (1.80%+/-0.56%), CD34 (1.30%+/-0.14%) or CD45 (1.05%+/-0.63%) positive cells; the population of the cells co-cultured with human fetal liver stromal cells contained flk (34.00%+/-25.45%), CD34 (38.40%+/-24.80%) or CD45 (72.60%+/-25.70%) positive cells; the population of the cells co-cultured with human fetal bone marrow stromal cells contained flk (2.50%+/-1.48%), CD34 (3.20%+/-0.56%) or CD45 (1.65%+/-0.21%) positive cells. Compared with spontaneous differentiation of EBs, all of the three stromal cells could induce EBs into the hematopoietic cells (P<0.05).@*CONCLUSION@#The inductive efficiency of deriving hematopoietic cells from EBs co-cultured with human fetal liver stromal cells was higher than EBs co-cultured with human yolk sac stromal cells and fetal bone marrow stromal cells.