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ObjectiveTo investigate the effect and mechanism of modified Shuyuwan （SYW） on hippocampal myelin sheath injury in vascular dementia （VD） model rats. MethodSixty male SD rats of SPF grade were randomly divided into sham operation group， model group， and high-， medium- and low-dose modified SYW groups， with 12 rats in each group. The VD model was induced by bilateral carotid artery ligation in rats of the groups except for those of the sham operation group. After modeling， rats were screened by the water maze test， followed by drug treatment by gavage. Specifically， rats in the modified SYW groups were treated with modified SYW at 10， 5， 2.5 g·kg-1·d-1， accordingly， and those in other groups were administered with the same amount of normal saline. After intragastric administration for 28 days， the spatial learning and memory abilities of rats were detected by the water maze test. The hippocampal neuron structure was observed by hematoxylin-eosin （HE） staining. The content of hippocampal tumor necrosis factor （TNF）-α， interleukin-6 （IL-6）， and glutamate （Glu） was observed by biochemical detection. The hippocampal expression of myelin basic protein （MBP）， astrocyte activation marker glial fibrillary acidic protein （GFAP）， and connexin 43 （Cx43） was detected by immunofluorescence detection. The myelin sheath structure in the hippocampus was observed by the electron microscope. The α-amino-3-hydroxy-5-methyl-4-isoxazole-propionic acid receptor （AMPAR） and Cx43 protein expression was detected by Western blot. ResultCompared with the sham operation group， the model group showed prolonged escape latency （P<0.01）， decreased times of crossing the original platform and percentage of target quadrant detention time （P<0.01）， disordered neuron structure in the hippocampal CA1 region， loose myelin sheath lamella with blurry edge， up-regulated expression levels of TNF-α， IL-6， and Glu in the hippocampal CA1 region， especially Glu （P<0.01）， reduced expression of AMPAR （P<0.01）， increased protein expression of p-AMPAR and Cx43 （P<0.01）， significantly dwindled protein expression of MBP in the myelin sheath， and enhanced fluorescence co-labeled by GFAP and Cx43. Compared with the model group， the modified SYW groups showed shortened escape latency （P<0.05）， increased times of crossing the original platform and percentage of target quadrant detention time （P<0.05）， closely arranged hippocampal neuron structure， denser myelin sheath lamella with clear edge， down-regulated expression levels of TNF-α， IL-6， and Glu in the hippocampal CA1 region， especially Glu （P<0.01）， up-regulated AMPAR （P<0.01）， reduced protein expression of p-AMPAR and Cx43， especially in the high-dose group （P<0.01）， significantly elevated protein expression of MBP in the myelin sheath， and weakened fluorescence co-labeled by GFAP and Cx43， especially in the high-dose group. ConclusionModified SYW can improve the learning and memory abilities of VD rats， and the mechanism may be related to the inhibition of Cx43 expression， reduction of the release of Glu， inhibition of AMPAR-mediated inflammatory response to reduce the production of astrocyte marker GFAP， and promotion of the expression of MBP protein to alleviate myelin injury.
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OBJECTIVE To compare the clinical characteristics between eosinophilic and noneosinophilic CRSwNP and identify the predictors of eosinophilic CRSwNP. The impact of comorbidity (allergic rhinitis-AR, bronchial asthma-BA) on the predictors was also studied. METHODS Histologic characteristics of surgical samples were analyzed in 182 CRSwNP patients, who were classified into eosinophilic CRSwNP (ECRSwNP) group and noneosinophilic CRSwNP (non-ECRSwNP) group. Factors associated with ECRSwNP were selected by correlation analyses. The optimal cutoff points of the predictors were determined by a receiver operating characteristic curve. Based upon the different comorbidity, four groups were divided to study the impact of comorbidity on the optimal cutoff points of the predictors. RESULTS Patients with ECRSwNP had significant higher rate of revision FESS, significant higher concurrent rate of allergic rhinitis and bronchial asthma than non-ECRSwNP patients (all P<0.05). There were significant differences in Ratio of Lund-Mackay scores, peripheral blood eosinophil percentage and serum total IgE between ECRSwNP group and non-ECRSwNP group except the peripheral blood neutrophilic percentage (U value were 1028.00, 1143.50, 800.00 respectively, all P<0.05). Peripheral blood eosinophilic percentage had the highest significant correlation with degree of infiltration of eosinophils in nasal polyps (r =0.538, 0.568, both P<0.05) and there also existed significant correlation between degree of infiltration of eosinophils in nasal polyps and Lun-Mackay score (r =0.419, P<0.05). Comorbidity of AR and BA could impact the peripheral blood eosinophil percentage. According to the different combination of comorbidity, the range of the optimal cutoff points of peripheral blood eosinophil percentage was 3.05%-4.8%. CONCLUSION ECRSwNP and non-ECRSwNP displayed significant differences in certain clinical features. Comorbidity of AR and BA could increase the peripheral blood eosinophil percentage, that may impact the diagnosis of the subtype of ECRSwNP.
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Hepatocellular carcinoma is a common malignant tumor, which has a poor prognosis and a high mortality rate. Under the influence of respiratory motion, the position error of liver cancer is relatively high. The image-guided radiotherapy is a combination of radiotherapy and imaging device, which can detect and correct the error caused by the movement of the organs and the position of the pendulum to improve the accuracy of radiotherapy.
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Objective To study the methods of matching kilovolt CBCT image with planning CT scan. Methods A total of 121 CBCT scans were matched with planning CT scan using a manual and four automatic match methods by four observers in the offline. In the manual match, the live contour was used as a surrogate for image registration. Four automatic match methods, including routine soft?tissue match, routine bone match, automatic liver match and vertebral body match, were performed using image registration sofeware. First, the stability of the sofeware was tested. Then, the reproducibility of the same automatic match method was evaluated by comparing different observers’ match results. After the manual match by four observers, the mean of the match results was used as a standard to compare with others. The differerces was test by McNemar method. Results In the uniform match factors, automatic match result would not change. The reproducibility of routine soft?tissue and bone match are best, automatic vertebral body match is better than automatic liver match. Howerver, the automatic liver match result is the most similar to manual match, the percentages of match result have an absolute error no more than 3 mm in left?right, superior?inferior and anterior?posterior directions are 84?? 3%, 77?? 7% and 92?? 6%. Conclusions Automatic liver match can be used in image?guided radiotherapy for liver cancer, however, it should be performed by experienced oncologist and technologist together in each fraction, after that, the matching result should be adjusted carefully according to live contour.
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OBJECTIVE@#To explore the expression of interleukin-25 (IL-25) in chronic rhinosinusitis with nasal polyps (CRSwNP) and its potential significance in pathogenesis.@*METHOD@#IL-25 expression in blood was detected by enzyme-linked immunosorbent assay (ELISA). IL-25 expression in tissue was detected by immunohistochemistry (LSAB method) from polyps (68 CRSwNP patients) and 55 inferior turbinate mucosa from patients with deviation of nasal septum served as control. Complete blood count and HE staining of blood and tissue eosinophil infiltration degree.@*RESULT@#IL-25 expression in CRSwNP group were significantly higher than the control group, the difference was statistically significant (P < 0.01). IL-25 expression in local organizations was positively correlated with the number of eosinophil infiltration in CRSwNP group (r = 0.679, P < 0.01).@*CONCLUSION@#The expression of IL-25 in CRSwNP patients mutually reinforcing and might increase eosinophil infiltration and play an important role in the development of CRSwNP.
Subject(s)
Humans , Chronic Disease , Enzyme-Linked Immunosorbent Assay , Interleukin-17 , Blood , Nasal Polyps , Blood , Nasal Septum , Rhinitis , Blood , Sinusitis , Blood , TurbinatesABSTRACT
Chronic rhinosinusitis (CRS) can be controlled by a combination of conservative treatment and surgical procedures. However, there is one group of CRS endotypes, refractory CRS, such as eosinophilic chronic rhinosinusitis with nasal polyp (ECRSwNP), for which the current treatment strategies of anti-inflammatory and/or antibiotic therapy or surgical removal of the lesion to improve sinus drainage are less effective. This lack of treatment efficacy highlights the need for further fundamental and clinical research to improve or restore nasal mucosal function of CRS. The role of the immune response in the pathogenesis of mucosal remodeling, which is a key problem in refractory CRS, is of particular current interest. ECRSwNP characterized by Th17/Treg imbalance in- duced by IL-6 has a different mucosal remodeling pattern. This review intends to illustrate the role of the imbalance between Th17 (T helper 17) and Treg (regulatory T) cells in the mucosal remodeling of ECRSwNP and to suggest a novel therapeutic target on treating ECRSwNP.
Subject(s)
Humans , Chronic Disease , Nasal Mucosa , Nasal Polyps , Metabolism , Rhinitis , Metabolism , Sinusitis , Metabolism , T-Lymphocytes, Regulatory , Th17 CellsABSTRACT
OBJECTIVE@#To study the expression of PDCD4 and its relationship with cell proliferation and apoptosis in laryngeal squamous cell carcinomas and to evaluate its clinical significance.@*METHOD@#The immunohistochemical staining method was applied to investigate the expression of PDCD4 and Ki-67 in 60 laryngeal squamous cell carcinomas tissues and 21 their surrounding laryngeal normal mucosa tissues. TUNEL method was applied to detect the apoptosis. The proliferation index (PI) and apoptosis index (AI) were defined.@*RESULT@#The positive rate of PDCD4 protein was significantly lower in laryngeal squamous cell carcinomas than that in the laryngeal normal mucosa tissues (P < 0.01). The expression of PDCD4 had relation with the differentiation grading and lymph node metastasis (P < 0.05), but not related to tumor sites and TNM stage. The PI of laryngeal squamous cell carcinomas tissues was higher than that of the normal laryngeal mucosa tissues (P < 0.01). No relation was found among PI and tumor sites, differentiation grading and lymph node metastasis. PI had correlation with TNM stage (P < 0.05). The AI of the cancer was higher than that of normal laryngeal mucosa tissues (P < 0.01). There was no relationship between AI and tumor sites, TNM stage and lymph node metastasis. AI correlated with differentiation grading (P < 0.01). The PI in PDCD4 positive group was significantly lower than in PDCD4 negative group (P < 0.05). The AI in PDCD4 positive group was significantly higher than in PDCD4 negative group (P < 0.05).@*CONCLUSION@#The expression of PDCD4 is low in laryngeal squamous cell carcinomas and it may play important roles in occurrence and development of laryngeal squamous cell carcinomas through the inducing apoptosis and inhibiting proliferation.
Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Apoptosis , Apoptosis Regulatory Proteins , Metabolism , Carcinoma, Squamous Cell , Metabolism , Pathology , Cell Proliferation , Laryngeal Mucosa , Metabolism , Pathology , Laryngeal Neoplasms , Metabolism , Pathology , RNA-Binding Proteins , MetabolismABSTRACT
OBJECTIVE@#To study whether programmed cell death 4 (PDCD4) is involved in the development and progression of laryngeal squamous cell carcinomas (LSCC).@*METHOD@#The expressions of PDCD4 protein were examined by immunohistochemical staining and Western blot in LSCC tissues and their surrounding laryngeal normal mucosa tissues.@*RESULT@#The positive rate of PDCD4 protein in LSCC was significantly lower than that in the laryngeal normal mucosa tissues by immunohistochemical staining (P<0.01). No correlation was found between positive expression of PDCD4 and gender, age, tumor site, TNM stage, but there was significant correlation between positive expression of PDCD4 from differentiation grading and lymphatic matastasis. Meanwhile,we got the similar results by Western blot (P<0.01).@*CONCLUSION@#The low expression of PDCD4 protein is closely related to the development and progression of LSCC. PDCD4 might be a novel molecular marker for prognosticating metastases and prognosis of LSCC.
Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Apoptosis Regulatory Proteins , Metabolism , Carcinoma, Squamous Cell , Metabolism , Pathology , Immunohistochemistry , Laryngeal Neoplasms , Metabolism , Pathology , Prognosis , RNA-Binding Proteins , MetabolismABSTRACT
OBJECTIVE@#To study the effect on the expression of protein kinase CK2alpha and growth of human laryngeal carcinoma xenograft in nude mice by applying small interfering RNA (siRNA) specific to protein kinase CK2alpha.@*METHOD@#Human laryngeal carcinoma Hep-2 cells were implanted under the skin of nude mice. After the tumors grew to a definite size, the tumors were injected with siRNA expression plasmid specific to protein kinase CK2alpha. The weight and volume of subcutaneous tumors were measured. The expression level of protein kinase CK2alpha mRNA and protein of tumors were measured with reverse transcription polymerase chain reaction (RT-PCR) and immunohistochemical technique, respectively.@*RESULT@#Protein kinase CK2alpha mRNA and protein expressions were significantly decreased in tumors transfected with siRNA expression plasmid specific to protein kinase CK2alpha (P<0.05). The tumor grew slowly after transfected with siRNA expression plasmid specific to protein kinase CK2alpha (P<0.01).@*CONCLUSION@#The siRNA expression plasmid specific to protein kinase CK2alpha may suppress the growth and the protein kinase CK2alpha expression of subcutaneous tumors. RNA interfering technology may be a new strategy for the treatment laryngeal cancer.
Subject(s)
Animals , Female , Humans , Mice , Casein Kinase II , Genetics , Cell Line, Tumor , Gene Expression Regulation, Neoplastic , Mice, Inbred BALB C , Mice, Nude , Plasmids , RNA, Small Interfering , Genetics , Transfection , Xenograft Model Antitumor AssaysABSTRACT
In order to study the effect of 5, 6-Dichloro-1-beta-D-ribofuranosyl-benzimidazole (DRB) on the biological characteristics of human laryngeal carcinoma Hep-2 cell line in vitro, Hep-2 cells cultured in vitro were treated with different concentrations of DRB. Changes in cell proliferation, apoptotic rate and invasiveness were detected by MTT assay, flow cytometry (FCM) and matrigel in vitro invasion assay, respectively. It was found that DRB inhibited the proliferation of Hep-2 cells in a dose-and time-dependent manner. After being treated with 0, 10, 20, 40, 80 microm mol/L DRB for 24 h, the apoptotic rate in Hep-2 cells was (0.68+/-0.19)%, (1.95+/-0.12)%, (8.51+/-0.26)%, (11.26+/-0.17)% and (14.99+/-0.32)%, respectively. The matrigel in vitro invasion assay revealed that DRB began to inhibit the invasion of Hep-2 cells at the concentration of 5 microm mol/L, and with the increase of DRB concentration, the inhibitory effect was enhanced. It was suggested that DRB could influence the essential biological characteristics of Hep-2 cells, inhibit Hep-2 cells proliferation, reduce invasive ability and induce apoptosis of Hep-2 cells.
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In order to study the effect of 5, 6-Dichloro-1-β-D-ribofuranosyl-benzimidazole (DRB) on the biological characteristics of human laryngeal carcinoma Hep-2 cell line in vitro, Hep-2 cells cultured in vitro were treated with different concentrations of DRB. Changes in cell proliferation, apoptotic rate and invasiveness were detected by MTT assay, flow cytometry (FCM) and matrigel in vitro invasion assay, respectively. It was found that DRB inhibited the proliferation of Hep-2 cells in a dose- and time-dependent manner. After being treated with 0, 10, 20, 40, 80 μmmol/L DRB for 24 h, the apoptotic rate in Hep-2 cells was (0.68±0.19) %, (1.95±0.12)%, (8.51 ±0.26)%, (11.26±0.17) % and (14.99±0.32)%, respectively. The matrigel in vitro invasion assay revealed that DRB began to inhibit the invasion of Hep-2 cells at the concentration of 5 μmmol/L, and with the increase of DRB concentration, the inhibitory effect was enhanced. It was suggested that DRB could influence the essential biological characteristics of Hep-2 cells, inhibit Hep-2 cells proliferation, reduce invasive ability and induce apoptosis of Hep-2 cells.
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OBJECTIVE@#To purify P0 protein from guinea pig's inner ear by preparative SDS-PAGE and study the possible role it may play in the etiology of autoimmune inner ear disease.@*METHOD@#A mixture of membraneous proteins of inner ear was separated by preparative SDS-PAGE. The corresponding band at 30kd was cut and electrically eluted. The protein collected was identified by analytical SDS-PAGE and Western blot assay. A group of 20 guinea pigs were immunized with P0 protein emulsified in complete Freund's adjuvant, another 10 guinea pigs were immunized with complete Freund 's adjuvant only as control. The guinea pigs' hearing thresholds, serum IgG level and morphological changes in the inner ear were investigated. The distribution of P0 protein in the cochlear was detected by immunohistochemical technique.@*RESULT@#The purity of the protein was demonstrated by a single band at the 30 kD site in SDS-PAGE, which was identified as P0 protein by western blot analysis assay. About 17.5% P0-immunized guinea pigs showed increased hearing thresholds, elevated IgG level (F =6.48, P <0. 01), as well as a decreased number of spiral ganglion cells and inflammatory cell infiltration in the cochlear nerve region. The P0 protein is distributed in the cochlear nerve and spiral ganglion only.@*CONCLUSION@#P0 protein from guinea pig's inner ear can be successfully purified by preparative SDS-PAGE and an animal model of experimental autoimmune inner ear disease induced by P0 protein is successfully established.
Subject(s)
Animals , Adenoviridae , Genetics , Cochlea , Metabolism , Disease Models, Animal , Electrophoresis, Polyacrylamide Gel , Gene Expression , Gene Transfer Techniques , Genes, Homeobox , Genes, Reporter , Genetic Vectors , Guinea Pigs , Myelin P0 ProteinABSTRACT
In order to study the expressions of vascular endothelial growth factor (VEGF) and cyclooxygenase-2 (COX-2) in human laryngeal squamous cell carcinoma (LSCC) and its significance, the expression of VEGF mRNA and COX-2 mRNA in 62 cases of LSCC and 54 adjacent noncancerous laryngeal tissues and 9 normal human laryngeal mucous tissues was detected by using techniques of semi-quantitative RT-PCR. It was found that the expression level of VEGF and COX-2 mRNA was significantly increased in LSCC as compared with that in the normal human laryngeal mucous tissues (both P < 0.01), and the expression level of VEGF and COX-2 mRNA were significantly increased in stage Ill + IV tissues of LSCC as compared with the stage I + II tissues of LSCC (P < 0.01). There was a high positive correlation between VEGF and COX-2 expression in LSCC (r = 0.756, P < 0.01). These data raise the possibility that VEGF and COX-2 may play key roles in the growth, invasion and metastasis of LSCC.
Subject(s)
Carcinoma, Squamous Cell/metabolism , Cyclooxygenase 2/biosynthesis , Cyclooxygenase 2/genetics , Laryngeal Neoplasms/metabolism , RNA, Messenger/biosynthesis , RNA, Messenger/genetics , Biomarkers, Tumor , Vascular Endothelial Growth Factor A/biosynthesis , Vascular Endothelial Growth Factor A/geneticsABSTRACT
In order to study the expressions of vascular endothelial growth factor (VEGF) and cyclooxygenase-2 (COX-2) in human laryngeal squamous cell carcinoma (LSCC) and its significance,the expression of VEGF mRNA and COX-2 mRNA in 62 cases of LSCC and 54 adjacent noncancerous laryngeal tissues and 9 normal human laryngeal mucous tissues was detected by using techniques of semi-quantitative RT-PCR. It was found that the expression level of VEGF and COX-2 mRNA was significantly increased in LSCC as compared with that in the normal human laryngeal mucous tissues (both P＜0.01), and the expression level of VEGF and COX-2 mRNA were significantly increased in stage Ⅲ + Ⅳ tissues of LSCC as compared with the stage Ⅰ + Ⅱ tissues of LSCC (P ＜0. 01). There was a high positive correlation between VEGF and COX-2 expression in LSCC (r=0. 756,P＜0.01). These data raise the possibility that VEGF and COX-2 may play key roles in the growth, invasion and metastasis of LSCC.
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OBJECTIVE To construct siRNA eukaryotic expression vector targeting protein kinase CK2?and to investigate its inhibitory effect on invasion of the HEp-2 cell line in human laryngeal carcinoma. METHODS siRNA expression vector psiRNA-hH1neo-CK2 targeting protein kinase CK2?was constructed by gene recombination,and then was transfected into the HEp-2 cells by lipofectamine methods. Protein kinase CK2?mRNA and protein of the transfected cells were detected by reverse transcription polymerase chain reaction(RT-PCR) and Western blot,respectively. The invasion of the transfected cells was measured by Boyden chamber.SP method was used to examine the expressions of MMP2 and TIMP2 protein of the transfected HEp-2 cells. RESULTS Protein kinase CK2?siRNA expression vector was successfully constructed by gene recombination. Compared with non-specific interfering groups and blank groups, protein kinase CK2?mRNA and protein were significantly decreased respectively in the psiRNA-hHIneo-CK2 groups(P