Frequency characteristics of horizontal semicircular canals damage and the ultrastructure analysis of crista ampullaris in patients with Meniere's disease / 中华耳鼻咽喉头颈外科杂志

Objective: To investigate the frequency characteristics and the pathological characteristics of the horizontal crista ampullaris in patients with Meniere's disease,and to analyse its structural basis. Methods: Between March, 2019 and November, 2019, seventy-two patients diagnosed as Meniere's disease (27 males and 45 females, aged from 13 to 74 years, with a course of disease ranging from 4 months to 32 years)in Shandong Provincial ENT Hospital were included.Caloric test, sinusoidal harmonic acceleration test (SHA), video-head impulse test (v-HIT), Gadolinium-enhanced inner-ear 3D-FLAIR MRI and pure tone audiometry were conducted in the patients. The function of the horizontal semicircular canal in these patients were analysed as well as its relationship with the degree of endolymphatic hydrops,clinical stage and duration. Light microscopy and transmission electron microscopy were used to observe the ultrastructure of horizontal semicircular canal crista ampullaris from six patients with refractory Meniere's disease who underwent labyrinthectomy. The number of type Ⅰ and type Ⅱ vestibular hair cells, the common pathophysiological changes of horizontal semicircular canal crista ampullaris were investigated in these patients. Statistical analysis was performed using SPSS 19.0. Results: With the increase of detection frequency, the abnormal rate decreased gradually. The abnormal rate of caloric test was 69.4% (50/72), SHA 51.4% (37/72), V-HIT 36.1% (26/72), comparation of the positive rate among the three tests showed statistically significant differences(P<0.05).Neither caloric test nor SHA had correlation with the degree of hydrops(P>0.05), but v-HIT(r=0.434,P<0.01).There was correlation with clinical stage to SHA and v-HIT(r=0.338,0.462,P<0.01), except caloric test(P>0.05).No significant relation was found with caloric test, SHA, v-HIT and course of disease(P>0.05).Morphological observation found abnormal monolayer epithelialization of the horizontal semicircular canal crista ampullaris significantly decreased number of type Ⅱ hair cells compared with type Ⅰhair cells. Hair cells showed perinuclear vacuolization, cytoplasmic vacuoles, mitochondrial electron density increasement and loss of stereocilia. Conclusions: The horizontal semicircular canal damage in the patients with Meniere's disease has a frequency-dependent characteristic, mainly occurres in low frequency area. With progress of the disease, the high frequency area of ampulla will be impaired gradually, and it is related to the degree of endolymphatic hydrops and hearing level. Hair cell injury would be observed,the frequency characteristics may be more associated with the disorder of type Ⅱ hair cells.

Impact of clopidogrel on gene profile of human umbilical vein endothelial cell line and bioinformatics analysis / 中国实验血液学杂志

This study was purposed to investigate the effect of clopidogrel on gene expression profile of cultured human umbilical vein endothelial cell (HUVEC) line (EA.hy926), and explore its potential molecule mechanism. A Affymetrix U133 plus 2.0 oligonucleotide microarray was applied to detect the alteration of gene expression profile induced by clopidogrel in HUVEC. Real time RT-PCR was used to verify the result of selected differentially expressing genes. The results showed that total 508 genes (including 139 up-regulated and 369 down-regulated genes) were obtained with differential expression more than 1.5-fold after clopidogrel (10 µmol/L) acted on HUVEC for 48 h. Clopidogrel affected the expression levels of genes involved protein binding, transcription factor activity, zinc ion binding, regulation of DNA-dependent transcription, transcription, RNA splicing and so on. It is concluded that the clopidogrel modulate function of endothelial cells by regulating sets of genes through different pathway.

Impact of clopidogrel carboxylic metabolite SR26334 on gene expression profile of human umbilical vein endothelial cell line / 中国实验血液学杂志

This study was purposed to characterize the effect of carboxylic acid metabolite (SR26334) of clopidogrel bisulfate deprived of antiplatelet efficacy on the spectrum of gene expression in the cultured human umbilical vein endothelial cell (HUVEC) line (EA.hy926), and to explore the potential molecule mechanism of SR26334 impact on HUVEC. By using a Affymetrix HU133 plus 2.0 oligonucleotide microarray, the alteration of gene expression spectrum induced by SR26334 in HUVEC was detected, the real-time PCR was used to confirm the results of selected differentially expressing genes. The results indicated that total 235 including 176 up-regulated and 59 down-regulated genes were obtained with change more than 1.5-fold after SR26334 (10 µmol/L) acted on HUVEC for 48 h. SR26334 affected the expression levels of genes involved regulation of transcription, transcription, positive regulation of transcription from RNA polymerase II promoter, cell cycle, cell division, protein amino acid dephosphorylation in HUVEC. It is concluded that carboxylic acid metabolite SR26334 of clopidogrel bisulfate modulates function of endothelial cells through different pathway at gene level.

Effect of irbesartan on the proliferation, apoptosis and VEGF mRNA expression of human umbilical vein cell line ea. hy926 in vitro / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To evaluate the effect of irbesartan on the proliferation, apoptosis, and VEGF mRNA expression of human umbilical vein cell line EA.hy926 in vitro.</p><p><b>METHODS</b>The human umbilical vein cell line EA.hyY926 were treated with various concentrations of irbesartan for 24 h. The cell proliferation after the treatment was detected by CCK8 assay, flow cytometry and FITC Annexin V/PI kit were used to detect changes in the cell apoptosis. RT-PCR was used to evaluate the expression of VEGF mRNA.</p><p><b>RESULTS</b>There were no changes in cell shape with various concentration of irbesartan. CCK-8 assay showed a greater rate of the cell proliferation in irbesartan group than that in control group with a dose-independent manner after 24 h treatment. After incubation with irbesartan, cell proliferation rate was significant (P < 0.05). FCM analysis showed no significantly changes in the cell apoptosis. Irbesartan increased the proliferation of EA.hy926 cells. At concentration of 1 x 10(-4), 1 x 10(-5), 1 x 10(-6) mol/L, VEGF mRNA expression enhanced either (P < 0.05).</p><p><b>CONCLUSION</b>Irbesartan could promote the proliferation and up-regulated VEGFmRNA expression in EA.hy926 cell line. This result suggested that in addition to antihypertensive effect, angiotensin receptor antagonist might be a novel therapeutic approach to chronic ischemic heart disease as heart failure.</p>

Angiogenesis related gene expression profiles of EA.hy926 cells induced by irbesartan: a possible novel therapeutic approach / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Angiogenesis occurs commonly in various physiological and pathological processes. Improving blood supply through promoting angiogenesis is a novel approach for treating ischemic diseases. Angiotensin II type 1 receptor blockers (ARBs) dominate the management of hypertension, but evidence of their role in angiogenesis is contradictory. Here we explored the angiogenic effects of ARBs through characterizing gene expression of the human umbilical vein endothelial cell line EA.hy926 exposed to irbesartan.</p><p><b>METHODS</b>The human umbilical vein endothelial cell line EA.hy926 was grown for 72 hours after treatment with different concentrations of irbesartan. The cell proliferative capacity was assessed by CCK8 assay at 24, 48 and 72 hours. Gene expression levels in EA.hy926 cells responding to irbesartan were measured under optimal proliferation conditions by microarray analysis using Affymetrix U133 plus 2.0. The differential expression of genes involved in angiogenesis was identified through cluster analysis of the resulting microarray data. Quantitative RT-PCR and Western blotting analyses were used to validate differential gene expression related to the angiogenesis process.</p><p><b>RESULTS</b>In the 10(-4), 10(-5), 10(-6) mol/L treatment groups, cell proliferation studies revealed significantly increased proliferation in EA.hy926 cells after 24 hours of irbesartan treatment. However, after 48 and 72 hours of treatment with different concentrations of irbesartan, there was no significant difference in cell proliferation observed in any treatment group. We selected the group stimulated with irbersartan at a concentration of 10(-6) mol/L for microarray experiments. Statistical analysis of the microarray data resulted in the identification of 56 gene transcripts whose expression patterns were significantly correlated, negatively or positively, with irbesartan treatment. Cluster analysis showed that these genes were involved in angiogenesis, extracellular stimulus, binding reactions and skeletal system morphogenesis. Of these 56 genes we identified seven genes (VEGF, KDR, PTGS2, PLXND1, ROBO4, LMO2, and COL5A1) involved in the angiogenesis process. qRT-PCR analysis of these genes confirmed the microarray results. Protein expression of three VEGF pathway genes (VEGF, KDR, and PTGS2) was further confirmed by Western blotting.</p><p><b>CONCLUSIONS</b>Our study showed that irbesartan may induce angiogenic effects in vascular endothelial cells. It suggested that the mechanism of angiogenic effects of ARBs might be attributed to the signaling cascade from angiotensin receptors in the VEGF pathway. It also provided evidence indicating that ARBs could be used as a novel therapeutic approach to treat chronic ischemic heart disease as well as anti-hypertensive agents.</p>

Comparison on the effects of clopidogrel, statins combination in treating coronary artery disease among the elderly patients: a retrospective cohort study / 中华流行病学杂志

Objective To compare the effects of clopidogrel with or without combined with CYP3A4-metabolized statin in treating coronary artery disease (CAD) among the elderly patients.Methods The study cohort was defined as all patients were over 60 years of age and hospitalized for CAD who were prescribed clopidogrel between January 2000 and February 201 1.A total of 1021 patients were enrolled,with 178 of them prescribed clopidogrel and 843 patients were administrated clopidogrel combined with statins (CYP3A4-metabolized statins 636 and non CYP3A4-metabolized statins 207).The primary endpoint was all cause of death and the second endpoint were non-fatal myocardial infarction (MI),but hospitalized for unstable angina,stroke,transient ischemic attack,or repeated revascularization (PCI or coronary artery bypass graft).Results Among the clopidogrel group and the clopidogrel plus statins group,the incidence density of death was 6.86/1000 and 3.18/ 1000 respectively,with crude RR as 2.15(95％CI:1.39-3.33) and statistically significant (x2=3.53,P＜0.01).The incidence density of composite thromboembolic events did not show statistical significance (P＞0.05).The two groups were 1∶1 matched,after propensity score matching,clopidogrel coadministrated with statins group showed significant decrease in all cause of death,with RR as 0.42 (95％ CI:0.19-0.93),x2=7.23,P＜0.01.No significant difference was observed in deaths or composite thromboembolic events between statins via different cytochrome P450 pathways.Conclusion Clopidogrel with statin could reduce the mortality of elderly CAD patients compared with clopidogrcl without statin.The result did not show statistical significance between CYP3A4-metabolized statins or non CYP3A4-metabolized statins regarding the mortality or composite endpoint events.

Irbesartan regulates inflammatory gene expressions related to atherosclerosis in EA.hy926 cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To characterize if irbesartan regulates vascular inflammatory gene expression profiles related to atherosclerosis in EA.hy926 cells.</p><p><b>METHODS</b>Human umbilical vein endothelial cell line EA.hy926 cultured in vitro was incubated with irbesartan (1×10(-6) mol/L) for 24 h. The total RNA was extracted from the cells for gene expression profiling. The DAVID Gene Functional Classification Tool was used to analyze the disease- and function-related genes in the cells. Real-time quantitative polymerase chain reaction (RT-PCR) was used to verify the genes showing differential expression after irbesartan treatment. The protein levels of angiotensin II type 1 receptor (AT1R) and type 2 receptor (AT2R) were tested by Western blotting.</p><p><b>RESULTS</b>Compared with the control cells, 56 genes were found to show marked changes following irbesartan treatment, including 39 up-regulated and 17 down-regulated genes. Disease analysis suggested that these genes were related to such diseases as coronary atherosclerosis, myocardial infarction, and colorectal cancer. Eight genes, namely MMP2, PTGS2, PECAM1, SELP, SELL, CYP1A1, MMRN1, and HSPA1A, were involved in atherosclerosis and myocardial infarction. Verification by RT-PCR produced a result consistent with the gene array result. AT1R was down-regulated while AT2R up-regulated in irbesartan-treated cells.</p><p><b>CONCLUSION</b>Irbesartan regulates the inflammatory gene expressions related to atherosclerosis in EA.hy926 cells. These inflammatory factors may promote destabilization of atherosclerotic plaque possibly in relation to AT2R overexpression.</p>

Establishment of animal model of breast cancer dormancy / 中华病理学杂志

<p><b>OBJECTIVE</b>Tumor dormancy has been defined clinically as a condition in which tumor cells are present but do not grow for a long period of time. Breast cancer is noted for its long periods of tumor dormancy and metastases can occur many years after treatment.</p><p><b>METHOD</b>Simulating the characteristics of breast cancer patients after treatment, we established the animal model of breast cancer dormancy by inoculating 500 Ca761-03 cells into the limb muscle of 615 mice and then selecting animals with tumor dormancy 2 months post inoculation (corresponding to 5 years for humans).</p><p><b>RESULTS</b>Two months after inoculation of Ca761-03 cells into the muscle of 615 mice, tumor occurred in 30% of the mice. The remaining 70% of mice did not show tumor growth. After repeated traumatic stimulation, 90% of the mice developed tumors after 5 months, therefore representing tumor dormancy.</p><p><b>CONCLUSIONS</b>These results demonstrate that breast cancer cells can remain in a dormant state for long periods of time in vivo. Trauma can stimulate the dormant tumor cells to proliferate again, and causes tumor relapse. This murine model system promises a sound animal model for the study of solid tumor dormancy.</p>

Homocysteine and Mild Vascular Cognitive Impairment / 国际脑血管病杂志

Hyperhomocysteinemia is an important independent risk factor for cardio-cerebrovas- cular diseases.Studies in recent years have suggested that hyperhomocysteinemia is also an independent risk factor for vascular dementia and Alzheimer's disease.Mild vascular cognitive impairment is the prophase of vascular dementia.Mainly through the damage of vascular wall structure and its function,homocysteine may has correlation with mild vascular cognitive impairment.

Establishment and characterization of a rabbit tumor cell line VX2 / 中华病理学杂志

<p><b>OBJECTIVE</b>To establish a rabbit tumor cell line and to characterize its biological parameters.</p><p><b>METHODS</b>VX2 tumor tissue was used for the primary culture in vitro. After 40 passages, the cell morphology, CK expression (immunohistochemical staining), cell cycle, karyotype and tumorigenecity in rabbits and nude mice were investigated.</p><p><b>RESULTS</b>The newly established cell line VX2 was maintained in continuous culture for over 70 passages in 10 months. Morphologically, VX2 cells were polygonal to short spindled. Tonal fibril and tight junction were found under the electron microscope. CK was positive. The cell cycle analysis showed 69.3% in G1 phase, 5.6% in G2 phase and 25.1% in S phase. The population doubling time was 34.5 hours. The chromosomal analysis showed a hypotriploidy with a median chromosome number of 58 approximately 62. The tumorigenecity in rabbits and nude mice were both 100%.</p><p><b>CONCLUSION</b>The established VX2 cell line derived from rabbit squamous carcinoma could serve as a model system for experimental oncology in the rabbit.</p>

Clinical study of effect of tianzhi granule on senile vascular demetia / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the clinical effect of Tianzhi Granule (TZK) on senile vascular dementia (VaD), which is classified as sthenia of liver-yang.</p><p><b>METHOD</b>Two hundred VaD patients were treated with TZK (0.5 g/bag), which was taken one bag each, three times a day. The treatment course was one month and they were treated for rwo courses.</p><p><b>RESULT</b>TZK could remarkably increase gnosia and activity, with no striking difference from that of positive control group (P > 0.05). Simultaneously, TZK could significantly improve the clinical syndrome of traditional Chinese medicine and viability. It could also drastically reduce the whole blood and plasma viscosity and improve erythrodegeneration and abnormality of aggregation index in the abnormal blood viscosity patients.</p><p><b>CONCLUSION</b>TMC has certain effects on senile VaD.</p>