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Objective Salidroside is a major active component of integripetal rhodiola herbal medicine, which has a significant activity against hypoxia and ischemia.This study was to investigate the effects of side chain carbon losssalidroside analogues (N04) on the expressions of HIF-1α-related factors in the hypoxia-injured EAhy926 human vascular endothelial cells.Methods EAhy926 human umbilical vein endothelial cells in the logarithmic growth phase were randomly divided into a normal control, a hypoxia model control, a salidroside, a high-dose N04, a medium-dose N04, and a low-dose N04 group.The hypoxia model was established by depriving the culture medium of sugar and serum and culturing the EAhy926 cells in an environment of 95%N2+5%CO2 for 2 hours, followed by intervention with salidroside at 1×10-6 mol/L and N04 at 1×10-6, 1×10-7, and 1×10-8 mol/L, respectively.Then, the activity of the cells was detected by MTT assay, their LDH activity examined by spectrophotometry, the mRNA expressions of HIF-1α and VEGF measured by RT-PCR, the protein expressions of HIF-1α, VEGF and pVHL determined by Western blot, and the activity of eNOS measured by ELISA.Results Compared with the normal control group, the hypoxia model cells showed significantly reduced activity (0.51±0.05 vs 0.27±0.02, P<0.01), an elevated LDH level ([6.65±1.43] vs [78.82±2.33] U/L, P<0.01), and decreased eNOS activity ([1.56±0.23] vs [1.16±0.20] U/100 mL, P<0.01).In comparison with the hypoxia model group, the cells treated with high-, medium-, and low-dose N04 exhibited remarkably increased activity (0.27±0.02 vs 0.0.42±0.05, 0.40±0.03 and 0.37±0.04, P<0.01), a reduced LDH level ([78.82±2.33] vs [53.05±3.90], [58.42±4.45] and [62.73±3.63] U/L, P<0.01), and increased eNOS activity ([1.16±0.20] vs [3.01±0.47], [2.60±0.26] and [2.32±0.29] U/100 mL, P<0.01).The activity of eNOS was also increased in the salidroside group ([2.32±0.29] U/100 mL, P<0.01).The cell activity in the high-and medium-dose N04 groups was markedly higher than that in the salidroside group (P<0.05), and so was the eNOS activity in the high-dose N04 group and the LDH level in the medium-and low-dose N04 groups (P<0.05).In comparison with the normal control group, the expressions of HIF-1α mRNA, HIF-1α protein and VEGF protein were significantly up-regulated in the hypoxia model group (P<0.01) while that of the pVHL protein markedly down-regulated (P<0.01).Compared with the hypoxia model group, the expressions of HIF-1α mRNA, HIF-1α protein and VEGF protein were remarkably reduced (P<0.05), while that of the pVHL protein markedly elevated (P<0.05).Both the expressions of VEGF mRNA and HIF-1α protein were significantly lower in the medium-and low-dose N04 groups than in the salidroside group (P<0.05).Conclusion N04 can protect vascular endothelial cells against hypoxia-induced injury by regulating the expression of HIF-1α-related factors and eNOS.
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Objective:To study the gene regulatory network during the osteogenic differentiation process of dental follicle stem cells (DFSCs)with bioinformatic analysis.Methods:The differentially expressed genes during the osteogenic diffrentiation process of DF-SCs were selected from the GEO Datasets.Then the relationship among the genes were analysed using DAVID and GeneMANIA data-base.Results:Numerous differentially expressed genes during the osteogenic differentiation process of DFSCs were obtained,289 genes with significance of different expression(P <0.05)were enriched into “cell differentiation”,“cell proliferation”,“skeletal sys-tem development”and “calcium signaling pathway”subgroups.ALPL,CTSK,TUFT1 ,TGFBR2,FHL2,ROR2,STC1 ,POSTN, ZBTB1 6,FRZB,PRELP and IGFBP5 were enriched into the “skeletal system development”subgroup.The 1 2 genes exhibited interac-tions,including co-expression,co-localization,genetic interaction and signal pathways.Conclusion:There is a complex molecular regulatory network among the differentially expressed genes during the osteogenic differentiation process of DFSCs.It is necessary to an-alyse the osteogenic differentiation process of DFSCs as a whole from WNT,TGFbeta,MAPK siganal pathyways,Homeobox transcrip-tion factors and osteogenic differentiation marker genes.
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Severe hand-food-mouth disease can be accompanied by neurogenic pulmonary edema,whicn can rapidly lead to death.The pathogenesis of neurogenic pulmonary edema may be related to the comprehensive influences on neural factors,humoral factors and multiple bioactive substances after the central nervous system is damaged.It is of great impotance to investingate the pathogenesis of neurogenic pulmonary edema,which facilitates timely treatment and prevention of the complications.
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The medical image registration between preoperative three-dimensional (3D) scan data and intraoperative two-dimensional (2D) image is a key technology in the surgical navigation. Most previous methods need to generate 2D digitally reconstructed radiographs (DRR) images from the 3D scan volume data, then use conventional image similarity function for comparison. This procedure includes a large amount of calculation and is difficult to archive real-time processing. In this paper, with using geometric feature and image density mixed characteristics, we proposed a new similarity measure function for fast 2D-3D registration of preoperative CT and intraoperative X-ray images. This algorithm is easy to implement, and the calculation process is very short, while the resulting registration accuracy can meet the clinical use. In addition, the entire calculation process is very suitable for highly parallel numerical calculation by using the algorithm based on CUDA hardware acceleration to satisfy the requirement of real-time application in surgery.
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Humans , Algorithms , Imaging, Three-Dimensional , Tomography, X-Ray Computed , X-RaysABSTRACT
Objective To compare the efficacy and safety of a new low-dose oral contraceptive pill (YAZ) containing drospirenone 3 mg and ethinylestradiol 20 μg with placebo in reducing symptoms of premenstrual dysphoric disorder (PMDD). Methods This multicenter, double-blind, randomized clinical trial consisted of 2 run-in and 3 treatment cycles (84 days) with daily symptom charting; 187 women with symptoms of PMDD were randomized to either placebo group (n=94) or YAZ group (n=93), and assessed with daily record of severity of problems scale (DRSP) and clinical global impressions scale (CGI) before, during and after the treatments. Hormones were administered for 24 days, followed by 4 days of inactive pills. Results Compared with baseline level of DRSP, both groups got improvement after treatment; the YAZ group (median-28.7, range:-82.5 to 2.3) had greater improvement than that in the placebo group (median-23.7, range:-86.0 to 11.8), while there was not significant difference (P>0.05). The main adverse effects of YAZ included intermenstrual bleeding [13% (12/93) versus 3% (3/94)], menorrhagia [9% (8/93) versus 1%(1/94)], nausea [5%(5/93) versus 4%(4/94)] and skin rash [4%(4/93) versus 2%(2/94)]. Conclusions YAZ could improve symptoms of PMDD better than placebo, while without statistic significance in this study. The most common adverse effects are intermenstrual bleeding, menorrhagia, nausea and rash.
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Objective To study in vitro the inhibitory effects and mechanisms of N-butanol extract of Potentilla anserine L.(NP)against hypoxia-induced nitric oxide(NO)in hippocampus neuron of rats. Methods The models of hippocampus neurons hypoxia injury of Sprague-Dawley(SD)neonatal rats were cultured in vitro. The cultured hippocampus neurons were divided randomly into blank control group, hypoxia injury model group, nimodipine group(2 μmol/L)and NP high(250.0 mg/L),middle(62.5 mg/L),low(15.6 mg/L)dose groups. The activities of hippocampus neurons were examined by methyl thiazolyl tetrazolium(MTT)assay,and meanwhile their contents of nitrogen monoxidum(NO)were detected. Half quantity reverse transcription-polymerase chain reaction(RT-PCR)and Western blotting were used to detect neuronal nitric oxide synthetase(nNOS)mRNA and protein expression levels respectively in each group,immunocytochemistry stain was used to detect protein positive rate. Results Compared with blank control group,the activity of neuron〔absorbance(A)value〕was significantly decreased(0.0826±0.0095 vs. 0.3315±0.0105),content of NO(μmol/g:0.0509±0.0027 vs. 0.0291±0.0032), the expression levels of nNOS mRNA (0.1463±0.0081 vs. 0.0801±0.0058), the positive rate of nNOS〔(74.4238±3.9423)%vs.(28.3714±4.1361)%〕,the expression levels of nNOS protein(A value:1.9130±0.0471 vs. 0.5068±0.0368)were all significantly increased in the hypoxia injury model group(all P0.05). Conclusions NP can ameliorate the injury of rat hippocampus neurons induced by hypoxia in vitro. The possible mechanisms might be related to the effective inhibition of the synthesis of nNOS and NO excessive generation.
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Objective To investigate the protective effect of n-butanol extract from the roots of Potentilla anserina (NP) on hypoxic hippocampal neurons in neonatal rats.Methods Primary cultured hippocampal neurons were pretreated with different concentration of NP (0.25,0.0625,and 0.0156 mg/mL) before incubation in a low oxygen (0.1％) environment for 4 h.Cell viability was evaluated by Trypan blue staining assay.Lactate dehydrogenase (LDH) released by neurons into the medium was measured.The activity of superoxide dismutase (SOD) in cell cytosol was determined using nitroblue tetrazolium.Morphological changes and mitochondrial function were observed by transmission electron microscopy.Results Hypoxic injury could decrease the cells viability of neuron,enhance LDH release (P ＜ 0.05),decrease SOD activity,and increase mitochondrial injury.Pretreatment with NP significantly increased cell viability,decreased LDH release (P ＜ 0.05),promoted SOD activity (P ＜ 0.05),and remarkably improved cellular ultra-microstructure compared with the model group.Conclusion NP could protect the primary hippocampal neurons from hypoxic injury by attenuating mitochondrial cell death.
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ObjectiveTo investigate the effect of n-butanol extract from Potentilla anserina (NP) intervention on hypoxia-induced Ca2+ overload and SERCA2 expression of rat cardiomyocytes.MethodsPrimary cultured myocardial cell from SD neonatal rat (1-3 d) was used in the establishment of hypoxia model.After hypoxia for 3 h,the Ca2+ concentration of myocardial cells was measured with fura-2/AM fluorescent probe,and the biochemical indicator intracellular Ca2+-ATPase was examined and the mRNA and its protective protein levels of the sarcoplasmic reticulum (SR) Ca2+-ATPases (SERCA2) were assayed with RT-PCR,Western-blotting,and immune-cytochemical staining in each group.ResultsThe results showed that NP decreased Ca2+ concentration,increased the activity of Ca2+-ATPase,and improved the mRNA and protein expression of SERCA2 in hypoxia-injured myocardial cells as compared with the model group.ConclusionThese results indicate that NP could attenuate the Ca2+ overload.The mechanism might be explained as that NP could elevate the SERCA2 level,increase the activity of myocardium in rats,and further enhance the capacity of SR Ca2+ re-uptake.
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<p><b>OBJECTIVE</b>To observe the protective effect of alcohol extract of Potentilla anserina against myocardial apoptosis induced by acute myocardial ischemia/reperfusion by arteria coronaria ligation and the effect on the expressions of Caspase-3 and Caspase-9 in myocardial apoptosis signal pathway.</p><p><b>METHOD</b>Male SD rats were randomly divided into the sham-operated group, the model group, the diltiazem group (30 mg x kg(-1)) and P. anserine alcohol extract intervention groups (0.9, 1.8, 3.6 g x kg(-1)). Rat acute myocardial ischemia/reperfusion model was established by ligating left anterior descending. Apoptosis of myocardial cells were detected by TUNEL (Terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling assay). The expressions of Caspase-3 and Caspase-9 mRNA were assayed by reverse transcription-polymerase chain reaction (RT-PCR). Semi-quantitative analysis was made for the expressions of Caspase-3 and Caspase-9 by immunohistochemistry.</p><p><b>RESULT</b>According to TUNEL results, after I/R injury-induced myocardial apoptosis, the apoptotic index (AI) of model group was (31.5 +/- 3.6)%. All P. anserine alcohol extract intervention groups showed obvious inhibition of ischemia/reperfusion-induced myocardial apoptosis. In the model group, myocardial apoptosis caused increased expression of Caspase-3, Caspase-9 mRNA and proteins. After the administration of P. anserine alcohol extract, 1.8, 3.6 g x kg(-1) dose groups showed notable decrease in Caspase-9 mRNA (P < 0.05), while the 0.9 g x kg(-1) dose group showed no significant difference with the model group. Alcohol extract of P. anserina in all dosages showed inhibitory effect on the expression of Caspase-3 mRNA in myocardial cells compared with model group (P < 0.05). Immunohistochemistry showed that administration of all dosages of alcohol extract of P. anserina could significantly reduce Caspase-3 and Caspase-9 protein expressions after I/R injury (P < 0.05).</p><p><b>CONCLUSION</b>The administration with alcohol extract of P. anserina can protect the myocardial tissue from apoptosis after acute myocardial ischemia and reperfusion injury in rats and inhibit the expressions of Caspase-9 and Caspase-3 mRNA and proteins.</p>
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Animals , Male , Rats , Apoptosis , Caspase 3 , Metabolism , Caspase 9 , Metabolism , Ethanol , Chemistry , Myocardial Reperfusion Injury , Drug Therapy , Plant Extracts , Chemistry , Therapeutic Uses , Potentilla , Rats, Sprague-DawleyABSTRACT
To investigate the protective effects of scutellarin benzyl ester on neonatal rats' cardiomyocytes injured by ischemia and its anti-apoptosis mechanism.
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BACKGROUND: As the first long-acting atypical antipsychotics, the therapeutic effect and safety of risperidone microsphere have been proved. However, it may lead to serious pain due to the deep intramuscular injection.OBJECTIVE: To evaluate the pain levels by 12-week injection of rispeddone microsphere and to explore the relationship among dose and times of injection of risperidone microsphere and pain levels.METHODS: A total of 57 patients diagnosed as schizoprenia by DSM-Ⅳ, aged 18-65 years, were selected and injected risperidone microsphere once every 2 weeks with doses of 25, 37.5 and 50 mg. The pain levels were evaluated using 100 mm visual analogue scale during injection and at the injected sites. The effects of injected dose, injected frequency and injected sites on the pain were analyzed by the nurse questionnaire.RESULTS AND CONCLUSION: The pain levels among the different doses groups had no notable differences (F=1.35,P> 0.05), which demonstrated that the pain had no relationship with injected dose. However, the pain level of injected sites had correlation to injected doses. The pain level of the 50 mg group was greater than that of the 37.5 and 25 mg groups. Accordingly,patients who treated by high dose of risperidone microsphere should be intervened by nurses.
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Objective To study the mechanism of chemotherapy resistance caused by interleukin-6 (IL-6) in ovarian cancer cells and its related signal pathways. Methods Ovarian cancer cell lines A2780(IL-6 receptor positive, while non-IL-6-expressing and cisplatin/paclitaxel-responsive) and SKOV3 cell lines( overexpressing of IL-6 receptor and IL-6 and cisplatin/paclitaxel-resistant) were suitable models for this study. The effect of exogenous (a short period of treatment with recombination IL-6) and endogenous IL-6(by transfecting with plasmid encoding for sense IL-6 ) in A2780 cells or deleting of endogenous IL-6expression in SKOV3 cells (by transfecting with plasmid encoding for antisense IL-6) on the sensitivity to cisplatin and paclitaxel was investigated. Meanwhile, the mechanism of chemotherapy resistance caused by IL-6 in ovarian cancer cells and its related signal pathways were also analyzed. Results We found that both exogenous and endogenous IL-6 induce cisplatin and paclitaxel resistance in non-IL-6-expressing A2780 cells (the resistance multiple to cisplatin and paclitaxel was: exogenous, 6. 25 and 7.31; endogenous, 7. 13 -8. 34 and 7. 61 - 10. 70), while deleting of endogenous IL-6 expression in IL-6-overexpressing SKOV3 cells promotes its sensitivity to anticancer drugs ( the resistance multiple to cisplatin and paclitaxel was 0. 15 and 0. 10, 0. 10 and 0. 08). IL-6 significantly up-regulated the expression levels of mRNA and protein of drug resistance-associated genes, MDR1 and GST-π, and apoptosis-inhibiting genes, bcl-2, bcl-xL and XIAP in a dose-dependent manner in A2780 cells. In accordance with this finding, the mRNA and protein levels of MDR1 and GST-π enhanced in sense IL-6-transfected A2780 cells, and reduced in antisense IL-6-transfected SKOV3 cells compared with the corresponding parental and control vector-transfected cells, which had no difference. It was found that PD98059 [ mitogen-activated protein kinase-extracellular signalregulated kinase (MEK) inhibitor ] and wortmannin [ phosphatidylinositol 3-kinase (PI3K) inhibitor ]significantly antagonized IL-6-induced phosphorylation of extracellular signal-regulated kinase ( ERK ) and protein kinase B (Akt), respectively, and both of them blocked IL-6-induced cisplatin and paclitaxel resistance and the inhibitory effects of PD98059 and wortmannin were dependent on its concentration.Conclusions These data suggest that IL-6-induced chemoresistance may be associated with increase of both drug resistance-associated genes ( MDR1 and GST-π) and apoptosis-inhibiting genes ( bcl-2, bcl-xL and XIAP), and activation of MEK/ERK and PL3K/Akt. Therefore, modulation of IL-6 expression or its related signaling pathway may be a promising strategy of treatment for drug-resistant ovarian cancer.
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To investigate the protective effects of the n-butanol extract of Potentilla anserina L. (NP) on pituitrin-induced acute myocardial ischemic injury in mice.
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In the reformation,we have applied the technique of multimedia to the experiments,which has made the nonobjective theories simplified and the key and difficult points in teaching aesy to understand and increased the content of teaching in the limited teaching hours.It has proved that we achieved a good effect in teaching.
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AIM: To study the expression of c-fos mRNA in brain following moderate lateral fluid percussion brain injury in rats, and to observe the temporal patterns of its expression following percussion. METHODS: Male Sprague-Dawley rats were divided into normal control, sham operation control and injury group. The rats of injury group subjected to moderate lateral fluid percussion injury (0.2 mPa). The injury groups were then subdivided into 5 min, 15 min, 30 min, 1h, 2h groups according to the time elapsed after injury. The expression of c-fos mRNA was studied with reverse transcription polymerase chain reaction (RT-PCR) semi-quantitatively.RESULTS: At 5 min after percussion, the induction of c-fos mRNA was increased, and remained elevated up to 2 h after brain injury.CONCLUSION: The induction and expression of the c-fos mRNA in cortex and brain stem after fluid percussion brain injury were increased rapidly.
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AIM: To study the expression of c-jun in brain stem following moderate lateral fluid percussion brain injury in rats, and to observe the temporal patterns of its expressions following percussion.METHODS: Male Sprague-Dawley rats were divided into normal control, sham operation control and injury groups. The rats of injury group subjected to moderate lateral fluid percussion injury (0.2 mPa), and then were subdivided into 5 min, 15 min, 30 min, 1 h, 2 h, 4 h, 8 h and 12 h groups according to the time elapsed after injury. The expression of c-jun was studied by immunohistochemistry and in situ hybridization. RESULTS: After percussion for 15 min, Jun positive neurons increased in brain stem progressively, and peaked at 12h. At 5min after percussion, the induction of c-jun mRNA was increased, and remained elevated up to 1h-2h after brain injury. CONCLUSION: The induction and expression of the c-jun in brain stem after fluid percussion brain injury were increased rapidly and lasted for a long time.
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0.05),while the activities of SOD of bone tissue were lower(P
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Proteolysis and amino acid metabolism in autolyzing rabbit left ventricular myocardium were studied. Tissue slices were incubated in a moist chamber at 37℃ from 15 min to 480 min. Their free acidic, neutral and basic amino acids and ammonia were determined by ion exchange chromatography. It was found thatmost of the free tissue amino acids in the course of autolys is increased significantlg, Increasing of alanine, lysine and histidine was noted at 15min of autolysis while the tissue glutamic acid content decreased,and the tissue ammonia content increased. The results suggest that the protcelysis was accelerated in early autolyzing heart muscles. It is helpful for the timing of death.
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The early histological myocardial ischemic changes in both experimental rabbitsand forensic caseworks of sudden death were demonstrated by the hematoxylin basicfuchsin-picric acid staining method.After the ligation of the coronary artery,therewere discrete patchy fuchsin-stained fibers which in contrast with the normal fibersof light-brown color.The number of fuchsin-stained fibers increased with the pro-longation of the intervals after ischemia.The results were not affected by the post-mortem autolysis.We consider that this method can be used as a routine test ofearly myocardial ischemia in both clinico-pathology and forensic pathology.