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Objective:To evaluate the effect of exosomes derived from bone mesenchymal stem cells (BMSCs-EXO) on the postoperative cognitive function and silent infomation regulator 1 (SIRT1)/ nuclear factor kappa B (NF-κB) signaling pathway in aged mice.Methods:BMSCs-EXO were isolated by differential centrifugation method and then identified. Twenty healthy male C57BL/6 aged mice, aged 18 months, weighing 35-40 g, were divided into 4 groups ( n=5 each) using a random number table method: sham operation group (Sham group), operation group (O group), BMSCs-EXO group and EX527 (SIRT1 inhibitor)group. The abdomen regions were shaved for sterilization without exploratory laparotomy in Sham group. Exploratory laparotomy was performed in O group. BMSCs-EXO 50 μg was injected through the tail vein at 1 h before surgery in BMSCs-EXO group. EX527 5 mg/kg was intraperitoneally injected daily at 1-3 days before surgery, and BMSCs-EXO 50 μg was injected through the tail vein at 1 h before surgery in EX527 group. Morris water maze test was used to evaluate the learning and memory ability for 5 consecutive days staring from the 1st day after surgery. Mice were sacrificed at 1 h after the end of Morris water maze test on day 5 after surgery, and the hippocampal tissues were collected for observation of the pathological changes of hippocampal CA1 region and for determination of the expression of tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6) and IL-1β mRNA (quantitative real-time polymerase chain reaction) and SIRT1 and NF-κB p65 (by Western blot). Results:Compared with Sham group, the escape latency was significantly prolonged, the times of original platform crossing were decreased, the swimming time spent in the original platform quadrant was shortened, the expression of tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6) and IL-1β mRNA was up-regulated, the SIRT1 expression was down-regulated, the NF-κB p65 expression was up-regulated ( P<0.05), and the pathological changes of hippocampal tissues in CA1 region were found in O group. Compared with O group, the escape latency was significantly shortened, the times of original platform crossing were increased, the swimming time spent in the original platform quadrant was prolonged, the expression of TNF-α, IL-6 and IL-1β mRNA was down-regulated, the expression of SIRT1 was up-regulated, the expression of NF-κB p65 was down-regulated ( P<0.05), and the pathological changes of hippocampal tissues in CA1 region were significantly attenuated in BMSCs-EXO group ( P<0.05). Compared with BMSCs-EXO group, the escape latency was significantly prolonged, the times of original platform crossing were decreased, the swimming time spent in the original platform quadrant was shortened, the expression of TNF-α, IL-6 and IL-1β mRNA was up-regulated, the SIRT1 expression was down-regulated, the NF-κB p65 expression was up-regulated ( P<0.05), and the pathological changes of hippocampal tissues in CA1 region were accentuated in EX527 group. Conclusions:BMSCs-EXO can improve the postoperative cognitive function in aged mice, and the mechanism may be associated with the activation of SIRT1/NF-κB signaling pathway.
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Objective To study computer toxicity prediction technology and predict the acute toxicity of Chinese materia medica; To provide a new way and method for safety evaluation of traditional Chinese medicine. Methods First, Mold2 software (version 2.0.0) was used to calculate molecular descriptors of 7409 chemical components. After preliminary screening of molecular descriptors, quantitative structure-activity relationship (QSAR) models were built up with Random Forest (RF) for screening the optimum prediction model. From the 83 kinds of toxic Chinese materia medica in Chinese Pharmacopoeia (2010 edition), acute toxicity of 60 kinds of Chinese materia medica reported from monomer structure (1692 chemical components) were under prediction.Results Totally 7409 pieces of data were obtained. When the descriptors were 52, RF modeling accuracy and Kappa were the highest, 0.712 and 0.436 respectively. Compound clusters were divided into 3 types according to optimum molecule descriptors (52). The accuracy and Kappa of the optimum model for the first type of compounds were 0.666 and 0.476 respectively; the accuracy and Kappa of the optimum model for the second type of compounds were 0.804 and 0.381 respectively; the accuracy and Kappa of the optimum model for the third type of compounds were 0.709 and 0.373 respectively. It was predicted that 60 kinds of Chinese materia medica containing 0 violent toxic compound, 2 high toxic compounds, 172 medium toxic compounds and 1518 low toxic compound.Conclusion QSAR model for prediction study on acute toxicity of chemical components of Chinese mareria medica can provide references combination medication and experimental studies.
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In order to provide a new way and method for safety evaluation of Chinese materia medica (CMM) and also to provide a reference for conventional animal experiments, computer toxicity prediction technique and method were established to predict the cardiotoxicity of CMM. Mold2 software (version 2.0.0) was used to calculate molecular descriptors of 1034 chemical components. Then, the random forest (RF) method and the support vector machine (SVM) method were used to screen the descriptors. After that, boosting trees method, SVM, regularized discriminant analysis method and RF method were used to build up prediction model, respectively. Finally, the cardiotoxicity of chemical components was predicted by the quantitative structure-activity relationship (QSAR) model with the best accuracy and Kappa value. The results showed that by comparing the accuracy and Kappa value of prediction model, it was found that the RF model was the optimal algorithm model with 86.3%accuracy and the Kappa value of 0. 725. Through the prediction research on chemical components of Chinese herbs with toxicity recorded in the Pharmacopoeia of People’s Republic of China (version2010),suchasEvodia rutaecarpa,North bean root,Murraya incense,some meaningful results had been received. It was concluded that QSAR model on prediction research of chemical components of Chinese herbs provided important references for further experimental studies and clinical researches.
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Objective To detect the gene expression difference between gastric cancer tissue and lymph node metastasis,screen different express genes,and study its mechanism of metastasis and the relationship with biological behavior.Methods Using U133plus 2.0 gene chip technology,we detected the gene expression difference between gastric cancer tissue and epithelial cells of lymph node metastasis in five patients,and screened out the differentially expressed gene Twist-l.In vitro,the cell proliferation and apoptosis level were measured by using gene over-expression and gene knockout.Metastasis ability of carcinoma cells was detected by cell scratch assay.The expression changes of epithelial-mesenchymal transition (EMT)-associated protein (E-cadherin,Vimentin) in Twist-1 overexpression and gene knockout cells were determined by Western blot.Finally,we detected the expression of Twist-1 in gastric cancer tissue,and its correlation with TNM stage was analyzed.Results The expression of Twist-1 was higher in epithelial cells than in gastric cancer tissue (12.12±3.21 vs.2.07±0.71,P＜0.01).There was no correlation of the expression of Twist-1 with cell proliferation(absorbance of cell proliferation:negtive control 0.84±0.16,null vector control 0.74 ±0.06 and Twist-1 expression cell 0.71 ± 0.07) and apoptosis [cell apoptosis rate:negtive control (2.05±0.08)％,null vector control (4.31±0.07)％ and Twist-1 expression (3.95±0.09)％],but cell migration ability enhanced.In Twist-1 over-expression group,the level of E-cadherin was decreased,while vimentin increased.Conclusions Twist-1 gene changes might be correlated with the metastasis of gastric cancer by the way of EMT.
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Objective To explore the expression of tyrosine phosphatase containing c-Src homology SH2 (SHP-2) in condyloma acuminatum. Methods Phosphorylated tyrosine and SHP-2 were detected in skin lesions of 30 patients with condyloma acuminatum, tumor tissues of 13 cases of cervical cancer and normal foreskin tissues of 11 normal controls by SP immunohistochemical stain. Results In the foreskin tissue, the phosphorylated tyrosine and SHP-2 were mainly expressed on the nuclei of kerotinocytes adjacent to the basal layer, and the number of cells with positive expression was not so many. In condyloma acuminatum, the positive cells arranged in patches, phosphorylated tyrosine and SHP-2 distributed in the cytoplasm. In cervical cancer, the expression of phosphorylated tyrosine and SHP-2 were mainly in the cytoplasm, diffusely distributed, and not in the nuclei. The positive expression rates of phosphorylated tyrosine and SHP-2 in condyloma acuminatum and cervical cancer were 96.67% (29/30) and 100% (13/13), respectively. However the cytoplasm of normal foreskin showed negative expression(0/11). Conclusions There is an aberrant activation of phosphorylated tyrosine and SHP-2 in condyloma acuminatum and cervical cancer. SHP-2 may play an important role in the carcinogenesis.
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Objective To evaluate reliability of the cloned homogeneous immunoassay technique for detection of P53 protein content in serum and stool from patients with gastric carcinoma. Methods To prepare EA, ED and ED P53 fusion protein of beta galactosidase by PGEX 4T 2 expression vector, a novel cloned homegeneous immunoassay was establishment for detection of P53 protein in serum and stool samples from patients with gastric carcinoma and non gastric carcinoma groups as well as healthy control. Results The concentration of P53 protein in 31 gastric carcinoma subjects were remarkably increased as compared with 56 non gastric carcinoma ( P
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<p><b>OBJECTIVE</b>To explore the characteristics of Helicobacter pylori (H. pylori) antigen in serum and to evaluate its clinical diagnostic value.</p><p><b>METHODS</b>Enzyme-linked immunosorbant assay (ELISA) was developed to detect the soluble H. pylori antigen (S-Hp) and circulatory specific H. pylori antigen immunocomplexes (Hp-IC) in serum.</p><p><b>RESULTS</b>The positive rate of S-Hp was 90.91% from 66 patients with H. pylori infection, which was much greater than 0% found in 28 controls (P < 0.001). Moreover, its concentration closely reflected the number of H. pylori in the gastric mucosa layer. We also found that Hp-IC existed bound with IgG and/or IgA in patients with positive S-Hp. However, there is no evidence to show the concentration of S-Hp reduced significantly in followed-up subjects after effective therapy.</p><p><b>CONCLUSIONS</b>These methods as newly and noninvasive complementary tools can be used for clinical diagnosis of H. pylori infection. In addition, S-Hp and Hp-IC may be of importance in H. pylori pathogenesis.</p>
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Adult , Female , Humans , Male , Antibody Specificity , Antigen-Antibody Complex , Blood , Antigens, Bacterial , Blood , Helicobacter Infections , Allergy and Immunology , Helicobacter pylori , Allergy and Immunology , Immunoglobulin A , Blood , Immunoglobulin G , BloodABSTRACT
Objective To study whether omeprazole induces gastric epithelial cells apoptosis and analyse the possible mechanism. Methods The cell cycle and apoptosis ratio of SGC 7901 cell line were determined by flow cytometry in the presence of omeprazole, meanwhile, TUNEL positive cells from gastric mucosa were counted after incubated with omeprazole for 72 hours. Expression of p53、 GADD 45 、 p15,16,18,19 and bcl 2 were stained by immunohistochemical assay and ATPase activity was tested. Results significant alteration with decreasing percentage of G 1 phase and increasing percentage of G 2 phase of SGC 7901 cell line could be detected after incubating 24 hours with omeprazole at double clinical doses . An apoptosis peak appeared in flow cytometry after incubating 72 hours with omeprazole. However, no genetic products such as p53, GADD 45 , p15,16,18,19 and bcl 2 were seen. Concentrations of ATPase were markedly increased from 0.326 0.387 before incubated with omeprazole to 0.054 0.103 after incubated with omeprazole. TUNEL positive reaction cell counts in fresh mucosa was also increased after incubated with omeprazole for 72 hours( P
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Objective:Measured the soluble forms of intercellular adhesive molecule 1 (sICAM 1), soluble vascular cell adhesion molecule ( sVCAM 1) and interleukin 1(IL 1),tumor necrosis factor(TNF), interferon ?(IFN ?),study the role of sICAM 1 and sVCAM 1 in rheumatoid arthritis(RA).Methods:A sandwich ELISA technique was used to measure the serums levels of sICAM 1, sVCAM 1 ,IL 1,TNF,IFN ? in 30 patients with active RA and 30 normal control subjects.Results:The levels of sICAM 1 and sVCAM 1 in patients with RA were significantly higher than that in control subjects(P
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0.05),However,the amount of metronidazole efflux was significantly decreased after adding verapamil(P
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Objective To explore the anti-proliferation and apoptosis-induced effects of matrine on HT29 cell and their possible mechanism.Methods HT29 Cell proliferative activity was measured by MTT assay;Cell cycle and apoptosis were analyzed by Flow Cytometry;Alteration of cellular skeleton was observed by transmission electron microscopy(TEM);Global gene expression profiles were scanned by gene chip.Results After exposure to matrine at concentrations from 0.062 5 to 0.5 mg/mL for 48 h,cellular proliferation was inhibited with increasing the concentration,but this inhibitory effect attenuated at 1 mg/mL and the apoptosis was up-regulated significantly.G2/M and G1/S phases of cell cycle were,to some extent,arrested.Under TEM,the morphological changes could be found.Gene chip showed that matrine could alter a large number of genes,which related to the cell proliferation,cell cycle,and apoptosis.Conclusion The anti-proliferation and apoptosis-induced effects of matrine on HT29 cells are concentration-dependent through changing of many genes involved in proliferation,cell cycle,and apoptosis.