AIM:ToobservetheeffectofcepharanthineonhumanlungadenocarcinomaLTEP-a-2cellgrowth, and to explore the changes of related microRNA ( miRNA) expression in the cells .METHODS:LTEP-a-2 cells were trea-ted with cepharanthine at concentrations of 0μmol/L, 10μmol/L, 20μmol/L and 40μmol/L.The growth inhibition rate was detected by MTT assay , and the cell morphological changes were observed under light microscope .The cell apoptosis was analyzed by flow cytometry .The expression of let-7c, miR-34a and miR-34b was measured by real-time PCR.RE-SULTS:Cepharanthine inhibited the cell activity of LTEP-a-2 cells in a dose-dependent manner .With the increase in cepharanthine concentration , the pyknosis of the cells was visible under the inverted microscope .Flow cytometry analysis found that different concentrations of cepharanthine induced the increase in the apoptotic rates of LTEP -a-2 cells.The re-sults of real-time PCR showed that the cepharanthine also increased the expression of let -7c, miR-34a and miR-34b.CON-CLUSION:Cepharanthine inhibits the growth of LTEP-a-2 cells, and induces apoptosis .Cepharanthine increases the ex-pression of let-7c, miR-34a and miR-34b, indicating that these miRNAs in LTEP-a-2 cells has the function as tumor sup-pressor genes .

Objective: To observe ascitic interleukin-7 expression level in cirrhotic patients complicated with spontaneous bacterial peritonitis, and to detect the effect of recombinant human IL-7 on CD4⁺ and CD8⁺T lymphocyte function. Methods: A total of 84 patients with liver cirrhosis who were hospitalized from August 2017 to April 2018 were selected. Among them, 51 cases were complicated with cirrhosis and untainted ascites, and 33 cases were cirrhosis complicated with spontaneous bacterial peritonitis. Peripheral blood and ascites were collected routinely. The levels of IL-7 in peripheral blood and ascites were measured by enzyme-linked immunosorbent assay. CD4⁺T cells and CD8⁺T cells were purified from ascites, and were stimulated with recombinant IL-7. Cellular proliferation, key transcription factors for mRNA, and cytokines production by CD4⁺T cells in response to IL-7 stimulation was measured. mRNA expression corresponding to perforin, granzyme B, and granulysin as well as cytokines production by CD8⁺T cells was also measured in response to IL-7 stimulation. Cytolytic and non-cytolytic activity of CD8⁺T cells in response to IL-7 stimulation was also investigated in both direct and indirect contact co-culture system. Measurement data of the normal distribution were compared between the two groups by Student’s t-test and the data before and after stimulation were compared by paired t-test. Measurements that did not conform to normal distribution were compared between the two groups using Mann-Whitney U test, and data before and after stimulation were compared using Wilcoxon paired test. Results: There was no significant statistical difference in serum IL-7 levels between the two groups [(5 001 ± 1 458) pg/ml vs. (4 768 ± 1 128) pg/ml, P = 0.41]. The level of ascitic IL-7 in cirrhotic patients complicated with SBP was significantly lower than cirrhosis patients with untainted ascites [(966.4 + 155.8) pg/ml vs. (792.1 + 126.4) pg/ml, P < 0.01]. Recombinant IL-7 stimulation promoted the proliferation of CD4⁺ and CD8⁺T cells from ascites in patients with liver cirrhosis complicated by SBP. T-bet mRNA relative expression and IFN-γ secretion in CD4⁺T cells was also elevated in response to IL-7 stimulation in vitro. Moreover, IL-7 stimulation also increased the mRNA expressions of perforin, granzyme B, and granulysin as well as productions of IFN-γ and TNF-α by CD8⁺T cells. Recombinant IL-7 stimulation elevated cytolytic and non-cytolytic activity of CD8⁺T cells from ascites in patients with liver cirrohosis complicated by SBP, which manifested as increased target cell death and IFN-γ production in both direct and indirect contact co-culture system. Conclusion Ascitic IL-7 promotes T lymphocyte function in patients with liver cirrhosis complicated with SBP.

Objective To investigate the residents' acceptance and the influencing factors of the hierarchical medical system in Xuzhou, and to suggest on effective system implementation. Methods Xuzhou residents free of cognitive impairment and over 18 years old were sampled for questionnaire survey in July-August 2016, to study their acceptance of their basics and acceptance of the system. 1 550 questionnaires were distributed, and 1 473 valid ones were recovered. The count data were expressed as constituent ratio, and χ2test was used for single-factor analysis, with binary logistic regression analysis for multi-factor analysis. Results 71. 0% of the residents embraced this system. Their acceptance varies significantly with their age, place of residence, education, annual average monthly income, self-rated health status, physical examination experience, conditions of chronic diseases, medical visit experience at primary healthcare institutions, and their awareness of the system (P<0.05). Conclusions The acceptance of the system by Xuzhou residents needs to be elevated, by means of greater promotional efforts, capacity building for primary institutions, so as to fully leverage the system to serve the residents.

Objective:To investigate the effect of astragaloside A (AS-A) on the photoreceptor degeneration induced by sodium iodate (NaIO 3) and its related mechanism. Methods:Sixty healthy male C57BL/6J mice, aged 6-8 weeks, were randomly divided into normal control (NC) group, NaIO 3 group, and ASA group, with twenty mice in each group. 30 min before modeling, AS-A group mice were intraperitoneally injected with 100 μl AS-A at a dose of 100 mg/kg body weight. 30 min later, mice in NaIO 3 group and AS-A group were intraperitoneally injected with 100 μl NaIO 3 at a dose of 30 mg/kg body weight. Subsequently, AS-A group mice were administered AS-A twice daily at 12 h intervals until the end of the experiment. On day 1 post-modeling, zonula occludens-1 (ZO-1) immunohistochemistry was performed to observe the structure of retinal pigment epithelium (RPE) cells; real-time quantitative polymerase chain reaction (qPCR) was conducted to detect the mRNA expression of various retinal chemokine ligand-2 ( Ccl2), interleukin-1 beta ( Il-1β), mixed lineage kinase domain-like protein ( Mlkl), receptor-interacting protein kinase 3 ( Ripk3), and tumor necrosis factor ( Tnf). On day 3 post-modeling, immunohistochemistry was performed to observe the expression of ionized calcium binding adaptor molecule 1 (Iba1) and glial fibrillary acid protein (GFAP) in the retina; TdT-mediated dUTP nick-end labeling (TUNEL) assay was used to detect photoreceptor cell death in each group. On day 4 post-modeling, fundus morphology of mice in each group was observed by fundus color photography and optical coherence tomography (OCT). Hematoxylin-eosin staining (HE) was used to observe the morphological structure of the retina in each group. Inter-group comparisons between two groups were conducted using independent samples t-test, while comparisons among three groups were performed using one-way ANOVA. Results:Fundus color photography and OCT examination showed that a large number of scattered yellow-white subretinal nodular structures in the fundus of NaIO 3 group mice, and a large number of strong reflection areas in the RPE layer. The number of strong reflection areas in the RPE layer was reduced in the AS-A group. Immunohistochemical analysis of ZO-1 showed that ZO-1 was largely lost on the RPE cell membrane in that NaIO 3 group; whereas in the AS-A group, ZO-1 was evenly distributed on the RPE cell membrane. HE staining results showed circular black deposits were visible in the RPE layer of the NaIO 3 group, and the inner and outer segments of photoreceptors were severely damaged, with a significant decrease in the number of outer nuclear layer (ONL) cell nuclei; whereas in the AS-A group, the RPE layer pigments were orderly, the inner and outer segments of photoreceptors were intact, and the number of ONL cell nuclei significantly increased. The results of TUNEL staining show that numerous TUNEL-positive cell nuclei were observed in the ONL of the retina in the NaIO 3 group, while the number of TUNEL-positive cell nuclei in the ONL of the retina was significantly reduced in the AS-A group, with statistically significant differences ( t=2.66, P<0.05). The analysis of qPCR data showed that compared with the AS-A group, the relative expression levels of Mlkl, Ripk3, Ccl2, Il-1β and Tnf mRNA in the retina were significantly increased in the NaIO 3 group, with statistically significant differences ( F=39.18, 10.66, 53.51, 41.40, 24.13; P<0.001). Immunohistochemical staining results showed that compared with NC group and AS-A group, the positive expression of GFAP in retina of NaIO 3 group was significantly increased, and the difference was statistically significant ( F=9.62, P<0.05). Conclusion:AS-A antagonizes NaIO 3-induced photoreceptor degeneration in part by inhibiting photoreceptor cell death and neuroinflammation. Meanwhile, AS-A treatment protects against NaIO 3-triggered perturbation of retinal homeostasis.

Signal transducer and activator of transcription 3 (STAT3) and Chemokine CX3C ligand 1 (Fractalkine/CX3CL1) play important roles in vascular inflammation and injury. To study if STAT3 promotes vascular endothelial cell proliferation and migration through fractalkine, we overexpressed or knocked down STAT3 in vascular endothelial cells, and used quantitative real-time PCR and Western blotting to determine the effect of STAT3 on fractalkine expression. The wild type and STAT3 binding site mutant fractalkine promoter luciferase reporter plasmids were constructed, and luciferase activity assays were used to explore the effect of STAT3 on the transcriptional activity of the fractalkine promoter. MTT assays were used to detect the effect of overexpression or knockdown of STAT3 or fractalkine on the proliferation rate of vascular endothelial cells. Scratch assays were used to detect the effect of overexpression or knockdown of STAT3 or fractalkine on vascular endothelial cell migration. There results showed that overexpression of STAT3 could promote fractalkine expression, and knockdown of STAT3 could down-regulate fractalkine expression. STAT3 could directly bind to the promoter of fractalkine to promote its transcriptional activity via binding the GAS site of the fractalkine promoter. Knockdown of STAT3 could inhibit the migration of vascular endothelial cell, and overexpression of fractalkine antagonized this inhibition. Our data concluded that STAT3 promotes the proliferation and migration of vascular endothelial cell by binding the GAS site of the fractalkine promoter to promote fractalkine transcriptional activity and expression.
Cell Proliferation ; Chemokine CX3CL1 ; Endothelial Cells ; Promoter Regions, Genetic ; STAT3 Transcription Factor

Objective : To explore the relationship between geriatric nutritional risk index( GNRI) and perioperative nutritional status，postoperative recovery and complications in elderly patients with gastric cancer． Methods : In this retrospective study，212 elderly patients ( aged ≥60 years ) with gastric cancer who underwent gastrectomy were recruited．GNRI was used to retrospectively assess the patients' preoperative nutritional status ，and analyze the relationship between GNRI and perioperative nutritional status，postoperative recovery and complications．The ROC curve was applied to explore the value of GNRI in predicting postoperative complications. Results : The inci- dence of preoperative nutritional risk in elderly patients undergoing gastric cancer surgery was 45. 07%．Compared with the patients whose GNRI＞98 points，the patients whose GNRI≤98 points had different degrees of decrease in serum total protein，albumin，prealbumin，hemoglobin and lymphocyte counts before surgery，day 1 and day 5－8 after surgery (P ＜0. 05) ．The patients whose GNRI ＜92 points had longer postoperative hospital stay than those with GNRI＞98 points (P＜0. 05) ．With the decrease of GNRI scores，the incidence of complications showed an upward trend(P＜0. 001) ．The multivariate analysis of the relationship between GNRI and postoperative complica- tions showed that TNM staging of III －IV and GNRI ＜92 points were independent risk factors for complications. GNRI had a good predictive value for the occurrence of complications (AUC = 0. 639，95% CI : 0. 570－0. 703，P = 0. 001，Cut-off value : 92. 21) ． Conclusion GNRI can be used for preoperative nutritional assessment for eld- erly gastric cancer patients．Patients with GNRI＜92. 21 points should be actively given nutritional therapy to im- prove perioperative nutritional status，speed up postoperative recovery，and reduce the occurrence of complications.