Objective To investigate the effects of periplocin from Cortex Periplocae (CPP) on apoptosis of human lung cancer A549 cells and expression of survivin, and demonstrate its anti-tumor effect and the possible mechanism. Methods Inhibitory effect of CPP at different concentrations (1. 25, 2. 50, 5. 00, 10. 00, 20. 00 ng·mL-1 ) and for different time length (24, 48, 72 h) on A549 cell proliferation was tested by MTT method. Apoptosis rate of A549 cells treated with CPP at different concentrations (2. 50, 5. 00, 10. 00 ng·mL-1 ) were measured using flow cytometry (FCM) for 6, 12, 24, 48, 72 h, respectively. The morphological and ultrastructural changes of the apoptosis cells were observed by acridine orange/ ethidium bromide (AO/ EB) staining and transmission electron microscopy (TEM). The effects of CPP on mRNA and protein expression of apoptosis associated gene survivin were assessed by RT-PCR and Western blotting. Results CPP could significantly inhibit the growth of A549, and the inhibition rate reached (93. 46±2. 35)% . The results of FCM showed that the apoptosis rate of A549 cells treated with CPP was increased significantly as compared to the control group ( P<0. 05). Meanwhile, typical apoptotic peaks were detected. The characteristic morphological changes of apoptosis were observed in A549 exposed to CPP, including cell shrinkage, the nuclei became yellow-red by AO/ EB staining, and typical ultrastructural changes, including nuclear chromatin condensation along the nuclear membrane, vacuolar degeneration of cytoplasm observed by TEM. The result of RT-PCR indicated that survivin mRNA expression decreased obviously in A549 cells exposed to CPP. The protein expression of survivin in A549 cells treated with 10. 0 ng·mL-1 CPP(0. 251±0. 012)was weaker than that in control group(0. 928±0. 016). Conclusion CPP can induce apoptosis in human lung cancer cell lines A549, and the probable mechanism is related to the down-regulation of survivin mRNA and protein.

Objective To investigate the inhibitory effects of periplocin from cortex periplocae (CPP) on human lung cancer cell line QG56 and to discuss its mechanism. Methods QG56 cells were cultured in vitro. The final concentrations of CPP in control group were 1.25, 2.50, 5.00, 10.00 and 20.00μg/L. QG56 cells were treated with ascending concentration of CPP for 24 h, 48 h and 72 h. The cell proliferation was measured using MTT method. The morphological changes of QG56 cells were observed under inverted microscope. Flow cytometry (FCM) was used to detect the effects of CPP on cell cycle and cell apoptosis. The expression of apoptosis associated gene bax mRNA in QG56 cells was detected by RT-PCR. The expres-sion of bax protein before and after treatment of CPP was examined by SP immunocytochemistry. Results The inhibitory ef-fect of CPP on the proliferation of QG56 cells was increased with the increasing concentrations of CPP and the prolonged du-ration of treatment. The morphological changes were displayed in QG56 exposed to CPP. The results of FCM showed that CPP caused cell cycle arrest at G0/G1 phase. The apoptotic rate of QG56 cells was significantly increased after CPP treatment for 48 h (P<0.05). The expression of bax mRNA was increased in QG56 exposed to CPP. The result of immunocytochemis-try indicated that CPP up-regulated the expression of bax protein. Conclusion CPP showed significant inhibitory effect on human lung cancer cell lines QG56 through inducing cell cycle arrest and apoptosis.

[ ABSTRACT] AIM:To observe the inhibitory effect of madecassoside on the LPS-stimulated microglia and to inves-tigate its possible mechanism.METHODS:Microglia cells of neonatal Sprague-Dawley ( SD) rats were cultured, isolated and purified.Microglia cells were activated with lipopolysaccharide ( LPS) .The inhibitory effect of madecassoside on micro-glia was measured by MTT assay.Tumor necrosis factor alpha (TNF-α), interleukin 1β(IL-1β) were detected by ELISA. Cell cycle and apoptotic rate were evaluated by flow cytometry.The expression of TLR4 was detected by Western blotting. The expression of NF-κB was detected by RT-PCR.RESULTS: LPS induced the proliferation of microglia and release in-flammatory cytokines significantly.Compared with LPS group, madecassoside inhibited the proliferation of microglia induced by LPS in a dose dependent manner.The IC50 value of madecassoside was 10.97 nmol/L to microglia after incubation for 48 h.Madecassoside also decreased the levels of TNF-αand IL-6, increased the ratios of microglia at the G2 phase and the ap-optotic rate, decreased the expression of TLR4 and NF-κB significantly (P<0.05).CONCLUSION:Madecassoside has in-hibitory effects on the proliferation of LPS-stimulated microglia, by which the mechanism may be related to inhibition of the expression of TLR4 and NF-κB, change of cell cycle distribution and induction of microglia apoptosis.

Objective To investigate the effect and possible mechanism of high mobility group box (HMGB) 1 in the development and progress of rheumatoid arthritis.Methods PBMC and serum samples were obtained from 74 RA patients (38 in active stage and 36 in stable stage) and 26 healthy controls.The expression of HMGB1 mRNA and protein was detected by RT-PCR and ELISA.Flow cytometry analysis ( FCM ) was used to detect the expression of Toll-like receptor 4 on PBMC.Results ①The expression of HMGBI mRNA and protein in active RA patients was significantly higher than that in healthy controls and inactive RA patients [2.63 vs 0.71，0.93 and (10.2±1.2) vs (7.5±1.8)，(8.3±1.8) ng/ml，respectively](P＜0.01 ).② The relative expression of TLR4 protein on CD14+ monocytes and CD3+ lymphocytes in active RA patients was increased than that in inactive RA and healthy controls (P＜0.05 or P＜0.01 ).It was also higher in inactive RA than in healthy controls (P＜0.05 or P＜0.01 ).③ Level of HMGB1 protein in serum of RA patients was positively correlated with ESR，CRP，RF，the numbers of tender joints and swollen joints as well as radiographic changes.Conclusion HMGB1 can be synthesized and released by PBMC of active RA patients，and then bind to TLR4 of PBMC to promote inflammatory responses and bone erosion.

Objective To investigate the effect of tail-suspension,noise stress and combination of them on immune function in rats.Methods The rats were divided into four different groups,A: control;B: noise stress(2 h);C: tail-suspension(30?,21 d);D: tail-suspension added noise stress.The body weight,thymus weight and spleen weight,circulating leukocytes and its distribution,T lymphocyte subpopulations(with flow cytometry) and NO level in serum(with Griess method) of rats were determined after exposed to 21 d tail-suspension,2 h noise stress and combination of them.Results Compared with group A,the body weight were significantly decreased(P

Objective:To understand the reliability and validity of quality of life instruments for cancer patients-brain neoplasm [QLICP-BN (V1.0)], a self-developed quality of life scale for cancer patients.Methods:The quality of life of 112 patients with brain neoplasms in Yunnan Cancer Hospital from March 2012 to November 2013 was measured. The general data questionnaire and QLICP-BN (V1.0) were used for data collection. The reliability, validity and responsiveness of the scale were tested, and then the metric characteristics of the scale were evaluated.Results:The split-half reliability of the total score of the scale was 0.95, the Cronbach αcoefficient was 0.92, and the test-retest correlation coefficient rwas 0.78. After extracting common factors by the principal component method and rotating with the maximum variance, the specific module obtained three principal components, and the cumulative variance contribution rate was 64.18%. The score of specific module was 75.30±17.44 before treatment and 78.91±12.20 after treatment ( t=-2.481, P=0.015). The total score of scale before treatment was 65.26±12.29, and that after treatment was 69.62±10.41, with a statistically significant difference ( t=-4.492, P<0.001). The total responsiveness of the scale was 0.456, showing moderate responsiveness. Conclusion:QLICP-BN (V1.0) has good reliability, validity and a certain degree of responsiveness. It can be used as a measurement tool for the quality of life of patients with brain neoplasms in China.

OBJECTIVETo explore the molecular events and mechanism in the carcinogenesis of esophageal epithelium in the high incidence area of esophageal carcinoma.

METHODSEpithelial cells collected from the high incidence area of esophageal carcinoma were used to detect DNA content and ploidy by propidium iodide(PI) stain. The expressions of p53, p16 and cyclin D1 were stained by indirect immunofluorescence of fluorescein isothiocyanate(FTTC), which were detected by flow cytometry (FCM).

RESULTSDuring the process of carcinogenesis, DNA content increased significantly. The diploid cells decreased while heteroploid cells increased sharply, with a heteroploidy rate of 84.2%. At the same time, the p53 protein accumulated and p16 was deleted. The positive rates of p53 and oncogene cyclin D1 were both 100%(5/5, 6/6) in the cancer group.

CONCLUSIONIn the early carcinogenesis of esophageal epithelium, DNA content and heteroploidy rates increase with tumor suppressor gene p16 deletion and p53 protein accumulation while oncogene cyclin D1 is overexpressed. Multiple molecular events have already occurred when esophageal carcinoma develops.

Cyclin D1 ; metabolism ; Cyclin-Dependent Kinase Inhibitor p16 ; metabolism ; DNA, Neoplasm ; metabolism ; Esophageal Neoplasms ; genetics ; metabolism ; pathology ; Flow Cytometry ; Gene Expression ; Humans ; Precancerous Conditions ; genetics ; Tumor Suppressor Protein p53 ; metabolism

Objective: To compare the reliability of Internet Addiction Impairment Index (IAII), Revised Chen Internet Addiction Scale(CIAS-R)-Taiwan Revision, CIAS-R-Mainland Revision, Young Diagnostic Questionnaire (YDQ) and the consistency of Internet addiction using the four scales in college students. Methods: A total of 1 004 undergraduates from 3 universities in Hefei were selected to measure the tendency of internet addiction simultaneously using the four scales, and 122 students were re tested two weeks after the initial assessment. Correlation coefficient, coincidence rate and Kappa value were used to analyze the consistency of the four scales. Analysis of variance, t test and Logistic regression were used to determine the consistency of the factors related to internet addiction scale. Results: The reliability of the four Internet addiction scales were greater than 0.7( P <0.01). The correlation coefficient among all scales was greater than 0.5( P <0.01). The agreement between YDQ and CIAS-R-Mainland Revision was 0.87. The Kappa value of YDQ and CIAS-R-Taiwan Revision in the consistency analysis was 0.51( P <0.01), the Kappa value between the other scales was less than 0.5. Results showed that the four scales were consistent in Internet addiction prevalence by gender, grade and major, while CIAS-R-Taiwan Revision and YDQ were not consistent with the other two scales in sleep disorder. Conclusion The four Internet addiction scales all have good reliability, while low agreement in Internet addiction assessment, suggesting further improvement and revision in Internet addiction scales.

Objective: To explore the mediating role played by college students social anxiety and social support in the relationship between internet addiction and depression. Methods: A cluster random sampling method was used to conduct a questionnaire survey among 3 536 college students in three higher vocational colleges in Anhui Province. The content included general demographic characteristics, depression, Internet addiction, social support, and social anxiety. The Process program was used to mediate and analysis of regulation. Results: Among the survey subjects, 1 552(43.90%) had depressive symptoms, including 561(45.65%) boys and 991(42.96%) girls.The total score of Internet addiction was significantly positively correlated with depression score(r=0.30, P<0.01); social anxiety(social fear, social avoidance) was positively correlated with depression(r=0.24, 0.27, P<0.01); social support(subjective support, objective support, support utilization) was significantly negatively correlated with depression(r=0.25, -0.23, -0.17, P<0.01). Conditional process analysis shows that social anxiety had a mediating role between internet addiction and depression(c'=0.06, P<0.01), and that Internet addiction and social anxiety were regulated by social support(β=-0.00，P=0.02). Conclusion By increasing the social support of college students to improve social anxiety, it might help to reduce the depression of college students caused by Internet addiction.

Objective:To test whether the constructed intervention model of advance care planning (ACP) for patients with advanced cancer can be successfully implemented and the preliminary intervention effect, which provides reference for empirical research.Methods:32 cases of advanced cancer patients and 25 cases of their families at the Fifth Affiliated Hospital of Zunyi Medical University, Zhuhai were selected. The patients were subdivided into the experimental group and the control group by random number table method. The control group received routine nursing, while the experimental group adopted the intervention model of "VIP for future care" on the basis of customary nursing. We measured the main outcome indicators: enrollment rate, consent rate, completion rate and loss of follow-up rate and secondary outcome indicators: decision-making certainty, end-of-life care preference and post-intervention satisfaction of patients and their families, within 1 week and 1 month after intervention.Results:The enrollment rate, consent rate and loss of follow-up rate were 74.6% (206/276), 36.9% (76/151)and 15.6% (5/32), respectively. After intervention, the completion rates of the experimental group and the control group were 16/16 and 15/16 within one week, and 14/16 and 13/16 within one month. All of the family members were conducted during the follow-up period. The intervention mode of "VIP for future care" had a statistically significant difference in decision-making certainty between the two groups of patients ( β=0.63, 95% CI 0.08-1.18, P<0.05), no statistically significant difference in end-of-life care preference between the two groups of patients and their families ( P>0.05), and had a statistically significant difference in "whether to recommend this project to others" between the two groups ( χ2 value was 4.167 , P<0.05). Conclusions:On the premise of sufficient preparation, the "VIP for future care" intervention mode can be successfully implemented in advanced cancer patients in mainland China, can improve the decision-making certainty of patients and the satisfaction of patients and their families, and it is recommended. And should be applied to ACP intervention for patients with advanced cancer.