Objective To investigate the effect and mechanism of emodin inducing the apoptosis of human bile duct carcinoma cell QBC939 in vitro. MethodsThe inhibition of cell proliferation was detected by MTT. The morphological changes of apoptotic cells were observed under fluorescence microscopy.The apoptosis rates and intracellular reactive oxygen species levels were detected by flow cytometry.The intracellular relative activities of caspase-9 and caspase-3 were tested through colorimetric method.ResultsEmodin inhibited cell proliferation of human bile duct carcinoma cell QBC939 in time-dose dependent fashion.Apoptotic cells displayed bright,nuclear presented divided leaves,debris and a set of edges under fluorescence microscopy.Treated with 30 μmol/L and 50 μmol/L emodin,the apoptosis rates of 24 hours were respectively 38.9％ ± 9.07％ and 67.09％ ± 4.08％ (P ＜ 0.05).The intracellular reactive oxygen species levels after 30 minutes treatment were 1.65 ± 0.08 and 2.28 ± 0.04folds of the control group (P ＜ 0.05). Emodin could activate caspase-9 and caspase-3,leading to elevations of their activities (P ＜ 0.05).ConclusionEmodin inhibites cell proliferation of human bile duct carcinoma cell QBC939 through inducing apoptosis.The mechanism is associated with the elevated levels of reactive oxygen species and the activation of caspase-9 cells and caspase-3.

In the occurrence and development of tumors, side population cells play an extremely important role. They have the characteristics of cancer stem cells, especially their potential of tumor originating, and stronger drug-resistance. The study of biological characteristics, sorting and training methods of side population cells and the relationship between side population cells and tumor drug-resistance may shed some light on the diagnosis and treatment of cancer.

Objective To observe the effects of the different serum uric acid levels on expression of Alzheimer’s disease biomarkers (APP and BACE1) in rats. Methods Intraperitoneal injection of oxygen of oxazine acid potassium was used to produce HUA models in rats. H&E staining was used to detect the morphological changes of the hippocampus. Western blot was used to detect the protein levels of APP and BACE1 of the hippocampus. Results Compared with nor-mal control group, the serum uric acid and the protein levels of APP and BACE1 in the hippocampus was obviously in-creased at OAPS treatment group (P<0.01). Compared with low dose OAPS treatment group, the serum uric acid level was significantly increased whereas the protein levels of APP and BACE1 in the hippocampus were significantly decreased at the middle and high dose group (P<0.01). Compared with middle dose group, the serum uric acid level was increased at high dose group (P<0.05) and the expression of APP were decreased in the hippocampus rats but the expression of BACE1 remained unchanged (P>0.05). Conclusion The higher level of serum uric acid may be a protective factor of AD. The higher serum uric acid levels, the lower the risk of AD.

Objective To observe the influence of p53-upregulated modulator of apoptosis (PUMA) on the growth of human brain glioma cell lines U251 (p53 mutant) and SHG-44 (p53 wild type),and to explore its possible mechanism.Methods Construct the adenovirus PUMA (Ad-PUMA) and vector of adenovirus (AdDsRed) which were respectively transfected into glioma cell lines U251 and SHG-44.Cells proliferation rates were measured with cell counting kit-8 (CCK-8).The apoptotic ratios were detected by flow cytometry.The expression of PUMA and apoptosis associated proteins (bcl-2,Bax) were determined with Western blot analysis.Caspase-3,Caspase-8,Caspase-9 activity were measured by Caspase activity assay kit.Results Compared with vector group and blank control group,Ad-PUMA transfected group showed strong cell proliferating inhibition effects [the inhibition rates were (50.89±4.73) ％ and (44.45±5.33) ％ respectively,P ＜0.05] and pro-apoptotic effects [apoptotic rates were (44.89±5.08) ％ and (31.67±7.32) ％,P ＜ 0.05] in different p53 glioma cell lines U251 and SHG-44.Western blot analysis showed that PUMA protein expression increased after Ad-PUMA transfection,accompanied by the reduced expression of the anti-apoptotic protein bcl-2 and the increased expression of pro-apoptotic protein Bax.The activity of Caspase testing results showed that the Caspase-3,Caspase-9 activity increased significantly,while the Caspase-8 activity changed little.Conclusion No matter how p53 phenotype,PUMA can inhibit glioma proliferation,promote apoptosis,and its mechanism may be through the mitochondrial apoptotic pathways,upregulation of Bax and inhibition of bcl-2 expression,which activated Caspase-9.Ad-PUMA is expected to become a new target for gene therapy of gliomas.

Objective To discuss the effect of comprehensive mental intervention on mental health and sleep quality among the crew members during the Escort Mission in the Gulf of Aden.Methods The same batch of two escort ships was divided into study group and control group.The study group was intervened by comprehensive psychological intervention methods involving diversified forms of psychological training, counseling, behavioral intervention and cooperation between various departments.A psychological survey was conducted with Symptom Checklist 90( SCL-90) , Self-rating Anxiety Scale (SAS), Self-rating Depression Scale(SDS) and Pittsburgh Sleep Scale(PSQI) before and 3 months after intervention. Results There was no significant difference between the two groups before intervention (P>0.05).After intervention,the score of SCL-90 of study group was significantly better than that of the control group in somatization,obsessive compulsive disorder,interpersonal relationship,depression, anxiety,bigotry and hostility (P<0.05), while the score of SAS and SDS was lower(P <0.05).Sleep quality, the time taken to fall asleep, sleep time, dyssomnia,sleep efficiency,daytime dysfunction factor and the total score of PSQI were significantly more desirable than in control group (P<0.01).Conclu-sion Comprehensive intervention can effectively improve mental health and sleep quality of the crew members in the escort mission in the Gulf of Aden.It is worthy of popularization and application.

Objective:To investigate the effect and mechanism of targeted B7-H3 gene silencing on the tumorigenesis and metastasis of human hematological malignancy xenograft tumor in nude mice.Methods: Real-time fluorogentic quantitative PCR (qPCR) and flow cytometry (FCM) were used to detect the expression of B7-H3 in 13 strains of malignant hematologic cells.Then,U937,Maver and Z138 cells which expressed high level of B7-H3 were screened out.Targeted B7-H3 knockdown in U937,Ma-ver and Z138 was performed by lentivirus transduction and the effect of B7-H3 silencing in stable cell lines was tested by qPCR and FCM.Injecting the nine groups subcutaneously into the nude mice to establish xenograft models after dividing the U937,Maver and Z138 into non-infected control group (CON),B7-H3 knockdown group (KD) and negative non-targeted control infected group (NC),respectively,for detecting the tumorigenicity and metastasis in vivo.Furthermore,the expression of Ki-67 in xenograft tumors was detected by immunohistochemistry (IHC).The expression of metalloproteinase 2 (MMP-2) was detected by western blot.Results: The stable B7-H3 silencing cell lines of U937,Maver and Z138 were successfully established.Compared with the NC group,the KD groups of U937,Maver and Z138 had an obviously slower tumor growth.The average tumor inhibition rates at the end of observation period were 61.83％ (F=43.78,P<0.05),59.12％ (F=36.51,P<0.05) and 67.37％ (F=40.29,P<0.05);there was no significant difference in tumor volume growth between the NC group and the CON group (P>0.05).The liver distant metastasis of all the xenograft tumor models in nude mice was the most common and the rates of distant metastasis in KD groups were significantly lower than that of the corresponding NC groups.The Ki-67 indexes of the KD groups were significantly lower than those of the relative NC groups in three cell lines (U937: 40.3％±5.2％ vs.79.1％±6.3％,q=30.31,P<0.05,Maver: 35.2％±6.4％ vs.69.6％±5.1％,q=24.82,P<0.05;Z138: 38.4％±7.1％ vs.75.7％±4.8％,q=28.07,P<0.05);there was no significant difference in the expression of Ki-67 between the NC group and the CON group (P>0.05).The expressions of MMP-2 were also significantly lower in the KD groups than in the NC groups (U937: q=14.59,P<0.05;Maver: q=9.25,P<0.05;Z138: q=11.04,P<0.05);there was no significant difference in the expression of MMP-2 between the NC group and the CON group (P>0.05).Conclusion: Targeted B7-H3 gene silencing could inhibit the tumorigenesis and metastasis of human hematological malignancy xenograft tumor in nude mice.The mechanism may be related to the down-regulation of Ki-67 and MMP-2.

Objective To investigate radiation-induced alternations of phospholipids in epithelial cells,and to provide experimental evidence for understanding the mechanism of radiation-induced intestinal injury.Methods The intestinal epithelial cells(IEC-6)in rats were divided into three groups:normal control group,8 Gy X-ray irradiation group and 12 Gy X-ray irradiation group.Phospholipids were extracted at 6 h or 24 h after radiation and then measured by high-performance liquid chromatography and mass spectrometry(HPLC-MS).Results At 6 h after radiation,the phospholipids in 8 Gy irradiation group didn't vary significantly,while those in 12 Gy irradiation group changed.The PG,PI and Lyso PC were significantly up-regulated(F=5.37,9.60,9.88,P＜0.05).However,at 24 h after radiation,many PE and PG species in both irradiation groups declined(F=5.15-99.77,P＜0.05)and SM species increased in 12 Gy irradiation group(F=4.35-7.92,P＜0.05).Conclusions The ionizing radiation could disorder phospholipid metabolism in IEC-6 cells with a dose-dependent manner.

AIM: To investigate the effects of stress on the opening of mitochondrial membrane permeability pore (PTP) in rat heart and explore the possible molecular mechanism underlying PTP opening. METHODS: Stress animal model was established. After strained for differnet time, all rats were killed and PTP opening degree were examined by spectrophotometer. Bcl-2, Bax expression levels were determined by Western blot. RESULTS: Stress induced PTP opning, Bcl-2 expression inhibition and Bax level elevation in myocardial mitochondria. CONCLUSION: PTP opening was the important mitochondrial mechanism of stress-induced heart injury. Decrease in Bcl-2 expression and increase of Bax level may be an important molecular basis for PTP opening.

Histology and Embryology is a very important basic curriculum of medicine. It is directly related to the freshmen’s interest in learning medicine continually. New teaching model pays close attention not only to knowledge and phenomena but also to the essence and the design idea of the structure of human body. New teaching model attempts to arouse the students’strong desire for exploration and interests in medical sciences.

Objective:To construct recombinant adenovirus vector Ad5-CCL20,and detect the expression of CCL20 after Ad5-CCL20 transfected colon cancer cells CT-26.Methods:Genes encoding CCL20 was obtained from original plasmid double-digested with EcoR I/Sal I enzymes.The CCL20 DNA segments were linked into pDC316 to recombine shuttle plasmid pDC316-CCL20.After genome sequencing,we take shuttle plasmid pDC316-CCL20 and plasmid backbone pBHGIox_E1,3Cre co-transfecting 293T cells in mediation of liposome.The constructed recombinant adenovirus vector was named Ad5-CCL20.Lastly,after Ad5-CCL20 transfected CT-26 cells in vitro,the expression of CCL20 at different time points (12h,24h,36h and48h)was detected by Western blot and Elisa.Then,Culture supernatant was added into iDC and mDC to evaluate the chemotactic activity of CCL20.Results:The recombinant adenovirus Ad5-CCL20 were successfully constructed.The expression of CCL20 was detected by Western blot and Elisa.The level of CCL20 expression was increased with prolonged incubation of the infected CT-26 cells.Chemotaxis experiments show that the chemokine CCL20 had chemotactic activity to the iDC and mDC,but more obviouly for iDC (P＜0.05).Conclusion:The construction and obtain of recombinant adenovirus vector Ad5-CCL20 provide a new method for developing tumor immunotherapy.