Objective:To investigate the influence of combined application of histone deacetylase inhibitor(HDACi)MS-275 and 5-FU on the apoptosis and cycle arrest on human hepatoma cell line HepG2 and unclose the related mechanism.Methods:Divided all the cells into 4 groups:control group,MS-275 group,5-FU group and drug combination group.Flow cytometry(FCM)was used to examine the effects of 5-FU with MS-275 on the apoptosis and cell cycle of HepG2 cells.Bcl-2,Bax,CyclinD1 and P21 protein were determined by Western blot assay.Results:5-FU and MS-275 combination could inhibit HepG2 cell growth through G0-G1 arrest,and induce apoptosis,both time and dose dependently.The combination of two agents increased P21 protein levels and decreased Bcl-2,CyclinD1 protein levels.The levels of Bax protein were not changed.Conclusion:The combination of 5-FU and MS-275 can induce apoptosis and cell cycle arrest,the effect might be associated with down-regulating expression of Cyclin D1 and Bcl-2 protein and upregulating the expression of P21 protein.

Vascular smooth muscle cell (VSMC) proliferation and migation play important roles in the occurrence and development of atherosclerosis and restenosis after intravascular stenting.The studies in recent years have shown that kallikrein-kinin system (KKS) is closely correlated with VSMC proliferation and migration in cytokines and transduction pathways.Therefore,investigating the roles of KKS in the VSMC proliferation and migation process has great significance in clinical prevention and treatment of atherosclerosis and restenosis after intravascular stenting.

Objective To explore the extent of lung injury induced by hyperoxia,and the activity of ubiquitin-proteasome pathway(UPP) in pathophysiological progress of lung tissue in early stages.Methods Adopted completely random design,20 SD rats were divided into hyperoxia group and air control group.For the air control group,the oxygen concentration exiting the cages was analyzed with oxygen monitor and oxygen concentration remained at 210 mL/L for 72 hours;while in the hyperoxia group,the condition changed into high-density oxygen(950 mL/L) for 72 hours to estimate the hyperoxia lung injury in rats model.The contents linked morphology as pathological classification in gross finding,pathological score of lung injury and the index of pneumonedema-the ratio of moist to dry weight of lungs were mea-sured.The expressions of ubiquitin protein and the activity of proteasome 20 S and the active statement of ubiquitin-proteasome pathway were detected by immunohistochemistry and Western blot methods.Results 1.The hyperoxia lung injury rat model was successfully duplicated.2.In hyperoxia group,pulmonary edema with increased ratio of moist to dry weight of lungs could be found(P=0).3.Macroscopic observation: bright red and full-stacked lung tissue,foliated or local hemorrhage on the surface,but little pleural effusion was observed in hyperoxia group.There was statistical significance of pathological classification in gross finding between hyperoxia group and air control group(P=0.005).Light microscope observation:swelled alveolar epithelium,widened alveoli wall,capillary engorgement and telangiectasis,obvious edema in interstitial tissue of pulmonary aveolus and alveolar space,increased inflammatory cells were observed in hyperoxia group.The findings of pathological score of lung injury indicated more serious injure than control group(P=0).4.The increased expression of ubiquitin protein in lung tissue was discoved by using immunohistochemistry and Western blot findings after hyperoxia exposure 72 hours.(P=0).5.The acti-vity of proteasomes 20 S in hyperoxia group was higher than that in control group(P=0).Conclusions The mainly pathological changes of lung are generated through hyperoxic exposure for 72 hours,including alveolar epithelial cell and vascular endothelial cell injury diffusely,inflammatory cell infiltration and pulmonary edema.Active the ubiquitin-proteasome pathway is connected with the pathophysiological process of lung injury in the initial stages of hyperoxia-exposure.

Objective To investigate the protective effects of the ubiquitin proteasome inhibitor MG-132 on p38 signaling pathway and apoptosis in lung injury induced by hyperoxia. Methods Twenty-six SD rats were randomly divided into 4 groups: normal control group(n=5),MG-132 control group(n=5),hyperoxia group(n=8) and MG-132 hyperoxia group(n=8).Hyperoxia lung injury rat models were established,and proteasome inhibitor(0.5 mg/kg) was intraperitoneally injected in control group and MG-132 hyperoxia group once daily.The resected lungs were histopathologically examined,and cell apoptosis and expression of ubiquitin and p38 were detected by TUNEL and immunohistochemistry,respectively.Results After hyperoxia exposure,there were edema and inflammatory cell infiltration in the lung tissues of SD rats.The apoptosis index and expression of p38MAPK of hyperoxia group were higher than those of normal control group and MG-132 hyperoxia group(P

Objective To evaluate the value of a CT ribs unfolding algorithm in the diagnosis of rib fracture. Methods Retrospective analysis of 180 patients who suffered chest trauma to do the chest or/and abdominal CT examination, and they also had the surgical or CT referral information. The images of these patients were postprocessed by software(Bone Reading software)and hand-drawn method(multi-point hand-painted CPR method). The rib fracture was observed and the time of reading was record. The diagnosis of fractures was confirmed by follow-up review or surgery. The fractures diagnosis sensitivity of the two post-treatment methods were measured, and the McNemar test was used to compare the difference between the software method and the hand-drawn method. Results Eight patients were excluded due to program failure, 172 cases were included in the study. Of the 172 patients, 63 patients suffered 259 fractures(178 ribs). The sensitivity of the software group was 91.7％(475/518), which was higher than that of the hand-painted group(86.3％, 447/518), the difference was statistically significant(P=0.005). The time of reading were (30.3 ± 3.3)and(173.2 ± 4.5)s, respectively, and the difference had statistically significant(P=0.001). Conclusion Compared to the traditional CPR method, the bone reading technique was used in patients with rib fractures during thoracic CT postprocessing can shorten the reading time and increase the sensitivity of the diagnosis.

The optimized cultural medium of fermentation for Candida sp.mutant strain YQ5 to produce S-adenosylmethionine was studied.The results of single factor experiment showed that the most favorable pH value,carbon source,nitrogen source organic nutrient is 6.0,8% sucrose,1.5% tryptone and 2% yeast extract,respectively.As to inorganic salts,MgSO4?7H2O,CaCl2,FeSO4?7H2O,CoCl2,CuSO4?5H2O,H3BO3 could improve accumulation of the intercellular SAM.The ingredients of the culture medium are also opti-mized by the orthogonal experiment.Fermentation for 48 h under the optimal condition resulted in AdoMet production at 1740.0 mg/L.

OBJECTIVETo observe the effects of proteasome inhibitor MG-132 on hyperoxic lung injury in rats and explore the mechanism.

METHODSThirty SD rats were randomly divided into 3 groups, namely the normoxic group, hyperoxic group, and hyperoxic with MG-132 treatment group, and rat models of hyperoxic exposure-induced lung injury were established in the latter two groups. After pathological grading of the lung injury under optical microscope and determination of the wet/dry weight ratio of the lung tissue, the expressions of ubiquitin protein and nuclear factor-kappaB (NF-kappaB) p56 and the activity of proteasome 20S and myeloperoxidase (MPO) were detected. Tumor necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6) expressions in the lung tissue were also detected.

RESULTSThe rats with hyperoxic exposure showed obvious pulmonary edema and increased wet/dry weight ratio of the lung tissue (P<0.01), which were significantly alleviated with MG-132 treatment (P<0.01). Compared with the normoxic group, hyperoxic exposure resulted in significant lung pathologies (P<0.01), which was reduced after MG-132 treatment. Immunohistochemistry and Western blotting demonstrated increased expression of ubiquitin protein in the lung tissue after hyperoxic exposure (P<0.01), which was lowered by MG-132 treatment (P<0.01). Proteasome 20S activity was obviously enhanced in the hyperoxic group (P<0.01) but lowered by MG-132 treatment (P<0.01). Hyperoxic exposure also caused obviously enhanced MPO activity and expressions of NF-kappaB, TNF-alpha, and IL-6 (P<0.01), which were all reduced by MG-132 treatment (P<0.05).

CONCLUSIONMG-132 alleviates hyperoxic lung injury probably by inhibiting the NF-kappaB/inflammatory factor pathways.

Animals ; Animals, Newborn ; Cysteine Proteinase Inhibitors ; pharmacology ; Female ; Hyperoxia ; complications ; Interleukin-6 ; metabolism ; Leupeptins ; pharmacology ; Lung Injury ; etiology ; metabolism ; pathology ; Male ; NF-kappa B ; metabolism ; Peroxidase ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Tumor Necrosis Factor-alpha ; metabolism ; Ubiquitin ; metabolism

ONYX-015 and H101 are E1B 55-kDa protein-deficient replicating C group adenoviruses that are currently in clinical trials as antitumor agents. However, their application in cancer gene therapy is limited by the native tropism of C group adenoviruses. This is in part due to low expression of the C group adenovirus receptor (coxsackievirus-adenovirus receptor, CAR) on malignant tumors. An H101-based chimeric virus vector containing sequences encoding the Ad35 fiber domain instead of the Ad5 fiber (H101-F35) was constructed. This modification allowed infection of tumor cells through CD46, a membrane protein over-expressed on tumors. The CAR and CD46 RNA expression was evaluated by RT-PCR method. H101-F35 conferred a stronger cytocidal effect than H101 and ONYX-015 in tumor cell lines that lacked CAR expression (MDA-MB-435 and MCF-7), while the cytocidal effect of H101-35, H101 and ONYX-015 was similar in high-level CAR expressing cancer cell lines (A549, NCI-H446, Hep3B, LNCaP, ZR-75-30 and Bcap-37). In an MDA-MB-435 xenograft mouse tumor model, tumor growth in mice receiving H101-F35 was significantly inhibited compared with mice injected with H101. These results suggest that the chimeric oncolytic adenovirus H101-F35 vector might be a useful candidate for gene therapy of cancer.

The study was purposed to explore the effect of panel reactive antibody (PRA) serum from patients with β-thalassemia on proliferation and apoptosis of the CD34(+)cells from cord blood and its mechanism. CD34(+) cells of umbilical cord blood were incubated with different sera and complement respectively. After incubation, the samples were centrifuged and the supernatants were collected for lactate dehydrogenase (LDH) detection, and the CD34(+) cells were harvested and measured for the apoptosis by flow cytometry with Annexin V/PI. The intracellular DNA synthesis were also quantified by [(3)H]TdR incorporation using liquid scintillation counter. The results showed that concentration of LDH in PRA positive groups was higher as compared with control group, and the DNA synthesis of CD34(+) cells in PRA positive groups were inhibited. There were no differences in the percentage of cell apoptosis and necrosis among different groups. It is concluded that thalassemic serum PRA impairs the cell membrane, inhibits the DNA synthesis, which can be increased by addition of the complement, but PRA had no significant effect on apoptosis of CD34(+) cells.
Antibodies ; blood ; immunology ; Antigens, CD34 ; Apoptosis ; immunology ; Cell Proliferation ; Child ; Fetal Blood ; cytology ; Humans ; beta-Thalassemia ; blood ; immunology ; pathology