Objective To investigate the effects of annonaceous acetogenin on proliferation and apoptosis in Raji cells and its mechanism.Methods Raji cells cultured in vitro were divided into control group,annonaceous acetogenin group and adriamycin group.Raji cells were effected by 6.25,12.5,25,50 ?g/mL annonaceous acetogenin.Proliferation of Raji cells were evaluated by MTT assays,apoptosis percentage was assessed by flow cytometry.Caspase-9 protein was detected by immunohistochemistry.Results The Raji cell proliferation rate of annonaceous acetogenin decreased compared with the control,that of 25,50 ?g/mL group were lower than adriamycin group,and it was related to the concentration,relying on the incubating time.The apoptosis rate was higher than control,that of 25,50 ?g/mL group were higher than adriamycin group,and it was related to the concentration and the incubating time.The expression of caspase-9 protein of annonaceous acetogenin group was higher than control,and it had a positively relationship with the concentration and incubating time.Conclusions Annonaceous acetogenin could inhibit cell proliferation in a dose-dependent and time-dependent manner in Raji cells,and it may induce Raji cells apoptosis by up-regulating caspase-9 expression.

The future war will inevitably to be in the land,the sea and the sky and so on hyperspace,and electromagnetic environment is very complex.How to eliminate interference of medical equipment under the complex electromagnetic environment,to promote the performance of the medical support,these is austerity and reality questions in current every level of medical and health organization.Only based on the existing,bold practice,independent innovation,the comprehensive medical support exercise under the complicated electromagnetic environment,to master various types of medical equipment in a complex electromagnetic environment of the characteristics and laws in order to give full play to existing health technologies and equipment performance,improve the timeliness of medical support.

BACKGROUND: Many experiments have proved that heavy-load movement training causes the acute increase of free radicals. The increase of endogenous free radicals and caused cellular and subcellular lipid peroxidation strengthening injure the structure and function of tissue cells, thereby, decrease motor ability. Sleep deprivation also causes the increase of oxygen free radicals.OBJECTIVE: To observe the changes in malondialdehyde (MDA) and glutathione (GSH) levels as well as superoxide dismutase (SOD) activity in serum of acute exhaustive exercise rats following different time periods of sleep deprivation.DESIGN: A randomized controlled animal experiment.SETTING: Laboratory of Exercise Sciences & Sports Medicine, Physical College, Hunan Normal University.MATERIALS: Thirty healthy male SD rats of clean grade, weighing about (220±13)g, provided by Experimental Animal Center of Hunan Agricultural University, were involved in this study.METHODS: This experiment was carried out in the Laboratory of Exercise Sciences & Sports Medicine, Physical College,Hunan Normal University from April 2006 to May 2006. Thirty rats were randomized into 5 groups: blank control group,simple exercise group, sleep deprivation 24 hours group, sleep deprivation 48 hours group and sleep deprivation 72 hours group, with 6 rats in each group; Rats in the blank control group were allowed to sleep normally, but not do exercise; Rats in the simple exercise group were allowed to sleep normally and executed after acute exhaustive exercise; Rats in the sleep deprivation 24, 48 and 72 hours groups were deprived their sleep for 24, 48 and 72 hours,respectively, then they were executed after acute exhaustive exercise. Method of gentle handling was used in creating rat models of sleep deprivation; Rats in the simple exercise group and sleep deprivation groups were forced to do exercise according to the rat exercise model project established by Bedford: treadmill gradient 10°, speed 19.3 m/min, all the exercise rats were exhaustive (Exhaustion criteria: At the end of exercise, rats reached 1/3 of runway over 3 times;Various stimulations for expelling were invalid. After running, rats presented with breathlessness, expression lassitude,ventral decubitus, slow stimulus response, weaker escape response in being captured). After experiment, rats in each group were executed under the anesthetic state, and their blood was taken out, and centrifuged after natural clotting.Supernatant fluid was taken for detecting MDA and GSH levels as well as SOD activity.MAIN OUTCOME MEASURES: Changes in MDA and GSH levels, and SOD activity in rat serum.RESULTS: Thirty rats were involved in the final analysis. ① MDA level in the simple exercise group was higher than that in the blank control group (P＜ 0.01). MDA level in the sleep deprivation 24 hours, 48 hours and 72 hours was higher than that in the simple exercise group, respectively (P ＜ 0.01). MDA level in the sleep deprivation 72 hours was statistically higher than that in the other sleep deprivation groups, respectively (P ＜ 0.01). ②GSH level in the simple exercise group was lower than that in the blank control group (P ＜ 0.01). GSH level in the sleep deprivation 24 hours group was higher than that in the simple exercise group [(P ＜ 0.01). GSH level in the sleep deprivation 24 hours and 48 hours groups was lower than that in the simple exercise group, respectively (P ＜ 0.05, 0.01). There were statistical differences in GSH level among sleep deprivation groups (all P＜ 0.01). ③ SOD activity in the simple exercise group was lower than that in the blank control group (P ＜ 0.01); SOD activity in the sleep deprivation 24, 48 and 72 hours groups was lower than that in the simple exercise group, respectively (P ＜ 0.01); SOD activity in the sleep deprivation 48 and 72 hours groups was significantly lower than that in the blank control group, respectively (P ＜ 0.01); There were statistical differences in SOD activity among sleep deprivation groups (P ＜ 0.01).CONCLUSION: Sleep deprivation can cause serumal oxidative stress injury of rats; With elongation of time of sleep deprivation and exhaustive exercise, oxygen free radical production in serum of rats accumulates more and more, ability to get rid of oxygen free radical becomes weaker and weaker, and injury to body is more and more obvious.Serumal oxidative stress status of acute exhaustive exercise rats following sleep deprivation

Objective:To analyze the characteristics of long non-coding RNA (lncRNA) expression in nonalcoholic fatty liver disease (NAFLD).Methods:lncRNA-mRNA microarray was conducted on the liver tissue samples from 10 patients with simple gallbladder stone (5 NAFLD liver samples and 5 normal liver samples),and the differentially expressed lncRNA was analyzed by bioinformatics technology.Results:Compared with the normal liver samples,there were abnormal expression of 1 735 lncRNAs and 1 485 mRNAs in NAFLD liver samples.Among them,535 lncRNAs and 760 mRNAs were up-regulated,1 200 lncRNAs and 725 mRNAs were down-regulated.Conclusion:Compared with normal liver,the expression oflncRNA in NAFLD tissues is obviously abnormal.These lncRNAs may play an important role in the occurrence and development of NAFLD.

Objective To investigate whether Th17/Treg imbalance existed,and whether VEGF165 attenuated the imbalance in allogeneic skeletal myoblasts transplantation(allo-SMT) for acute myocardial infarction (AMI).Methods There were 60 C57BL/6 male mice,3 to 4 weeks old,weighting 0.016-0.020 kg.On the day 1,2,4,and 7 after allo-SMT,the percentages of Th17 and Treg cells,and the ratios of Th17/Treg cells were analyzed by FCM in AMI group,AMI-S group(alloSMT) and AMI-Ⅴ group(with VEGF165 treatment).Subsequently,related proinflammatory and regulatory cytokines and key transcription factors ROR-γt mRNA and Foxp3 mRNA expression were examined by Bio-plex and RT-PCR respectively.Results On the day 1,2,4,and 7 after allo-SMT,the percentage of Treg,related cytokines concentrations and transcript factor Foxp3 mRNA in AMI-S group were lower than those in AMI group,while the AMI-Ⅴ group were higher than those in AMI groups.However,the percentage of Th17 cells,related cytokines concentrations,and ROR-γt mRNA in AMI-S group were higher than those in AMI group; but the AMI-Ⅴ group were lower than those in AMI group.Apparently,compared with AMI group,the ratios of Th17/Treg significantly ascended in AMI-S group and decended in AMI-Ⅴ group.Conclusion Th17/Treg imbalance participated in the formation and development of inflammation and immune response after allo-SMT.However,transfected VEGF165 could relieve the severity of Th17/Treg imbalance.

Objective To determine the normal range of hematological and visceral weight parameters of F 1 4-week and 8-week old, male and female transgenic CB6F1 mice.The influence of gender and week age on the biochemical parameters was assessed .Methods 4-week and 8-week old CB6F1 mice, half male and half female ( n=20 in each group) , were weighed alive , dissected to weigh the main viscera , and blood samples were collected to test the physiological and biochemical parameters .Results When 4-week old and 8-week old CB6F1 mice were compared , there were significant differences in 22 parameters (body weight, heart, liver, spleen, left ovary, left testis, right testis, WBC, RBC, HGB, HCT, MCV, MCH, PCT, MPV, PDW, LYM, TP, ALT, ALB, P and TG) (P<0.01 for all), and in 8 parameters (left kidney, right kidney, right adrenal, thymus, left ovary, RDW, MON%and BUN) (P<0.05 for all). When male and female 4-week CB6F1 mice were compared, there were significant differences in 14 parameters ( body weight, heart, liver, spleen, lung, left kidney, right kidney, MCHC, LYM, ALT, ALP, GLU, P and CHO) (P<0.01 for all), and in 6 parameters (right adrenal, WBC, PCT, MPV, TP and BUN) (P<0.05 for all).For male and female 8-week old CB6F1 mice, there were significant differences in 15 parameters (body weight, heart, liver, lung, left kidney, right kidney, MCV, PCT, LYM, LYM%, NEUT%, ALT, GLU, P and CHO) (P<0.01 for all), and in 5 parameters (WBC, RBC, MPV, NEUT and TP) (P<0.05 for all).Conclusions The normal range of hematological and visceral weight parameters of 4-week and 8-week old male and female CB6F1 mice are determined.Our study establishes normal detection indexes of CB6F1 mice and provides useful reference for its application .

Objective To improve the gene targeting efficiency with C57BL/6 embryonic stem ( ES) cells.Meth-ods Three different genetically modified C57BL/6 ES cell lines, named TLX3, Ai3K and SL, were microinjected into ICR, B6( Cg)-Tyrc-2J and BALB/c mouse blastocysts, respectively.The efficiency was statistically evaluated according to three aspects:blastocyst collection, chimera production and germline transmission.Results None of the three ES cell lines was germline transmitted with B6(Cg)-Tyrc-2J mice as blastocyst donors, while it was achieved with both BALB/c and ICR mouse blastocysts.Compared in the aspect of blastocysts collection, ICR mouse was much better than BALB/c mouse (P<0.05), and the chimera production efficiency of ICR mouse was comparable to that of BALB/c mouse (P =0.115). As to the germline transmission efficiency, that of BALB/c mice is significantly higher than that of the ICR mice ( P<0.01).Conclusions The germline transmission efficiency of BALB/c mouse is highest among these three mouse strains. However, it has the disadvantages of blastocyst collection, developmental delay and zona pellucida fragility, compared with ICR mouse.Therefore, ICR mouse is also a good candidate as blastocyst donor for embryonic stem cell microinjection.

BACKGROUND: Acute renal insufficiency (ARI) usually occurred following liver transplantation due to the surgical trauma and the application of immunosuppressant, which lack of unified diagnostic criteria. OBJECTIVE: To investigate the experience of diagnosis and treatment of ARI following liver transplantation.DESIGN, TIME AND SETTING: The experiment was performed at the 458 Hospital of Chinese PLA from January 2004 to December 2006.PARTICIPANTS: A total of 37 cases received liver transplantation, including 35 males and 2 females, aged 37-67 years, mean aged (48.5±8.9) years. All cases were divided into the liver cancer group (n=16) and liver cirrhosis group (n=21). The liver cirrhosis group included 16 cases with posthepatitic type B cirrhosis, 4 with posthepatitic type C cirrhosis, and 1 with alcoholic cirrhosis. All these cases were in decompensation stage. The final diagnosis was performed by pathological examination. METHODS: The removal of kidney and construction of blood outflow tract was achieved by modified piggy-back liver transplantation. The arterial blood gas analysis, blood routine examination, renal function and liver function were examined more than twice per day. The cephalosporins, Fluconazole and ganciclovir or vancomycin were used for 5-7 days to prevent infections.MAIN OUTCOME MEASURES: The incidence rate of acute ARI, clinical features and outcomes of patients were observed.RESULTS: ARI developed in 19 patients with liver transplantation, 5 patients died, 14 patients recovered in 2-3 weeks. The incidence of ARI following liver transplantation was associated with infection, bleeding shock, respiratory failure and acute respiratory distress syndrome (P < 0.05). CONCLUSION: The incidence of ARI following liver transplantation was 51.35%, with 26.32% mortality rate. The early diagnosis and treatment are the key steps for increasing successful rate of ARI treatment following liver transplantation.

Objective To investigate the effects of Qishen Erlian Decoction on serum MMP-1 and TIMP-1 levels in thioacetamide (TAA) induced liver fibrosis rats; To discuss its mechanism of action. Methods Liver fibrosis model was created by the TAA gavage method. 120 SD male rats were randomly assigned to control group, model group, colchicine group, Qishen Erlian Decoction low-, medium- and high-dose group (20 in each group). Each medication group was given relevant medicine for gavage. Control group and model group were given the same amount of normal saline for gavage, once a day for 5 weeks. HE staining and Masson trichrome staining were used to observe the pathological changes in liver tissue and liver tissue damage. Biochemistry, radioimmunoassay, and ELISA were used to detect the serum liver function, hepatic fibrosis index, MMP-1 and TIMP-1 levels. Results Compared with the model group, serum ALT, AST, TBIL, γ-GGT, HA, LN, Ⅳ-C and PCⅢ levels, MMP-1 and the ratio of MMP-1/TIMP-1 increased significantly and level of TIMP-1 decreased significantly in Qishen Erlian Decoction groups, with statistical significance (P<0.05, P<0.01). And there is a certain dose-effect relationship, with Qishen Erlian Decoction high-dose group the best effect. Conclusion Qishen Erlian Decoction can improve the liver function and liver fibrosis indexes, regulate the level of MMP-1 and TIMP-1, and prevent the progression of liver fibrosis.

Objective To establish a simple , fast and economic total DNA extraction method to serve the rapid identification of model mouse genotype in large number of mice .Methods Three methods, i.e.phenol extraction, isopropyl alcohol precipitation and mouse ear boiling methods were used to extract the total DNA from ten C 57-rasmodel mice.The purity and yield of DNA obtained by the three methods were compared , and polymerase chain reaction ( PCR) assay was used to compare the efficacy of the three extraction methods .Results Among the three methods , phenol extraction was the best and isopropyl alcohol precipitation was the poorest in DNA yield .In terms of DNA purity , the phenol extraction was the best and the mouse ear boiling method was the poorest .All the three methods could be used to extract the total DNA from mice serving as template of PCR reaction for the mouse genotype identification .The time consumption of three methods are 12.5 hr ,13 hr and 0.18 hr.Mouse ear boiling method was significantly lower than that of the other two methods ( P <0.01 ) ,.The obtained total DNA can be stored at conventional -20℃for 7 days and 30 days later still can be used as a template for PCR reaction .Conclusions Among the three methods studied , the mouse ear boiling method is simple and with the lowest cost , so it is feasible for total DNA extraction in scaled genotyping experiments .