Life science is based on an experimental basis, so the basic experimental teaching for biological and medical graduates is very important. Targeted basic experimental teaching content can help biological and medical graduates lay a solid experiment foundation and promote them to fol-low the development of cutting-edge bioscience.The basic experimental teaching content for biological graduate is set up into four experimental categories uniformly: molecular biology, proteomics experi-ments, bioinformatics experiments and large equipment use. Each category is further set into three experimental projects: validation experiments, comprehensive experiments and independent research experiment. By increasing the degree of openness of large instruments and equipment, improving teaching methods and perfecting appraisal methods, ggraduates' experimental skills, research ability and innovation ability are trained, and the quality of experimental teaching has been improved.

ObjectiveTo investigate the risk factors of metabolic syndrome ( MS ) in obese subjects.Methods A seven-year follow-up study was conducted in 413 simple obese subjects and 196 subjects with normal body weight who were recruited from community residents during physical examination in 2000.There was a 7 years follow-up.Anthropometrics,blood pressure,lipid profile,fasting blood glucose,and 2 h blood glucose after glucose loading were measured.Endothelium-dependent dilatation (EDD) test was also performed.Results Among 553 of 609 subjects who were followed up in 2007,there were 381 simple obese subjects ( simple obese group) and 172 normal weight subjects( normal weight group).Seven-year cumulative incidence of MS was 35.17％ in simple obese group and 8.14％ in normal weight group.In simple obese group,subjects with MS showed greater or higher levels of waist circumference( WC ),waist-hip ratio ( WHR ),triglyceride ( TG ),fasting plasma glucose ( FPG ),fasting insulin (FINS),and homeostasis model assessment for insulin resistance index (HOMA-IR) (all P＜0.05 ),and also decreased EDD( P＜0.05 ) as compared with those without MS.WC,WHR,and FINS were higher( all P＜0.05 ) and EDD was lower( P＜0.05 ) in subjects with MS of normal weight group than those without MS.Logistic analysis showed that the male gender,WC,WHR,FPG,HOMA-IR,and EDD were major risk factors of MS.Conclusion Central obesity,insulin resistance,and endothelial dysfunction are important independent risk factors for development of MS.

Objective To investigate the relationship between Trp64Arg mutation in the β3-adrenergic receptor (β3-AR) gene and the incidence of hypertension in obese subjects. Methods A seven-year follow-up study was conducted in 377 simple obese subjects who had Trp64Arg mutation in the β3-AR gene and some clinical metabolic parameters, such as body mass index, waist circumference, waist-hip ratio, blood pressure, lipid profile, fasting blood glucose, fasting insulin and insulin resistance index (HOMA-IR) were measured in the year of 2000 and 2007. Results The results of follow-up indicated that the incidence of hypertension in Trp64Arg heterozygote subjects were higher than that in Trp64Trp homozygote subjects (52.7% ,69/131 vs 37.0% ,91/246,P < 0.01), the difference was only seen in male (59.5%, 50/84 vs 45.1%, 64/142, P < 0.05). The incidence of hypertension in both Trp64Trp homozygote and Trp64Arg heterozygote subjects were higher in obese male than those in obese female (P < 0.01 or <0.05). After seven years, the blood pressure increased (9.7 ± 4.3)/(5.4 ± 4.0) mm Hg(1 mm Hg = 0.133 kPa) in Trp64Trp homozygote,and (12.8 ± 5.2)/ (7.9 ± 4.7) mm Hg in Trp64Trp heterozygote subjects. Compared with that in Trp64Trp homozygote subjects, lipid profile, fasting insulin and HOMA-IR was increased significantly in Trp64Trp heterezygote subjects (P < 0.05). Logistic analysis showed that β3-AR gene mutation, male, central obesity and insulin resistance were associated with the incidence of hypertension in obese subjects. Conclusion The β3-AR gene Trp64Arg mutation is the independent risk factor in the incidence of hypertension in male.

Objective To investigate the relationship between polymorphism of interleukin-1β(IL-1β)-31,-511C/T and chronic obstructive pulmonary disease.Methods Two hundred and sixty patients with chronic obstructive pulmonary disease (COPD) were selected as our subjects who were hospitalized in General Hospital of Kailuan from January 2009 to June 2012.At same time,the 260 healthy controls were recruited in medical examine center.The data was collected by the physical examination and a unified questionnaire.Enzyme-linked immunosorbent assay (ELISA) was used to measure the level of IL-1β in serum.Polymerize chain reaction-Restriction fragment length polymorphism (PCR-RLFP) was used to detect the genotypes of IL-1β (-31,-511).Multivariate logistic regression was used to analyze the relations between risk factors with susceptibility of COPD.Results The level of serum IL-1β was (3.92 ± 0.42) μg/L in COPD group,higher than that in control group ((2.69 ± 0.11) μg/L,t =12.889,P ＜ 0.001).The frequency of genotype of IL-1β-31 site in COPD group were 20.8％ (54/260) for TT,58.1％ (151/260) for CT and 21.2％ (55/260) for CC respectively.Meanwhile IL-1β-31 genotype rate were 19.2％ (50/260),58.8％ (153/260) and 21.9％ (57/260) respectively in control group and no significant difference was found between two groups (x2 =0.203,P =0.904).The frequency of genotype of IL-1β-511 site were 20.0％ (52/260) for CC,63.1％ (164/260) for CT and 16.9％ (44/260) for TT in COPD group.The three genotype rate in control group were 21.9％ (57/260),60.8％ (158/260) and 17.3％ (45/260) respectively,and no significant difference was found between two groups (x2 =0.352,P =0.838).Moreover there was also no significant difference in terms of gender(P ＞ 0.05)Conclusion The concentration of IL-1β in serum in COPD group was higher than in control group.The polymorphism of-31 and-511 were proved non association with COPD.

Exercise is one of the important techniques of cardiovascular rehabilitation. Exercise can reduce inflammatory response to improve endothelial function, and improve mitochondrial function to increase myocardial cell activity. For cardiovascular risk factors, exer-cise can promote the activity of lipoprotein, increase the level of high-density lipoprotein;improve the function of insulin receptor to reduce insulin resistance, reduce platelet aggregation and improve endothelial function to reduce blood pressure. For the respiratory system, aerobic exercise can improve the function of respiratory muscle, thus relieve the dyspnea. Exercise can promote the activation of immune factor and increase metabolism, to increase immune function and anti-aging. Resistance exercise can improve mitochondrial function and promote fi-ber type conversion, to improve the function of skeletal muscle system.

Objectives To observe the effect of Luoyutong capsule on neurological function following focal cerebral ischemia-reperfusion in rats and to preliminarily study the protective mechanism of Luoyutong capsule for focal cerebral ischemia-reperfusion in rats. Methods A rat model of middle cerebral artery occlusion (MCAO)was induced by the modified Longa method. After 1. 5 h of ischemia,reperfusion started. Ten male SD rats were selected as sham operation group,and forty male SD rats were randomly divided into 4 groups:Model (MCAO),Luoyutong moderate-dose (LYTM),Luoyutong high-dose (LYTH),and citicoline sodium (CS)groups (n=10 in each group). At day 3 and 7 after modeling,the neurological function of the rats was evaluated by using 12 neurological score and forelimb placing test. Western blotting was used to detect the expression levels of brain derived neurotrophic factor (BDNF),basic fibroblast growth factor (b-FGF),and phosphor/protein kinase (p-AKT/AKT)on the ischemic side of the rats and in the ipsilateral brain tissue at day 3 after modeling,as well as the expression level of Caspase-12 at day 7 after modeling in the ipsilateral brain tissue,and a comparison was performed among the groups. Results (1 )Neurological score:At day 3 after modeling,there was no significant difference between the 12 neurological score and the forelimb placing test score (all P>0. 05);At day 7 after modeling, there were obvious improvement in the LYTM,LYTH,and CS groups compared with model group (all P<0.05). (2)The results of western blot showed that①compare with the sham operation group,the expression levels of BDNF and b-FGF were reduced obviously (all P<0.05);compare with the MCAO group,the expression levels of the LYTM,LYTH and CS groups could be up-regulated,particularly in the LYTH group (P<0. 01);② compare with the sham operation group,the expression level of p-AKT/AKT in MCAO group was decreased obviously (P<0. 05);compare with the MCAO group,the expression levels of p-AKT/AKT of the LYTM,LYTH,and CS groups were increased,particularly in the LYTH and CS groups (all P<0. 05);③ compared with the sham operation group,the expression of cleavage Caspase-12 was increased obviously in the MCAO group (P<0. 05). Compared with the MCAO group,the expression levels of proCaspase-12 and cleavage Caspase-12 had a decreasing trend in the LYTM and LYTH groups,but there were no significant differences (all P >0. 05);the expression levels of proCaspase-12 and cleavage Caspase-12 in the CS group were obviously lower than those of the MCAO group (P<0. 05). Conclusion Luoyutong capsule may play a protective effect for focal cerebral ischemia-reperfusion injury in rats by promoting neural survival and regeneration,and this protective effect may be associated with the inhibition of neuronal apoptosis.

Objective To investigate the relationship between Trp64Arg mutation in β3-adrenerglc receptor (β3-AR) gene and the incidence of metabolic syndrome (MS). Methods A seven-year follow-up study was conducted in 386 simple obese subjects and 175 normal weight subjects in whom geno-typing of Trp64Arg mutation in β3-AR gene was examined in 2000. Results There were no differences between a Trp64Trp homozygote group and a Trp64Arg heterozygote group of whether obese or normal weight subjects with respect to adiposity, blood pressure, lipid profile, fasting blood glucose and fasting insulin in the baseline. The results of follow-up indicated that the incidence of MS in the Trp64Arg heterozygote group was higher than that in the Trp64Trp homozygote group of obese males (54. 76% vs 40. 85% ,P <0. 05) but not in the group of obese females. The incidences of MS both in the Trp64Trp homozygote group and Trp64Arg heterozygote group were higher in obese males than in obese females (40. 85% vs 18. 27% and 54. 76% vs 21.28% ,all P <0. 01 ) . No significant differences were found in incidences of MS both in the Trp64Trp homozygote group and Trp64Arg heterozygote group of normal weight subjects whether the comparison was made between males and females respectively or between males and females. The overall incidence of MS in the obese subjects were significantly increased than that in the normal weight subjects whether there was genevariant or not(31.30% vs 6. 03% and 42. 75% vs 12. 73%, all P <0. 01 ). Logistic analysis showed thatβ3-AR gene variant was associated with increased incidence of MS in males. Conclusion β3-AR gene Trp64Arg mutation is an independent risk factor for the incidence of MS in males.Conclusion β3-AR gene Trp64Arg mutation is an independent risk factor for the incidence of MS in males.

Objective To observe the protein and mRNA expression of LOX-1,eNOS and PPARγ in type 2 diabetic rat aorta,and to investigate the effect of rosiglitazone intervention.Methods Totally 80 Wistar rats (7-week-old male) were randomized into the control group,high fat diet group,diabetic group,and rosiglitazone treatment group (n=20 each).Type 2 diabetes model was developed by intraperitoneal injection with a low dose of streptozotocin,and rats in rosiglization treatment group were treated with rosiglitazone by intragastric administration.After treatment with rosiglitazone for 6 and 12 weeks,animals were sacrificed.Aorta were collected for detecting the protein and mRNA expressions of LOX-1,eNOS and PPARγ,and the differences in expression levels were compared among groups.Results After 6 and 12 weeks,the protein expressions of LOX-1 were up-regulated in diabetic group and rosiglitazone treatment group as compared with control group and high fat diet group (all P＜ 0.01).The protein expression of LOX-1 was down-regulated in rosiglitazone treatment group as compared with diabetic group (P ＜ 0.05).The aorta protein expressions of LOX-1 in high diet group,diabetes group and rosiglitazone treatment group were upregulated after 12 weeks as compared with 6 weeks (all P＜0.01).After 6 and 12 weeks,the aorta protein expressions of eNOS were down-regulated and PPARγ were up-regulated in high fat diet group,diabetic group and rosiglitazone treatment group as compared with control group (all P＜0.01)The aorta protein expression of eNOS was down-regulated and PPARγwas up-regulated in diabetes group as compared with high fat diet group and rosiglitazone treatment group (all P＜0.01).The aorta protein expressions of eNOS in diabetes group and rosiglitazone treatment group were downregulated after 12 weeks as compared with 6 weeks (all P＜0.01).After 6 and 12 weeks,the aorta mRNA expressions of LOX-1 and PPARγ were up-regulated,but the mRNA expressions of eNOS were down-regulated in high fat diet group,diabetes group and rosiglitazone treatment group as compared with control group (all P＜0.05).The aorta mRNA expressions of LOX-1 and PPARγ were up-regulated,but the mRNA expressions of eNOS were down-regulated in diabetes group and rosiglitazone treatment group as compared with high fat diet group (all P＜0.05).The aorta mRNA expressions of LOX-1 and PPARγ were down-regulated,but the mRNA expressions of eNOS were upregulated in rosiglitazone treatment group as compared to diabetic group (all P＜0.01).Conclusions Both hyperglycemia and hyper-lipoproteinemia can induce early coronary atherosclerosis in rats with the abnormal protein and mRNA expressions of LOX-1,eNOS and PPARγ in rat aorta,and the abnormal expressions are more obvious in hyperglycemia combined with hyperlipoproteinemia.Thiazolidinediones can reverse the above abnormal expressions in diabetic rats.

Objective To analyze the relationship between the expression of MHC class I chain-related gene A/B(MICA/B) on different human cancer cells and their sensitivity to NK cell cytotoxicity.Methods Hie expression MICA/B in K562,Bcap37,769P and A549 cells was measured by FACS.Isolation of NK cells were obtained by the modified RosetteSep~((R)) procedure.Cytotoxicity of NK cells at different ef0.50)%,(90.72±0.64)%,(55.59±0.55)%,and(3.84±0.10)% respectively.The purity of NK cells obtained by the modified RosetteSep~((R)) procedure was(96.52±2.42)%,whereas the cytotoxicity of NK cells against K562,Bcap39 and 769P was much higher than that of A549(P<0.01).The expression of MICA/B was significantly correlated with the cytotoxicity of NK cells at E:T ratios of 5:1,10:1 and 20:1 respectively(P<0.01).Conclusion The MICA/B expression on cancer cell lines was correlated with NK cell-mediated cytolysis and influenced the cytotoxicity of NK cells.

Objective To extract exosomes from dendritic cell (DC)and investigate antitumor effect of the special cytotoxic T lymphocytes activated by exosomes against 769-P.Methods Monocytes from peripheral blood of healthy donors were cultured to induce DC in intro and their phenotypes were analyzed by FACS.The exosomes were extracted from DC by ultrafiltration and ultracentrifugation and observed under electric microscope.Proliferation of T cells and the cytotoxicities of CTL against 769-P induced by exosome were measured by MTT.Results The exosomes could be separated from culture supernatant of DC by ultrafiltration and ultracentrifugation.The combination of the exosomes from DC and the 769-P antigen could effectively stimulate T cell proliferation and enhance their cytotoxicity, the absorption value of T cell proliferation(1.68±0.22), the cytotoxicity of CTL(38.23±2.16)%.Conclusion The exosomes extracted by ultrafiltration and ultracentrifugation can present tumor antigen to T cells and induce the cytotoxicity of CTL.