The teaching method, teaching process and teaching model of programming course were reformed in Beijing University of Traditional Chinese Medicine by combining task-driven and flipped classroom teaching con-cept with BB network platform to provide a free and open environment for students in order to train their capability of analyzing and solving problems and the skills of independent learning and life-long learning. Good results were achieved in two rounds of task-driven plus flipped classroom teaching, which are of significance for the teaching model and teaching structure reform of other courses.

Objective To investigate the relationship between the serum apolipoprotein B (ApoB)/ ApoA-Ⅰ ratio and intracranial artery stenosis in young patients with ischemic stroke.Methods Patients with ischemic stroke aged from 18 to 45 were enrolled in the study.Brain CT angiography was used to evaluate the degree of intracranial artery stenosis,and the concentrations of serum total cholesterol,triglyceride,highdensity lipoprotein cholesterol,low-density lipoprotein cholesterol,ApoA-Ⅰ,and ApoB were detected.The ratio of ApoB/ApoA-Ⅰ was calculated.The Demographic and clinical characteristics of the intracranial artery stenosis group and the non-intracranial artery stenosis group were compared.Multivariate logistic regression analysis was used to identify the independent risk factors for intracranial artery stenosis in young patients with ischemic stroke.Results A total of 161 young patients with ischemic stroke were enrolled,including 89 in the intracranial artery stenosis group and 72 in the non-intracranial artery stenosis group.The constituent ratios of diabetes mellitus (20.2％ vs.6.9％;x2 =4.641,P =0.032),smoking (47.5％ vs.15.2％;x2 =15.121,P=0.001),hyperlipidermia (56.1％ vs.48.6％;x2 =4.197,P=0.040),as well as the radios in serum high-density lipoprotein cholesterol (1.29 ± 0.30 mmol/L vs.1.65 ± 0.34 mmol/L;t =7.131,P=0.002),ApoA-Ⅰ (1.49 ± 0.65 g/L vs.1.63 ± 0.23 g/L;t =2.751,P =0.001),ApoB (1.49 ± 0.65 g/L vs.1.63±0.23 g/L;t=2.751,P=0.001),and ApoB/ApoA-Ⅰ ratio (1.49±0.65 vs.1.63± 0.23;t =2.751,P=0.001) had significant differences between the two groups.Multivariate logistic regression analysis showed that diabetes (odds ratio [OR] 3.052,95％ confidence interval [CI] 1.186-7.856;P =0.021),smoking (OR 2.997,95％ Cl 1.456-6.172;P =0.003),hyperlipidemia (OR 4.745,95％ CI 2.108-10.668;P =0.001),ApoB (OR 4.861,95％ CI 3.029-7.802;P=0.001),and ApoB/ ApoA-Ⅰ ratio (OR 5.684,95％ CI 2.215-14.584;P=0.002) were the independent risk factors for intracranial artery stenosis in young patients with ischemic stroke,while HDL-C (OR 0.561,95％ CI 0.354-0.888;P=0.014) and ApoA-Ⅰ (OR 0.065,95％ CI 0.010-0.409;P=0.004) were the independent protective factors.After adjustment for hypertension,diabetes,smoking,hyperlipidemia,HDL-C,ApoA-Ⅰ,and ApoB,ApoB/ApoA-Ⅰ ratio was still an independent risk factor for intracranial artery stenosis in young patients with ischemic stroke (each increase of 1 standard deviation,OR 4.255,95％ CI 2.348-7.711;P=0.001).Conclusion ApoB/ApoA-Ⅰ ratio is an independent risk factor for intracranial artery stenosis in young patients with ischemic stroke.

Objective:To remove murine embryonic stem cells(mESC)from the differentiating cell culture and purify the differentiated cells by Magnetic Activated Cell Sorting(MACS).Methods:Neural differentiation of mESC was induced by a 5-stage method.The specific cell surface marker,SSEA-1,was used to identify ES cells in the differentiating cells.The optimal dilutions of mouse anti mouse SSEA-1 IgM primary antibody and FITC conjugated goat anti mouse secondary antibody were determined before the flow cytometry test.The incubation time and incubation temperature of primary antibody were all optimized to make the cytometry test accurate.After the optimization,stage 4 cells were dissociated into single cell suspension,incubated with antibody of SSEA-1 and microbeads conjugated goat anti mouse IgM,and then sorted through the magnetic field.The rate of SSEA-1 positive cells in pre-and post-separation groups was assessed by flow cytometry,and the viability of cells was evaluated by trypan blue staining counting under light microscopy.Results:The proportion of SSEA-1 positive cells in the separated cells can be reduced from(7.19?1.36)% to(1.34?0.80)%.The survival rate of sorted cells was more than 92%,similar to that of pre-separation cells.Conclusions:The MACS system we used can effectively remove mESC from the differentiated cells.The sorted cells will be well provided for the subsequent studies about transplantation therapy.

OBJECTIVETo investigate the cytotoxicity of bromoxynil on SH-SY5Y cells and its effect on the expression of nuclear factor-kappa B (NF-κB) and I kappa B alpha (IκBα) in SH-SY5Y cells.

METHODSSH-SY5Y cells were exposed to bromoxynil (10, 50, or 100 µmol/L) for 24 and 48 h, and other SH-SY5Y cells, which were used as a control, were exposed only to dimethyl sulfoxide. After 24 and 48 h of exposure, the morphological changes of these cells were observed under an inverted microscope, and the cytotoxicity of bromoxynil was measured by MTT assay. The cellular proliferation was examined by cell counting after 12, 24, 48, 72, and 96 h of exposure. After 24 h of exposure, the expression of NF-κB was evaluated by Western blot and immunocytochemistry, and the expression of IκBα was evaluated by Western blot.

RESULTSThe cellular proliferation inhibition rates (CPIRs) of 50 and 100 µmol/L groups were significantly higher than that of the control group after 24 and 48 h of exposure (P < 0.05); the CPIR was significantly higher after 48 h than after 24 h in the two groups (P < 0.05). The growth curve revealed that these groups began to show differences in cell count at the 24th of exposure and that the differences were even more marked as the exposure went on (F = 17.15, P < 0.05). The control group had a significantly increased cell count at the 48th, 72nd, and 96th h of exposure (P < 0.05); the 10 and 50 µmol/L groups had a significantly increased cell count at the 72nd and 96th h of exposure (P < 0.05); the 100 µmol/L group showed no significant change in cell count during 96h of exposure. The 50 and 100 µmol/L groups hada significantly longer cell doubling time than the control group (P < 0.05). The immunocytochemistry showed that as the dose of bromoxynil increased, the brownish yellow particles in the cytoplasm and nuclei became darker, the expression of NF-κB was upregulated, and the nuclear translocation of NF-κB was increased. The Western blot showed that the 100 µmol/L group had significantly higher expression of NF-κB in the nuclei than the control group (P < 0.05) and that the 50 and 100 µmol/L groups had significantly lower expression of IκBα in total proteins than the control group (P < 0.05).

CONCLUSIONBromoxynil can inhibit the proliferation of SH-SY5Y cells under this experimental condition, which may be related to activation of NF-κB.

Cell Line, Tumor ; Cell Proliferation ; drug effects ; Humans ; I-kappa B Proteins ; metabolism ; NF-KappaB Inhibitor alpha ; NF-kappa B ; metabolism ; Nitriles ; toxicity

Adult ; Aged ; Electroacupuncture ; Female ; Humans ; Male ; Middle Aged ; Muscular Atrophy ; therapy ; Spondylosis ; therapy ; Treatment Outcome

Objective To detection the urine of bacteria hyphae and intracellular bacterial communities in patients with indwelling urinary catheter and discuss intracellular bacterial comnmunities in the pathogenesis of catheter-related urinary tract infection.Methods From May 2014 to February 2016,95 cases with D-J stent indwelling were enrolled in this study,including 38 male patients and 57 female patients.The mean age was (43 ±21)years old,ranging from 25 to 83 years old.We recorded those patient g clinical symptoms,middle urine culture results.If the middle urine culture was positive,further pathology test and scanning electron microscopy for bacteria hyphae and intracellular bacterial communities would be considered.Results The middle urine culture showed positive in 21 cases (22％,21/95);The classification of bacteria included E.coli in 11 cases,dung enterococcus in 2 cases,klebsiella pneumonia in 4 cases,pseudomonas aeruginosa in 3 cases,epidermis staphylococcus aureus in 1 case.Among those 21 patients,9 cases had the symptoms of fever and shiver.Urine pathology testing found hyphae in 6 cases (6％,6/95).all others were E.coli infection.For scanning electron microscope,6 cases were found rodshaped bacteria and hyphae.3 cases were found intracellular bacterial communities.Conclusions The presence of intracellular bacterial communities made urothelial itself the source of endogenous bacteria of urinary tract infection.Catheter-related urinary tract infections in patients with recurrence maybe basically homology bacteria.

Objective To explore the possibility of the inner vertical outer spiral complex tubular urethra scaffold vascularization in repairing long segment of urethral defect.Methods From August 2014 to October 2015,27 clean male New Zealand white rabbits were divided into 3 groups,S1 group was transfected recombinant vascular endothelial growth factor(VEGF) gene lentiviral vector group.S2 group was vascular pedicle transfer tube group.C group was simple stent group.A 3.0 cm inner vertical outer spiral complex scaffold was constructed by using the small intestine acellular matrix (SIS) and polylactic acid copolymer (PLGA) modified by type Ⅰ collagen surface,and adipose-derived mesenchymal stem cells (ADSC) and smooth muscle cells after transformation from New Zealand white rabbits.In S1 group,the seed cells were transfected by recombinant vascular endothelial growth factor (VEGF) gene lentivirus,which express VEGF protein.The complex scaffold was used to repair 3.0 cm rabbit urethral defect In S2 group,the untransfected cells were seeded into the scaffold and embedded in the skin near the groin artery 3 weeks for repairing urethral defect with vascular pedicle transfer tube.In group C,the unseeded scaffold was used to repair the urethral defect alone.Postoperative observation and urethrography were followed 4,8 and 24 weeks after implantation.The HE staining,fluorescence tracing,immunohistochemical and scanning electron microscopy were evaluated at the same phase.Results In S1 group,there were one urinary fistula and one urethral stricture-related death,respectively.The urethra was smooth and patent,histological examination showed active hyperplasia of urethral capillary.In S2 group,there were one urinary fistula and two urethral stricture-related deaths,respectively.The urethral was rough,local thinning or dilated.Fat accumulation and mucosal contraction were found in the urethral submucosal,respctively.In C group,there were one urinary fistula,three hypospadias,and three urethral stricture-related deaths.The thickness of the urethra was uneven and stricture bending.The urethral mucosa was poorly repaired and the scar was narrow.HE and CD31 staining showed that S1 and S2 groups were active in the proliferation of urethral capillaries,and the angiogenesis was abundant.VEGF staining showed that the cytoplasm of endothelial cell layer,smooth muscle layer of vascular wall and the urothelial epithelial cell layer were fully expressed at 24 weeks,especially in epithelial cell layer.CKpan staining showed that the epithelium of S1 and S2 group developed to stratified epithelium,and the morphology of urethra was similar to normal urethra at 24 weeks.The urethral epithelial in C group of grew poor as single-level,irregular arrangement,24 weeks is still a lack of effective stratified epithelium.HE and oα-SMA staining showed that the smooth muscle and actin gradually increased in group S1 and S2,α-SMA staining in group C was scarce and increased at 24 weeks.PLGA was encapsulated by the surrounding tissue and the structure of electrospinning was clear after 4 weeks,absorbed and degraded after 8 weeks and absorbed after 24 weeks.Conclusions The inner vertical outer spiral comnplex tubular urethra scaffold maybe a reasonable method in repairing long segment urethral defects,and the methods of tubular urethra scaffold vascularization by transfected VEGF gene recombinant lentiviral vector and vascular flap deserve more research.

To investigate the expression of alpha-smooth muscle action (alpha-SMA) in the renal tubulointerstitium in patients with kidney collateral stasis.

In this study,nearly 200 endophytic bacteria were isolated from different part of Huperzia serrata, over 60 bacterium with clear antifungal activity were selected from those cultures.Among them,strain H-6 exhibited the highest antifungal activity which was strongly inhibits the growth of many plant pathogenic fungi such as Sclerotinia scleroliorum,Fusarium graminearumt,Sclerotinia libertiana,Phytophthora capsici Leonia and Sesame fusarium wilt.According to the characteristics of morphology,physiology and biochem- istry tests and the comparison of 16S rDNA sequence,the strain H-6 was similar to the Burkholderia.So strain H-6 was identified as Burkholderia sp.H-6.The results also showed that Burkholderia sp.H-6 was markedly different from Burkholderia cepacia that was applied widely in agriculture as antagonistic bacteria. The medium and culture conditions of the strain all were optimized by single factor and orthogonal experiments.In the investigation of the culture condition,growth was carried out in a basal medium(potato juice)and gradually supplemented with the various ingredients to be investigated.The major ingredients be- ing investigated included carbon sources and nitrogen sources.The optimal antifungal activity production condition is growth in a medium(potato juice with 2.5%mannitol and 0.1%NaNO3),initial pH 4.0 at 28℃.

The type of basic media and the contents of plant growth substances were investigated by orthogonal design experiment,and also the effects of different culture conditions on the growth of suspension cells and the accumulation of total flavonoids in Eucommia ulmoides were studied.The results showed that B5 medium supplemented with 0.5mg/L NAA,0.6mg/L 6-BA and 30g/L sucrose,at initial pH 5.0～5.5,20g(FW)/L inoculation quantity and 110 r/min of rotation speed was a preferable culture conditions for E.ulmoides suspension cells growth and flavonoids synthesis.The results of metabolic kinetics analysis for E.ulmoides cell suspension culture showed that the logistic and Luedeking-Piret equations can be used for describing the kinetics of cell growth,sucrose consumption and flavonoids production during the process.The maximum specific growth rate(?m),the actual growth yield based on sucrose(YG) and maintenance coefficient(m) were 0.417/d,0.619g/g and 0.0206g/(g?d-1) respectively.All these outcomes could give a basis for establishing the suspension cell culture of E.ulmoides and production of the natural active components in large-scale.