Objective To explore the effects of H 2 S on neuronal injuries induced by oxygen glucose deprivation /reoxygenation ( OGD/R) in cortical neurons .Methods For OGD, the primary cultured cortical neurons were incubated with glucose-free EBSS media for 4h in N2/CO2/O2 (93%/5%/2%) atmosphere.Thereafter, the media were replaced by Neurobasal/B27 culture media and the neurons were incubated for 12 h in a 5%CO2 incubator at 37℃.NaHS was used as a H2S donor and cell survival rate was determined by cell counting kit 8(CCk-8).[Ca2+]i was determined using fura-2/AM and fluorescence microscopic imaging systems .The release rate of lactate dehydrogenase ( LDH) was determined by lactate dehydrogenase assay kit , and cell damage was analyzed by staining of propidium iodide ( PI ) .Results After pretreated with 200, 300 and 600μmol/L sodium hydrosulfide ( NaHS) for 30min before OGD/R, the cell survival rate of neurons significantly increased (n=4).[Ca2+]I(n=5), LDH release rate (n=4) and cell damage percentage (n=6) in the neuron pretreated with 300 μM NaHS were significantly lower than those in ODG/R cells.Treatment with 10μmol/L calcium chelator BAPTA also reduced the LDH release rate and cell damage percentage induced by ODG /R in neurons . Conclusion The results indicate that H 2 S may inhibit the OGD/R induced damage in cortical neurons via reducing calcium overload of neurons .

Objective To establish a rabbit model of restenosis and analyze the expressions of VEGFmRNA and TGF-?_1mRNA during the intimal proliferation.We also explored the relationship between VEGFmRNA,TGF-?_1mRNA and restenosis.Methods 40 healthy male New Zealand white rabbits were evenly divided into three injury groups and one control group.Right carotid arteries were injured with PCI balloon in the injury groups.10 rabbits of each injury group were sacrificed on weeks 1,2 and 4 after the injury.VEGFmRNA and TGF-?_1mRNA were examined by in situ hybridization.All the samples were analyzed using a computerized imaging analysis system.Results In the injury groups,neointimal areas were significantly larger than those in control group(P

Objective To explore the effect of micro-lectures withadvanced simulation man in improving the practical skills teaching of nursing students, so as to promote the students' post competency. Methods Totally the 186 nursing internswere divided into control group and observation group by random number method with 93 people in each group. The control group used the traditional teaching modein the teaching of practical skills.The observation group used the micro-lectures with high simulation teaching.Comparing the two groups of nursing students comprehensive assessment test simulation results in the theory, skills, scenarios, and nursing students the evaluation of the curriculum. Results The scores of the two groups were all above the qualification line, but the scores of the observation group were significantly higher than those of the control group (P<0.05). The theoretical examinations and the situation simulation comprehensive testswas(77.89 ± 7.79), (75.60 ± 7.92)points in control group, and (93.87 ± 3.90),(92.87 ± 4.08)points in observation group, there was significant difference between two groups (t=17.67, 18.70,all P=0.000). The curriculum evaluation results of improving learning initiative, active curriculum atmosphere, clear operation demonstration, exercise clinical thinking, improve the clinical interest were 81.72％(76/93), 72.04％(67/93), 93.55％(87/93), 60.22％(56/93), 67.74％(63/93)in control group, and 96.77％(90/93), 95.70％(89/93), 100.00％(93/93), 92.47％(86/93), 98.92％(92/93)in observation group, there was significant difference between two groups(χ2=20.39, P=0.016).Conclusions The effect is significant of micro-lectures combined with high simulation of human using in clinical skills teaching. Thismold can conducive to the cultivation of clinical thinking of nursing students, and improve clinical comprehensive ability, and promote the promotion of nursing students post competency.

Objective: To explore effects of dendritic epidermal T cells (DETCs) and Vγ4 T lymphocytes on proliferation and differentiation of mice epidermal cells and the effects in wound healing of mice. Methods: (1) Six C57BL/6 male mice aged 8 weeks were collected and divided into control group and wound group according to random number table (the same grouping method below), with 3 mice in each group. A 4 cm long straight excision with full-thickness skin defect was cut on back of each mouse in wound group, while mice in control group received no treatment. On post injury day (PID) 3, mice in 2 groups were sacrificed, and skin within 5 mm from the wound margin on back of mice in wound group and normal skin on corresponding part of mice in control group were collected to make single cell suspensions. The percentage of Vγ4 T lymphocyte expressing interleukin-17A (IL-17A) and percentage of DETCs expressing insulin-like growth factor Ⅰ (IGF-Ⅰ) were detected by flow cytometer. (2) Ten C57BL/6 male mice aged 8 weeks were collected and divided into control group and Vγ4 T lymphocyte depletion group with 5 mice in each group. Mice in Vγ4 T lymphocyte depletion group were injected with 200 g Vγ4 T lymphocyte monoclonal neutralizing antibody of Armenian hamster anti-mouse intraperitoneally, and mice in control group were injected with the same amount of Armenian hamster Ig intraperitoneally. One hole with full-thickness skin defect was made on each side of spine of back of each mice. The wound healing was observed on PID 1-8, and percentage of remaining wound area was calculated. (3) Six C57BL/6 male mice aged 8 weeks were grouped and treated in the same way as in experiment (2), with 3 mice in each group. On PID 3, expressions of IL-17A and IGF-Ⅰ in epidermis on margin of wound were detected with Western blotting. (4) Thirty C57BL/6 male mice aged 3 days were sacrificed, and epidermal cells were extracted. The keratin 14 positive cell rate was examined by flow cytometer (the same detecting method below). (5) Another batch of mouse epidermal cells were collected and divided into control group, IGF-Ⅰ group, and IL-17A group, with 3 wells in each group (the same well number below). Cells in IGF-Ⅰ group and IL-17A group were added with 1 mL recombinant mouse IGF-Ⅰ and IL-17A with final mass concentration of 100 ng/mL respectively, while cells in control group were added with the same amount of sterile phosphate buffered saline (PBS). On post culture day (PCD) 5, keratin 14 negative cell rate was examined. Another batch of mouse epidermal cells were collected, grouped, and treated in the same way as aforementioned experiment, and keratin 10 positive cell rate was examined on PCD 10. (6) Another batch of mouse epidermal cells were collected and added with 4 mmol/L 5(6)-carboxyfluorescein diacetate N-succinimidyl ester (CFSE) solution, and divided into control 0 d group, control 7 d group, IGF-Ⅰ group, and IL-17A group. Cells in IGF-Ⅰ group and IL-17A group were treated in the same way as the corresponding groups in experiment (5), and cells in control 0 d group and control 7 d group were treated in the same way as the control group in experiment (5). The CFSE fluorescence peaks were examined on PCD 0 of control 0 d group and PCD 7 of the other 3 groups. (7) Another batch of mouse epidermal cells were collected and divided into control group and IGF-Ⅰ group. Cells in IGF-Ⅰ group were added with 1 mL recombinant mouse IGF-Ⅰ with final mass concentration of 100 ng/mL, and cells in control group were added with the same amount of sterile PBS. On PCD 5, cells were underwent keratin 14 staining and CFSE staining as aforementioned, and keratin 14 negative cell rate of CFSE positive cells was examined. Another batch of mouse epidermal cells were collected and divided into control group and IL-17A group. Cells in IL-17A group were added with 1 mL recombinant mouse IL-17A with final mass concentration of 100 ng/mL, and cells in control group were added with the same amount of sterile PBS. On PCD 5, keratin 14 negative cell rate of CFSE positive cells was examined. Data were processed with one-way analysis of variance and t test. Results: (1) On PID 3, percentage of DETC expressing IGF-Ⅰ in normal epidermis of control group was (9.9±0.8)%, significantly lower than (19.0±0.6)% of epidermis around margin of wound group (t=8.70, P<0.01); percentage of Vγ4 T lymphocyte expressing IL-17A in normal epidermis of control group was (0.123±0.024)%, significantly lower than (8.967±0.406)% of epidermis around margin of wound group (t=21.77, P<0.01). (2) On PID 1-4, there was obvious inflammatory reaction around wounds of mice in control group, and on PID 5-8, the wound area was still large. On PID 1-4, there was slight inflammatory reaction around wounds of mice in Vγ4 T lymphocyte depletion group, and on PID 5-8, the wound area was significantly reduced. On PID 3-7, percentages of residual wound area in Vγ4 T lymphocyte depletion group were significantly lower than those in control group (t=5.92, 5.74, 7.17, 5.38, 5.57, P<0.01), while percentages of residual wound area in two groups on PID 1, 2, 6 were similar (t=1.46, 3.17, 3.10, P>0.05). (3) On PID 3, compared with those in control group, expression of IL-17A and IGF-Ⅰ in epidermis around wound margin of mice in Vγ4 T lymphocyte depletion group was markedly decreased and increased respectively (t=8.47, 19.24, P<0.01). (4) The keratin 14 positive cell rate of mouse epidermal cells was 94.7%. (5) On PCD 5, the keratin 14 negative cell rate of mice in control group was markedly higher than that of IGF-Ⅰ group, while significantly lower than that of IL-17A group (t=7.25, 5.64, P<0.01). On PCD 10, the keratin 10 positive cell rate of mice in control group was significantly higher than that of IGF-Ⅰ group, while significantly lower than that of IL-17A group (t=3.99, 10.82, P<0.05 or P<0.01). (6) Compared with that of control 0 d group, CFSE fluorescence peaks of mouse epidermal cells in control 7 d group, IGF-Ⅰ group, and IL-17A group on PCD 7 shifted to the left. Compared with that of control 7 d group, CFSE fluorescence peaks of mouse epidermal cells in IGF-Ⅰ group and IL-17A group on PCD 7 shifted to the left. (7) On PCD 5, keratin 14 negative cell rate of CFSE positive cells of mice in control group was significantly higher than that in IGF-Ⅰ group (t=9.91, P<0.01), and keratin 14 negative cell rate of CFSE positive cells of mice in control group was markedly lower than that in IL-17A group (t=6.49, P<0.01). Conclusions In the process of wound healing, IGF-Ⅰ secreted by DETC can promote the proliferation of mouse keratin 14 positive epidermal cells and inhibit their terminal differentiation, while IL-17A secreted by Vγ4 T lymphocyte can promote the proliferation and terminal differentiation of mouse keratin 14 positive epidermal cells, thus both IGF-Ⅰ and IL-17A can affect wound healing.