Objective: To investigate the starvation-induced autophagy in rat C6 glioma cells and the changes of autophagy-related protein Beclin-1. Methods: Rat C6 glioma cells of the logarithmic growth phase were cultured with amino acid-free EBSS medium instead of the DMEM medium. Cells were collected at 1 h, 3 h, 6 h, 12 h, and 18 h after EBSS culture. Western blotting analysis and immunofluorescence were used to detect the specific markers of autophagic microtubule-associated protein 1 light chain 3 (LC-3) and autophagy-related protein Beclin-1, and transmission electron microscope was used to observe the autophagic body. Results: LC-3 expression was found in C6 glioma cells 1 h after starvation; the expression increased with the prolongation of starvation, and reached a peak 12 h later and then began to decline. Autophagic bodies were observed under transmission electron microscopy, and autophagosomes were found in immunofluorescence. Beclin-1 expression began to increase in C6 glioma cells 1 h after starvation, reached a peak at 3 h and then decreased gradually. Increased expression of Beclin-1 was also observed by immunofluorescence. Conclusion Starvation can induce autophagy in C6 glioma cells, which is correlated with the expression of autophagy-related protein Beclin-1. This study lays a foundation for studying the autophagy mechanism in glioma cells.

Biocompatible Materials ; analysis ; Chromatography ; methods ; Chromatography, Gas ; methods ; Chromatography, Liquid ; methods ; Paraquat ; analysis ; Pesticide Residues ; analysis

Adult ; Heart Aneurysm ; congenital ; Heart Atria ; pathology ; Humans ; Male

Adolescent ; Cerebellar Diseases ; diagnosis ; drug therapy ; etiology ; Cerebellum ; physiopathology ; Child ; Child, Preschool ; Cognition Disorders ; diagnosis ; drug therapy ; etiology ; Female ; Humans ; Infant ; Male ; Mood Disorders ; diagnosis ; drug therapy ; etiology

Objective: To explicit whether miR-200b can suppress the proliferation and invasion by targeting CD133, and to explore a molecular mechanism that miR-200b plays a role in tumor suppression in glioma. Methods: The CD133 3'-untranslated region (3'-UTR) mRNA-luciferase reporter vector was constructed and the dual-luciferase reporter gene assay was employed to examine the effect of miR-200b on activity of luciferase. U251 cells were transfected with miR-200b mimics and CD133-small interfering RNA (siRNA) (as a positive control) by LipofectAMINE 2000, and the expressions level of CD133 protein was detected by Western blotting. The inhibition effects of CD133 on cell proliferation and invasion were observed after CD133 siRNA were transfected into U251 cells. U251-SC cell mammosphere assay was performed after cotransfection with miR-200b mimics and CD133 siRNA. Results: miR-200b could bind to the 3'-UTR of CD133 and inhibit the activity of luciferase. CD133 protein expression was significantly down-regulated when miR-200b was overexpressed in U251 cells. Overexpression of miR-200b inhibited the invasion of U251 cells. Overexpression of miR-200b antagonized a role of proliferation and invasion in U251 cells. Overexpression of miR-200b reduced mammosphere number and the size of glioma stem cells. Conclusion: miR-200b can suppress cell proliferation and invasion by targeting CD133 in glioma. Copyright © 2014 by TUMOR.

AIM:To investigate the value of pattern visual evoked potential (PVEP) and flash electroretinogram (FERG) in early diagnosis and prevention of diabetic retinopathy (DR), analyzing the correlation of early stage DR with PVEP and FERG.
METHODS: Sixty patients, 30 males and 30 females, participated in observation group. Their average age was 19. 42 ± 7. 78years. The duration of DM was < 5a. Best corrected visual acuity was 5. 0. Fasting blood glucose was 7. 8± 3. 6mmol/ L. There were 60 subjects, 30 males and 30 females, in control group. Their average age was 17. 2 ± 6. 52years. Best corrected visual acuity was 5. 0. Every participator was tested with PVEP and FERG according to ISCVE standard. The amplitude of PVEP and P100 latency were recorded. And the b-wave latency, b-wave amplitude, a - wave latency, a - wave amplitude were showed down.
RESULTS: In observation group, P100 amplitude decreased and P100 latency increased, compared to those of control group ( P< 0. 01); b - wave latency, b -wave amplitude, a - wave latency, a - wave amplitude were different from those in control group(P<0. 01); the fasting blood glucose kept stable; P100 amplitude, b -wave amplitude and a-wave amplitude were not related to the DM duration; P100 latency, a-wave latency and b-wave latency were related to the DM duration.
CONCLUSION: PVEP are sensitive to optic neuron damage; FERG is desirable to detect the lesion of Müller cells and bipolar cells. P100 amplitude by PVEP, b-wave amplitude by FERG may be the most sensitive parameter for DR at early stage.

Bacillus subtilis ZC-7 was obtained by implantation with N+ ions beam to B.subtilis AS1.398,and compared with the AS1.398 neutral protease,the enzyme activity of ZC-7 neutral protease was about 1 timeshigher in previous research.A neutral protease was purified from the culture of B.Subtilis ZC-7 by the procedures including amoninium sulfate precipitation,ultrafiltration,DEAE-Sepharose Fast Flow chromatography and Sephadex G-75 chromatography.By multi-step purification,the ZC-7 neutral protease was purified to 78.5 folds and its yield was 27.7%,at last,the specific activity of ZC-7 neutral protease was up to 4.1?105U/mg.Analysed by SDS-PAGE,the purified protease has shown a molecular mass of about 42kDa.The Km for casein hydrolysis was 3.67?10-3?g/ml and the Vmax was 12.21?g/min.The optimum pH and temperature forhydrolysis of casein were 7.0 and 55℃,respectively.This protease was stable up to 40℃ within the pH range of 6.5 and 8.0.EDTA,isopropanol and alcohol nearly inhibited its activity while some ions such as Ca2+,Mg2+,Fe3+ can improve its activity.In addition,it could resist 1 mol/L H2O2.

This study is to investigate the effect of 8-prenylnaringenin (8-PNG) on osteoclastogensis of bone marrow cells and bone resorption activity of osteoclasts. Osteoclasts were separated from long bone marrow of newborn rabbits and cultured in alpha-MEM containing 10% FBS. 8-PNG was added into culture media at 1 x 10(-7), 1 x 10(-6), 1 x 10(-5) mol xL(-1), separately. 17beta-Estradiol (E2, 1 x 10(-7) mol x L(-7)) was used as positive control. T RAP staining and TRAP activity measurement were performed after 5 days, and the bone resorption pits were analyzed after 7 days. Annexin V staining for the detection of apoptotic osteoclasts was performed after 2, 4, 8, 12, 24, 36 and 48 h separately. The mRNA expression level of TRAP and cathepsin K (CTSK) was measured by real-time RT-PCR. 8-PNG significantly reduced the number of osteoclasts which was TRAP staining positive and with more than three nucleus, the area and number of bone resorption pits decreased obviously in 8-PNG-supplemented groups. The apoptosis rate peaked earlier in the 8-PNG-supplemented groups and the mRNA expression level of TRAP and CTSK decreased significantly. All these inhibitory effects were in a dose dependent manner, the highest effect was obtained by 1 x 10(-5) mol x L(-1) 8-PNG. 8-PNG inhibits bone resorption activity of osteoclasts by inducing osteoclast apoptosis and inhibiting the gene expression and enzyme activity including TRAP and CTSK, and restrains bone marrow cells to osteoclast differentiation.

OBJECTIVETo compare mid-term clinical outcomes between amputation and limb salvage in treating severe open tibial fractures with type Gustilo III B, III C.

METHODSFrom July 2007 to June 2010,68 patients with severe open tibial fractures with type Gustilo III B, III C treated by amputation and limb salvage were retrospectively analyzed. In amputation group, there were 26 males and 12 females with an average age of (44.9±16.3) years old; and 21 cases were type Gustilo (III B, 17 cases were Gustilo III C; amputation were performed in accordance with soft tissue injury degree of shank, fracture types and surgical exploration. In limb salvageg group, there were 21 males and 9 females with an average age of (43.5±14.7) years old; and 23 cases were type Gustilo III B, 7 cases were Gustilo III C; the method of internal fixation and and wound healing were performed in accordance with patients's specific condition. Operative time, blood loss, hospital stay and postoperative infection was compared between two groups; time of loading and rate of return to work was compared; VAS scoring was used to evaluate condition of pain; SF-36 health queationaire was used to assess postoperative life quality.

RESULTSTotally 60 patients were followed up (33 cases in amputation group and 27 cases in limb salvage group) with an average time of 49.1 months. Operative time, blood loss, hospital stay and postoperative infection in amputation and limb salvage group respectively was (109.0±25.7) min, (245.0±58.6) min; (168.0±49.0) ml, (311.0±137.0) ml; (13.8±2.7) d, (28.8±13.1) d; 7.9%, 36.7%. At the final following-up, there was no significance meaning between two groups in VAS scoring and rate of return to work, but time of loading in amputation group was shorter than that of in limb salvage group. Physiological function in amputation group was better than limb salvage group, while body pain was worse; and there was no signicance meaning in psychological health between two groups.

CONCLUSIONAmputation and limb salvage both can treat severe open tibial fractures, and mid-term clinical outcomes between two groups has equivalent efficacy.

Adolescent ; Adult ; Aged ; Amputation ; methods ; Case-Control Studies ; Female ; Humans ; Limb Salvage ; methods ; Male ; Middle Aged ; Tibial Fractures ; surgery

OBJECTIVETo study immunomodulating activity of Lonicera Japonica flavone by investigating immune enzymatic activity of serum and antoxidized activity of lymphoid organs in mice.

METHODSFifty KM mice were randomly divided into control group, model group, low dose group, middle dose group and high dose group(n = 10), respectively. And low dose group, middle dose group and high dose group were given Lonicera Japonica flavone with 100 mg/kg, 200 mg/kg and 400 mg/kg every day, respectively, while control group and model group were administered with NS. After continuously giving drug 7 weeks, other groups were injected with Dexamethasome (Dex: 25 mg /kg) for 3 days by subcutaneous injection, but the control group were treated with NS. And after giving Lonicera Japonica flavone 1 week simultaneously, organ indexes , the activity of acid phosphatase (ACP), alkaline phosphatase (AKP) and lysozyme (LSZ) in serum , and the content of monoamine oxidase (MAO), total antioxidant capacity (T-AOC), total superoxide dismutase (SOD) and malondialdehyde (MDA) in lymphoid organs in mice were tested, respectively.

RESULTSLonicera Japonica flavone could significantly improve the organ indexes, and significantly improve the activity of ACP, AKP and LSZ in serum, and significantly improve the contents of T-AOC and SOD, but reduce that of MAO and MDA in lymphoid organs in immunosuppressed mice.

CONCLUSIONIonicera Japonica flavone can significantly improve the activity of immune enzyme in serum and the antioxidized activity of lymphoid organs in mice. It suggests that Ionicera Japonica flavone has a good immunomodulatory effects.

Acid Phosphatase ; blood ; Alkaline Phosphatase ; blood ; Animals ; Antioxidants ; metabolism ; Flavones ; pharmacology ; Immunomodulation ; Lonicera ; chemistry ; Malondialdehyde ; metabolism ; Mice ; Monoamine Oxidase ; metabolism ; Muramidase ; blood ; Superoxide Dismutase ; metabolism