Objective To explore the effects of constraint-induced movement therapy (CIMT) on the expression of tyrosine hydroxylase (TH) and glial cell derived neurotrophic factor (GDNF) in Parkinson's disease (PD) model rats. MethodsPD models were established by microinjection of 6-hydroxydopamine (6-OHDA) solution into substantia nigra of rats' right cerebral hemisphere.Forty-two model rats were divided randomly into an exercise group and a control group 1 week after microinjection.The exercise group rats were forced to use their impaired limbs by placing their nonimpaired fore-limbs in casts.The control group rats were housed in the same environment without any special treatment.Two weeks after 6-OHDA infusion and exercise training,the behavioral changes of rats were examined after intraperitoneal injection apomorphine ( APO).The content of dopamine (DA) and dihydroxyphenylacetic acid (DOPAC) was measured by high performance liquid chromatography with electrochemistry ( HPLAEC) ; the expressions of TH and GDNF in striatum were detected by immunohistochemical methods and TH,GDNF mRNA were detected by reverse transcription-polymerase chain reaction (RT-PCR).Results After 2 weeks of training,the rotating laps of the rats in exercise group within 30 min after APO induction,reduced to a significantly greater extent when compared to the control group (P ＜ 0.05).The content of DA and it's metabolites DOPAC in striatum homogenate was significantly higher in exercise group than that in the control group ( P ＜ 0.05 ),and the expression levels,of TH and GDNF protein/ mRNA were also significantly higher in the exercise group than those in control group ( P ＜ 0.05 ).Conclusions CIMT can improve the behavioral performance of PD rats,probably through promoting the expressions of TH and GDNF protein/mRNA in striatum,and increasing DA and it's metabolites DOPAC level.

Objective To investigate the clinical effects of two minimally invasive surgical treatments for billary acute pancretitis.Method In this study,63 patients with billary acute pancretitis were prospectively divided into two groups.Patients in group A received laparoscopic cholecystectomy (LC) and laparoscopic transcyctic common bile duct exploration (LTCBDE) within 72 hours of onset,group B underwent endoscopic nasobiliary drainage (ENBD).The two groups were evaluated by blood amylase and urine amylase,alanine aminotransferase,aspartate aminotransferase on postoperative day 1,3,5,local complication and the recurrence rate within 6 months.Result Blood amylase was lower in group A than that in group B (P =0.04) on postoperative day 1.There were no significant differences in amylase,aminotransferase of two groups on postoperative day 3,5 (all P ＞ 0.05).The incidences of local complications and pancreatic necrosis infection were 2.7％,7.7％ respectively(P =0.13) ; the incidences of pancreatic pseudocyst were 5.4％,7.7％ (P =0.42) ; the recurrence rate within 6 months were 2.7％ and 19.2％ respectively(P =0.006).Conclusions The clinical curative effects of early LC and LTCBDE or ENBD on billary acute pancretitis showed no significant differences,but the recurrence rate of acute pancretitis within 6 months revealed that early LC and LTCBDE may be more effective.

Objective To build of lentiviral mediated RNA interference of mammalian target of rapamcin (mTOR) ,because the mammalian target of rapamycin plays a important role in tumor development and the signal path .Methods According to RNA in-terference (RNAi) design rules Completely ,in view of the gene called mTOR was desiged four interference targets and Negative control (FAM ) sequence ,first of all ,synthetic oligonucleotides nucleotide fragments with artificial ,and can obtain siRNA fragments effectively by the method of PCR joining together ,then undertake transfection on lung adenocarcinoma A 549 cell by Lipofectamine transfection reagent 2000 .To begin to observe the enhanced green fluorescent protein expression in lung adenocarcinoma A 549 cells by fluorescence microscopy at high magnification after 1 days .We can use semi-quantitative RT-PCR method ,and detect of mTOR gene expression of mRNA level after 1 days ,meanwhile ,testing the expression of protein levels by Western Blot after 2 days ,in or-der to select the most efficient interference target sequence ,,afterwards ,synthetic double-stranded DNA ,and it can be make up vec-tor system with plasmid pHelper 1 .0 and pHelper 2 .0 by the pGCL-GFP carrier ,further transfect 293 T cells ,at last produce lenti-viral after packaging ,then detection GFP protein expression levels by Western-Blot method ,and consequently detect the virus drops degree of 293T cells ,at the same time ,identify the activity .Results The high efficiently target of mTOR gene has been successfully selected ;mTOR siRNA infecte of human lung adenocarcinoma A549 cells in vitro culture ,and the gene is obviously silence no mat-ter from the mRNA level or protein level ;3 mTOR gene lentivirus siRNA carrier was successfully build .geting virus supernatant al-so ,and virus drops to 1 × 108 UT/mL .Conclusion MTOR siRNA infected of human lung adenocarcinoma A549 cells in vitro cul-ture ,and could lead to mTOR gene obviously silence ;The construction was successfully gene mTOR siRNA lentivirus vectors .

Objective To reveal the effect of lutein on the status of oxidative stress in rats with high homocysteine levels (HHcy) and relevent molecular mechanisms.Methods The wistar rat HHcy model was established by intra-gastric administration with L-methionine suspension and treated with lutein.The oxidative stress status and the gene expression changes of transcription Nf-E2-related factor2 (Nrf2),a regulation factor of down stream antioxidant protein gene expression,were detected in HHcy rats and lutein intervention rats.Results Compared with the rat serum SOD activities (134.32 ± 12.65) U/mL in the control group,the serum SOD activities in the HHcy model group (95.6 ± 10.92) U/mL were significantly lower (P ＜ 0.05).The serum glutathione peroxidase (GPx) activities in the HHcy model group (121.66 ± 18.64) U/mL were also significantly lower as compared with the the control group (183.17 ± 21.29) U/mL,P ＜ 0.05.However,the serum SOD (126.75 ± 11.26) U/mL and GPx activities (167.18 ± 19.66) U/mL in the lutein intervention group were significantly higher as compared with the HHcy model group.As compared with the rat serum malondialdehyde (MDA) content (5.11 ± 0.68) μmol/L as well as the hydroxyl free radical levels (0.53 ± 0.05) U/L in the control group,the serum MDA content (7.65 ± 0.87) μmol/L and the hydroxyl free radical levels (0.92 ± 0.09) U/L in the HHcy model group were significantly higher.The serum MDA content (6.44 ±0.91) μmol/L and the hydroxyl free radical levels (0.74 ± 0.06) U/L in the lutein intervention group were significantly lower as compared with the HHcy model group.RT-PCR and Western blot results also showed decreased expression of SOD2 and GPxl mRNA in aorta epithelial tissues of HHcy model rats.With lutein intervention,the expressions ofSOD2 and GPx1mRNA were significantly increased and the gene expression of Nrf2 also up-regulated.Conclusions The Hhcy model rats were under the status of augmented oxidative stress,and carotenoid lutein could attenuate the Hcy-mediated oxidative stress,and its mechanism might be potentially associated with up-regulating the expression of Nrf2,thereby inducing the expression of its downstream antioxidant proteins.

OBJECTIVE:To discuss the significance of passing GSP authentication of mono-drugstores by joining in the drugstore chain.METHODS:The status quo of GSP authentication was surveyed so as to find the merits of the GSP authenti-cation-passed drug chain retailers and the drug chain wholesalers.RESULTS&CONCLUSION:The mode of achieving the passage of GSP authentication of the mono-drugstore by joining in the drug store chain is feasible and essential.

Objective To evaluate the value of covered self-expending metallic stent in the management of malignant esophageal and duodenal strictures or fistulas and explore the treatment for the associated common complications.Methods With interventional procedure under fluoroscopic guidance,95 self-expending mesh stents were implanted in 87 patients,including esophageal carcinoma(50 cases),esophagogastric anastomotic stenosis(23 cases),gastroduodenal malignant stenosis(5 cases),esophagobronchial fistula(3 cases),esophagogastric anastomotic fistula(6 cases).Results Stent placement was successful in all patients at the first trial,having no instant complication except one acute bleeding during the procedure and all of them with relief of stenostic complaint,restoration of clictelic function and closure of fistulas.The long-term curative effect showed something to do with different kind of disease,and the related treatment.Conclusion Self-expending metallic covered mesh stent is safe,effective and less complication;and could be further promoted with addition of radiotherapy and(or)chemotherapy.

Objective To evaluate the value of covered metallic stent and arterial embolization in the management of pseudoaneurysm of extremities or kidneys.Methods Endovascular covered stent graft was inserted for isolation of peripheral extremity pseudoaneurysm in 3 patients and intraarterial embolization was performed with gelfoam in 2 patients for pseudoaneurysm in kidney.Results All 5 pseudoaneurysms kept occluded after the treatment.Distal arterial pulse was normal in 2 patients treated with stent graft;new thrombosis formed within the stent during the procedure but disappeared after thrombolytic therapy through catheter in 1 case;and still another new pseudoaneurysm presented at puncture site in 1 other case due to other factors.Conclusions Pseudoaneurysms in extremities or kidneys can be effectively treated with interventional techniques of covered stent graft and percutaneous arterial embolization.The long-term investigation is needed.

Objective To establish an orthotopic osteosarcoma nude mice model that co-expresses green fluorescent protein( GFP) , red fluorescent protein ( RFP) and luciferase for the purpose of monitoring the growth of osteosarcoma and screening drug candidates against osteosarcoma .Methods Human osteosarcoma cells of U 2-OS were infected with lentivirus carrying reporter gene .The reporter gene expression was verified by fluorescent microscopy and bioluminescence imaging.The cells were transplanted into tibia of the nude mice and monitored by bioluminescence imaging .Results The reporter gene was stably expressed in U 2-OS cells.The growth and metastsis of osteosarcoma could be detected in nude mice.Conclusion The established orthotopic osteosarcoma nude mice model is an ideal model for investigating the mechanism of growth and metastasis of osteosarcoma and for screening drug candidates against osteosarcoma .

Objective Non-muscle myosin heavy chain 9 (MYH9) plays an important regulatory role in the development of tumor.This study aimed to explore the expression of MYH9 in osteosarcoma tissues and its effects on epithelial-mesenchymal transition and invasion of osteosarcoma cells.Methods We collected 52 cases of osteosarcoma tissues and para-carcinoma tissues at 5 cm form the edge of the tumor.RT-PCR and immunohistochemistry were used to analyze the expression level of MYH9 mRNA and protein in the osteosarcoma tissues and para-carcinoma tissues.MYH9 shRNA plasmid was transfected into U2-OS cells to silenced the expression of MYH9, after transfected, the cells were divided into three groups: the normal U2-OS cells were the control group, the U2-OS cells transfected with empty plasmid were the empty group and U2-OS cells transfected with MYH9 shRNA were interference group.RT-PCR was used to detect the changes of MYH9 mRNA levels in the U2-OS cells, the protein level of MYH9, EMT related protein E-cadherin and Vimentin were detected by Western blot, and the ability of cell invasion was evaluated by Transwell assay.Results The results of RT-PCR showed that the relative expression MYH9 mRNA in para-carcinoma tissues(1.526±0.148) was significantly lower than that in cancer tissues (3.547±0.195) (P<0.05).The results of immunohistochemistry showed that MYH9 protein was mainly expressed in cytoplasm, and the expression in cancer tissues was significantly higher than that in para-carcinoma tissues, the positive expression rate were 59.6%(31/52) and 26.9%(14/52) respectively, the difference was statistically significant(P<0.05).The results of Western blot showed that the relative expression of MYH9 mRNA in interference group was significantly lower than that in control group and empty group (P<0.05) after silenced MYH9 gene, and compared with the control group, the E-cadherin in U2-OS cells was significantly up-regulated but the Vimentin was down-regulated.After 48h, all of the groups had cells through the microfiltration membrane, the numbers of cells through the microfiltration membrane in interference group(41.2±15.1) was significantly lower than that in control group(117.3±12.4) and empty group(193.5±14.7) (P<0.05).Conclusion The expression of MYH9 protein in osteosarcoma tissues was significantly higher than that in para-carcinoma t tissues, silenced MYH9 gene can reduce the invasive ability of osteosarcoma by reducing the epithelial interstitial transition.

Background Retinal neovascular diseases affect visual function.Although many drugs have been used to manage the visual diseases,their effectiveness is less than satisfactory.Studies showed that ursolic acid has multiple biological effects including anti-vascularization.However,the effect of ursolic acid on retinal neovascular diseases is unclear now.Objective This study was to observe the inhibitory effect of ursolic acid on the high oxygen-induced mouse retinal neovascularization after intravitreal injection.Methods Sixty clean 7-day-old C57BL/6J mice were divided into the blank control group,PBS control group,positive control group (triamcinolone) and low,moderate and high dose (1.5,3.0 and 6.0 μg) ursolic acid groups randomly.The blank control group mice were raised in normal environment,and the mice from other groups were fed in the environment with O2 concentration at (75±2)％ for 5 days together with the maternal mice.The mice then were back to the normal air environment to induce retinal neovascularization.Then,the drugs were intravitreally immediately injected in the mice of the different groups.The mice were sacrificed at the 17-day old for the preparation of retinal sections.Retinal new blood vessel was examined by haematoxylin and eosin stain under the light microscope,and the number of vascular endothelial cell nucleus breaking the inner limiting membrane was counted.The gene expressions of vascular endothelial growth factor (VEGF),cyclooxygenase-2 (COX-2) and matrix metalloproteinase-2 (MMP-2) in the mouse retinas were quantitatively assayed using reverse transcription PCR.Results The number of endothelial nuclei newly-generated vessel breaking internal limiting membrane in the mice of PBS control group was (18.65±3.24)/field,which was more than (0.78±0.11)/field of the blank control group obviously (t =2.24,P＜0.05).The number of endothelial nuclei newly-generated breaking internal limiting membrane in the moderate-or high-dose ursolic acid group was less than that of moderate group obviously,it was statistically significant(P＜0.05).The number of vascular endothelial cell nuclei breaking internal limiting membrane in high high-dose group was (13.32 ± 1.87)/field and (8.93 ± 1.09) /field,showing significant decreases in comparison with the PBS control group and low-dose ursolic acid group (18.65±3.24)/field (15.44±2.02)/field (all at P＜0.05).However,no significant difference were seen in the number of new vascular endothelial cell nucleus between the high-dose ursolic acid group and the positive control group(9.14±1.13)/field (t=1.17,P＞0.05).The relative expressions of COX-2 mRNA,VEGF mRNA and MMP-2 mRNA in the mouse retinas were higher in the PBS control group than those in the blank control group (t =13.45,12.49,14.32,all at P＜0.05),and those in the moderate-dose or high-dose ursolic acid group were lowed in comparison with the PBS control group and the low-dose ursolic acid group (all at P＜0.05),but there were no significant differences between the high-dose ursolic acid group and the positive control group (all at P＞0.05).Conclusions Ursolic acid can suppress retinal neovascularization by down-regulating the expressions of VEGF,COX-2 and MMP-2 in oxygen-induced retinopathy of mouse in dose-dependent manner.