Objective:To study the expression of kringle 1-3 domain fragment of human plasmihogen in E.coli and the anti-tumor activity of its product.Methods:The K1-3 domain fragment was cloned in expression vector pBV220,the resulted recombinant plasmid pBV-K13 was transformed into E.coli DH5? and its product was purified and assayed its bioactivity.Results:K1-3 domain fragment was expressed in(E.coli) DH5?.The results showed the expressed product covered 20% of the total bacterial protein on SDS-PAGE and the Western blot analysis showed that the product had immunological specificity with the antiserum of human plasminogen and inhibits the growth of chorioallantoic membrane(CAM) angiogenesis and mouse B16 melanoma.Conclusion:Human plasminogen K1-3 domain fragment was expressed in E.coli;the expressed product has anti-angiogenesis and anti-tumor activity.

BACKGROUND:Cerebral hemorrhage can activate the proliferation and differentiation of neural stem cel s in the dentate gyrus of the hippocampus. Through continuous differentiation and proliferation, endogenous neural stem cel s can gradual y replace aging and damaged neurons, thus protecting the brain structure.
OBJECTIVE:To compare the difference of the proliferation and differentiation of neural stem cel s in the dentate gyrus of the hippocampus of rats with different ages.
METHODS:Ninety-six adult rats and 96 aged rats were randomly divided into normal group (n=18 per group), sham operation group (n=12 per group) and cerebral hemorrhage group (model group, n=66 per group), respectively. Cerebral hemorrhage models were made in the two model groups in which, the rats were subjected to cerebral hemorrhage for 6, 24, 48, 72 hours and 7 days, respectively. Then, brain tissues were col ected to measure brain water content. BrdU/NeuN and BrdU/GFAP double staining were performed at 3, 7, 14, 21, 28 days after surgery to calculate the number of positive cel s.
RESULTS AND CONCLUSION:For both adult and aged rats, the brain water content was significantly higher than that in the normal group and sham operation group (P<0.05), while in the normal and sham operation groups, the brain water content was significantly lower in the aged rats than the adult rats (P<0.05). The number of bilateral BrdU-positive cel s in the adult and aged model groups was significantly higher than that in the corresponding normal and sham operation groups (P<0.05), and moreover, the positive cel number at the hemorrhage side was significantly higher than that at the opposite side (P<0.05). In addition, the number of BrdU-positive cel s at the hemorrhage side in the adult rats was significantly higher than that in the aged rats at different time after cerebral hemorrhage (P<0.05). Results from immunohistochemical double staining showed that the BrdU/NeuN and BrdU/GFAP expression in the hippocampal dentate gyrus of adult rats with cerebral hemorrhage was significantly higher than that of normal adult rats. Al these experimental results show that there are a few neural stem cel s proliferating in the hippocampal dentate gyrus of normal rats, and the proliferation ability is stronger in the adult rats than the aged rats. Cerebral hemorrhage can significantly strengthen the proliferation of neural stem cel s in the dentate gyrus in the adult rats compared with the aged rats.

BACKGROUND:Previous studies showed that neurotrophic factor has a variety of functions, which can effectively maintain the survival of neurons after injury.
OBJECTIVE:To observe the effect of adenovirus-mediated brain-derived neurotrophic factor on the differentiation of endogenous neural stem cels after intracerebral hemorrhage in rats.
METHODS:A total of 90 Sprague-Dawley rat models of cerebral hemorrhage were made. At 12 hours after cerebral hemorrhage, 5-bromodeoxyuridine (BrdU) was intraperitonealy injected, twice a day, for 10 consecutive days. After model establishment, rats were randomly divided into three groups, 30 rats in each group, and were respectively subjected to brain stereotaxic injection of adenovirus vector, adenovirus-mediated brain-derived neurotrophic factor and physiological saline. At 1 day, 3 days, 1 week, 2 weeks, 3 weeks, and 4 weeks, neurological deficit score was evaluated. Absorbancevalue of growth associated protein around the area of hematoma after intracerebral hemorrhage was measured. At 4 weeks after injection, double immunostaining was used to detect the expression of BrdU/NeuN and BrdU/glial fibrilary acidic protein (GFAP).
RESULTS AND CONCLUSION:(1) With the passage of time, nerve function defect score decreased in the three groups. At 1-4 weeks after injection, nerve function deficit scoreswere lower in the adenovirus-mediated brain-derived neurotrophic factor group thanthat in the adenovirus vector group and saline group (P< 0.05). (2) With the passage of time, the average absorbance of three groups in the peri-hematoma region first increased and then decreased. The absorbance value was higher in the adenovirus-mediated brain-derived neurotrophic factor group than in the adenovirus vector group and saline group at 3 days-4 weeks (P< 0.05). (3) BrdU/NeuN and BrdU/GFAP rates were significantly higher in the adenovirus-mediated brain-derived neurotrophic factor group thanthat of adenovirus vector group and saline group (P< 0.05). (4) The results show that the brain-derived neurotrophic factor mediated by adenovirus, and intervention on cerebral hemorrhage in rats can effectively promote the differentiation of endogenousneural stem cels, and promote the recovery of neural function in animal.

Objective To observe the effects of Tanshinone Ⅱ A sodium sulfonate (TSS ) on ischemia/reperfusion (I /R) induced cardiac injury in male (Sprague-Dawley,SD ) and explore its mechanisms.Methods Rats were subjected to a 30 min coronary arterial occlusion followed by 24 hours reperfusion.The survival rats were randomly (random number)divided into sham group (Sham group,n =10),ischemia reperfusion group (I /R group,n =10),low dose of TSS group (TSS-L group,n =10), medium dose of TSS group (TSS-Mgroup,n =9),high dose of TSS group (TSS-H group,n =9).A MAP heart function analysis system was used to measure hemodynamic variables,and TTC staining method was used to detect the myocardial infarct size.The levels of Bcl-2,Bax,Caspase-3,Lc3B/Lc3A,Beclin-1 and high mobility group box1 (HMGB1)were detected by western blot method.All data were analyzed by using One-way analysis of variance (ANOVA)(LSD-t test).Results Cardiac function in I /R group was lower than that in Sham group,and that was significantly improved by pretreated with TSS (P <0.05),but there were no significant differences in Pmin and DAP between Sham group and I /R group (P >0.05 ).The percentage of myocardial infarct size in TSS pretreatment group was significantly smaller than that in I /R group (P <0.05 ).Compared with Sham group,levels of Caspase-3 and Bax increased,and the Bcl-2 content was reduced obviously in I /R group (P <0.05).TSS pretreatment significantly down-regulated the levels of Caspase-3 and Bax protein (P <0.01).At the same time,the level of Bcl-2 was increased in all TSS pretreatment groups (P <0.01).Compared with Sham group,the ratio of Bcl-2 /Bax in I /R group was lower (P <0.05),and that was elevated in TSS groups (P <0.05 ).The change of autophagy related protein beclin-1 and Lc3B/Lc3A was in similar trend,and the levels of beclin-1 and Lc3B/Lc3A in I /R group were lower than that in Sham group (P <0.05),and those were raised in TSS pretreatment groups (P<0.05).The level of HMGB1 in I /R group was higher than that in Sham group (P <0.05),and compared with I /R group,the level of HMGB1 significantly decreased in TSS pretreatment groups (P <0.01 ). Conclusions The tanshinone ⅡA sodium sulfonate can protect the myocardium from ischemia/reperfusion injury and the mechanism may be attributed to the inhibition of cell apoptosis and activation of cell autophagy.

Aim Tostudytheanalgesiceffectofoxyso-phoridine (OSR)on GABA transporter-1 (GAT-1 )mR-NA expression and its influence on GAT-1 expression inmice.Methods Formalintestwasusedtodetectthe analgesic effect of OSR(iv).Immunohistochemis-try was taken to inspect the expression of GAT-1 in cerebral cortex and thalamus in mouse brain. The quantitative real-time PCR method was used to inspect the influence of OSR on GAT-1 mRNA expression of braininmice.Results OSR(500,250,125mg· kg-1 ,iv ) could significantly increase the foot-licking latency.OSR(500 mg·kg-1,ip)could significantly decrease the number of GAT-1 immuopositive cells incerebral cortex and thalamus in mouse brain,and re-duce GAT-1 mRNA expression in brain(P<0. 01,P<0.05)intheformalintest.Conclusion OSRhasa significant analgesic effect,and its analgesic mecha-nism is related to the GAT-1 expression in mouse brain.

AIM: Constructing plasmid that expresses human plasminogen kringle 5 gene to analyze the gene expression in E.coli. METHODS: The gene of human plasminogen kringle 5 was inserted into plasmid PBV220 EcoRI site by gene manipulation techniques and was transformed to E.coli TGI. The gene expression was observed by SDS-PAGE. RESULTS: Expression vector PBVK5 was constructed, and human plasmingen kringle 5 gene product was obtained at 42℃ induction. CONCLUSLON: Expression product of human plasminogen kringle 5 gene was soluble form of proteins, and the expression amount was 9.8% in E.coli TGI total proteins.

ObjectiveTo explore the effect of Funaoling Oral Liquid on improving memory function of mice.Methods30 mice were injected with scopolamine 1mg/kg to make model of memory obstacle after treated by Funaoling Oral Liquid,piracetam or saline for 10 days.The escape test was used to measure the memory function.The blood circulation obstacle of the mice in mesentery was made by norepinephrine,and then, Funaoling Oral Liquid, Happy-hundred-year Oral Liquid and saline were given to observe the recovery time. The serum superoxide dismutase(SOD) activity in mice was measured after using different dosage of Funaoling Oral Liquid(75g/kg,37.5 g/kg and 18.75g/kg),piracetam or saline for 10 days.ResultsFunaoling Oral Liquid improved the memory function and blood circulation of mesentery significantly compared with that of the model(P<0.05 or P<0.01). The serum SOD activity in mice which was treated by Funaoling Oral Liquid 18.75g/kg were higher than that of model(P<0.05).ConclusionsThe Funaoling Oral Liquid can improve the memory and blood circulation of mice after injured,as well as to enhance the activity of SOD.

Objective To observe the effect of interactive somatic game Kinect on motor of lower limbs and balance function in patients with stroke. Methods 40 inpatients were divided into experimental group (n=20) and control group (n=20). All the patients received routine rehabilitations, while the experimental group received the interactive game Kinect in addition. They were assessed with Fugl-Meyer assessment of lower limb (FMA) and Berg balance scale (BBS) before and after intervention. Results The scores of flexor synergy, extensor synergy,movement combining synergies (in sitting), lower limb total score of FMA, and the score of BBS increased more in the experimental group than in the control group (P<0.05) after intervention. Conclusion The interactive somatic game Kinect can further improve the motor of lower limb and balance function of patients with stroke.

Objective To investigate the effects of dexmedetomidine (DEX) on the mRNA expression of triggering receptor expressed on myeloid cells 1 (TREM-1) in peripheral blood mononuclear cells of rats with acute obstructive suppurative cholangitis (AOSC).Methods Sixty healthy male Wistar rat models of AOSC induced by complete common bile duct ligation and injection of E.coli into the bile duct through an intubation tube were replicated successfully.After modeling,the peripheral blood was collected and mononuclear cells were isolated and cultured.According to random number table method,the mononeuclear cells were divided into model group (no drug added in culture of mononuclear cells) and low,medium and high dose DEX groups (final concentrations 0.4,0.8,1.2 μg/L DEX were in low,medium and high DEX mononuclear cell cultures,respectively).After the mononuclear cells were cultured for 24 hours,the levels of tumor necrosis factor-α (TNF-α),interleukins (IL-1 and IL-6) in the supernatant of the cultured mononuclear cells were detected by enzyme linked immunosorbent assay (ELISA).The level of C-reactive protein (CRP) was detected by immunity transmission turbidimetry.The expression of TREM-1 mRNA in the mononuclear cells was detected by reverse trantscription-polymerase chain reaction (RT-PCR).Results Compared with the model group,the levels of TNF-α,IL-1,IL-6,CRP were decreased,the TREM-1 mRNA expressions were down-regulated in the different DEX dose groups,and the degrees of descent in medium and high dose groups were more significant than those in low dose group [TNF-oα (ng/L):95.5±8.6,88.9±5.3 vs.131.1 ± 14.2;IL-1 (ng/L):53.5±8.3,48.3 ± 6.7 vs.73.7 ± 12.8;IL-6 (ng/L):266.9±26.2,252.1 ± 17.7 vs.349.9±40.4;CRP (ng/L):4.3 ± 1.1,3.9 ±0.7 vs.5.6 ± 1.7;TREM-1 mRNA (A value):0.43 ± 0.18,0.39 ± 0.16 vs.0.65 ±0.25,all P ＜ 0.05].Conclusion DEX can down-regulate the expression of TREM-1 mRNA and inhibit the formation and secretion of inflammatory factors TNF-α,IL-1,IL-6 and CRP in peripheral blood mononuclear cells of rats with ASOC.