OBJECTIVE: To clone, express and purify a fusion protein IVT, and detect its biological activity. METHODS: A fusion gene IVT with immunal function, anti-angiogenesis and apoptosis-inducing activities was constructed by genetic engineering means. pPIC9-IVT was constructed and transformed into Pichia pastoris GS115(his4). The recombinant fusion protein IVT was expressed in yeast engineering strain and purified from the culture supernatant. MTT assay was used to observe the effect of the IVT on the proliferation of CTLL-2 cells, human umbilical vein endothelial cells(HUVECs)and NCI-H446 cells. Transwell migration assay was used to observe the impact of the IVT on cell migration ability of HUVECs. The apoptosis of NCI-H446 cells was examined by Hoechst staining. RESULTS: The IVT promoted CTLL-2 cells proliferation, inhibited HUVECs and NCI-H446 cells proliferation, decreased HUVECs migration and induced apoptosis of NCI-H446 cells. CONCLUSION: The IVT has a variety of biological activities. This study laid a foundation for the development of novel multi-functional fusion proteins.

Objective To establish carotid artery bifurcation aneurysm in canine, and to clarify whether specific hemodynamic insult in combination with arterial wall degeneration will lead to the occurrence and development of aneurysms. Methods New bifurcation aneurysm model of common carotid artery (CCA) was successfully established in 18 dogs, which were randomly divided into the elastase-treated bifurcation group (EBG,n=9) and the control bifurcation group (CBG,n=9). Three dogs were treated with elastase insult to both straight sections of CCA and were used as elastase-treated straight section group (ESG,n=3). Angiographic and hemodynamic analysis was conducted immediately after the operation, as well as 12 and 24 weeks after the operation. Histopathological examination was performed 12 and 24 weeks after the operation. Results Angiography showed that new aneurysm (mean diameter 3.2 ±0.4 mm) was formed at the apex of CCA bifurcation in 5 dog models of EBG group, while no new aneurysm was observed in both CBG and ESG groups. In EBG group, no rupture of the new aneurysm occurred during the follow-up period. Histological analysis revealed that in EBG group the inner elastic lamina was discontinued, the elastic fiber was disrupted, the muscle layer became thinned, the smooth muscle cells were reduced, and the inflammatory cell (macrophage) infiltration as well as the expressions of MMP-2 and MMP-9 were increased;these changes were statistically significant when compared with those in CBG group and ESG group (P<0.001). Postoperative hemodynamic analysis indicated that in EBG group the wall shear stress at the apex of CCA bifurcation was reduced, the blood flow velocity was decreased, with the relative and total pressure being the highest;all the above changes returned to normal after arterial wall remodeling. Conclusion The results of this study indicate that the arterial wall degeneration and the hemodynamic effect at the apex of CCA bifurcation can lead to new aneurysm formation in canine model.

AIM: To investigate the mechanism by which TA9902 inhibits the formation of amyloid ?-peptide (A?) fibrils. METHODS: Fourier-transform infrared spectroscopy was used to study the secondary structure changes on aging A? in vitro. RESULTS: The content of ?-pleated sheet were 46.53% in the condition of A? aged alone for 30 min. When A? aged alone for 72 h, the content of ?-pleated sheet increased about 19.4% and produced a shift of random coil toward ?-pleated sheet. TA9902 induced a significant decrease in the content of ?-pleated sheet (36.09%). CONCLUSION: TA9902 effectively diminishes the ?-pleated structural content. The effect of TA9902 on the secondary structure of aged A? is associated with inhibition of A? aggregation and fibril formation.

AIM:To gain the molecular mechanism of active ingredients of EGb 761(ginkgo flavones and ginkgo lides) for the inhibition against A?_(1-42) aggregation and fibrogenesis.METHODS:Circular dichroism spectra(CD) and FI-IR were used to curve-fit and analyze the change in A?_(1-42) second structure under A? aged condition and intervention of ginkgo flavones and ginkgo lides.RESULTS:Analysis that was made of 1700-1600 m(-1) amide-Ⅰ-band and curve fitting indicated that after 30 min to 72-incubation,increase of ?-sheet in A?_(1-42) by(18.5%),but the reduction of ?-sheet by(50.95%) and(36.09%) in 72 h incubation with ginko flavones and ginkgo lides,respectively.And increase of ?-turn by(56.56%) and(46.56%) in sane condition.CONCLUSION:Obviously,?-sheet turned into ?-turn.-CH_2 and-CH groups of ketones and alkanes engaged in A?_(1-42) molecular change.

AIM: To observe the humoral immune response in adult rhesus monkey induced by A? 1-15 vaccine. METHODS: 5 adult male rhesus monkeys were injected intramuscularly with A? 1-15 vac cine at baseline and at week 2, 6, 10, 14, 18, 22. The titer and IgG isotypes of the antibody against A? 1-42 in the serum were measured with ELISA. The specificity of the antibody against A? 1-42 was determined by Wester n blotting. The A? plaques in Tg2576 transgenic mouse brain were stained with t he antisera using immunohistochemistry method. RESULTS: At the eighth week after the vaccination, antibody against A? 1-42 bega n to develop significantly i n the serum. The titers of the antibody increased following vaccine boosted and reached 1: 3 840 at the twenty-fourth week, then decreased after the terminat ion o f inocunation. The IgG1 was accounted for the highest level in the antisera pool . The antibody against A? 1-42 showed high specificity. The A? plaques in Tg2576 transgenic mouse brain were labeled with the antisera. CONCLUSION: A? 1-15 vacci ne could induce vigorously specific humoral immune responses in adult rhesus mon key.

AIM: To clarify if TA9901, a natural antioxidants, could inhibit the formation of ?-amyloid(A?) fibril when A? 1-40 were injected into cerebral cortex of rat brain, and explore the mechanism of action of TA9901 on Alzheimer disesse. METHODS: Twelve Wistar rats (250-300 g) were randomly divided into four groups ( n=3 ). (1) control group; (2) TA9901 treatment group (ip. 100 mg?kg -1 ?d -1 ); (3) Vitamin E(VE) treatment group (ip. 100 mg?kg -1 ?d -1 ); (4) PBS group. 5 ?L 0.2% A? 1-40 was immediately injected into the right side of the deep cerebral cortex of control, TA9901 and VE group rats. The animals were sacrificed at the seventh day after the injection. The sections of the rat brain that contained the injected field were examined with transmission electron microscopy and Congo red staining with polarized microscopy. RESULTS: Many depositions of high electron density were observed by electron microscopy in the field where A? 1-40 was injected. They are intimately intermingled with macrophages and astrocytes. In the field, about 10nm fibrillar structures were observed that appeared similar to the fibrils seen in senile plaque (SP) of the brain of Alzheimer disease (AD). The fields in control and VE group contained richer A? fibrils than that in TA9901 group. After the sections stained with Congo red, A? 1-40 aggregation demonstrated intense birefringence under, indication the formation of amyloid fibrils. In TA9901 group, there was a weak birefringence.CONCLUSIONS: TA9901 can inhibit the fibril formation of A? that was injected into deep cerebral cortex of rat brain, this indicates primarily that TA9901 may be a potential therapeutic drug to interfere with the progression of amyloidgenesis in AD.

Circular RNA (circRNA) is a non-coding closed-loop single-stranded RNA molecule lacking the 5' end cap and the 3' poly (A) tail. Circular RNA is more abundant and stable than linear mRNA, and its expression is more conservative and specific. circRNA regulates cancer development through a variety of mechanisms, including miRNA sponges, regulating gene transcription, regulating RNA-binding proteins, and protein translation. This review summarizes the role of circRNA in cancer and helps to develop new clinical diagnostic techniques and treatments.

Objective To investigate the diagnostic accuracy of contrast-free three dimensional time-of-flight (3D-TOF-MRA) with VR at 3.0 T in the detection of intracranial aneurysms in a large cohort of patients prospectively.Methods Four hundred and eleven patients with suspected aneurysms and other cerebral vascular diseases received contrast-free 3D-TOF-MRA examinations at 3.0 T MRA 2 weeks prior to DSA examination.2D-DSA and VR-DSA were regarded as the gold standard.Six patients were excluded because of motion artifacts,and 36 patients were excluded due to lack of VR-DSA data.Accuracy,sensitivity an specificity in detecting intracranial aneurysms were determined by patient-,aneurysm-,and aneurysm sizebased (＜ 3 mm,3-5 mm,＞ 5-10 mm,＞ 10 mm) evaluations.Results In 369 enrolled patients,VR-DSA revealed 306 aneurysms in 246 patients (66.7％) and no aneurysm in 123 patients; VR 3D-TOF-MRA revealed 311 aneurysms in 249 patients and no aneurysm in 120 patients.The patient-based evaluation of VR 3 D-TOF-MRA at 3.0T yielded accuracy of 97.6％ (360/369),sensitivity of 99.2％ (242/244),and specificity of 94.4％ (118/125) in the detection of intracranial aneurysms.The aneurysm-based evaluation yielded accuracy of 98.3％ (524/533),sensitivity of 99.3％ (304/306),and specificity of 96.1％(220/229).The evaluation based on aneurysm sizes (＜ 3 mm) yielded accuracy of 96.4％ (214/222),sensitivity of 98.2％ (112/114),and specificity of 94.4％ (102/108).Conclusion VR 3D-TOF-MRA at 3.0 T MR can detect intracranial aneurysms accurately and may replace DSA as a contrast-free,noninvasive and non-radiation-based modality for the diagnosis and screening of intracranial aneurysms.

BACKGROUND: It has been demonstrated that amyloid-beta 42 protein (Aβ42) immunization in transgenic mouse models of Alzheimer disease(AD)can induce specific Aβ42 antibody, clear Aβ from the brain, and thereby improve spatial learning and memory. It has been a promising treatment strategy for AD.OBJECTIVE: To explore the effect of Aβ42 and its subunit vaccines immunization on spatial learning and memory of APPSWE transgenic mice.DESIGN: A randomized controlled experiment with mice as subjects.SETTING: The brain research laboratory of the anatomy department in a the medical college of a univeristy.MATERIALS: The experiment was conducted in the Experimental Animal Center and the Anatomy Department of Sun Yat-sen University from April 2003 to February 2004. Thirty-two APPSWE transgenic mice of 5 months old were bought from Taconic Company, USA. The second generation of mice were successfully reproduced in the Anatomy Department. These mice were randomly assigned into four groups: control group, Aβ42 group, Aβ1-15group, and Aβ36-42 group. Each group contained 8 in each group.INTERVENTIONS: Aβ42 and its subunits combined with MF59 adjuvant were subcutaneously injected for fundamental immunity and then applied in nasal mucosa for intensified immunization. The immunization period was 8 months. Y-maze was used for behavior test before immunization and Morris water maze was used after immunization.MAIN OUTCOME MEASURES: Spatial learning and memory, mean escape latency, times of passing through the platform point, swimming distance percentage of the first quadrant, and swimming distance percentage of the 20% marginal area.RESULTS: The correct reaction times in Y-maze behavior test were 7.50 ±0. 81, 7.06 ±0.71, 7.19 ±0.91, and 7.50 ±0.86 respectively in the control, Aβ42, Aβ1-15, Aβ36-42 groups and there was no significant difference ( P ＞ 0. 05) . After immunization, the mean escape latencies in 8 units of localized navigation test were(67.3 ±2. 8) s, (23.6 ± 1.6) s, (26.4 ±2.0) s,and (36.5 ± 2.2) s. The results in three experiment groups were different from that in control group and there was no difference between the three experiment groups ( P ＞ 0. 05 ) . The mean times of passing through the platform point in the 4 groups were 0.71 ±0.29, 8.14 ± 1.37, 7.28 ± 1.34,and 3.29 ± 0. 67. Swimming distance percentage of the first quadrant in the4 groups were(24.3 ±2.9)%, (50.6±11.6)%, (49.9±9.3) %,and(35.4±7.0)% and the swimming distance percentages of 20%marginal area were (46.4 ± 7.3 ) %, ( 11.6 ± 3.9) %, ( 14.4 ± 2. 6) %, and (25.8 ± 3.3)%. The mice in three experiment groups showed increase in the times of passing through platform point, swimming distance percentage of the first quadrant, and decrease in distance percentage of 20% marginal area compared with control group. The results in three experiment groups were no significantly different( P ＜ 0. 05).CONCLUSION: Immunization with A342 and its subunits can effectively ameliorate impairment of spatial learning and memory in APPSWE transgenic mice.

Objective To evaluate the efficacy and mid-term follow-up results of endovascular treatment with Willis covered stent for traumatic pseudoaneurysms located in the internal carotid artery (ICA).Methods ICA angiogmphy was performed in 38 patients with traumatic brain and neck injury.Of the 38 patients.13 delayed traumatic pseudoaneurysms were found.All the pseudoaneurysms were treated with Willis covered stents.Follow-up angiography was performed at 1,3,6 and 12 months after the procedure,and the results were categorized as complete or incomplete occlusion.Clinical manifestations were graded as full recovery,improvement,unchanged and aggravation.Results Willis covered stent placement was technically successful in all traumatic pseudoaneurysms.No procedure-related complications occurred.The initial angiographic results showed a complete occlusion in 9 patients,and an incomplete occlusion in 4.The angiographic follow-up within 3-12 months exhibited a complete occlusion in 12 patients and the parent arteries remained patency in all patients.The clinical follow-up observation demonstrated that full recovery wag obtained in 11 patients,clinical improvement in one,and unchanged condition in one.No morbidity or mortality occurred.Conclusion Willis covered stent implantation iS a feasible and practical treatment for traumatic pseudoaneurysms located in the ICA.This technique can well preserve the parent artery with excellent therapeutic results.