〔Abstract〕 Based on analyzing the background of generic drugs research and development, the paper elaborates the utilization meth-ods of drugs invalid patents, including information sources optimization and retrieval, invalid patents selection indexes and methods, in-fringement avoidance strategy.It takes the gastrointestinal stromal tumor drugs for example to find out drugs which can be imitated, pros-pects the future application of invalid patents information.

Cells, Cultured ; Endothelial Cells ; cytology ; Humans ; Stem Cells

Acute Disease ; Adult ; Central Nervous System Diseases ; diagnosis ; etiology ; Decompression Sickness ; complications ; diagnosis ; Humans ; Magnetic Resonance Imaging ; Male ; Retrospective Studies ; Young Adult

BACKGROUND: Embryonic stem cells (ESCs), the seed cells of all mature cells in vivo, are useful tools for nerve transplantation and developmental gene function research. Notch1 signaling pathway is the key pathway to control the ordered neural development and differentiation of many kinds of neural cells, however, there is no report on the dynamic expression of Notch1 signal during the ESC differentiation to date. OBJECTIVE: To investigate the expression of Notch1 protein, transmembrane signal transduction molecule, during directional differentiation of embryonic stem cells into neural cells. DESIGN, TIME AND SETTING: Cell research was carried out between October 2003 and October 2004 at Zhongshan Ophthalmic Center, SUN Yat-sen University, Guangzhou, Guangdong Province, China. MATERIALS: BALB/C mouse embryonic stem cell line Ⅵ (passage 11)was obtained from experimental animal center of SUN Yat-sen University, provided by professor Huang Bing. ESC culture medium was high-glucose DMEM medium with 20% bovine serum and 106 IU/L mouse leukemia inhibitory factor. Induced differentiation medium was high-glucose DMEM medium with 20% fetal bovine.serum and 5×107 mol/L retinoid acid(RA). METHODS: Passage 11 ESCs were resuscitated and incubated by ESC culture medium in incubator at 37℃ with 5% CO2. Passage 11 ESCs were subcultured after 2 or 3 days and RA was added into medium to induce differentiation. Three time points for observation were established: induced for 1, 5 and 9 days. MAIN OUTCOME MEASURES: Morphological changes were observed under inverted phase contrast microscope, MAP-2 antigen expressed in differentiated cells was detected by immunofluorescence method. Immunocytochemistry, Western Blot, flow cytometry assay were used to investigate the Notch1 protein expression. RESULTS: ESCs presented clone-like growth. After induced by RA for 9 days, single neural network was achieved around most of the cell clusters. With the prolongation of induction, MAP-2 positive neural cells increased gradually. Almost all ESC clones expressed Notch1 protein strongly or positively, but Notehl protein expression decreased gradually after induced differentiation (P < 0.01). CONCLUSION: Notch1 signal shuts off progressively during induction of ESCs into neural cells, which suggests Notch1 may play an important role in the differentiation of ESCs into neural cells.

Objective To investigate the relationship of the serum levels of high sensitivity C-reactive protein(hs-CRP) and homocysteine with middle cerebral artery(MCA) intraplaque hemorrhage.Methods A total of 63 patients who met the inclusion criteria showed high-grade(≥ 70％)MCA stenosis.The levels of serum hs-CRP and Hcy were detected.All patients were divided into symptomatic group and asymptomatic group,intraplaque hemorrhage group and non-intraplaque hemorrhage group.All patients performed conventional MRI and high-resolution MRI(HR-MRI).The correlation of the serum levels of hs-CRP and homocysteine with middle cerebral artery intraplaque hemorrhage was analyzed.Results Totally 37 symptomatic and 26 asymptomatic MCA stenoses were analyzed.The occurrence rate of intraplaque hemorrhage was significantly higher in symptomatic MCA stenosis group than in asymptomatic MCA stenosis group (40.5 ％ vs.11.5 ％,x2 =6.29,P＜ 0.05).Symptomatic group displayed a higher hs-CRP and Hey levels (8.97 ± 3.36 mg/L and 20.00 ± 3.16 μmol/L,respectively) than did asymptomatic group [(5.26 ± 3.12) mg/L and (12.22 ± 1.88) μmol/L,t =4.43 and 11.23,respectively,each P ＜ 0.001].The hs-CRP and Hcy levels were higher in intraplaque hemorrhage group [(10.53 ± 3.59) mg/L and (21.70 ± 2.40) μmol/L,respectively] than in non-intraplaque hemorrhage group[(6.20 ± 3.02) mg/L and(11.77±1.69) μmol/L,t=4.87 and 18.58,respectively,each P＜0.001].The hs-CRP and Hcy levels were positively correlated with the risk for middle cerebral artery intraplaque hemorrhage(r=0.461 and 0.519,each P＜0.001).Conclusions The serum levels of hs-CRP and Hcy are closely associated with middle cerebral artery intraplaque hemorrhage,which can be used to evaluate the stability of plaque.The MCA intraplaque hemorrhage is associated with ipsilateral stroke.

Objective: To observe the effect of phenylephrine (PE) on pressure overload induced myocardial ifbrosis (MF) with its relevance to transforming growth factor-β1 (TGF-β1), drosophila mothers against decapentaplegic protein 3 (smad3) and connective tissue growth factor (CTGF) signal pathway in experimental rats.
Methods: A total of 28 male SD rats were randomly divided into 4 groups: Control group, AAC (abdominal aorta coarctation) group, AAC+PE group and AAC+prazosin group.n=7 in each group. Collagen volume fraction (CVF) of left ventricle was observed by myocardial collagen morphology, left ventricular myocardial tissue protein expressions of α-smooth muscle actin (α-SMA), TGF-β1, smad3 and CTGF were measured by immunohistochemistry, protein expression of α-SMA was also examined by Western blot analysis.
Results:①Myocardial collagen morphology presented that compared with Control group, AAC, AAC+PE and AAC+prazosin groups had increased CVF, allP<0.01; compared with AAC group, AAC+PE group had decreased CVF, P<0.01.②Immunohistochemistry demonstrated that compared with Control group, AAC, AAC+PE and AAC+prazosin groups had up-regulated protein expressions of α-SMA, TGF-β1, smad3 and CTGF, allP<0.01; compared with AAC group, AAC+PE group had down-regulated protein expressions of α-SMA, TGF-β1, smad3 and CTGF, allP<0.01.③Western blot analysis indicated that compared with Control group, AAC, AAC+PE and AAC+prazosin groups had the higher α-SMA expression, allP<0.05; compared with AAC group, AAC+PE group had the lower α-SMA expression, P<0.01.
Conclusion: Phenylephrine could improve pressure overload induced MF in experimental rats which might be related to TGF-β1/smads signal pathway inhibition and CTGF down-regulation.

OBJECTIVE:To study the chemical constituents in antioxidant activity part of Semiaquilegia adoxoides. METH-ODS:The antioxidant activity part of S. adoxoides were isolated and purified by chromatography on silicagel and Sephadex LH-20 column,and compound structures were identified physicochemical properties and spectral data analysis. RESULTS:Nine com-pounds were isolated from the antioxidant activity part of S. adoxoides,namely magnoflorine (1),griffonilide (2),salidroside (3),ferulic acid(4),genistein(5),2,4-dihydroxybenzoic acid(6),chlorogenic acid(7),caffeic acid(8)and p-coumaric acid (9). CONCLUSIONS:The study confirms the active material basis in antioxidant activity part of S. adoxoides for the first time.

Objective To investigate the relationship between plasminogen activator inhibitor-1(PAI-1) activity and PAI-1 promoter 4G/5G polymorphism in patients with type 2 diabetes from the coastal areas of Shandong province.Methods The 4G/5G allele polymorphism in the PAI1 gene promoter region were tested by allele specific PCR in 116 type 2 diabetes and 40 normal controls.The activity of plasma PAI-1 was assayed by chromogenic substrate method.Results The plasma PAI-1 activity in patients was higher than that in controls(P

Objective: To investigate the inhibitory effect of Dahuangzhechong pill to platelet activation.Methods: The blood platelet were incubated with the medicated blood plasma with Dahuangzhechong pill,and then activated with ADP and marked with PAC-1 monoclonal antibody.The activation rate of blood platelet was analyzed by flow cytometer.The patients with coronary heart disease or cerebral infarction took Dahuangzhechong pill,after one course of treatment,the patients,blood platelet were separated and then incubated with PAC-1 monoclonal antibody.The activation of blood platelet was detected by ? ow cytometer.Results: Compared with aspirin group(51.7%),the activation rate(20.82%) of blood platelet in Dahuangzhechong pill group decreased(P

Objective To investigate the infection and distribution of human papilloma virus (HPV) types in patients with infiltrative cervical cancer (ICC) and cervical intraepithelial neoplasia (CIN) in Chongqing. Methods Fifty-eight cases diagnosed with ICC or CIN[KG-*6]Ⅱ to Ⅲ,which were confirmed by histopathology in the Tumor Hospital of Chongqing,were included in the analysis. DNA enzyme immunoassay (DEIA) and SPF10 LiPA25 (version 1) testing was used for DNA extraction and extension,and linear probe reverse hybridization to detect lesions HPV type. Results HPV infection were detected in 56 of 58 specimen,the positive rates of CINⅡ,CINⅢ and cervical cancer were 100.00%,100.00% and 95.00% respectively. A total of 13 types were detected. HPV16 infection rate was up to 60.00%,followed by HPV18(7.69%),HPV58(7.69%),and HPV31(4.62%). Conclusion HPV16 is still the most common type in Chongqing. HPV18,HPV58 and HPV31 also has a high prevalence.