Sphingolipids are structural components of cellular membranes.Their metabolites,such as ceramide,sphingosine,and sphingosine 1-phosphate are important bioactive molecules,which act as the first or second messengers regulating various cellular activities,including proliferation,survival,migration and neovascularization in vitro and in vivo tumor models.Sphingosine kinases 1(SPK1) is a critical regulator of the balance between ceramide and S1P.Recent studies show that ceramide,SPK1 and S1P can regulate many of the hallmarks of cancer.These studies suggest that these metabolites can serve as novel targets for cancer therapy.

Adolescent ; Adult ; Anemia, Aplastic ; blood ; Case-Control Studies ; Female ; Humans ; Isoenzymes ; metabolism ; Leukocytes, Mononuclear ; metabolism ; Male ; Middle Aged ; Protein Kinase C ; metabolism ; Protein Kinase C-theta ; T-Lymphocytes, Cytotoxic ; cytology ; Th1 Cells ; cytology ; Young Adult

According to the existing Provisions for Drug Registration (SFDA Order No. 28), applications for new drugs of traditional Chinese medicine are divided into two parts: the applications for drug clinical trial and for drug production (including new drug certificate). It will last for about 10 years from the application for drug clinical trial to get approving, and it also remains many problems and the low probability to succeed. From the sight of pharmaceutical review, there are mainly two aspects of regulatory compliance and technical issues, mainly for changes without approval of the competent authorities of the country. For example, sample preparation and approval of clinical trial process are significant changes. Technical problems are reporting incomplete data or information submitted does not comply with the technical requirements for review, such as: production process validation does not provide information, the preparation of samples for clinical trials and field inspection, production information, or the information provided does not meet the technical requirements. This paper summarizes the frequently asked questions and to make recommendations to advise applicants concerned, timely detection of problems, avoid risk, improving the quality and efficiency of the application for registration.
China ; Drug Approval ; legislation & jurisprudence ; Drug Evaluation ; legislation & jurisprudence ; Humans ; Legislation, Drug ; Medicine, Chinese Traditional

Objective To construct pcDNA3-HIF-1? eukaryotic expression vector and to investigate its function in breast cancer MCF-7 cells.Methods RT-PCR was applied to amplify human HIF-1? cDNA from MCF-7 cells with a pair of sequence specific primers carrying a restriction enzyme site XbaⅠ or HindⅢ on each 5′end.HIF-1?(cDNA) was inserted into pMD18-T after sequencing,and then inserted into pcDNA3 vector.The recombinant vector was transfected into MCF-7 cells to observe its expression and function by Western blot and dual-luciferase reporter gene assay.Realtime-PCR was performed to detect the inducible expression of C-MET mRNA by HIF-1?,the anti-apoptotic effect of HIF-1 under hypoxia was analysed by detecting the Caspase3/7 activity.Results The sequence of pcDNA3-HIF-1? is identical with the gene bank.It enhances the expression of HRE reporter gene and C-MET mRNA,but decreases the Caspase3/7 activity in MCF-7 cells under hypoxia.Conclusion The pcDNA3-HIF-1? eukaryotic expression vector was successfully constructed,it not only has strong DNA binding and inducing activity,but also has anti-apoptotic effect in hypoxia MCF-7 cells.

Objective To construct pGL3-EPO-HRE reporter gene vector and to detect its function.Methods To synthesize two single-strand DNA fragments which had overlap sequence at the 3'end,then mixed them to construct the double-strand DNA fragment which include three copies of(57 bp) human erythropoietin(EPO) HRE.There was MluⅠor BglⅡ digestive site at the end of this DNA fragment.After inserting the(87 bp) fragment into pMD18-T for sequencing,the correct sequence was inserted into pGL3-promoter to construct pGL3-EPO-HRE.The recombinant vector was co-transfected into MCF-7 cells with pcDNA3-HIF-1?or pcDNA3 respectively,(12 h) later cells were treated with(100 ?mol/L) cobalt chloride(CoCl_(2)) for(24 h), then the expression of pGL3-EPO-HRE was observed by dual-luciferase reporter gene assay method.Results The sequence of pGL3-EPO-HRE was identical with human EPO HRE sequence.pGL3-EPO-HRE luciferase activity increased about 10 fold(P

Aged ; Humans ; Male ; Microbial Sensitivity Tests ; Middle Aged ; Pneumoconiosis ; drug therapy ; microbiology ; Tuberculosis, Multidrug-Resistant ; drug therapy ; microbiology

Anti-Bacterial Agents ; therapeutic use ; Bartonella henselae ; Cat-Scratch Disease ; drug therapy ; microbiology ; physiopathology ; Child ; Female ; Humans

AIM: To study the preconditioning effect of nitrate medicine on different parts of the heart and its signaling pathway. METHODS: The cells from different parts of the heart (atrium, left ventricle, right ventricle, apex and the whole heart) were isolated and cultured. The cultured cells were treated with hypoxia and reperfusion, hypoxia preconditioning, 1 mmol/L L-arginine preconditioning and 5 mmol/L L-arginine preconditioning, respectively. The cell necrosis rate, cell apoptosis rate, LDH concentration in the medium, the [Ca 2+]i and PKC activity in the cells were determined. RESULTS: The result show that hypoxia and reperfusion increased the necrosis rate and apoptosis rate, enhanced the concentration of LDH in the medium and the [Ca 2+]i overload in the cells (P0.05). After hypoxia preconditioning and L-arginine preconditioning, these indexes decreased (P

Objective To observe the expression of c-fos mRNA and protein in fluoride treated mouse fibroblast (FB) and osteoblast (OB) and to further explore the effects of c-fos in the osteogenic action of FB. Methods Mouse FB and OB were divided into control group and six fluoride groups (0, 0.0001, 0.0010, 0.1000, 1.0000, 10.0000,20.0000 mg/L F-), and the levels of c-fos protein at 2,4,24,48,72 h and c-fos mRNA at 48 h were measured by using ELISA and RT-PCR methods. Results Compared with the control group, fluoride increased the content of c-fos protein obviously in all FB group(P<0.01); and it is increased in 0.0001,0.0010 mg/L groups at 48 h (0.73±0.04, 0.64±0.14) and 0.0001 mg/L group at 72 h(0.70±0.17) in OB compared with the control group (0.32±0.04,0.27±0.05, P<0.01). Compared with the control group (0.95±0.11), RT-PCR revealed an increasing tendency of the expression of c-fos mRNA at 48 h in FB (1.06±0.16, 1.06±0.12,1.12±0.16,1.04±0.15,1.04±0.10,1.15±0.29), but the difference was not statistically significant(P>0.05); however, a statistically significant difference(P<0.01) of c-fos mRNA in 20.0000 mg/L group(1.40±0.17) in O B was found compared with the control group (1.06±0.06). Conclusion The higher expression of c-fos mRNA and protein in FB induced by fluoride may play an important role in the transformation of osteoblastic phenotype as well as increase the osteogenesis ability in FB.

Objective To study the diagnostic value of multi-direction adjusted multiplanar reconstruction (MPR)by 64-slice CT myelography (CTM)in diagnosing cervical nerve injury,and the possibility of the MPR to replace conventional myelography and CT direct-scanning axial images.Methods Twenty-six patients with cervical nerve root injury were examined by conventional myelograpby and 64-slice CT using isotropic parameters.Then multi-direction MPR were performed to display nerve roots on coronal and sagittal planes besides axial images.Twenty-six patients were performed surgical operations and diagnosis were obtained.The coincident diagnosing rate with surgical operations results were compared statistically among multi-direction MPR,direct-scanning axial CT images,conventional myelography.The numbers of images were also compared between axial MPR and direct scanning axial CT images.Results Direct sign of nerve root avulsion was the loss of normal nerve root defect seen in the Isovist filled thecal sac in 6d-slice CT,which was found in 31 nerve roots.Indirect signs included:(1)Traumatic pseudomeningocele: 29 nerve roots showed the leak of Isovist into nerve root sheath,and extended into foramina;(2)Arachnoid cyst: 26 nerve roots clearly displays cystic distension in nerve root,which has low- density fine clew form septation from subarachnoid cavity and no nerve root in the cyst ;(3)Deformity of the subarachnoid space : deformity of thecal sac,partially lack of Isovist into arachnoid space,which was found in 17 nerve roots.The coincident diagnosing rate of cervical nerve root injury by multi-direction adjusted coronal MPR imaging was 92.6% (50/54),which was higher than by axial CT (77.8%,42/54)and conventional myelography (68.5%,37/54),There was significant difference between the conventional myelography, direct-scanning axial CT,multi-direction MPR images (Kappa = 0.686,0.772,0.920, respectively,P