Carbon source plays an important role in the constitutive expression of foreign proteins in Pichia pastoris. In present study, glucose , glycerol , methanol and oil acid, was used respectively as the only carbon source to constitutively express hAS in Pichia pastoris GS115 (pGAP9K-AS)in shaking flask. The result shows that oleic acid is the best (163 mg/L) compared with glycerol (83mg/L), glucose (76 mg/L)and methanol (57 mg/L). Since oleic acid is insoluble in water, glycerol was used as the carbon source in the high-density cell culture of GS115 (pGAP9K-AS) in a 30 liter bioreactor and 169 mg/L of angiostatin was obtained after 48h of culture. The expressed angiostatin is immunologically active as shown by Western blotting. The recombinant hAS inhibits bFGF induced CAM angiogenesis and suppresses the growth of B16 melanoma in C57BL/6J mice. The tumor inhibition rate is 90% after 12 days of treatment. Statistics analysis revealed that the tumor volume difference of mice between the hAS group and PBS group is prominent (P < 0.01).
Angiogenesis Inhibitors ; biosynthesis ; genetics ; therapeutic use ; Angiostatins ; biosynthesis ; genetics ; therapeutic use ; Animals ; Bioreactors ; microbiology ; Culture Media ; pharmacology ; Fermentation ; Glycerol ; pharmacology ; Humans ; Melanoma, Experimental ; drug therapy ; Mice ; Mice, Inbred C57BL ; Pichia ; genetics ; growth & development ; metabolism ; Recombinant Proteins ; biosynthesis ; genetics ; therapeutic use

OBJECTIVETo discuss the application of MRI in indirect temporomandibular joint injury without condylar fracture.

METHODSMRI examination on temporomandibular joint was conducted in 28 patients with indirect injury to temporomandibular joint without condylar fracture. The scanning sequence included T(1)WI, PDWI on oblique sagittal section at both open and closed mouth positions, and T(1)WI, T(2)WI on oblique coronal section. The MRI appearance was analyzed by 2 senior radiologists.

RESULTSAmong the 56 temporomandibular joints of 28 patients, 35 joints exhibited pathological changes on MRI, in which there were 9 bone injuries, 21 articular disc dislocation, 24 intracapsular hematocele and hydrops.

CONCLUSIONSMRI can clearly reveal bone injury, articular disc dislocation as well as articular capsule abnormality in the indirect injury of temporomandibular joint without condylar fracture. It is highly advocated in clinical use.

Adolescent ; Adult ; Aged ; Child ; Female ; Humans ; Magnetic Resonance Imaging ; Male ; Maxillofacial Injuries ; diagnosis ; Middle Aged ; Temporomandibular Joint ; injuries