[Treatment of non-traumatic femoral head avascular necrosis by perfusion of bone marrow stromal stem cells through optional artery].

OBJECTIVE: To study the medium and long term effects of perfusion of bone marrow stromal stem cells through optional artery for the treatment of non-traumatic femoral head avascular necrosis. METHODS: From January 2000 to December 2004,62 cases(78 hips) with non-traumatic femoral head necrosis accepted optional artery marrow stromal stem cells infusion treatment and had complete follow-up data, including 43 hips of 35 males and 35 hips of 27 females with an average age of 36.3 years old (22 to 54). According to preoperative imaging data, 16 hips were ARCO I stage, 52 hips were II stage, 10 hips were III a stage. Harris score was 64.94 +/- 8.12 preoperatively. Postoperative Harris score at the last follow-up, imaging changes,DSA vascular changes were analysis. RESULTS: The patients were followed up for 9 to 13 years (means 11 years). By the end of the follow-up, a total of 18 hips got artificial joint replacement, 10 hips of preoperative ARCO I, II period got artificial hip joint replacement, 8 hips of IIIa period got hip artificial joint replacement. Harris score was 71.21 +/- 0.19 at the end of the follow-up, it was obviously enhanced compared with preoperative. DSA showed blood vessels of supply the femoral head increased thickening. CONCLUSION: Perfusion of bone marrow stromal stem cells through optional artery can effective treat non-traumatic femoral head necrosis of ARCO I, II period, it can make the femoral circumflex artery and its branches increased thickening.

Establishment of stable cell line and expression and purification of HIV gp120 in Drosophila S2 cells / 病毒学报

Constructing eukaryotic expressing vector of pMT/Bip/V5-His A-HIV gp120 and transfecting S2 cells to establish stable gp120-expressing S2 cell line. The gp120 gene of HIV CRF07-BC epidemic in China was amplified by polymerase chain reaction from the recombinant vector PRSV-gp120 and confirmed by DNA sequence analysis. The DNA fragment of HIV gp120 was digested with the restriction endonucleases NcoI and XhoI, then inserted into eukaryotic expressing vector pMT/BiP/V5-His A. A selection vector containing the streptomyces griseochromogenes bsd gene conferring blasticidin resistance was cotransfected into drosophila S2 cells by Cellfectin II reagent. The stable cell line was established following repeated screening by blasticidin. HIV gp120 was purified by a Ni-NTA affinity column followed by molecular sieve chromatography. Recombinant protein was characterized by SDS-PAGE, Western blot and ELISA. The eukaryotic expressing vector pMT/BiP/V5-His A-HIV gp120 was constructed, stable expressing cell line was established, and protein was expressed and purified successfully. This result will contribute to functional study of gp120 and vaccine design against AIDS.