0.05)]. Conclusion The elevated serum HA is linked to a potential marker in kidney transplantation.

BACKGROUND: Previous studies have demonstrated that LRP16 is an estrogen-responsive gene. Its expression level is strongly associated with the proliferation and invasive growth of human breast cancer cells.OBJECTIVE: To construct a LRP16 targeting vector and screen mouse embryonic stem cell clones with homolougous recombination of an inactive LRP16 gene.DESIGN: Constructing an inserting inactivation target by inserting SA-RIES-β geo expression cassette.SETTING: Bioregulatory Laboratory of the Third Medical Department of Kyushu University in Japan and Department of Molecular Biology, General Hospital of Chinese PLA.MATERIALS: The materials used here were mainly provided by the Bioregulatory Laboratory, the Third Medical Department of Kyushu University in Japan. The mouse genomic library in pBeloBAC11 Vector was purchased from lnvitrogen Corp. The competent TopF10 was purchased from Beijing Tiangen Biotech Corp. pcDNA3.1(+) vector was kept in our laboratory. Mouse ES cells were provided by Kyushu University.METHODS: The experiment was performed in Kyushu University and Department of Molecular Biology of PLA General Hospital from November 2004 to May 2005. Targeting sequence of LRP16 gene was obtained from 129 mouse genomic Bacterial Artificial Chromosomes library based on polymerase chain reaction (PCR) screening. The SA-RIES-β geo fragment was inserted within LRP16 fifth exon to inactivate LRP16. ES cells were screened with G418 and the homologously recombinant clone was identified by Southern blot analysis.MAIN OUTCOME MEASURES: Clones with homologous recombination.RESULTS: The LRP16 fragment including exon 5 to 11 was subcloned into the pBluescript SK Ⅱvector. Restriction map demonstrated that the SA-IRES-β geo fragment was correctly inserted into the LRP16 fifth exon. Southern blot results showed that there was an ES clone with targeting sequence homologously inserted.CONCLUSION: A LRP16 gene targeting vector is constructed and a homologous recombinant is obtained.

Clinical and imageological characteristics of Paget's disease of bone in 7 patients who were treated in our hospital from 1991 to 2007 were analyzed. The data showed (1) 6 patients had the symptoms of bone pain and bone deformity, with more long bones involved; (2) Serum alkaline phosphatase was over normal range; (3) Imageology showed that osteoclasia was usually combined with bone sclerosis; (4) All the patients were treated with bisphosphonates; (5) 2 patients with serious bone deformity were treated with orthomorphia. Paget's disease of bone is a kind of metabolic bone disease with the characteristics of osteoclasia combined with bone sclerosis. The main features are bone pain and bone abnormality. Bisphosphonates are the first choice of drugs. Patients with serious bone deformity should be treated with orthomorphia.

Objective To predict the CTL epitopes of Tat exon 1 region in HIV-1 CRF07_BC strains, which were prevailing in China. Methods Total of 236 plasma samples were from the 3rd National HIV Molecular Epidemic Survey (NMES3). All the subjects were infected with HIV-1 CRF07_BC viruses. The tat exon 1 region was amplified by reverse transcription reaction and nested polymerase chain reaction (nested-PCR), then the PCR products were sequenced. The distribution of CTL epitopes of this region were predicted by on-line software BIMAS HLA Peptide Binding Predictions and statistics software. Results To-tal of 236 CRF07_BC strains were from 16 provinces, mainly in intravenous drug asers(58.9%)and then sex(25.0%). It was showed that there were 12 CTL epitopes of 236 Tat exon 1 region of CRF07_BC strains mainly located in proline-rich region, cysteine-rich region and core-region. Those epitopes were banded by 5 HLA presenting molecules in genotype(A * 2501 ,A * 2902, B * 15,B * 5301 and Cw * 1203) and 6 HLA presenting molecules in serotype (B53, B58 ,B57 ,A3 ,A68 and Cw12). The frequency of single amino acid substitution was more than 50% in 7 CTL epitopes. Conclusion The CTL epitopes in Tat exon 1 of CRF07 _BC strains were located in different functional regions, and there were some amino acid variations in them.

BACKGROUND: Artificial humeral head replacement is an effective method for the treatment of complex proximal humeral fractures, which has received good results in relieving pain. However, the final functional recovery is unpredictable. OBJECTIVE: To compare biomeshanical stability between anatomical and overlapping reconstruction of the greater tuberosity in cadaveric humeral head replacement models.METHODS: Eight pairs of fresh-frozen shoulder cadavers (16 shoulder joints) were match-paired into two groups. Standardized humeral head replacement procedure was performed in all specimens, and anatomical and overlapping reconstruction of thegreater tuberosity was adopted in each group respectively. For overlapping group, the greater tuberosity was reattached to the proximal humeral shaft in an overlapping style, which was achieved by an additional 5 mm bone osteotomized from the medial cortex of the humeral diaphysis. Custom mounting apparatus and fixation jigs were designed for designated shoulder motion.RESULTS AND CONCLUSION: When the shoulder was external rotated to neutral position, the mean displacement of greater tuberosity in the anatomical reconstruction group was smaller than that of the overlapping reconstruction group (P < 0.05). When the gleno-humeral joint was elevated to 30～ and 60～ forward flexion (accounting for 45° and 90° shoulder forward flexion), there was no significant difference of greater tuberosity displacement between the anatomical group and overlapping group. The findings demonstrated that, although overlapping reconstruction can increase the bone healing area between the greater tuberosity and the humeral diaphysis, there may be some loss in mechanical stability as the trade-off. Even though we strictly follow the standardized postoperative rehabilitation protocol after humeral head replacement, prominent displacement between the greater tuberosity relative to the humeral diaphysis was detected. Accordingly, postponing of the postoperative rehabilitation program after humeral head replacement for a decent period may improve tuberosity healing.

Objective To summary the pathological and genetic features in nine Chinese limb girdle muscular dystrophy 2I (LGMD2I) patients.Methods Nine LGMD2I patients were recruited from Peking University First Hospital between 2011 and 2016, who came from nine unrelated and non-consanguineous families.The mean age of onset was (8.2±5.2) years (2 to 19 years), and the mean disease duration was (10.4±6.1) years (1 to 24 years).There were six males and three females, present with weakness in limb girdle muscles, four of whom accompanied with calf hypertrophy and three with scapular winging.Serum creatine kinase was 964-23 131 U/L (normal 25-190 U/L).Five of them who conducted electromyogram showed myogenic pattern.Muscle biopsy and next generation sequencing were performed in these patients, then sanger sequencing was performed to determine whether the variants co-segregated with the phenotype in these families.Results Muscle biopsy revealed myopathy features in six patients, dystrophic change in one, and only mild changes in two.Major histocompatibility complex-Ⅰ was positive in six cases, and rimmed vacuoles were found in two.There were seven mutations in fukutin-related protein (FKRP) gene.A reported mutation of c.545A>G (p.Y182C) appeared in eight cases, including three homozygotes and five compound heterozygotes.The mutation of c.1067T>C (p.Ile356Thr) was reported too.And c.1263C>A (p.Tyr421X), c.534G>T(p.Thr178Cys), c.1027G>C (p.Glu343Gln), c.1027G>T(p.Glu343X), c.1448A>G (p.Tyr483Cys) were found to be novel mutations.Conclusions LGMD2I showed large variation in myopathology.The missense mutation A545G(Y182C) is a hot spot of FKRP gene in our series.

Objective To investigate the correlation of the 24?hour ambulatory systolic blood pressure (SBP) and carotid intima?media thickness(CIMT) in the elderly. Methods A total of 2 464 who were more than or equal to 60 years old participants were selected with random sampling in accordance with the inclusion criteria from the retired workers in Tangshan Kailuan Company. Dynamic blood pressure monitoring, neck vascular ultrasound and other examination were performed for the participants. . Multivariable linear regression analysis was used to analyze correlation between the SBP of 24?hour, daytime and nightime with CIMT, respectively. Results ( 1) The observation population was divided into three groups according to the tertiles of SBP of 24?hour, daytime and nightime, respectively. With the levels of different SBPs inceasing, CIMT values thickened markedly ( P<0. 01 ) . ( 2 ) Multivariable linear regression analysis showed that after adjusting for confounding factors,the SBP of 24?hour,daytime and nightime associated with CIMT positively and linearly(P<0. 05),and regression coefficient(95%CI) were 0. 022(0. 009-0. 035), 0. 021(0. 008-0. 035), 0. 019 ( 0. 006-0. 032) respectively. In addition,clinic SBP step into the multivariable linear regression,and regression coefficient ( 95%CI ) were 0. 016 ( 0. 003-0. 029 ) , 0. 016 ( 0. 003-0. 030 ) , 0. 019 ( 0. 007-0. 032 ) , respectively. Conclusion The effect of increased 24?hour ambulatory SBP on CIMT was greater than the clinic SBP. Active monitoring of 24 h ambulatory blood pressure and maintaining a low level of blood pressure is essential for preventing and delaying atherosclerosis.

Objective To analyze the antibody responses in guinea pigs vaccinated with recombi-nant vaccinia virus( rTT) strains expressing transmitted/founder ( T/F) HIV-1 membrane proteins in combi-nation with gp140 protein.Methods Guinea pigs were primed with rTT strains and boosted twice with gp140 protein in every four weeks.Serum samples were collected from guinea pigs before immunization and in 2, 6 and 10 weeks after the last immunization for the detection of HIV-1-specific binding antibodies, neu-tralizingantibodiesandtherelativeavidityofantibodies.Results (1)Thebindingantibodiesspecificto HIV-1 B′/C, B, AE subtypes were efficiently induced by the immunization of rTT-B, rTT-C and rTT-CON vaccinia strains in combination with gp140 protein.The antibody titers ranged from 111 430 to 1 024 000. More antibodies against HIV-1 B′/C and AE subtypes were induced in guinea pigs by the immunization of rTT-C and rTT-CON strains in combination with gp140 protein than those by using rTT-B strain prime-protein boost strategy (P<0.05).No significant differences with the titers of HIV-1 B subtype specific antibody were observed among the guinea pigs immunized with the three strategies.( 2 ) High titers of SF162 and ZM109 neutralizing antibodies were induced in guinea pigs immunized with rTT-B, rTT-C and rTT-CON vac-cinia strains in combination with gp140 protein, ranging from 83.76 to 649.30.No significant differences were found among the three groups.(3) The HIV-1 V1V2-gp70 specific antibodies associated with protec-tive immunity were induced by immunization of the three virus prime-protein boost strategies.No significant differences were observed among them.(4) Antibodies induced in guinea pigs by immunization of the three strategies showed strong affinity to membrane proteins of HIV-1 B′/C, B, AE subtype strains.No significant differences were found among the three immunization strategies.Conclusion A strong humoral immune re-sponse was induced in guinea pigs primed with recombinant vaccinia virus strains expressing T/F virus HIV-1 membrane proteins and boosted with gp140 protein.

OBJECTIVETo explore the relationship between NB4 cell apoptosis induced by tanshinone IIA (TanIIA) and the cell mitochondrial transmembrane potential (DeltaPsim).

METHODSNB4 cells were treated with TanIIA, As(2)O(3), TanIIA plus 1.0 micro g/ml CsA and As(2)O(3) plus 1.0 micro g/ml CsA, respectively. Morphological changes were observed under light microscope and transmission electron microscope. The percentages of sub-G(1) cells and DeltaPsim of cells doublely stained with PI and Rh123 were assayed by flow cytometry.

RESULTSThe percentages of sub-G(1) cells after treatment with 1.0 micro g/ml and 2.0 micro g/ml TanIIA had no significant difference but was higher than that of 0.5 micro g/ml. After treatment with TanIIA, NB4 cells appeared the classical apoptotic morphology. The percentages of sub-G(1) cells were increased, while the DeltaPsim reduced (P < 0.01) and there was a linear correlation between them. The increment of sub-G(1) cell percentages and decrement of DeltaPsim induced by TanIIA were partly inhibited by CsA (P < 0.01).

CONCLUSIONSTanIIA can induce NB4 cells apoptosis through opening the mitochondrial permeability transition pore and reducing DeltaPSgr;m, and this effect could be inhibited by CsA.

Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; DNA ; metabolism ; Diterpenes, Abietane ; Humans ; Membrane Potentials ; drug effects ; Mitochondria ; drug effects ; Phenanthrenes ; pharmacology ; Tumor Cells, Cultured

s correlated with coronary disease. Examining these peripheral arteries, more efficient in combination, seems to be a helpful way for screening coronary disease.