New studies have shown that metabolism of bone is controlled by the central nervous system.Gonadotropins secreted by the anterior pituitary stimulate the formation and function of osteoclasts which are closely related to the bone turnover and changes of bone mass. This study overviews the relationship between gonadotropins and bone metabolism.

The osteocyte has been found to be an orchestrator of bone remodeling.The damage of bone leads to osteocyte apoptosis.Sclerostin secreted by osteocyte causes feedback inhibition of bone formation,so inhibition of sclerostin expression has become a new target of treatment for osteoporosis.It seems resonable to direct clinical practise and treatment of metabolic bone diseases through understanding the function of osteocyte in the process of bone remodeling.

Objective To isolate and identify advanced oxidation protein products from human serum albumin (AOPP-HSA), expecting to search for a method of preparing highly purified and bioactive AOPP-HSA. Methods AOPP-HSA crude products were prepared in vitro by exposing HSA to HOCl. AOPP-HSA was isolated by gel chromatography and ion-exchange HPLC. Its structural features and biological activities were characterized by UV and fluorescence spectrum, SDS-PAGE, and the experiment of TNF-? release from monocytes. Results The isolated protein was purified up to 99.4% and was dityrosine-containing protein cross-linking products with molecular weight of 700?10 3. It possessed the ability of triggering the considerable release of TNF-? from monocytes. Conclusion Highly purified and bioactive AOPP-HSA can be successfully prepared by above-mentioned two-step chromatography from AOPP-HSA crude products, which builds a basis for further study of AOPP.

Objective To observe the cytotoxicity of Ad-HO-1 and its capacity of mediating the expression of HO-1 in cultured HK-2 cells. Methods The HK-2 cells were infected by Ad-HO-1 with 100, 200 and 400 MOI for 3 h, followed by the green fluorescence observation 2 days later. The live cells ratio was detected 2 and 4 days later. The expression of HO-1 and GFP in HK-2 were detected under laser scan confocal microscope. The expression of HO-1 was detectee by Western blot analysis. 3H-TdR was used to assay the ability of HO-2 cells infected with Ad-HO-1 to inhibit the PBMC proliferation. Results Over 90% HK-2 cells expressed green fluorescence after infected by Ad-HO-1 (MOI 200) for 2 days. The live cell ratio at 2 and 4 days had not any difference from those of the control group. The expressions of HO-1 and GFP in the Ad-HO-1 transfected HK-2 cells were determined under laser scan confocal microscope. The locations of these two proteins were the same. HK-2 cells infected by Ad-HO-1 had protein immune blot strap combined with HO-1 antibody. When the MOI of Adv-HO-1 was 0.01, 0.1, 1 and 10, the proliferative capacity of PBMC was inhibited significantly. Conclusion The stable expression of HO-1 mediated by Ad-HO-1 in cultured tubular epithelial cells can inhibit the cell proliferation of PBMC.

Objective To investigate the effects of HO-1 on renal allografts after HO-1 gene transfection in rat chronic allograft nephropathy model. Methods Twenty F344 and twenty-six Lewis rats were included in this experiment. They were divided into 3 groups at random. Six Lewis rats were in pseudo-operation group, 10 Lewis were in empty carrier group (transfected with adenovirus) and 10 Lewis were in the gene transfection group (transfected with Ad-HO-1 adenovirus). The expression of HO-1 protein was detected by WB at the 1st ,2nd,3rd and 4th week. The content of creatinine in blood was assayed at the 4th,8th, 12th and the 16th week. The weight of rat, the value of patholiogical changes and the expression of ?-SMA,TGF-?1 and PDGF-B were analysized at the 16th week. Results The weight of rats in 3 groups had not any changes at 16th week. The content of creatinine in blood of gene transfection group were lower than those in the empty carrier group. The expression of HO-1 protein were very high in the 1st and 2nd week and decreased at 3rd and 4th week. The Banff value of kidney in the gene transfer group was better than that of the empty carrier group at the 16th week. The level of ?-SMA, TGF-?1 and PDGF-B in the gene transfection group were significantly lower than those in the empty carrier group. Conclusions Ad-HO-1 could efficiently transfer HO-1 gene into rat donor kidney. The prevention of chronic allograft nephropathy may have relationship with the decreasing expression of ?-SMA, TGF-?and PDGF-B.

Objective To explore the effect of ultra-early hyperbaric oxygenation (HBO) on bone calcium, biomechanical properties and bone collagen of femur in rats with complete spinal cord transaction. Methods A total of 75 Sprague-Dawley rats were randomly divided in-to sham group (n=15), model group (n=20) and HBO group (n=40). HBO group was divided into three hours group (HBO1 group, n=20) and twelve hours group (HBO2 group, n=20). All groups underwent laminectomy at T10, while the model group, HBO1 group and HBO2 group underwent complete spinal cord transection at the same level. Three hours and twelve hours after surgery, HBO1 group and HBO2 group received HBO, respectively, for three courses with ten days in a course. After treatment, the femoral biomechanical properties, bone calcium and hydroxyproline (Hyp) were determined. The morphology of bone trabecula and the bone collagen was observed with HE stain-ing and Masson triad color staining, respectively. Results After treatment, compared with the sham group, the femoral biomechanical proper-ties, the content of bone calcium and Hyp decreased in the model group (P<0.05);compared with the model group and HBO2 group, they in-creased in HBO1 group (P<0.05). The number of bone trabecula and the bone collagen decreased, and derangement and sparseness were ob-served in the model group;however, the changes were substantially mild in HBO1 group. Conclusion Ultra-early HBO could increase the content of bone calcium and Hyp of femur, improve the morphology of the femur bone collagen, and improve the femoral biomechanical properties in rats with complete spinal cord transection.

Objective To analyze the genotype-phenotype correlations of Angelman syndrome ( AS ) , and to discuss the advantage of applying array-based single nucleotide polymorphisms comparative genomic hybridization ( SNP aCGH) in diagnosis of AS. Methods Examination of electroencephalogram( EEG) and intelligence quotient( IQ) evaluation were done for 11 cases diagnosed as AS clinically. Gesell scares were chosen as the evaluation criterion of IQ. The screening techniques was methylation polymerase chain reaction( MS-PCR) ,then SNP aCGH was used to make genetic diagnosis. Results (1)Eleven cases of AS were confirmed:1 case had UPD(uniparental disomy),10 cases were type of deletion, from which 6 cases were deletion (Ⅱ) , 4 cases were deletion (Ⅰ) . ( 2 ) The copy number variations were detected in the region of 15q11-q13,which contained genes like MKRN3,MAGEL2,NDN,SNRPN, SNURF,GABRB3,GABRA5,GABRG3,UBE3A,OCA2,ATP10A. To search online Mendelian inheritance in man,genes above were correlated with AS manifestation. (3)All cases of deletion were 3-5 standard deviation(SD) in weight and height to normal children at the same age and with the same sex,while UPD was below 1. 5 SD. Gesell scares showed that the deletion(Ⅰ) was the most serious in mental retardation,deletion(Ⅱ) was moderate,and the UPD was mild. Eight cases were hypopigmentation,and one was the UPD. EEG revealed that 1 case of deletion(Ⅰ) and the UPD were spike occasionally,another one deletion(Ⅰ) was limit EEG. The rest cases displayed slow and spike waves paroxysmal-ly,with amplitude of medium or high,2. 5-3. 0 Hz. Conclusions Not only can SNP aCGH make a diagnosis of AS but discriminate the types of genetic pathology. Since different type contributes to a diverse of clinical features and the rate of recurrence is also different,it is significant for family genetic consultation. Moreover,the technology is advantageous for the study on the pathogenesis and gene function.

Objective To construct HSP27 eukaryotic expression plasmids. Methods Full-length HSP27 gene was amplified by reverse transcription polymerase chain reaction (RT-PCR) from breast cancer cell line MCS-7 and cloned into eukaryotic expression vector pAAV-MCS. After the recombinant plasmids transfected into NIH3T3 cells, the expression of HSP27 protein in the host cells was characterized by ECL Western blotting. Results Full-length HSP27 gene was amplified by RT-PCR correctly. The correct cloning of HSP27 gene in pAAV-MCS was confirmed by restriction enzyme digestion and sequencing. ECL Western blotting results indicated that the target gene could express in the mammalian cell line NIH3T3. Conclusion Recombinant plasmid HSP27/pAAV-MCS had been cloned successfully, which would provide the foundation for investigating the role and the mechanism of HSP27 in the ischemia precondition of kidney.

Objective To explore the effects of aging on NO cGMP pathway and sexual hormone in penile tissue of different age rats. Methods The nitric oxide(NO) and cyclic guanosine monophosphate(cGMP) contents, nitric oxide synthase(NOS) activity in penile tissue and testosterone(T) and luteinizing hormone(LH) contents in blood of rats of different ages(2, 8, 16 and 24 months) were detected. Results With age increasing, ① The NO content of penile tissue firstly rose(8 months, the highest ) then dropped (24 months, the lowest), being the same as the variation of NOS( P

Objective By using array-based single nucleotide polymorphisms comparative genomic hybridization (SNP-aCGH) to detect and fine mapping copy number variations (CNVs) in children with unexplained mental retardation/brain development delay(MR/BD),then the CNVs were analyzed to determine diagnosis and offer genetic counseling,finally to discuss the application of SNP-aCGH in genetic diagnosis of MR/BD with unknown causes.Methods Ninety-two children with unexplained MR/BD were recruited.SNP-aCGH was used to get CNVs from the whole genome-wide,and the correlation of CNVs and phenotype was analyzed to definit disease genes or pathogenic fragment.Statistics was performed to analyze the common phenotype between positive cases (case with CNVs) and negative cases.Results (1) The CNVs were detected in 10 cases with a detection rate of 10.86％,from which 8 cases showed subtelomeric aberration,5 cases without subtelomeric aberration,and the rate was 8.70％,5.40％,respectively.The CNVs related to MR/BD involved 10 different subtelomeric regions (9p,21q,3p,2p,15q,4p,12p,22q,16p,17p),and 7 different regions without subtelomeric (1p,4q,2p,14q,15q,12q,22q).The deletions involved 11 zones (size:1.05-8.80 Mb),and duplications referred to 8 zones (size:1.33-31.25 Mb).(2) One case was diagnosed as 9p duplication syndrome,for candidate genes:DOCK8,VLDLR.A case was detected with a gene fracture (CRBN).One case was diagnosed as Coffin-Sirrs syndrome combined with a deletion of 15q26.3-qter,for candidate genes:SOX11 and LINS1,respectively.One case referred to 12p13.3 deletion syndrome,for candidate genes:ELKS,ERC1.One case referred to 22q13.2-qter deletion,for candidate genes:SHANK3.Two cases were diagnosed as ATR-16 syndrome with 17p13.3 deletion syndrome,their candidate genes:HBA1,HBA2,SOX8 for the former,YWHAE,LIS1 for the latter.(3)There were statistically significant differences in comparison of positive cases to the negative ones for growth delay,internal organs deformity,low birth weight infant(LBW) and premature infant (all P ＜0.05).Conclusions (1) Besides MR/BD in different degrees in all the positive cases,they also showed growth delay,a portion of them with internal organs deformity,low birth weight infant and premature infant.(2) Subtelomeric aberrations are related to MR/BD,while the submicroscopic rearrangement in regions without subtelomeric is suspiciously pathogenic,and need to be further studied.(3) SNP-aCGH can fine mapping the region of CNVs by high resolution from the whole genome-wide,which does contribute to limit the zones for finding pathogenic region and candidate genes,as well as to offer a technology platform for investigating about the correlation of phenotype and genes or CNVs.