Objective To probe into the mechanisms of bone morphogenetic protein-2 (BMP-2) in osteosarcoma and to provide basis for gene therapy. Methods A 1.0 kb cDNA fragment of human BMP-2 gene was inserted reversely into PDOR and Human antisense BMP-2 retrovirus expression vector was constructed. The recombinant retroviral vector was transfected into human osteosarcoma cells OS-9901 with liposome AMINE that expressed abundant BMP. Positive cell clones were selected with G 418. The expression of BMP and proliferating cell nuclear antigen (PCNA) was determined by immunohistochemical ABC methods. The osteosarcoma cell cycle was analysed by flowcytometry, and the changes were observed by electronmicroscope. Results The expression of cellular BMP and PCNA in the transfected osteosarcoma cells decreased obviously (t=24.01， 26.09, respectively, P

objective: To study the effects of BMP on the growth of osteosarcoma cells. Methods: Human antisense BMP2 retrovirus expression vector was constructed and transfected into human osteosarcoma OS 9901 cells by Lipofect AMINE. Positive cell clones were selected with G 418. The expression of BMP and PCNA were determined by immunohistochemical methods, image analysis and expressed as grey level. The morphology and cell cycle distribution of the cells were studied by electronmicroscope and flowcytometry respectively. Results:The grey levels of BMP and PCNA in the transfected cells were 198.4?8.51 and 197.3?3.22, those of the control were 135.1?12.32 and 142.9?8.47,respectively. G 1, G 2 and S phase cells were 51.9%,18.2% and 22.7% in the cell cycle of transfected cells, while those of the control were 52.8%,11.1% and 36.1%, respectively.Increased lysosome and small pieces of chromatin were observed in the transfected cells under electronmicroscope. Conclusion:Transfection of antisense BMP2 may inhibite the expression of BMP and proliferating activity of osteosarcoma cells.

Objective To summarize imaging features of Kallmann syndrome (KS) in children and to improve diagnostic level of the disease.Methods The imaging manifestations of 13 patients with clinically proved KS were retrospectively studied.MRI of rhinencephalon and left wrist X-ray examinations were performed in all the 13 children.Ultrasound of abdomen and pelvis was implemented in the 8 of 13 patients.Results All the patients had abnormalities of olfactory system.Bilateral olfactory bulbs and tracts were absent in 9 patients.Unilateral olfactory bulbs and tracts were absent in 4 patients,in which contralateral olfactory bulbs and tracts were hypoplastic.These patients presented 2 aplastic,17 hypoplastic and 7 normal olfactory sulci.Anterior pituitary was hypoplastic in 3 patients.Bone age showed retardation in 6 of 13 patients.Bilateral testicles were small in all the 8 cases by ultrasound.Conclusions Aplastic or hypoplastic olfactory bulbs and tracts and sulci are seen in children with KS.Anterior pituitary dysplasia is present in part of patients and bone age backward happens in approximate half of the children.

Embryonic stem cells(ESCs) are pluripotent cells derived from the inner cell mass of blastocyst-stage embryos, possessing permanent self-renewal and indefinite proliferative capacity in vitro. And ESCs could differentiate into the hematopoietic cell fate. Therefore ESCs may provide an alternative for hematopoietic stem cell transplantation and blood cells transfusion. Furthermore, ESCs differentiation could also provide a powerful model system to better understand the hematopoietic development and the mechanism involved. The current status for efforts to differentiate ESCs into hematopoietic lineages were reviewed.

Aim To investigate the effects of panax nonstaining saponins ( PNS ) on the apoptosis of renal cells induced by cisplatin through the path of mitochon-drion . Methods Male Sprague-Dawley( SD) rats were randomized divided into normal control group, cisplatin model group and the cisplatin+PNS group,with 12 rats of each group. Animals were sacrificed to determine the N-acetyl-β-D-Glucosaminidase ( NAG ) in urine, blood urea nitrogen ( BUN) and serum creatinine ( Cr) concentrations after 8d of intraperitoneal injection. HE-staining was employed to observe renal pathologies. Transmission electron microscopy ( TEM ) was em-ployed to observe the mitochondria of the rats′ injured kidney region. TUNEL staining was employed to detect the distribution of apoptotic cells. Immunnohistochem-istry was used to detect Bax and caspase-9 expression, and expression of Bcl-2 protein was detected by West-ern blot. Results Compared with the normal control group, contents of BUN, serum Cr and urinary NAG levels of rat in the cisplatin model group were increased ( P <0. 01 ) , and some mitochondria of the epithelial cells of renal tubules got injured seriously. The apopto-sis rate of kidney cell was increased ( P<0. 01 ) . The expression of Bax,caspase-9 and Bcl-2 proteins was in-creased ( P < 0. 01 ) . Compared with the cisplatin model group, contents of BUN, serum Cr and urinary NAG levels of rats in the cisplatin model group were decreased ( P <0. 01 ) , and some mitochondria of the epithelial cells of renal tubules were significantly im-proved. The apoptosis rate of kidney cell was decreased ( P<0. 01 ) . The expression of Bax and caspase-9 pro-tein was decreased (P<0. 01),but Bcl-2 protein was increased ( P < 0. 01 ) . Conclusion PNS may in-crease the expression of Bcl-2 protein and decrease the expression of Bax and caspase-9 proteins, which may play a protective role in cisplatin nephritic damage.

To clone, express and primarily use human autoantigen Sm D1 in methylotrophic yeast Pichia Pastoris. The gene Sm D1 was cloned by PCR.The PCR product was inserted into the vector pPIC9k. The recombinant plasmid pPIC9k- Sm D1 was transformed into yeast SMD1168 by electroporation. The positive clones were screened in MD plates. The high copy number transformants were rapidly selected by using G418 and were induced by methanol. Supernatants after induction were analyzed by SDS-PAGE and im-munodot. The PCR product was showed about 360 bp in size which was in accordance with predicted. The pPIC9k-Sm D1 showed the same seqencing result with GenBank’s report and restriction enzyme analysis confirmed our prediction. The pPIC9k-Sm D1 positive clone produced an about 16 kD protein which had natural immunogenicity of human autoantigen Sm D1 by SDS-PAGE and immunodot. The sensitivity and specificity of immunodot were 96% and 100%, respectively. The agreement between immunodot and im-munoblot was 98%. Successfully cloning and high-level expression of human autoantigen Sm D1 in methy-lotrophic yeast Pichia pastoris laid a foundation for further research work.

0.05 ) when two doctors measured the same group patients with the same method. The coefficient of variation of IVRTd was bigger than that of IVRTm when two doctors measured the same group patients with two different methods. In four sets of data, three ones showed significant difference(P

Objective To obtain recombinant human Smith D1 (Sm D1) antigen and establish detecting assay.Methods Human Smith D1 antigen was synthesized by PCR using human Leukemic cDNA. The prokaryotic expression vector pGEX-ST-Sm D1 was constructed and transformed into E.coli.BL21 cell.Protein expressed under the induction of IPTG.We established DIGFA for detecting anti-Sm D1 antibodies with purified Sm D1 antigens.Results Sequence and restriction analysis revealed Sm D1 gene was cloned in frame into pGEX-5T,SDS-PAGE profile showed a clear protein band with a relative molecular weight of 39 000 and western blotting indicated that the expressed product specifically reacted to polyclonal anti-human Sm D1 genes.There was no significant difference between DIGFA and IB.The agreement between DIGFA and IB was 91.7% as calculated by Kappa statistical method.The sensitivity and specificity of DIGFA were 100% and 83.3% repectively.Conclusions Human Sm D1 gene is successfully cloned、 expressed and purification.The DIGFA,using purified Sm D1 antigens,is as good as IB,rather simpler, more rapid and reliable assay.

Aim To research the protective effect and mechanisms of taurine on myocardial ischemia and reperfusion injury in rats. Methods Myocardial ischemia reperfusion models were established in SD rats. The effects of taurine on the size of myocardial infarction and the changes of activity of SOD , and the levels of MDA and NO in myocardium and plasma of rats with myocardial ischemia and reperfusion injury were observed. Results In rats with myocardial ischemia and reperfusion injury, the taurine could reduce the size of myocardial infarction (P

Objective To investigate the usage of antiepileptic drugs (AEDs),epileptic seizures,pregnancy outcomes and infant feeding practices among pregnant women with epilepsy,and provide scientific evidence for eugenics.Methods The clinical characteristics of 350 pregnant women with epilepsy from Shaanxi Provincial Epilepsy & Pregnancy Register from October 2012 to July 2016 were analyzed retrospectively.Results A total of 350 patients with 376 pregnancies were included in the final analysis.Among 376 pregnancies,272 pregnancies (72.3％) were under the treatment of AEDs.Of them,246 pregnancies (90.4％) were on monotherapy,26 (9.6％) on polytherapy.Only patients during 179 pregnancies (47.6％) took folic acid in the first trimester.Seizure frequency increased in patients during 123 pregnancies (32.7％),of whom patients during 43 pregnancies (35.0％) did not use AEDs.Although other patients during 80 pregnancies (65.0％) took AEDs,regular pharmacokinetic monitoring was absent.A total of 170 pregnancies (45.2％) attained seizure-free.Seizure frequency decreased in 42 pregnancies (11.2％) and remained unchanged in 24 pregnancies (6.4％).The rates of cesarean section and natural delivery were 43.8％ (158/361) and 56.2％ (203/361),respectively.Totally 4.5％ (17/376) pregnancies had adverse pregnancy outcome,and 1.9％ (7/376) had fetal malformations.The most common congenital malformation was heart malformation,which was observed in two offspring.One offspring developed dysostosis,one developed chromosome abnormality,and another developed cerebellum deformity.In addition,one twin developed conjoined deformities.Breastfeeding rate was 52.6％ (190/361).Conclusions In Shaanxi province,AEDs compliance,pharmacokinetic monitoring and usage rate of folic acid still need to be improved in pregnant women with epilepsy.Clinical trial registration Chinese Clinical Trial Registry,ChiCTR-OOC-16009277