Actins ; metabolism ; Adult ; Amyloidosis ; pathology ; Diagnosis, Differential ; Female ; Humans ; Intestinal Pseudo-Obstruction ; metabolism ; pathology ; Lupus Erythematosus, Systemic ; pathology ; Muscular Atrophy ; pathology ; Scleroderma, Systemic ; pathology ; Young Adult

AIM:To investigate the activity of NF-?B in peripheral blood mononuclear cells (PBMCs) from patients with Graves disease (GD) and the significance in immunopathogenesis of GD.METHODS:Peripheral blood was collected from 22 untreated GD,20 treated GD with tapazole more than 1 year,and 25 healthy volunteers. PBMCs were isolated from the blood by histopaque-1077 density-gradient centrifugation. The activity of NF-?B in PBMCs was analyzed using gel electrophoretic mobility shift assay (EMSA). The contents of IL-1?,IL-6 and TNF-? were tested by radioimmunoassay.RESULTS:The activity of NF-?B in PBMCs of untreated GD group was increased remarkably,compared with that in the treated group and control (P

OBJECTIVETo observe dynamically the development of fetal long bone and detect the expression and distribution of HIF-1alpha,to investigate the expression pattern and possible effects of hypoxia inducible factor-1alpha (HIF-1alpha) in fetal long bone development of mouse.

METHODSE12.5, E13.5, E14.5, E15.5, E16.5 and E17.5 pregnant C57BL6 mice were sacrificed. After sacrifice, the embryos were delivered by caesarean section. The development of fetal long bone was dynamically observed by stereoscopic microscope, and the distributional expression of HIF-1alpha protein was detected by using method of immunohistochemistry. The expression of HIF-1alpha mRNA and osteoblast marker gene at various stage were also detected by using methods of reverse transcription-polymerase chain reaction (RT-PCR).

RESULTSThe cartilaginous long bone began to form and joints outline arised at E13.5, then the primary ossification center was observed at E14.5, showing opaque ossification under stereoscopic microscope,and then the osteogenesis expanded and extended to both sides. Immunohistochemistry demonstrated lots of HIF-1alpha protein positive chondrcytes in the center of primary ossification at E14.5, then they decreased dramatically. HIF-1alpha mRNA expressed at high level from E13.5 to E15.5, and then decreased to low level.

CONCLUSIONFetal long bone development pattern appeared to be endochondral osteogenisis process, existing hypoxia microenviroment may increase HIF-1alpha mRNA expression and thus initiate the cascade of endochondral osteogenisis.

Animals ; Bone Development ; Female ; Hypoxia-Inducible Factor 1, alpha Subunit ; analysis ; genetics ; physiology ; Immunohistochemistry ; Male ; Mice ; RNA, Messenger ; analysis

DNA microarrays have been acknowledged to represent a promising approach for the detection of viral pathogens. However, the probes designed for current arrays could cover only part of the given viral variants, that could result in false-negative or ambiguous data. If all the variants are to be covered, the requirement for more probes would render much higher spot density and thus higher cost of the arrays. Here we have developed a new strategy for oligonucleotide probe design. Using type I human immunodeficiency virus (HIV-1) tat gene as an example, we designed the array probes and validated the optimized parameters in silico. Results show that the oligo number is significantly reduced comparing with the existing methods, while specificity and hybridization efficiency remain intact. The adoption of this method in reducing the oligo numbers could increase the detection capacity for DNA microarrays, and would significantly lower the manufacturing cost for making array chips.

BACKGROUNDFibroblast growth factor 9 (FGF9), expressed in brain, kidney and developing skeletal tissues, can physiologically inhibit endochondral ossification; but little is known about how FGF9 affects osteoblasts and its detailed regulatory mechanism. Here we examined the effect of FGF9 on the activity of the murine Runt-related transcription factor 2 (Runx2) gene promoter in preosteoblast MC3T3-E1 and premyoblast C2C12 cells.

METHODSPlasmids containing the Runx2 promoter region were transfected into MC3T3-E1 and C2C12 cells and stably transfected cell lines were established. The method of luciferase reporter gene activation was used to examine the effects of FGF9 on the promoter activity.

RESULTSFGF9 (10 ng/ml) increased Runx2 promoter activity in MC3T3-E1 cells. When MC3T3-E1 cells were treated with FGF9 plus the various inhibitors or activator of the intracellular signaling transducation pathways, including 10 micromol/L U0126 (the inhibitor of mitogen-activated protein kinase kinase), 10 micromol/L SB203580 (the inhibitor of p38/mitogen activated protein kinase), or 1 micromol/L C6 ceramide (an activator of mitogen activated protein kinase), the luciferase expression did not change significantly compared with that of the cells treated with FGF9 only. However, when C2C12 cells were treated with 10 ng/ml FGF9, Runx2 gene promoter activity first decreased and then increased over a period of 1 to 5 days. Among the above inhibitors, only U0126 (10 micromol/L) completely blocked the effects of FGF9 on Runx2 gene promoter activity.

CONCLUSIONSOur data showed that FGF9 can affect Runx2 gene promoter activity in MC3T3-E1 and C2C12 cells. The action of FGF9 appears to depend partly on the mitogen-activated protein kinase kinase/mitogen-activated protein kinase pathways in C2C12 cells.

Animals ; Cells, Cultured ; Core Binding Factor Alpha 1 Subunit ; genetics ; Fibroblast Growth Factor 9 ; pharmacology ; MAP Kinase Signaling System ; Mice ; Myoblasts ; drug effects ; metabolism ; Osteoblasts ; drug effects ; metabolism ; Promoter Regions, Genetic

OBJECTIVETo investigate the effect of fresh meniscal allografts combined with osteochondral allografts transplantation for treatment of osteoarthritis.

METHODSThirty-six rabbits were used in the experiment and were randomly divided into 3 groups: in group A, the fresh medial meniscal allografts combined with osteochondral allografts from medial tibial plateau were implanted into medial articular meniscal and medial tibial plateau osteochondral defects; in group B,the fresh medial meniscal allografts were implanted into medial meniscal allografts defects; in group C, the freezing medial meniscal allografts were implanted into medial meniscal allografts defects. General observation, histology examination and glycosaminoglycan (GAG) examination in cartilage of medial tibial plateau were performed at the 4th, 8th and 12th week after operation.

RESULTSThere were no significant differences in cellular counting and amount of GAG between group A and group B, but the cellular amount of group A was significantly more than that of group C at the 12th week.

CONCLUSIONFresh meniscal allografts combined with osteochondral allografts transplantation can repair meniscal and osteochondral defects.

Animals ; Cartilage ; transplantation ; Female ; Male ; Menisci, Tibial ; transplantation ; Rabbits ; Transplantation, Homologous

[Objective] To improve the understanding of CT and MRI features of giant cell reparative granuloma of temporal bone and reduce misdiagnosis.[Methods] The CT and MR images of 4 cases of GCRG of temporal bone were analyzed,compared with their operation and pathology results.[Results] All the lesions of the 4 cases were located in the anterior and lower parts of the temporal bone with widely destruction.The CT images showed expansive destruction of bone with disruption of osseous shell,strip and punctate calcification and ossification in and around the lesion,and osteosclerosis of the adjacent bone,which consistent with the scope of the operation.The MR images showed a large patchy of low signal intensity on both T1-weighted and T2-weighted images,which showed heterogeneous enhancement after injection of contrast.Fibrous proliferation with multiple multinuclear giant cells and hemosiderin deposition were showed under microscope.[Conclusion] The morphological and pathological characteristics of recurrent intraosseous hemorrhage and parcels of granulation tissue in GCRG of the temporal bone could be reflected by CT and MR images,which has certain characteristics and is of important significance to the diagnosis of the tumor.

OBJECTIVETo investigate the impact of rheumatic heart disease on pregnancy.

METHODSA retrospective study was conducted in 125 pregnant women with rheumatic heart disease. These cases were divided into operation (group A) and non-operation (group B), and the cardiac function, pregnancy complications, gestational weeks, delivery modes and outcomes of pregnancy were compared.

RESULTSThe cardiac function was significantly improved and pregnancy complications reduced after the cardiac operation with extended gestational weeks, showing significant differences between the two groups (P<0.05). One perinatal fetal death without maternal death occurred in group A, as compared with two maternal and two perinatal deaths in group B. Cesarean section was the primary delivery mode in these cases.

CONCLUSIONSurgical interventions can improve the cardiac function, reduce the pregnancy complications and improve the pregnancy outcomes in pregnant women with rheumatic heart disease. Cesarean section is the primary choice for pregnant women with prosthetic heart valve replacement.

Adult ; Cesarean Section ; Female ; Heart Valve Prosthesis Implantation ; Humans ; Pregnancy ; Pregnancy Complications, Cardiovascular ; physiopathology ; surgery ; Pregnancy Outcome ; Retrospective Studies ; Rheumatic Heart Disease ; physiopathology ; surgery ; Young Adult

To compare the DAAO expression level in different Pichia pastoris host strains, the gene encoding DAAO from Trigonopsis variabilis was cloned into plasmid pPIC3.5k and then transformed into P. pastoris GS115 and KM71 respectively. The positive transformants PDK13 (MutS) and PD27 (Mut+) were obtained by PCR analysis. Their optimal and different expression conditions were investigated. To compare with PD27, PDK13 was determined to poss a slower consumption of methanol, a longer induction time, a lower oxygen request and apparently higher expression of DAAO. The highest expression levels were reached up to 2700, 2500 IU/L in shaking flask and 10140, 8463.5 IU/L in fermentor respectively. The over-expression of DAAO can meet its large demand for production of 7-ACA, alpha-keto acid and L-amino acid. In addition, the phenylpyruvate and L-phenylalanine were obtained by crude DAAO reacting with DL-phenylalanine at 37 degrees C for 3h.
D-Amino-Acid Oxidase ; genetics ; Fermentation ; Methanol ; metabolism ; Phenylalanine ; metabolism ; Pichia ; genetics ; Polymerase Chain Reaction

Using of two-step integrating technology, transducted the H and L chain gene of humanized Fab fragment of anti-HB-sAg antibody into the genome of methylotropic yeast P. pastoris. Constructed a engineering yeast to produce humanized Fab fragment of the anti-HBsAg antibody. The Fab fragment was efficiently secreted into the medium at a concentration of 50-80 mg/L. The Fab fragment was purified from culturing supernatant of the recombinant yeas by affinity chromatography. The ELISA analysis showed the high affinity of the expressed humanized Fab fragment to the HBsAg.
Chromatography, Affinity ; Cloning, Molecular ; Enzyme-Linked Immunosorbent Assay ; Hepatitis B Antibodies ; biosynthesis ; genetics ; isolation & purification ; Hepatitis B Surface Antigens ; immunology ; Humans ; Immunoglobulin Fab Fragments ; biosynthesis ; genetics ; isolation & purification ; Pichia ; genetics ; Recombinant Proteins ; biosynthesis ; isolation & purification