The rice gall dwarf disease, caused by the Rice gall dwarf virus (RGDV) is a serious disease occurring in rice in many regions of Guangdong province. As a basis to control the disease we have studied the genomic diversity of a variety of isolates from different locations. Genome segment 8(S8), encoding a main outer capsid protein (Pns8) of RGDV five isolates (BL, CH, DQ, GZ, XY) from Guangdong province was cloned and sequenced. The results revealed that all the S8 segments of the five isolates consisted of 1 578 nucleotides and had a single open reading frame (ORF) extending for 1 301 nucleotides from nucleotide 21 which encoded a polypeptide of 426 amino acids with an estimated molecular weight of 47.4 kDa. The S8 full-length sequence and the ORF sequence shared 97.3%-98.8% and 97.3%-99.1% nucleotide sequence identities within the five Chinese isolates, and shared 94.8%-95.6% and 95.0%-96.0% identities with those of the Thailand isolate respectively. The deduced amino acid sequence of Pns8 in GZ isolate was identical to that in the Thailand isolate, while the amino acid sequence variability of Pns8 within five Chinese isolates ranged from 0.5% to 2.1%. These results indicate that the S8 segment of RGDV is highly conserved in different isolates from different locations. The S8 cDNA from the XY isolate was cloned into the plasmid vector pET-28b(+) and a fused expression protein with an apparent molecular mass of 51kDa was specifically detected in an analysis of Escherichia coli Rossetta(DE3)Ⅱcells. To our knowledge, this is the first report on analysis of the RGDV segment 8 sequence and genetic comparison of different RGDV isolates and their protein expression.

Objective To observe the effect of HeiHuDan（HHD） in the treatment of osteoarthritis（OA） and the proliferation and apoptosis of chondrocytes．Methods The anterior，posterior Cruciate Ligaments and medial menisci of the left knee in 36 adult New Zealand rabbits were sectioned to produce models of OA；the animals were then randomly divided into test-group and control-group．HHD 1 tab per day were given to the animals in the test group for 13 weeks after surgery．16th，20th，and 24th weeks after operation，the animals were sacraficed respectively and specimens were procured from the medial femoral condylar and the medial plateau of tibia for light microscopic，transmission electron microscopic and scanning electron microscopic study．Immunohistochemical method and in situ end labbelling method were used to detect the proliferation and apoptosis of chondrocytes in articular cartilage． Results The degenerative changes of cartilage in test-group had no significant difference from control’s，the apoptosis index and proliferative index of chondrocytes were less than control’s （P＜0.05） at 24 weeks postoperation．Conclusion The study indicated that HHD can decrease the apoptosis of chondrocytes in the late stage of OA and keep the balance of proliferation and apoptosis of OA chondrocytes．

Osteonecrosis of the femoral head (ONFH) is a common refractory disease in orthopedics, and the rate of disability is very high, and its pathogenesis is unknown. TCM has unique advantages in the prevention and treatment of ONFH, and it has gradually received attention. This article reviewed the clinical application status of TCM for prevention and treatment of ONFH in recent years from ONFH's TCM name, etiology, pathogenesis, syndrome differentiation, and clinical treatment, with a purpose to provide a reference for the clinical treatment of ONFH.

BACKGROUND: In recent years, precision medicine has become a hot topic in the medical field. The application of precision medicine to osteonecrosis of the femoral head is wider than before. OBJECTIVE: To summarize the current status and progress of precision medicine to osteonecrosis of the femoral head. METHODS: A computer-based search of PubMed and CNKI databases was performed for literatures about application and prospect of precision medicine for osteonecrosis of the femoral head in and outside China published within the past 10 years using the keywords of "precision medicine, osteonecrosis of the femoral head, three dimensional (3D) printing, computer assisted navigation, minimal invasive, gene" in English and Chinese. RESULTS AND CONCLUSION: (1) In the study on the prevention and treatment of osteonecrosis of the femoral head, 3D printing technology, computer assisted navigation technology, precision minimally invasive orthopedic technology, research and treatment of gene are four effective ways to achieve precise and personalized healthcare. (2) The appearance of precision medicine, a novel medicinal model, enriches the treatment of osteonecrosis of the femoral head and enhances the curative efficacy of the disease. It brings the new dawn to the patient and provides some new clues for medical workers and researchers engaged in the clinical and basic researches of osteonecrosis of the femoral head.

BACKGROUND: Precision medicine pays attention to the depth characteristics of the disease and the high precision of the treatment, and is a precise and accurate personalized approach of the diagnosis and treatment of the disease. In recent years, it has gradually become the hotspot of the medical field. As a new concept of medicine and medical model, its value in clinical treatment of orthopedics has been gradually highlighted. OBJECTIVE: To summarize the current status and progress of precision medicine in the field of orthopedics.METHODS: A computer-based search of PubMed and CNKI databases was performed for literatures about application and prospect of precision medicine in the field of orthopedics in and outside China published within the past 10 years using the keywords of "precision medicine, Orthopedics, bone, 3D printing, computer assisted navigation, minimal invasive, gene". RESULTS AND CONCLUSION: (1) In the study on the prevention and treatment of the disease of orthopedics, 3D printing technology, computer assisted navigation technology, precision minimally invasive orthopedic technology, and gene research are four effective way to achieve precise and personalized healthcare. (2) The appearance of precision medicine, a novel medicinal model, enriches the treatment of the disease in the field of orthopedics and enhances the curative efficacy of the disease. Precision medicine brings the new dawn to the patient and provides some new clues for medical workers and researchers engaged in the clinical and basic research of the disease in the field of orthopedics.

OBJECTIVETo study osteoblastic phenotype expression of New Zealand rabbit skin fibroblasts transfected with mouse core binding factor a1/osteoblast specific transplanting factor-2 gene (Cbfa1/Osf2).

METHODSCbfa1/Osf2 gene, engineered into eukaryotic expression vector pSG5, was introduced into New Zealand rabbit skin fibroblasts with catholyte liposomes-Lipofectamine 2000. Meanwhile, those transfected pSG5 and un-transfected were set the control groups. The expression of Cbfa1 gene, osteocalcin (OCN) gene, alkaline phosphatase (ALP) gene and pre-peptide 2 alpha gene of collagen type I were detected by RT-PCR assay. Cbfa1 protein was detected by Western-Blot assay, in-cell ALP activity by p-nitrophenyl phosphate (PNPP) assay and OCN content in the supernatant by radio-immunity method. The ossification nodules was detected by Alizarin-Red staining and scanning electron microscope.

RESULTSCbfa 1mRNA and Cbfa1 protein were expressed in New Zealand rabbit skin fibroblasts transfected with pSG5-Cbfa1/Osf2 from the first day to the fifth day, but they were not detected in the control groups. In the pSG5-Cbfa1/Osf2 transfected group, the expression of ALP gene and OCN gene were respectively induced from the third day and the forth day, pre-peptide 2 alpha gene of collagen type I was enhanced from the third day. From the sixth day, ALP activity greatly increased, OCN strongly secreted, and they were maintained at a high level for about 4 weeks, and the difference was significant compared with the control group (P < 0.05). On the forty-second day, ossification nodules were found on the surface of pSG5-Cbfa1/Osf2 gene transfected cells.

CONCLUSIONSNew Zealand rabbit skin fibroblasts transfected with pSG5-Cbfa1/Osf2 can express osteogenesis-related genes and proteins, and form ossification nodules on their surface.

Alkaline Phosphatase ; biosynthesis ; genetics ; Animals ; Cell Adhesion Molecules ; biosynthesis ; genetics ; Cells, Cultured ; Core Binding Factor Alpha 1 Subunit ; biosynthesis ; genetics ; Fibroblasts ; cytology ; physiology ; Gene Expression ; Genetic Vectors ; Mice ; Osteocalcin ; biosynthesis ; genetics ; Osteogenesis ; genetics ; physiology ; Rabbits ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection

OBJECTIVETo investigate changes in serum insulin-like growth factor-1 (IGF-1) and insulin-like growth factor-binding protein-3 (IGFBP-3) and their significance in children with left-to-right shunt congenital heart disease (CHD) associated with heart failure (HF).

METHODSTwenty healthy children (control group), 20 children with HF, without basic heart disease (HF group), 20 children with left-to-right shunt CHD, without HF (CHD group), and 30 children with left-to-right shunt CHD associated with HF (CHD+HF group) were included in the study. These groups were compared in terms of serum IGF-1 and IGFBP-3 levels. According to the New York Heart Association (NYHA) Functional Classification, the CHD+HF group was further divided into NYHA-II, NYHA-III and NYHA-IV subgroups and the subgroups were compared in terms of serum IGF-1, IGFBP-3, and cardiac troponin I (cTnI) levels. The correlation of serum IGF-1 and IGFBP-3 levels with serum cTnI level in the CHD+HF group was analyzed.

RESULTSThe CHD group showed decreased serum IGF-1 and IGFBP-3 levels compared with the control group (P<0.01). The CHD+HF group showed a significantly decreased serum IGF-1 level compared with the control group (P<0.01) and CHD group (P<0.05). The HF group had significantly increased serum IGF-1 and IGFBP-3 levels compared with other groups (P<0.01). The NYHA-II subgroup had the highest serum IGF-1 level and the NYHA-IV subgroup had the lowest serum IGF-1 level (P<0.01). In the CHD+HF group, serum IGF-1 and IGFBP-3 levels were negatively correlated with serum cTnI level (r=-0.692, P<0.05; r=-0.530, P<0.05).

CONCLUSIONSSerum IGF-1 level can be used as an objective condition evaluation indicator for CHD, and low serum IGF-1 level is a risk factor for HF. This also provides a clinical basis for treatment of HF using exogenous IGF-1.

Child, Preschool ; Female ; Heart Defects, Congenital ; blood ; Heart Failure ; blood ; Humans ; Infant ; Insulin-Like Growth Factor Binding Protein 3 ; blood ; Insulin-Like Growth Factor I ; analysis ; Male ; Troponin I ; blood

OBJECTIVETo study the feasibility of osteogenic phenotype expression by human skin fibroblasts induced in polyglycolic acid (PGA) foams and the effect of tumor necrosis factor-alpha (TNF-alpha) on the expression of bone morphogenetic protein (BMP) receptors.

METHODSThe fibroblasts were isolated, purified from human skin. (1) Fibroblasts were seeded onto PGA foams. The cell-PGA complexes were cultured in RCCS for 6 weeks, in the media of TNF-alpha (50 U/ml) and BMP-2 (0.1 microg/ml). 1 d, 3 and 6 weeks later, cells and extracellular matrix were investigated by electron microscopic and histochemistry observation respectively. Secretion of osteogenic markers were analyzed by biochemical methods. (2) Fibroblasts were seeded on the glass fragments or culture flasks and treated with TNF-alpha (50 U/ml) in different usage (one-time, all-time). The RT-PCR method and the immunohistochemistry fluorescence staining were used to examine the influence of TNF-alpha on the mRNA expression and the protein expression of the type I BMP receptors at 2, 4, 6, 8 d after treatment.

RESULTSFibroblasts seeded on the PGA foams formed 3-dimensional matrix 3 weeks after seeding, which was demonstrated as osteo-tissue by tetracycline labeling and ARS staining. Cells secreted much more bone-specific alkaline phosphatase (B-AKP) and osteocalcin (OCN) into supernatant than the cells that were cultured in the media without TNF-a and BMP2. Eight days after all-time usage, the TNF-alpha (50 U/ml) increased the expression of the mRNA and protein of the type IB BMP receptor.

CONCLUSIONSFibroblasts on 3-D cell-foam structures can express osteoblastic phenotype under certain inducing conditions. The numerous fibroblasts in body would be a promising resource for cell seeds candidate of tissue- engineered bone. TNF-alpha provides the essential condition for BMP2's target effect on fibroblasts, and combined use of TNF-alpha and BMP2 is one of the regulating factors.

Bone Morphogenetic Protein 2 ; Bone Morphogenetic Protein Receptors, Type I ; biosynthesis ; genetics ; Bone Morphogenetic Protein Receptors, Type II ; biosynthesis ; genetics ; Bone Morphogenetic Proteins ; pharmacology ; Cell Culture Techniques ; methods ; Cell Differentiation ; drug effects ; Cells, Cultured ; Drug Synergism ; Fibroblasts ; cytology ; drug effects ; metabolism ; Humans ; Phenotype ; Polyglycolic Acid ; Transforming Growth Factor beta ; pharmacology ; Tumor Necrosis Factor-alpha ; pharmacology

OBJECTIVETo study the role of serum from asphyxiated neonates in the inducement of human renal proximal tubular epithelial cells (HK-2) adhesion to neutrophils and possible mechanisms.

METHODSHK-2 cells were cultured randomly with 20% serum from neonates (1, 3, and 7 days after asphyxia), pyrrolidine dithiocarbamate (PDTC) or placebo. The activity of myeloperoxidase (MPO), an indicator of adhesion ability of HK-2 cells to neutrophils in suspensions, was detected by the biochemistry assay. Intercellular adhesion molecule-1 (ICAM-1) and nuclear factor-kappaB (NF-kappaB) of HK-2 cells were examined with the immunohistochemical staining.

RESULTSThe expression of MPO in the post-asphyxial 1-day serum treatment group were significantly higher than that in the PDTC treatment and the control groups as well as the post-asphyxial 3 and 7-day serum treatment groups (P<0.01). The expression of ICAM-1 and NF-kappaB in the post-asphyxial 1-day serum treatment group was also significantly higher than that in the other groups (P<0.01).

CONCLUSIONSSerum from asphyxiated neonates can induce HK-2 cell adhesion to neutrophils, possibly through activating NF-kappaB and increasing the synthesis and expression of ICAM-1 on the surface of renal tubular epithelial cells.

Asphyxia Neonatorum ; blood ; complications ; Cell Adhesion ; Cells, Cultured ; Humans ; Infant, Newborn ; Intercellular Adhesion Molecule-1 ; analysis ; biosynthesis ; Kidney Tubules ; pathology ; NF-kappa B ; analysis ; metabolism ; Neutrophils ; physiology

OBJECTIVETo study the osseointegration of the nanophase hydroxyapatite biograde-coated implants and host bone.

METHODSNanophase hydroxyapatite biograde-coated implants, hydroxyapatite biograde-coated implants and noncoated Ti-6Al-4V implants were inserted into both femoral of Beagle canines the tissue response, dynamic osteogensis and SEM were studied at 4, 8 and 12 weeks.

RESULTSThe degradation of nanophase hydroxyapatite was slower than hydroxyapatite, dynamic osteogensis and the form of osteoblast were better than the control groups.

CONCLUSIONThe biological reaction of Nanophase hydroxyapatite biograde-coated implants is better than hydroxyapatite coated implant.

Animals ; Bone Substitutes ; chemistry ; Coated Materials, Biocompatible ; chemistry ; Dogs ; Durapatite ; chemistry ; Male ; Materials Testing ; Nanoparticles ; Osseointegration ; physiology ; Surface Properties