Using the tissue culture technology to achieve the fast propagation of P.ternata and through the photosynthetic characters and yield comparison,two high-yield indivaduals were obtained. The result showed that the coefficent of propagation can get to 1∶6.23 per generation, through 3 month culture the average diameter of the in vitro tuber can get 0.88 cm. P.ternata was a typical shade-plant, of which LCP was only 700?mol?photons/s?m2. Photo-inhibition was observed among all the three leaf-types of P.ternata, and the willow leaf type showed the most obvious photo-inhibition. Both of the light response Pn curve and single tuber weight were significant different among leaf styles. Result for studying the chlorophyll fluorescence parameters showed that all the difference of NPQ, qP and qN among three leaf-types of P.ternata were significant and the difference of potential-photosynthetic productivity among them was significant. The willow leaf style had the strongest light-tolerance capacity.The individuals of T2(peach-leaf type) had the highest Pn and single tuber weight among 11 superior individuals and the individuals of L2(willow-leaf type) showed the strongest light-tolerance capacity. Through 6 month propagation ten thousands in virto tuber of T2 and L2 each were gotten. And after growing in Wenzhou experimental base for one year, the total gross weight of the high-yield individual tuber get to 100kg each.

Objective To evaluate the effectiveness of arthroscope-assisted surgical technique in treating knee fractures and knee peripheral fractures.Methods From September 2000 to December 2005,there were 46 cases with knee or knee peripheral fractures,in whom artbroseope-assisted surgical technique was performed in 36 cases including 24 males and 12 females,with mean age of 30.5 years (18-52 years).Of 36 cases,26 cases with patella fractures were treated by internal fixation of tension band using cancellous bone screws,four with distal end femoral fractures by retrograde interlocking using intramedullary nail and six with fractures of tibial plateau and tibial condyle crista by internal fixation u- sing cancellous bone screws.Results The mean hospitalization was eight days.A follow up for 3-36 months(average 14 months)in 36 cases showed fracture healing.Of 36 cases,32 cases regained full knee range of motion,four left with terminal flexion,with mean loss of 15?,and had thigh circumference (10 cm above knee)1.1 cm(range 0.6-2.0 cm)shorter than health side and two had slight pain in the knee joint.Lysholm knee joint function score was 86.5+7.6 for patellar fractures,tibial plateau frac- tures and condylar crista fractures.Conclusion As for knee or knee peripheral fractures,arthroscope- assisted surgical technique can provide fine monitoring,decrease trauma and complications,fixate inter- nally and help early ground exercise.

Functional analysis of new genes is playing a central role in postgenomic era. Here we reviewed several main strategies including bioinformatics, gene transduction, antisense technology, certain gene silence induced by RNA interference (RNAi), transgene and gene knockout and artificial chromosome transduction.
Animals ; Computational Biology ; methods ; Genes ; physiology ; Humans ; Transduction, Genetic ; methods

The OPG/RANKL/RANK system plays an important role in osteoclastogenesis and represents a great progress in bone biology. RANKL, which expresses on the surface of osteoblast/stromal cells and activated T cells, binds to RANK on the osteoclastic precursors or mature osteoclasts, and promotes osteoclastogenesis and bone resorption. While osteoprotegerin (OPG), which is expressed by osteoblasts/stromal cells, strongly inhibits bone resorption by binding to its ligand RANKL and thereby blocks the interaction between BANKL and RANK. A number of cytokines and hormones exert their effects on bone metabolism by regulating the OPG/RANKL ratio in the bone marrow microenvironment. RANK is also expressed on mammary epithelial cells and RANKL expression in these cells is induced by pregnancy hormones, RANKL and RANK are essential for the formation of the lactating mammary gland and the transmission of maternal calcium to neonates in mammalian species. Modulation of these systems provides a unique opportunity to develop novel therapeutics to inhibit bone loss in osteoporosis, rheumatoid arthritis, and bone metastasis of cancer. Further research should be focused on the cooperation of OPG/RANKL/RANK system with other signal pathways and the interactions among bone remodeling, immune system and endocrinology system. Currently, the development of OPG analogues or compounds which may stimulate OPG expression is becoming an attractive industry which may be profitable to both patients and manufacturers.
Animals ; Bone Resorption ; immunology ; metabolism ; Humans ; Osteoclasts ; cytology ; metabolism ; pathology ; Osteogenesis ; drug effects ; genetics ; immunology ; Osteoprotegerin ; metabolism ; physiology ; RANK Ligand ; metabolism ; physiology ; Receptor Activator of Nuclear Factor-kappa B ; metabolism ; pharmacology ; physiology ; T-Lymphocytes ; drug effects ; immunology

OBJECTIVETo express human osteoprotegerin (OPG) in E. Coli and analyze its bioactivity in vitro.

METHODSSynthetic oligonucleotides were used to amplify human OPG gene by RT-PCR from total RNA of human osteosarcoma cell line MG63. The OPG cDNA coding for 380 amino acid residues was inserted into prokaryotic expression vector pRSET-A, transformed into competent E. Coli BL21, and induced by 0.1 mmol/l IPTG. SDS-PAGE and Western blot were performed to identify OPG-6His fusion protein. After purified by affinity chromatography, 1,000 microg/L or 1,500 microg/L of OPG-6His were added into the mouse bone marrow cells culture medium. The number of tartrate-resistant acid phophatase (TRAP)-positive multinucleated cells and resorption pits were counted to assess the bioactivity of expression products.

RESULTSThe sequence of OPG mature peptide encoding cDNA obtained in this experiment was as same as reported. SDS-PAGE showed 24% of total bacterial protein was of OPG-6His fusion protein. Western blot assay demonstrated that the molecular weight of recombinant protein was about 46 KD and could react specifically with human anti-OPG antibody. The mouse bone marrow cells were induced by 1alpha, 25-dihydroxyvitaminD3 (10(-8) mol/L) and Dexamethasone (10(-7) mol/L) to form osteoclastic-like multinucleated cells. 1,500 microg/L of purified OPG-6His protein could decrease the number of resorption pits and TRAP-positive multinucleated cells in vitro (P < 0.05), but it didn't show the same effects when the concentration of OPG-6His fusion protein was of 1,000 microg/L.

CONCLUSIONSHuman OPG-6His fusion protein is expressed and purified in E. Coli. The expression products have moderate inhibitory effects on osteoclast differentiation and bone resorption in vitro only when excessive amount of proteins are added into the culture medium, indicating that prokaryotic expression of fuctionalal OPG protein awaits further investigation.

Cell Differentiation ; drug effects ; Cell Line, Tumor ; Cloning, Molecular ; Escherichia coli ; genetics ; Glycoproteins ; biosynthesis ; genetics ; Humans ; Osteoclasts ; drug effects ; physiology ; Osteoprotegerin ; Receptors, Cytoplasmic and Nuclear ; biosynthesis ; genetics ; Receptors, Tumor Necrosis Factor ; Recombinant Fusion Proteins ; biosynthesis ; pharmacology

Using the isolated total RNA from osteosacoma cell line MG63, the cDNA encoding human OPG was amplified by RT-PCR. A recombinant adenoviral vector carrying cDNA of OPG was constructed and OPG expression in mouse myoblast C2C12 cells was confirmed by Western blot and ELISA. The secreted expression of OPG protein persisted more than 6 weeks in vitro, and the growth of C2C12 cells infected by recombinant adenoviral were in good state. Osteoclasts derived from mouse bone marrow cells infected with recombinant adenoviral made less number of TRAP positive cells and resorption pits formed on dentine slices.
Adenoviridae ; genetics ; Animals ; Blotting, Western ; Cell Line ; Enzyme-Linked Immunosorbent Assay ; Genetic Vectors ; genetics ; Humans ; Male ; Mice ; Mice, Inbred BALB C ; Osteoclasts ; metabolism ; Osteoprotegerin ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVEThe traditional gracilis musculocutaneous flap is supplied by a branch of deep femoral artery, which enters the muscle in between the upper and middle third of it. So the flap barely reaches the pelvis and perineum region for reconstruction. By exploring the blood supply pattern we tried to rotate the flap Upon at the higher point starting at the obturator foramen in order to let it cover a bigger area.

METHODSanatomical reviewing of the blood supply of the gracilis branches of obturator, medial femoral circumflex and deep femoral arteries. Based on this a new type of longitudinal gracilis musculocutaneous flap supported only by the obturator artery was designed to reach the pelvis, female genitalia, pubic symphysis, inguinal area easily.

RESULTSThe new kind of flap has been applied to 9 patients for deformity repairing and tissue replacement in the pelvic and perineal area. All the flaps survived and achieved satisfactory result with 3 months to 3 years' follow up.

CONCLUSIONSLongitudinal gracilis musculocutaneous flaps supplied by the obturator artery can be used as regular musculocutaneous flap clinically.

Female ; Femoral Artery ; surgery ; Humans ; Muscle, Skeletal ; blood supply ; transplantation ; Surgical Flaps ; blood supply

OBJECTIVETo observe and compare perioperative myocardial enzyme changes in 107 patients with congenital (CHD, n = 53), rheumatic (RHD, n = 40) and coronary artery (CAD, n = 14) diseases, and to find whether different diseases can affect the release and recovery of myocardial enzymes after heart operations.

METHODSOn the day before operation and the 1st, 3rd, 5th and the 8th day after operation, the venous blood was taken to measure the release of myocardial enzymes: aspartate aminotransferase (AST), creatine kinase (CK), MB isoenzyme of creatine kinase (CK-MB), lactate dehydrogenase (LDH) and LDH-1.

RESULTSAll the enzymes measured before operation in three groups were in the normal range; their release increased abruptly on the 1st day postoperatively to 2 - 15 times of those before operation; on the 3rd day, they recovered to some degrees, and on the 8th day they recovered to normal in all groups except LDH and LDH-1 in rh and CAD groups. Because the aortic cross-clamp time (CCT) had a good positive correlation to the release of myocardial enzymes, those patients whose CCT was over 60 minutes in three groups were compared revealing that the CCT was not different between three groups (P < 0.05). The release of CK, CK-MB and AST was significantly higher in CHD60 group than those in CHD60 and CAD60 groups, they recovered afterwards; while the release of DH and LDH-1 was higher in CAD60 group than those in CAD60 and in CHD60 groups from the 1st day to the 8th day postoperatively.

CONCLUSIONSThe release of all the 5 enzymes measured before operation was in normal range in selected CHD, RHD and CAD patients. The release peak and the recovery order of all enzymes were the same in three groups. The release of CK, CK-MB and AST was higher in CHD60 group than those in RHD60 and CAD60 groups on the 1st day. The release of LDH and LDH-1 was higher in RHD60 group than those in CHD60 and CAD60 groups from the 1st day to the 8th day postoperatively. The shorter the CCT is, the less the release of myocardial enzymes. Using the release of LDH and LDH-1 to evaluate the recovery of myocardial injury after open-heart operations was recommended.

Adolescent ; Adult ; Aspartate Aminotransferases ; blood ; Child ; Coronary Artery Bypass ; Coronary Artery Disease ; blood ; enzymology ; surgery ; Creatine Kinase ; blood ; Creatine Kinase, MB Form ; blood ; Female ; Heart Defects, Congenital ; blood ; enzymology ; surgery ; Humans ; Intraoperative Period ; Isoenzymes ; blood ; L-Lactate Dehydrogenase ; blood ; Male ; Middle Aged ; Myocardium ; enzymology ; pathology ; Rheumatic Heart Disease ; blood ; enzymology ; surgery ; Time Factors

OBJECTIVETo investigate the incidence rate of anterior knee pain after total knee arthroplasty (TKA) and identify the related factors.

METHODSThis prospective, double-blind clinical trial involved a total of 128 patients scheduled for primary ipsilateral cemented three-component TKA for osteoarthrosis. The patients were randomized into two groups to receive operations for TKA with patellar resurfacing (experimental group) or not (control). The incidence of anterior knee pain was observed in these patients and the factor affecting the occurrence of anterior knee pain and knee was analyzed.

RESULTSThe patients were followed up for a mean of 16.5 months (range 6~24 months). The incidence rate of anterior knee pain was 10.9% (7/64) in the experimental group, showing no significant difference from the rate of 14.1% (9/64) in the control group. But the 52 patients with varus or valgus knee showed a significantly higher incidence rate of anterior knee pain (21.2%, 11/52).

CONCLUSIONTKA with patellar resurfacing can not decrease the incidence of anterior knee pain, and varus or valgus before the operation is associated with a higher risk of anterior knee pain.

Aged ; Arthroplasty, Replacement, Knee ; adverse effects ; methods ; China ; epidemiology ; Denervation ; methods ; Double-Blind Method ; Female ; Humans ; Knee Joint ; blood supply ; Male ; Middle Aged ; Osteoarthritis, Knee ; surgery ; Patella ; surgery ; Patellofemoral Pain Syndrome ; epidemiology ; etiology ; physiopathology ; Risk Factors