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 investigate the role of vascular endothelial growth factor (VEGF) in the pathogenesis of severe acute pancreatitis (SAP) in rats.

METHODSSixty-four male SD rats were randomly divided into control group and SAP group, and in the latter group, SAP was induced by retrograde injection of 5% sodium taurocholate in the pancreaticobiliary duct. The rats were sacrificed at 1, 3, 6 and 12 h after the operation, and the severity of pancreatitis was assessed according to histological scoring. The serum levels of VEGF were examined with enzyme-linked immunosorbent assay, and the expression of VEGF in the pancreatic tissues was measured by SP immunohistochemistry. Another 30 SD rats were randomized into the control group, SAP group and SAP+recombinant rat VEGF injection group, and the vascular permeability of the pancreatic microcirculation was determined by Evans Blue leakage test.

RESULTSAt each of the time points for measurement, both the serum VEGF level and scores of pancreatic tissue injury were significantly higher in SAP group than in the control group (P<0.05). Compared with the control group, the expressions of VEGF in the pancreatic tissues of SAP group were significantly up-regulated following the operation (P<0.05). The vascular permeability of the pancreatic microcirculation significantly increased after the onset of SAP, and injection of recombinant rat VEGF significantly increased the leakage rate of Evans Blue.

CONCLUSIONVEGF may play an important role in the pathogenesis of pancreatitis and in causing edema and hemorrhage in SAP, and the level of serum VEGF may reflect the severity of pancreatic injury.

Acute Disease ; Animals ; Biomarkers ; Capillary Permeability ; physiology ; Male ; Pancreatitis ; metabolism ; pathology ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Vascular Endothelial Growth Factor A ; blood

OBJECTIVETo investigate the protective effects of captopril against lung injury in a rat model of severe acute pancreatitis (SAP).

METHODSSeventy-two male SD rats were randomized into sham-operated group (SO group), SAP group and captopril intervention group (CAP group). Serum amylase and myeloperoxidase (MPO) activity in the lung tissue were examined at 1, 6 and 12 h after the operation. TNF-α and AngII in the lung tissue were detected by ELISA, and the histopathological changes of the pancreas and lung were observed microscopically.

RESULTSThe MPO activity , which was similar between SAP group and CAP group at 1 h, were significantly lowered in CAP group at 6 and 12 h (P<0.05). Serum amylase level and the levels of TNF-α and AngII in the lung tissue homogenate were all reduced significantly in CAP group as compared to those in SAP group (P<0.01). The pathological injury of the lung was obviously lessened in CAP group in comparison with that in SAP group.

CONCLUSIONCaptopril can ameliorate SAP-induced lung injury in rats.

Amylases ; blood ; Angiotensin II ; metabolism ; Animals ; Captopril ; pharmacology ; therapeutic use ; Disease Models, Animal ; Lung ; metabolism ; pathology ; Lung Injury ; etiology ; prevention & control ; Male ; Pancreatitis ; complications ; drug therapy ; Peroxidase ; metabolism ; Rats ; Rats, Sprague-Dawley ; Tumor Necrosis Factor-alpha ; metabolism

Objective: To investigate the changes of myo cardial contractile function during myocardial stunning in calcium overload rats and the protective effects of tetrandrine. Methods: Forty-six rats were randomized into control, myocardial ischemia, myocardial stunning, low and high dose of tetrandrine groups. Another 10 rats were used to identify the calcium overload. vitamin D3 (0.3 million Unit/kg) and nicotinic acid were adm inistered. After 16 d when calcium overload occured, left anterior descending ar tery was ligated. Twenty minutes of myocardial ischemia followed by 60 min of re perfusion was induced. The contractile function parameters were determined dynam ically. At the end of experiment, myocardial cytosolic ［Ca2+］i was deter mined in various groups. In tetrandrine groups, tetrandrine (62.2 or 93.6 μmol/ kg ) was administered by gastrogavage daily.After 16 d, the rats undergone the e xperiments mentioned above. Results: Sixteen days after vitamin D3 , nicotinic acid were given, ［Ca2+］i increased by 2.6 folds (146.8±10.8 ) vs (368.5±22.6) nmol/L, (P<0.01). Whereas, ［Ca2+］i in tetrand rine groups were (210.8±16.4) and (198.6±15.3) nmol/L, which were significantl y lower than that of calcium overload group. Twenty minutes of myocardial ische mia resulted in the decrease of dp/dtmax and Vmax in all groups with the most si gnificant in stunning and calcium overload groups. The contractile function rest ored gradually after reperfusion. At all time points, dp/dtmax and Vmax in both tetrandrine groups were higher than those in both stunning and calcium overload groups. And effect with higher dose of tetrandrine were more significant than in low dose of tetrandrine. After 60 min of reperfusion, dp/dtmax in stunning, cal cium overload, low and high dose of tetrandrine groups were 49.7%, 51.5%, 71.0% and 83.4% of that in control, respectively， and Vmax were 55.0%, 49.8%, 73.9% and 77.5% of that in control, respectively. Conclusion: T he myocardial contractile function in vitamin D3-induced calcium overload gro up is impaired. On basis of myocardiocyte calcium overload, transient ischemia l eads to myocardial stunning. At the stage of ischemia, the impaired degree of my ocardial contractile function is similar to that in stunning group, suggesting a t this stage the effect of ischemia on myocardial function is greater than that of calcium overload. Tetrandrine chronically improves the myocardial function in Vitamin D3-induced calcium overload rats.