Experimental study of bone marrow-derived mesenchymal stem cells combined with hepatocyte growth factor transplantation via noninfarct-relative artery in acute myocardial infarction.

We investigated the impact of bone marrow-derived mesenchymal stem cells (BM-MSCs) alone or in combination with hepatocyte growth factor (HGF) transplantation via noninfarct-relative artery in a swine myocardial infarction (MI) model. Donor BM-MSCs were derived in vitro from swine auto-bone marrow cultures labeled by bromodeoxyuridine (BrdU) incorporation. Host MI swine model was created by ligating the distal left anterior descending artery. After 4 weeks, age-matched male MI swines were used for the transplantation. Male MI swines were transfused via noninfarct-relative artery with vehicle (control, n=6) or BrdU-labeled BM-MSCs (5 x 10(6)) alone (MSCs, n=6) or BrdU-labeled BM-MSCs (5 x 10(6)) combined with HGF (4 x 10(9) PFU) (MSCs+HGF, n=6). To evaluate the collateral artery growth (Rentrop) and cardiac perfusion in these animals, gate cardiac perfusion imaging and coronary angiography were performed before and 4 weeks after transplantation, respectively. To assess the contribution of donor-originated cells in stimulation of cardiomyocyte regeneration and angiogenesis, immunohistochemistry for BrdU and alpha-smooth muscle actin (alpha-SMA) and quantitative image analysis were performed at 4 weeks after transplantation. The results are as follows: (1) BrdU-positive cells were detected in host myocardium in both MSCs and MSCs+HGF groups, but not in the vehicle group. Most BrdU-positive cells expressed myosin heavy chain beta. (2) alpha-SMA(-)positive arteriole densities in the infarcted border area and infarcted area were increased significantly in both transplantation groups compared with the vehicle group. (3) Gate cardiac perfusion imaging demonstrated that the cardiac perfusion was significantly improved in transplantation groups compared with the vehicle group. (4) Ejection fraction and alpha-SMA-positive arteriole densities were increased significantly in both transplantation groups compared with the vehicle group. However, there was no difference in ejection fraction and alpha-SMA-positive arteriole densities between the MSCs group and the MSCs+HGF group. Growth of collateral arteries was not detected by coronary angiography in all three groups. In conclusion, the current study indicates that BM-MSCs transplantation via noninfarct-relative artery stimulates cardiomyocyte regeneration and angiogenesis and improves cardiac function, but does not stimulate collateral artery growth. BM-MSCs transplantation combined with HGF therapy is not superior to BM-MSCs alone transplantation. BM-MSCs transplantation via noninfarct-relative artery may be an alternative for those patients who cannot be transplanted via infarct-relative artery in clinical practice.

[Enzymatic conversion of cephalosporin C to glutaryl-7-aminocephalosporanic acid using whole cells of the yeast Trigonopsis variabilis FA10].

A process for the production of glutaryl-7-aminocephalosporanic acid (GL-7ACA) from cephalosporin C(CPC) using permeabilized cells of yeast Trigonopsis variabilis FA10 containing D-amino acid oxidase (DAO) is described. It was found that the bioconversion of CPC to GL-7ACA was interfered by the catalase activity presented in the cells that hydrolyzed the hydrogen peroxide and resulted in the accumulation of alpha-keto-adipyl-7-ACA (AKA-7ACA) and decrease of GL-7ACA yield. the methods to overcome this problem including the addition of extra H2O2 and use of catalase inhibitor, NaN3, were developed and the rate of GL-7ACA from CPC were 73% and 70.1%, respectively. Another alternative method was to incubate the permeabilized FA10 cells at pH10.5-11.0 for 30 minutes at 20 degrees C which served to selectively inactivate the catalase. In the bioconversion of CPC to GL-7ACA using pH10.5-treated cells without catalase activity, the high reaction yield of GL-7ACA(84%) was achieved.

[Permeabilization of yeast Trigonopsis variabilis FA10 cells for enhancing apparent activity of D-amino acid oxidase].

D-amino acid oxidase(DAO) is an intracellular enzyme in Trigonopsis variabilis FA10. The whole cells of yeast exhibited very low DAO activity. Various treatment with physical and chemical agents, such as, freezing and thawing, acetone, butanol and cetyltrimethylammoinium bromide altered membrane permeability and increased cellular DAO activity. It was demonstrated that the performance of acetone was dependent on the concentration of the solvent, the incubation time and the temperature. Maximum enzyme activity of the cells was achieved with 30-35% acetone, between 4 degrees C and 28 degrees C. The process was very quick and permeabilization occurred in 5 minutes or less. On the other hand, the thermostability of DAO in permeabilized cells was higher than in cell extract. It was proved that Cephalosporin C could be effectively converted to glutaryl-7ACA by permeabilized Trigonopsis variabilis cells with 83% yield.

An abduction-traction frame for the treatment of intertrochanteric fracture of femur.

We present a detailed outline of treatment of intertrochanteric fractures of femur with a new technique on the basis of principles of combined traditional chinese and western medicine. We use pins and an outer fixation frame, which keeps the injured limb in abduction and under traction and at the same time allows for functional exercise of all the joints in the extremity. The procedure is discussed and compared with other different techniques.

Comparison of effects of MPTCP and PTCP ceramics implanted for the treatment of bone defects in rabbits.

The magnetic porous tricalcium phosphate (MPTCP) and porous tricalcium phosphate (PTCP) ceramic cylinders were implanted into right and left bone defects of rabbits' radii in order to determine the utility of the MPTCP ceramics. Based on naked eye inspection, light and scanning electron microphotography, roentgenography, quantitative histological measurement of new bone formation and anti-break test for a period of 5 months. The results showed that the two kinds of ceramics were biocompatible with human tissue. MPTCP ceramics could induce more new bone formation than PTCP ceramics. Treatment of fractures with synthetic calcium phosphate ceramics and magnetic fields were discussed.

Macrophages in degradation of collagen/hydroxylapatite(CHA), beta-tricalcium phosphate ceramics (TCP) artificial bone graft. An in vivo study.

The macrophages mediated biodegradation of two biomaterials, collagen/hydroxylapatite (CHA) and beta-tricalcium phosphate ceramics (TCP), was studied in 24 male Kunming mice and 20 male C57BL/6 mice with histopathologic, histochemical and ultrastructural observation. It was demonstrated that macrophages infiltrated after CHA, TCP were implanted. The macrophages could be differentiated from fibroblasts and the other infiltrated cells for special cellular profile and strong acid phosphatase activity. Morphologically, monocyte-macrophages and infused multinuclear giant cell degraded CHA and TCP by phagocytosis and extracellular resorption. The carbonic anhydrase activity of macrophages was demonstrated by histochemical technique. It suggested that macrophages secreted H+ and accomplished the decalcification of calcium phosphate compound of CHA and TCP. We conclude that macrophages are the main mediating cells which degraded CHA and TCP intracellularly and extracellularly.

Macrophages in degradation of calcium phosphate compound artificial bone: an in vitro study.

The isolated mice peritoneal macrophages in degradation of calcium phosphate compound artificial bone--collagen/hydroxylapatite (CHA), hydroxylapatite (HA), beta-tricalcium phosphate (TCP) ceramics, have been studied by use of both Ca++, P concentration assay in cultured supernatant and scanning electron microscopy (SEM). The solubility of Ca++, composition of materials increased more significantly when macrophages were inoculated than when macrophages were not seeded (P < 0.001), and it was shown that the ground materials were wrapped and phagocytized or resorbed extracellularly by macrophages under SEM, suggesting that macrophages could mediate the degradation of calcium phosphate compound artificial bone by phagocytizing and/or degrading extracellularly.

Sonographic diagnosis of acute hematogenous osteomyelitis in the early stage.

This article presents sonographic diagnosis of acute hematogenous osteomyelitis in early stage. 24 children with clinically suspected acute hematogenous osteomyelitis were detected to have subperiosteal abscesses by ultrasound during four to fourteen days after onset. The mean length and anteroposterior distance of the subperiosteal abscesses were 86.4 mm and 10.7 mm, respectively. Of 24 cases of subperiosteal abscesses, aspiration performed under ultrasound guidance revealed purulent fluid in all and 23 were verified surgically. The results obtained indicate that ultrasound can be used in diagnosis of acute hematogenous osteomyelitis in the early stage. The earliest case was diagnosed by ultrasound 4 days after onset. By use of ultrasound, differentiation diagnosis of acute hematogenous osteomyelitis from other diseases such as cellulitis, soft tissue abscess, acute septic arthritis and malignant bone tumors is also discussed.

Ultrasound diagnosis of effusion of the hip.

This paper reports on two prospective studies of ultrasound examination of the hip in 150 healthy subjects and 50 patients with hip joint diseases. The sagittal distance of the anterior recess of the hip was measured on the sonogram of the sagittal section. The normal value in adults, teenagers and children was 4.54 mm +/- 0.99 mm, 4.27 mm +/- 1.26 mm and 3.40 mm +/- 0.93 mm, respectively. Ultrasound examination of both hips in 50 patients with hip joint diseases showed that the sagittal distance of the anterior recess was significantly widened in the symptomatic side, and the mean value in the symptomatic side and normal side was 12.02 mm +/- 3.68 mm, and 4.12 mm +/- 1.15 mm, respectively. Ultrasound showed echo-free area in anterior recess in 42 hips, hypoecho and medium echo area in 8 hips. Of 50 patients undergoing plain radiography, 33 showed bony changes of the hip, and the remaining 17 no change whatever. The results obtained demonstrated that ultrasound can accurately detect effusion of the hip, what is more, it is easy to operate, inexpensive, noninvasive, convenient for follow-up and without radiation damage.

Sonographic diagnosis of intramuscular hemangiomas.

This article reports 10 cases of intramuscular hemangiomas diagnosed by ultrasound. The results obtained demonstrated that the intramuscular hemangiomas were revealed sonographically as a spindle-shaped or ellipse-shaped mass with a mixed echo structure in the skeletal muscle, and usually with small calcifications. The solid parts of the tumor most commonly have a medium echogenicity, but few hyperechogenicity. The cavity or sinus containing blood has a hypoecho or echo-free structure.

New bone formation in the in vivo implantation of bioceramics. A quantitative analysis.

Two kinds of synthetic biomaterial, porous tricalcium phosphate (PTCP) and magnetic porous tricalcium phosphate (MPTCP) ceramic granules were implanted in rat femur. In the period of 4 months, the assessment of serial histological sections, scanning electron microphotographs and quantitative analysis of bone formation in the sections showed that both ceramics are biocompatible and degradable in vivo. More new bone formation occurred in the MPTCP group. Endochondral ossification was seen in both groups. The quantitative analysis in this study is reliable, and may be suitable to the similar experimental models.

Analysis of the biocompatibility of magnetic porous tricalcium phosphate ceramics in rat femurs.

A new biomaterial, magnetic porous tricalcium phosphate ceramics (MPTCP), may be used for repairing defects in bones as a substitute for bone grafting. This study was designed to examine its biocompatibility and the systemic toxicity. The ceramic sections, approximately 15.3 mg, were implanted into both proximal femurs of 10 male and 10 female albino SW rats. An equal number of male and female animals served as sham-operated controls. Implantation of MPTCP ceramics in rat femurs had no deleterious effects on body weight, organ/body weight ratio, and on blood, liver and kidney functions. Plasma levels of inorganic phosphorus were significantly elevated in rats implanted with MPTCP ceramics. Alkaline phosphatase values in plasma in the experimental group were equal to those in the control group. Photomicrographs, radiographs and scanning electron photomicrographs of the rat femurs implanted with MPTCP ceramics demonstrated excellent bone association with implants and growth of some new bone into MPTCP ceramics. In conclusion, MPTCP ceramics are biocompatible and suitable for the reconstruction of bones.

Artificial bone of porous tricalcium phosphate ceramics and its preliminary clinical application.

The authors have prepared the artificial bone of porous tricalcium phosphate ceramics according to an appropriate formula and manufacturing technology. Physical and chemical testing shows that it possesses several distinguishing features: the communicating pores and macro/micropores; mean pore size, 380 microns (from 240 microns to 510 microns); porosity, 46.4%; and compressive strength, 97.4 kg/cm2. It consists of CaO (49.09%) and P2O5 (48.84%). The testing of its biocompatibility shows that it is devoid of systemic or local toxicity, and free of irritation or foreign body response in tissues, and it does not result in hemolysis or mutation. The new bone readily grows into its pores with direct contact to the implanted material. 11 cases of bone defects were treated with this artificial bone with satisfactory results.