OBJECTIVETo investigate the optimum proportion of Mosaicplasty and genes-enhanced tissue engineering for the repair of acute osteochondral defects.

METHODSWestern blot test was conducted to detect the expression of hTGF-beta1, Col II and Aggrecan in 3 groups, including hTGF-beta1, transduction group, Adv-betagal transduction group and control group without transduction. Eighteen 6-month-old Shanghai male goats (weight: 22 to 25 kg) were used. BMSCs were isolated from the autologous bone marrow, and were subcultured to get the cells at passage 3. Thirty-six medial femoral condyles were used and divided into 6 groups named AR, AL, BR, BL, CR, and CL. Acute cylindrical defects (9 mm in diameter and 3 mm in depth)were created in the weight bearing area of the medial femoral condyle of hind limbs. In the single group, the autologous osteochondral mosaicplasty was performed to repair the defect; in the combination group, besides the mosaicplasty, the dead space between the cylindrical grafts and the host cartilage were injected with the suspension of hTGF-beta1, gene enhanced autogenous BMSCs in sodium alginate, and CaCl2 was dropped into it to form calcium alginate gels. The autologous osteochondral transplantation cover rates of group AR was 44.44% single group, AL was 44.44% combination group, BR was 33.33% single group, BL was 33.33% combination group, CR was 22.22% single group, and CL was 22.22% combination group. The goats were killed 24 weeks after operation to receive gross and histology observation, which was evaluated by the histological grading scale of O'Driscoll, Keeley and Salter. Immunohistochemistry and TEM observation were also performed.

RESULTSWestern blot test showed the expression of the hTGF-beta1, Col II and the Aggrecan in the hTGF-beta1 transduction group were significantly higher than that of the Adv-betaga1 transduction and the blank control groups. The gross and histology observation revealed that each defects of six groups had different degrees of repairing. There was no significantly difference among the BL, AR, and AL groups. But the scores of the other three groups (BR, CR, and CL) were significantly poorer than the former three groups.

CONCLUSIONMosaicplasty associated with genes enhanced tissue engineering could repair the osteochondral defects effectively. With the autologous osteochondral transplantation coverage reducing, the advantage of the combination could have a better representation.

Animals ; Blotting, Western ; Bone Diseases ; metabolism ; pathology ; therapy ; Cell Line ; Goats ; Humans ; Immunoprecipitation ; Male ; Tissue Engineering ; methods

OBJECTIVETo investigate whether the unevenness of articular surface would affect the osteochondral repair.

METHODSEight Shanghai Chongming 6-months-old masculine goats with a mean weight of 25 kg were used in this study. Different unevenness, which were 0.5 mm, 1.0 mm, 2.0 mm protrude or concavity, were created on the weight-bearing portion of the medial femoral condyles of the goats. The goats were sacrificed 12 weeks later and were observed with the general observation, HE staining and transmission electron microscope. To evaluate the microscopic morphology, a histological grading scale described by O'Driscoll, Keeley and Salter was used.

RESULTSThe general observation and HE staining showed that the unevenness of 0.5 mm or 1.0 mm protrudes or concavity could be repaired to get the surface smooth on the whole. The transmission electron microscope showed that the reparative tissues were the same as the normal cartilage. The 2.0 mm depth couldn't be repaired satisfactorily. The transmission electron microscope showed that the fiber bundle proliferated and the chondrocytes degenerated. The scores of the 2.0 mm depth were significantly lower than that of the 0.5 mm or 1.0 mm (P < 0.05).

CONCLUSIONThe unevenness could have an influence on the repair. The limited unevenness could be repaired by itself.

Animals ; Cartilage, Articular ; pathology ; surgery ; ultrastructure ; Male ; Microscopy, Electron, Scanning ; Sheep ; Wound Healing

To analyze the relevant documents of hospital infection administration of acupuncture manipulation with filiform needles and acupuncture aseptic technique with filiform needles. The current situation is that acupuncturists have understanding insufficiency in hospital infection management, lack the sterile concepts and consciousness of disinfection and isolation. Aseptic technic principles aren't strictly followed; disinfection and isolation systems are unsound; sanitary condition of hand of medical staff is unsatisfied; and there is shortness in traditional long filiform needle manipulation. In future, we should explore the new model of hospital infection administration of acupuncture manipulation with filiform needles from implementations of relevant rules of hospital infection administration, establishment and supervision of sound corresponding system, further research of manipulation of filiform needles and formulation of septic technic criterion of filiform needles.
Acupuncture Therapy ; instrumentation ; standards ; Cross Infection ; prevention & control ; Disinfection ; standards ; Equipment Contamination ; prevention & control ; Hospital Administration ; Humans ; Hygiene ; standards ; Needles

OBJECTIVETo study the evaluation of bipolar radiofrequency (RFE) chondroplasty to cartilage injure.

METHODSSixteen goats underwent resection of anterior cruciate and medial meniscus to create cartilage injured model. Bipolar frequency energy chondroplasty were performed on injured articular cartilage 3 months after operation. The left knee was regarded as sham-operated control. The zero time effects and later changes of radiofrequency on cartilage surface and chondrocytes were observed.

RESULTSAt zero time, the rough injured surface became smooth, and clefts were melted. Some chondrocytes were dead in the superficial layer, cells in deeper layer remained alive. Three months later, the surface of the articular was smoother than the sham-operated side. The histological modified Mankin' s score of cartilage was significantly higher than the sham-operated side (operated side 12 +/- 4; sham-operated side 14 +/- 5, P < 0.01). The result of Fluorescence-activated cell sorter (FACs) showed that there were no obvious difference of dead chondrocyte between operated side and sham-operated side, (24 +/- 10)% and (12 +/- 1)% (P > 0.05).

CONCLUSIONSBipolar radiofrequency energy (1 grade) does not create more chondrocyte death than the sham-operated side. Bipolar radiofrequency chondroplasty is an appropriate method to treat articular cartilage injury.

Animals ; Cartilage, Articular ; injuries ; pathology ; surgery ; Catheter Ablation ; Chondrocytes ; pathology ; Disease Models, Animal ; Female ; Goats ; Male

OBJECTIVETo investigate the effectiveness of human insulin-like growth factor I (hIGF-I) gene transferred into the cultured goat bone marrow mesenchymal stem cells with liposome, and find a new method of cell-mediated gene therapy.

METHODSBone marrow was extracted from adult goats and cultured in vitro by monolayer. Then the recombinant eukaryotic expression plasmid pIRES2-EGFP-hIGF was transfected into cells by FuGene 6. After transfection, the marker gene coding enhanced green fluorescent protein (EGFP) was observed at different time points. The hIGF concentration in the supernatant fluids was measured by enzyme-linked immunosorbent assay (ELISA) and immunohistochemistry stain of hIGF was performed to detect the target protein. Besides, reverse transcription polymerase chain reaction and flow cytometry were also adopted in order to find out the changes of cells after transfection.

RESULTSBone marrow stem cells were all in the long shuttle-like shape and adhered to the disk. The expression of EGFP was first found at 4 h after transfection. The amount and intensity of EGFP increased gradually during the period of detection and got to the peak degree at 72 h, after that decreased slowly. EGFP was also seen in the second generation cells, but the intensity was relatively faint. The IGF-I concentration secreted into the supernatant was in accordance with the EGFP observed with the peak concentration at 34.75 ng/ml. The outcome of RT-PCR and immunohistochemistry was positive. The morphology of many stem cells was changed into triangular or irregular forms under the circumstance of the secreted hIGF-I. Percentage of stem cells in the S stage increased after transfection.

CONCLUSIONThe hIGF-I gene can be transfected efficiently and safely into BMSCs by FuGene 6, and the hIGF-I protein can be secreted into the supernatant in a relatively high level during a long period, therefore accelerate the proliferation and differentiation of the transfected cells.

Animals ; Bone Marrow Cells ; cytology ; Cell Differentiation ; Genetic Vectors ; Goats ; Humans ; In Vitro Techniques ; Insulin-Like Growth Factor I ; biosynthesis ; genetics ; Male ; Mesenchymal Stromal Cells ; cytology ; Plasmids ; genetics ; Transfection

OBJECTIVETo construct a reasonable substitute for the autograft bone in vitro and transplant it back into the rabbit models to induce the spine fusion.

METHODSThe bone marrow stem cell from the seven New Zealand rabbits were cultured. Recombinant human bone morphogenetic protein-4 (rhBMP-4) that has been proved to be bioactive was obtained by the way of genetic engineering. Using the vacuum freezing machine to mix a certain quantity of rhBMP-4 into type I collagen to form a new kind of carrier. Animal model of spine facet process fusion was used. Bone marrow stem cells combined with rhBMP-4 and I type collagen were implanted between the facet process to induce the spine fusion. type I collagen and bone marrow stem cell was used in the controlled group.

RESULTSNew bone formation was obvious in the test group. The facet joint was fused very well in this side. No bone formation was present on the other side.

CONCLUSIONSThe new composite: bone marrow stem cells, rhBMP-4 and type I collagen was an ideal kind of substitute for the autograft bone.

Animals ; Bone Marrow Cells ; cytology ; Bone Morphogenetic Protein 4 ; Bone Morphogenetic Proteins ; biosynthesis ; genetics ; Bone Substitutes ; therapeutic use ; Bone Transplantation ; Cells, Cultured ; Collagen Type I ; chemistry ; Implants, Experimental ; Rabbits ; Recombinant Proteins ; biosynthesis ; genetics ; Spinal Fusion ; methods ; Stem Cell Transplantation ; Stromal Cells ; cytology ; Tissue Engineering

OBJECTIVEHerein, the synthesis, component, microstructure and pharmacological and toxicology researches of the Synthetic Mercury Sulfide (S-HgS) a kind of common drug in Chinese, Mongolia, Tibetan medicine, and Indian medicine system were summarized. The similar cognition about mercury toxicity & pharmacological action from some Asian regions was analyzed, and it can supply some useful direction for the traditional Asian medicine system.

METHODRecent literatures both domestic and abroad were summarized and analyzed.

RESULTS-HgS is the basis of Vermilion, Mongolia-Vermilion, Zuotai, and Ras-sindoor. Athough the processes of synthesis are very different, but the microstructure and pharmacological & toxicology of S-HgS is similar.

CONCLUSIONS-HgS has a far-ranging application,and unique curative effect. New technology such as nanotechnology can be used for improving the advancement of traditional Asian medicine.

Humans ; Medicine, Traditional ; Mercury Compounds ; adverse effects ; chemistry ; pharmacology ; Sulfates ; adverse effects ; chemistry ; pharmacology

OBJECTIVETo establish of acute respiratory distress syndrome (ARDS) model in canine after inhalation of perfluoroisobutylene (PFIB), and to observe the progressing of lung injury, and to study the mechanisms of injury.

METHODSA device of inhalation of PFIB for canine was made. The concentration of PFIB was 0.30 - 0.32 mg/L. Serum IL-6 and IL-8 were dynamically measured. Clinical manifestations, pathology of organs in canine were observed.

RESULTS(1) During inhalation, the concentration of PFIB remained stable; (2) After inhalation, blood arterial oxygen partial pressure fell gradually, and eventually met the criteria for diagnosing ARDS; (3) The level of IL-8 in serum rises significantly after inhalation (P < 0.05), whereas that of IL-6 was not obviously altered (P > 0.05); (4) Within 6 hours after inhalation, no abnormality in canine was observed, but afterwards symptoms gradually appeared, and typical breath of ARDS, such as high frequency and lower level could be seen in later phase; (5) Pathological examination showed severe congestion, edema and atelectasis in most part of both lungs, and signs of anoxia in other organs.

CONCLUSIONS(1) The device designed is capable of ensuring control of inhalation of PFIB; (2) Exposure to PFIB for 30 mins, canines all met the criteria for diagnosing ARDS 22 hours after inhalation, therefore the modeling is successful; (3) PFIB specifically damages the lung by causing excessive inflammation.

Administration, Inhalation ; Animals ; Disease Models, Animal ; Dogs ; Female ; Fluorocarbons ; toxicity ; Interleukin-6 ; blood ; Interleukin-8 ; blood ; Lung ; drug effects ; pathology ; Male ; Random Allocation ; Respiratory Distress Syndrome, Adult ; blood ; chemically induced

BACKGROUNDChromosomal abnormalities have been shown to play an important prognostic role in multiple myeloma (MM). Interphase fluorescence in situ hybridization (i-FISH) has been much more effective to identify cytogenetic aberrations in MM than conventional cytogenetic technique (CC). To clearly determine the cytogenetic features of Chinese MM patients and identify their prognostic implications, we designed a multicenter study based on i-FISH including 672 patients from 52 hospitals in China.

METHODSAll 672 patients were systematically screened for the following genomic aberrations: del(13q), IgH rearrangement, del(p53) and 1q21 amplifications.

RESULTSThe analysis showed that the chromosomal changes were detected in 22.1% patients by CC and in 82.3% patients by i-FISH. The most common abnormalities by CC were chromosome 1 aberrations (48.4%), -13/13q- (37.6%), hyperdiploidy (36.6%), hypodiploidy (30.1%) and IgH rearrangements (23.7%). The most frequent abnormalities by FISH was del(13q), which was found in 60.4% patients, whereas IgH rearrangement, 1q21 amplification and p53 deletions were detected in 57.6%, 49.0% and 34.7% cases, respectively. By statistical analysis, -13/13q- by CC was associated with low level of platelet (P = 0.015), hyperdiploidy was associated with low level of serum albumin (P = 0.028), and IgH rearrangement by FISH was associated with high level of β2 microglobulin (P = 0.019). Moreover, 1q21 amplification and del(p53) by FISH conferred a high incidence of progressive disease (PD) after initial therapy. Metaphase detection of IgH rearrangements and chromosome 1 aberrations concurrently was associated with a short progression free survival (PFS) (P = 0.036). No significant prognostic implications of other cytogenetic abnormalities were found associated with overall survival and PFS.

CONCLUSIONSChinese MM patients had similar cytogenetic abnormalities compared with the previous reported studies. However, the prognostic significance of FISH aberrations were not clearly determined and further study is required.

Adult ; China ; Chromosome Aberrations ; Chromosomes, Human, Pair 1 ; genetics ; Cytogenetic Analysis ; Female ; Humans ; In Situ Hybridization, Fluorescence ; Karyotyping ; Male ; Middle Aged ; Multiple Myeloma ; genetics ; pathology