BACKGROUND: The survival rate of articular chondrocytes is low after traditional cryopreservation,and great differences existed in chondrocytes from surface layer and deep layer,which easily result in graft degeneration and lead to surgery failure.OBJECTIVE: To establish rabbit allograft models of graded frozen articular cartilages with holes made before cryopreservation and to observe the effect of holed cryopreservation on the rabbit articular cartilages.METHODS: Osteochondral plugs taken aseptically from 2 months old rabbits were randomly divided into 3 groups: the experimental group,making holes(3 mm× 3 mm)in articular cartilages and graded freezing; non-hole graded freezing group,non-making holes and graded freezing; cryopreservation group: non-making holes and rapid freezing.The grafts were thawed and transplanted into the relevant articular cartilage defects of recipient rabbits.The grafts differences were observed by gross observation,histochemistry and immunohistochemistry staining.RESULTS AND CONCLUSION: The gross observation,histochemistry and immunohistochemistry staining of the experimental group were superior to the cryopreservation group.Though there were no significant differences between the non-hole graded freezing group and the experimental group,however,the experimental group enhanced the protective effect on cartilage tissue in the middle layer.The graded cryopreservation of articular cartilage gets an advantage over rapid cryopreservation.And the articular cartilage with holes could be preserved successfully in graded cryopreservation,which assures the survival and function of chondrocytes and slows down degrading process of the articular cartilage tissue after thawed and transplanted.

OBJECTIVETo analyze the effects of metacarpus and phalanx traction on correction of scar contracture of hand after burn on the palm side retrospectively.

METHODSA total of 32 patients with 39 affected hands with scar contracture on the palm side after burn were hospitalized from May 2010 to December 2014. Method of treatment: scar contracture was conservatively released followed by skin grafting, which was referred to as method A; Kirschner wire was inserted into the middle or distal phalanx of finger with contracture and the corresponding metacarpus in the shape of U for 2 to 7 weeks' traction, which was referred to as method B; traction frame was built based on the traction pile and anchor formed by Kirschner wire inserted through the second to the fifth metacarpus and distal phalanx of finger with contracture, and then the affected fingers were pulled into a straight position with rubber bands for 2 to 6 months, which was referred to as method C. Method A was used in patients who would be treated with thorough release of scar followed by skin grafting routinely. Method B was used in patients who would be treated with intramedullary Kirschner wire fixation after release of scar contracture and skin transplantation routinely. Method C was further used in patients when methods A and B failed to accomplish the expected result. Method C was used in the first place followed by method A in whom there might be vascular decompensation or exposure of tendon and bone after scar release, and those who failed to meet the expectation were treated with method C in addition. Patients who were unwilling to undergo surgery were treated with method C exclusively. During the course of treatment, the presence or absence of infection and slipping of Kirschner wire or its slitting through soft tissue were observed. The presence or absence of tendency of recurrence of scar contracture within 1 to 2 weeks after treatment was observed. The length of palmar skin measuring from the root of finger with contracture to wrist crease was measured before treatment, at the termination of treatment, and 1 month after the termination of treatment. Scar condition was assessed with the Vancouver Scar Scale (VSS) before treatment and 1, 3, and 6 month(s) after the termination of treatment. Before treatment and 1 month after the termination of treatment, the range of motion was measured with the Total Active Movement (TAM) method; band function was evaluated by the Jebsen Test of Hand Function (JTHF), and the completion time was recorded. Data were processed with analysis of variance, LSD-t test, and t test.

RESULTSTwenty-four patients with 27 affected hands were treated with scheme A + B; 5 patients with 7 affected hands were treated with method C exclusively; 2 patients with 3 affected hands were treated with scheme A + B + C; 1 patient with 2 affected hands were treated with scheme C + A + C. During the course of treatment, no complication such as infection or slicing of tissue was observed, but there was a slight shifting of U-shaped Kirschner wire in 14 affected hands of 13 patients. Tendency of recurrence of scar contracture was observed in 11 affected hands of 10 patients, but the scar contracture did not reoccur after treatment with orthosis. The skin length of palmar side was respectively (131.8 ± 9.8) and (127.6 ± 7.5) mm at the termination of treatment and 1 month after, and they were both significantly longer than that before treatment [(114.5 ± 2.4) mm, with values respectively 10.71 and 10.39, P values below 0.001]. The score of VSS was respectively (9.8 ± 2.4), (9.7 ± 1.7), (9.3 ± 0.8), and (7.7 ± 0.5) points before treatment and 1, 3, and 6 month(s) after the termination of treatment. Only the score at 6 months after the termination of treatment was significantly lower than that before treatment (t = 3.28, P < 0.01). The ratio of excellent and good results according to method TAM was respectively 2.6% (1/39) and 94.9% (37/39) before treatment and 1 month after the termination of treatment. The time for JTHF measurement was (13.9 ± 4.1) min before treatment, and it was shortened to (11.0 ± 2.8) min 1 month after the termination of treatment (t = 3.65, P < 0.001).

CONCLUSIONSSingle application of metacarpus and phalanx traction or its combination with skin transplantation after scar release in correcting scar contracture of the palm of hand after burn can lengthen the contracted tissue, and it is beneficial for the restoration of function and appearance of affected hand.

Burns ; rehabilitation ; therapy ; Cicatrix ; therapy ; Contracture ; surgery ; Hand Injuries ; rehabilitation ; therapy ; Humans ; Metacarpus ; Orthotic Devices ; Range of Motion, Articular ; Reconstructive Surgical Procedures ; methods ; Retrospective Studies ; Skin ; Skin Transplantation ; Tendons ; Time ; Traction ; Treatment Outcome

The aim of this study was to build a gene chip system with surface plasmon resonance (SPR) technique, for which Gamma-peptide nucleic acid (Gamma-PNA) functioned as a probe, in order to improve sensitivity and its specificity. With the use of self-assembled monolayer (SAM) technology, surface chemistry of two-dimensional structure was used. Gamma-PNA was designed according to the bioinformatics, and was plated on the SPR chip modified by SAM. Subsequently, relevant parameters of the experiment were ensured and optimized. The results showed that the performances of Gamma-PNA probe was little affected by the ion concentration of buffer, and it had a strong light signal in a stable state. As the ion concentration was 0, there were still good hybrid reactions; pH value had less influence upon Gamma-PNA probe, and acid environment of buffer could be better. Gamma-PNA probe combined with sensor technologies achieved made the probe with dispensable labels and real-time detection. It also improved the efficiency of the hybridization and the stability, providing the foundation for clinical application.
Nucleic Acid Hybridization ; Nucleic Acid Probes ; Oligonucleotide Array Sequence Analysis ; methods ; Peptide Nucleic Acids ; genetics ; Sensitivity and Specificity ; Surface Plasmon Resonance

The cyp51 primers and two pairs of mutant primers which removed different transmembrane region were designed based on Ustilago maydis cyp51 gene structure analysis. The full cyp51 DNA fragment as well as mutant cyp51 genes were amplified and cloned by using the total DNA from Ustilago maydis as template, then subcloned into different expression vectors. The recombinant expression plasmids were transformed into Escherichia coli BL21 (DE3), BL21 (DE3) pLysS and Rosetta (DE3) respectively. A series of experiments leads to the finding that only pET32-YH-35 could be highly expressed at the optimal condition of 30 degrees C induced with 0.5 mmol/L IPTG The expressed protein (CYP51) showed biological activity by spectra analysis of the protein binding to 4 standard fungicides and to 14 XF-synthetic fungicide compounds, and only one XF-synthetic fungicide compound (XF-113) was similar to standard fungicides in binding constant. This compound is promising to be a new effective antifungal drug. These results will facilitate the further study on the mechanism of pathogenic fungi CYP51 and pesticide molecules, and will provide a new idea for efficient design and development of new anti-fungal drugs.
Antifungal Agents ; isolation & purification ; Cloning, Molecular ; Cytochrome P-450 Enzyme System ; genetics ; metabolism ; Escherichia coli ; genetics ; metabolism ; Fungal Proteins ; genetics ; metabolism ; Plasmids ; genetics ; Recombinant Proteins ; genetics ; metabolism ; Sterol 14-Demethylase ; Ustilago ; enzymology ; genetics