Objective To investigate the influence of low frequency micromovement induced by an external fixator with controlled micromovement device on callus formation and fracture healing. Methods 39 sheep were involved in the study. After transverse osteotomy with a gap of 2 mm was done at the mid-shaft of both tibias, the hind limbs were fixed with unilateral external fixators with controlled micromovement device. 10 days after osteotomy, one hind limb of the sheep was randomly selected to have micromovement. According to different micromovement frequencies, the sheep were randomly divided into 3 groups: Group A, 0.5 Hertz; Group B, 1 Hertz; Group C, 5 Hertz. The micromovement had an amplitude of 0.25mm and stopped by the end of the fourth postoperative week (30 min/d). The other hind limb of each sheep did not have micromovement and served as the control. Macrostructural and histological observations, radiographic examinations, and biomechanical tests were done respectively at 4, 6, 9 weeks after osteotomy. Results Macrostructural observations and X-ray examinations revealed a large amount of callus formation with a peak at the end of the fourth postoperative week in the 3 experimental groups. Histological analysis showed faster endochondral ossification and osteoid formation in the experimental groups than in the control by the sixth postoperative week, with the highest rate in Group B (1Hz). Biomechanical tests proved that the bending stiffness in the experimental groups was superior to that in the control, with the strongest in Group B (1 Hz). Conclusions Low frequency-micromovement at the fracture sites can promote callus formation and accelerate mineralization at the early stage of fracture healing. A frequency of 1 Hz can produce the best results.

Objective To obtain the Tat-GFP fusion proteins with penetrating activity and labeled with green fluorescence protein (GFP), and to explore the cell membrane penetrating activity of Tat-GFP in MCF-7 cells. Methods The plasmid pET-24a-Tat-GFP was transformed into Escherichia coli BL21 cells. Different concentrations (0.5 and 1.0 mmol · L-1 ) of isopropyl-β-D-thiogalactopyranoside (IPTG ) and cell culture temperatures (22℃ and 37℃)were used to optimize the protein expression.The Tat-GFP proteins in supernatant were purified using Ni-IDA resins. Western blotting analysis was used to identify the Tat-GFP protein, and confocal laser scanning microscope (CLSM ) was used to examine the cell penetration of Tat-GFP protein. Results There was no significant difference in the Tat-GFP protein production induced by 0.5 and 1.0 mmol·L-1 IPTG;however,the low temperature (22℃)-induced BL21 cells expressed more Tat-GFP proteins than that at 37℃ induction.The Western blotting analysis results showed that GFP antibody could specifically recognize the proteins in PVDF membranes in dose-dependent manner;the CLSM results indicated the distribution of green fluorescence in cytoplasm and nucleus of MCF-7 cells.Conclusion The Tat-GFP protein highly expresses in the supenatant of Escherichia coli i BL2 1 cells at low temperature;the obtained Tat-GFP protein with green fluorescence preserves the cell penetrating activity.

Objective To investigate the kinesin family member 17 (KIF17) expression and cellular localization in the hippocampus and temporal lobe cortex in the rat lithium-pilocarpine model of epilepsy,and discuss its function in the epilepsy pathogenesis.Methods The animal model was established by lithiumpilocarpine induction in rats.Totally 49 adult healthy male Wistar rats were randomly divided into control group (n =7) and experimental group (n =42).The experimental group included 6 subgroups (n =7)according to sacrifice time (24 h,72 h,7 d,14 d,1 month and 2 months).The expression and localization of KIF17 were examined by western blot and double-label immunofluorescence,respectively.Results In rat hippocampus,the expression of KIF17 protein increased after the onset of seizure (the ration of KIF17/β-actin were:24 h 0.516 ± 0.196,72 h 0.742 ± 0.313),reached its peak in 7 days (0.888 +0.319)and then slowed down (14 d 0.770 ± 0.271,1 month 0.742 ± 0.261,2 months 0.714 ± 0.271),all of which were significantly higher than that in the control group (0.495 ± 0.203).And all the differences had statistical significance (t =7.051,4.974,7.419,8.795,8.264,6.676,all P ＜ 0.05).In rat cortex of temporal lobe,the expression of KIF17 protein increased after the onset of seizure and reached its peak in 30 d.The optical density ration in the experimental groups were higher than that in the control group.Doublelabel immunofluorescence disclosed that the KIF17 localized in the neurons,including excitable neurons and inhibitory neurons,but not in the astrocytes which were performed with anti-microtubule-associated protein 2,anti-brain-specific Na-dependent inorganic phosphate cotransporter,anti-glutamate decarboxylase 1 and anti-glial fibrillary acidic protein,respectively.Conclusion KIF17 may play a potential role in the pathogenetic mechanisms of the rat lithium-pilocarpine model of epilepsy.

Objective To evalute the effect of Ilizarov technique in the treatment of hypertrophic nonunion.Methods Form June 2008 to December 2010,12 patients with hypertrophic nonunion were treated with Ilizarov technique,including 10 males and 2 females with an average age of 46.5 years.The pathology sites of nonunion were kept as closed as possible without any bone graft during operation.As to patients who had ever been treated with plate or intramedullary nail,the hardware should be removed by minimal invasive approach.These procedures aimed to keep the vascularity of nonunion site intact.Ilizarov apparatus were preoperatively constructed.Distal segment and proximal segment of nonunion were mounted respectively with two external circle using the smooth wires and half pins.The two-circle stabilizing one segment was nominated with transosseous modules.Distal module and proximal one was connected with a pair of axial hinges.The pathology sites were gradually distracted from the seventh day postoperatively,0.25 mm/d.Accompanying with deformity correction,limb length discrepancy (LLD) also were restored simultaneously.Then,all the screws and nuts in the apparatus should be tightened,which was favourable to the callus consolidation.Results All 12 cases of nonunion healed without any bone graft.The fixator wearing time lasted 6-12 months,with an average of 8 months.Correction of deformity and LLD were achieved.The average lengthening was 3.0 cm (range,2.0-5.5 cm),the average correction angle was 23° (range,10°-30°).After 6-18 months follow-up,all the patients restored satisfactory function.Conclusion Hypertrophic nonunion can be treated successfully with Ilizarov technique.The key of successful callus distraction is strictly identifying the indications.

Objective To investigate the copy number changes on chromosome 3q26. 1 in urothelial carcinoma of the bladder, and to explore its potential clinical significance. Methods The microarray-based comparative genomic hybridization (Array-CGH) approach was used to analyze the genome-wide copy number changes of 35 tumor tissue samples of bladder cancer. To confirm the loss of a small fragment in 3q26. 1 detected by Array-CGH, real-time fluorescent quantitative polymerase chain reaction (real-time PCR) was performed with 57 frozen tumor tissue samples and 34 formalinfixed paraffin-embedded (FFPE) tumor tissue samples. The urine sediment cells collected from 15 healthy volunteers and 29 bladder cancer patients were checked as above. Results The Array-CGH data showed that the copy number loss of a small fragment in 3q26. 1 was detected in 77.1% (27/35)of the tumor tissue samples investigated. Real-time PCR analysis validated this loss of a small fragment of 3q26.1 with high frequencies in both 57 frozen tumor samples and 34 FFPE tumor samples.The percentage of samples exhibiting loss was 78.9% (45/57) and 100. 0% (34/34) respectively.Furthermore, the relative copy number of the 3q26.1 small fragment was significantly lower in the urinary sediment cells of the patients (median=0. 0020), comparing with that of healthy controls (median=0. 0030) (P＜0.01). Conclusions Loss of the small fragment in 3q26.1 could be a characteristic genetic change of urothelial carcinoma of the bladder. It may serve as a potential molecular marker for bladder cancer.

Objective To study the effects of one finger massage on sciatic nerve injury rats. Methods The sciatic nerves were exposed, and the sciatic nerve was held by micro needles to make the sciatic nerve injury model. SD rats were randomly divided into sham-operation group, model group and massage group. In the sham-operation group, the sciatic nerves were exposed but not held. 7 d after the establishment of modeling, rats in massage group received one finger massage for 30 d. After 30 d, SFI and BBB of sciatic nerves were detected. HE staining, transmission electron microscope and immunochemistry assay were used to measure the changes of sciatic nerves. Results Compared with model group, massage group could speed up the recovery of SFI and increase BBB, promote the recovery of sciatic nerve morphology, increased protein level of S-100β, and enhance ultrastructure of newborn nerve growth and recovery. Conclusion One finger massage can effectively promote neurological and functional recovery after sciatic nerve injury in rats.

Wound healing is a complex and sequential biological process involving various cells and factors under body′s regulation. Appropriate interventions play positive roles in promoting effective wound healing and improving healing quality. In the clinical practice, there are many new instruments, dressings, and drugs developed for wound care, including antibacterial dressings, wet dressings, precise debridement, negative pressure wound therapy, cytokines/growth factors, and dermal substitutes, which provide revolutionary means for wound treatment. This article summarizes the effective or mature methods in wound care, providing theoretical and practical basis for choosing appropriate treatment methods in different stages of wound.

Epithelial membrane protein 3 (EMP3) is a trans-membrane signaling molecule with important roles in the regulation of apoptosis, differentiation and invasion of cancer cells, but the detailed is largely still unknown. We analyzed the mRNA levels and methylation statuses of EMP3 in 63 primary breast carcinomas and assessed their correlations with clinicopathologic variables. The expression of EMP3 mRNA in primary breast carcinomas was significantly higher than the expression of 20 normal breast tissues (p<10(-7)). EMP3 overexpression in breast carcinomas was significantly related to histological grade III (p=3.9X10(-7)), lymph node metastasis (p= 0.003), and strong Her-2 expression (p=3.3X10(-6)). Hypermethylation frequencies of EMP3 were detected in 36.5% of breast carcinomas by methylation-specific polymerase chain reaction. However, no significant correlations were found between methylation status of EMP3 and mRNA expression levels as well as other clinical parameters. In conclusion, EMP3 may be a novel marker of tumor aggressiveness. Overexpression of EMP3 in primary breast carcinoma is not associated with DNA methylation.
Adult ; Breast Neoplasms/*genetics/pathology ; Carcinoma/*genetics/pathology ; DNA Methylation ; Epigenesis, Genetic ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Lymphatic Metastasis ; Membrane Glycoproteins/*genetics/metabolism ; Middle Aged ; Neoplasm Staging ; RNA, Messenger/metabolism ; Receptor, erbB-2/genetics/metabolism ; Severity of Illness Index

This is a work aimed to investigate the biodegradability, biocompatibility and mechanical property of the poly-lactide-co-glycolic acid (PLGA) knitted mesh preliminarily and to further explore its applications in tissue engineering and regenerative medicine. The biological property of PLGA mesh was investigated comprehensively with the degradation experiment in vitro, the acute cytotoxicity assay, the intradermal irritation test and the subcutaneous implantation test in vivo utilized. The degradation experiment in vitro demonstrated that the pH value of the removed solution fluctuated between 6.68 and 7.33. The elastic modulus of the PLGA mesh increased at first and then decreased afterwards. The acute toxicity test and the intradermal irritation test indicated that the PLGA mesh was with innocuity safety. The PLGA mesh accelerated degradation and was replaced gradually by the neotissue. The results of immunohistochemical staining demonstrated that the number of ED-1+ cells increased at first and then decreased afterwards. The PLGA mesh with excellent mechanical properties, good biocompatibility and favorable degradation ratio has the potential to be employed as a "skeleton" to reinforce the mechanical property of collagen-based dermal substitutes in tissue engineering.
Absorbable Implants ; Biocompatible Materials ; chemistry ; Humans ; Lactic Acid ; chemistry ; Materials Testing ; Polyglycolic Acid ; chemistry ; Skin, Artificial ; Tissue Engineering ; methods