Objective To explore a better method for treatment atrophic rhinitis.Methods 56 patients with atrophic rhinitis(96 lateral)were treated by nasal submucou pediculated bone-suberiosteal muscle flap extracted from anterior wall of sinus maxillaries.Results All patients were followed 2 to 10 years,total effective rate was 100 %, with 49 cases(87.5 %)showing prominent effect.Conclusion The grafted flap cannot be assimilated,felled off and necrosis,because the flap has rich blood supply.This methods has obvious short-term effective and stable long-term effective.No complications were found.

Objective To investigate the efficacy of the bioactive artificial vertebrae of a nano- hydroapatite crystals and polyamide 66 composite(n-HA/PA66)to restore the height and architecture of thoracolumbar burst fracture.Methods From December 2003 to February 2006,38 patients(29 males and 9 females)with a mean age of 35.6 years(17-63 years)were treated surgically through anterior ap- proach for decompression and implanted with the bioactive artificial vertebrae of n-HA/PA66 composite to reconstruct the structure of the thoracolumbar burst fractured vertebra.Results All the patients were successfuly followed-up for an average of 8 months,ranging from 6 to 21 months.The bioaetive artificial vertebrac of n-HA/PA66 composite were fused with the receptor bone 3-4 months after operation.The neu- rological function of the patients was restored partially or completely.The thoracolumbar spine was stable during physical examination and the height of thoraeolumbar burst fractured vertebrae that had been restored did not changa during the follow-up.Conclusions Our results show the bioaetive artificial vertebrae of n-HA/PA66 can restore the height and structure of thoracolumbar burst fractured vertebrae and reconstruct the structure of the tboraeolumbar vertebrae effectively,indicating that the bioaetive artificial vertebrae of n- HA/PA66 can be used extensively in clinical spinal surgery.

OBJECTIVETo observe the effects of the fat flap tissues after delay operation on free fat-graft survival rate and duration.

METHODSThe delay operation of fat flaps was performed in the inguinal region of a rabbit. Expression of VEGF was assayed using Elisa method after 12 hours of flap delay. The fat flaps were harvested and cut into pieces after 21 days. A subdermal pocket was created in each side of the dorsal midline of a rabbit, the fat pieces were grafted randomly into a pocket and the normal fat pieces into the other pocket as control. After 1, 3, 6, 9 and 12 months of implantation, the grafted fats were harvested, gross observation, weight measurement and histology were carried out. Number of the vessels stained with anti-CD34 antibody was counted out.

RESULTSVEGF concentrations in flaps were significantly higher (P < 0.05). The density of vessels in experimental groups increased significantly compared with that in control groups at 1 and 3 months, respectively (P < 0.01), and no significant differences in the survival rate of fat tissues between experimental and control groups were observed at 1 and 3 months (P > 0.05). The fat cells from the flaps survived after 12 months of fat plantation, while those in control groups disappeared after 6 months.

CONCLUSIONSThe survival rate and duration of grafted fat could be increased implanting the fat tissues from delayed fat flap, which may provide researchers with a new method for fat graft.

Adipocytes ; transplantation ; Adipose Tissue ; transplantation ; Animals ; Graft Survival ; Male ; Rabbits ; Surgical Flaps

OBJECTIVEBone-marrow derived mesenchymal stem cells (BMSC) have the potential to differentiate into endothelial cells. The aim of the study was to investigate the induction process of BMSC by B16 melanoma cells in vitro and to analyze the role of VEGF-a in the process.

METHODSA co-culture system containing BMSC and B16 melanoma cells based on transwell indirect model was established, and the induction process of BMSC by B16 melanoma cells was studied in vitro.

RESULTSBMSC were isolated from the bone marrow of C57 mice. BMSC expressed CD105, CD90, CD73, CD44 and CD166, and acquired expressin of endothelial phenotype markers including VEGFR-1, VEGFR-2 and Factor VIII after co-culture with B16 melanoma cells for 48 hours. The expression level of VEGFR-2 would be double and Factor VIII threefold more by extending the co-culture time to 72 hours. In the co-culture system, B16 melanoma cells also up-regulated the expression of VEGF-a.

CONCLUSIONSVEGF-a plays a significant role in the differentiation of BMSC into cells of endothelial phenotype, therefore, is important to tumor angiogenesis.

Animals ; Bone Marrow Cells ; cytology ; metabolism ; Cell Differentiation ; Cell Line, Tumor ; Coculture Techniques ; Endothelial Cells ; cytology ; metabolism ; Factor VIII ; metabolism ; Male ; Melanoma, Experimental ; metabolism ; pathology ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Mice ; Mice, Inbred C57BL ; Pilot Projects ; Vascular Endothelial Growth Factor A ; metabolism ; Vascular Endothelial Growth Factor Receptor-1 ; metabolism ; Vascular Endothelial Growth Factor Receptor-2 ; metabolism

BACKGROUNDThe mechanisms underlying postoperative pain remain unclear. Neurotransmitters of excitatory and inhibitory amino acids play an important role in the transmission and modulation of pain in the spinal dorsal horn. This study aimed to investigate the changes of release of excitatory and inhibitory amino acids in the spinal cord during postoperative pain and to provide a novel theoretical basis for postoperative pain management.

METHODSLoop microdialysis catheters were implanted subarachnoidally via the atlanto-occipital membrane in 16 healthy Sprague-Dawley rats. All rats without neural deficits were divided into two groups, Group A and Group B, following 5 days of recovery. The tubes for microdialysis were connected and 25 microl microdialysate sample for baseline value was collected after one-hour washout in each rat. A plantar incision in the right hind paws of rats in Group A were performed under 1.2% isoflurane. All rats in Group B were only anesthetized by 1.2% isoflurane for the same duration. The microdialysate samples were collected at 3 hours, 1 day, 2 days and 3 days after the incision (or isoflurane anesthesia in Group B) in both groups. The cumulative pain scores were also assessed at the above time-points. The amino acids in the microdialysate samples were tested using high performance liquid chromatography.

RESULTSWithin Group A, the release of aspartate and glutamate at 3 hours after the incision was significantly higher than the baseline values and the release of glycine at 1 day after the incision significantly increased compared with the baseline values (P < 0.01). Within Group B, the release of neurotransmitters at each time point had no significant difference compared with the baseline values (P > 0.05). The release of aspartate and glutamate at 3 hours after the incision in Group A was significantly higher than that in Group B (P < 0.01). The release of glycine at 1 day after the incision in Group A significantly increased compared with Group B (P < 0.01). The cumulative pain scores at 3 hours, 1 day and 2 days after the incision in Group A were significantly higher than those in Group B (P < 0.01).

CONCLUSIONSThe release of the excitatory amino acids occurs in the early phase of postoperative pain and might not be involved in the maintenance of pain in a rat model of incision pain. The release of inhibitory glycine lagged behind the excitatory amino acids. The implication of inhibitory glycine release remained to be established further.

Animals ; Aspartic Acid ; secretion ; Excitatory Amino Acids ; cerebrospinal fluid ; secretion ; Glutamic Acid ; secretion ; Glycine ; secretion ; Male ; Microdialysis ; Neurotransmitter Agents ; secretion ; Pain, Postoperative ; metabolism ; Rats ; Rats, Sprague-Dawley ; Spinal Cord ; secretion

OBJECTIVETo investigate the possibility to enhance the proliferation of adipose-derived stem cells (ASCs) in a delayed fat flap in rabbits.

METHODSA delayed fat flap was formed in one side of inguinal region of a rabbit. 21 days after operation, the fat tissues at the delayed flaps and at the unoperated side were harvested and digested with 0.25% collagenase and sieved. The cell suspensions were centrifuged. The cells were obtained from tissue precipitate after centrifugation. The expression rates of the surface marker (CD29, CD44, CD14 and CD45) were measured by FCM and compared between the experimental and control groups.

RESULTSExpression rates of CD29 and CD44 were higher in the delayed fat flap (74.06% and 90.74%) than in the contralateral fat tissue (62.88% and 77.54%, P < 0.05), while those of CD14 and CD45 were lower in the delayed fat flap (57.66% and 4.84%) than in the contralateral fat tissue (72.10% and 75.82%, P < 0.05 and P < 0.01).

CONCLUSIONSTissue hypoxic ischemia such as fat tissue in a delayed fat flap can promote proliferation of ASCs. It indicates that tissue in the delayed flap may be transplanted with better survival rate. The ischemia pretreatment of fat tissue may become a new method for fat transplantation.

Adipose Tissue ; cytology ; transplantation ; Animals ; Cell Proliferation ; Cells, Cultured ; Graft Survival ; Postoperative Period ; Rabbits ; Stem Cells ; cytology ; Surgical Flaps

Objective: To evaluate the efficacy of the third-generation instrumentation including TSRH, CD and ISOLA in the treatment of adult scoliosis. Methods:Thirty-five adult patients with idiopathic or degenerative scoliosis who received treatment with third-generation instrumentation (TSRH,CD and ISOLA) between July 1999 to January 2003 were retrospectively reviewed. The mean preoperative cobb angle of major curves of the frontal plane was 58.1°(42°-95°). The patients received a combined anteroposterior approach or a single posterior procedure. The mean follow-up time was 20 months(10-48 months). Preoperative and postoperative Cobb angles of the frontal plane and sagittal plane and the distance between C7 and CVLS were measured. The subjective assessment was judged by questionnaire. Results: Postoperative clinical appearance of all patients improved significantly. Mean correction of major curves of the coronal plane was 53.2%. Mean loss of correction of the coronal plane in the last follow-up was 4.3°. The distance between the midline of C7 and CVSL was corrected from 2.6 cm to 0.24 cm. The results of follow-up showed that 89.3% patients were satisfied with the outcome. Pneumatothorax and haematothorax occurred in 2 patients. Three patients still complained of low back pain one year after operation because of adjacent degeneration in 2 patients and pseudoarthrosis in the remaining 1 patient. Conclusion: Imageologic findings and subjective assessment of the patients showed that the third-generation instrumentation can achieve good correction and trunk balance in the treatment of adult scoliosis with fewer complications.

Objective: To evaluate the efficacy of the third-generation instrumentation including TSRH, CD and ISOLA in the treatment of adult scoliosis. Methods:Thirty-five adult patients with idiopathic or degenerative scoliosis who received treatment with third-generation instrumentation (TSRH,CD and ISOLA) between July 1999 to January 2003 were retrospectively reviewed. The mean preoperative cobb angle of major curves of the frontal plane was 58.1°(42°-95°). The patients received a combined anteroposterior approach or a single posterior procedure. The mean follow-up time was 20 months(10-48 months). Preoperative and postoperative Cobb angles of the frontal plane and sagittal plane and the distance between C7 and CVLS were measured. The subjective assessment was judged by questionnaire. Results: Postoperative clinical appearance of all patients improved significantly. Mean correction of major curves of the coronal plane was 53.2%. Mean loss of correction of the coronal plane in the last follow-up was 4.3°. The distance between the midline of C7 and CVSL was corrected from 2.6 cm to 0.24 cm. The results of follow-up showed that 89.3% patients were satisfied with the outcome. Pneumatothorax and haematothorax occurred in 2 patients. Three patients still complained of low back pain one year after operation because of adjacent degeneration in 2 patients and pseudoarthrosis in the remaining 1 patient. Conclusion: Imageologic findings and subjective assessment of the patients showed that the third-generation instrumentation can achieve good correction and trunk balance in the treatment of adult scoliosis with fewer complications.

BACKGROUNDAirway smooth muscle (ASM) is suspected to be a determining factor in the structural change of asthma. However, the role of protein kinase C alpha (PKCalpha) and cyclin D1 involved in the dysfunction of ASM leading to asthmatic symptoms is not clear. In this study, the central role of PKCalpha and cyclin D1 in ASM proliferation in asthmatic rats was explored.

METHODSThirty-six pathogen-free male Brown Norway (BN) rats were randomly divided into 2 groups: control groups (group N1, N2 and N3) and asthmatic groups (group A1, A2, and A3). Groups A1, A2 and A3 were challenged with ovalbumin (OA) for 2 weeks, 4 weeks and 8 weeks respectively. Control animals were exposed to an aerosolized sterile phosphate buffered saline (PBS). The ASM mass and nucleus numbers were studied to estimate the degree of airway remodeling by the hematoxylin-eosin staining method. PKCalpha and cyclin D1 expression in the ASM cells was detected by reverse transcription polymerase chain reaction (RT-PCR) and immunohistochemistry. The relation between PKCalpha and cyclin D1 was assessed by linear regression analysis. PKC agonist phorbol 12-myristate 13-acetate (PMA), PKC inhibitor Ro31-8220 and an antisense oligonucleotide against cyclin D1 (ASOND) were used to treat ASM cells (ASMCs) obtained from the 2 weeks asthmatic rats. The cyclin D1 protein expression level was detected by Western blotting.

RESULTSCompared with the control group, the PKCalpha and cyclin D1 mRNA levels were increased in the asthmatic group. Similar to RT-PCR results, immunohistochemistry analysis for PKCalpha and cyclin D1 expression revealed an increased production in ASMCs after allergen treatment for 2, 4 and 8 weeks compared with the respective control groups. No difference in expression of PKCalpha and cyclin D1 in ASM were found in the 2, 4 or 8 weeks asthmatic rats. There were significant positive correlations between PKCalpha and cyclin D1 expression, both transcriptionally (r = 0.944, P < 0.01) and translationally (r = 0.826, P < 0.01), in ASM. The content of cyclin D1 in asthmatic ASMCs increased after being stimulated by PMA, and decreased when induced by Ro31-8220. ASOND targeting for cyclin D1 lowered the expression of cyclin D1 induced by PMA.

CONCLUSIONSIncreased expression of PKCalpha and cyclin D1 in ASM along with smooth muscle structure changes might implicate PKCalpha and cyclin D1 participation in the proliferation of ASM and contribute to the pathogenesis of asthma after repeated allergen exposure in rats. The results suggested that cyclin D1 might be downstream of PKC signal transduction pathway.

Animals ; Asthma ; pathology ; Cell Proliferation ; Cyclin D1 ; genetics ; physiology ; Lung ; pathology ; Male ; Myocytes, Smooth Muscle ; pathology ; Protein Kinase C-alpha ; genetics ; physiology ; RNA, Messenger ; analysis ; Rats ; Rats, Inbred BN

This study is to investigate the expression of CyclinD1 in asthmatic rats and construct expression plasmids of sense and antisense CyclinD1 gene and transfect them to asthmatic airway smooth muscle cell to study the effects of CyclinD1 on the proliferation of airway smooth muscle cells in asthmatic rats. CyclinD1 cDNA was obtained by RT-PCR of total RNA extracted from the airway smooth muscle in asthmatic rats. The sequence was inserted into eukaryotic expression vector pcDNA3.1 (+) to recombinate the sense and antisense pcDNA3.1-CyclinD1 eukaryotic expression vector. The two recombinations and vector were then separately transfected into airway smooth muscle cell in asthmatic rats by using liposome. The expression level of CyclinD1 was certificated by Western blotting analysis. The proliferations of ASMCs isolated from asthmatic rats were examined with cell cycle analysis, MTT colorimetric assay and proliferating cell nuclear antigen (PCNA) immunocytochemical staining. Results showed (1) Compared with control group, the content of CyclinD1 was significantly increased; (2) It was comformed by restriction endonucleasa digestion and DNA sequence analysis that the expression plasmid of sense and antisense CyclinD1 were successfully recombinated. There was significant change of CyclinD1 expression between vector and sense CyclinD1 transfected cells, and the expression level of CyclinD1 in ASMC transfected with antisense CyclinD1 was lower than that in vector transfected cells (P <0.01); (3) In the asthmatic groups, compared with the vecter group, the percentage of S + G2M phase, absorbance A value of MTT and the expression rate of PCNA protein in ASMC transfected with pcDNA3. 1-CyclinD1 vector significantly increased. The values decreased remarkably in the pcDNA3,1-as CyclinD1 group. Statistical analysis revealed that there were significant differences in these indicators of cell proliferation in three groups (P <0.01). In the normal groups, statistical analysis revealed that there were significant differences in the percentage of S + G2M phase, a value of MTT and the expression rate of PCNA protein in three groups (P <0.01). Sense CyclinD1 eukaryotic expression vectors could have a positive effect on the proliferation of ASMC, however the antisence one have a negative effect, which implicated that CyclinD1 might contribute to the process of airway smooth muscle cell proliferation.
Animals ; Asthma ; pathology ; Cell Cycle ; drug effects ; Cell Proliferation ; drug effects ; Codon ; genetics ; pharmacology ; Cyclin D1 ; agonists ; antagonists & inhibitors ; genetics ; DNA, Antisense ; genetics ; pharmacology ; Disease Models, Animal ; Gene Expression ; Genetic Vectors ; genetics ; Male ; Myocytes, Smooth Muscle ; drug effects ; pathology ; Rats ; Rats, Sprague-Dawley ; Recombination, Genetic ; genetics ; Respiratory System ; Reverse Transcriptase Polymerase Chain Reaction ; Transduction, Genetic ; Transfection