Objective　To set up a reliable method for isolating and culturing cochlea microvascular endothelial cells. Methods　Cochlea stria was separated by micrergy from guinea pigs and the stria tissue nubbles were cultured in vitro. Results　Two days later, a few cells were seen disseminately around partial tissue nubbles and their number was increased with time prolongation. There were some cellular colonies consisted of large quantity of cells around the partial tissue nubbles on 10 th day. Under the inversion microscope, the single cultured cells presented a long fusiform and the confluent monolayer cells linked tightly and exhibited the typical structure of “scree” of cultured endothelial cells in vitro. After purification, over 95% of the cultured cells showed factorⅧ relative antigen positive reaction. Conclusion　The method used in this study can obtain cochlea microvascalar endothelial cells of guinea pig in vitro.

Objective To set up reliable cell culture methods of cochlear microvascular endothelial cells in guinea pigs.Methods The cochlear striae vascularies were acquired from guinea pigs with micrergy, the cells were cultured and pured by tissue nubbles cultivation and collagenase digestion.Results Two days after tissue nubbles cultivation of striae vascularis, cells grew at the edge of some tissue nubbles,and proliferated to massive colonies composed of more cells;Some islands made up of a few cells on the bottom of Petri dish were found one to three days after striae vascularies tissue was digested by collagenase;These cells were mainly polygon after purification, monolayer confluent culture cells showed typical "flagstone" appearance as cul tured endothelial cells. According to immunohistochemical detection of factor VIII, which is the mark antigen of endothelial cell, we had observed that the positive reaction cells with cytoplasmic brown pigment were over 95%.Conclusion Cochlear microvascular endothelial cells from guinea pigs can be acquired by tissue nubbles cultivation and collagenase digestion .

Objective To evaluate endoscopic minimally invasive surgery in the treatment of sphenoidal and ethmoidical mucoceles. Methods A total of 25 patients diagnosed as sinus mucoceles, 10 in the sphenoid sinus and 15 in the ethmoid sinus, were subjected in this study, who underwent endoscopic marsupialization in our department from 1998 to 2000. Postoperatively, the patients were followed from 1 to 3 years. Results All 25 sphenoidal and ethmoidical mucoceles were satisfactorily approached endoscopically, and operating cavities received satisfactory marsupialization. The symptoms were improved or relieved in 23 patients. None had significant complications. Conclusion Endoscopic minimally invasive surgery is a safe and valuable way in the treatment of sphenoidical and ethmoidical mucoceles.

The author proposed student-centered learning system in otolaryngology-head and neck surgery for undergraduate clinical training after exploration and intentions.Four mutual impacted frames were built including integration of teaching philosophy,visualization of training methods,diversification of educational targets and interaction of training courses.Endoscopic navigated learning and multimedia aided training were applied,respective teaching purposes were set and various clinical training courses were introduced to students in their learning of otolaryngology,which were believed to help develop more medical talents with higher comprehensive qualities and better clinical skills.

Objective To study the initial application of 3.0T high field intensity intraoperative magnetic resonance(iMR)for cranial tumors.Methods Forty-three patients with cranial tumors including 23 glioma cases,12 pituitary tumor cases,3 brain stem cavernous hemangioma cases,2 meningioma cases,2metastatic tumor cases,1 neurilemmoma case,received operation examined with GE Signa HDX 3.0T iMR system.The operation process and influence of iMR were reviewed.Results In 43 patients,average iMR examination was 1.3(1-3)times.In 16 patients the first iMR examination revealed tumor remnants,and in 13 of them continued surgical interventions and complete resection.The rate of complete resection was increased from 63％(27/43)to 93％(40/43).No complications related to iMR occurred.Conclusion 3.0T high field intensity iMR can provide accurate positioning and real-time navigation for the surgery,increase the rate of complete resection,improve the accuracy and safety of cranial tumor resection,and decrease complications.

BACKGROUND: Vascular restenosis alter carotid endarteractomy (CEA) is an important factor affecting curative affect ofoperation.OBJECTIVE: To explore the role of matrix metalloproteinase-9 (MMP-9) mRNA dynamic expression in the development of early vascular restenosis after carotid endarterectomy.DESIGN, TIME AND SETTING: A random grouping contrast observation was completed in the General Hospital of Beijing Military Area Command of Chinese PLA from February 2006 to December 2007. MATERIALS: Forty-one healthy male New Zealand rabbit, weighing about 3.0 kg, with 36 ones used for preparing carotid atherosclerotic stenosis (CASS) models. experimental group, each 6 of the CASS rabbit models (n =36) were selected at the time points of hour 4, day 1, 3, 7, 30, and 90 following CEA respectively. Then they were fixed with 40 g/L polyoxymethylene perfusion and stained with hematoxylin-eosin to observe their morphologic changes.MAIN OUTCOME MEASURES: The expression changes of MMP-9 mRNA were observed dudng the development of early vascular restenosis by the quantitative real-time polymerase chain reaction technique preoperatively as well as at day 1, 3 and 7 following CEA.RESULTS: Several stages could be seen in the reparative process of neointima after CEA, including the thrombosis, the inflammatory reaction, the repair of endothelium, the proliferation of vascular smooth muscle call, the formation and accumulation of extracellular matrix. MMP-9 mRNA was expressed since day 1, reached a peak at day 3 and then decreased significantly at day 7 postoperatively.CONCLUSION: MMP-9 plays an important role in the proliferation, migration and reconstruction of vascular smooth muscle calls, the mediated reconstruction of local blood vessels, as well as the development of vascular restenosis.

Through introducing mutations into ribosomes by obtaining spontaneous drug resistance of microorganisms, ribosome engineering technology is an effective approach to develop mutant strains that overproduce secondary metabolites. In this study, ribosome engineering was used to improve the yield of butenyl-spinosyns produced by Saccharopolyspora pogona by screening streptomycin resistant mutants. The yields of butenyl-spinosyns were then analyzed and compared with the parent strain. Among the mutants, S13 displayed the greatest increase in the yield of butenyl-spinosyns, which was 1.79 fold higher than that in the parent strain. Further analysis of the metabolite profile of S13 by mass spectrometry lead to the discovery of Spinosyn α1, which was absent from the parent strain. DNA sequencing showed that there existed two point mutations in the conserved regions of rpsL gene which encodes ribosomal protein S12 in S13. The mutations occurred a C to A and a C to T transversion mutations occurred at nucleotide pair 314 and 320 respectively, which resulted in the mutations of Proline (105) to Gultamine and Alanine (107) to Valine. It also demonstrated that S13 exhibited genetic stability even after five passages.
Genetic Engineering ; Macrolides ; metabolism ; Point Mutation ; Ribosomal Proteins ; genetics ; Ribosomes ; metabolism ; Saccharopolyspora ; metabolism

Objective:With the development of modern skull base minimally invasive technology mature and neural radio surgery techniques,it is necessary to re-examine the therapeutic strategy for the treat-ment of petroclival meningiomas.To sum up the operative experience and methods in microsurgical resec-tion of petroclival meningiomas by the combining trans-subtemporal and suboccipital retrosigmoid keyhole approach.To explore the minimally invasive operation approach of petroclival meningiomas,to raise the removal degree and to improve the postoperative result using this approach.Methods:The clinical data of the consecutive 21 patients with the petroclival meningiomas were reviewed retrospectively.The meth-od,degree of tumor resection,techniques of the combining keyhole approach,Karnofsky performance score (KPS)before and after operation were also analyzed.The neuronavigation guided operation was performed in 9 cases,and 12 cases were operated in the neuroelectrophysiological monitoring.Results:Total excision of the tumor resection (Simpson,Ⅰ -Ⅱlevels)was conducted in 18 cases (85.7%,18 /21),and 3 patients underwent close resection (Simpson Ⅲ level,14.3%,3 /21).Postoperative three-dimensional CT showed good lock bone flap restoration;Postoperative pathology confirmed meningioma. Postoperative cranial nerve dysfunction or new original nerve dysfunction were aggravated in 5 cases (23.8%),including transient trochlear nerve (3 cases),abducent nerve (1 case),and the motor branch of trigeminal nerve paralysis (1 case).Abducent nerve paralysis (1 case)appeared,with hearing impairment.After the 3-month follow-up,11 cases had the same KPS aspreoperation,7 cases improved,and 3 cases not improved.The KPS score was 77.14 ±23.12 on average,and there was no statistically significant difference compared with that before operation (P >0.05 ).The postoperative follow-up for half a year showed fluent speaking and writing in 19 cases (KPS 70 or higher),and general recovery in 2 cases (KPS <70).The postoperative follow-up for 3 -29 months showed no tumor recur-rence or progress.Conclusion:The combining trans-subtemporal and suboccipital retrosigmoid keyhole approach is simple,safe,and minimally invasive,and an ideal operation approach of petroclival menin-gioma.To master the operation skills and the intraoperative matters needing attention in the operation,is favorable to improve the resection rate and curative effect.

OBJECTIVETo construct a tissue-engineered skin flap using composite skin and adipose tissue constructed by adipose-derived stem cells(ASCs).

METHODSHuman ASCs isolated from adipose tissue were cultured and identified for their adipogenic, osteogenic and chondrogenic differentiation potentials. ASCs were then mixed with collagen gel for adipogenic induction and observed 15 days later with inverted microscope, oil-red O staining and HE staining. To construct the composite skin, keratinocytes and fibroblasts were isolated from human foreskin. The fibroblasts were mixed with collagen gel and cultured for 5 days, and keratinocytes were seeded on the gel for 4 days before transfer of the culture to air-liquid interface for culture for another 10 days. The adipose tissue and composite skin were then assembled according to the structure of normal skin and cultured for 3 days with HE staining observation.

RESULTSThe cultured ASCs were capable of adipogenic, osteogenic and chondrogenic differentiation, and adipogenic induction of the ASCs-gel complex for 15 days resulted in adipogenic differentiation of the ASCs in gel. The assembled tissue-engineered skin consisted of 3 layers, including a suprabasal layer formed by the stratified and differentiated keratinocytes, the middle layer and sublayer containing numerous cells, and a underlying sublayer formed by the adipogenic ASCs.

CONCLUSIONTissue-engineered skin flap can be constructed by assembling composite skin and adipose derived from cultured keratinocytes, fibroblasts, and ASCs.

Adipocytes ; cytology ; Adipose Tissue ; cytology ; Cell Culture Techniques ; Cells, Cultured ; Fibroblasts ; cytology ; Humans ; Keratinocytes ; cytology ; Stem Cells ; cytology ; Surgical Flaps ; Tissue Engineering

Objective: To study the epidemiological characteristics and related factors of dyslipidemia among adult residents in Xinjiang Uygur Autonomous Region (Xinjiang) in 2013-2014. Methods: A total of 4 120 adult residents consisting of Han and Uygur group aged over 18 years old were selected by using a stratified cluster random sampling method in 8 counties of Xinjiang from 2013 to 2014. The related factors of dyslipidemia were collected by questionnaire and physical measurement. The total cholesterol, triglyceride, high-density lipoprotein cholesterol and low-density lipoprotein cholesterol were detected by enzyme method. Factors associated with dyslipidemia were analyzed by chi-squared test and a multivariate unconditioned logistic regression model adjusted for gender, urban or rural area, age-group, body mass index (BMI), central obesity, smoking, drinking, education attainment, diabetes mellitus and hypertension. Results: The prevalence of dyslipidemia was 45.00% (1 854 cases). The prevalence of dyslipidemia was higher in Uygur group (47.80% (977/2 044)) than that in Han group (42.24% (877/2 076)) (χ²=12.84, P<0.001). The analysis showed that dyslipidemia was related with gender (OR=0.41, 95%CI: 0.33-0.51), urban area (OR=0.54, 95%CI: 0.39-0.76), BMI (overweight group (OR=1.52, 95%CI: 1.18-1.96); obesity group (OR=2.20, 95%CI: 1.64-2.96)), central obesity (OR=1.66, 95%CI: 1.29-2.14) and diabetes mellitus (OR=1.49, 95%CI: 1.06-2.11) in Uygur group. The analysis also showed that dyslipidemia was related with BMI (overweight group (OR=1.72, 95%CI: 1.32-2.25), obesity group (OR=2.60, 95%CI: 1.85-3.64)), central obesity (OR=1.45, 95%CI: 1.13-1.87), smoking (OR=1.46, 95%CI: 1.09-1.95), diabetes mellitus (OR=1.77, 95%CI: 1.38-2.25) and hypertension (OR=1.62, 95%CI: 1.31-2.00) in Han group. Conclusions The prevalence of dyslipidemia in Xinjiang was higher than the national average prevalence. The prevalence of dyslipidemia in Uygur group was significantly higher than that in Han group. The gender, living area, BMI, central obesity and diabetes mellitus were risk factors of dyslipidemia in Uygur group, and BMI, central obesity, smoking, diabetes mellitus and hypertension were risk factors of dyslipidemia in Han group in Xinjiang.