Objective To study the regulatory effect of exogenous fibroblast growth factor(FGF\|2) in expressions of calcitonin gene\|related peptide(CGRP) and FGF\|2 in facial motoneurons after injury. Methods The right facial nerve was transected 6?mm distal to the stylomastoid foraman.A 3?mm\+3 piece of Gelfoam presoaked in 25??l of various test solutions(normal saline,or 16?10 6?IU/L FGF\|2,or 32?10 6?IU/L FGF\|2) was implanted adjacent to the proximal nerve stump.After three days survival,the rats were killed and brainstems were removed.Serial 20??m cryosections were cut through the whole brainstem.The CGRP and FGF\|2 immunoreactivity were studied by immunohistochemisty and image analysis. Results An increase in the CGRP and decrease in the FGF\|2 signals were presented in axotomized motoneurons with saline\|treated animals.In FGF\|2\|treated animals,the lesion\|induced up\|regulation of CGRP and down\|regulation of FGF\|2 were attenuated compairing with normal saline treated animals.Conclusion\ Exogenous FGF\|2 can increase FGF\|2 expression and decrease CGRP expression in rat facial motoneurons after injury.\;[

OBJECTIVE: To discuss the clinical characteristics and early diagnosis and treatment of relapsing polychondritis(RP). METHOD: Twenty-six RP patients received the therapies of antibiotics, glucocorticosteroid, immunosuppressive agent,tracheotomy, tracheal sent implantation or non-invasive ventilation. RESULT: Twenty-six cases (88.5%) had auricle involvement, 20 cases (76.9%) had respiratory tract involvement, 18 cases (69.2%) had joints involvement, 10 cases (38.5%) had nose involvement, 6 cases (23.1%) had eyes involvement, 4 cases (15.4%) had cochlea and (or) vestibular involvement. Some patients with cardiac, vascular, kidney damage as well. One case died of severe pulmonary infection. One case died of respiratory failure. The symptoms of other 24 cases were stable or improved in different degrees. CONCLUSION The clinical manifestations are diverse in RP patients, mainly in department of Otolaryngology, the prognoses of patients with airway involvement are worse, and it may improve the curative effect by an early diagnosis and a timely treatment.
Adult ; Aged ; Female ; Humans ; Male ; Middle Aged ; Polychondritis, Relapsing ; diagnosis ; therapy ; Prognosis ; Stents ; Tracheotomy ; Young Adult

The development of hearing of Kunming strain mouse was studied using evoked auditory nerve action potential (AP). The result showed that the onset of hearing was present during P10( tenth postnatal day) to P14. The thresholds of AP - N1 wave declined gradualy with the growth of the mouse. The auditory thresholds declined dramatically in the period from P10 to P20 and it indicated that the development of hearing was more rapid in this period. The thresholds, amplitudes and latencies of AP - N1 wave attained adult values by P40. The result showed that it took about one month for Kunming strain mouse to attain hearing maturation. The study on the development of hearing in normal mouse may serve as an important reference when studying degenerative lesions of the inner ear and the regeneration of cochlear hair cells.

BACKGROUND: The selection of carrier plays an essential role in the research of applying tissue-engineering to fix the peripheral nerves. An ideal carrier would be one that is similar to extracellular matrix and that it has biocompatibility with in vivo cells.OBJECTIVE: To evaluate the biocompatibility of hippocampal neural stem cells with collagen and gelatin sponge in vitro and to probe into the feasibility of using the materials as biomaterial scaffold in peripheral nerve tissue engineering.DESIGN: A randomized and controlled trial.SETTING: Department of Otorhinolaryngology of the First Affiliated Hospital of Zhengzhou University; Department of Anatomy of the School of Basic Medical Sciences of Zhengzhou University.MATERIALS: The experiment was carried out from July to December 2005 at the Laboratory of Neurobiology of the Department of Anatomy of the School College of Basic Medical Sciences of Zhengzhou University.Twelve New born (＜ 24 hours) clean grade guinea pigs of either gender with a body mass of 50-70 g, were provided by the Experimental Animals Center of Zhengzhou University School of Medicine.METHODS: The new born (＜ 24 hours) guinea pigs were anesthetized intraperitoneally with 10 g/L chloral hydrate and sterilized in 0.75 volume fraction of alcohol. Hippocampal tissue was resected from the brain under a surgical microscope. Hippocampus neural stem cells were cultured in vitro. The cultured cells of two generations were suspended at a density of 1 ×1010L-1 and respectively combined with collagen and gelatin sponge.The number of cells was counted and histological changes were observed under an inverted phase microscope and scanning electron microscope after 7 days, and the adhesion rate of the two materials to the cells were measured.MAIN OUTCOME MEASURES: ①Growth of the neural stem cells and their adhesion to the collagen and gelatin sponge were observed and the total number of the cells and adhesion rate with carrier were measured. ②The morphological changes of differentiation cells by immunocytochemical staining.RESULTS: ①Hippocampus neural stem cells could grew on the collagen and gelatin sponge and attached to them gradually. The adsorption rate of collagen was higher than thai of gelatin sponge (37.17 % and 14.87 %,x2=4.819,P ＜ 0.05). ② There was no significant difference in the total number of the cells in the control group, collagen group and gelatin sponge group [(53.17±3.5)×104,(53.25±2.6)×104, (52.04±4.05)×104,F=0.233,P ＞ 0.05].CONCLUSION: Two biomaterials (collagen and glutin sponge), especially collagen, have good biocompatibility with hippocampus neural stem cells from the guinea pigs and can be used safely as scaffold materials in peripheral nerve tissue engineering.

OBJECTIVE To evaluate the effectiveness of applying neuronal stem cells to bridging a 10mm long facial nerve defect in rabbits using a tissue engineered artificial nerve. METHODS Thirty-six Japanese white rabbits were randomly divided into three groups of 12. Group A: chitosan guidance channels with collagen protein sponge were filled with neuronal stem cells and NGF; Group B: chitosan guidance channels with collagen protein sponge were filled with NGF; Group C: autograft nerve. A series of examinations were performed, including electrophsiological methods, histological staining of nerve, Brdu and S100 immunohistochemical staining, at 12 weeks after operation. RESULTS At 12 weeks after operation, all examination results from Group A were better than from Group B(P0.05). CONCLUSION Neuronal stem cells can be used as seeding cells in peripheral nerve tissue engineering for repairing 10mm facial nerve defects.

AIM: To isolate and culture the adipose-derived stem cells (ADSCs) from guinea pig so as to create bases for the experimental study on treating sensorineural deafness by ADSCs transplantation. METHODS: The experiment was carried out between December 2005 and March 2006 in the Open and Key Laboratory of Clinical Medicine at Henan Universities. The inguinal fat pads were excised from adult male guinea pig weighing 500-750 g. A great quantity of adhesive and natant blood cells were observed under inserted microscope and phosphate buffer was used to wash natant cells repeatedly. Then the cells were cultured in DMEM with 10% fetal bovine serum. Before passage, the cells were washed by a small quantity of phosphate buffer and then cultured in 0.25% trypsin and 0.02% EDTA (2 mL). Then most of the ADSCs presented cytoplasm recovery and round shape, followed by digestion of 2 mL DMEM with feral bovine serum. Cellular suspension was collected to count the cells that were incubated at the density of 2?104/cm2. The morphology of ADSCs was observed constantly. Growth curve of ADSCs at passages 1, 3, 10 were recorded and detected by MTT. Surface markers of ADSCs were detected by flow cytometry. RESULTS: ①The result of primary culture showed that ADSCs at day 2 began to adhesion and more than 90 % of ADSCs at day 7 were approximately fusiform or fibroblast-shaped.②MTT detection showed that ADSCs had the capability to proliferate successively, especially at passage 1, 2, 3. But ADSCs at passage 10 presented decreasing proliferation. After digestion, the cells at passages 1, 3, 10 experienced the latent period (24-48 hours), logarithmic phase (3-5 days) and growth platform phase.③CD105 and CD44 were positive by flow cytometry, whereas CD34 was negative. CONCLUSION: Due to stable growth and rapid proliferation, the ADSCs from guinea pig can be used as the donor cells in the treatment of sensorineural deafness by stem cell transplantation.

Objective To study the inhibitory effect and its mechanism of sodium butyrate on human laryngeal carcinoma in nude mice. Methods Human laryngeal carcinoma cell line Hep-2 was seeded in the subcutaneous layer of 12 nude mice to built laryngeal carcinoma xenograft model. Then they were randomly and equally divided into 2 groups. Sodium butyrate was given in experimental group while phosphatic-buffered saline (PBS) was used in control group for 4 weeks. Tumor size and body weight of the mice were measured at regular time-intervals. The tumor,heart,liver,lungs,spleen and kidneys were removed at the end of treatment. Tumor sections were examined by electronic microscopy. TUNEL method and immunohistochemical S-P method were used for detecting the expression of Ki-67 nuclear antigen and survivin protein. The heart,liver,lung,spleen and kidney sections were examined after HE staining for assessment of toxicity. Results In experimental group,the volume of tumors was reduced,the area of necrosis in tumors was widened,the apoptotic rate was increased obviously and the expression level of Ki-67 nuclear antigen and survivin protein was decreased as compared with control group. During treatment,all the nude mice grew well and there were no toxic reactions. At the end of treatment,there were no abnormal changes in heart,liver,lung,spleen and kidney sections examined under light microscope. Conclusion Sodium butyrate can significantly inhibit the growth of human laryngeal carcinoma xenograft in nude mice. Its mechanism may be related to the apoptosis in tumor cells by inhibiting the expression of survivin protein and Ki-67 nuclear antigen. There is no toxicity to heart,liver,lungs,spleen and kidneys at a treatment dose of sodium butyrate.

BACKGROUND: Transplantation of neural stem cells into inner ear brings hope for treatment of sensorineural deafness, while guinea pig is the preferred animal in developing animal models of aminoglycosides poisoning-induced deafness. Successful animal models have been established after study for several scores of years. How to isolate neural stem cells and label them from hippocampi of guinea pigs is one of the problems to be solved in transplantation of stem cells into inner ear.OBJECTIVE: To isolate and culture neural stem cells from hippocampi of newborn guinea pigs so as to lay basis for transplantation of neural stem cell in the treatment of sensorineural deafness.DESIGN: Single sample observation SETTING: Staff Room of Microbiology and Immunology, Zhengzhou University MATERIALS: This experiment was carried out at the Staff Room of Microbiology and Immunology, Zhengzhou University from March to June 2005. Newbornguinea pigs born within 24 hours of either gender, weighing 50 to 80 g, provided by Experimental Animal Center of Zhengzhou University were used in this experiment.METHODS: Neural stem cells from the hippocampal tissue of newborn guinea pigs were isolated and cultured by serum-free medium containing basic fibroblast growth factor (bFGF) and epidermal growth factor (EGF).Expression of nidogen was detected with immunocytochemical staining.Neural stem cells were labeled by DAPI.MAIN OUTCOME MEASURES: ① In vitro growth of neural stem cells; ② neural stem cells nidogen expression was identified by immunofluorescent staining; ③ labeled stem cells by fluorescence and detected the labeling rate.RESULTS: ①Clone-like cluster of neural stem cells were obtained from hippocampal tissue ② Neural stem cells could express nidogen. ③Labeling rate of DAPI was 93.4% and no attenuation of fluorescence brightness as observed after 8 generations in culture.CONCLUSION: The cells obtained from hippocampal tissues of newborn guinea pigs can be used as donor cells in the treatment of neurosensory deafness by transplanting neural stem cells, for their obvious proliferation ability and high-labeled efficiency with fluorescent dye.

Objective: To discuss how to improve the level of diagnosis and treatment about the firstbranchial fistula. Method:16 cases with the first branchial fistula were analyzed retrospectively. Result :Of all thepatients underwent surgery, 14 cases were free from disease postoperatively within 0.5 years follow-up, 2patients underwent re-operation because of recurrence. Conclusion:Knowing about the disease fully, selectingproper surgical incision and possessing skilled surgical technique is important to improve the level of diagnosisand treatment about the first branchial fistula.

Objective:To investigate the clinicopathlogical significance of the CyclinE,p21WAF1/CIP1 and p53 expression in laryngeal carcinogenesis.Method:The expression of CyclinE,p21WAF1/CIP1 and p53 was examined by using immunohistochemical method in normal epithlia (20),dysplastic epithlia (40) and laryngeal cancers (60).Result:①In normal epithlium,dysplastic squamous epithlium and in squamous cell carcinoma of the larynx,the positive immunostaining rate of CyclinE protein was 5.0%(1/20),20.0%(8/40) and 45.0%(27/60) respectively (P＜0.001),and p21WAF1/CIP1 protein was 95.0%(19/20)，75.0%(30/40) and 63.3%(38/60) respectively (P＜0.01),while the rate of p53 was 0,30.0%(12/40) and 61.7%(37/60)(P＜0.001) respectively.②p21WAF1/CIP1 positive expression in highly、moderately and poorly differtiated carcinomas were 76.2%(16/21)、66.5%(19/29) and 30.0%(3/10) respectively (P＜0.05),p21WAF1/CIP1 protein expression was significantly associated with tumour cell differentiation.③The positive expression of CyclinE closely related with the positive expression of p53 (P＜0.05).No correlation was found between p21WAF1/CIP1 expression and p53 expression.Conclusion:①CyclinE and p53 could be an early molecular event in the carcinogenesis of larynx.②p21WAF1/CIP1 expression is associated with tumour cell differentiation.③The mutual mechenism of cell cycle regulators (CyclinE、p21WAF1/CIP1 and p53) play an important role in the laryngeal carcinoma.