Humans ; Tinnitus ; diagnosis ; therapy

Cartilage ; transplantation ; Ear, Middle ; surgery ; Humans ; Otologic Surgical Procedures ; methods

Objective　To obtain staphylococcal enterotoxin B (SEB) mutant with normal antigenicity but low toxicity. Methods　Using PCR technique, normal SEB (SEB-N) gene which was amplified from S. aureus S6B. SEB mutant gene (SEB-M) was prepared from the same strain, but one nucleotide in SEB gene was changed from asparagine (N23) to serine (S23). SEB-N and SEB-M were cloned into procaryotic expression vector pTrc99A then and transferred into E. coli JM109. SEB-N and SEB-M which were cloned into plasmid were sequenced directly by dideoxynucleotide method. The crude expressed proteins were identified by double agar immunodiffusion. The level of IL-2 in supernatants of mouse splenocytes stimulated by crude expressed proteins was determined by ELISA. Results　SEB-N and SEB-M were obtained through PCR. The sequence of SEB-N was changed with non site-directed mutagenesis, threonine at the residue 150 of SEB-N was replaced with alanine (ACT→GCT, T150A). As being expected, at the residue 23 of SEB-M, serine substituted for asparagine (AAT→AGT, N23S) with site-directed mutagenesis. Double agar immunodiffusion showed obvious precipitin line with anti-SEB by both crude SEB-N and SEB-M mutant proteins could produce, but not by non-recombinant strain. ELISA demonstrated that the level of IL-2 in supernatant of mouse splenocytes stimulated by natural SEB protein (containing equal amount of JM109P crude protein) was 40 times as much as that stimulated by SEB-M and 12.5 times as much as that stimulated by SEB-N. Conclusions　We obtained two recombinant strains which produced T150A and N23S mutant SEB protein. The mutant proteins showed binding ability to anti-SEB as the normal protein. However, their biological activity as superantigen decreased sharply. We consider that it is promising for further study of molecular adjuvant or superantigen vaccine.

OBJECTIVE To measure the dexamethasone concentration in the cochlear tissue after postaurical and intramuscular administration. METHODS After bilateral adrenalectomy,animals were divided into three groups:postaurical injection group,intramuscular injection group,and physiological saline injected blank control group.At 0.25,0.5, 2.5,5,12,24,72 h after injection(1mg/100g), cochleae together with the intact endolymphatic sac were removed and homogenated.After that, the dexamethasone was detected by enzymelinked immunosorbent assay(ELISA).RESULTS The dexamethasone levels in the cochlear tissue in postaurical injection group were significantly higher than that in the intramuscular injection group.In the postaurical administration group,a mean peak cochlear tissue concentration of(175.2?36.0) ng/ml was detected at 0.25 h after injection,then declined obviously 12 h and was below detection limits by 24h.While in the intramuscular administration group, a mean peak concentration was observed 2.5 h at the concentration of(141.1?8.9) ng/ml,then rapidly declined from 5h and beyond detection by 24 h.CONCLUSION Compared with intramuscular administration,the dexamethasone levels in the postaurical injection group were significantly higher, with peak concentration reached earlier,and last longer.

Objective To explore the common influencing factors on the severity of Reinke's edema of the vo-cal cord and to analyze the factors of recurrence.Methods To analyze the relationships among the degrees and smoking,voice overuse,and laryngopharyngeal reflux for 85 patients with Reinke's edema of the vocal cord,and an-alyze the risk factors of recurrence.ResuIts 84.7% of the patients were male with the mean age of 51.35 years. There were statistical differences among smoking history of more than 25 years (P<0.001),laryngopharyngeal re-flux (P<0.01),and voice overuse (P<0.01).Follow ups lasted 1 year and 7 patients showed recurrence.ConcIu-sion Reinke's edema was related to long - term smoking and common in senior male patients.The degree of Reinke's edema was positive correlation with smoking,laryngopharyngeal reflux and voice overuse.The most impor-tant factor is smoking history of more than 25 years.The longer smoking history,the more severe Reinke's edema.

B and T lymphocyte attenuator (BTLA) is an immune checkpoint discovered in the early 21st century. BTLA is a typeⅠtransmembrane protein that belongs to Ig super family. The ligand for BTLA is herpesvirus-entry mediator (HVEM), a TNF receptor family protein. BTLA regulates the homeostasis of γδ T cells and innate lymphocytes (ILCs), promotes the survival of effector T cells and helps them differentiate into memory T cells. BTLA also plays an important role in maintaining the homeostasis of dendritic cells (DCs). Studies have shown that BTLA knockout mice have increased susceptibility to autoimmune diseases. Recent studies showed that BTLA also plays a crucial role in allergic diseases (such as allergic conjunctivitis and allergic rhinitis) and autoimmune diseases (such as systemic lupus erythematosus and ankylosing spondylitis). This article makes a systematic review about recent researches of BTLA in immune diseases.

Objective To study the correlation between dopamine and glutamate receptor NMDA NR_1 and NMDA NR_(2A),and to share the understanding of the mechanism of dopamine in the synaptic complex of inner hair cells.Methods Forty guinea pigs were divided randomly into four groups and the whole intacochlear perfusions were performed.The perfused cochleas were taken out as preparations 2 hours after perfusing,the contralateral cochleas were also taken out as the normal control group in the group perfused with artifical perilymph solutions.All the preparations were divided into 5 groups:①normal control cochleas;②perfused with artificial perilymph solutions;③perfused with artifical perilymph solutions containing 10 mmol/L dopamine；④perfused with artificial perilymph so lutions containing 30 mmol/L dopamine；⑤perfused with artifical perilymph solutions containing 50 mmol/L dopa mine.The semi-quantitive RT-PCR was used to observe the difference in the amount of glutamate receptor NMDA NR_1、NMDA NR_(2A).Results Dopamine inhibited the compound action potential(CAP),the increase of CAP threshold was observed and correlated with the contentration of dopamine in the perfusion solution.Regarding the amount of glutamate receptor NMDA NR_1 mRNA,there was no significant difference between group ① and group ② (P＞0.05).But a significant difference was observed the other 3 groups when compared to group ①(P＜0.05).No significant difference was detected among the 5 groups in the amount of glutamate receptor NMDA NR_(2A) (P＞0.05).Conclusion Dopamine may inhibit the cochlear auditory afferent nerve.The significant correlation between dopamine and glutamate receptor NMDA NR_1 was observed,the amount of glutamate receptor NMDA NR_1 decreased along with the increasing of the contentration of dopamine in the perfusion solution.And there was no significant correlation between dopamine and glutamate receptor NMDA NR_(2A).

Disease Management ; Humans ; Tinnitus ; therapy

OBJECTIVE To investigate the protective effect and its frequency selection of dopamine on the inner cell in cochlea when exposed to white-noise;In order to offer an important step on understanding of negative protective modulation of dopamine in the cochlear auditory afferent nerve. METHODS Forty guinea pigs were randomly divided into 4 groups and whole intracochlear perfusions were performed. ① exposed to the 100dB white-noise; ② perfused with artificial perilymph solutions; ③exposed to the white-noise and perfused with artificial perilymph solutions; ④exposed to the white-noise and perfused with Artificial perilymph solutions containing 1mmol/L dopamine. Compound action potential(CAP)evoked by different frequencies tone pip, and cochlear mirophonics(CM)evoked by 4kHz tone burst were recorded from the round window of guinea pigs before and 2h after perfusion. RESULTS In the groups exposed to the white-noise, the CM amplitude and the linearity of input-output function both significantly decreased at 2h(P

OBJECTIVE To analyze 1000 Hz probe tone tympanometry of middle ear with different volume of fluid, and to evaluate the role of 1000 Hz probe tone tympanometry in the diagnosis of presence of fluid in tympanic cavity. METHODS Tympanometries with 1000 Hz or 226 Hz probe tone were obtained from all the guinea pigs with different volume of fluid in tympanic cavity using GSI33 analyzer and tympanometry curves were analyzed. RESULTS There was a significant difference of the peak pressure of 1000 Hz tympanometry among all test groups and control group. The peak pressure decreased with the increasing of fluid volume in tympanic cavity. CONCLUSION Tympanometry with 1000 Hz probe tone can be used in the diagnosis of fluid in tympanic cavity, and it can be an indicator of volume of fluid in tympanic cavity.