Frequency characteristics of horizontal semicircular canals damage and the ultrastructure analysis of crista ampullaris in patients with Meniere's disease / 中华耳鼻咽喉头颈外科杂志

Objective: To investigate the frequency characteristics and the pathological characteristics of the horizontal crista ampullaris in patients with Meniere's disease,and to analyse its structural basis. Methods: Between March, 2019 and November, 2019, seventy-two patients diagnosed as Meniere's disease (27 males and 45 females, aged from 13 to 74 years, with a course of disease ranging from 4 months to 32 years)in Shandong Provincial ENT Hospital were included.Caloric test, sinusoidal harmonic acceleration test (SHA), video-head impulse test (v-HIT), Gadolinium-enhanced inner-ear 3D-FLAIR MRI and pure tone audiometry were conducted in the patients. The function of the horizontal semicircular canal in these patients were analysed as well as its relationship with the degree of endolymphatic hydrops,clinical stage and duration. Light microscopy and transmission electron microscopy were used to observe the ultrastructure of horizontal semicircular canal crista ampullaris from six patients with refractory Meniere's disease who underwent labyrinthectomy. The number of type Ⅰ and type Ⅱ vestibular hair cells, the common pathophysiological changes of horizontal semicircular canal crista ampullaris were investigated in these patients. Statistical analysis was performed using SPSS 19.0. Results: With the increase of detection frequency, the abnormal rate decreased gradually. The abnormal rate of caloric test was 69.4% (50/72), SHA 51.4% (37/72), V-HIT 36.1% (26/72), comparation of the positive rate among the three tests showed statistically significant differences(P<0.05).Neither caloric test nor SHA had correlation with the degree of hydrops(P>0.05), but v-HIT(r=0.434,P<0.01).There was correlation with clinical stage to SHA and v-HIT(r=0.338,0.462,P<0.01), except caloric test(P>0.05).No significant relation was found with caloric test, SHA, v-HIT and course of disease(P>0.05).Morphological observation found abnormal monolayer epithelialization of the horizontal semicircular canal crista ampullaris significantly decreased number of type Ⅱ hair cells compared with type Ⅰhair cells. Hair cells showed perinuclear vacuolization, cytoplasmic vacuoles, mitochondrial electron density increasement and loss of stereocilia. Conclusions: The horizontal semicircular canal damage in the patients with Meniere's disease has a frequency-dependent characteristic, mainly occurres in low frequency area. With progress of the disease, the high frequency area of ampulla will be impaired gradually, and it is related to the degree of endolymphatic hydrops and hearing level. Hair cell injury would be observed,the frequency characteristics may be more associated with the disorder of type Ⅱ hair cells.

The Protective Effect of Epigallocatechin-3-Gallate Against Gentamicin Vestibular Ototoxicity in Type I Vestibular Hair Cell of Guinea Pig / 대한이비인후과학회지

BACKGROUND AND OBJECTIVES: Gentamicin (GM) is well known for its vestibulotoxicity. There have been many reports about vestibulotoxicity, however, its mechanism is still unclear. So far, it is known that GM affects the voltage-dependent K+ current and nitric oxide (NO) production. Epigallocatechin-3-gallate (EGCG) is the major component of green tea and is known to have anti-oxidative and anti-toxic effect. This study was undertaken to investigate the protective effect of EGCG against gentamicin on vestibular hair cell (VHC). MATERIALS AND METHOD: White guinea pigs (200-250 g) were rapidly decapitated and the temporal bones were immediately removed. Under a dissecting microscope, the crista ampullaris was obtained. The dissociated VHCs were transferred into a recording chamber mounted onto an inverted microscope. Whole-cell membrane currents and potentials were recorded using standard patch-clamp techniques. In addition, measurements of NO production were obtained using the NO-sensitive dye, 4,5-diamino-fluorescein diacetate (DAF-2DA). RESULTS: Type I VHCs Voltage-dependent K+ current was activated from low depolarizing stimulation. As the stimulation increased, higher current was detected. Voltage-dependent K+ current in type I VHCs was decreased when GM (200 microM) was administrated and GM effects of K+ current inhibition was significantly blocked by EGCG. Extracellular GM-induced an increase in DAF-2DA fluorescence, which thus indicates NO production in VHCs. Also, the GMinduced NO production was inhibited by EGCG. CONCLUSION: GM inhibits voltage-dependent K+ current by releasing NO in isolated type I VHCs. EGCG blocks this inhibitory effects, suggesting a protective role on GM vestibulotoxicity.

Investigation of the reverse phase nystagmus in positioning test for benign paroxysmal positional vertigo / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To investigate the clinical significances and formation mechanism by analyzing the characteristics of the reverse phase nystagmus parameters from benign paroxysmal positional vertigo (BPPV) positioning test.</p><p><b>METHODS</b>There were 183 cases with BPPV, including 108 cases (59.0%) of posterior semicircular canal canalithiasis, 55 cases (30.1%) of horizontal semicircular canal canalithiasis, and 15 cases (8.2%) of horizontal and posterior semicircular canal cupulolithiasis and 5 cases (2.7%) of anterior semicircular canalithiasis . The video-nystagmography was utilized in positioning tests to induce nystagmus. The direction, intensity, time parameters characteristic of vertical nystagmus in Dix-Hallpike test and horizontal nystagmus in roll test were analyzed and compared.</p><p><b>RESULTS</b>There were no reversal phase nystagmus in 15 cases of semicircular canal cupulolithiasis and 5 cases of anterior semicircular canalithiasis. After the disappearance of vertical nystagmus which induced by hanging position (the first phase nystagmus) in 108 cases of posterior semicircular canalithiasis of Dix-Hallpike test, there was 54 cases(50.0%) of posterior semicircular canal canalithiasis displayed downward vertical nystagmus (reverse phase nystagmus) . The latency, duration time and intensity of the first phase nystagmus and reverse phase nystagmus were [(2.00 ± 1.10) s, (3.54 ± 1.42) s], [ (16.27 ± 4.95) s, (61.65 ± 33.69)s] and [ (51.80 ± 25.25) °/s, (10.65 ± 6.29)°/s] respectively; 43 cases(78.2%) of horizontal semicircular canal canalithiasis displayed the opposite to turning head (reverse phase nystagmus) after the horizontal nystagmus, similar with turning head disappeared in Roll test. The latency, duration time and intensity of the first phase of nystagmus and reverse phase nystagmus were [ (1.67 ± 1.07) s, (3.57 ± 1.89)s], [ (25.19 ± 9.74) s, (70.48 ± 40.26)s] and [ (68.47 ± 30.18) °/s, (11.22 ± 8.78)°/s] respectively. Comparing with the latency, duration time, intensity of first phase nystagmus and reverse phase nystagmus of posterior and horizontal semicircular canal canalithiasis, the differences had statistical significances (P < 0.05). Comparing with the first phase nystagmus of reverse phase and no reverse phase nystagmus canalithiasis, the difference of nystagmus intensity had statistical significances (P < 0.05); but the differences of latency and duration of nystagmus had no significant difference (P > 0.05).</p><p><b>CONCLUSIONS</b>It is common in PSC-Can and HSC-Can patients that reverse phase nystagmus is one of the clinical features of canalithiasis. It appears in side head position of Rolling test or the hanging of Dix- Hallpike test. More power of the first phase nystagmus has, reverse phase nystagmus will be induced much easier. In comparison of the reverse phase nystagmus, the first phase nystagmus has the shorter incubation and duration, but it has more power. It is helpful to avoid interruptions of clinical statolith positioning and reset since we recognize the reverse phase nystagmus. The mechanism might be similar to the vestibular mirror image nystagmus. It is another form of the vestibular mirror image nystagmus in BPPV patients.</p>

Analysis of the dysfunction frequency and characteristics of semicircular canal in benign paroxysmal positional vertigo / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To assess the characteristics of the dysfunction of semicircular canal in benign paroxysmal positional vertigo and the relationship with the ectopic otoconia.</p><p><b>METHODS</b>There were 214 patients with benign paroxysmal positional vertigo (BPPV), including 107 cases of posterior semicircular canal canalithiasis (PSC-Can) 80 cases of horizontal semicircular canal canalithiasis (HSC-Can), 27 cases of horizontal semicircular canal cupulolithiasis (HSC-Cup). One hundred and ninety (88.8%) patients were accompany with relevant diseases while 24 (11.2%) cases were not. They accepted low, middle and high frequency vestibular function tests, including caloric test (CT), head shaking test (HST) and video head impulse test (vHIT) respectively. The parameters of the unilateral weakness (UW), head shaking nystagmus (HSN) and video head impulse test gain (vHIT-G) were observed. Patients classified into three groups (PSC-Can, HSC-Can, HSC-Cup) according to the involvement semicircular canal. The results of the three tests were analyzed with SPSS16.0 software.</p><p><b>RESULTS</b>The positive cases of the three tests were vHIT: 15 (7.0%), HST: 52 (24.3%), CT: 152 (71.0%), a statistically significant difference (P < 0.05) was found between the three tests. When compared the Caloric Test, HST and vHIT between the BPPV patients with and without relevant diseases, there were no significant differences (P > 0.05). The variance without statistical significance (P > 0.05) was showed between three tests' results in each groups, it was also showed that the variance between the three groups in each tests reached no statistical significance (P > 0.05). The test of affected side UW between PSC-Can, HSC-Can and HSC-Cup showed the variance without statistical significance (F = 0.970, P = 0.383).</p><p><b>CONCLUSIONS</b>The lesion of semicircular canals has the same etiological factors with the utricle pathological change in benign paroxysmal positional vertigo, and the dysfunction mostly happens in low frequency range of semicircular canal frequency band. The ectopic otoconia is not the main etiological factors for that. HST and vHIT of middle and high frequency can not be ultimately used for the screening test evaluating due to the semicircular canal function in BPPV.</p>

Morphologic Change of the Vestibular Organ in the Na+-K+-2Cl- Cotransporter Deficiency Mouse

BACKGROUND AND OBJECTIVES: The Na+-K+-2Cl- cotransporter-1 (NKCC1) is a member of the cation-coupled chloride transporter that participates in salt transport and cell volume regulation in diverse tissues. NKCC1 deficient mice exhibit deafness, and have structural alterations in the cochlea. In addition to hearing loss, NKCC1-deficient mice show a shaker-waltzer behavior, which suggests a vestibular system defect. This study investigated the morphology of the vestibular system of NKCC1-deficient mice. In addition, this study evaluated whether NKCC1 mRNA and its protein are expressed in human vestibular end organs. MATERIALS AND METHOD: NKCC1-deficient and wild type mice aged 4~5 weeks were sacrificed. Their heads were cut in the midsagittal plane, fixed and decalcified. For light microscopy, 5 m sections were cut, and stained with hematoxylin and eosin. Human vestibular end organs were harvested during acoustic tumor surgery via translabyrinthine approach. Some of these end organs were used for the total mRNA extraction and the remainder was used for immunostaining. RT-PCR was performed for NKCC1. RESULTS: The scala media of the cochlear of the NKCC1-deficient mice were collapsed but the bony labyrinth of the cochlea appeared unaffected. However, the semicircular canals (SCCs) were much smaller than those in the wild type. Furthermore, the SCCs were completely missing in some NKCC1-deficient mice. NKCC1 mRNA was expressed in both human macula and crista ampullaris and its protein was expressed mainly in the transitional and dark cell area of the human crista ampullaris. CONCLUSION: NKCC1 may be essential for maintaining the vestibular morphology and its function in mice and NKCC1 is well expressed in human vestibular end organs.

Morphologic Change of the Vestibular Organ in the Na+-K+-2Cl- Cotransporter Deficiency Mouse

BACKGROUND AND OBJECTIVES: The Na+-K+-2Cl- cotransporter-1 (NKCC1) is a member of the cation-coupled chloride transporter that participates in salt transport and cell volume regulation in diverse tissues. NKCC1 deficient mice exhibit deafness, and have structural alterations in the cochlea. In addition to hearing loss, NKCC1-deficient mice show a shaker-waltzer behavior, which suggests a vestibular system defect. This study investigated the morphology of the vestibular system of NKCC1-deficient mice. In addition, this study evaluated whether NKCC1 mRNA and its protein are expressed in human vestibular end organs. MATERIALS AND METHOD: NKCC1-deficient and wild type mice aged 4~5 weeks were sacrificed. Their heads were cut in the midsagittal plane, fixed and decalcified. For light microscopy, 5 m sections were cut, and stained with hematoxylin and eosin. Human vestibular end organs were harvested during acoustic tumor surgery via translabyrinthine approach. Some of these end organs were used for the total mRNA extraction and the remainder was used for immunostaining. RT-PCR was performed for NKCC1. RESULTS: The scala media of the cochlear of the NKCC1-deficient mice were collapsed but the bony labyrinth of the cochlea appeared unaffected. However, the semicircular canals (SCCs) were much smaller than those in the wild type. Furthermore, the SCCs were completely missing in some NKCC1-deficient mice. NKCC1 mRNA was expressed in both human macula and crista ampullaris and its protein was expressed mainly in the transitional and dark cell area of the human crista ampullaris. CONCLUSION: NKCC1 may be essential for maintaining the vestibular morphology and its function in mice and NKCC1 is well expressed in human vestibular end organs.

Three Dimensional Analysis of Caloric, Spontaneous, Positional, Positioning and Post Head Shaking Nystagmus in Bilateral Vestibulotoxic Cats / 대한이비인후과학회지

BACKGROUND AND OBJECTIVES: Vestibular toxicity can be usually monitored by a caloric test, but measuring eye movements including nystagmus is also helpful in the evaluation of vestibulotoxic process. To understand the vestibular signs during bilateral vestibulotoxic process, we observed three dimensional serial changes of eye movements and the results of caloric test in cats using a magnetic search coil system. MATERIALS AND METHODS: Three-dimensional (3-D) eye movement responses to ice water caloric stimulus and spontaneous, positional, positioning, and post head shaking nystagmus in five cats were serially evaluated at 3, 7, 10, 14, 21, 28 days, 2, 3, 4, 5, 6 months after inducing bilateral vesitibulotoxicity by subcutaneous injection of streptomycin. Histologic changes of vestibular system were also observed. RESULTS: Bilateral vestibulotixicity which showed no response to ice water caloric stimulus was induced within 2 weeks with streptomycin injection. Positional, positioning and post head shaking nystagmus were present during vestibulotoxic process. Caloric nystagmus did not reappear within 6 months and histologic finding in crista ampullaris of lateral semicircular canal showed marked degeneration of the hair cells and nerves. CONCLUSION: Observation of positional, positioning and post head shaking nystagmus as well as ice water caloric test will be helpful to monitor vestibulotoxic process.

Human Embryos of Carnegie Stage 17 / 대한해부학회지

Morphological characteristics of human embryos of Carnegie stage 17 were described. The crown rump length of these embryos were 11.7-13.6mm. These embryos were characterized externally by ventrally directed nasal pits, 6 distinct auricular hillocks, finger rays in the hand plate, digital plate at the lower limb bud, herniation of a part of midgut through umbilical cord, and internally by the partial fusion of ventral and dorsal pancreas, prominent nasal sac which is not open to primitive oral cavity, formation of segmental and some subsegmental bronchial buds, branching of metanephrotic pelvis, crescent-shaped lens cavity, and imminent semicircular ducts.