The Expression of Cav1.3 Calcium Channels in Adult Rat Cochlea / 听力学及言语疾病杂志

Objective To investigate the expression of Cav 1.3 calcium channel in adult rat cochlea and study its role in auditory physiology and pathology.Methods The sprague-dawley rats were used as experimental subjects.The distribution of Cav1.3 calcium channel in the cochlea was detected by immunofluorescence technique.The expression of Cav1.3 was measured with Western blot (WB) and RT-PCR.Results Immunofluorescence photographs revealed that Cav 1.3 calcium channel localized in the lateral wall membrane,hair cells,stria vascularis,spiral ganglion cell,spiral ligment,spiral prominence,and limbus laminae spiralis.The results of WB and RT-PCR inform Cav1.3 calcium channel gene (CACNA1D) were measured in the cochlea and kidney.The expression of Cav1.3was mainly in the basilar membrane.Moderate expression was observed in the spiral ganglion and stria vascularis.Conclusion The preliminary study revealed the distribution of Cav 1.3 calcium channel gene(CACNA1D)in adult rat cochle possesses tissue specificity,providing a theoretical basis for further research in auditory physiology and pathology.

The Expression of Plasma Membrane Ca2 + -ATPase Isoforms 1~4 and the Splice Variants at Sites A and C in the Neonatal Rat Vestibular Organ / 听力学及言语疾病杂志

Objective To study the expression of plasma membrane Ca2 + -ATPase isoforms 1 -4 and the splice variants at sites A and C in the neonatal rat vestibular organ.Methods Ten rats at postnatal 2 days (P2 ) were decapitated and their vestibular organs (macula utriculi and macula sacculi)were isolated.The total proteins of the vestibular organs were extracted.The expression of PMCA1-4 splice variants at sites A and C was detected by RT-PCR.Results The splice variants of PMCA1-4 at sites A and C in macula utriculi and macula sacculi of neo-natal rat vestibular organs were PMCA1x/b,PMCA2w/(a,b),PMCA3z/(a,b,c)and PMCA4 (x,z)/b.Conclusion The splice variants at sites A and C among PMCA1,PMCA2,PMCA3 and PMCA4 were different in the vestibu-lar organs of neonatal rats,which could be explained that macula utriculi and macula sacculi had different require-ments of Ca2 + turning for these PMCA isoforms.

The Expression of Plasma Membrane Ca2 + -ATPase Isoforms 1 ~3in the Basi lar Membrane of Neonatal Rat Cochlea / 听力学及言语疾病杂志

Objective To study the expression of plasma membrane Ca2+-ATPase isoforms 1~3 (PMCA 1~3 )in the basilar membrane (BM)of the neonatal rat cochlea by Western blot.The PMCA2 content in single BM of the neonatal rat was also examined.Methods Four rats at postnatal 2 days (P2)and 8 days (P8)were respective-ly decapitated and their BMs were isolated.The total proteins of BMs were extracted.The 20μg total proteins were respectively loaded to the gel.The expression of PMCA1-3 was detected by Western blot.Likewise,3μg total proteins from P2 and P8 rat BM were loaded.The expression of PMCA2 was detected by Western blot.Four rats at P8 were decapitated and their BM was isolated.The 5μg,10μg and 20μg total proteins of P8 rat BM were added to the gel and 100 ng,400 ng and 800 ng bovine serum albumin (BSA)were also loaded as reference.After electro-phoresis,the gel was separated into two parts.One part was used for SYPRO staining and the other part was used for PMCA2 detection by Western blot.Results In the 20μg BM total proteins of P2 and P8 rats,the expression of PMCA1 was weak (0.126±0.024,0.131±0.012,respectively),PMCA2 was strong (4.16±0.528,4.25±0.319, respectively),and PMCA3 was barely expressed (0 ).There was a statistical difference among PMCA1 ,PMCA2 and PMCA3(P<0.05).In the 3μg BM total proteins of P2 and P8 rats,the expression of PMCA2 in P8 (4.571± 0.336)was higher than P2 (3.622±0.285).There was a statistical difference(P<0.05).The PMCA2 content in the BM of a P8 rat was about 2 .5 ng.Conclusion There was a different-level expression of PMCA1~3 in the neonatal rat BM with highest expression of PMCA2 ,which could be explained that cochlear hair cells had different requirements of Ca2+ turning for these PMCA isoforms.

Age-related expression of plasma membrane Ca(2+) -ATPase isoform 2 in the cochleas of C57BL/6J mice / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To investigate the location and distribution of plasma membrane Ca²⁺ -ATPase isoform 2(PMCA2) in the cochleas of C57BL/6J mice at various ages (4w, 14w, 22w, 45w), and to reveal the relationship of PMCA2 and age-related hearing loss (AHL).</p><p><b>METHODS</b>The distribution of PMCA2 in the cochleas of C57BL/6J mice was detected by immunohistochemistry at various ages (4w, 14w, 22w, 45w). Real-time polymerase chain reaction (Rt-PCR) was used to detect the level of PMCA2 mRNA in the cochleas of C57BL/6J mice at the ages of 4, 14, 22 and 45 weeks old respectively. Using SPSS17.0 software for statistical analysis.</p><p><b>RESULTS</b>PMCA2 was mainly located in the hear cells, stria vascularis, and spiral ganglion cells. Faint labeling of PMCA2 was also observed in spiral ligament. Hair cells missed and the number of spiral ganglion cells reduced with age. Expression of PMCA2 in the cochleas of C57BL/6J mice also showed age-related decreasing. The results of Rt-PCR demonstrated the expression of mRNA of gene (Atp2b2) at 14 weeks age was significantly less than 4 week-old mice cochlears (P<0.05). The expression of mRNA of gene (Atp2b2) at 22 weeks age was significantly less than 14 week-old mice cochlears (P<0.05). The expression of mRNA of gene (Atp2b2) at 45 weeks age was significantly less than 14 week-old mice cochlears (P<0.01).</p><p><b>CONCLUSIONS</b>PMCA2 is mainly located in the hear cells, stria vascularis, and spiral ganglion cells. Faint labeling of PMCA2 is also observed in spiral ligament. The expression of PMCA2 demonstrates an age-related decrease with age. The mRNA expression level of PMCA2 gene(Atp2b2) in the cochleas of C57BL/6J mice displayed an age-related decrease. PMCA2 transporters may play a critical role in maintaining the normal morphology of the inner ear and it may be related to AHL.</p>

Causes of the recurrence of benign, paroxysmal positional vertigo / 中华物理医学与康复杂志

Objective To investigate why benign, paroxysmal positional vertigo (BPPV) recurs.Methods Three hundred persons diagnosed with BPPV who had been treated at Tongji Hospital of Huazhong University of Science and Technology between April 2012 and April 2014 were given a telephone follow-up at least one year after their manual repositioning treatment.The respondents were divided into a healthy group and a recurrence group according to whether they said their vertigo had recurred.The age and gender distributions of the two groups were compared, along with their underlying diseases and living-related factors.Causes of the recurrence were then hypothesized.Results Single factor analysis and binary logistic regression analysis showed that overwork, an age over 45, travelling frequently, long use of computers, sleep disorders, oral intake of calcium tablets, posterior circulation ischemia and hyperlipidemia were all closely related to the BPPV recurrence.Age over 45 showed the strongest correlation.Conclusion Aging is the greatest risk factor for the recurrence of BPPV.Posterior circulation ischemia, hyperlipidemia, overwork, sleep disorders, long use of computers and being on business frequently are also predictors of relapse.

Correlation of PDCD5 and apoptosis in hair cells and spiral ganglion neurons of different age of C57BL/6J mice / 华中科技大学学报（医学）（英德文版）

This study examined the expression pattern of programmed cell death 5 (PDCD5) in cochlear hair cells and spiral ganglion neurons (SGNs) and its association with age-related hearing loss in mice. Sixty C57BL/6J (C57) mice at different ages were divided into four groups (3, 6, 9 or 12 months). PDCD5 expression was detected by using immunohistochemistry, real-time PCR and Western blot. Morphological change of the cochleae was also evaluated by using immunoassay. The results showed that the expression of PDCD5 had a gradual increase with ageing in both protein and RNA levels in C57 mice, as well as gradually increased apoptosis of cochlear hair cells and SGNs. In addition, we also found that caspase-3 activity was enhanced and its expression was enhanced with ageing. It is implied that overexpression of PDCD5 causes the increase in caspase-3 activity and the subsequent increase of apoptosis in cochlear hair cells and SGNs, and thereby plays a role in the pathogenesis of presbycusis. Thus, PDCD5 may be a new target site for the treatment and prevention of age-related hearing loss.

The study on growth and proliferation of neural stem cells from rats in vitro / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To investigate culturing neural stem cells (NSCs) from rat embryos in vitro and to observe their growth and differentiation.@*METHOD@#NSCs were isolated from hippocampus of SD rat embryos (P16-P18) and cultured in DMEM/F12 medium containing EGF, bFGF, B27. To observe process of cell proliferation by microscope and identify cell types by immunocytochemical analyses after differentiation.@*RESULT@#NSCs grew well in serum-free conditional medium and their cell bodies present transparent with good refraction at about eighth day. After differentiation, the cells demonstrated NSE and GFAP immunoreactive.@*CONCLUSION@#NSCs were cultured well in serum-free conditional medium and they could be induced to differentiate into neurons and astrocytes in serum conditional medium.

Effect of the intense noise on hearing function and cochlea morphology in rat / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To observe the changes of auditory electrophysiology and inner ear pathology in rat cochlea after the noise exposure, and to offer the experimental data for exploring the mechanism of noise-damaged cochlea.@*METHOD@#The rats in the study group were exposed to a intense narrow band noise centered at 4 kHz at the leave of 120 dB (SPL) for 4 h. The exposed cochleae were collected at various intervals (1 or 21 days) after the noise exposure. Auditory function was monitored by measuring thresholds of auditory brain stem responses (ABR). The morphological changes in rat cochlear hair cell (HC) were examined by HC nuclei stained with Propidium iodide (PI), a fluorescent dye specifically labelling the nuclear DNA and scanning electron microscopy (SEM). The number of spiral ganglion cells was calculated using pathologic technique.@*RESULT@#The thresholds of ABR in the study group were significantly greater than that in the normal control group (P 0.05). SEM revealed the injured stereocilia of OHC (disarrangement, collapse) and OHC loss in the study group, which was more severe in OHC3 than the other two rows of OHC.@*CONCLUSION@#The intense noise used in our study could injure the rat cochlea and bring permanent threshold shift (PTS). Under this condition, the death modes of HC in the cochlea include apoptosis and necrosis in the fore part, whereas necrotic is the major mode in the evening of exposure. The injured stereocilia of OHC and OHC loss could remain the most consistent correlate of PTS.

Study on the TIM-4 gene polymorphism and its association with the susceptibility to allergic asthma in Chinese Ban population of Hubei province / 中华微生物学和免疫学杂志

Objective To investigate whether two polymorphism sites of the the exon 2 Lys65Lys(G/A) and exon 9 Va1365Met(G/A) in T ceils immunoglobulin domain and mucin domain protein-4(TIM-4) are associated with asthma in Chinese Han population of Hubei province. Methods The polymorphisms were de-tected with polymerase ehain reaction-restriction fragment length polymorphism(PCR-RFLP) in 185 cases of al-lergic asthma and 162 healthy controls. The genotype and allele frequencies were calculated and analyzed. Re-sults(1)The genotype frequencies of G/G, G/A and A/A in Lys65Lys(G/A)polymorphism were 0.840, 0.160 and 0 respectively in the healthy population, and were 0.859, 0.141 and 0 respectively in the allergic asthma population. No significant difference in genotype and allele frequencies was found between asthma pa-tients and the control subjects (P=0.603, P=0.618). (2) The polymorphism of the Val365 Met(G/A) was not detected in our study. Conclusion There is polymorphism site of the exon 2 Lys65Lys(G/A)in TIM-4, but this polymorphism site is not associated with asthma in Han nationality in Hubei Chinese population. There is no SNP of the exon 9 Val365Met(G/A) in TIM-4 in Chinese Han population of Hubei province.