Yinyan Hu

Chinese PLA General Hospital (301 Hospital), Peping, Beijing, China

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Publications (8)5.58 Total impact

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    ABSTRACT: To observe whether bFGF could cross the blood-labyrinth barrier (BLB) after intra-abdominal injection and to establish an experimental basis for its clinical applications. Thirty guinea pigs were divided into three groups. Animals in group 1 were administered o I-bFGF, while animals in group 2 and 3 were administered 125 and saline, respectively, via intra-abdominal injection. The both cochlea, blood, liver, brain, thyroid gland and kidney were collected and weighted. A radioimmunoassay analyzer was employed to measure counts per minute (CPM) of each sample, and autoradiography was performed on both cochlea. The CPM value of organ samples in the 125I group was higher than that in other groups, and radioactive grain was observed in cochlear samples of this group. In the 125I-bFGF group, blood demonstrated the highest CPM value, while cochlea and brain demonstrated the lowest CPM value, with no radioactive grain observed in cochlear samples. bFGF has some difficulties in getting across BLB, so the way of bFGF application in clinics need further study.
    Lin chuang er bi yan hou ke za zhi = Journal of clinical otorhinolaryngology 08/2010; 24(16):750-2.
  • Qi Li, Lin Lin, Yinyan Hu, Deliang Huang
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    ABSTRACT: To detect the expression of Aquaporin-1,3 in the cochlea and endolymphatic sac of guinea pig. Two-step immunohistochemical method and immunofluorescence was used to examine the expression of Aquaporin-1,3. Aquaporin-1 was expressed on the basilar part of spiral ligament, basal membrane of Corti's organ and epithelialis of scala tympani, and basilar part under the cellular epithelialis in endolymphatic sac. Aquaporin-3 was expressed on stria vascularis, spiral ligament, Corti's organ, spiral ganglion and, the basilar part and the cellular epithelialis in endolymphatic sac. Aquaporin-1,3 were widely expressed in the cochlea and endolymphatic sac of guinea pig.
    Lin chuang er bi yan hou ke za zhi = Journal of clinical otorhinolaryngology 01/2006; 19(23):1085-7.
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    ABSTRACT: Cochlear hair cells are a terminally differentiated cell population that is crucial for hearing. Although recent work suggests that there are hair cell progenitors in postnatal mammalian cochleae, isolation and culture of pure hair cell progenitors from a well-defined cochlear area have not been reported. Here we present an experimental method that allows isolation and culture of hair cell progenitors from postnatal rat cochleae. These progenitor cells are isolated from the lesser epithelial ridge (LER, or outer spiral sulcus cell) area of pre-plated neonatal rat cochlear segments. They express the same markers as LER cells in vivo, including ZO1, Islet1, Hes1, and Hes5. When these cells are induced to express Hath1, they show the potential to differentiate into hair cell-like cells. Interestingly, these cells can be lifted from monolayer cultures and maintained in aggregate cultures in which spheres can be formed. Hair cell progenitors in the spheres display their proliferating capability and express only epithelial markers. Furthermore, when these spheres are mixed with dissociated mesenchymal cells prepared from postnatal rat utricular whole mounts, and replated onto a collagen substratum, the epithelial progenitor cells are able to differentiate into cells expressing markers of hair cells and supporting cells in epithelial islands, which mirrors the inner ear sensory epithelium in vivo. Successful isolation and culture of hair cell progenitors from the mammalian cochlea will facilitate studies on gene expression profiling and mechanism of differentiation/regeneration of hair cells, which are crucial for repairing hearing loss.
    Journal of Neurobiology 01/2006; 65(3):282-93. · 3.05 Impact Factor
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    ABSTRACT: To investigate the method for isolating greater epithelial ridge by use of thermolysin digestion combined with dissection under microscope. The basement membrane prepared from postnatal day 0 to postnatal day 3 rat was placed into D-Hanks solution with thermolysin and DNase, then incubated at 37 degrees C for 25 minute and dissected under microscope. After GER had been isolated, morphological comparison, immunohistochemistry, RT-PCR and GER culture were performed for the purpose of identification of GER. Immunohistochemistry indicated that the isolated GER was purely epithelial tissue. RT-PCR analysis certified that the isolated GER expressed Hes1 which was selectively expressed in GER at early postnatal in cochlea. The isolated GER could be successfully cultured in the presence of 5% serum and could survive at least 12 day in the serum-free medium. GER isolation can be successfully performed by use of thermolysin combined with microdissection. This method may provide a good technique to establish the GER cell lines or perform series of gene transfection experiments.
    Lin chuang er bi yan hou ke za zhi = Journal of clinical otorhinolaryngology 08/2005; 19(13):611-3.
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    ABSTRACT: To investigate the molecular basis of the voltage-gated Ca2+ channel (VGCC) which controls the afferent synaptic neurotransmitters release in inner hair cell and their cellular expression. In this study we used different and complementary approaches to determine which types of 4 subunits of VGCC were expressed in the organ of Corti. RT-PCR analysis showed that alpha1C and alpha1D were present in the organ of Corti total RNA extract, and the corresponding antisense riboprobes could detect the expression of alpha1C and alpha1D in IHCs and OHCs. The voltage-gated Ca2+ channel expressed in sensory cell in organ of Corti are alpha1C and alpha1D subtypes.
    Lin chuang er bi yan hou ke za zhi = Journal of clinical otorhinolaryngology 11/2004; 18(10):613-5.
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    ABSTRACT: Preventing noise-induced hearing loss (NIHL) by antioxidants is based on the hypothesis that generation of reactive oxygen species is one of the causes of NIHL. alpha-Tocopherol is a naturally occurring antioxidant with no noticeable side effects. In this study, we attempted to protect guinea pigs from developing NIHL by administering alpha-tocopherol. Pigmented male guinea pigs were exposed to a noise (4 kHz octave band, 100 dB SPL), 8 h/day for 3 days consecutively. alpha-Tocopherol (10 mg/kg or 50 mg/kg daily) was given by intraperitoneal injection from 3 days before through 3 days after the noise exposure. Auditory evoked brainstem response (ABR) thresholds at 2, 4 and 8 kHz were recorded prior to the experiment, immediately post-noise, 2 and 8 days post-noise. On day 8 post-noise, after the ABR recording, guinea pigs were decapitated and the cochleae were removed for cochlear surface preparations and scanning electron microscope (SEM) study. ABR threshold shifts of groups receiving alpha-tocopherol were significantly smaller than those of groups not receiving alpha-tocopherol at all frequencies and all time points tested except that of group 3 at 8 kHz 8 days post-noise. No hair cell loss was seen on the surface preparations, but stereocilia loss was found by SEM study. The noise-induced stereocilia loss was significantly decreased by alpha-tocopherol. These results indicate that alpha-tocopherol can attenuate the noise-induced cochlear damage. Further investigations on the preventive effect of alpha-tocopherol on NIHL in noise-exposed workers are necessary.
    Hearing Research 06/2003; 179(1-2):1-8. · 2.54 Impact Factor
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    ABSTRACT: To observe the expression of cationic stearylamine (SA) liposome mediated basic fibroblast growth factor/green fluorescence protein (bFGF/GFP) gene in the cochlea of guinea pig, and evaluate the protection and rescue action of bFGF against the damage caused by gentamicin. Thirty-six guinea pigs were divided into 3 experimental groups. Prevention group with inoculation of SA-bFGF/GFP complexes through the round window of right ear and injection of gentamicin 150 mg.kg(-1).d(-1) one day after for 8 days and rescue group with injection of gentamicin for 8 days and infusion of SA-bFGF/GFP complexes in the same way on the ninth day, and control group with only injection of gentamicin for 8 days. Auditory brainstem responses (ABR) was measured prior to and after the administration and before the animals were killed respectively. The animals were killed after the experiment and ABR test, and specimens and slices of chochleae were made to examine the absence of outer and inner hair cells. The expression of GFP in cochlea shown by green fluorescence was observed with fluorescent microscopy. Fluorescent microscopy showed green fluorescence in the cochleae of guinea pigs in prevention and rescue groups. There was no significant difference in ABR threshold between the left and right ears of animals in each group before and after experiment, among both ears of animals in the 3 groups before experiment, and between prevention and rescue group groups before killing (all P > 0.05). However, the ABR thresholds in prevention group and rescue group were significantly lower than that in control group before the animals were killed (P < 0.01 and P < 0.05). The average number of lost outer hair cells was 5 106 +/- 299 cells and 5 605 +/- 109 cells in prevention group and rescue group respectively, without a significant difference between them (P > 0.05). The average amount of missing inner hair cells was 301 +/- 64 cells and 487 +/- 92 cells in prevention group and rescue group respectively, without a significant difference between them (P > 0.05), and significantly lower than that in control group (1 062 +/- 67, P < 0.01 and P < 0.05). The average amount of missing outer hair cells in control group was 6 248 +/- 119 cells, significantly higher than that in prevention group (5 106 +/- 299, P < 0.01) and that in rescue group (5 605 +/- 109, P < 0.05). SA-liposome mediated bFGF/GFP gene, which was perfused in one ear, can be expressed highly in both cochleae of the guinea pig, and may protect and rescue cochlea against gentamicin ototoxicity.
    Zhonghua yi xue za zhi 09/2002; 82(17):1192-4.
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    ABSTRACT: To observe the expression of brain derived neurotrophic factor (BDNF) in the cochlea of guinea pig, and assess the activity of BDNF in spiral ganglion cell following the damage of noise. Twenty-seven guinea pigs were exposed to a 4 kHz narrow band noise at 135 dB SPL for 4 hours. At seven days after the noise exposure, twelve guinea pigs were inoculated with ad-BDNF, and twelve guinea pigs were inoculated with ad-LacZ, while three guinea pigs were inoculated with artificial perilymphatic fluid. The animals were sacrificed After 1 week, 4 weeks and 8 weeks, respectively. The cochlea was stained with immunohistochemisty (ABC method) to examine the expression of BDNF. The number of spiral ganglion cell was counted to assess the activity of BDNF. BDNF gene was expressed in whole cochlea, and the expression was obviously elevated 1 week after the adenovirus administration and reduced thereafter. The number of degenerated cells in ad-BDNF group in the eight weeks was lower than that in another two groups and the different between them was statistical significant (P < 0.01). Adenoviral-mediated brain derived neurotrophic factor can be expressed in a high level in the cochlea of guinea pig, and may prevent the cell of spiral ganglion from the damage of noise. This study lays the groundwork for alleviation of hearing loss using gene therapy.
    Zhonghua er bi yan hou ke za zhi 04/2002; 37(2):109-11.