Pei Chen

Capital Medical University, Peping, Beijing, China

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Publications (18)4.69 Total impact

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    ABSTRACT: We aimed to compare the characteristics between lentivirus and adenovirus vector mediated gene transfer into cultured spiral ganglion cells (SGCs). SGCs from newborn rats were cultured and exposed to lentivirus-GFP and adenovirus-GFP vectors. GFP expression and the cell morphology were evaluated under epi-fluorescence microscope at 3 days and 7 days after exposure. Survival number of SGCs was counted, and the average percentage of SGCs with GFP expression was calculated, and axon length was measured by ImageJ software. Cultured SGCs were transfected by either adenovirus or lentivirus vector successfully. The adenovirus vector presented an instant and efficient transfection. However, the expression of GFP went down after 7 days. In lentivirus-GFP group, GFP expression was detected at 7 days after exposure, and the number of cells with GFP expression increased gradually in the following days. Statistical analysis revealed that there were no differences in survival number of SGCs and average axon length among lentivirus-GFP group, adenovirus-GFP group and control group. Cultured SGCs can be transfected by either lentivirus vector or adenovirus vector safely and efficiently. SGCs are more susceptible to adenovirus vector, but GFP persists for a longer period after the lentivirus-mediated gene transfer.
    Lin chuang er bi yan hou ke za zhi = Journal of clinical otorhinolaryngology 02/2011; 25(4):172-5.
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    ABSTRACT: Penetrating cranionasal injuries are relatively rare, usually occur in young children, and can be caused by a variety of unusual objects. The mortality and disability rates are high without appropriate treatment. We report a penetrating cranionasal injury caused by a bicycle spoke. Some fundamental principles in the diagnosis and treatment of penetration injuries are emphasized from this case and the surrounding literature reviewed.
    Pediatric emergency care 11/2010; 26(11):837-9. · 0.92 Impact Factor
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    ABSTRACT: Both reinnervated and dormant neurons can be observed in facial nucleus following facial-facial anastomosis. Although they may play different roles in the remodeling mechanism of facial nucleus during facial nerve injury and regeneration, comprehensive gene profiling analysis of these two neuron types has never been performed due to the difficulty in isolating specific neuron populations and extracting sufficient amount of RNAs from the heterogeneous facial nucleus. In this study, we developed a method to isolate the Fluoro-Ruby (FR) retrogradely labeled reinnervated facial motor neurons and the Nissl stained dormant neurons in two steps with laser capture microdissection (LCM) technology. The quality and yield of RNAs extracted from these isolated neurons were confirmed with Agilent 2100 Bioanalyzer. This modified LCM based neuron isolation method will facilitate the studies of the roles and interactions of different neurons that co-exist in the same facial nucleus after peripheral injury.
    Neuroscience Letters 11/2009; 468(3):316-9. · 2.03 Impact Factor
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    ABSTRACT: To make clear the molecular pathways involved in hydrogen peroxide-induced spiral ganglion cells death. The spiral ganglion cells of the newly born rats were primary cultured. Then the SGCs were exposed to hydrogen peroxide for different concentrations (0, 100, 200, 500 micromol/L) and for different hours (2, 4, 6 h). Cell nucleic were stained simultaneously with the DNA binding dyes Hoechst 33258 and propidium iodide. At lower concentrations of hydrogen peroxide, apoptosis was the main reason for cell death. At higher concentrations of hydrogen peroxide, the cells died mainly by necrosis. The effects of hydrogen peroxide are dose and time dependency. Reactive oxygen species may play a role as an early molecule signal in the mechanism of SGCs death.
    Lin chuang er bi yan hou ke za zhi = Journal of clinical otorhinolaryngology 04/2009; 23(5):222-4.
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    ABSTRACT: The remodeling process of synapses and neurotransmitter receptors of facial nucleus were observed. Models were set up by facial-facial anastomosis in rat. At post-surgery day (PSD) 0, 7, 21 and 60, synaptophysin (p38), NMDA receptor subunit 2A and AMPA receptor subunit 2 (GluR2) were observed by immunohistochemical method and semi-quantitative RT-PCR, respectively. Meanwhile, the synaptic structure of the facial motorneurons was observed under a transmission electron microscope (TEM). The intensity of p38 immunoreactivity was decreased, reaching the lowest value at PSD day 7, and then increased slightly at PSD 21. Ultrastructurally, the number of synapses in nucleus of the operational side decreased, which was consistent with the change in P38 immunoreactivity. NMDAR2A mRNA was down-regulated significantly in facial nucleus after the operation (P<0.05), whereas AMPAR2 mRNA levels remained unchanged (P>0.05). The synapses innervation and the expression of NMDAR2A and AMPAR2 mRNA in facial nucleus might be modified to suit for the new motor tasks following facial-facial anastomosis, and influenced facial nerve regeneration and recovery.
    Journal of Huazhong University of Science and Technology 12/2008; 28(6):714-8. · 0.58 Impact Factor
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    ABSTRACT: To observe the recovery process of facial behavior and function in rat, and then to supply reliable functional parameters for the researches in such fields. Rat models of facial nerve paralysis were set up by sectioning and anastomosis of facial nerve. The behavioral change included whisker movement and blink reflex were observed weekly. Electroneurography (ENoG) and blink reflex (BR) were examined dynamically and all data were analyzed by statistic soft ware. Postoperatively, the whisker movement ceased, blink reflex was lost or sluggish but the fibrillation of vibrissae appeared. Whisker movement and evoke blink reflex were seen 1-2 months following operation gradually, which subsequently increased in intensity and frequency. Mass contraction of the periauricular muscles were observed at the same time as eye closure 2 month following operation. The latency of compound muscle action potential (CMAP) at experimental side began to prolong at 21 day, reached climax at 1 month and was stabilized at 3-4 month postoperatively, but it could not get full recovery. The latencies of 28-63 day were longer than other time points (P<0.05). The amplitude and intensity didn't change characteristically. The R1 can be observed repetitively, which disappeared at 7-14 day and gradually recovered 1 month following operation. At experimental side, the R1-type wave (R1oris) in orbicularis oris could be observed at the same time as R1 recorded 2 month following operation, which indicated the facial synkinesis, one hyperkinetic post-paralytic sequela happened. Then the latency of both R1 and R1oris decreased concomitantly. There were correlations between them, but only the significant difference of R1oris latency presented between 2 month and other time points (P<0.05). It is concluded that the methods of ENoG and BR could examine the recovery process of facial movement, which would help studying the pathophysiological mechanism of facial nerve injury and regeneration after being revised.
    Lin chuang er bi yan hou ke za zhi = Journal of clinical otorhinolaryngology 04/2008; 22(7):318-21.
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    ABSTRACT: To purify P0 protein from guinea pig's inner ear by preparative SDS-PAGE and study the possible role it may play in the etiology of autoimmune inner ear disease. A mixture of membraneous proteins of inner ear was separated by preparative SDS-PAGE. The corresponding band at 30kd was cut and electrically eluted. The protein collected was identified by analytical SDS-PAGE and Western blot assay. A group of 20 guinea pigs were immunized with P0 protein emulsified in complete Freund's adjuvant, another 10 guinea pigs were immunized with complete Freund 's adjuvant only as control. The guinea pigs' hearing thresholds, serum IgG level and morphological changes in the inner ear were investigated. The distribution of P0 protein in the cochlear was detected by immunohistochemical technique. The purity of the protein was demonstrated by a single band at the 30 kD site in SDS-PAGE, which was identified as P0 protein by western blot analysis assay. About 17.5% P0-immunized guinea pigs showed increased hearing thresholds, elevated IgG level (F =6.48, P <0. 01), as well as a decreased number of spiral ganglion cells and inflammatory cell infiltration in the cochlear nerve region. The P0 protein is distributed in the cochlear nerve and spiral ganglion only. P0 protein from guinea pig's inner ear can be successfully purified by preparative SDS-PAGE and an animal model of experimental autoimmune inner ear disease induced by P0 protein is successfully established.
    Lin chuang er bi yan hou ke za zhi = Journal of clinical otorhinolaryngology 08/2007; 21(16):748-51.
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    ABSTRACT: To study the suppression of proliferation of Hep-2 cells by stable expression of anti-sense perlecan cDNA. In this study, the plasmids of recombination eukaryotic expression vector perlecan anti-sense cDNA (pAP) were transfected into Hep-2 cells by using cationic liposome (lipofectamine 2000) and divided into three groups: non-transfected group, WT group; transfection with no load carrier, neo group; and transfection with the pAP plasmid, pAP group. Semi quantify RT-PCR, western blot assay and MTT assay were used to detected the expression of perlecan mRNA and protein in the three groups; the level of cell proliferation; and the responsivity of basic fibroblast growth factor (bFGF). It was showed that the expression of perlecan mRNA and protein were significantly reduced in the pAP group compared with WT group and ph beta Apr-neol transfected group ( P < 0.01). In the presence of 1 microg/L of bFGF in low serum (0.1% FCS), the pAP transfected cells showed a reduced proliferation rate (MTT assay) while the wild type cells and ph beta Apr-neol transfected cells grew rapidly. The growth of Hep-2 cells could be inhibited significantly by perlecan anti-sense cDNA plasmids transfection.
    Lin chuang er bi yan hou ke za zhi = Journal of clinical otorhinolaryngology 11/2006; 20(21):984-7.
  • Zhonghua er bi yan hou tou jing wai ke za zhi = Chinese journal of otorhinolaryngology head and neck surgery 10/2006; 41(9):643.
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    ABSTRACT: To recognise the predisposing factors, clinical manifestations, diagnosis and treatment of neurofibromatosis type 2 (NF2). The clinical data of one NF2 case was reported and the literatures were also reviewed. The patient was diagnosed at a much later stage than onset. Progressive hearing loss and tinnitus were the initial symptoms. MRI scan indicated space-occupying lesions in the bilateral cerebellopontine angles, bilateral cavernous sinuses and bilateral cervical parts of the patient. The patient was diagnosed as NF2 according to the National Institutes of Health (NIH) criteria, and received operation on the left acoustic tumor. The tumor was proved to be schwannomas by pathological test. The hearing loss and the facial nerve paralysis (House-Brackmann II) had appeared after operation. NF2 is an autosomal dominant, highly penetrant disease which is characterized by bilateral vestibular schwannomas. Early diagnosis and management for tumor is very important for survival and hearing preservation. The "golden standard" in terms of diagnostic precision is the magnetic resonance imaging (MRI) scan with gadolinium enhancement.
    Lin chuang er bi yan hou ke za zhi = Journal of clinical otorhinolaryngology 09/2006; 20(16):721-3.
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    ABSTRACT: To study the effects of sodium salicylate on the expression of GABAalpha NR1 and hearing response properties of inferior colliculus neurons in mice. Thirty-six kunming mice were divided into three groups (A, B, C,). The expression of GABAalpha NR1 were measured by using RT-PCR. The intensity-rates functions, intensity-latency functions and frequency-turning curves were recorded by extracellular electrophysiological recording techniques. (1) The expression of GABAalpha mRNA of B group was decreased remarkably than the control group (A group, P < 0.05), there weren't noticeable differences between A group and C group (P > 0.05). The expression of NR1 mRNA of B group was increased remarkably than the control group (A group, P < 0.01), there were noticeable differences between A group and C group P < 0.05). (2) The intensity-rates functions, intensity-latency functions were monotonic while the frequency-turning curves were more broad when sodium salicylate was given. (3) The intensity-rates functions, intensity-latency functions were non-monotonic while the frequency-turning curves were sharpened after lidocaine was given. (1) The results suggested that administration of sodium salicylate decreased the expression of GABAalpha while increased the expression of NR1mRNA. (2) The intensity-rates functions, intensity-latency functions were monotonic, the frequency-turning curves were more broad when salicylate was given and the changes above could be reversed by given lidocaine.
    Zhongguo ying yong sheng li xue za zhi = Zhongguo yingyong shenglixue zazhi = Chinese journal of applied physiology 05/2006; 22(2):200-5.
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    ABSTRACT: In order to study the expressions of vascular endothelial growth factor (VEGF) and cyclooxygenase-2 (COX-2) in human laryngeal squamous cell carcinoma (LSCC) and its significance, the expression of VEGF mRNA and COX-2 mRNA in 62 cases of LSCC and 54 adjacent noncancerous laryngeal tissues and 9 normal human laryngeal mucous tissues was detected by using techniques of semi-quantitative RT-PCR. It was found that the expression level of VEGF and COX-2 mRNA was significantly increased in LSCC as compared with that in the normal human laryngeal mucous tissues (both P < 0.01), and the expression level of VEGF and COX-2 mRNA were significantly increased in stage Ill + IV tissues of LSCC as compared with the stage I + II tissues of LSCC (P < 0.01). There was a high positive correlation between VEGF and COX-2 expression in LSCC (r = 0.756, P < 0.01). These data raise the possibility that VEGF and COX-2 may play key roles in the growth, invasion and metastasis of LSCC.
    Journal of Huazhong University of Science and Technology 02/2006; 26(1):105-7. · 0.58 Impact Factor
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    ABSTRACT: To study the expression of perlecan in human laryngeal carcinoma cells and its significance. The expression of perlecan mRNA in human laryngeal carcinoma cells, Hep-2 cells were investigated by using the techniques of semi-quantify RT-PCR. The expression of perlecan in the Hep-2 cells was investigated by using the techniques of immunohistochemistry. It showed that the expression level of perlecan and perlecan mRNA significantly increased in Hep-2 cells as compared with the normal cells, Hacat cells (P < 0.01). These data raise the possibility that perlecan many play key roles in the growth,invasion and metastasis of human laryngeal carcinoma cells through either a paracrine or an autocrine mechanism.
    Lin chuang er bi yan hou ke za zhi = Journal of clinical otorhinolaryngology 01/2006; 20(2):78-80.
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    ABSTRACT: To study the expression of vascular endothelial growth factor (VEGF) and both the Flt-1 and KDR high affinity VEGF receptors in human laryngeal carcinoma cells and its significance. In this study, we investigated the expression of VEGF mRNA and both the Fit-1 mRNA and KDR mRNA high affinity VEGF receptors in human laryngeal carcinoma cells using techniques of semi-quantify RT-PCR. It was showed that the expression level of VEGF mRNA was significantly increased in human laryngeal carcinoma cells as compared with the normal cells, Hacat cells (P <0.05), and there was the high expression level of Flt-1 mRNA in the Hep-2 cells, but not KDR mRNA. These data raise the possibility that VEGF and its receptors many play key roles in the growth, invasion and metastasis of human laryngeal carcinoma cells through either a paracrine or an autocrine mechanism.
    Lin chuang er bi yan hou ke za zhi = Journal of clinical otorhinolaryngology 10/2005; 19(18):842-4.
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    ABSTRACT: The effects of neuroglobin (NGB) gene transfer in vivo mediated by GeneJamer on the hearing response properties of the inferior colliculus (IC) neurons in mice after administration of sodium salicylate were studied. Forty-eight Kunming mice were divided into 4 groups (n=12 in each group): Group A1 (negative control);Group A2 (positive control);Group B, sodium salicylate (450 mg/kg every day) + pEGFP-C1;Group C, sodium salicylate (450 mg/kg every day) + pEGFP-NGB. The GeneJamer and pEGFP-NGB were mixed and injected into IC neurons in mice. The expression of NGB mRNA and protein of IC neurons in mice was detected by using RT-PCR and Western blot methods. The intensity-rate functions, intensity-latency functions and frequency-turning curves in IC neurons were recorded by extracellular electrophysiological recording techniques and the effects of pEGFP-NGB transfer following injection of sodium salicylate on them were studied. It was found that: (1) The GeneJamer-mediated pEGFP-NGB could be effectively transferred into the IC brain tissues in mice and NGB could be expressed intensively. (2) The intensity-rate functions of IC neurons were raised after administration of sodium salicylate. The non-monotonic styles of intensity-rate functions in groups A1, A2 and C were accounted for 74.6%, 72.2 %, 59.3 %, respectively, and the function in group B for 47%. There were significant differences between group B and groups A1, A2 or C (P<0.01, P<0.01, P<0.05). (3) The intensity-latency functions in IC neurons were reduced after administration of sodium salicylate. The non-monotonic styles of intensity-latency functions in groups A1, A2 and C were accounted for 3.2 %, 5.1 %and 21 %, respectively, and that in group B for 45.5 %. There were significant differences between group B and groups A1, A2 or C (P<0.01, P<0.01, P<0.05, respectively). (4) The frequency-turning curves in groups A1 and A2 were sharpened. In 72 acoustic neurons recorded in the group B, the frequency-turning curves from 53 neurons were broadened while those of the rest were sharpened. In group C the frequency-turning curves recorded from 12 of 67 acoustic neurons were broadened while those of the remaining were sharpened. These results suggest that in vivo transfer of NGB gene is highly expressed in IC neurons in mice. In vivo transfer of NGB gene reverses the change of intensity-rate functions, intensity-latency functions and the code styles after administration of sodium salicylate in IC neurons in mice.
    Sheng li xue bao: [Acta physiologica Sinica] 08/2005; 57(4):529-36.
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    ABSTRACT: To observe the glial reactions surrounding facial motor neurons following facial nerve anastomosis. At 1, 7, 21 and 60 d following facial nerve anastomosis, the recovery process of facial movement was observed, the glial fibrillary acidic protein (GFAP) immunoreactivity was analyzed by a combined method of fluorescent retrograde tracing and immunofluorescent histochemical staining, and the ultrastructure of astrocytes were observed under a transmission electron microscope (TEM), respectively. Postoperatively the function of facial muscles could not return to normal, often accompanied with hyperkinetic syndromes such as synkinesis at the late stage. Motor neurons in every facial subnucleus could be retrogradely labeled by fluoro-gold (FG), and displayed an evident somatotopic organization. Normally there was a considerable number of GFAP-positive cells in nonnucleus regions but few inside the facial nucleus region. Postoperatively the GFAP immunoreactivity in the anastomotic side increased significantly, but gradually decreased at the late stage. The ultrastructure of astrocytes in our experiment showed that the sheet-like process of astrocytes invested and protected the injured facial motor neurons. The present study shows that reactive astrocytes undergo some characteristic changes during the process of facial nerve injury and regeneration. The plastic change at the late stage may be involved in the mechanism of synkinesis.
    Journal of Huazhong University of Science and Technology 02/2005; 25(6):726-8. · 0.58 Impact Factor
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    ABSTRACT: To investigate whether apoptosis is one of the mechanism in the immune response of inner ear,and to detect the expression of Fas, FasL, Bcl-2 and Bax in the inner ears. Sixteen healthy, female guinea pigs were employed in the experiment. Sensitized systematically with keyhole limpet hemocyanin (KLH), the KLH-immunized animals were inoculated with the same antigen, and the control animals were injected PBS through cochlea basal turn. The animals were sacrificed at 7 day after inner ear vaccination. Transmission electron microscopy was used to detect inner ear apoptotic cells, and paraffin sections of cochlea from animals were stained using a terminal-deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL) assay to identify inner ear cells undergoing apoptosis. Immunohistochemistry method was used to detect the expression of Fas, FasL, Bcl-2 and Bax in the inner ears. The observation of electron microscopy had shown the features characteristic of apoptotic cells in the KLH-immunized inner ears but not in the control inner ears. TUNEL-positive cells were found in the KLH-immunized inner ears but not in the control inner ears. The positive cells were the hair cells in Corti's organ, and the marginal cells in the stria vascularis and the neurons in the spiral ganglion. Moreover under morphological analysis by light microscope, these cells had the features characteristic of apoptosis. High expression of Fas and FasL could be detected in Corti's organ, the stria vascularis, the spiral ligament and the neurons of the spiral ganglion in the KLH-immunized inner ears. A low expression of Fas could be detected in the stria vascularis and the neurons of the spiral ganglion in the control inner ears, but no cells staining positive for FasL were found in the control inner ears. No cells staining positive for Bcl-2 were found in the KLH-immunized animals but moderate expression of Bcl-2 could be detected in Corti's organ, the lateral wall and the neurons of the spiral ganglion in the control inner ears. High expression of Bax could be detected in Corti's organ, the lateral wall and the neurons of the spiral ganglion in the KLH-immunized inner ears. A low expression of Bax could be detected in the neurons of the spiral ganglion and no cells staining positive for Bax were found in Corti's organ, the lateral wall in the control inner ears. These findings suggest apoptosis is involved in the pathogenesis of the immune response of inner ear and Fas- FasL pathway is one of important signal transportation of the course and Bcl-2 and Bax have a critical role in the regulation of apoptotic cell death induced by the immune response of inner ear.
    Zhonghua er bi yan hou ke za zhi 12/2004; 39(11):663-8.
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    ABSTRACT: To observe the effect of uPA on nerve regeneration and the expression of fibronectin and laminin after the facial nerve were transected and anastomosed, and to explore the mechanism of uPA in the microenvironment of nerve regeneration. Twenty four SD rats were randomly divided into two groups: uPA group and control group. Every rat's extracranial facial were transected and anastomosed. Then uPA were injected at the belly of rat subcutaneously for five days in uPA group, three times one day, 5000 U/kg one time. Equal volume saline were injected in control models at the same site and the same time. At 1, 7, 21, 42, 84 day of post-operation, the diameter, perimeter, myelin sheath area of fibers and the expression of fibronectin and laminin in early phase of post-operation were measured by using morphometry, immunochemistry and the image analysis. After operation, myelin sheath area at 21, 42 day and diameter of new regenerated fiber at 42 day in uPA group were better than those in control group (P<0.05). The expression of fibronectin elevated gradually after operation, and it expressed more than that of control group at 14 and 21 day (P<0.05). The expression of laminin decreased after operation, then elevated with the progress of nerve regeneration and showed no significant difference between two groups. The uPA can promote the fibrin degradation and upregulate the fibronectin expression when applicating in rat model, which benefit to improve the microenvironment and enhance facial nerve regeneration.
    Lin chuang er bi yan hou ke za zhi = Journal of clinical otorhinolaryngology 11/2004; 18(10):623-6.

Publication Stats

16 Citations
4.69 Total Impact Points

Institutions

  • 2011
    • Capital Medical University
      • Department of Otorhinolaryngology Head and Neck Surgery
      Peping, Beijing, China
  • 2004–2010
    • Huazhong University of Science and Technology
      • Department of Otorhinolaryngology, Head and Neck Surgery
      Wu-han-shih, Hubei, China