Objective To study the changes of N-methyl-D-aspartate receptors(NMDARs) expression at local sy-(napses) in auditory cortices after ototoxic deafened and following electrical intracochear stimulation(EIS).Methods We prepared highly purified synaptosomes from primary auditory cortex by two discontinuous gradient centrifugations, and compared the differences of NMDA receptors expression in ototoxic deafened group control group, normal control group and different experimental groups (EIS from 0.5h to 2h ) by Western blotting.Results We found that EIS for as little as half and one hour the adult and developmental rats of auditory deprivation can induce a significant increase in NMDAR subunit2A (NR2A) protein. But the changes of NMDAR subunit1(NR1) and NMDAR subunit2B(NR2B) are not significant.Conclusion There is a rapid activity-dependent expression of synaptic NR2A receptors in primary auditory cortex in vivo.

The association of viral infection to ear disease has triggered a great deal of interests. In the present paper, we provide a critical review of the viral hypothesis of ear diseases. Detection of viral antigen and antibody or RNA and DNA in the patients serum, endolymphatic fluid or surgical pathology specimens reveals that virus may have relevance to certain kinds of ear diseases, such as Meniere's disease, idiopathic sudden sensorineural hearing loss, otosclerosis. Bell's palsy and otitis media. The most appealing is the herpesvirus, which can cause latent infection in the neurons, and its reactivation may be the mechanism of recurrent attacks of ear diseases. Currently, antiviral drug treatment plus supportive therapy are the most effective managements dealing with viral infection. Although antiviral vaccine will become a promising preventive strategy in the future.
Ear Diseases ; virology ; Humans ; Labyrinth Diseases ; virology ; Otitis Media ; virology ; Virus Diseases

OBJECTIVE To explore whether auditory event-related potential (AERP)exists in rat and what its features are,and to set up a set of AERP recording technique. METHODS Ten normal juvenile and 10 adult rats were served as subjects. Each rat was anesthetized and recorded AERP with passive mode. The responsibility of AERPs between juvenile and adult rats was compared. Also,the responsibility of AERP between rats and normal humans was compared. RESULTS AERPs could be evoked with passive recording mode in rats. Although the latencies and amplitudes of juvenile rats were respectively shorter and lower than thoses of adult rats,there were no significance in statistics. The AERP waveform of rats was similar to that of humans essentially. However,compared to the humans,the responsibility and stability of AERP in rats were less satisfied. CONCLUSION A similar AERP exists in rat and human. AERP has a process of development,differentiation and maturation,which may accomplish in early age in rat. Rat can serve as the candidate animal for AERP research.

Objective To study the neuroelectrophysiological responsibility, mechanism and clinical application of auditory event-related potential (AERP) generated in bilateral auditory centers with ipsilateral and contralateral recordings. Methods 14 normal young adults were served as subjects. AERPs, including the responsibility, latencies, amplitudes and waveforms, were simultaneously recorded on ipsilateral and contralateral vertexes and analyzed. Results The latencies and amplitudes of AERPs did not show significicant difference statislically between ipsilateral and contralateral vertexes recordings. However, the responsibility of AERP recorded on ipsilateral vertex was more satisfied than that on contralateral vertex, which exhibited smoothing waves, discriminative wave peaks and little hetero-wave. Conclusion The origins of AERP in bilateral auditory centers were essentially symmetrical. Nevertheless, the responsibility of AERP recorded on ipsilateral vertex was more satisfied than that on contralateral vertex, which was related to the components of auditory subcortex, transmission of auditory nerve to bilateral centers and the contralateral inhibiting effect of medial olivo-cochlear system. The effects of multi-generation and multi-component of AERPs on the clinical application should be considered.

Objective To study the immunogenicity of Streptococcus pneumoniae pneumolysin DNA vaccine in Rhesus macaques. Methods The deletion of the gene sequence encoding for the 11 amino acids at the carboxyl terminal of pneumolysin (PN) from its wild type gene (pn) by PCR resulted in a mu-tant pneumolysin gene (pnd). The wild type pn gene encoding PN and the mutant gene (pnd) encoding PND were cloned into pVAX1 vector respectively and then tested as Ppn and Ppnd DNA vaccines. The PN and PND proteins were purified from recombinant E. coli. Rhesus macaques were immunized by intramuscu-larly (i.m.) injection of Ppn or Ppnd DNA vaccine with electroporation (EP). Results Because of the deletion of the gene sequence encoding for the eleven amino acids at the carboxyl terminal of the PN from pn gene, the recombinant PND antigen lost its hemolytic activity while its antigenicity was remained. The spe-cific humoral immunity against pneumolysin was induced by injecting monkey with 500 μg DNA followed by EP. Boosting the Ppn or Ppnd DNA/EP primed animals with corresponding recombinant protein, PN or PND, evoked strong immune response at about 4 fold increase in the antibody titer. Conclusion Specific antibody responses were induced in the monkeys by DNA vaccination and electroporation. The immunogenic-ity of the DNA vaccines were significantly enhanced when PN or PND protein boost was applied 10 d after third DNA vaccination.

Objective To investigate the relationship between 5 ,10-methylenetetrahydrofolate reductase (MTHFR) gene C677T locus polymorphism and unexplained adverse pregnancy.Methods Three hundreds and twenty pregnant women with unknown ad-verse pregnancy ≥ 2 times in the infertility department ,and obstetrics and gynecology department of our hospital from June 2014 to May 2016 were selected as the case group and 388 healthy non-abortion parous women were taken as the control group.The MTH-FR gene C677T locus polymorphism in the two groups was analyzed by using gene chip method.The differences in genotype and al-lele frequency distribution were compared between the two groups ,and the relationship between unexplained adverse pregnancy and MTHFR C677T locus polymorphism was analyzed.Results The frequency distribution of MTHFR genotype C677T C/C had sta-tistically significant difference between the case group and control group (P<0.05 ,OR=0.284).The frequency distribution of gen-otype C/T had no statistical difference between the case group and control group (P=0.400 ,OR=1.140).The frequency distribu-tion of genotype T/T had statistical difference between the case group and control group (P<0.05 ,OR=7.672).The frequency distribution of allele C and T had statistical difference between the case group and control group ( P< 0.05 ,OR= 0.304 ). Conclusion The high expression of MTHFR C677T genotype T/T may be a risk factor of unknown adverse pregnancy in child-bearing age women.

Objective To explore the imaging diagnostic method of Merkel cell carcinoma(MCC). Methods Imaging and pathologic features of MCC of 1 female and 44 years old patient were determinded by CT, MRI, pathologic examination and operative observation. Results MCC is a rare subepidermal carcinoma, and most located in the head, neck region and extrimities. Clinically, only a presumptive diagnosis of MCC can be established. The definite diagnosis of MCC can be made only by pathologic examination, especially immunohistological method. MCC is red and brown, and looks like rotten meat. MCC texture is middle with rich blood supply. MCC can invade occipital bone, occipital muscles, dura and cerebellar tissues. Optical microscope observation showed that MCC is located at dermis, and sometimes at subcutaneous tissues. MCC cells are gathered in groups, their sizes are unanimous, and their shapes look like lymphocytes. The cell nuclei of MCC looklike empty bubbles, and have many karyokinesis. The chromatin of MCC is very fine. The cytoplasms of MCC are little, and oxyphil staining. Immunohistological staining showed that both NSE (neuron-specific enolase) and CK(cytokeratin) are positive. The CT imaging of MCC is higher density. The MRI imaging of MCC is lower signals with unclear border in T1w and T2w, but becomes clear in enhanced MRI. Tumor cells invasion to soft tissues and bones could be seen on CT or MRI. Conclusion CT and MRI could clearly show the characteristics of MCC, and provide valuable information for operative treatment of MCC.

PurposeTo evaluate the curative effect and possibility of the intraarterial chemotherapy followed by radical hysterectomy for locally advanced cervical cancer. Methods59 patients with FIGO stage lb-IV cervical cancer were enrolled and the tumor size with stage lb and Ua must be more than 4 cm. Treatment consisted of bilateral internal iliac artery infusion of cisplatin 80 rog, 5-Fu 1 500 rog, and AT1258 60 mg for 1 - 3 courses separated by 3 weeks. The tumor size was measured by vagina ultrasonic and CT before and after chemotherapy to calculate the shrunk size percent. All patients underwent radical hysterectomy. Patients with disease in the vagina, parametrium, pelvic lymphnodes or microscopic tumor emboli were followed postoperative pelvic irradiation. Results59 patients underwent 122 courses of intraarterial chemotherapy for 1 - 3 courses each. The total response rate was 95 %. Complete response (CR) was achieved in 18 of 59 patients (31%) ,while a partial response (PR) was noted in 38 (64%) and stable disease (SD) in 3 (5% ). No patient had progressive disease. 14 patients with pelvic lymphnode metastasis were all found to have PR and SD. No patient had lymphnode metastasis in CR. Of 18 patients with CR, 17 patients had not recurrence. Only one patient with stage lla recurred 20 months after operation. But he also cured. All 18 patients with CR were living free of disease. But 3 in 38 patients with PR died and 3 patients with SD were all died. There were no severe complications related to intraarterial chemotherapy and radical hysterectomy. Conclusionslntraarterial chemotherapy before operation can shrink tumor size, decrease the lymphnode metastasis. It is feasible in helping elevating cure rate of patients with locally advanced cervical cancer who are at high risk for failure with conventional treatment.

Objective To explore the value of DWI combined with dynamic contrast-enhanced MR imaging (DCE MRI) in differential diagnosis of benign and malignant parotid gland tumors.Methods DWI and DCE-MRI was performed in 90 patients with parotid gland tumors (94 lesions),included 75 benign tumors and 19 malignant tumors.The ADCs were measured,and the type of time-intensity curve (TIC) was assessed.ROC curve was used to analyze the diagnostic efficacy of ADC values,TIC types and combination of the two methods in diagnosing benign and malignant tumors.Results The ADC values of malignant tumors ([1.02 ± 0.25] × 10a mm2/s) were significantly smaller than that of benign tumors ([1.38±0.44] × 10-3 mm2/s;t=5.170,P＜0.001).The sensitivity was 0.62 and specificity was 0.92,when type A (persistent),B (washout),and D (flat) tumors regarded as benign and type C (plateau) tumors regarded as malignant.Sensitivity and specificity were significantly improved in diagnosis of malignant tumors when combined type C and ADC＜1.36× 10-3mm2/s,which were 0.92 and 0.95,respectively.Conclusion Due to an overlap within benign and malignant parotid tumors,the diagnosis should not be addressed on the basis of ADC values alone.The TIC manifestations type A,B or D on DCE-MRI indicates benign parotid tumors,and type C indicates malignant diseases.When the TIC shows type C,combining ADC value can elevate the differential diagnostic efficacy of benign and malignant parotid tumors.

Objective To prepare the chitosan-potD nanoparticles and to evaluate its protective efficacy against pneumococcal nasopharyngeal colonization. Methods potD gene was amplificated from pneumococcal genome and was inserted into pVAX1 expression vectors to construct pVAX1-potD recombinant plasmid which was then transfected into 293T cell using LipofectAMINE 2000 to analyze transient potD gene expression in vitro by RT-PCR and Western blot. Chitosan-potD nanoparticles were freshly prepared by coacervation methods at each time and the characterizations of the nanoparticles were then evaluated. BALB/c mice were immunized with chitosan-potD, naked potD DNA or pVAX1 for 4 times at two-week intervals. Anti-PotD IgG, IgG1 and IgG2a levels in serum and IgA levels in nasal washes, bronchoalveolar lavage fluids (BALF) and middle ear lavages(MEL) were detected by indirect enzyme-linked immunosorbent assay (ELISA). IL-17A, IL-4 and IFN-γ levels in splenocytes were determined by double sandwich ELISA. Mice were intrannsally challenged with Streptococcus pneumoniae ATCC6303, and Pneumococci were recovered from the nasopharyngeal niche at the fifth day after challenge. Results potD gene was successfully amplificated by PCR and the sequence was confimed to be consistent with that in the Genbank. The pVAX1-potD recombinant plasmid was successfully constructed and was expressed in eukaryocytes in vitro. The mean size and zeta potential of chitosan-potD nanoparticles was 430 nm and + 20.5 mv, respectively. Chitosan-potD nanoparticles were not digested by DNase Ⅰ , while naked potD DNA was completely digested. The levels of antibodies inculding IgG, IgG1, IgG2a, IgA and cytokines including IL-17A, IL-4 and IFN-γ were significantly higher in mice immunized with chitosan-potD nanoparticles than mice with naked potD or pVAX1 ( P <0.05) only. More importantly, much less Pneumococci were recovered from mice immunized with chitosan-potD nanoparticles than the other groups(P <0.05). Conclusion Chitosan-potD nanoparticles significantly enhanced the immunogenicity and protection efficacy of DNA vaccines by intranasal immunization and could be used as a potential mucosal vaccine to prevent pneumococcal infection.