Objective The aim of the study was to analyse the middle ear function of one hundred and twenty-five newborns using 226,678,1 000 Hz tympanometry. Methods Tympanometries with 226,678,1 000 Hz probe tones were obtained from 125 newborns (250 ears) by means of GSI Tympstar middle ear analyzer. The newborns with abnormal tympanograms were referred for temporale CT. Results One case of 226 Hz tympanograms was type "As" while others were all type "A",of which single-peaked tympanograms was 43.2%(108/250),W-shaped tympanograms was 56.8%(142/250). The abnormal pattern of 678 Hz tympanograms were 74 ears,and the abnormal pattern of 1 000 Hz tympanograms were 35 ears. 43 newborns (86 ears) have accepted temporale CT and results showed 30 middle ears with effusion. Conclusion 226 Hz probe tone tympanometry couldn't reflect the middle ear function of newborns. The sensitivity of 678 Hz probe tone tympanometry in the diagnosis of middle ear function of newborns was good,but specificity was worse. The sensitivity and specificity of 1 000 Hz probe tone tympanometry were the best.

Objective To study the effects of multi frequency tympanometry and acoustic stapedius reflexes on the diagnosis of the middle ear functions of infants.Methods Tympanometries with 226,678 and 1 000 Hz probe tones and 500 and 1 000 Hz acoustic stapedius reflexes were performed on infants at age 0~1 year.128 normal infants(256 ears) and 104 abnormal infants(168 ears) were tested with GSI Tympstar middle ear analyzer.Results The patterns of 226 Hz tympanograms were non-peaked,single-peaked and W-shaped.There were no significant differences between two groups of infants aged 0～6 months.The patterns of 678 Hz and 1 000 Hz tympanograms showed significant differences between two groups of infants aged 0～1 year.The outcomes of 500 Hz and 1 000 Hz acoustic stapedius reflexes showed no differences between the same age groups.There were significant differences between normal group and abnormal group.Conclusion Tympanometry with 678 Hz and 1 000 Hz probe tones can yield accurate diagnostic results in the assessment of the middle ear functions for infants younger than six months.The tympanometry with three frequency probe tones is sensitive to infants aged 6～12 months.The outcomes of acoustic stapedius reflexes at 500 and 1 000 Hz show no differences between the same age groups,but presences become more noticeable as a function of ages.

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 screen the optimal formula and technology of demethoxycurcumin nano-structured lipid carriers. Meth-ods:The encapsulation efficiency as the main investigation index, the single factor exploration and orthogonal design were used to study the main factors affecting the quality of the nanoparticles. The optimal formula and technology were obtained. Results:The optimized parameters were as follows:the ratio of drug to lipid materials was 1 ∶40, the ratio of liquid lipid to total lipid materials was 10%, the amount of surfactants was 4% and the amount of lecithin was 2%. The prepared nanoparticles were spheric and regular. The size dis-tribution of the nanoparticles was narrow with the average particle size of 110nm and PDI of 0. 199. Conclusion:The optimized formula and technology of demethoxycurcumin nano-structured lipid carriers are stable and practicable,which provide reference for the further research of demethoxycurcumin.

From May, 2000 to June, 2001, 27 patients with Parkinson disease (PD), including 10 cases of rigidity, 13 cases of tremor, 4 cases of rigidity and tremor, were treated by microelectrode-guided technique. Among them, phlebotomy was carried out in 17 cases and thalamotomy in 10 cases. All the targets of lesion were anatomically located by using MR and neurophysiological signals on microelectrode. Our results showed that the efficiency of microelectrode-guided technique for treatment of PD was 98%. The postoperative unified parkinson disease rating scale were 12.3 +/- 9.1 and 13.2 +/- 8.9 respectively, which significantly improved as compared with those before operation. It was concluded that by recognizing special electrical signals in neurons microelectrode-guided neuropsychological techniques can locate target at cellular level, which overcomes the individual difference in anatomy and function, and allow more accuracy, safety and efficiency of operation. This is especially true of PD patients who fail to respond to medical treatment.
Magnetic Resonance Imaging ; *Microelectrodes ; Parkinson Disease/*surgery ; Stereotaxic Techniques ; Thalamus/surgery

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.

Objective To explore the influence of minimally invasive puncture drainage on blood brain barrier (BBB) function and its mechanism.Methods Ninety-two patients with hypertensive intra-cerebral hemorrhage (HICH) in the Department of Neurosurgery of Jiaxing Affiliated Second Hospital of Zhejiang Province were divided into a control group and an observation group, according to random number table method, 46 cases in each group. In the control group, the conventional craniotomy was performed, while in the observation group, minimally invasive puncture drainage was carried out to remove the hematoma. The National Institute of Health Stroke Scale (NIHSS) were used to evaluate the neural function, the level of serum myelin basic protein (MBP) was detected by enzyme linked immunosorbent assay (ELISA), the central nervous specific serum protein S100 level was measured by electrochemical luminescence method, the albumin levels in serum and cerebrospinal fluid were determined by automatic biological analyzer, and the BBB index was calculated. After 14 days of surgery, the curative effect and incidence of complications of two groups were observed.Results After surgery, the NIHSS scores of two groups were obviously lower than those before surgery, and the degree of descent in observation group was more significant than that in the control group (score: 3.68±2.39 vs. 5.43±3.89,P < 0.05); after surgery, the levels of MBP, S100, albumin in cerebrospinal fluid and BBB in two groups were higher than those before surgery [MBP (μg/L): 3.02±0.28 vs. 3.81±0.29, S100 (μg/L): 0.95±0.24 vs. 1.34±0.27, cerebrospinal fluid albumin (μg/L): 9.89±0.78 vs. 21.43±1.14, BBB index: 0.22±0.04 vs. 0.48±0.05], the differences being statistically significant (allP < 0.05), but the change values in the observation group were less significant than those in the control group. The total effective rate in observation group was significantly higher than that in the control group [84.78% (39/46) vs. 65.22% (30/46),χ2 = 4.696,P = 0.030]. The incidence of wound infection, gastrointestinal bleeding in observation group was markedly lower than that in the control group [16.67% (6/46) vs. 36.96% (17/46), χ2 = 4.120,P = 0.042].Conclusion The minimally invasive puncture drainage has unequivocal clinical curative effect in treatment of patients with HICH, it can protect the nerve and BBB functions and reduce the incidence of complications.

OBJECTIVE To prepare pneumolysin(Pn)by genetic engineering and thereby establish the basis for the study of vaccines against otitis media. METHODS A pair of primers including two restriction sites was designed based on the pneumolysin gene sequence reported by Walker in 1987. The pneumolysin gene was PCR-amplified from pneumococcal DNA. The resulting fragment, digested by restriction enzymes, was ligated into the vector PET-28a and then transformed into host cell E.coli JM109(DE3). RESULTS The sequence of the inserted pneumolysin gene was confirmed by DNA sequencing. CONCLUSION The pneumolysin gene was successfully cloned into the host cell.

Objective To study the clinical value of air-and hone-conduction auditory brainstem responses(ABRs) in children with congenital aural atresia.Methods Air-and bone-conduction clickevoked ABRs in 38 children with congenital external auditory canal atresia were compared to those in 34 normal-hearing children.Results ABR threshold were (65.0 ± 8.7) dBnHL for air conduction and (10.6 ±7.0) dBnHL for bone conduction in children with congenital external auditory canal atresia;and (25.6 ± 1.7) dBnHL and (9.7 ± 6.7) dBnHL respectively in normal hearing children.There was statistically difference between the two groups in air-conduction ABR threshold(t' =21.59,P ＜ 0.01).There was no significance between the two groups in bone-conduction ABR (P ＞ 0.05).While the air-bone ABR threshold gap was greater in children with external auditory canal atresia than in normal hearing children (t' =13.78,P ＜ 0.01),bone-conduction ABR wave latencies were not statistically different between the two groups(all P ＞ 0.05).Conclusion Bone conduction ABR is valuable in assessing fuuction of cochlea,auditory nerve and brainstem in individuals with congenital external auditory canal atresia.It has important clinical value in objective differential diagnosis of conductive deafness with combined application of air-and bone-conduction ABRs.

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.