BACKGROUND: Although cultured technique and purified method of Schwann cells (SCs) are changed successively,purity is still limited. Whether double negative selections can culture high-purified SCs still needs a further study.OBJECTIVE: To investigate the effects of double negative selections on growth and purity of SCs.DESIGN: Controlled animal study.SETTING: Department of Otolaryngology Head and Neck Surgery, Beijing Tongren Hospital, Capital Medical University,and the Beijing Institution of Otolaryngology.MATERIALS: The experiment was carried out in the Department of Otolaryngology Head and Neck Surgery, Beijing Tongren Hospital, Capital Medical University and the Beijing Institution of Otolaryngology between May 2006 and July 2006. Twenty new-born Wistar rats of 3 days (without gender limitation) were provided by the Experimental Animal Center of the Chinese Medical Academy.METHODS: The sciatic nerves of rats were digested by 0.2% collagenase. ① Grouping: All the rats were divided into 4 groups with 5 each. The experimental group was depurated by differential adhesion and Ara-c for 30 minutes. SCs in the control group 1 and 2 were managed by differential adhesion and Ara-c for 30 minutes, respectively. Cells in the control group 3 were not purified. ② Evaluation: The cell growth was observed under invert microscope; growth curves were measured with MTT method in the experimental group and control group 3 to observe cell proliferation. In addition, the SCs were identified by immunocytochemistry; the culture purity was calculated (ratio of S100 positive cells in each sight).MAIN OUTCOME MEASURES: ① Growth of SCs; ② immunocytochemistry stain of SCs; ③ growth curve of SCs; ④ purity of SCs.RESULTS: ① Growth of SCs: At 5 days after incubation, the SCs proliferated well. SCs were spindle, bipolar and sometimes tripolar, with obvious nuclei and a little cytoplasm. There was more fibroblast cell in the group 3 than that in experimental group. ② Growth curve: The SCs in experimental group entered into the logarithm multiplication period at the third day. ③ Immunocytochemistry: The positive-stained SCs were brown in its body and process, and arranged like swirl. The FB with spread and anomaly shape was slight blue. ④ The ratio of anti-S100 positive cells was higher in experimental group than that in group 3 (P ＜ 0.01).CONCLUSION: With the double negative selection, high-purified SCs are obtained effectively.

Objective To know the nursing students' willingness engaging in geriatric nursing and analyze affecting factors so as to provide references for nurturing professionals for caring for the aged.Methods 231 nursing students were surveyed.The questionnaire included general information,life experience with the elderly,willingness engaging in geriatric nursing,attitude towards the elderly and cognition of aging.The investigation results underwent analysis.Results Single factor analysis showed that whether ever living with the elderly (x2=6.027),time living with the elderly and relationship with the elderly (x2=4.928) influeced the nursing students' willingness engaging in geriatric nursing.Multiple correspondence analysis indicated that nursing students living with elderly more than one year and having good relationship with the elderly had positive willingness.Conclusions Geriatric nursing curriculum should be optimized and more opportunities should be created for nursing students to be in touch with the elderly.At the same time,we must pay attention to occupation guide and moral education about how to respect and care for the elderly.

Objective To explore the establishment and evaluation of various forms of complementary transfer. Methods The former method using the traditional handover method, the establishment of various forms of complementary shift model improved, including optimized bedside handover, handover of patients′ bedsore image and emotional barometer over emotional changes, establish WeChat group to observe improvement;the first two months (99 cases, monitoring 42 days) and the improvement after the six months (97 cases, monitoring 43 days) score of nurses for the patient to grasp, bedside shift time and transfer the missing information. Concrete research and implementation methods, research objects, sample size, measurement indicators, evaluation methods, etc. Results Both shift mode, there is no information on patients admitted to general statistical difference (P>0.05);under complementary shift mode, nurse the patients′ condition from the master score (80.95 ± 4.30) points increased to (84.88 ± 4.01) points, in which the patients′condition data mastery score from (40.81 ± 2.02) points increased to (44.16 ± 2.88) points, nursing problems and measures of control points value from (13.21 ± 1.54) points increased to (14.44 ± 1.72) points, the difference was statistically significant (t =4.362, 6.209, 3.462, P<0.01);general information master score, treatment information at its disposal no difference scores statistically significant (P>0.05). Bedside nurse shift change time decreased from former improvements (31.21 ± 2.63) minutes to (14.42 ± 2.40) minutes, and the difference was statistically significant (t=30.767, P=0.000). Shift nurse missing information dropped from 16 times to 7 times, the difference was statistically significant (χ2=5.124, P=0.024). Conclusions Complementary model can reduce the insufficiency of current clinical succession work, help nurse master the patients′physical and mental conditions and more comprehensive understanding of the ward, reduce the omission or missing information and improve the working efficiency and responsibility.

Based on microfabrication technology and electrochemical modification method, a micro electrochemical sensor for nitrate ( NO-3 ) determination was developed. A micro sensor chip with working electrode and counter electrode was used as the signal convertor of the sensor. The area of the micro working_electrode was only 1 mm2 . As an electrocatalysis sensitive material, copper was electrodeposited onto the working electrode by square_wave pulse current electrodeposition method. The morphologies and components of freshly deposited materials were examined by scanning electron microscopy ( SEM ) and X_ray diffraction ( XRD) to explore key factors that affected the electrocatalytic ability of the deposited copper layer for reducing nitrate ions. The experimental results revealed that under the optimal conditions, the deposited copper layer was macroporous and had a larger effective surface area that could serve as a more effective electrocatalyst in facilitating nitrate reduction. Electrochemical response of the macroporous copper layer was characterized by linear sweep voltammetry in acidic supporting electrolytes ( pH=2 ) . The electroanalytical results showed that the modified microsensor had marked sensitivity for standard nitrate samples within the concentration range from 12. 5 to 3000 μmol/L (in the range of 12. 5-200 μmol/L yielded straight line:y1=-0. 1422x-10. 326, R12=0. 9976, while in the range of 200-3000 μmol/L yielded straight line: y2=-0. 0984x-22. 144, R22=0. 9927) with a detection limit of 2 μmol/L (S/N=3). The developed electrochemical microsensor was also employed for nitrate determination in water samples collected from lakes and rivers near the city of Beijing. The results were in good agreement with the data given by qualified water quality detection institute, with the deviations from 3 . 9% to 15 . 4%.

Objective To compare the effects of different administration routes and dosages of tropisetron on cisplatin-induced kaolin intake in rats.Methods Ninety-six healthy adult male Wistar SPF rats were randomly divided into 8 groups(n =12 each):intrathecal (IT) control group (group TC) and 3 tropisetron groups receiving IT tropisetron 10,20 and 30 μg,the volume of each group was 30 μl (group T10,T20,T30),intravenous(Ⅳ) control group (group IC) and 3 tropisetron groups receiving Ⅳ tropisetron 0.3,0.5 and 0.7 mg/kg respectively (group I0.3,I0.5,I0.7).In group TC and IC,normal saline 30 μl and 0.5 ml were injected IT and Ⅳ,respectively.All rats received cisplatin 3mg/kg by intraperitoneal injection at the time point of thirty minutes after administration,each rat weight,the daily food and kaolin intakes were detected at the time point of 48 hours after cisplatin administration.Results Compared with group Tc,each rat weight loss,the kaolin intakes were significantly decreased (P ＜ 0.05),and food intake dose was significantly increased in group T20 (P ＜ 0.05).Compared with group IC,each rat weight loss,the kaolin intakes were significantly decreased (P ＜ 0.05),and food intake dose was significantly increased in group I0.5 and I0.7 (P ＜0.05).There was no significant difference between group I0.5,I0.7 and group T20.Conclusions The kaolin intakes and the rat weight loss can be decreased by IT tropisetron,and the food take dose was increased meanwhile,and IT tropisetron 20 μg has equivalent efficacy to IV tropisetron 0.5 or 0.7 mg/kg.IT could be the new administration route of tropisetron.

Objective To investigate the effect of small interfering RNA (siRNA) suppressing discoidin domain receptor 1 (DDR1) gene on biological behaviour of Kupffer cells (KC) in acute hepatic injury. Methods Male BALB/c mice were randomly divided into control, hepatic injury model, non-silencing siRNA and DDRlsiRNA groups. Hepatic injury model induced by intravenous injection of Con-canavalinA (ConA) 15 mg/kg, with or without hydrodynamic tail intravenous injection of naked siRNA (50 μg,2.0-2.5 mg/kg)/mouse or 1.5 ml normal saline. The expression of DDRI was assayed by West-ern blot and pro-inflammatory cytokine expression analyzed by ELISA. In the meantime, alanine amin-otransferase (ALT) and Kupffer cells'clearance to carbon granules was detected. Results The expres-sion of DDR1 obviously increasod at 6 h after hepatic injury, roached peak at 24 h and began to decrease at 48 h. Pretreatment with DDRisiRNA could obviously inhibit the expression of DDR1 and abrogate the high levels of ALT, expressions of TNF-α and IL-1β as well as phagocytosis of Kupffer cells. Conclu-sions Inhibition of discoidin domain receptor 1 by in vivo delivery of siRNA attenuates ConA-induced hepatic injury. Possible mechanism is that the inhibition of activity of KC inhibits the expression of pro-in-flammatory cytokines and thus alleviates hepatic injury.

OBJECTIVE The aim of this study is to investigate the correlation between the spontaneous otoacoustic emission(SOAE)and transient evoked otoacoustic emission(TEOAE)in neonate. METHODS The subjects were 224 ears of 112 newborns(59 females, 53 males). The age of subjects ranged from 2 to 4 days (mean?SD: 2.68?0.74 day years). SOAE and TEOAE were measured with ILO96. RESULTS The correlation between SOAE number and TEOAE level were found (r=0.43, P

Objective To investigate the protective effect of CBLB 502 on radiation pneumonitis and pulmonary fibrosis for confirming the feasibility of CBLB502 as a clinical anti-radiation drug release.Methods With a single 20 Gy irradia-tion, C57BL/6J mice was sacrificed on 24 h, 1 month, 3 months and 5 months and lung tissue was assayed by TUNEL method for apoptosis of alveolar epithelial cells and endothelial cells , HE staining showing fibrosis changes , immunohisto-chemistry detecting the expression of specific indicators , as well as pathological changes of the fur and skin radiated site . Results CBLB502 inhibits apoptosis in mice alveolar epithelial cells and vascular endothelial cells after irradiation , slowing the process of pulmonary fibrosis , while reducing the expression of laminin and maintaining the expression of surfac-tant protein B, and the skin inflammation also significantly reduced .Conclusion CBLB502 could alleviate the occurrence of radiation pneumonitis and pulmonary fibrosis as well as radiation-induced skin injury .

Human ribosomal DNA (hrDNA) targeting vector pHr is a homologous recombinant plasmid for human genome which developed in the State Key Laboratory of Medical Genetics. pHr was used to construct a recombinant plasmid pHr-CMG expressing mda-7/GFP fusion gene and its efficacy in the hepatocellular carcinoma cell line Bel-7402 was investigated. The expression of mda-7/GFP fusion gene was detected by fluorescent microscope, RT-PCR and Western blotting, and its function was detected by cell-cycle analyses, MTT assay and Hoechst33258 staining. The results demonstrated that pHr-CMG vector could express MDA-7/GFP fusion protein effectually and the mda-7 gene could induce cell apoptosis and proliferation suppression in Bel-7402 cell line, which might be caused by the G2/M cell cycle arrest. These results also suggested that human ribosomal DNA targeting vector system and the pHr-CMG vector may be applied in further gene therapy researches for hepatocellular carcinoma.

Objective:To explore the putative role of CD40 and COX-2 expression in human esophageal squamous cell carcinoma(ESCC)and to analyze their possible relationship with angiogenesis in ESCC.Methods:The expression of CD40 and COX-2 was detected in 79 ESCC and 28 normal esophageal epithelial tissue samples with immunohistochemical staining.The microvessel density by CD34 was determined and the clinicopathological significance of CD40 and COX-2 expression in ESCC was analyzed.In addition,the expression of CD40 and COX-2 in esophageal squamous cell carcinoma cell line Eca109 and primary cultured normal esophageal epithelial cells in vivo was comparatively studied with immunocytochemical staining andWestern blot.Results:Compared with that in normal epithelial tissues,the expression of CD40 and COX-2 in ESCC was significantly higher(54.43%vs 10.71%:69.62%vs 17.86%:respectively,P<0.05).The positive expression rate of CD40 in ESCC cases with lymph node metastasis was significantly higher than that in those without lymph node metastasis(70.37%vs 46.1 5%.P<0.05).No correlation was found between CD40 expression and patient age,sex,tumor location,size and differentiation of tumors.No clinicopathological significanca of COX-2 expression in ESCC was found.There was a positive correlation between CD40 expression and COX-2 expression in esophageal squamous cell carcinoma(P<0.05,φ=0.446).The mean MVD value in ESCC was significantly higher than that in normal esophageal tissue(25.02±5.52 vs 12.09±4.55,P<0.05).MVD value in ESCC was closely correlated with lymph node metastasis.The mean MVD value in ESCC cases with positive CD40 and COX-2 expression was higher than that in those with negative CD40 and COX-2expression(26.37±6.02 vs 22.58±5.25.P<0.05).Western blot results showed that CD40 and COX-2 expression in Eca109 was higher than that in cultured normal esophageal epithelial cells (P<0.05).Conclusion:The expression of CD40 is involved in the carcinogenesis and progression of esophageal squamous cell carcinoma.CD40 may play an important role in the angiogenesis of ESCC through promoting COX-2 expression.