[Evaluation of chemosensory function in patients with upper respiratory tract post-viral olfactory dysfunction].

Objective: To observe the changes of olfactory function, intranasal trigeminal nerve function and taste function in patients with upper respiratory tract post-viral olfactory dysfunction (PVOD), and to explore the correlation of chemosensory function. Methods: The clinical data of 42 patients with PVOD who visited to the Olfactory and Taste Center of Otorhinolaryngology Head and Neck Surgery Department of Beijing Anzhen Hospital from January to December of 2019 were analyzed retrospectively, including 20 males and 22 females, aging (48.86±11.47) years (x¯). Twenty subjects in normal control group were selected according to the sex ratio of PVOD patients. Sniffin' Sticks olfactory tests were performed on the subjects, including threshold test (T), discrimination test (D) and identification test (I), and the sum of the above three test scores was the TDI value. At the same time, olfactory event-related potentials (oERPs), trigeminal event-related potentials (tERPs) and taste function test were performed. According to the taste function test, the patients were divided into normal gustation (NG) group and gustatory dysfunction (GD) group. The results of olfaction, taste and intranasal trigeminal nerve function tests were compared among different groups, and the correlation analysis was carried out. SPSS statistical software was used for statistical analysis. Results: GD was present in 14 (33.3%) of 42 PVOD patients with a course of PVOD of 5 (3, 6) months (M (Q1, Q3)). The gustatory function of patients with PVOD was related to gender (r=0.565, P<0.001), smoking status (r=-0.512, P=0.001), duration (r=-0.357, P=0.020) and olfactory function (all P<0.05). The olfactory function of GD group was worse than that of NG group, and the differences of TDI value and T value between the two groups were statistically significant (10.25±4.58 vs 13.35±3.61, 1.54±0.66 vs 2.10±0.88, t value was 2.40 and 2.10 respectively, both P<0.05). The amplitudes of oERPs and tERPs were significantly lower in GD group than those in NG group (all P<0.05). Conclusion: In patients with PVOD, the subjective and objective olfactory function, intranasal trigeminal nerve function and taste function were decreased, and there was a correlation, suggesting that there was a synergistic effect between the chemosensory functions of PVOD patients.

[Analysis of upper airway structure development in children with different characteristics].

Objective: To analyze the development of upper airway in children with different characteristics. Methods: From June 2018 to June 2020, a total of 425 children younger than 16 years old who underwent head MRI examination and did not have sleep-disordered breathing were included in the study. The length of soft palate, tongue, upper airway, mental spine clivus, adenoid thickness and nasopharyngeal width were measured in the midsagittal plane of MRI image. Single factor variance analysis was used to compare the gender differences of upper airway parameters within certain age groups. Pearson correlation analysis was used to analyze the correlation between upper airway parameters and age. Results: The numbers of subjects in infant, young child, preschool, school age and adolescent group were 80, 86, 90, 90 and 79, respectively. There were 219 males, accounting for 51.5% of the study population. The adenoid thickness in the preschooler group was (1.26±0.26) cm, higher than that in the female group (1.15±0.20) cm (P=0.025). The upper airway length (5.89±0.60) cm and the ratio of upper airway length/mental spine-slope length (0.73±0.08) in males were higher than those in females [(5.31±0.45) cm and 0.67±0.07, respectively, P<0.05]. There was no gender difference in other upper airway parameters among different age groups (all P values>0.05). The length of upper airway, mental spine-slope, tongue, soft palate, the width of nasopharyngeal cavity and the thickness of adenoids were positively correlated with age (r=0.932, 0.912, 0.898, 0.705, 0.734 and 0.168, respectively), all P values<0.05. Adenoid thickness was positively correlated with age from birth to age 5 years (r=0.603, P<0.001), and negatively correlated with age after age 6 years (r=-0.259, P=0.001). Conclusion: There are gender differences in the development of upper airway structure in children of different ages.

[Study on the clinical characteristics of isolated congenital anosmia].

Objective: To retrospectively analysely the electrophysiological and imaging features of isolated congenital anosmia (ICA) and to assess the clinical phenotypic characteristics and classification of ICA. Methods: Clinical data of 30 ICA patients in Beijing Anzhen Hospital from 2012 to 2019 was retrospectively reviewed, including 13 males and 17 females, aged (35±19) years. The control group consisted of 30 healthy people from medical examination center, including 13 males and 17 females, aged (39±14) years. The clinical characteristics of ICA were analyzed using Sniffin' Sticks test, olfactory event-related potentials (oERPs), trigeminal event-related potentials (tERP) and olfactory pathway MRI. SPSS 17.0 software was used to compare the difference of olfactory function between the two groups. The correlation between olfactory bulb, olfactory sulcus structure and age was observed, and the clinical phenotype characteristics of ICA patients were analyzed. Results: The subjective olfactory function was completely lost in ICA patients. oERP was absent in all of the ICA patients, but showed normal N1 and P2 waves in controls. tERP could be evoked in 63.3% (19/30) of ICA patients, and signals in these patients showed higher amplitude in the N1 ((-10.33±6.93) µV vs (-5.11±2.71) µV, t=-10.113, P<0.01) and P2 ((+17.25±8.51) µV vs (+7.31±3.46) µV, t=5.443, P<0.01) waves than that of the controls. Olfactory bulbs were aplastic in 80.0% (24/30) of patients and hypoplastic in 20.0% (6/30) of patients. Fifty-six point seven percent (17/30) of patients had bilateral olfactory sulcus deletion while 43.3% (13/30) had dysplasia, and all of the patients exhibited a depth of olfactory sulcus less than 8 mm. Both the structure of olfactory bulbs and olfactory sulcus were not associated with age for ICA patients (r value was -0.174 and 0.325, respectively, all P>0.05). Conclusions: ICA patients show neurophysiologic deficits and some anatomic differences compared with healthy controls. The absence of oERP combining with a depth of olfactory sulcus less than 8 mm is the important indicator for clinical diagnosis of ICA. The structure of olfactory bulb may be a critical factor for clinical classification of ICA.

[Analysis of factors affecting olfactory disfunctions in patients with chronic rhinosinusitis with nasal polyps].

Objective: To analyze the factors affecting olfactory disfunctions in patients with chronic rhinosinusitis with nasal polyps (CRSwNP). Methods: This was a retrospective analysis. Eighty-eight patients with CRSwNP who underwent endoscopic sinus surgery in Beijing Anzhen Hospital from 2014 to 2018 were enrolled, including 22 males and 66 females, with the age of (48.1±11.3) years old(Mean±SD). Sniffin' Sticks olfactory test, Lund-Mackay score and modified sinus CT olfactory cleft score, nasal resistance and acoustic reflex examination, blood routine and blood biochemistry test, serum specific IgE test were performed before surgery and nasal polyps of all patients were collected for eosinophil count during surgery. According to bilateral total TDI score, the patients were divided into normal olfactory function group and olfactory disfunction group. The clinical baseline data were compared between the two groups. According to the results of single factor analysis, factors which were significant different between the two groups and clinically useful indicators were further included in the multivariate Logistic regression model analysis, then a model predicting olfactory disfunction in patients with CRSwNP was initially established. P<0.05 was considered statistically significant. Results: Among 88 patients with CRSwNP, 32 (36.4%) patients were with normal olfaction and 56 (63.6%) patients were with olfactory disfunction, including 40 (45.5%) of hyposmia and 16 (18.2%) of anosmia. Tissue eosinophil count, blood eosinophil percentage and blood urea concentration had significant difference between the two groups (12.7[2.0, 52.3]/HP (M[P(25), P(75)]) vs 38.6[16.2, 87.0]/HP, 2.75[1.60, 4.80]% vs 4.35[2.50, 6.60]%, (5.56±1.15) mmol/L vs (4.98±1.33) mmol/L, all P<0.05). Modified sinus CT olfactory cleft score and Lund-Mackay score except for ostiomeatal complex score were statistically significant between the two groups (all P<0.05). Multivariate Logistic regression analysis showed that the bilateral and total olfactory cleft score and blood urea concentration were statistically significant, in addition, the bilateral and total olfactory cleft score was a risk factor (OR=2.108, 95%CI: 1.407-3.159, P<0.001) and blood urea within a certain concentration was a protective factor (OR=0.461, 95%CI: 0.240-0.884, P=0.020). Further studies found that the area under the ROC curve of the model with tissue eosinophil count, blood eosinophil percentage, bilateral and total olfactory cleft score, total inspiratory volume and blood urea concentration was 0.888 (P<0.01), which had good predictive value for olfactory disorders in CRSwNP. Conclusions: The modified sinus CT olfactory cleft score is closely related to the olfactory disorders in patients with CRSwNP. A certain degree of elevated blood urea concentration may have a protective effect on the olfactory function of patients with CRSwNP.

[Effect of endoscopic sinus surgery on olfactory function in patients with chronic rhinosinusitis with nasal polyps].

Objective:To evaluate the effect of surgery on olfactory function in patients with chronic rhinosinusitis with nasal polyps（CRSwNP） by subjective and objective olfactory tests. Method:This was a retrospective study. Forty patients with CRSwNP who underwent endoscopic sinus surgery（ESS） from 2015 to 2017 in Beijing Anzhen Hospital were enrolled. Postoperative time was 6-46 months. The patients were followed up in October 2018 and examined using Sniffin' sticks olfactory test and olfactory event-related potentials （oERP）. The polyps collected during surgery were performed to eosinophil count and percentage calculation. They were divided into eosinophilic CRSwNP（ECRS） and non-eosinophilic CRSwNP（NECRS）. The subjective and objective olfactory functions between the two groups were compared before and after surgery respectively and the paired T test was performed between the postoperative and the preoperative Sniffin' sticks olfactory test. Covariance analysis was used to adjust the effect of different postoperative time on postoperative olfactory recovery. Result:There were 21（52.5%） ECRS and 19（47.5%） NECRS patients of the 40 patients with nasal polyps. There was statistical difference in the posterior ethmoid score and the posterior olfactory cleft score of CT. According to the criterion of total TDI increased more than 5.5, olfactory function in 21（52.5%） patients had improved. In addition, there was a significant improvement in olfactory function in ECRS group either in unilateral T/TDI or bilateral T/D/TDI, but only unilateral T/TDI increased in NECRS group. There was no significant difference in Sniffin' sticks olfactory test between the two groups, but there was a statistically significant difference in the latency of oERP after surgery. Conclusion:ESS could improve olfactory function in patients with CRSwNP by more than 50%. Bilateral olfactory improvement in ECRS was better than that in NECRS, but olfactory function in postoperative ECRS was still lower than that in NECRS. oERP can more objectively and accurately reflect the severity of olfactory disorders associated with eosinophilic inflammation.

[Correlation of tissue eosinophil count and modified CT olfactory cleft score and olfactory function in patients with nasal polyps].

Objective:To investigate the effect of tissue eosinophil count on olfactory function in patients with chronic sinusitis with nasal polyps（CRSwNP）. Method:We prospectively selected 59 patients with CRSwNP. All the patients were not treated with oral or topical glucocorticoids for at least 1 month at the time of enrollment. All the nasal polyps were taken under local anesthesia in the outpatient department for eosinophil count and percentage calculation. The patients were divided into eosinophilic CRSwNP（ECRS group） and non-eosinophilic CRSwNP（non-ECRS group）. Baseline data was compared between the two groups. Spearman correlation analysis was performed on tissue eosinophil count and Lund-Mackay score, modified olfactory cleft scores, Sniffin' Sticks olfactory test, olfactory evoked potential peak latency and amplitude respectively. Result:Thirty-five of 59 patients with CRSwNP completed olfactory evoked potential test, from whom we collected stable waveforms. The ECRS group had a higher olfactory cleft area CT score, ethmoid sinus CT score and blood eosinophil count and percentage; there was a significant difference between the ECRS group and the non-ECRS group in single and bilateral odor test threshold（T value）（P=0.017）. There was no significant correlation between tissue eosinophil count and subjective olfactory VAS score（P>0.05）. Tissue eosinophil count was related to the bilateral T value in Sniffin' Sticks olfactory test（r=-0.322, P=0.013）, anterior and posterior olfactory cleft area scores（r=0.431 and 0.415, respectively, P=0.001） and olfactory evoked potential N1 latency（r=0.504, P=0.001） and P2 latency（r=0.374, P=0.020）, but not related to Lund-Mackay scores. In addition, there was a significant correlation between the unilateral T score in Sniffin' Sticks test and the unilateral olfactory evoked potential N1 latency（r=-0.505, P=0.002）. Conclusion:The increase of tissue eosinophilia was closely related to olfactory disfunctions in CRSwNP. It could be reflected by modified CT olfactory cleft score, Sniffin' Sticks olfactory test T value and oERP peak latency. In addition, T value was negatively consistent with latency of oERP N1 peak.

[Efficacy and associated factors of olfactory training in the treatment of olfactory dysfunction].

Objective: To explore the clinical effects and the influence factors of olfactory training in the treatment of olfactory dysfunction. Methods: A total of 86 patients with olfactory dysfunction (49 post-infectious and 37 post-traumatic) in Beijing Anzhen Hospital during Dec 2016 to May 2017 were recruited in this prospective study. The clinical data of patients were analyzed, including gender, age, body mass index (BMI), course of disease, smoking history, drinking history, diabetes history, hypertension history, hyperlipidemia history, and anxiety visual analogue score (VAS). All patients were treated with olfactory training for 16 weeks, and all of them underwent Sniffin' Sticks olfactory test before and after treatment, which was evaluated by composite threshold-discrimination-identification score (TDI). SPSS 23.0 software, paired t test and univariate and multivariate Logistic regression analysis were used to analyze the data. Results: Eighty patients received treatment, including 46 post-infectious olfactory dysfunction and 34 post-traumatic olfactory dysfunction. After olfactory training, the total scores of TDI increased with statistically significant (18.3±8.6 vs 13.6±7.4, t=-6.158, P<0.05). The overall efficacy was 40% (32/80). The effective rate were 45.7% (21/46) in post-infectious olfactory dysfunction and 32.4% (11/34) in post-traumatic olfactory dysfunction respectively, with no statistically significant difference (χ(2)=1.441, P=0.230). Logistic regression analysis showed that the course of disease was an influence factor in the clinical curative effect (OR=0.881, 95%CI: 0.799-0.973, P=0.012). In patients with less than a year of olfactory dysfunction, the olfactory function improved obviously with the efficiency of 50.9% (29/57). Conclusion: Sixteen weeks of olfactory training provides a significant therapeutic effect on the post-infectious and post-traumatic olfactory dysfunction, and the olfactory training can achieve better therapeutic effects at the early stage.

[The preliminary study of white matter integrity in patients with olfactory dysfunction].

Objective: To investigate the white matter integrity in patients with olfactory dysfunction using diffusion tensor imaging (DTI). Methods: Twenty-one patients with olfactory dysfunction and sixteen age, sex and level of education matched normal subjects were recruited in this study. Sniffin' Sticks olfactory test was performed to evaluate the olfactory function of all subjects. We acquired diffusion tensor images with a echo planar imaging (EPI) sequence from all subjects on a 3T scanner. The fractional anisotropy (FA) images were performed using DTI-studio, and bilateral piriform cortex, orbitofrontal cortex, hippocampus and insula cortex adjacent white matter as well as capsula interna were delineated from the FA images as the region of interest associated with olfactory (ROI(awo)) manually. Independent sample t test analysis was used to compare the FA value of all ROI(awo) between the controls and patients. Results: In olfactory dysfunction group, the FA value of adjacent white matter of right piriform cortex and orbitofrontal cortex were significantly lower than those of control group (0.42±0.05 (x±s) vs 0.45±0.05, 0.43±0.06 vs 0.49±0.07, t value was 2.32, 2.79, respectively, all P<0.05). The FA value of adjacent white matter of left piriform cortex and orbitofrontal cortex had no significant difference compared with those of control group (0.43±0.05 vs 0.45±0.04, 0.44±0.04 vs 0.47±0.06, t value was 1.65, 1.37, respectively, all P>0.05). The FA value of the adjacent white matter of bilateral hippocampus, insula cortex and capsula interna had no significant difference compared with those of control group, neither (0.45±0.08 vs 0.44±0.08, 0.45±0.09 vs 0.44±0.10, 0.41±0.08 vs 0.39±0.07, 0.41±0.07 vs 0.38±0.05, 0.64±0.08 vs 0.63±0.08, 0.64±0.07 vs 0.63±0.07, t value was 0.30, 0.15, 0.88, 1.34, 0.14, 0.35, respectively, all P>0.05). Conclusions: The patients with olfactory dysfunction showed abnormal white matter connection in the major primary and secondary olfactory cortex. The reduced white matter integrity in ROI(awo) might contribute to the pathogenesis of olfactory dysfunction.

[MicroPET-CT study of the association between odor stimulation and olfactory related brain activation in rats].

Objective: Using (18)F-fluorodeoxyglucose ((18)F-FDG) and microPET-CT to test the feasibility of (18)F-FDG PET-CT for validation of olfactory function of rats with standard phenethyl alcohol (PEA) and isovaleric acid (IVA) odors stimulation. To verify the possibility of (18)F-FDG PET-CT as a new objective examination method for olfactory function. Methods: Six healthy Sprague-Dawley (SD) male rats were selected with a weight of 250-300 g. First of all, buried food pellet test (BFT) was used to confirm the normal olfactory function of rats. Then in the next 3 days, after the intravenous injection of (18)F-FDG (18 MBq/100 g), awaken rats were placed in a ventilated plexiglas cage for 30 min. Subsequently, pure air (the first day), PEA (the second day) and IVA (the third day) were delivered. After odor stimulation for 30 min, rats were performed by a static PET-CT under anesthesia. Images reconstructed were assessed by SPM method and analyzed by VBM method. Data was analysied by paired t test. Results: Activation regions of rat's brain after PEA stimulation included bed nucleus and insula. Activation regions of rat's brain after IVA stimulation included olfactory bulb, anterior olfactory nucleus, amygdala, entorhinal cortex, olfactory cortex, piriform cortex, insula, prefrontal cortex, cingulate cortex and bed nucleus (P<0.005, Ke>20 voxels). Conclusions: Through microPET-CT, we can observe that olfactory stimulation with different odors can induce metabolic activation in different regions of rat's brain, which was in concordance with olfactory regions. The olfactory related brain regions of rats have strong responses to odor stimulation of IVA.

[Assessment of the brain volume alterations in patients with hyposmia based on voxel-based morphometry].

Objective: To investigate the brain volume alterations in patients with hyposmia using voxel-based morphometry (VBM) and to correlate these alterations with the degree and duration of hyposmia. Methods: Forty patients with hyposmia from Department of Otorhinolaryngology Head and Neck Surgery, Beijing Anzhen Hospital since 2013 to 2016 and forty age and sex matched normal subjects were recruited in this study. Sniffin' Sticks olfactory test was performed to evaluate the olfactory function of all subjects. We acquired T1-weighted magnetic resonance images from all subject on a 3T scanner. VBM was performed using VBM8 toolbox and SPM8 in a Matlab environment. Independent sample t test analysis was used to compare the volume of gray and white matter between the controls and patients. In addition, the Pearson or Spearman correlation analysis was used for the correlation between the voxel value of cerebral volume alterations area and the degree and duration of hyposmia on patients (threshold discrimination identification, TDI). Results: Compared with the controls, patients showed significantly decreased volume in the gray matter of right orbitofrontal cortex (number of voxel in clusters was 226, t=-4.46, P<0.001, uncorrected). There was negative correlation between decreased gray matter volume of the right orbitofrontal cortex with significantly decreased area and the TDI results (r=-0.40, P=0.01), but positive correlation with duration of hyposmia (r=0.37, P=0.02). Conclusions: The patients with hyposmia show gray matter atrophy of the right orbitofrontal cortex. The duration may be an important risk factor for decreased gray matter in patients with hyposmia.

[Intervention study of compassion fatigue of oncology nurses in Balint group activities].

Objective: To explore the effectiveness of Balint group on compassion fatigue among oncology nurses. Methods: From January to December 2016, 35 oncology nurses from one general hospital were enrolled. 18 cases were allocated in the observation group and 17 cases in the control group by computer randomization. Nurses in the observation group were received a total of 8 times Balint Group activities, with 2 times a month and 1.5 hours each time, which is aiming to discuss difficult cases encountered in the clinic to help nurses have a deeper experience and a better understanding of the emotions and behaviors, fantasies and needs between nurse-patient interaction. Nurses in the control group without intervention. All the Participants were requested to complete the survey of the Professional Quality of Life (ProQOL) , the Jefferson Scale of Empathy-Health Professionals (JSE-HP) , and the General Health Questionnaire (GHQ) at pre and post intervention. Results: Before intervention, two group of nurses in age, working years, marriage, and education were not statistically significant (P>0.05). There was no difference in the scores of empathy, compassion satisfaction, secondary traumatic stress, and General health (P>0.05). After intervention, the scores of JSE-HP and its three dimensions of perspective taking, emotional care, and trans-positional consideration in the observation group had significantly higher than the control group (P<0.05). The level of compassion satisfaction in the observation group was higher, and the general health status was lower (P<0.05). There was no statistically difference in the level of burnout and secondary traumatic stress between two groups (P>0.05) . Conclusion: Balint group has a positive role in promoting nurses'empathetic skills, compassion satisfaction and mental health.

[Clinical treatment effect of glucocorticoids and extract of ginkgo biloba on post-viral olfactory dysfunction].

Objective:To observe the effect of ginkgo biloba extraction combined with glucocorticoids on postviral olfactory dysfunction.Method:Forty-two patients were diagnosed as postviral olfactory dysfunction. All patients underwent olfactory test, including T&T test and Sniffin Sticks test before and after treatment. The treatment lasted up to 3 months based on effectiveness. The results of olfactory test were recorded every month.Result:Twenty patients received the treatment with prednisone acetate. T&T test showed that the effective and improvement rate of the treatment with prednisone acetate were 25.00%(5/20) and 45.00%(9/20),respectively; Sniffin Sticks test showed that the effective and improvement rate of the treatment were 20.00%(4/20)and 50.00%(10/20),respectively. Twenty-two patients received the treatment combined with extract of ginkgo biloba. T&T test showed that the effective and improvement rate of the treatment with prednisone acetate were 31.82%(7/22)and 50.00%(11/22),respectively; through Sniffin Sticks test showed that the effective and improvement rate of the treatment were 27.27%(6/22)and 54.55%(12/22),respectively.Conclusion:Olfactory function in patients with postviral olfactory dysfunction was improved with two therapy. There was no significant difference on effect between the two therapeutic groups, but the effect of combination of extract of ginkgo biloba was better than the effect of prednisone acetate. Prolong duration of treatment is help for the recovery of the olfaction.

[Differences in clinical features of post-traumatic olfactory dysfunction and non-post-traumatic olfactory dysfunction: a follow-up study].

Objective:To analyze the clinical features and recovery rate of post-traumatic olfactory dysfunction (PTOD) in Chinese adults in a case control follow-up study. Method:The clinical data of 202 patients who were diagnosed with olfactory dysfunction between January 2015 and December 2016 and followed up for 14 to 473 days were analyzed in this retrospective study. The patients were divided into those with PTODs (PTOD group) and those without PTODs (non-PTOD group). The two groups were compared with regard to age (years), sex, olfactory function (Sniffin' sticks), gustatory function (triple drop method), chemosensory evoked potentials, and magnetic resonance imaging (MRI) characteristics of olfactory pathways. The recovery rate of PTOD was evaluated by Sniffin' sticks and triple drop method. Result:Patients in the PTOD group (40±11 years) were significantly younger than those in the non-PTOD group (47±15 years), whereas the number of men and women was similar in both groups. The mean TDI score (Sniffin' sticks) was significantly different between the PTOD (12±5) and non-PTOD (19±8) groups (P<0.05). The mean oERP P2 latency was significantly shorter for the non-PTOD group (418±64 ms) than for the PTOD group (483±82 ms, P<0.05). There were no significant differences in the mean oERP N1 latency, N1 amplitude, P2 amplitude, mean tERP P2 latency and MRI between the two groups. After the follow-up period, 8.9% (5/56) and 5.4% (3/56) patients in the PTOD group exhibited an improvement in olfactory function and gustatory function, respectively. Conclusion:PTOD should be considered a type of disability that can lead to serious accidents, and an adequate understanding of its clinical features and etiologies is critical for appropriate diagnosis and treatment and for improving the prognosis of treatment. The rate of recovery of olfactory function is higher than that of gustatory function in patients with PTOD; further investigations are required in this regard.

N-stearoyltyrosine protects primary neurons from Aß-induced apoptosis through modulating mitogen-activated protein kinase activity.

N-stearoyltyrosine (NsTyr), an anandamide (AEA) analogue is similar to AEA not only structurally but also in terms of biological activity. Since A beta-induced neuronal injury triggers the activation of mitogen-activated protein kinase (MAPK) pathways and the induction or activation of pro- and anti-apoptotic proteins, in the present study we aimed to assess the protective effect of NsTyr against A beta induced neuronal apoptosis. Cell viability and neuronal injury were respectively measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay and lactate dehydrogenase (LDH) assay. Hoechst staining and flow cytometric assessment were used to evaluate cell apoptosis. The anti-apoptotic mechanism involved MAPK phosphorylation and Bcl-2/Bax expression was investigated. The best neuroprotective effect on A beta 25-35-induced neuronal apoptosis was observed in the presence of NsTyr (1 microM). NsTyr exerted anti-apoptotic effect at least partly via activating p-ERK-Bcl-2 but suppressing p-p38-Bax pathways. Moreover a dynamic balance between p-ERK and p-p38 MAPK pathways in NsTyr-induced neuronal protection suggested an interaction between them. Our results indicated the neuroprotective effect of NsTyr on A beta 25-35-induced neuronal injury was at least partly due to anti-apoptosis and raised the possibility that NsTyr might reduce neurodegenerative disorders.

Developmental and glucocorticoid regulation of surfactant protein mRNAs in preterm lambs.

Glucocorticoid treatment increases content of surfactant protein (SP) A and SP-B in lung tissue and lavage fluid of preterm lambs. To investigate this process, we determined the ontogeny and glucocorticoid induction of SP mRNAs. In separate treatment protocols, each with its own controls, sheep were injected with betamethasone 15 h, 48 h, or weekly for 1-4 doses before preterm delivery. Using ovine SP cDNAs, we found an increase equal to or more than threefold in basal levels of all three SP mRNAs between 125 days and term. After betamethasone treatment, SP-B and SP-C mRNA levels increased by 15 h and all SP mRNAs were elevated after 24 h (>/=2-fold); mRNA levels in fetuses delivered 1-3 wk after betamethasone were not different from control. We conclude that in vivo betamethasone rapidly induces a coordinated increase in SP mRNAs, which is fully reversible within 7 days despite repetitive doses of betamethasone. Similar increases in mRNA and protein contents for SP-A and SP-B suggest that glucocorticoid regulation of these SPs in vivo is primarily pretranslational.

A unique new model to study the effects of urinary diversion in the developing rabbit bladder.

PURPOSE: Little is known about the developmental effects of high urinary diversion and bladder defunctionalization in infancy. Although clinical experience shows that a poorly functional bladder may result from urinary diversion in infancy, the mechanisms of change and specific bladder wall alterations have not been well characterized. We hypothesized that cyclic filling and emptying are necessary for normal bladder development. To investigate this important question we created a new animal model. MATERIALS AND METHODS: We designed a new method of hemibladder urinary diversion in 3-week-old New Zealand white rabbits. After vertical midline bladder division half of the bladder was formed into a functional reservoir, which remained in continuity with the ipsilateral ureter and urethra. The other bladder half was defunctionalized and isolated from the urine flow by ureteral ligation. Diversion was created for 3, 7, 14 and 28 days. Urodynamic evaluation was done in the functionalized hemibladders and age matched normal rabbit bladders to test the validity of the functionalized hemibladder as an internal control. Functional and defunctionalized hemibladders as well as age matched, nonoperated normal rabbit bladders were weighed, sectioned and stained to demonstrate muscle and connective tissue components. RESULTS: In 22 of the 27 healthy rabbits (81%) good quality diverted and functional bladder specimens were obtained after diversion. Defunctionalized hemibladders grew more slowly than functionalized bladders and normal age matched control bladders. Histological staining of the bladder wall demonstrated increased connective tissue between the muscle bundles within the diverted specimens than in functional bladders. CONCLUSIONS: Our successful model of urinary diversion may be used to study the developmental and histological effects of urinary diversion in the young bladder. Bladder growth and histological appearance are altered when the stimulus of cyclic filling and emptying is removed. Further studies using this model are warranted to define fully bladder changes that result from diversion and investigate the mechanism of the observed changes.

Effect of cryopreservation and histocompatibility on type I procollagen gene expression in aortic valve grafts.

BACKGROUND: Allograft valves are excellent substitutes for diseased or absent valves but undergo primary tissue degeneration. Fibroblast viability may determine resistance to valve deterioration. This study evaluated gene expression for procollagen by valve grafts and studied the effects of cryopreservation and histocompatibility on this property. METHODS AND RESULTS: Fresh and cryopreserved rat aortic valves were implanted heterotopically into syngeneic or allogeneic recipients. Nonviable, cryothermally injured valves were used as negative controls. The grafts and native aortic roots were excised 3 days after implantation. Northern hybridization with a human procollagen alpha 1 (I) complementary DNA probe was used to assess the expression of type I procollagen mRNA. The content of procollagen mRNA relative to 18S ribosomal RNA was evaluated by means of scanning densitometry. In situ hybridization was used to locate the areas of procollagen mRNA expression in the grafts. Both fresh and cryopreserved grafts exhibited greater expression than the native valve. This increase in expression was observed in both syngeneic and allogeneic grafts, but not in the negative control group. In situ hybridization showed a strong signal for procollagen in the aortic wall and a weak signal in the leaflet and myocardium in the viable grafts and in native tissues. CONCLUSIONS: Regardless of preservation or allogenicity, fibroblast viability in aortic valve grafts persists after implantation. Increased gene expression for procollagen suggests a capacity for repair and regeneration.

Developmental regulation of collagen differential expression in the rabbit bladder.

PURPOSE: It has been suggested that the amount and relative ratios of different types of collagen influence bladder compliance. To understand the mechanisms regulating collagen synthesis and degradation in the bladder we investigated the gene expression of collagen types I and III in the rabbit bladder during normal development. MATERIALS AND METHODS: New Zealand white rabbits ages fetus to adult were used for this study. The mid portion of the bladder wall was harvested. Northern blot hybridization was performed to analyze quantitatively collagen types I and III messenger (m) ribonucleic acid (RNA), and in situ hybridization was done to localize the messages. Corresponding protein distributions were obtained by immunohistochemical staining. RESULTS: Types I and III collagen are developmentally regulated at the level of gene expression. The temporal and spatial distribution of the mRNA transcripts of alpha 1(I) and alpha 1(III) correlates with extracellular protein deposition. Differential distribution of the mRNA transcripts is also developmentally regulated. CONCLUSIONS: This study characterizes the relationship between collagen gene expression and normal rabbit bladder development. The expression of alpha 1(I) and alpha 1(III) alters as the bladder grows. High levels and nonconcordant up and down regulation of different types of collagen mRNA during early development demonstrate the complexity of the extracellular matrix in a young bladder. This observation may be important in our understanding of the injury response in a developing versus mature bladder.

Collagen types I and III localization by in situ hybridization and immunohistochemistry in the partially obstructed young rabbit bladder.

PURPOSE: The objective of the study was to investigate the alteration of collagen expression patterns at the transcription and translation levels during partial outlet obstruction and to compare changes in collagen types I and III gene expression, and protein deposition during early and subacute phases of obstruction. MATERIALS AND METHODS: Following 1, 2, 7, 14 and 21 days of partial bladder obstruction in 2-week-old rabbits bladder tissues were evaluated for collagen types I and III messenger ribonucleic acid localization by in situ hybridization and for protein localization by immunohistochemical study. RESULTS: There is up regulation of collagen types I and III gene expression after obstruction. Up regulation of collagen gene expression immediately after obstruction is within the lamina propria and the message moves into the muscular layer as obstruction progresses. Protein deposition for both collagen types is analogous to that of gene expression. CONCLUSIONS: Up regulation of collagen types I and III gene expression, and increased protein deposition after partial obstruction demonstrate that the alteration in collagen synthesis is at least partially transcription regulated. This alteration starts from superficial layers of the bladder wall and becomes deeper as the pathological process progresses.