The role of lower esophageal sphincter (LES) in laryngopharyngeal reflux is controversial. In this study, we used an animal model to investigate the association between LES dysfunction and reflux laryngitis. Twelve healthy New Zealand albino rabbits (2.5-3.5 kg) were utilized in this study. The animals were divided into two groups. Eight rabbits underwent total cardiomyectomy to induce reflux, and the remaining four rabbits underwent a control sham operation. A laryngoscopy and a 24-hour intra-esophageal pH-metry were performed prior to surgery and again 2 and 8 weeks postsurgery. After the final laryngoscopy, all animals were sacrificed to obtain histological results. Total cardiomyectomy significantly increased the reflux index, the duration of the longest reflux episode and the total number of episodes that occurred in 24 h postsurgery. No significant difference was observed in the reflux finding score (RFS) between preoperative and 2-week postoperative rabbits (P = 0.11). But there was a statistically significant change in the RFS before and 8 weeks after the induction of reflux from 4.6 ± 0.9 to 8.3 ± 3.6 (P = 0.02). Submucous gland hyperplasia and inflammation were significantly increased in the reflux group compared to the control group. The results of this study suggest that chronic lower esophageal sphincter dysfunction is associated with reflux laryngitis in rabbits.

To optimize somatic cell nuclear transfer (SCNT) procedures in mini-pigs, the present study was designed to examine the effects of donor cell types and aphidicolin (APC) treatment on in vitro development of reconstructed embryos. Oviduct epithelial cells (OEC), ear fibroblast cells (EFC) and cumulus cells (CC) derived from mini-pigs were treated with serum starvation only or serum starvation followed by treatment of 0.1 µg/mL APC. The reconstructed embryos were cultured for 7 days to evaluate their developmental competency. Cleavage and blastocyst formation rates of reconstructed embryos derived from the OEC by APC treatment were significantly higher than the serum starvation (61.82% vs. 56.25%, 24.55% vs. 17.86%; P < 0.05). The cleavage rate from the EFC was significantly increased by APC treatment compared to serum starvation only (63.36% vs. 57.01%; P < 0.05). In the ooctyes with the CC, the reconstructed embryos could yield high blastocyst formation rate by APC treatment (29.63%; P < 0.05). In the presence of APC, CC gave rise to the highest cleavage and blastocyst formation rates among the three cell types. Therefore, our results suggest that treatment of CC with serum starvation plus APC prior to nuclear transfer is more suitable in SCNT of mini-pigs.

OBJECTIVE There has been a growing number of case reports on uncommon thyroglossal duct cyst (TDC) extending into the oropharynx and/or laryngopharynx, which has often been misdiagnosed. We therefore examined the unfamiliar radiological and clinical characteristics of the cases in question so that we could advance the current store of knowledge. METHODS From January 2005 to May 2009, all cases with TDC seen at a tertiary hospital, whose clinical data and images included CT and MRI, were included in this retrospective study. Cases with an uncommon thyroglossal duct cyst were chosen and divided to three different sub-types: intra-laryngeal, intra-lingual, and trans-laryngeal, according to the clinical syndrome and the sites of masses. RESULTS In this study, 250 cases with TDC were collected, 21 (8.40%) of which presented extending images. Five such cases were of the intra-laryngeal type, with a mass lying close to the foramen cecum or posterior of the hyoid bone; 2 were classified as intra-lingual type, with a cyst in the tongue; 14 were classified as trans-laryngeal type, with a tumor occurring below the hyoid bone. CONCLUSION Uncommon TDCs tend to extend into the range of the respiratory tract. This novel type of ingrowth could facilitate early correct diagnosis and the formulation of an appropriate treatment plan.

OBJECTIVE: Acid-sensing ion channels (ASICs) are members of the degenerin/epithelial sodium channel (DEG/ENaC) protein superfamily and play a critical role in acid-induced cell injury. In this study, we examined whether drugs such as amiloride that block ASICs could attenuate acid-induced apoptotic injury to articular chondrocytes. METHODS: Articular chondrocytes were isolated from Sprague-Dawley rats, and their phenotype was determined by toluidine blue and immunocytochemical staining. Articular chondrocyte viability assay was performed with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT). Apoptosis of chondrocytes was observed by the terminal deoxyribonucleotidyl transferase-mediated dUTP nick-end labeling method as well as propidium iodide labeling methods. Intracellular calcium ([Ca(2+)](i)) was analyzed by a Ca(2+)-imaging method. In addition, the expression levels of calpain and calcineurin in articular chondrocytes were examined by real-time PCR and immunocytochemical staining. The activity of caspase-3 was evaluated by spectrophotometric assays. RESULTS: Positive staining for glycosaminoglycan and collagen II was seen in articular chondrocytes. Blocking acid-sensing ion channels significantly decreased the cell death percentage and increased cell viability following acid exposure. After pretreated with amiloride, acid-induced [Ca(2+)](i) rises were reduced. Amiloride also inhibited calpain and calcineurin expression levels in acid-induced chondrocytes, and inhibited caspase-3 activity. CONCLUSION: The data presented in this study provided some experimental evidence that blocking ASICs could protect acid-induced apoptotic injury to chondrocytes.

α-amy gene amplified from barley genome was cloned into MCS of pGAP9K to generate pGAP9K-α-amy which was then transformed into Pichia pastoris GS115 by electroporation. Transformants with multi-copies and high expression for the foreign gene were selected on G418 containing plate and expression analysis. The fermentation was carried out in a 50 l bioreactor with 20 l working volume, using a high-density cell culture method by continuously feeding with 50% glycerol-0.8% PTM4 to the growing culture for 54 h at 30°C. Under the control of GAP promoter (pGAP), α-amy gene was constitutively expressed. At the end of the fermentation, the α-AMY expression reached 125 mg/l, while the biomass growth was 186 as measured by absorption of 600 nm. The secreted α-AMY was purified to 97.5% by SP-Sepharose FF ion-exchange chromatography and affinity purification. The recombinant α-AMY showed activity on hydrolysis of starch.

OBJECTIVE To explore the radiologic characteristics of lingual thyroglossal duct cyst relevant to diagnosis. In addition, to probe the feasibility of modified Sistrunk surgery for patients who experience recurrence after cystectomy or marsupialization under endoscopy. STUDY DESIGN Retrospective. METHODS Seven patients, three male and four female, who had been diagnosed with lingual thyroglossal duct cysts, had received cystectomy or marsupialization under endoscopy between one and eight times with cyst recurrence soon after every operation. All of these cases were evaluated by CT before proceeding with the latest modified Sistrunk surgery. During surgery, we routinely used methylene blue to trace the cysts, and then we used an enlarged Sistrunk procedure for such cases. RESULTS The CT images demonstrated that there were cysts located posteriorly to the tongue that had a close relationship with the hyoid bone and the foramen cecum. In addition, the results of the pathological examinations were consistent with thyroglossal duct cysts. During the postoperative follow up of 18 to 31 months, the patients did not show any symptoms or signs of recurrence. The results of their most recent examinations, two by laryngoscopy and five by CT scan, did not reveal any abnormality in their pars laryngea pharyngis. CONCLUSIONS CT could differentiate lingual thyroglossal duct cysts from the other cysts occurring at the lingual root. For those patients with lingual thyroglossal duct cysts that recur after cystectomy or marsupialization under endoscopy, the modified Sistrunk operation could be a suitable treatment option.

The present study reports the cloning and sequencing of lac2 from Bacillus subtilis. The gene is composed of 1542bp and encodes a 514-amino acid protein. The gene has 86% homology with a published laccase with GeneID 936023. The lac2 gene was deposited in GenBank as a new nucleotide sequence. This new sequence was cloned into the multiple cloning site of pPIC9K to generate pPIC9K-lac2, which was then transformed into Pichia pastoris GS115 via electroporation. The recombinant GS115 (pPIC9K-lac2) was grown initially in BMGY medium and transferred to BMMY to induce gene expression for 48h. The recombinant Lac2 protein shows laccase activity with α-naphthol and guaiacol as substrates. The optimal pH is between 3.2 and 4.7, and the optimal temperature is 25°C for enzyme reaction.

CONCLUSION The papers collected in our study demonstrated that there were no significant differences in the effectiveness of sound transmission and stability of the prosthesis between the titanium and non-titanium group. However, further randomized controlled trials with large sample sizes are needed to improve the estimation. OBJECTIVE To compare the effectiveness of sound transmission and stability of the prostheses in the titanium or non-titanium ossiculoplasty groups by meta-analysis for clinical implications and applications. METHODS Previous studies comparing the hearing status of patients with titanium and non-titanium prostheses were reviewed on PubMed, accompanied by a manual search of all the related references. The eligibility of the investigation was independently confirmed by two reviewers, who extracted all the data from the papers included accordingly for the standard meta-analysis. RESULTS The study included 12 eligible investigations covering 1388 patients, with the pooled risk ratio (RR) estimating the effectiveness of the postoperational prostheses between the titanium and non-titanium group, with a combined RR of 1.12 (95% confidence interval (CI): 0.98-1.27, p = 0.09), as well as their stability, with a combined RR of 1.47 (95% CI: 0.76-2.84, p = 0.26).

Parental effects are a major source of phenotypic plasticity. Moreover, there is evidence from studies with a wide range of species that the relevant parental signals are influenced by the quality of the parental environment. The link between the quality of the environment and the nature of the parental signal is consistent with the idea that parental effects, whether direct or indirect, might serve to influence the phenotype of the offspring in a manner that is consistent with the prevailing environmental demands. In this review we explore recent studies from the field of 'environmental epigenetics' that suggest that (1) DNA methylation states are far more variable than once thought and that, at least within specific regions of the genome, there is evidence for both demethylation and remethylation in post-mitotic cells and (2) that such remodeling of DNA methylation can occur in response to environmentally-driven, intracellular signaling pathways. Thus, studies of variation in mother-offspring interactions in rodents suggest that parental signals operate during pre- and/or post-natal life to influence the DNA methylation state at specific regions of the genome leading to sustained changes in gene expression and function. We suggest that DNA methylation is a candidate mechanism for parental effects on phenotypic variation.

Brain-derived neurotrophic factor (BDNF) and its receptor, tropomyosin-related kinase receptor B (TrkB), play a critical role in activity-dependent plasticity processes such as long-term potentiation, learning, and memory. It has been shown that BDNF exerts different or even opposite effects on behavior depending on the neural circuit. However, the detailed role of BDNF in memory process on the basis of its location has not been fully understood. Here, we aim to investigate the regional specific involvement of BDNF/TrkB in hippocampal-independent conditioned taste aversion (CTA) memory processes. We found region-specific changes in BDNF expression during CTA learning. CTA conditioning induced increased BDNF levels in the central nuclei of amygdala (CeA) and insular cortex, but not in the basolateral amygdala (BLA) and ventromedial prefrontal cortex. Interestingly, we found that the enhanced TrkB phosphorylation occurred at the time point before the increased BDNF expression, suggesting rapid induction of activity-dependent BDNF secretion by CTA learning. Moreover, targeted infusion of BDNF antibodies or BDNF antisense oligonucleotides revealed that activity-dependent BDNF secretion and synthesis in the CeA, but not the BLA, was respectively involved in the short- and long-term memory formation of CTA. Finally, we found that infusion of exogenous BDNF into the CeA could enhance CTA learning. These data suggest that region-specific BDNF release and synthesis temporally regulate different CTA memory phases through activation of TrkB receptors.