Decreased syntaxin17 expression contributes to the pathogenesis of acute pancreatitis in murine models by impairing autophagic degradation.

Acute pancreatitis (AP) is an inflammatory disease of the exocrine pancreas. Disruptions in organelle homeostasis, including macroautophagy/autophagy dysfunction and endoplasmic reticulum (ER) stress, have been implicated in human and rodent pancreatitis. Syntaxin 17 (STX17) belongs to the soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) subfamily. The Qa-SNARE STX17 is an autophagosomal SNARE protein that interacts with SNAP29 (Qbc-SNARE) and the lysosomal SNARE VAMP8 (R-SNARE) to drive autophagosome-lysosome fusion. In this study, we investigated the role of STX17 in the pathogenesis of AP in male mice or rats induced by repeated intraperitoneal injections of cerulein. We showed that cerulein hyperstimulation induced AP in mouse and rat models, which was characterized by increased serum amylase and lipase activities, pancreatic edema, necrotic cell death and the infiltration of inflammatory cells, as well as markedly decreased pancreatic STX17 expression. A similar reduction in STX17 levels was observed in primary and AR42J pancreatic acinar cells treated with CCK (100 nM) in vitro. By analyzing autophagic flux, we found that the decrease in STX17 blocked autophagosome-lysosome fusion and autophagic degradation, as well as the activation of ER stress. Pancreas-specific STX17 knockdown using adenovirus-shSTX17 further exacerbated pancreatic edema, inflammatory cell infiltration and necrotic cell death after cerulein injection. These data demonstrate a critical role of STX17 in maintaining pancreatic homeostasis and provide new evidence that autophagy serves as a protective mechanism against AP.

Effects of SPARCL1 on the proliferation and differentiation of sheep preadipocytes.

Important candidate genes that regulate lipid metabolism have the potential to increase the content of intramuscular fat (IMF) and improve meat quality. Secreted protein acidic and rich in cysteine like 1(SPARCL1) is a secreted glycoprotein with important physiological functions and is involved in the proliferation and differentiation of various cells. However, the role of the SPARCL1 gene in sheep preadipocytes and its regulatory mechanism is still unclear. In this study, we explored the effect of SPARCL1 on the proliferation and differentiation of sheep preadipocytes. The results showed that the expression level of the SPARCL1 gene is higher in fat tissue than in other tissues, and the gene was significantly increased on the 6th day of preadipocyte differentiation. In the preadipocyte proliferation stage, interference of SPARCL1 gene reduced cell viability and increased cell apoptosis. In preadipocyte differentiation stage, SPARCL1 overexpression significantly inhibited lipid droplets accumulation and triglyceride content by increasing Wnt10b, Fzd8, IL6, and ß-catenin and inhibiting PPARγ, C/EBPα, LPL, and IGF1 genes expression, whereas SPARCL1 deficiency significantly promoted cell differentiation by inhibiting ß-catenin and increasing GSK3ß, PPARγ, C/EBPα, and LPL. The results of this study suggest that SPARCL1 plays a negative role during preadipocyte differentiation and may become a novel target for regulating preadipocyte differentiation and improving IMF.Abbreviations:IMF: Intramuscular fat SPARCL1: Secreted protein acidic and rich in cysteine like 1 PPARγ: Peroxisome proliferator-activated receptor γ C/EBPα: CCAAT/enhancer-binding protein-α LPL: Lipoprotein lipase IGF1: Insulin-like growth factor 1 Wnt10b: Wnt family member 10B Fzd8: Frizzled class receptor 8 IL6: Interleukin 6 ß-catenin: Catenin beta interacting protein 1 GSK3ß: Glycogen synthase kinase 3 beta LRP5/6: Low-density lipoprotein receptor-related protein 5/6.

Protective effects of SOD2 overexpression in human umbilical cord mesenchymal stem cells on lung injury induced by acute paraquat poisoning in rats.

AIMS: To study the protective effects and mechanisms of human umbilical cord mesenchymal stem cells (hUCMSCs) and overexpression of antioxidant gene SOD2 on lung injury by establishing a rat model of paraquat (PQ)-induced lung injury. MAIN METHODS: The hUCMSCs cell line overexpressed SOD2 was established. After intraperitoneal injection of PQ solution (24 mg/kg) 3 h later, the different groups of hUCMSCs cell lines were injected through the tail veins of rats. Bronchoalveolar lavage fluid (BALF) was obtained to determine the protein level of inflammatory cytokines. Lung tissues were collected to test the wet/dry weight ratios (W/D), oxidative stress index and lung injury scores. Western blotting was used to detect SOD1, SOD2, HO-1, Nrf2, NF-κBp65 subunit, and cleaved caspase-3. KEY FINDINGS: After treatment with cells built on the basis of hUCMSCs, the protein levels of TNF-α, IL-8, and ICAM-1 in BALF decreased, and meanwhile in lung tissues, MDA content was reduced, GSH-Px activity was elevated, and lung W/D ratio decreased. Additionally, protein expression of NF-κB p65 subunit and activated caspase-3 in lung tissues was down-regulated, whereas expression of SOD1, SOD2, HO-1, and Nrf-2 were up-regulated. The results of HE staining showed that lung injury was significantly alleviated in the hUCMSC treated group. It is noticeable that hUCMSCs and SOD2-overexpressed hUCMSCs effectively reduced PQ-induced lung injury in rats, and moreover, hUCMSCs overexpressed SOD2 were more effective compared with hUCMSCs only. SIGNIFICANCE: Evaluation of the efficacy and analysis of mechanism in the treatment of PQ induced ALI by appliance of SOD2-overexpressed hUCMSCs will provide the proof from bench to bedside.

A comparison of three MDCT post-processing protocols: preoperative assessment of the ossicular chain in otitis media.

The aim of this study was to examine and assess comparative values of HRCT-based multiplanar reformation (MPR), volume rendering (VR) and virtual endoscope built on three-dimensional (3D) shaded-surface display (SSD-based CTVE) for detections of ossicular chain's damage in patients with otitis media. 70 human ears from 70 patients suffering by chronic otitis media or cholesteatoma, who were examined with a preoperative multi-slice computer tomography (MSCT) examination and tympanoplasty in our hospital were collected. The patients ossicular chains were reconstructed with the aforementioned three protocols and assessed via a three-point scoring system by three radiologists. Then, all the patients ossicular chains were reviewed by a surgeon and a radiologist via the same three-point scoring system used during surgeries at same time. By calculation, the Youden's index and coincidence rate were acquired without a significant difference for display of malleus. With regard to the incus, the Youden's index and coincidence rate of VR and MPR did not show any difference, however, both were higher than CTVE. For representation of the stapes, the accuracy of these three modalities is very low; especially, for the CTVE. In conclusion, both MPR and VR are relative robust, and CTVE is not effective for evaluation of small ossicular structures, particularly the stapes. Furthermore, the VR images are real 3D ones. Therefore, it could be the more valuable protocols for detection of the damage of ossicular chain in the patients with otitis media, and should be further applied in the future work.

Diversity and taxonomy of endophytic xylariaceous fungi from medicinal plants of Dendrobium (Orchidaceae).

Dendrobium spp. are traditional Chinese medicinal plants, and the main effective ingredients (polysaccharides and alkaloids) have pharmacologic effects on gastritis infection, cancer, and anti-aging. Previously, we confirmed endophytic xylariaceous fungi as the dominant fungi in several Dendrobium species of tropical regions from China. In the present study, the diversity, taxonomy, and distribution of culturable endophytic xylariaceous fungi associated with seven medicinal species of Dendrobium (Orchidaceae) were investigated. Among the 961 endophytes newly isolated, 217 xylariaceous fungi (morphotaxa) were identified using morphological and molecular methods. The phylogenetic tree constructed using nuclear ribosomal internal transcribed spacer (ITS), large subunit of ribosomal DNA (LSU), and beta-tubulin sequences divided these anamorphic xylariaceous isolates into at least 18 operational taxonomic units (OTUs). The diversity of the endophytic xylariaceous fungi in these seven Dendrobium species was estimated using Shannon and evenness indices, with the results indicating that the dominant Xylariaceae taxa in each Dendrobium species were greatly different, though common xylariaceous fungi were found in several Dendrobium species. These findings implied that different host plants in the same habitats exhibit a preference and selectivity for their fungal partners. Using culture-dependent approaches, these xylariaceous isolates may be important sources for the future screening of new natural products and drug discovery.

Sclerotial formation of Polyporus umbellatus by low temperature treatment under artificial conditions.

BACKGROUND: Polyporus umbellatus sclerotia have been used as a diuretic agent in China for over two thousand years. A shortage of the natural P. umbellatus has prompted researchers to induce sclerotial formation in the laboratory. METHODOLOGY/PRINCIPAL FINDING: P. umbellatus cultivation in a sawdust-based substrate was investigated to evaluate the effect of low temperature conditions on sclerotial formation. A phenol-sulfuric acid method was employed to determine the polysaccharide content of wild P. umbellatus sclerotia and mycelia and sclerotia grown in low-temperature treatments. In addition, reactive oxygen species (ROS) content, expressed as the fluorescence intensity of mycelia during sclerotial differentiation was determined. Analysis of ROS generation and sclerotial formation in mycelia after treatment with the antioxidants such as diphenyleneiodonium chloride (DPI), apocynin (Apo), or vitamin C were studied. Furthermore, macroscopic and microscopic characteristics of sclerotial differentiation were observed. Sclerotia were not induced by continuous cultivation at 25°C. The polysaccharide content of the artificial sclerotia is 78% of that of wild sclerotia. In the low-temperature treatment group, the fluorescent intensity of ROS was higher than that of the room temperature (25°C) group which did not induce sclerotial formation all through the cultivation. The antioxidants DPI and Apo reduced ROS levels and did not induce sclerotial formation. Although the concentration-dependent effects of vitamin C (5-15 mg mL(-1)) also reduced ROS generation and inhibited sclerotial formation, using a low concentration of vitamin C (1 mg mL(-1)) successfully induced sclerotial differentiation and increased ROS production. CONCLUSIONS/SIGNIFICANCE: Exposure to low temperatures induced P. umbellatus sclerotial morphogenesis during cultivation. Low temperature treatment enhanced ROS in mycelia, which may be important in triggering sclerotial differentiation in P. umbellatus. Moreover, the application of antioxidants impaired ROS generation and inhibited sclerotial formation. Our findings may help to provide new insights into the biological mechanisms underlying sclerotial morphogenesis in P. umbellatus.

Uncommon thyroglossal duct cyst with inward tendency: a recommendation regarding ingrowth type.

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.

Lingual thyroglossal duct cyst with recurrence after cystectomy or marsupialization under endoscopy: diagnosis and modified Sistrunk surgery.

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.

3D image of the middle ear ossicles: three protocols of post-processing based on multislice computed tomography.

The aim of this study was to examine and assess the comparative values of HRCT-based multiplanar reformation (MPR), volume rendering (VR) and virtual endoscope built on three-dimensional shaded-surface display (SSD-based CTVE) for evaluations of the ossicular chain. The normal pure tone audiograms, type-A tympanogram, and normal HRCT characteristics of 32 human ears of 18 patients were reviewed, whose ossicular chains were reconstructed with the three aforementioned protocols and assessed via the 3-point scoring system. The HRCT-based protocols could demonstrate a 3D image of the ossicular chain, except that of the footplate on the SSD-based CTVE. On the qualitative assessment, the efficacy of the MPR and VR, which were both superior to the SSD-based CTVE (P < 0.05), presented no statistical significance among the major and/or hyperdense structures (P > 0.05). As regards the lateral process of the malleus, VR was found to be significantly superior to the MPR and SSD-based CTVE (P < 0.05), both of which, however, showed no significant comparative differences (P > 0.05). Moreover, the three protocols in terms of efficacy were comparatively different in their representations of the anterior crus and footplates of the stapes, respectively (P < 0.05). On the MPR images, not all the images of the lenticular process were ideal; 20 of 32 cases were detected, but not defined. VR could be the more valuable protocol for the 3D reconstruction of the ossicular chain and ought to be more employed in future, especially for the education.

[CT multiplane reconstruction images of superior semicircular canal dehiscence syndrome].

OBJECTIVE: To evaluate the clinical application of multi-planar reformation (MPR) for the diagnosis of superior semicircular canal dehiscence syndrome. METHODS: A retrospective study was conducted on 9 patients who were diagnosed with SSCD syndrome in the Otology and Skull Base Surgery group of Fudan University. Three radiologists analyzed all the patients' 0.75 mm-collimated axial and coronal images and 0.75 mm-collimated MPR images, and they came up with the same results. RESULTS: There were 18 superior semicircular canal in the 9 patients, of whom 9 were intact and 9 were defective. All the defective superior semicircular displayed a definite dehiscence in all the MPR images, which indicated the sensitivity was 100%; however, 7 of the 9 defective superior semicircular canal were diagnosed as dehiscence in axial images, while 8 of the 9 were diagnosed in coronal images, but the sensitivities were 77.8% and 88.9% respectively. The results of the other 9 with intact superior semicircular canal displayed in the MPR, axial, and coronal images were also different. In the MPR images, they all displayed definite intact roof over the superior semicircular canal. There were 2 dehiscence in all axial and coronal images, and the specificities were 77.8%. CONCLUSION: The MPR image is more useful in diagnosis of superior semicircular canal dehiscence syndrome than that of the routine axial and coronal images.