OBJECTIVE To discover a small- molecule activator of ULK1 for Parkinson disease treatment and exploreits potential mechanisms. METHODS Candidate ULK1 activator was found by using structure-based design and high-through put screening, then modified by chemical synthesis and screened by kinase and autophgic activities.The amino acid residues that key to the activation site of the best candidate ULK1 activator (BL-918) were determined by site-directed mutagenesis, as well as in vitro kinase assay, ADP- Glo kinase assay and surface plasmon resonance (SPR) analysis. The mechanisms of BL- 918 induced cytoprotective autophagy were investigated by electron microscopy, fluorescence microscopy, Western blotting, co-immunoprecipitation assay, siRNA and GFP-LC3 plasmid transfections. The therapeutic effect of BL- 918 was determined by MPTP- mouse model, including behavioral tests, the levels of dopamine and its derivatives, as well as immunofluorescence and Western blotting. The toxicity of BL-918 was assessed by blood sample analysis and hematoxylin-eosin staining. RESULTS We discovered a small molecule (BL-918) as a potent activator of ULK1 by structure-based drug design. Subsequently, some key amino acid residues (Arg18, Lys50, Asn86 and Tyr89) were found to be crucial to the binding pocket between ULK1 and BL- 918, by site- directed mutagenesis. Moreover, we found that BL- 918 could induce autophagy via the ULK complex in neuroblastoma SH-SY5Y cells. Intriguingly, this activator displayed a cytoprotective effect on MPP +-treated SH-SY5Y cells, as well as protected against MPTP-induced motor dysfunction and loss of dopaminergic neurons by targeting ULK1- modulated autophagy in mouse models of PD. CONCLUSION We discovered a novel ULK1 activator (BL-918) that potently activated ULK1. This activator could induce cytoprotective autophagy via the ULK1 complex in SH- SY5Y cells, and also exerted its neuroprotective effects by targeting ULK1- modulated autophagy in a MPTP- induced PD mouse model, which may serve as a candidate drug for future PD therapy.

Armillariella tabescens EJLY2098 was induced to produce ?-mannanase with konjac fine flour (Amorphopallus rivieri) as single carbon source. This induced enzyme was then purified using DEAE ion exchange chromatography and named atMAN47. Zymologic analysis showed that the molecular weight of this ?-mannanase was approximately 47 kD. The enzyme was stable when pH ranged from 5.0 to 6.5 and could be activated by Na+ and Ba2+. With an optimal temperature of 50?C. Action mode analysis of TLC revealed that the enzyme belonged to the endo-?-mannanase family. Being a meta-acid endo-?-mannanase, it was suitable to be applied to feed industry with a promising future as an enzyme preparation.

OBJECTIVETo compare the surface roughness of nanofilled dental composite resin and microhybrid composite resins after curing and polishing.

METHODSA nanofilled composite (Z350) and 4 microhybrid composites (P60, Z250, Spectrum, and AP-X) were fabricated from the lateral to the medial layers to prepare 8 mm×8 mm×5 mm cubical specimens. The 4 lateral surfaces of each specimens were polished with abrasive disks (Super-Snap). Profilometer was used to test the mean surface roughness (Ra) after polishing.

RESULTSP60 had the lowest Ra (0.125∓0.030 µm) followed by Z250 and Spectrum. The Ra of Z350 (0.205∓0.052 µm) was greater than that of the other 3 resins, and AP-X had the roughest surfaces. Under scanning electron microscope, the polished faces of P60 resin were characterized by minor, evenly distributed particles with fewer scratches; the polished faces of Z350 presented with scratches where defects of the filling material could be seen.

CONCLUSIONThe nanofilled composite Z350 has smooth surface after polishing by abrasive disks, but its smoothness remains inferior to that of other micro-hybrid composite resins.

Acrylic Resins ; Composite Resins ; Dental Materials ; Dental Polishing ; Materials Testing ; Polyurethanes ; Surface Properties

OBJECTIVETo evaluate the effect of thermal cycling on surface microstructure of different light-curing composite resins.

METHODSA nanofilled composite (Z350) and 4 microhybrid composites (P60, Z250, Spectrum, and AP-X) were fabricated from lateral to center to form cubic specimens. The lateral surfaces were abrased and polished before water storage and 40 000 thermal cycles (5/55 degrees celsius;). The mean surface roughness (Ra) were measured and compared before and after thermal cycling, and the changes of microstructure were observed under scanning electron microscope (SEM).

RESULTSSignificant decreases of Ra were observed in the composites, especially in Spectrum (from 0.164±0.024 µm to 0.140±0.017 µm, P<0.001) and Z250 (from 0.169±0.035 µm to 0.144±0.033 µm, P<0.001), whose Ra approximated that of P60 (0.121±0.028 µm) with smoothly polished surface. SEM revealed scratches and shallower pits on the surface of all the 5 resins, and fissures occurred on Z350 following the thermal cycling.

CONCLUSIONWater storage and thermal cycling may produce polishing effect on composite resins and cause fissures on nanofilled composite resins.

Composite Resins ; Dental Polishing ; Light-Curing of Dental Adhesives ; Materials Testing ; Surface Properties ; Temperature

BACKGROUNDRhabdomyosarcoma (RMS) is an uncommon malignancy of the breast. The aim of this study was to summarize its clinicopathologic features and biological behavior.

METHODSFive primary or secondary breast RMSs were collected. Their clinicopathological characteristics and all published literature about breast RMS were reviewed. Immunohistochemical study of desmin, myogenic differentiation 1 (MyoD1), myogenin, leukocyte common antigen (LCA), vimentin, cytokeratin (AE1/AE3), E-cadherin, neuron specific enolase (NSE), CD99, chorioallantoic membrane 5.2 (CAM5.2) and epithelial membrane antigen (EMA) expression were performed.

RESULTSThe five patients were all female with ages ranging from 16 to 46 years old (mean, 30 years). Three were metastatic breast RMSs, two embryonal and one solid variant alveolar, with the primary tumor sites the right labium majus, left nasal meatus and nasopharynx, respectively. The other two, one embryonal and one alveolar, were primaries. Grossly, the surgical specimens revealed round or oval, well-demarcated but nonencapsulated masses. Their cut surfaces consisted of homogeneous grayish yellow or white tissue. Microscopically, most tumor cells were poorly differentiated small round, oval or small polygons with eosinophilic cytoplasm. All cases were positive for vimentin, desmin, MyoD1 and myogenin. One embryonal RMS also had a few cells with perinuclear staining of AE1/AE3. The other markers were negative.

CONCLUSIONSAlthough primary or metastatic RMS in breast was almost confined to young adolescent females, our cases suggested that it can also happen to the middle-aged women. Embryonal RMS has a certain metastatic potential. MyoD1 and myogenin are two useful markers when making differential diagnosis. Axillary lymph node status and age may play a role in the prognosis of primary breast RMS patients.

Adolescent ; Adult ; Breast Neoplasms ; diagnosis ; metabolism ; Female ; Humans ; Immunohistochemistry ; Middle Aged ; Rhabdomyosarcoma ; diagnosis ; metabolism ; Young Adult

OBJECTIVETo investigate the optimal operation method for the management of various chronic wounds in legs and feet.

METHODSFifty-one chronic wounds were evaluated according to infection, inflammatory response, and distribution in different areas of the leg and foot. Preoperative treatment was given accordingly, then transposition of skin flap, skin grafting, or amputation was performed. The healing rate after single session operation and average hospitalization were statistically analyzed.

RESULTSThe wound healing rate after single session operation was 86. 3% , the average hospital stay was (17. 8 +/- 2. 1) days, and the appearance and function of the leg and foot after operation was satisfactory.

CONCLUSIONThe appropriate preoperative treatment and operation method conforming to the wound location and evaluation are of vital importance in the management of chronic wounds in the leg and foot. Operation is one of the most effective ways to repair chronic wounds in the leg and foot, and it can shorten the wound healing process and restore the function.

Adult ; Chronic Disease ; Foot Ulcer ; pathology ; surgery ; Humans ; Leg Ulcer ; pathology ; surgery ; Longevity ; Male ; Surgical Flaps ; Wound Healing

In early 2011, the serious outbreak of porcine pseudorabies virus (PRV) infection suddenly recurred in Henan and neighboring Provinces. To investigate the etiology of massive infection with PRV, 16 800 serum samples, 905 porcine epidemic diarrhea virus (PEDV) back-feeding tissues, and 56 PR gene deleted live vaccines were colleted from January 2011 to May 2013 to detect PRV field infection using a PRV gE antibody test kit. The gE and TK genes of 11 new epidemic PRV strains were sequenced by PCR, and their molecular characteristics were analyzed. Moreover, virus titer determination, protective test against PRV, and vaccine potency testing were performed. The results showed that the detection rate of PRV field infection-positive pig farms was 68.06%, and the overall positive rate of PRV field infection in serum was 38.47%; the positive rates in breeding sows, breeding boars, reserve pigs, and commercial pigs were 40.12%, 30.88%, 54.67%, and 26.52%, respectively. The new epidemic strains were in the same evolutionary branch and belonged to the virulent strain group. Compared with the classical PRV strain, the virulence of new epidemic strains changed a little. The length of gE gene was 1 787 bp, and the length of TK gene was 963 bp. The nucleotide homologies of gE and TK genes to Chinese reference strains were 98.2%-99.8% and 98.90%-99.6%, respectively, and the amino acid homologies were 97.1%-99.8% and 97.5%-99.4%, respectively. Commercial vaccine had a 100% protective effect against the new epidemic strains. The positive rate of PRV field infection was 0% in vaccine and 40.44% in back-feeding tissues. The results confirmed that PRV field infection rates were rising sharply among pigs in Henan and neighboring Provinces after 2011. The main virulence genes of new epidemic PRV strains did not change significantly over the years. PR gene deleted live vaccines had no PRV field infection and could completely resist the attack of new strains. The virus carriage of breeding boars and reserve pigs and the serious PRV field infection in PEDV back-feeding tissues were the main causative factors for massive infection with PRV and epidemic outbreak in Henan and neighboring Provinces from 2011 to 2013.
Amino Acid Sequence ; Animal Feed ; analysis ; virology ; Animals ; China ; epidemiology ; Epidemics ; Female ; Herpesvirus 1, Suid ; chemistry ; classification ; genetics ; isolation & purification ; Male ; Molecular Sequence Data ; Phylogeny ; Pseudorabies ; epidemiology ; virology ; Sequence Alignment ; Sequence Homology, Amino Acid ; Sus scrofa ; Swine ; Swine Diseases ; epidemiology ; virology ; Viral Proteins ; chemistry ; genetics

Sterigmatocystin (ST), the secondary metabolite of many kinds of filamentous fungi, is a potent carcinogen structurally related to the aflatoxins (AFT). With similar chemical structure, sterigmatocystion behaves much the homogeneous properties to aflatoxins, both of these mycotoxins exhibit similar biological properties due to their bisfuranoid structure. Since the common, and even heavier pollution, found in foods and feeds-stuff, sterigmatocystion is more harmful than aflatoxins. The reported detection methods of sterigmatocystion included the Thin-layer Chromatography, the High-Performance-Liquid Chromatography, the Enzyme-Linked Immunosorbant Assay and the PCR detection to the toxic gene, however studies about both easy and inexpensive electro-chemical methods have not been found. Our previous studies had discovered that Sterigmatocystin (ST) exist similar sensitivity towards aflatoxin-detoxifizyme (ADTZ), which we had isolated from a fungus, as aflatoxin does. In this work, the preliminary study on electrochemical analysis and determination of ST with triplet electrode enzyme-biosensor system (Ag/AgCl as the reference electrode, Pt and Au as the pair and work electrode, respectively) was carried out. Multiwall-carbon-nanotube (MWNT) had been used to increase the electron transportation on electrode. In the research, the Au electrode was modified by MWNT-immobilized ADTZ, and then the voltammertric behavior of ST was studied by means of cyclic voltammogram analysis and different pulse analysis. Autoprobe CP Research Atomic Force Microscope and TECNAI 10 Transmission Electron Microscope, had been used to detect the MWNT as well as the surface of MWNT-modified ADTZ. The voltammertric behavior of ST was studied by means of cyclic voltammogram analysis and different pulse analysis. The results show that the red-ox peak potential of ST is at the point of -600 mV, the linear detection range is from 8.32 x 10(-5) to 66.56 x 10(-5) mg/mL, the detection limit is at 8.32 x 10(-5) mg/mL, and the response time is 10 seconds. This study provided a good basic work for further research.
Biosensing Techniques ; methods ; Electrochemistry ; Microscopy, Atomic Force ; Microscopy, Electron, Transmission ; Nanotubes, Carbon ; chemistry ; Sterigmatocystin ; analysis

Objective To explore the epileptogenic focus locating methods before and during operation and the curative effect of surgical treatment on the basis of localization. Methods Epileptogenic foci in 621 cases of intractable epilepsy were localized according to clinical manifestations and imaging findings by routine EEG (REEG), ambulatory EEG (AEEG) or video-EEG (VEEG).Different types of operations were performed based on the categories of epilepsy, the preoperatively localized foci and electrocorticogram (ECoG) monitoring results during operations. Results The follow-up ranging 0.5-4 years revealed, according to the criteria of International Antiepileptic Association,that 340 cases(54%)were completely recovered(no postoperative seizure),223 cases(36%)were improved obviously (seizure frequency was decreased by more than 75%), 37 cases (6%) were improved a little(seizure frequency was decreased by 50%-75%)and 21(3%)were not improved(seizure frequency was decreased by less than 50%): the effective rdte was 97%.No case worsened.Conclusions Preoperative integrated localization by EEG principally and intraoperative ECoG monitoring can effectively confirm the site and size of epileptogenic foci to instruct the operation and prevent complications so that the satisfactory treatment efficacy can be achieved.

Objective:To investigate the mechanism of excessive inflammation in the lung of C3H/HeN(C3H) mice following Chlamydia muridarum(Cm) airway infection.Methods:Chlamydial pneumonitis was induced in C3H and C57BL/6(C57) mice by intranasal inoculation with 1×103IFU (inclusion forming unites) of Cm strains.The expression of TLR2,TLR4 and MyD88 mRNA in the lung at different time point post-infection was measured by RT-PCR.Results:Cm infection induced Toll-like receptors expression in two strains of mice.The expression of TLR2 and TLR4 mRNA,especially TLR2 mRNA(P<0.001 or P<0.05),were significantly higher in highly susceptible C3H mice on day 7 and day 14 d post-infection compared with C57 mice.Further studies showed that the expression of MyD88 mRNA was also significantly higher in C3H mice on day 7 post-infection,and maintained high expression untill the day 14.Conclusion:Cm lung infection induced high level of TLR2,TLR4 and MyD88 mRNA expression in C3H mice,which may associate with excessive inflammation in C3H mice.