OBJECTIVETo compare the tissue engineered cartilage constructed with chondrocytes derived from auricular and articular cartilage.

METHODSChondrocytes were isolated from porcine auricular and articular cartilage, and BMSCs were obtained from bone marrow aspirate and cultured. Each kind of chondrocytes were resuspended alone or mixed with BMSCs at a ratio of 1:1, and seeded onto PGA/PLA scaffolds to construct tissue engineered cartilage (n = 4). The constructs were cultured for 8 weeks in vitro and then subcutaneously implanted into nude mice for 6 weeks. The differences between chondrocytes monoculture from articular and auricular cartilage or between each of them co-cultured with BMSCs were evaluated by gross view, measurement of thickness and wet weight, histological examinations including H&E, Safranin O, type II collagen, and Ponceau's & Victoria blue staining, and gene expression analysis of cartilage related genes.

RESULTSNo obvious differences were found histologically among the complexes constructed in vitro 8 weeks except for few elastic fibers secreted in the auricular chondrocytes + BMSCs co-culture group. Neo-cartilage is thicker in the groups of articular chondrocytes (38. 1% than the group of auricular chondrocytes, P < 0.05) and articular chondrocytes + BMSCs co-culture (19.3% than the group of auricular chondrocytes + BMSCs, P < 0.05). However, after 6 weeks in vivo the elastic fibers were found positive in the complexes constructed by auricular chondrocytes, and its staining was even stronger and more homogenous in the group of auricular chondrocytes + BMSCs co-culture. The tissues generated by articular chondrocytes alone and co-cultured with BMSCs both formed the characteristic features of three-layer structure of hyaline cartilage and ossified in vivo with significant up-regulation of COL10A1 and MMP-13. To summarize, auricular chondrocytes formed the elastic cartilage while articular chondrocytes formed the hyaline cartilage during the development of tissue engineered cartilage either by monoculture or the co-culture with BMSCs.

CONCLUSIONSThe chondrogenic response of chondrocytes from different cartilage origins demonstrates that an initial chondrocyte and cartilage type recapitulates the same in later tissue-engineered development.

Animals ; Bone Marrow Cells ; cytology ; Cartilage, Articular ; cytology ; Cells, Cultured ; Chondrocytes ; Coculture Techniques ; Ear Auricle ; cytology ; Mesenchymal Stromal Cells ; cytology ; Mice, Nude ; Swine ; Tissue Engineering ; methods ; Tissue Scaffolds

Objective:To establish an HPLC method to determine vanillin in 18 dairy products. Methods:The samples were sep-arated on a C18 column (250 mm × 4. 6 mm, 5 μm) with the mobile phase of methanol and water (35∶65) at the flow rate of 1. 0 ml ·min-1 . The detection wavelength was 275 nm. Results:The LOD ( limit of detection) of vanillin was 0. 45 μg·ml-1 . Excellent linearity was obtained within the concentration range of 1.990-99.500 μg·ml-1(r=1.000 0). The average recovery of vanillin in different dairy products with low, medium and high levels varied between 96. 26% and 100. 81% with RSD of 0. 15%-2. 22%. Con-clusion:The method is simple and reproducible, which can be applied in the rapid analysis of vanillin in dairy products.

Objective:To investigate the effect of the ceramometal b ridge on the local gingival groove flora.Methods: Classi cal bacterial incubation and identification were used to study the changes of th e local gingival groove flora in 6 patients wearing ceramometal bridge for 1 wee k to 3 months. Results: 3 months after the prosthetic procedure the CFU o f P.melaninogenica, Gram-positive bacilli cmainly Actinomyces and the t otal bacteria were significantly increased (P

Objective: To evaluate the testing level of the relevant items in food inspection agency laboratory objectively, analyze the existing problems and help them enhance the ability of detection.Methods: The blind sample assessment of sodium saccharin in drinks was organized and performed.The test samples at high and low concentrations were prepared and studied by the tests of homogeneity and stability.Those met the requirements of blind sample assessment were randomly distributed to 193 laboratories and the returned data were analyzed statistically.Results: Totally 182 valid data were collected.Among the reported results, those from 85 laboratories were satisfied in the low concentration group with the satisfaction rate of 87.6% , those from 12 laboratories were not satisfied, and no results were suspicious;as for the high concentration group, the above data was 67(78.8%), 12 and 6, respectively.The overall satisfaction rate was 83.5%.The results, especially the dissatisfied and suspicious data, were analyzed by the original records of each participating laboratory, the analysis focused on 8 aspects including detection method, instrument brand, control product variety, pretreatment method, recovery rate, column selection and the other influencing factors, and suggestions were given to every laboratory to improve its own situation.Conclusion: The statistical data reflect that our food testing laboratory has strong detection ability in the determination of saccharin sodium in beverages, and can provide powerful technical support for the regulatory authorities.

Objective To observe the effectiveness of introducing Diazepam into combined aesthesia of Sumianxin and ketamine hydrochloride .Method A total of 80 rabbits of both genders for operation were randomly divided into A , B and C groups .The A group was injected with Sumianxin intramuscularly ( 0.3 mL/kg by weight ) .The B group was injected with Sumianxin and ketamine hydrochloride intramuscularly ( 0.3 mL/kg by weight ) .The C group was injected with Diazepam intravenously ( 1.5 mL/kg by weight ) combined with Sumianxin and ketamine hydrochloride injected intramuscularly (0.3 mL/kg by weight).The aesthetic effects, induction time, anesthesia maintaining time, total anaesthetic dose and operation time were observed , recorded and compared .Result The induction time of the C group was significantly shorter than A and B groups (P<0.01).The initial anesthesia maintaining time of the C group was the longest among the three (P<0.01) with least total anaesthetic dose (P<0.01).The operation time of the C group was the least with best aesthetic effects (P<0.01).Conclusion Introducing Diazepam into combined aesthesia of Sumianxin and ketamine hydrochloride can improve the aesthetic effects .Therefore , this is an optional aesthetic method for time-consuming animal operation or sensitive surgical sites of rabbits .

Objective To isolate Fusarium species from Kashin-Beck disease(KBD)area and biosynthesize cnlde T-2 toxin.Methods T-2 toxin.producing Fusarium was isolated from corns produced in KBD area and purifted.The purifted funsi were identified according to the traits of colony,appearance of thallus and characters of conidium and then weIe cultivated in sterile Corn culture media.After extraction with organic solvent and purification by silica gel chromatography column,the quality and quantity of the toxin in the extracts were estimated by thin,Layer chromatography and high-performance liquid chromatography.Results The toxin-producing strain was Fusarium tricinctum. The com cuIture media inoculated with this strain produced about 250 mg of crude T-2 toxin per kg. Conclusions This experiment has indirectly further confirmed pollution of T-2 toxin-producing Fmarium existed in

PAK1 plays an important role in the development of tumors. It is of great significance to screen and develop new PAK1 inhibitors as targeted drugs for cancer treatment. The traditional PAK1 inhibitor screening method has the problems of high cost and low efficiency. Computer virtual screening can reduce the cost of finding active lead compounds and improve the screening efficiency. In this study, a kind of PAK1 candidate compound was screened by computer assisted virtual screening combined with Z′lyteTM high flux kinase screen. In vitro enzyme activity screening showed that compound 18(K788)had good PAK1 inhibitory activity(inhibition rate was 42. 7%). Furtherly by MTT detection, it was found that K788 had significant PAK1 positive tumor killing activity, which was even better than the positive drug IPA-3. Flow cytometry and Western Blot showed that K788 could activate caspase apoptosis pathway and induce apoptosis of colon cancer cell DLD-1 by inhibiting PAK1 expression and activation. K788 has great potential for clinical development and application, and can be used as a PAK1 target for further research.

Objective To analyze the pathogenic epidemiological characteristics of acute lower respiratory tract infection(ALRTI) in children in Gansu Province from 2012 to 2015. Methods The surveillance data of 458 children infected with ALRTI in 10 sentinel hospitals in Gansu province from 2012 to 2015 were collected, and infection status and epidemiological characteristics of each virus and bacteria were analyzed by descriptive study methods. Results The male to female ratio of the 458 children with ALRTI was 1.81:1, and the positive detection rate of the virus was 33.62%（95% CI：29.28%-37.97%), among which the positive detection rate of respiratory syncytial virus was the highest (12.23%).The positive detection rate of bacteria was 24.84%（95% CI：20.04%-29.65%),among which the positive detection rate of streptococcus pneumoniae was the highest (18.47%).There was significant no difference in the positive detection rate of virus and bacteria between children of different genders (P>0.05).There was significant no difference in the positive detection rate of virus among children of different age groups ( 2=5.980,P=0.050), but the positive detection rate of bacteria was different ( 2=12.078,P=0.002).Positive detection rates of virus infection and bacterial infection were different in distinct seasons (all P<0.05). By using logistics regression analysis, season, age and sentinel hospital were the influencing factors of ALRTI virus infection in children (all P<0.05), and sentinel hospital and years were the influencing factors of ALRTI bacterial infection in children (all P<0.05). Conclusion Respiratory syncytial virus, influenza virus and parainfluenza were the main causes of ALRTI virus infection in children aged 0-14 years in Gansu province, and the main bacterial infections were streptococcus pneumoniae and haemophilus influenzae, the number of virus infection was more than that of bacterial infection. Viral and bacterial infection had the same peak incidence.

OBJECTIVETo investigate the mechanism of polymyxin B (PMB) antagonizing the biological activity of lipopolysaccharide (LPS).

METHODSThe affinity of PMB for LPS and lipid A was assayed by biosensor, and the neutralization of PMB for LPS (2 ng/ml) was detected by kinetic turbidimetric limulus test. The releases of TNF-alpha and IL-6 in murine peritoneal macrophages a (PMphi) after exposure to LPS (100 ng/ml) were detected, and the expression levels of TLR4, TNF-alpha and IL-6 mRNA in PMphi induced by LPS (100 ng/ml) were measured by RT-PCR.

RESULTSPMB had high-affinity to LPS and lipid A with dissociation equilibrium constants of 18.9 nmol/L and 11.1 nmol/L, respectively, and neutralized LPS in a dose-dependent manner. Furthermore, PMB could markedly inhibit the expressions of TLR4, TNF-alpha and IL-6 mRNA and the release of cycokines in LPS-stimulated murine peritoneal macrophages.

CONCLUSIONSPMB neutralizes LPS and inhibites the expression and release of cycokines in macrophages, in which the affinity of PMB for lipid A plays an important role.

Animals ; Cytokines ; analysis ; Limulus Test ; Lipid A ; antagonists & inhibitors ; Lipopolysaccharides ; antagonists & inhibitors ; Macrophages ; chemistry ; Mice ; Polymyxin B ; pharmacology

OBJECTIVETo explore the role of autophagy in quercetin (Que)-induced apoptosis in human bladder carcinoma BIU-87 cells in vitro.

METHODSTo determine the proliferative inhibition by MTT colorimetric assay after treating BIU-87 cells with quercetin at various concentrations. To identify autophagy and apoptosis in the BIU-87 cells after Que treatment by monodansylcadaverin (MDC) and Hoechst 33258 fluorescent staining, respectively. To examine the cytotoxic effect of Que and influence of autophagy on apoptosis by studying LDH leakage rate and flow cytometry, after blocking the autophagy with 3-methlyadenine (3-MA), a specific autophagy inhibitor.

RESULTSThere was an obvious inhibitory effect of Que on the proliferation of BIU-87 cells in a time- and dose-dependent manner. The inhibition rate of BIU-87 cells after 200 µmol/L Que treatment for 72 hours was 89.2%. Autophagy and apoptosis were induced and detected in Que-treated BIU-87 cells and autophagy occurred earlier than apoptosis. The apoptosis peak became much higher after the autophagy was blocked. Whenever the autophagy was blocked before or after Que treatment, the Que-induced cytotoxicity in BIU-87 cells was enhanced.

CONCLUSIONSQuercetin significantly inhibits the proliferation of BIU-87 cells, and the autophagy is induced earlier than apoptosis. In the process of Que-induced apoptosis of BIU-87 cells, autophagy may play a protective role at the initiation phase, delay apoptosis and reduce the Que-induced death of BIU-87 cells.

Adenine ; analogs & derivatives ; pharmacology ; Antioxidants ; administration & dosage ; pharmacology ; Apoptosis ; drug effects ; Autophagy ; drug effects ; physiology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Dose-Response Relationship, Drug ; Humans ; L-Lactate Dehydrogenase ; drug effects ; metabolism ; Quercetin ; administration & dosage ; pharmacology ; Urinary Bladder Neoplasms ; pathology