Humans ; Laryngeal Neoplasms ; Male ; Middle Aged ; Neoplasms, Muscle Tissue

Objective To study the receptors for the axon guidance factor Netrin-4 and whether the known Netrin-1 receptors, DCC and UNC5H1, are coreceptors for Netrin-4. Methods The receptor for Netrin-4 was detected by affinitycytochemistry and in situ staining with the alkaline phosphatase (AP) tagged Netrin-4 fusion protein. The dynamic character of the binding between Netrin-4 and its receptor was evaluated through ligand-receptor binding assay and competitive binding assay. Results In situ staining of AP4-Netrin-4 protein bound to COS7 cells transiently transfected with eukaryotic expression coding DCC and UNC5H1 showed that the Netrin-4 could bind to the transfected cells. Competitive binding assays showed that Netrin-1 could compete with Netrin-4 for the same receptors. Conclusion DCC and UNC5H1 are coreceptors for Netrin-4. However, because the binding affinity of Netrin-4 with its receptor was lower than that of Netrin-1,it could be inferred that there should exist unique receptor(s) for Netrin-4 other than DCC and UNC5H1.

To establish a molecular identification method for Bletillae Rhizoma, this paper extracted genome DNA from Bletillae Rhizoma and its adulterants. The sequences of rDNA ITS2 were sequenced after amplifying. Then multiple alignments of ITS2 were constructed phylogenetic tree with Neighbor Joining by MEGA 5. 1 and found out SNPs loci. The result showed that rDNA ITS2 region could identify Bletillae Rhizoma and its adulterants. There existed the SNPs loci, which could identify Bletilla striata and B. ochracea. Furthermore, we designed specific primers against the SNPs loci of B. striata and B. ochracea, then screened primers and optimized the PCR amplification conditions. Finally, the DNA of B. striata and B. ochracea were specifically amplified by BJ59-412F, BJ59-412R and HHBJ-225R. The length of amplification products were respectively about 350 bp and 520 bp that were effectively identified of B. striata and B. ochracea. While, the adulterants of Bletillae Rhizoma were no-reaction occurring. To sum up, the amplification conditions of the primers can identify B. striata, B. ochracea and their adulterants successfully at the same time. This method was easy, time-saving, and reliable, which can be used as a rapid method for molecular identification of Bletillae Rhizoma.
Base Sequence ; DNA Primers ; genetics ; DNA, Intergenic ; genetics ; DNA, Plant ; genetics ; Drug Contamination ; prevention & control ; Molecular Sequence Data ; Orchidaceae ; classification ; genetics ; Phylogeny ; Polymorphism, Single Nucleotide ; Rhizome ; classification ; genetics

To establish a convenient and rapid method for identification of Pseudostellariae Radix by molecular identification, the rDNA-ITS sequences of Pseudostella riaheterophylla and its adulterants had been aligned to find out specific fragment. The specific primers against the fragment were designed and the PCR amplification conditions were optimized. The fluorescence reaction of the PCR products colored by 100 x SYBR Green I was observed under UV. The concentration of reaction buffer included 5.5 μL DNA Taq polymerase premix, 10 pmol Tzs-2F and 10 pmol Tzs-2R, 20-80 ng template DNA, and plus double sterile distilled water to 25 μL. The PCR thermal profile was as follows: predenaturation at 95 degrees C for 1 min, followed by 30 cycles of denaturation at 95 degrees C for 5 seconds, primer annealing and extension at 56 degrees C for 15 seconds, then it was extension at 72 degrees C for 30 seconds. The fluorescence reaction of Pseudostellariae Radix showed green fluorescence, while adulterants had not. Extraction, amplification DNA and all steps of molecular identification could be completed successfully in 40 minutes. The approach could amplify DNA template of Pseudostellariae Radix specificity, and its product with 1 μL 100 x SYBR Green I could engender green fluorescence under UV. The method was simple and accurate, so it could be used for identification of Chinese traditional medicine.
Caryophyllaceae ; classification ; genetics ; DNA Primers ; genetics ; Drug Contamination ; prevention & control ; Plant Roots ; classification ; genetics ; Polymerase Chain Reaction ; methods

Objective Neuropsychiatric systemic lupus erythematosus ( SLE) is a common complication of SLE, whose path-ogenesis is not yet clear but associated with the alteration of cerebral blood flow ( CBF) in some studies.This study was to investigate the CBF alteration in SLE patients without overt neuropsychiatric symptoms by arterial spin labeling ( ASL) MRI. Methods Twenty-eight SLE patients without overt neuropsychiatric symptoms and 30 age-and sex-matched healthy controls underwent conventional MRI and ASL examinations, and all received such neuropsychologic tests as number connecting test-A ( NCT-A ) , digit symbol test ( DST ) , self-rating anxiety scale ( SAS ) , and self-rating depression scale ( SDS) .Independent sample-t test was used to detect the mean CBF in the whole brain, gray matter, and white matter of the SLE patients and healthy controls.The voxel-wise CBF maps of the two groups of subjects were further analyzed with the SPM8 software to compare the regional CBF between the two groups, followed by evaluation of the correlation between the regional CBF values and clinical markers. Results In comparison with the healthy controls, the SLE pa-tients showed significantly reduced CBF in the gray matter (40.5 ±3.7 vs 37.3 ±6.5, P=0.028) and the whole brain (38.0 ±3.5 vs 35.1 ±6.1, P=0.032), especially in the supplementary motor area and the adjacent middle cingulate, anterior cingulate, left medial frontal gyrus, left inferior frontal gyrus, and left insula (P<0.05, FWE corrected).The NCT-A score was negatively correlated with the CBF values of the left medial frontal gyrus (r=-0.402, P=0.032) and left inferior frontal gyrus (r=-0.382, P=0.045) of the SLE patients. Conclusion ASL and MRI showed significantly reduced cerebral blood flow in the SLE patient without overt neu-ropsychiatric manifestations, which was correlated with the change of the patient's cognitive function.

Objective To explore the prognostic value of biomarkers in type 2 diabetic patients with acute myocardial infarction (AMI), this study was to investigate the associations between the neutrophil to lymphocyte ratio (NLR), the Global Registry of Acute Coronary Events (GRACE) score and in-hospital mortality. MethodsSeven hundred and seven consecutive AMI patients were divided into diabetic group (DM-AMI group), impaired glucose tolerance group (IGT-AMI group), and normal glycemic group (NGT-AMI group). The laboratory and clinical characteristics were assessed retrospectively from the medical records. The NLR and GRACE score were calculated. Results In AMI patients, the DM-AMI group had significantly higher NLR and GRACE scores compared with those from the IGT-AMI group and NGT-AMI group (P<0.01 or P<0.05). In DM-AMI group, the NLR and GRACE score were considerably elevated in the elderly DM-AMI group compared with their younger counterparts (both P<0.01). Furthermore, the NLR was considerably higher in the high-risk group than those in both the low- and medium-risk groups based on the GRACE score (both P<0.01). The NLR was positively correlated with the GRACE score in DM-AMI group(r=0.425, P<0.01). The NLR level and GRACE score were higher in the death group than those in surviving patients (both P<0.01). The optimal cut-off levels of 9.36 for NLR and 166 for GRACE score seem to predicte death in-hospital. Based on the receiver operating characteristic curve, when to predict death in-hospital, the best cutoff value of NLR was 9.36 (sensitivity 80.8%, specificity 69.6%; area under curve 0.787), and the best cutoff value of GRACE score was 166 (sensitivity 76.9%, specificity 76.4%; area under curve 0.778). Conclusion An elevated NLR is a potential predictor of in-hospital mortality in type 2 diabetic patients with AMI, which could help clinicians indentify high-risk patients and determine appropriate treatment strategies. <英文关鍵词>>=Neutrophil to lymphocyte ratio; In-hospital mortality; Acute myocardial infarction; Diabetes mellitus, type 2

Referring to the rules for agricultural seed testing (GB /T 3543-1995) issued by China, the test of sampling, seed purity, weight per 1 000 seeds, seed moisture, seed viability and germination rate had been studied for screening seed quality test methods of Pesudostellaria heterophylla. The seed quality from different collection areas was measured. The results showed that at least 6.5 g seeds should be sampled and passed through 10-mesh sieve for purity analysis. The weight of 1 000 seeds was determined by using the 500-seed method. The phenotypic observation and size measurement were used for authenticity testing. The seed moisture was determined under the higher temperature (130 ± 2) degrees C for 5 hours. The seeds were dipped into 0.2% TTC sustaining 1 hour at 40 degrees C, then the viability could be determined. The break dormancy seeds were cultured on sand at 10 degrees C. K cluster analysis was applied for the data analysis, the seed quality from different collection areas grading of P. Heterophylla was described as three grades. The seed quality of each grade should reach following requirements: for first grade seeds, germination rate ≥ 86%, 1 000-grain weight ≥ 2.59 g, purity ≥ 87%, moisture ≤ 13.1%; for second grade seeds, germination rate ≥ 70%, 1 000-grain weight ≥ 2.40 g, purity ≥ 77%, moisture ≤ 14.3%; for third grade seeds, germination rate ≥ 41%, 1 000-grain weight ≥ 2.29 g, purity ≥ 76%, moisture ≤ 15.8%. The seed testing methods for quality items of P. heterophylla had been initially established, as well as the primary P. heterophylla seed quality classification standard.
Caryophyllaceae ; chemistry ; growth & development ; Germination ; Quality Control ; Seeds ; chemistry ; growth & development ; Water ; analysis

Objective To analyze the present situation of the dental instrument cleaning and disinfection in Hainan province and to explore the management mode of dental instrument cleaning and disinfection which is suitable for the provincial situation .Meth-ods By adopting the stratified random sampling method according to the hospital grades ,24 hospitals in Hainan province were per-formed the questionnaire survey on the cleaning mode of oral instruments ,layout of cleaning and disinfection room ,cleaning method and facilities ,protection measures and training of cleaning staff .Results 14 hospitals (58 .33% ) had the sterilization and supply center for conducting the centralized processing on the dental instruments .The tertiary hospitals and the second-grade hospitals had the independent cleaning and disinfection rooms with the rational layout and professional cleaning staff ;the safeguard facilities had the application in place ,the training of the related cleaning and disinfection work and the cleaning process conformed the require-ment of the standards .Among 10 first-grade and below hospitals ,only 1 hospital(10 .00% ) had the rational layout of cleaning and disinfection rooms ;3 hospitals(30 .00% ) had the professional cleaning staff ;the related training of the cleaning staff was not basi-cally carried out and the safeguard was not in place ,most of the cleaning and disinfection instruments and the cleaning process were not in accordance with the requirements .The qualification rates of instruments cleaning and disinfection in different grades of hospi-tals by the ATP bioluminescence assay were 100 .00% ,90 .00% and 80 .00% .Conclusion The existing problems are general and prominent in the hospitals of the first-grade and below .It is suggested that the regionalized disinfection and supply management mode is implemented for maximally realizing the optimized resource configuration in the disinfection and supply center .

OBJECTIVETo compare the ability of osthole (OST) and genistein (GEN) in enhancing bone peak bone mass of rats to prevent osteoporosis.

METHODSThirty-six female one-month-old SD rats of (125 +/- 3) g body weight were randomly divided into three groups, 12 rats in each group, one group was orally administered osthole at 9 mg x kg(-1) d(-1), one group was given genistein at 10 mg x kg(-1) d(-1) and another was given equal quantity of distilled water as the control. The body weight was monitored weekly and the bone mineral density (BMD) of total body was measured every month. All rats were sacrificed after three months, the femoral bone mineral density, the serum levels of osteocalcin (OC) and anti-tartaric acid phosphatase 5b (TRACP 5b) were measured by Elisa. The bone microarchitectures were analyzed with micro-CT and the bone biomechanics properties were tested with universal material machine.

RESULTSNo significant differences were observed between O-treated or GEN group and the control for the food-intake and body weight during three months. However, the rats treated with OST had significant higher BMD for both total body and femur than the control and GEN group. The O-treated rats also had higher level of serum OC and lower level of TRACP 5b. Besides, they owned bigger bone volume/tissue volume, trabecular thickness, trabecular number but smaller trabecular spacing. In the three point bending tests of femurs,they were found to have larger maximum load, the young's modulus and structural model index (SMI).

CONCLUSIONOrally administered osthole could efficiently increase the peak bone mass of rats,which provide new ideas for preventing osteoporosis.

Acid Phosphatase ; blood ; Animals ; Body Weight ; drug effects ; Bone Density ; drug effects ; Coumarins ; pharmacology ; Female ; Femur ; diagnostic imaging ; drug effects ; pathology ; Genistein ; pharmacology ; Isoenzymes ; blood ; Osteocalcin ; blood ; Radiography ; Rats ; Rats, Sprague-Dawley ; Tartrate-Resistant Acid Phosphatase

OBJECTIVETo evaluate effect of amygdalin on expression of four biomarkers in the animal model of pulmonary fibrosis induced by bleomycin.

METHODSRats were given one dose (5 mg/kg) of bleomycin in bleomycin-treated groups, amygdalin-treated groups and saline in controls by intratracheal instillation exposed surgically. The amygdalin-treated groups rats were treated with intraperitoneal injection of amygdalin (15 mg x kg(-1) x day(-1)). The rats were sacrificed 7, 14 and 28 days after bleomycin administration. Polarized light microscopy and Image-Pro Plus detected I and III collagen expressed in Paraffin-embedded lung sections stained with Sirius red. Surface-enhanced laser desorption-ionization time-of-flight mass spectrometry (SELDI-TOF MS) with weak cationic proteinchip (CM10) detected differentially expressed proteins in the pooled serum samples of all groups.

RESULTSConsistent fibrotic responses were found in all bleomycin and amygdalin-tread groups. On the 7th, 14th and 28th day after bleomycin or saline instillation, four differentially expressed proteins were detected in the pooled serum of all groups rats, consisting of 4 proteins with mass/charge ratio of 3530.7, 7043.5, 8332.6 and 9068.0, respectively. Compared with control groups, protein peaks intensity ratio with mass/charge ratio of 3530.7 on 7, 28 d and 7043.5, 8332.6 and 9068.0 on 7, 14 and 28 d was > 2 in bleomycin-treated groups. Compared with amygdalin-treated groups, protein peaks intensity with mass/charge ratio of 3530.7 at 7, 14, 28 d had no change almost, but protein peaks intensity ratio with mass/charge ratio of 7043.5 at 7 d, 8332.6 on 28 d and 9068.0 on 14 d was > 2 in bleomycin-tread groups. All the four protein peaks intensity had no change almost at other point.

CONCLUSIONAmygdalin may reduce the bleomycin-induced increase of differentially expressed protein peak intensities in rat serum.

Amygdalin ; pharmacology ; Animals ; Biomarkers ; blood ; Bleomycin ; adverse effects ; Blood Proteins ; metabolism ; Male ; Pulmonary Fibrosis ; blood ; chemically induced ; Rats ; Rats, Wistar