Acetates ; chemistry ; Aminopyridines ; chemistry ; Angiogenesis Inhibitors ; chemistry ; Benzodiazepinones ; chemistry ; Humans ; Integrin alphaVbeta3 ; antagonists & inhibitors ; chemistry ; Ligands ; Molecular Structure ; Oligopeptides ; chemistry ; Peptides, Cyclic ; chemistry

OBJECTIVEThe SSR information in the transcriptome of Erigeron breviscapus was analyzed in this study, in order to further develop new functional genes SSR markers laid a solid foundation.

METHODSSR loci were searched in all of 52,060 unigenes by using est_timmer. Perl program and SSR primers were designed by Primer3. Furthermore, 36 pairs of primers were randomly selected for the polymorphism analysis on 13 Erigeron breviscapus plants collected from different places.

RESULTA total of 3639 SSRs were found in the transcriptome of Erigeron breviscapus, distributed in 3260 unigenes with the distribution frequency of 6.99%. Di-nucleotide repeat was the main type, account for as much as 34.41% of all SSRs, followed by mono-nucleotide (31.41%) and tri-nucleotide repeat motif (28.08%). The di-nucleotide repeat motifs of AT/AT and AC/GT were the predominant repeat types (28.71%). The tri-nucleotide repeat motifs of AAT/AT was the predominant repeat types (7.94%). For validation the availability of those SSR primers, we randomly selected 36 pairs of primers for PCR amplification. Among them, 34 pair primers (94.44%) produced clear and reproductive bands, 19 pair primers showed polymorphism (52.78%), and 13 Erigeron breviscapus plants were divided into 2 groups.

CONCLUSIONThere are numerous SSRs in Erigeron breviscapus transcriptome with high frequency and various types, this will provide abundant candidate molecular markers for genetic diversity study and genetic map in this plant.

China ; DNA Primers ; genetics ; Erigeron ; classification ; genetics ; Genetic Variation ; Microsatellite Repeats ; Phylogeny ; Polymorphism, Genetic ; Transcriptome

Background Retinitis pigmentosa (RP) is a monogenic inheritance and blinding disease of fundus oculi.There is not an effective therapeutic method now.Objective This work was to identify the mutations of RP1 gene in Chinese RP patients in Ningxia area and to explore the potential interactions in the pathogenesis of RP.Methods The periphery blood of 3-5 ml was collected from 110 individuals with RP(35 ADRP and 75SRP)and 100 normal controls in Ningxia area.Polymerase chain reaction (PCR) and direct DNA sequencing were used to screening the sequence alterations in the entire coding region and splice sites of RP1 gene.Multivariate analysis and two web-based programs( PolyPhen and SIFT) were used to analyze the results.Results Eleven mutation locus were detected in the exon 4 of RP1 gene including two novel sequence variants:p.Lys1152Lys without a higher mutation rate in comparison with normal control group(x2 =9.12 P＜0.01 ),but c.* 247A＞C with a higher mutation rate in comparison with normal control group(x2 =12.77,P＜0.01 ) and c.* 247A＞C mutation was thought to be correlated with RP( r=1.11,P＜0.05 ).The other ten mutation locus were reported as single nucleotide polymorphisms (SNP).The mutation rate of p.Gln1725Gln was found to be higher in the RP patients than the normal controls (x2 =42.09,P＜0.01 ),but no the significant correlation was seen between the pathogenesis of RP and mutation of p.Gln1725Gln(r=1.74,P＞0.05).p.Lys1152Lys mutation was found in only 1 patient.Three SNPs( p.Arg872His,Ala1670Thr,Ser1691Pro) were always occurred in the same 83 RP patient and the relevance ratio was higher than controls ( P＜0.01 ).The age of night blindness on patients with concurrent three mutations was (30.54± 13.68 ) years,and the best corrected visual acuity (BCVA) was 0.50 ± 0.38.The age of night blindness on patients without concurrent three mutations was(21.06± 16.24) years,and the BCVA was 0.40 ±0.33 and were higher than controls ( t =2.11,P ＜ 0.05 ).Conclusions In this study,the prevalence of RP1 mutations among the RP patients in Ningxia population was lower than other populations (＜ 1％ ).The alliance of SNPs (p.Arg872His、p.Ala1670Thr、p.Ser1691Pro) may play a protective role on RP patients and reduce the frequency of mutatiaon in RP1 gene.

In order to generate a pharmacophore model of integrin alphavbeta3 receptor antagonists and design lead compounds which have potent and selective activity against alphavbeta3 receptor with the help of this model. Thirty compounds (four categories) with highly inhibitory activity against the integrin alphavbeta3 receptor (IC50 < 110 nmol x L(-1)), amide, piperazine, piperidine, gamma-valerolactam as the intermediate junction, separately, were selected as a training set to construct a three-dimensional pharmacophore models of integrin alphavbeta3 receptor antagonists with the Catalyst software. The best pharmacophore model of integrin alphavbeta3 receptor antagonists with RMS = 0.73, Correl = 0.90, Weight = 1.17, Config = 14.00 is found out, which consisting of four features: a neg ionizable core (NI), two aliphatic hydrophobic core (HP) and an aromatic ring center (RA). Some new and easily obtained compounds with fine ADME properties and highly potent activity against alphavbeta3 receptor were designed with the new pharmacophore models.
Computer-Aided Design ; Drug Design ; Integrin alphaVbeta3 ; antagonists & inhibitors ; chemistry ; Models, Molecular ; Molecular Structure

Optimization of a process for extracting astaxanthin from Phaffia rhodozyma by acidic method was investigated, regarding several extraction factors such as acids, organic solvents, temperature and time. Fractional factorial design, central composite design and response surface methodology were used to derive a statistically optimal model, which corresponded to the following optimal condition: concentration of lactic acid at 5.55 mol/L, ratio of ethanol to yeast dry weight at 20.25 ml/g, temperature for cell-disruption at 30 degrees C, and extraction time for 3 min. Under this condition, astaxanthin and the total carotenoids could be extracted in amounts of 1294.7 microg/g and 1516.0 microg/g, respectively. This acidic method has advantages such as high extraction efficiency, low chemical toxicity and no special requirement of instruments. Therefore, it might be a more feasible and practical method for industrial practice.
Basidiomycota ; chemistry ; Hydrochloric Acid ; Lactic Acid ; Xanthophylls ; isolation & purification

Objective To investigate the clinical significance of a new method,axis-line-distance technique(ALDT),in scoliosis measurement.Methods Thirty cases with idiopathic scoliosis were measured on two separate occasions by six observers with the Cobb technique and the ALDT on PACS workstation.The interval time between two measurement occasions were three weeks.The data were analyzed statistically with the paired-sample t-test.Results(1)Concerning intraobserver variance in two measurement occasions,the minimum variance,the maximum variance and the average variance were 0, 24.00?,5.71??1.54?for Cobb technique and 0,12.00 mm,(1.95?0.58)mm for ALDT.There were significant measurement differences for four observers with Cobb method 39.00??10.69?versus 36.50?? 10.63?,31.73??10.96?versus 37.30??9.65?,32.03??7.49?versus 27.86??9.00?,29.77??8.87? versus 34.20??7.26?,all P0.05].(2)Concerning interobserver variances in six observers,the average measurement variance was 5.07??0.35?for Cobb method,and(2.32?0.26)mm for ALDT. There were significant measurement differences for every observer using with Cobb method(36.63??10.30? versus 33.27??10.10?,39.00?10.69?versus 31.73??10.96?,32.03??7.49~ versus 29.78?? 8.87?,36.63??10.30?versus 39.00??10.69?versus 32.03??7.48?,39.00??10.69?versus 32.03?? 7.49?,all P

OBJECTIVETo investigate the food water sources of adults in the four cities of China in summer.

METHODSA total of 64 adults aged 18 - 60 yrs from four cities (Beijing, Shanghai, Chengdu and Guangzhou)were selected using convenient sampling method. The food samples were collected by using duplicate portion method and the water content of food samples were determined by using the national standard. The information on amounts and types of daily drinking soup was recorded by subjects using a quantitative measurement.

RESULTSA total of 63 subjects had completed the investigation. The median of daily water from food of subjects was 1157 ml, while water from staple food was 480 ml, the median ratio of which accounting for 41.8% of water from food. Water from dishes was 427 ml, the median ratio of which accounting for 37.9% of water from food. Water from soups was 133 ml, the median ratio of which accounting for 13.8% of water from food. Water from snacks was 0 ml, the median ratio of which accounting for 6.4% of water from food. The medians of water from staple food (384, 388 and 435 ml, respectively) in Beijing, Shanghai and Guangzhou was significant lower than that of Chengdu subjects' (900 ml, χ(2) = 21.27, P = 0.000). But the median proportion of water from staple food of subjects in Beijing (47.7%) was significantly higher than that in Chengdu (43.2%), Shanghai (42.9%) and Guangzhou (33.9%) (χ(2) = 8.69, P = 0.034). The median of amount and proportion of water from soups of subjects in Guangzhou (267 ml, 24.4%) and Chengdu(278 ml, 15.7%) was significantly higher than that in Shanghai (133 ml, 9.0%) and Beijing (100 ml, 5.8%) (amount: χ(2) = 22.52, P = 0.000;proportion: χ(2) = 16.27, P = 0.001).

CONCLUSIONThe staple food and dishes are the main sources of daily food water.

Adolescent ; Adult ; China ; Drinking ; Female ; Food Analysis ; Humans ; Male ; Middle Aged ; Seasons ; Surveys and Questionnaires ; Urban Population ; Young Adult

OBJECTIVETo observe the effects of interleukin-8 monoclonal antibody on smooth muscle cell proliferation and balloon inflation-induced abdominal aorta stenosis in rabbits.

METHODSThirty-six New Zealand white rabbits were randomly assigned to balloon inflation group (group A, n = 12), interleukin-8 monoclonal antibody pre-treated rabbits (2 mg/kg for 3 days before balloon inflation, group B, n = 12) and sham-operated control group (group C, n = 12). Peripheral blood was collected before experiment and at 4 h, 1, 3, 7, 14, and 28 days post balloon inflation or sham operation and the levels of IL-8 were measured by enzyme linked immunosorbent assay (ELISA). The ratio of positive and negative masculine cells in the high power microscopic field was determined in proliferating cell nuclear antigen (PCNA) stained slide. Histopathologic examination was performed in abdominal aorta and luminal area, intima and tunica media area were measured.

RESULTSPlasma interleukin-8 began to rise at 4 h and peaked at 1 day and remained increased up to 28 days after balloon inflation in rabbits of group A, plasma interleukin-8 level in group A was significantly higher than in group B and C at 4 h and thereafter post operation. The ratio of positive and negative masculine cells was significantly increased in group A compared to group C and was significantly lower in group B than in group A. Abdominal aorta stenosis, luminal area, intima and tunica media area were significantly reduced in group B than in group A. Correlation analysis indicated that there were positive relations between plasma IL-8 level and intima thickness, area of intima and tunica media, respectively (r = 0.894, 0.783, 0.801, 0.912, all P < 0.01).

CONCLUSIONSPlasma IL-8 level is increased in this abdominal aorta stenosis model and is positively correlated to the severity of abdominal aorta stenosis. IL-8 monoclonal antibody could significantly reduce abdominal aorta stenosis in this abdominal aorta stenosis model.

Animals ; Antibodies, Monoclonal ; pharmacology ; therapeutic use ; Aorta, Abdominal ; pathology ; Aortic Coarctation ; drug therapy ; pathology ; Cell Proliferation ; drug effects ; Interleukin-8 ; immunology ; Muscle, Smooth, Vascular ; cytology ; drug effects ; Myocytes, Smooth Muscle ; drug effects ; Rabbits

Fermentation of Phaffia rhodozyma is a major method for producing astaxanthin, an important pigment with industrial and pharmaceutical application. To improve astaxanthin productivity, single factor and mixture design experiments were used to investigate the effects of nitrogen source on Phaffia rhodozyma cultivation and astaxanthin production. Results of single factor experiments showed nitrogen source could significantly affect P. rhodozyma cultivation with respect to carbon source utilization, yeast growth and astaxanthin accumulation. Further studies of mixture design experiments using (NH(4))(2)SO(4), KNO(3) and beef extract as nitrogen sources indicated that the proportion of three nitrogen sources was very important to astaxanthin production. Validation experiments showed that the optimal nitrogen source was composed of 0.28 g/L (NH(4))(2)SO(4), 0.49 g/L KNO(3) and 1.19 g/L beef extract. The kinetic characteristics of batch cultivation were investigated in a 5-L pH-stat fermentor. The maximum amount of biomass and highest astaxanthin yield in terms of volume and in terms of biomass were 7.71 mg/L and 1.00 mg/g, respectively.
Bioreactors ; microbiology ; Cell Culture Techniques ; methods ; Cell Proliferation ; Computer Simulation ; Mitosporic Fungi ; physiology ; Models, Biological ; Nitrogen ; metabolism ; Xanthophylls ; biosynthesis

PURPOSE: Insufficient sensitivity and specificity prevent the use of most existing biomarkers for early detection of breast cancer. Recently, it was reported that serum microRNAs (miRNAs) may be potential biomarkers in many cancer diseases. In this study, we investigated whether serum levels of 5 miRNAs including miR-21, miR-125b, miR-145, miR-155, and miR-365 could discriminate breast cancer patients and healthy controls. METHODS: Serum levels of miRNAs were measured by using quantitative real-time polymerase chain reaction in 99 breast cancer patients and 21 healthy controls. The abundance change of serum miRNAs were also evaluated following surgical resection in 20 breast cancer patients. Receiver operating characteristic (ROC) curve analysis was performed to assess the sensitivity and specificity of miRNAs as diagnostic biomarkers. RESULTS: Serum levels of miR-21 and miR-155 was significantly higher, while miR-365 was significantly lower in breast cancer as compared with healthy controls. The serum levels of miR-21 and miR-155 significantly decreased following surgical resection. Additionally, the serum level of miR-155 at stages I and II was significantly higher compared to stage III. The serum miR-145 level was remarkably higher in progesterone receptor (PR)-positive patients than PR-negative. The positivity of miR-21, miR-155, and miR-365 was high compared to CA 153 and CEA in breast cancer. ROC curve analyses of a combination of miR-21, miR-155, and miR-365 yielded much higher area under curve and enhanced sensitivity and specificity in comparison to each miRNA alone. CONCLUSION: The combination of serum miR-21/miR-155/miR-365 may potentially serve as a sensitive and specific biomarker that enables differentiation of breast cancer from healthy controls.
Area Under Curve ; Biomarkers ; Breast Neoplasms* ; Breast* ; Humans ; MicroRNAs ; Real-Time Polymerase Chain Reaction ; Receptors, Progesterone ; ROC Curve ; Sensitivity and Specificity