Screen small molecule peptide specific binding to small cell lung cancer cell (DMS53) was screened by using the "one-bead one-peptide" combinatorial technology. Thirty two positive beads binding to DMS53 were totally obtained after primary screening. Consensus peptide sequences of cXNGRXXc and cNGRXXXc were identified by amino acid sequencing in ten beads. Three representative peptides were re-synthesized on beads. Secondary screening showed that cell adhesion percentage of cFNGRQQc to DMSS was higher than the other two peptides. cFNGRQQc was further studied for cell specificity, alanine scanning and site-directed deletion. The results showed that cFNGRQQc is specific for promoting cell adhesion to DMS53 but not to other human cell lines. Both motif of -NGR- and the length of six peptide of cFNGRQQc structure are important for DMS53 attachment. In an antibody or peptide blocking assay, cell adhesion of DMS53 to peptide bead was not inhibited by antibodies or peptides including anti-integrin, E-cadherin, NCAM and ICAM. The binding site on DMS53 surface for cFNGRQQc peptide need to be proven in the future.

Monascus spp.,a kind of filamentous fungi,produce abundant of important metabolites which were widely used in the fields of food and medicine.Until now,there are few reports on the important functional genes of the Monascus spp.due to little genetic information.In this paper,the feasibility of gene deletion mediated via Agrobacterium tumefaciens on the basis of homologous recombination was analyzed by studying on the deletion of the RGS domain of putative G-protein signaling regulator gene mrfA in Monascus ruber.The length of homologous arms of deletion vector pC805S were 958 bp and 824 bp,respectively.There were 26 transformants in which homologous recombination occurred in 138 transformants and the recombination rate was 18.8%.The result showed it was feasible to identify the function of unknown gene in M.ruber with the targeted-deletion technology mediated via A.tumefaciens.

OBJECTIVETo screen out main molecular target promoting human neural stem cells (NSCs) of ginsenoside Rg1 by using the gene chip technology.

METHODFirst, MTT assay was adopted to screen out the optimal concentration of Rg1-promoted NSC proliferation (120 mg x L(-1)). Then, on the 7th day after the Rg1-promoted NSC proliferation, the expression of target genes was observed by the gene chip technology. The most important target gene and signal transduction pathways were screened out through the data calculations.

RESULTOn the 7th day after the Rg1-promoted NSC proliferation, obtained 440 differential genes, 266 significantly upregulated genes and 174 significantly down-regulated genes. HES1 gene, CAMP (cyclic adenosine monophosphate)-PKA (protein kinase A) and PI3K (phosphatidylinositol 3 kinase)-AKT signal transduction pathways were closely related to the NSC proliferation.

CONCLUSIONThe differentially expressed genes screened out by gene chip may provide new clues for studies on molecular mechanism of ginsenoside Rg1-promoted NSCs proliferation.

Cell Proliferation ; drug effects ; Ginsenosides ; pharmacology ; Humans ; Neural Stem Cells ; cytology ; drug effects ; metabolism ; Oligonucleotide Array Sequence Analysis ; RNA ; genetics ; isolation & purification

BACKGROUNDPreviously we had successfully tracked adult human neural stem cells (NSCs) labeled with superparamagnetic iron oxide particles (SPIOs) in host human brain after transplantation in vivo non-invasively by magnetic resonance imaging (MRI). However, the function of the transplanted NSCs could not be evaluated by the method. In the study, we applied manganese-enhanced MRI (ME-MRI) to detect NSCs function after implantation in brain of rats with traumatic brain injury (TBI) in vivo.

METHODSTotally 40 TBI rats were randomly divided into 4 groups with 10 rats in each group. In group 1, the TBI rats did not receive NSCs transplantation. MnCl2·4H2O was intravenously injected, hyperosmolar mannitol was delivered to disrupt rightside blood brain barrier, and its contralateral forepaw was electrically stimulated. In group 2, the TBI rats received NSCs (labeled with SPIO) transplantation, and the ME-MRI procedure was same to group 1. In group 3, the TBI rats received NSCs (labeled with SPIO) transplantation, and the ME-MRI procedure was same to group 1, but diltiazem was introduced during the electrical stimulation period. In group 4, the TBI rats received phosphate buffered saline (PBS) injection, and the ME-MRI procedure was same to group 1.

RESULTSHyperintense signals were detected by ME-MRI in the cortex areas associated with somatosensory in TBI rats of group 2. These signals, which could not be induced in TBI rats of groups 1 and 4, disappeared when diltiazem was introduced in TBI rats of group 3.

CONCLUSIONIn this initial study, we mapped implanted NSCs activity and its functional participation within local brain area in TBI rats by ME-MRI technique, paving the way for further pre-clinical research.

Animals ; Brain Injuries ; physiopathology ; surgery ; Cell Movement ; Image Enhancement ; Magnetic Resonance Imaging ; methods ; Manganese ; Neural Stem Cells ; physiology ; transplantation ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo study the infection status of Leptospira in rodents on Heixiazi island Heilongjiang province in 2011.

METHODSA total of 356 rodents were captured by night trap on the Heixiazi island from April to October 2011. The kidney tissue samples were collected by asepsis operation and the genomic DNA were extracted from them. Leptospira strains were confirmed by polymerase chain reaction(PCR) amplification of the 482 bp 23 S rDNA gene. Fifteen PCR products selected by the month were purified and sequenced by the methods of Sanger dideoxy, the sequences then compared with other Leptospira strains in Genebank, and phylogenetic analyses were drafted by software Mega 4.0.

RESULTSAmong 356 rodents, the dominant species were Clethrionomys rutilus (39.3%, 140/356) and Apodemus agrarius (36.0%, 128/356). The infection rate of Leptospira was 11.0%, with 39 rodent samples detected positive. All the rodent species were infected except for Rattus norvegicus. The infection rate was 9.4% (12/128) in Apodemus agrarius, 12.9%(18/140) in Clethrionomys rutilus, 10.8%(7/65) in Microtus fortis Buchner. No significant difference was found between the infection rate and the species of rodents by chi square test(χ(2) = 1.92, P > 0.05). Among months, the infection rate was 5.6% (4/72) in May, 8.8% (5/57) in June, 12.8% (5/39) in July, 9.8% (5/51) in August, 33.3% (11/33) in September, 22.5% (9/40) in October,but no infection in April. There was significant difference in infection in different months (χ(2) = 32.92, P < 0.05). All the Leptospira in rodents on the Heixiazi island were in the same phylogenetic branch with a high similarity of 97.1%-99.6%, close with the Australia strain U90865 by the similarity above 96.3%.

CONCLUSIONLeptospira is probably prevalent in rodents on the Heixiazi island, and the phylogene of the strains were similar. The infection rate in rodents was significantly different in months but not in hosts.

Animals ; China ; Leptospira ; isolation & purification ; Leptospirosis ; prevention & control ; Murinae ; microbiology ; Phylogeny ; Rats

OBJECTIVETo study the clinicopathologic and prognostic significance of serum levels of six cytokines (IFN-γ, TNF-α, IL-10, IL-5, IL-4, IL-2) in patients with advanced serous ovarian cancer prior to surgery.

METHODSThe serum levels of six cytokines were detected in 51 patients with advanced serous ovarian cancer and 46 healthy controls, using cytometric bead arrays.

RESULTSThe serum levels of IFN-γ (20.68±11.45), IL-2 (4.54±1.18), IL-4 (5.66±2.25), IL-5 (2.72±0.86) µg/L and IL-10 (5.93±7.92) µg/L were higher (P<0.01, P<0.05) and the serum level of TNF-α (7.53±8.47) was lower (P<0.01) in patients with advanced serous ovarian cancer than those in the healthy controls. The IFN-γ/IL-4 ratio (3.93±2.34) of the patients was lower than that of the controls (P<0.01). Kaplan-Meier analysis revealed that patient's age (P=0.016), menopausal status (P=0.001) and serum IL-10 level (P=0.010) correlated significantly with patient's survival. Cox regression analysis showed that serum IL-2 (P=0.045) and IL-10 levels (P=0.007) were the independent prognostic factors.

CONCLUSIONSPatients with advanced serous ovarian cancer have Th1/Th2 imbalance and immune function disturbance. The age of patients and menopausal status are important prognostic factors. IL-2 and IL-10 level are also independent predictors of survival.

Adult ; Age Factors ; Aged ; Cystadenocarcinoma, Serous ; blood ; pathology ; surgery ; Cytokines ; blood ; Female ; Follow-Up Studies ; Humans ; Interferon-gamma ; blood ; Interleukin-10 ; blood ; Interleukin-2 ; blood ; Interleukin-4 ; blood ; Interleukin-5 ; blood ; Kaplan-Meier Estimate ; Lymphatic Metastasis ; Menopause ; Middle Aged ; Neoplasm Staging ; Ovarian Neoplasms ; blood ; pathology ; surgery ; Preoperative Period ; Prognosis ; Proportional Hazards Models ; Survival Rate ; Tumor Necrosis Factor-alpha ; blood

CD4-Positive T-Lymphocytes ; Child ; Cryptococcosis ; immunology ; Humans ; Lymphopenia ; etiology

Adolescent ; Adult ; Asian Continental Ancestry Group ; genetics ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Male ; Membrane Transport Proteins ; genetics ; Middle Aged ; Mutation ; genetics ; Polymerase Chain Reaction ; Sequence Analysis, DNA ; Spastic Paraplegia, Hereditary ; genetics ; Young Adult

OBJECTIVETo observe the effects of ginsenoside Rg1 on the functional expression of human neural stem cells (hNSCs).

METHODThe membrane electrophysiological properties and sodium and potassium ion channels in the hNSCs induced by Rg1 were analyzed using the whole-cell patch-clamp.

RESULTOn the 7th day, the neuron-like cells derived from ginsenoside Rg1 (20 mg x L(-1))-induced NSCs show: (1) The resting membrane potential: (-45.70 +/- 2.63) mV, the membrane capacitance: (26.89 +/- 1.91) pF, the membrane input impedance: (877.51 +/- 20.44) MH (P < 0.05 compared with the control group, respectively); (2) The detection rate of inward sodium current which is rapidly activated and inactivated in voltage-dependence was 50%, and its average peak value was (711.48 +/- 158.03) pA (P < 0.05 compared with the control group); (3) The outward potassium currents were composed of rapidly activated and inactivated transient outward potassium current and delayed rectifier outward potassium current, and its average peak value was (1 070.42 +/- 177.18) pA (P < 0.05 compared with the control group).

CONCLUSIONGinsenoside Rg1 can promote the functional expression and maturity of hNSCs.

Cells, Cultured ; Gene Expression ; drug effects ; Ginsenosides ; pharmacology ; Humans ; Membrane Potentials ; drug effects ; Neural Stem Cells ; cytology ; drug effects ; Patch-Clamp Techniques ; Plant Extracts ; pharmacology ; Potassium Channels ; genetics ; metabolism ; Sodium Channels ; genetics ; metabolism

Objective To establish an isotope dilution gas chromatography-mass spectrometry(GC-MS)for the determination of chloropropanol esters in fried foods.Methods A total of 88 fried food samples were collected from supermarket,breakfast shop and street breakfast,stalls,the fried food sample with no chloropropanols esters detected was used as the blank sample. Samples were extracted using a solvent extraction method,followed by ester-bond cleavage reaction with sodium methylate-methanol and purification by diatomite solid-supported liquid-liquid extraction column. The derivatives in purified solution was detected by GC-MS after being derivatived with heptafluoro butyrylimidazole. The concentration of chloropropanols esters was quantified by using deuterium isotopes as internal standards. The accuracy of the method for evaluating recovery rate of blank samples was adopted,and the relative standard deviation(RSD)of the recovery rate represents the precision of the method.Results The 3-MCPD ester and 2-MCPD ester had good linear relationship in the concentration range of 25-1000 g/L(r>0.9995). The detection limits of 3-MCPD ester and 2-MCPD ester were 20μg/kg. The recovery rate of fat extracts from blank samples at 25,50,100,and 200μg/kg levels ranged from 89.7% to 103.7%,and RSD<8.4%. The detection rates of 3-MCPD ester and 2-MCPD ester in 88 samples were 81.82% and 70.45% respectively,and the content ranges from not-detected(ND) to 1.65mg/kg and to 0.93 mg/kg respectively.Conclusion The method is simple,accurate and reliable. It is suitable for the determination of chloropropanol esters in fried foods. There is a certain degree of contamination of chloropropanol esters in fried foods,and this comtamination is quite common.