Objective To study the way of quick identification of haemophilus influenzae with reverse dot blot.Methods Oligonucleotide probe which is specially targeted to 16SrDNA of haemophilus influenzae was designed, then fixed the probe to nylon membranes, and hybridized with the production of gain with the universal primers.Results The universal primers could hybridize the target sequence from common pathogenic bacteria by PCR, and oligonucleotide probe could hybridize with haemophilus influenza specially and could not hybridize with other bacterias. It proved that the probe was of highly speciality.Conclusion Reverse dot blot is a good method of quickly identification of haemophilus influenzae.

Objective To know the expression of 47 kD protein of orientia tsutsugamushi(Ot) Karp strain and test its immunogenicity and antigenicity.Methods Forty-seven kD protein gene that code the whole mat peptide was amplified from the genomic DNA of Ot.Karp strain by PCR and constructed to a recombinant plasmid pGEX-47.The E. coli cells BL21 were transformed with pGEX-47,then the transformants were induced to express the recombinant 47 kD protein by IPTG. The antigenicity and immunogeni-(city) were identified by Dot Blot method and the experiment of immunizing mice respectively.Results 1. The PCR product of 47 kD protein gene of Ot.Karp strain was obtained and its sequence was the same as the published 47 kD protein gene.2. The E.coli expression plasmid pGEX-47 constructed was identified containing the 47 kD protein gene fragment by restrication analysis and PCR identification,and an expression band about 47 kD was detected by SDS-PAGE of the purified product from GST-fusson protein expressed by pGEX-47. 3.Dot Blot analysis and the experiment of immunizing mice testified its antigenicity and immunogenicity.Conclusions The 47 kD protein gene of Ot Karp strain is amplified that code the whole mat peptide, which can be expressed in prokaryotic cells and the expressed protein has antigenicity and immunogenicity.

120 strains of endophytic bacteria identified by ERIC-PCR were isolated from wild and cultivated Glycyrrhiza uralensis plants which collected from Erdos Innermongolia province.The identified results indicated that Glycyrrhiza uralensis plants has plenty of endophytic bacterium in density and population,and the density is higher in root and leave than in stem.Partial sequence analysis of 16S rDNA gene of 82 strains indicated that these strains were in a high similarity with 19 known genus which belong to?、?、 ?-Proteobacteria、Firmicutes and Actinobacteria.The dominant genus were Bacillus sp.,Pseudomonas sp., Pantoea sp.and Serratia sp..

Objective To evaluate the theraputic effect of transcatheter arterial chemoembolization (TACE)for hepatocellular carcinoma with tumor thrombosis of portal vein.Methods One hundred and six patients of hepatocellular carcinoma with tumor thrombosis of portal vein under treament of TACE were observed before and after the procedure.Results After TACE tumor size reduced＞50% in 23 patients,＜50% in 25, no significant change in 44.The size of tumor enlarged in 12.The disappearance of portal vein tumor thrombosis accessed in 14,with reduction in 39,and no significant change in 51.Two patients died within one week.Conclusion TACE provides good therapeutic effect on hepatocellular carcinoma with tumor thrombosis of portal vein.

OBJECTIVEThe rapid identification of pathogenic bacteria is important for earlier effective patient management and antimicrobial therapy, especially for the infant patient, whose immunological system is not fully developed. However conventional microbiogical techniques of bacterial identification, culture and isolation of pathogenic bacteria, identification by biochemistry and serological assay, are time-consuming and require intensive labor. On the basis of special gene sequence, PCR provides simple and rapid way to identify bacteria. But it is difficult to identify all of bacteria species which are suspicious of pathogenic agents. Oligonucleotide arrays provide a powerful tool for parallel detection of target genes. The objective of this study was to test a reverse oligonucleotide assay, which hybridize with the PCR product of 16SrDNA using a pair of universal primers, to rapidly identify common infant pathogenic bacteria.

METHODSBy comparison and analysis of the 16SrDNA sequences of common pathogenic bacteria, a region, which has numerous sequence variations and flanked by highly conserved sequences, was found. A pair of universal primers was designed according to its flanking conservative sequence, and a set of probes specially targeting to eight species of infant pathogenic bacteria, including staphylococcus aureus, Pseudomonas aeruginosa, Klebsiella pneumoniae, Streptococcus faecalis, Hemophilus influenzae, Enterobacter cloacae, Escherichia coli, and Acinetobacter baumannii,according to the variable sequences. The probes were fixed on the nylon membrane with positive electricity, and hybridized them with the products of PCR using the universal primers.

RESULTSThe universal primers could amplify the target sequence from bacteria including the eight common infant pathogenic bacteria and Staphylococcus epidermidis, Enterobacter aerogenes, Streptococcus pneumoniae,beta-hemolytic streptococcus, Neisseria meningitides, Citrobacter freundii, Bacillus subtilis, and Salmonella infantis,but could not amplify rotavirus and human DNA as control. The results showed that the oligonucleotide array could specially hybridize with the eight bacteria to be examined and could not hybridize with other bacteria. The lowest concentration of DNA (product of PCR) for oligonucleotide array was about 25 ng/ml. The results proved that the probes are highly selective and the oligonucleotide arrays could parallelly detect the eight common infant pathogenic bacteria. The results suggested that the oligonucleotide array system was able to identify the eight common infant pathogenic bacteria from clinical specimens and the results were the same as identified by automated bacterial detection machine. From the further experiments, the oligonucleotide array system could directly diagnose the common infant pathogenic bacteria from the broths of samples culture.

CONCLUSIONSDespite limited number of identifiable bacteria and lack of information on antibiotic susceptibility of bacteria, the reverse oligonucleotide assay system, which contains amplification of the segment of 16rDNA from samples using the universal primers and parallel detection of PCR products using specific probes, is an effective method to rapidly identify the eight common infant pathogenic bacteria.

Bacterial Infections ; diagnosis ; microbiology ; Humans ; Infant ; Infant, Newborn ; Oligonucleotide Array Sequence Analysis

Thermosensitive gel with the property of LCST (Lower Critical Solution Temperature) was studied as a hot spot these years for it can be used as one supportor of drug controlled release system which can release drug at right point, right time and right quantity. With further reseaches and applications of thermosensitive gel, the modes and mechanisms of it were discovered and poited out, drug release models were also established step by step. Resesrches on drug release mechanisms and modes of thermosensitive gel will make it applied better in fields of medicine and biology.
Drug Delivery Systems ; methods ; Gels ; Models, Theoretical ; Temperature

The preliminary metabolic profile of total diterpene acid (TDA) isolated from Pseudolarix kaempferi was investigated by using in vivo and in vitro tests. Pseudolaric acid C2 (PC2) was identified as the predominant metabolite in plasma, urine, bile and feces after both oral and intravenous administrations to rats using HPLC-UV and HPLC-ESI/MS(n), and demethoxydeacetoxypseudolaric acid B (DDPB), a metabolite proposed to be the glucoside of PC2 (PC2G), as well as pseudolaric acid C (PC), pseudolaric acid A (PA), pseudolaric acid A O-beta-D glucopyranoside (PAG), pseudolaric acid B O-beta-D glucopyranoside (PBG) and deacetylpseudolaric acid A (DPA) originated from TDA could also be detected. It was demonstrated by tests that the metabolism of TDA is independent of intestinal microflora, and neither of pepsin and trypsin is in charge of metabolism of TDA, TDA is also stable in both pH environments of gastric tract and intestinal tract. The metabolites of TDA in whole blood in vitro incubation were found to be PC2, DDPB and PC2G, which demonstrated that the metabolic reaction of TDA in vivo is mainly occurred in blood and contributed to be the hydrolysis of plasma esterase to ester bond, as well as the glucosylation reaction. These results clarified the metabolic pathway of TDA for the first time, which is of great significance to the in vivo active form and acting mechanism research of P. kaempferi.
Animals ; Chromatography, High Pressure Liquid ; Diterpenes ; metabolism ; Glucosides ; metabolism ; Hydrolysis ; Mass Spectrometry ; Metabolic Networks and Pathways ; Pinaceae ; chemistry ; Rats

98 endophytic bacteria strains were isolated from different internal tissues of Glycyrrhiza uralensis plants collected from Innermongolia region. Results indicated that the population densities of endophytic bacteria ranged from 5.0?104cfu/g～2.9?107cfu/g fresh weight although it varied depending on tissue of the plant. Among these strains, Bacillus sp. was the most prevalent endophytic bacterium, which was amount to 30%.Of the 98 isolates, 6 strains exhibited extensive antagonistic activities against pathogenic bacteria. Characterization showed that these bacteria were Bacillus atrophaeus、Paenibacillus polymyxa、Bacillus subtilis、Paenibacillus ehimensis. This study indicated that selected 6 endophytic bacteria strains have potential for biological control of plant disease.

Objective To study the expression and the relation of vascular endothelial growth factor (VEGF)and matrix metal proteinase-2(MMP-2)in rectal cancer and evaluate their roles in rectal carcinogen- esis and development.Methods The expression of VEGF and MMP-2 in 52 cases of rectal cancer was de- tected by immunohistochemical SP technique.12 cases normal rectal tissue served as the control group.Re- suits The expression of VEGF in rectal carcinoma(67.3 %)was much higher than that in control group(P

OBJECTIVETo investigate the effects of beta1-integrin, fibronectin (FN) and laminin (LN) on the invasive behavior of human gliomas.

METHODSFunctional impacts of beta1-integrin, fibronectin and laminin on cell adhesion, migration and metastasis of U251 malignant glioblastoma cells were investigated by in vitro adhesion, migration and invasion assays. The amount and distributions of cellular microfilaments and pseudopodia were studied by fluorescent cytochemistry, confocal laser scanning microscope and scanning electron microscope. Lastly, beta1-integrin, fibronectin and laminin were investigated for their roles in cellular microfilament skeleton.

RESULTS(1) Fibronectin did not affect cell adhesion of U251MG cells, but anti-beta1 integrin antibodies inhibited cell adhesion (P < 0.01); Laminin stimulated cell adhesion of U251MG cells (P < 0.01) but anti-beta1 integrin antibodies had little effect on the laminin-mediated cell adhesion. (2) The migration of U251MG cells on dishes coated with FN was inhibited by anti-beta1 integrin antibodies (P < 0.05). (3) F-actins formed strong and dense stress fibers in U251MG cells on dishes coated with FN and LN. Anti-beta1 integrin antibodies disrupted the microfilament network and F-actin aggregation. (4) FN and LN increased the number of pseudopodia on cell surface, whereas anti-beta1 integrin antibodies reversed this function. (5) FN and anti-beta1 integrin antibodies had little effects on the invasive ability of U251MG cells in vitro. The invasion was increased by LN, but inhibited by anti-beta1 integrin antibodies.

CONCLUSIONS(1) The interaction between beta1-integrin, FN may stimulate U251MG cell migration via changing the structures of microfilament skeleton and the number of pseudopodia. (2) beta1-integrin may play a role in the LN-mediated in vitro invasion of U251MG cells.

Cell Adhesion ; drug effects ; Cell Line, Tumor ; Cell Movement ; drug effects ; Fibronectins ; pharmacology ; Glial Fibrillary Acidic Protein ; analysis ; Glioma ; metabolism ; pathology ; ultrastructure ; Humans ; Immunohistochemistry ; Integrin beta1 ; pharmacology ; Laminin ; pharmacology ; Microscopy, Confocal ; Microscopy, Electron, Scanning ; Neoplasm Invasiveness