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.

Using 16S rDNA clone library method, the bacteria composition of the Organic Matter-decomposing Inoculants A and B were investigated. The results indicated that: Sample A was clustered into 14 taxonomic operational units (OTUs),the dominant communities are Weissella confuse, Bacillus subtilis and Bacillus pumilus, which accounted for 28.6%,30.4% and 23.2%;Sample B was clustered into 43 OTUs,the dominant communities are Lactobacillus buchneri, Lactobacillus farciminis and Lactobacillus acetotolerans, which accounted for 18.03%,18.86% and 13.12%, respectively. The results had much difference from the samples' labels:there were not bacteria in the lable of sample A and only Bacillus pumilus was the same with the lable of sample B.This study showed that 16S rDNA clone library method has nicer application perspectives in analying the bacteria of microbial inoculant and its quality control.

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..

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

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.

Oral mucosal drug delivery has the advantages of rapid drug absorption, no first-pass effect and good patient compliance. However, factors such as low drug dissolution, saliva carrying the drug into the gastrointestinal tract and the existence of physiological barriers in the mucosa may affect the mucosal permeation and bioavailability of the drug. Nanotechnology applied to drug oral mucosa delivery can overcome the above disadvantages and obtain efficient absorption effect. This paper describes the physiological structure of oral mucosa and the factors affecting the absorption of drugs in oral mucosa, reviews the application of nanotechnology such as liposomes, solid lipid nanoparticles, nanostructured lipid carriers, nanoemulsions, polymer nanoparticles, polymer micelles and nanohybrid suspensions in oral mucosal drug delivery and the mechanism of promoting drug absorption, summarizes the main problems of current research, and gives an outlook on the application of nano oral mucosal drug delivery system. The main problems of current research are summarized, and the prospects for the application of nano oral mucosal drug delivery systems are discussed.

BACKGROUNDLiver targeting drug delivery systems can improve the curative effects and relieve the cytotoxicity of the chemotherapy drugs in the treatment of liver diseases. Nanoparticles carrying therapeutic drugs are currently under hot investigation with great clinical significance. This study was aimed to investigate the different tissue distribution of the adriamycin polybutylcyanoacrylate nanoparticle (ADM-PBCA-NP) in the mice body after an injection via lateral tail vein, and to study the liver targeting effects of ADM-PBCA-NP in different diameters on normal mice liver.

METHODSOne hundred and eighty Kunming mice were randomly divided into 6 groups with 30 mice in each group (5 treatment groups of ADM-PBCA-NP in the different diameter ranges, non-conjugated free adriamycin injection was employed as the control group). A single dose of either conjugated or free adriamycin equaled 2 mg/kg of body weight was delivered via the tail vein. Five mice in each trail were sacrificed at 5, 15, 30 minutes, 1, 5 and 12 hours postinjection, respectively. The adriamycin concentrations in the respectively collected liver, kidney, spleen, heart, lung and plasma were demonstrated using a high performance liquid chromatography with fluorescence detector.

RESULTSCompared with the control group, adriamycin was hardly detected in the heart muscle of the treatment groups (P < 0.05). The nanoparticle-conjugated adriamycin was cleaned up quickly from the kidney tissue. The adriamycin concentrations of the mice liver and spleen in the experimental groups were significantly higher than that in the control group, except for the group with the nanoparticles diameters of (22.3 +/- 6.2) nm (P < 0.05). The ADM-PBCA-NP in (101.0 +/- 20.3) nm diameter had the highest liver distribution, and the second highest adriamycin distribution in liver was the group of (143.0 +/- 23.5) nm diameter (P < 0.05). Moreover, adriamycin was released slowly in the liver during the detection period in the experimental groups. ADM-PBCA-NP in (22.3 +/- 6.2) nm diameter was not distributed in the tissue of the liver, kidney, heart, spleen, and lung.

CONCLUSIONSADM-PBCA-NP in 100 - 150 nm diameter range has the best liver targeting with a characteristic of slow medicine release. It also decreases the medicine distribution in the heart, kidney and lung. In the treatment of liver cancer, the polybutylcyanoacrylate nanoparticles system has a good liver targeting ability, which increases the anticancer activity and markedly decreases the toxicity of adriamycin.

Animals ; Antineoplastic Agents ; administration & dosage ; Doxorubicin ; administration & dosage ; Drug Delivery Systems ; Enbucrilate ; administration & dosage ; Liver ; metabolism ; Mice ; Nanostructures ; Tissue Distribution

According to the reported sequence of Buthus martensii Karsch scorpion toxin gene (BmK IT3), we synthesized two primers, which were complementary in a region. By the means of PCR, we got the gene. The gene was fused in expression vector pET-28a, which gave rise to a recombinant plasmid pET(IT3R). Then it was transformed into E. coli BL21 (DE3). With IPTG induction, the gene was efficiently expressed. And the fusion product was soluble.
Animals ; Cloning, Molecular ; Escherichia coli ; genetics ; Gene Expression ; drug effects ; Isopropyl Thiogalactoside ; pharmacology ; Recombinant Proteins ; biosynthesis ; genetics ; Scorpion Venoms ; biosynthesis ; chemistry ; genetics ; Scorpions ; chemistry ; genetics

OBJECTIVETo construct the localization system involving anti-TfR monoclonal antibody (McAb) and AFP promoters and assess its effect on human hepatoma cell lines.

METHODSThe conjugate of anti-TfR McAb and polylysine (PLL) was made by SPDP and purified by molecular screen chromatography. DNA blocking test determined that the ratio of one pEBAF/tk to six Ab-PLL was the most suitable to couple them. The pEBAF/tk recombinant plasmid bearing HSV-TK gene was coupled to Ab-PLL by noncovalent bond. The pEBAF/tk was transferred into human hepatoma cell line HepG2, SMMC7721 and pulmonary cancer cell line A549 by receptor-mediated gene delivery (Ab-PLL-DNA) and liposome procedure. The growth inhibitory rates of HepG2, SMMC7721 and A549 cells were measured by MTT assay.

RESULTSThe inhibitory rates of HepG2/tk in 100 mg/L and 1 mg/L of GCV were 60.5% and 24.3%, respectively. The inhibitory rate of GCV to SMMC7721 was 23.2% in 3 days. The pulmonary cancer cell A549, A549/tk (Ab) and A549 /tk (lipo) could not be inhibited by the addition of GCV.

CONCLUSIONThe localization system employed in this paper has high specificity, effectiveness and safety for gene therapy. It would be a promising strategy for gene therapy.

Antibodies, Monoclonal ; therapeutic use ; Carcinoma, Hepatocellular ; therapy ; Cell Line, Tumor ; Ganciclovir ; therapeutic use ; Genetic Therapy ; Humans ; Liver Neoplasms ; therapy ; Receptors, Transferrin ; immunology ; Simplexvirus ; enzymology ; Thymidine Kinase ; genetics ; alpha-Fetoproteins ; genetics

OBJECTIVETo investigate the protective effect and mechanism of Shenqi Compound on diabetic angiopathy modeled rats.

METHODSTotally 18 SD rats were randomized into 3 groups, i.e., the normal control group, the diabetic mellitus (DM) group, and Shenqi Compound group, 6 in each group. The DM rat model was established by feeding high-fat diet (to induce hyperlipidemia) +intraperitoneal injection of small dose streptozotocin (STZ). Shenqi Compound was given to rats in the Shenqi Compound group at the daily dose of 2 g/kg. Equal volume of normal saline was given to rats in the model group and the normal control group by gastrogavage. All treatment was lasted for 12 weeks. Then 2-D and ultrasonic integrated backscatter technique were used to evaluate structural and functional changes of abdominal aorta in the progression of diabetic macroangiopathy. The fibrosis degree of the aorta vessel and myocardium capillaries were observed by using HE and Masson trichrome staining. The tension of the aortic vascular ring was determined. The transforming growth factor beta (TGF-beta) mRNA expression was detected by real time PCR (RT-PCR). The protein expression of TGF-beta, collagen I, collagen III, connective tissue growth factor (CTGF), and phosphorylation P38 MAPK were detected by Western blot.

RESULTSCompared with the normal control group, abdominal aortic systolic inner diameter, diastolic inner diameter, Peterson elastic modulus, stiffness index, and backscatter integral significantly increased; the rangeability of integral backscatter and the extension coefficient of cross section significantly decreased in the DM group (all P < 0.05). After 12 weeks aforesaid indices were obviously improved in the Shenqi Compound group (P < 0.05). Results of HE and Masson staining showed that the fibrosis degree of the aorta vessel and myocardium capillaries was obviously alleviated in rats of the Shenqi Compound group (P < 0.05). Results of the aortic vascular ring tension showed that acetylcholine induced vasodilatation and maximum diastolic percent were obviously elevated in the Shenqi Compound group (P < 0.05). Compared with the normal control group, the mRNA expression of TGF-beta, and the protein expression of TGF-beta, collagen I, and collagen III, and phosphorylation of P38 MAPK all significantly increased in the DM group (P < 0.05). Compared with the DM group, the mRNA expression of TGF-beta, and the protein expression of TGF-beta, collagen I, and collagen III, and phosphorylation of P38 MAPK all decreased (P < 0.05).

CONCLUSIONSShenqi Compound could effectively improve the arterial function in diabetic marcoangiopathy and microvascular dysfunction. The mechanism might be due to the down-regulating the expression of TGF-beta, and further suppressing the phosphorylation of P38 MAPK, reducing the synthesis of collagen I and collagen III, therefore, ameliorating arterial and myocardial interstitial fibrosis.

Animals ; Collagen Type I ; metabolism ; Collagen Type III ; metabolism ; Diabetes Mellitus, Experimental ; drug therapy ; Diabetic Angiopathies ; prevention & control ; Drugs, Chinese Herbal ; pharmacology ; Male ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Transforming Growth Factor beta1 ; metabolism ; p38 Mitogen-Activated Protein Kinases ; metabolism