The dynamic of endophytic bacteria at different growth stage of tomato and use of these endophytic bacteria to control tomato bacterial wilt were studied. The results showed that endophytic bacteria could be found in the tomato seeds and their quantities reached the highest peak in the adult plants both in resistant and susceptible cultivars. The amount of endophytic bacteria in adult plants of resistant tomato cultivars was 2.43?10~5CFU/g FW in the root and 22.9?10~4 CFU/g FW in the stem, while the amount of endophytic bacteria in adult plants of susceptible tomato cultivars was 9.8?10~4CFU/g FW in the root and 13.4?10~4CFU/g FW in the stem respectively. Seventeen strains of endophytic bacteria from resistant cultivars and only seven strains from susceptible cultivars were found to be antagonistic to Ralstonia solanacearum. In addition, some strains of endophytic bacteria had the abilities of promoting tomato seed germination and controlling tomato bacterial wilt, among which, strain 5R and 3R had better control effect of 91.7% and 81.3% respectively.

Objective To establish a method of detecting hepatitis B virus x antigen (HBxAg) and antibody to HBxAg (anti-HBx) and to demonstrate its clinical significance of HBxAg and anti-HBx in sera from patients of chronic hepatitis B (CHB),liver cirrhosis (LC) and hepatocellular carcinoma (HCC). Methods Full length HBx gene was cloned into pET30a(+),a prokaryotic expression vector,named pET30a-X.It was transformed into Escherichia coli BL21 (DE3),followed the fusion protein of HBx-His was induced by IPTG.The purified fusion protein was used to immunize rabbit as an antigen to generate polyclonal antibody to HBx protein.The method of enzyme-linked immunosorbent assay (ELISA) was established by using purified fusion protein and generated antibody,which was used to detect HBxAg and anti-HBx in sera from patients of CHB,LC,HCC and normal healthy people.Results The positive rates of HBxAg/anti-HBx were 8.7%/10.4% for CHB,17.9%/40.6% for LC,and 9.8%/34.4% for HCC, respectively.In statistics,the positive rates of anti-HBx in LC and HCC were higher than that in CHB (P

OBJECTIVETo evaluate the effects of the Chinese herbal formula Wuzi Yanzong Pill (, WYP) on the spermatogenesis and specific secretory functions of Sertoli cells in rat model and to investigate the underlying mechanism.

METHODSFive groups of male Sprague-Dawley rats including the control group, the model group, the low-dose WYP group, the medium-dose WYP group and the high-dose WYP group (5 in each group) were treated daily with vehicle, multiglycosides of Tripterygium wilfordii Hook f (GTW) either alone (20 mg/kg) or followed by WYP (0.5, 1.0, or 2.0 g/kg daily), respectively for 30 days. Serum levels of follicle-stimulating hormone (FSH), inhibin B (INHB) and testosterone (T) were evaluated using enzyme-linked immunosorbent assay. Androgen-binding protein (ABP) gene expression and transferrin (TF) protein expression in testis tissue specimens of all rats were determined using real-time reverse transcriptase polymerase chain reaction and Western blotting analysis, respectively. Histopathological alterations in the testis were determined using Johnsen's score.

RESULTSThe toxicity of GTW towards Sertoli cell secretory functions and spermatogenesis was accompanied by increased serum FSH concentrations and decreased INHB and T concentrations. Upregulated ABP mRNA levels, and decreased TF protein expression and Johnsen's scores were detected in the model group compared with the control group P<0.05 or P<0.01). Oral high-dose WYP administrations to GTW-treated rats effectively alleviated all of the GTW-induced changes in specific secretory functions of Sertoli cells (ABP, INHB and TF). Furthermore, serum T level and Johnsen's score of the testis increased greatly compared with the model group (P<0.01).

CONCLUSIONWYP has the ability to improve the spermatogenesis, possibly through modulating the secretory proteins expression of Sertoli cells.

Androgen-Binding Protein ; genetics ; metabolism ; Animals ; Blotting, Western ; Drugs, Chinese Herbal ; pharmacology ; Follicle Stimulating Hormone ; blood ; Gene Expression Regulation ; drug effects ; Inhibins ; blood ; Male ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Sertoli Cells ; drug effects ; secretion ; Spermatogenesis ; drug effects ; Tablets ; Testis ; cytology ; metabolism ; Testosterone ; blood ; Transferrin ; metabolism

AIMTo study the excretion of (-)-clausenamide in rats.

METHODSThe urine, feces and bile were collected at predetermined time points after (-)-clausenamide was orally administrated to 6 rats (30 mg x kg(-1)). The concentrations of (-)-clausenamide and its metabolite 6-OH-(-)-clausnamide were determined by HPLC-MS/MS method using glipzide as the internal reference, and the accumulative excretion amount of (-)-clausenamide and 6-OH-(-)-clausenamide was calculated in the urine, feces and bile, separately.

RESULTS(-)-Clausenamide was recovered mostly (44%) from feces in 112 hours, 7.1% was found from urine in 120 hours and 0.013% was detected from bile in 24 hours. The accumulative excretions of 6-OH-(-)-clausenamide were 0.92% , 0.46% and 0.0003% of the administered dose from feces, urine and bile, respectively.

CONCLUSIONThe major amount of (-)-clausenamide was recovered from feces after (-)-clausenamide was orally administrated to rats (30 mg kg(-1)).

Administration, Oral ; Animals ; Bile ; metabolism ; Chromatography, High Pressure Liquid ; Clausena ; chemistry ; Feces ; chemistry ; Female ; Lactams ; chemistry ; pharmacokinetics ; urine ; Lignans ; chemistry ; pharmacokinetics ; urine ; Male ; Mass Spectrometry ; Neuroprotective Agents ; administration & dosage ; pharmacokinetics ; urine ; Plant Leaves ; chemistry ; Plants, Medicinal ; chemistry ; Rats ; Rats, Sprague-Dawley ; Stereoisomerism

AIMTo study the four diastereomers of leucogen and structure of the related substances.

METHODSLC-DAD, LC-MS/MS and LC- 1H NMR were used. LC was carried out with a Phenomnex Luna C18 (250 mm x 4.60 mm ID, 5 microm) column and a mobile phase of water-acetonitrile-glacial acetic acid (58:42: 0.3).

RESULTSThe structures of leucogen and its related substances were identified. Leucogen and the related substances were found to have four diastereomers in solution state separately. The stability and transformation of the four diastereomers were analyzed and 3R4S5R was found to be more stable than the others according to quantum calculations.

CONCLUSIONLeucogen have four diastereomers in solution state and it can transform from one diastereomer to the others, and the 3R4S5R is more stable than the others.

Chromatography, Liquid ; Magnetic Resonance Spectroscopy ; Molecular Structure ; Quantum Theory ; Solutions ; Spectrometry, Mass, Electrospray Ionization ; Stereoisomerism ; Thiazolidines ; analysis ; chemistry

OBJECTIVETo study the loss of heterozygosity (LOH) on chromosome 3p in thyroid tumors.

METHODSLOH at 11 microsatellite loci was analyzed in 74 cases of thyroid tumors (including 20 follicular adenomas, 24 follicular thyroid carcinomas and 30 papillary thyroid carcinomas) by polymerase chain reaction and silver stain.

RESULTSLOH on chromosome 3p was detected in 71% of follicular thyroid carcinoma (17/24), 30% of the papillary thyroid carcinoma (9/30) and 10% of the follicular adenoma (2/20) case. Two minimal common deleted regions (CDR) (3p26-pter and 3p14.2-3p22) involving significant sites of LOH has identified in follicular thyroid carcinoma. There was also one CDR (3p25. 2-26.1) in papillary thyroid carcinoma.

CONCLUSIONSLOH is more frequently identified in follicular thyroid carcinoma than in papillary thyroid carcinoma and follicular adenoma. The 3 CDR on chromosome 3p may harbor tumor suppressor genes involved in the pathogenesis of follicular thyroid carcinoma and papillary thyroid carcinoma.

Adenocarcinoma, Follicular ; genetics ; Adenoma ; genetics ; Adult ; Aged ; Carcinoma, Papillary ; genetics ; Chromosome Mapping ; Chromosomes ; Chromosomes, Human, Pair 3 ; genetics ; Female ; Genes, Tumor Suppressor ; physiology ; Heterozygote ; Humans ; Loss of Heterozygosity ; Male ; Microsatellite Repeats ; Middle Aged ; Thyroid Neoplasms ; genetics ; Young Adult

Objective To investigate existence and homology of 16 S rRNA methylase genes in carbapenem -resistant Klebsiella-pneumoniae ( CRKP ) clinical isolates.Methods Twenty -nine CRKPs were col-lected from four hospitals in Nanchang.PCR amplifications of drug resist-ant genes were performed.The horizontal transmission of the resistance plasmids was evaluated by conjugation and homology of the isolates was analyzed by pulsed field gel electrophoresis ( PFGE).Results Resistant rates of twenty -nine clinical isolates to amikacin and gentamicin were 37.9%( 11/29 ) and 69.0%( 20/29 ) , respectively.The amikacin re-sistanted isolates were simultaneously resistant to gentamicin.The exist-ence of 16 S rRNA methylase positive genes were detected in 10 isolates , including 9 strains carrying armA genes,1 strain carrying rmtB gene.All the 10 strains with 16 S rRNA methylase positive genes harbored β-lac-tamase genes and 7 strains also harbored carbapenemase genes , with blaKPC-2 and blaNDM-1 being the main genotypes.Ten experimental strains were successfully typed by PFGE and classified into 9 different genotypes and resistance plasmids were successfully transferred into the recipient E.coli J53 through conjugation experiments of 3 armA -positive isolates. Conclusion The 16S rRNA methylase gene is highly relevant to carbapenem -resistant klebsiella -pneumoniae in terms of resistance to aminoglycosides , and armA is the main genotype.Plasmids carried 16 S rRNA methylase gene can be horizontally disseminated.

Objective To establish the rapid identification of Acineto-bacter baumannii by detecting specific volatile metabolites of Acinetobacter baumannii through surface desorption atmospheric pressure chemical ioni-zation mass spectrometry ( SDAPCI-MS).Methods Ten clinical isola-ted Acinetobacter baumannii were resuscitated and vaccinated to blood agar plates at 37 ℃incubation and culturing.SDAPCI-MS was adopted to detect volatile metabolites of Acinetobacter baumannii by different cul-turing time.Results Compared with blank culture medium , the pecu-liar volatile metabolites were detected ,the characteristic peaks are mass -to-charge ratio 60 and 118.Basically the same volatile metabolites of Acinetobacter baumannii were generated by different culturing time.Con-clusion The rapid identification of Acinetobacter baumannii through peculiar volatile metabolites is hopefully applied by SDAPCI -MS.

OBJECTIVETo study the effect of co-blockage of vascular endothelial growth factor (VEGF) and its receptor (KDR) on growth of bladder carcinoma T24 cells and nude mice xenograft.

METHODST24 cell line co-transfected with VEGF siRNA and sKDR expression plasmids was developed and its proliferation was assayed by MTT and apoptosis by FCM. The nude mice model bearing bladder carcinoma xenograft was established. The tumor cell VEGF expression, stroma microvessel density (MVD) and tumor cell topoisomerase II alpha (Topo II alpha) expression were detected by immunohistochemistry. Cell apoptosis was estimated by TUNEL assay.

RESULTSMTT assay showed that cell proliferation in VEGF siRNA, sKDR and combination groups was 56.3% +/- 8.3%, 42.6% +/- 13.8% and 32.5% +/- 4.3%, respectively, significantly lower than that in the scramble control (97.3% +/- 11.6%, P < 0.0001). FCM showed there were sub-diploid apoptotic peaks before G1 phase in VEGF siRNA, sKDR and combination groups, and apoptosis ratio was 5.1% +/- 0.9%, 4.2% +/- 0.5% and 8.8% +/- 0.7%, respectively, all of which were higher than that in the scramble control (0.9% +/- 0.4%, P < 0.05), and the combination group had even more higher apoptosis than the two singlely treated groups (P < 0.01). In vivo test showed that tumor growth was inhibited in VEGF siRNA, sKDR and combination groups, and from day 16 the tumor volume in combination group was significantly smaller than that in scramble control (P < 0.05), and from day 28 the tumor almost lost the ability to further growth. Immunohistochemistry revealed VEGF expression in combination group was 54.37 +/- 5.28, significantly lower than that in the scramble control (141.66 +/- 8.59, P < 0.0001). MVD number was only 8.22 +/- 3.79, much less compared with that in the scramble control (61.76 +/- 5.28, P < 0.0001) or sKDR group (19.46 +/- 4.16, P = 0.0089). Tumor cell proliferation index in the combination group (1.5% +/- 0.7%) was significantly decreased compared with that in the scramble control (11.8% +/- 5.2%, P < 0.0001), and apoptosis index (67.2% +/- 8.5%) was much higher than that in the scramble control (8.7% +/- 2.7%, P < 0.0001), VEGF siRNA group (54.3% +/- 4.8%, P = 0.0492) or sKDR group (52.3% +/- 6.4%, P = 0.0293).

CONCLUSIONVEGF siRNA or sKDR alone can inhibit tumor cell proliferation and induce cell apoptosis, but co-blockage of VEGF and KDR by their combination shows more significant therapeutic efficacy.

Animals ; Apoptosis ; Cell Line, Tumor ; Cell Proliferation ; Gene Expression Regulation, Neoplastic ; Humans ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; Neovascularization, Pathologic ; prevention & control ; Plasmids ; RNA Interference ; RNA, Small Interfering ; genetics ; Transfection ; Tumor Burden ; Urinary Bladder Neoplasms ; metabolism ; pathology ; Vascular Endothelial Growth Factor A ; genetics ; metabolism ; Vascular Endothelial Growth Factor Receptor-2 ; genetics ; metabolism ; Xenograft Model Antitumor Assays

OBJECTIVETo test the sensitivity and specificity of alpha-fetoprotein (AFP) colloidal gold diagnostic strip as compared with RIA-AFP diagnostic kit.

METHODSDouble blind method was used to test sera from 426 healthy people and 1567 patients selected in two general hospitals and a cancer hospital. Each serum was measured by RIA-AFP and AFP colloidal gold diagnostic strip on the same day. RIA-AFP diagnostic results was identified as true positive or negative.

RESULTSAmong 426 healthy people, all the RIA-AFP test showed negative result (serum AFP concentration less than 25 IU/ml), but the AFP colloidal gold diagnostic strip had 1.88% false positive. When comparing the result from 1567 patients measured by RIA-AFP, the sensitivity and specificity of AFP colloidal gold diagnostic strip were 99.3%, and 97.2%, respectively. The crude correspondence rate between the two diagnostic regents was 97.6%.

CONCLUSIONAFP colloidal gold diagnostic strip showed very good result and could be used as a screening diagnostic kit in clinic and hospital settings.

Biomarkers, Tumor ; blood ; Carcinoma, Hepatocellular ; diagnosis ; Double-Blind Method ; Gold Colloid ; Humans ; Liver Neoplasms ; diagnosis ; Radioimmunoassay ; Reagent Kits, Diagnostic ; Reproducibility of Results ; Sensitivity and Specificity ; alpha-Fetoproteins ; analysis