[EMA-pCR detection of enterohemorrhagic Eschrichia coli O157:H7].

Objective: A rapid and effective method with ethidium monoazide bromide (EMA) in combination with PCR (EMA-PCR) was established to detect live Enterohemorrhagic Eschrichia Coli O157:H7.

Methods: The rfbE gene was used as the target gene for PCR detection of Eschrichia Coli O157:H7 by utilizing its pure isolates after the treatment of EMA as the template. The EMA concentration and reaction time was optimized.

Construction and secretory expression of beta-galactosidase gene from Lactobacillus bulgaricus in Lactococcus lactis.

Objective: This study is to examine the secretion effects of beta-galactosidase in Lactococcus lactis.

Methods: The usp45 and beta-galactosidase genes were cloned and inserted into plasmid pMG36e to obtain the recombinant plasmid pMG36e-usp-lacZ. This recombinant plasmid was transformed into both Escherichia coli DH5alpha and L.

[Investigate the effects of compound radix notoginseng on renal interstitial fibrosis and kidney-targeting treatment].

Objective: Investigate the effects of compound Radix Notoginseng on renal interstitial fibrosis and kidney-targeting treatment.

Methods: 100 healthy Sprague-Dawley rats were randomly divided into 5 groups: Unilateral ureteral obstruction (UUO) group, sham-operation (SOR) group, Radix Notoginseng (RN) group, compound Radix Notoginseng (CRN) group and Losartan (ARB) group. After operation, RN, CRN and ARB groups were intragastric administrated with RN (3 mL/d), CRN (3 mL/d) and ARB [20 mg/(kg x d)] respectively.

Analysis of β-galactosidase production and their genes of two strains of Lactobacillus bulgaricus.

A bacterial β-galactosidase delivery system is a potential therapy for lactose intolerance. Currently, two Lactobacillus bulgaricus strains with different biological characteristics are under consideration as potential sources. However, differences in these β-galactosidase genes and their resulting production levels are poorly characterized.

[Hemagglutinin gene of influenza A (H3N2) virus from human samples in the 2009 influenza epidemic in Chengdu].

Objective: To study the antigenic and genetic characteristics of influenza A (H3N2) virus in the 2009 influenza epidemics in Chengdu.

Methods: The influenza virus strains were isolated with MDCK cells from 4869 samples taken from the sentinel surveillance in 2009 in Chengdu. Hemagglutination inhibition (HI) and RT-PCR reaction tests were performed to guide the extraction of viral RNA from the culture fluid of the influenza A (H3N2) virus.

Ginsenoside Rg1 modulation on thrombospondin-1 and vascular endothelial growth factor expression in early renal fibrogenesis in unilateral obstruction.

Renal interstitial fibrosis is the major histopathological change seen in a variety of renal disorders and is closely related to renal dysfunction. Progressive interstitial fibrosis accompanied by the loss of renal tubules and interstitial capillaries typifies all progressive renal disease. Thrombospondin-1 (TSP-1) is a major angiogenic inhibitor.

LSKL, a peptide antagonist of thrombospondin-1, attenuates renal interstitial fibrosis in rats with unilateral ureteral obstruction.

The effects of LSKL, the peptide antagonist of thrombospondin-1 (TSP-1), on renal interstitial fibrosis in rats subjected to unilateral ureteral obstruction (UUO) were investigated. Rats were divided randomly into three groups (n = 20 each): UUO group, sham-operation group and UUO plus LSKL treatment group. Collagen deposition was studied using histopathology and reverse transcription polymerase chain reaction analysis (RT-PCR).

[Development of a universal primers PCR-coupled liquid bead array to detect biothreat bacteria].

Objective: To develop a fast, high-throughput screening method with suspension array technique for simultaneous detection of biothreat bacteria.

Methods: 16 S rDNA universal primers for Bacillus anthracis, Francisella tularensis, Yersinia pestis, Brucella spp.and Burkholderia pseudomallei were selected to amplify corresponding regions and the genus-specific or species-specific probes were designed.

[Carrying rate of Neissria meningitides in a healthy population in the Mianzhu post-earthquake residential area].

Objective: To predict the possibility of epidemic outbreak of meningitis by testing Neissria Meningitides in a healthy population in the Mianzhu post-earthquake residential area.

Methods: A simple random sampling strategy was adopted to collect 887 throat swabs from a healthy population in the Mainzhu post-earthquake residential area. The TaqMan assay were performed to detect Neissria Meningitides.

[Development of a multiplex PCR-suspension array for simultaneous detection of five bioterrorism bacteria].

Objective: To develop a rapid, high-throughput screening method of gene suspension array technique to simultaneously detect five bioterrorism bacteria: Bacillus anthracis, Francisella tularensis, Yersinia pestis, Brucella spp. and Burkholderia pseudomallei.

Methods: Highly validated specific primers were used to amplify diagnostic regions unique to each pathogen.

Ginsenoside Rb1, a panoxadiol saponin against oxidative damage and renal interstitial fibrosis in rats with unilateral ureteral obstruction.

Objective: To investigate the possible protective effect and mechanism of ginsenoside Rb1 against oxidative damage and renal interstitial fibrosis on rats with unilateral ureteral obstruction (UUO).

Methods: In total, 80 male rats were randomly divided into 4 groups, 20 in each group: the sham operated group (SOR), UUO group, UUO with ginsenoside Rb1 treatment group (treated with intraperitoneal injection of 50 mg/ kg daily) and UUO with Losartan treatment group (as the positive control, treated with 20 mg/kg by gastrogavage per day). The rats were randomly sacrificed on day 3, 7 and 14 after surgery, respectively.

[Effects of ginsenoside Rb1 on TGF-beta1 induced p47phox expression and extracellular matrix accumulation in rat renal tubular epethelial cells].

Objective: To investigate the effects of Ginsenoside Rb1 (G-Rb1) on the oxidative damage and extracellular matrix accumulation in rat renal tubular epethelial cells induced by transforming growth factor-beta1 (TGF-beta1).

Methods: Cultured normal rat renal tubular epethelial cells (NRK-52E) were divided into control group, 10 ng/mL TGF-beta1-induced group, G-Rb1 treated groups in which rat renal tubular epethelial cells were treated with different concentration of G-Rb1 (10 ng/mL, 20 ng/mL, 40 ng/mL) after TGF-beta1 induction, G-Rb1 40 ng/mL group and 100 nmol/L DPI(diphenyleneiodonium, an inhibitor of NADPH oxidase) group. Intracellular reactive oxidative species (ROS) level was measured by flowcytometry.

[Construction and property study of recombinant Lactococcus lactis with non-fusion expressing of beta-galactosidase].

Objective: To construct recombinant Lactococcus lactis strains exhibiting high beta-galactosidase activity in non-fusion way, and study their enzyme activities and enzyme secretion rates.

Methods: The recombinant plasmids pMG36e-lacZ 1.1480 and pMG36e-lacZ wch9901 which could express beta-galactosidase from Lactobacillus delbrueckii subsp.

Non-fusion and fusion expression of beta-galactosidase from Lactobacillus bulgaricus in Lactococcus lactis.

Objective: To construct four recombinant Lactococcus lactis strains exhibiting high beta-galactosidase activity in fusion or non-fusion ways, and to study the influence factors for their protein expression and secretion.

Methods: The gene fragments encoding beta-galactosidase from two strains of Lactobacillus bulgaricus, wch9901 isolated from yogurt and 1.1480 purchased from the Chinese Academy of Sciences, were amplified and inserted into lactococcal expression vector pMG36e.

Ginsenoside Rg1, a major active component isolated from Panax notoginseng, restrains tubular epithelial to myofibroblast transition in vitro.

The medicinal herb, Panax notoginseng, has been used for thousands of years in traditional Chinese medicine and possesses anti-fibrosis properties. Epithelial-myofibroblast transition (EMT) plays an important role in renal tubulointerstitial fibrosis. The present study was designed to examine whether ginsenoside Rg1, a major active component isolated from Panax notoginseng, has an ability to block this phenotypic transition in rat renal tubular epithelial cells (NRK-52E) induced by transforming growth factor-beta1 (TGF-beta1).

[Ginsenoside R(g1) inhibit transdifferentiation in rat renal tubular epethelial cells induced by TGF-beta1].

Objective: To investigate the effects of ginsenoside R(g1) on the transdifferentiation of rat renal tubular epethelial cells induced by transforming growth factor-beta1, (TGF-beta1).

Method: Cultured normal rat renal tubular epethelial cells (NRK-52E) were divided into control group, TGF-beta1-induced group and treated with ginsenoside R(g1) at different concentration (10, 20, 40 mg x L(-1)) group. The morphology of tubular epithelial-myofibroblast transdifferentiation induced by TGF-beta1 was observed through light microscope.

[Development of a molecular beacon real-time PCR for the rapid detection of Mycobacterium tuberculosis].

Objective: To develop a molecular beacon real-time PCR for rapid detection of Mycobacterium tuberculosis.

Method: One set of primers was selected from the IS6110 gene in GenBank and the corresponding molecular beacon probe was designed. The specificity and sensitivity of the developed method were evaluated by tested with 10 different bacteria species.

[Beta-galactosidase gene from Lactobacillus delbrueckii subsp. bulgaricus gets non-fusion expression in Escherichia coli].

Objective: To construct the food-grade recombinant probiotic strain with high activity beta-galactosidase, the beta-galactosidase gene (lacZ)from Lactobacillus delbrueckii subsp. bulgaricus was in non-fusion expressed in Escherichia coli.

Methods: From Lb.

[Phylogeny analysis and identification of two bacterial strains sourcing from human intestine and having resistance to acid and bile].

Objective: To screen bacteria for the engineering bacteria expressing and secreting high activity beta-galactosidase, and two bacterial strains called as R92-2 and R111 with acid and bile resistances would be isolated from health human intestine to strain identification and phylogenetic analysis.

Methods: These two strains were first been identified with phenotype characteristic analysis. Then the 16S rDNAs of these two bacterial strains were amplified and sequenced with the primers designed by the conserve sequences.

[The effect of ginsenoside Rg1 on the renal interstitial fibrosis of UUO rat].

Objective: To study the effect of ginsenoside Rg1 on the renal interstitial fibrosis caused by unilateral ureteral obstruction (UUO) of rats.

Methods: 80 healthy Sprague-Dawley rats were randomly divided into 4 groups: the UUO group (UUO), sham-operation group (SOR), ginsenoside Rg1 group (Rg1) and losartan group (ARB). From the first day after initial UUO, ARB group was intragastrically administrated with losartan 20 mg/(kg x d).

[Antibiotic resistance and plasmid profiles of Escherichia coli in healthy human bodies].

Objective: To test the antibiotic resistance of 150 Escherichia coli strains isolated from the faeces of healthy people in Shenzhen city.

Methods: Escherichia coli were isolated and identified through a procedure described in the Microbiology in Preventive Medicine and Inspection Technique. The antimicrobial susceptibility of 150 isolates to 16 antibiotics was determined by the Kirby-Bauer method based on the Performance Standards for Anti-microbial Susceptibility Testing recommended by NCCLS.

[Factors influencing the use of health care services from patients with COPD in Chengdu].

Objective: To investigate the use of health care services by COPD patients and to identify factors that shape the pattern of services utilization.

Methods: All of the COPD patients who lived in the six randomly selected communities with community health services in 2003 were surveyed. A total of 446 patients completed the interview-administered questionnaires.

[Effect of Psidium guajava leaf extract on alpha-glucosidase activity in small intestine of diabetic mouse].

Objective: To investigate the inhibition effect of Psidium guajava linn (PGL), a leaf water-soluble extract, on the activities of alpha-glucosidases.

Methods: The PGL water-soluble extract (PGL WE) was obtained by the procedure of distilled water immersion, filtration, extracted fluid concentration and dry of Psidium guajava leaf. The diabetes of Kunming mice was induced by intraperitoneal injection of Streptozotocin (STZ).

[Antimicrobial resistance and molecular epidemiology of coagulase-negative Staphylococci in Chengdu].

Objective: To investigate the antimicrobial resistance and molecular prevalence of coagulase-negative Staphylococcus (CONS) in Chengdu.

Methods: Disk diffusion method was used to test antimicrobial resistance of 2038 CoNS isolated from 15 hospitals and 6 CDCs (center for disease control and prevention) in Chengdu, the results were interpreted according to the guideline of National Committee for Clinical Laboratory Standards. CoNS were typed with plasmid and PFGE (pulsed field gel electrophoresis) profiles.

[Study on the resistance of Escherichia coli O157:H7 to disinfectants in relationship with its plasmid pO157 and chromosome DNA].

Objective: To study the change of Escherichia coli O157:H7 resistance to disinfectants in continuous disinfection and the relationship between the resistance and pO157, chromosome DNA.

Methods: Using 4 disinfectants in a separate manner to disinfect 5 Escherichia coli O157:H7 50 generations continuously, we made a before-after comparison of their resistibility and analyzed the change of the structure of pO157 and chromosome DNA.

Results: After the 50-generation-continuous disinfection, the bacteria resistance to sodium dichloroisocyanurate, iodophor and quaternary ammonium increased, but the resistance to chlorhexidiniacetas did not any change; the maps Cal I and Rsr II cutting pO157 revealed some changed after disinfection by sodium dichloroisocyanurate and quaternary ammonium, but the maps showed no change after disinfection by iodophor and chlorhexidiniacetas; the chromosome DNA PFGE maps change considerably after 50 generations of disinfection by sodium dichloroisocyanurate, iodophor and quaternary ammonium, but the chromosome DNA PFGE maps were similar after 50 generation of disinfection by chlorhexidiniacetas.