OBJECTIVEThe purpose of this research was to study the genetic diversity of F-ATPase subunit gene uncEBF derived from Streptococcus mutans (S. mutans) clinical isolates, furthermore to investigate the relationship between the genetic diversity of F-ATPase and S. mutans aciduric ability.

METHODS38 S. mutans strains included 18 high acid tolerance strains and 20 low acid tolerance strains. Gene uncEBF of these isolates were amplified with specific primers from S. mutans genomic DNA, and the PCR products were analyzed by RFLP and sequenced. SPSS 11.0 statistic software assayed the results.

RESULTSIt was testified that two genotypes A and B of PCR-RFLP were revealed when digested with Alu I and Dde I digested fragments of uncEBF displayed two different patterns C and D. Fisher exact two-tail test showed that the distributions of A and B genotype strains with different acidurance were different (P < 0.05), and the proportion of A genotype strains from high acidurance group was higher than that from low acidurance one. Some of these amplified uncEBF genes from different genotype were sequenced and testified that there existed variation of Alu I and Dde I recognized sites.

CONCLUSIONThis study indicated that uncEBF gene of S. mutans F-ATPase obviously exhibited genetic diversity.

Adenosine Triphosphatases ; Dental Caries ; Genetic Variation ; Genotype ; Humans ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; Streptococcus mutans

OBJECTIVETo study the genetic diversity and the gene expression of membrane-bound proton-translocating ATPase (F-ATPase) subunit gene uncG derived from Streptococcus mutans (S. mutans) clinical isolates.

METHODS38 S. mutans strains derived from caries-active and caries-free individuals including 18 strains displaying high acid tolerance and 20 strains displaying low acid tolerance. Gene uncG was amplified with specific primers from S. mutans genomic DNA, then the PCR product was analyzed by RFLP and sequenced. The relative expression quantity of uncG gene against the housekeeping gene recA was determined by using RT-PCR method. A gel documentation system and QUANTITY ONES software were used to analyze the data results.

RESULTSIt was testified that four genotypes A, B, C and D of PCR-RFLP were revealed when respectively digested with Alu I and Bsr I, but the distributions of the four genotype strains showed no difference (P > 0.05). The differences of uncG gene transcript quantities derived from different genotype or different aciduranc strains had no significance (P > 0.05).

CONCLUSIONThis study indicated that uncG gene of F-ATPase obviously displayed genetic diversity and existed polymorphism at mRNA expression level, while the Alu I-RFLP genotypes and the expression levels would not be responsive to different acid tolerance of S. mutans strains.

Adenosine Triphosphatases ; Dental Caries ; Genetic Variation ; Genotype ; Humans ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Polymorphism, Restriction Fragment Length ; RNA, Messenger ; Streptococcus mutans

OBJECTIVETo analysis the homology among the extended-V region of the surface proteins in different serotype Streptococcus mutans (c, f, d, g) and to find out it's significance in anti-caries vaccine.

METHODSThe DNA of the bacteria (standarded serotype c, d, f, g and partial serotype c clinicals) was extracted and the extended-V region (SrV+, 1 384-2 514 bp) was amplified using polymerase chain reaction (PCR). Then the products were assessed using restriction fragment length polymorphism (RFLP) by endonuclease Dde I. The genotypings were sequenced and analysised using the program of BLAST on NCBI Gene Bank database.

RESULTSAbout 1.13 kb fragments were produced both in serotype c and f, the serotype d and g were failed. The RFLP results showed that five different patterns(A, B, C, D, E) among the 117 PCR products were reveled by Dde I. The ration of the genotypings A and B were the most among the strains, the C was lower, the D and E respectively was 1 and 3 strains per genotype. OMZ175 (serotype f) was belong to B genotype. Selected one of the A, B, C genotypings to sequenced and blasted. Then the results of the blastn showed that the identities of the gene sequence were 92%-98% between the serotype c and serotype f, part sequence of the serotype g was homology with the SrV+ of the serotype c, the protein sequence among serotype c, d, f, g were 77%-82%.

CONCLUSIONIt is reasonable to use some putative pipetides to study the anti-caries vaccine among the extended-V regions of the surface proteins in different serotype (c, d, f, g) in S. mutans.

DNA, Bacterial ; Genotype ; Membrane Proteins ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; Serogroup ; Streptococcus mutans

Objective To evaluate the enhanced magnification endoscopy in the diagnosis of Barrett esophagus,and to explore the relationship between mucosal surface patterns and pathological epithelial types of Barrett esophagus.Methods Enhanced magnification endoscopy was performed 'after spraying 2%-3% acetic acid on the surface of distal esophagus in 40 Barrett esophagus patients.Mucosal specimen were biop- syed.Results According to the mucosal types of Toyoda in 2003,there were three mucosal types:Ⅰ dot pat- tern 7(17.5%),5 of 7(71.4%)fundie type,Ⅱ reticular pattern 24(60.0%),16 of 24(66.7%)fundic type,Ⅲ cerebroid/villous 9(22.5%),intestinal metaplasia or dysplasia.Conclusion Enhanced magnifi- cation endoscopy helps to identify areas with intestinal metaplasia and dysplasia,and is useful in the diagno- sis of Barrett esophagus.

Objective To discuss the characteristic of brachial plexus root avulsion injury of high resolution MR imaging and the value in diagnosing of brachial plexus root avulsion injury early.Methods Fourty-five cases of brachial plexus root avulsion injury patients had being used for investigation to find the characteristic and diagnostic value of MR image of brachial plexus root avulsion injury,which all have pre-operative MR imaging and were diagnosed brachial plexus root avulsion injury by intra-operative exploration and electrophysiology form February 2006 to February 2011.Results Post-traumatic spinalmeningolceles were seen in 42 cases,the frequency was 93.3％; Displacement of spinal cord was seen in 25 cases,the frequency was 55.6％; Absence of anterior and posterior root of spinal nerve was seen in 8 eases,the frequency was 17.8％;Black line sign was seen in 18 cases,the frequency was 40.0％.The sensitivity,specificity,and accuracy of MRI in diagnosing brachial plexus root injury were 95.7％,77.8％ and 94.6％ respectively.Conclusion Posttraumatic spinalmeningolceles are most often seen in MR of brachial plexus root avulsion injury,this sign can play an important role in diagnosing and treatment of brachial plexus root avulsion injury.

BACKGROUNDThe Taq/B, Msp/ and I405V polymorphisms of cholesteryl ester transfer protein (CETP), an important regulatory factor of lipid metabolism, have been attracted much more attention by the researchers. In this study, we investigated the associations between these 3 polymorphisms of CETP gene and variations in plasma lipid and lipoprotein levels in patients with coronary heart disease (CHD).

METHODSGenomic DNA was extracted from leukocytes of 203 CHD patients and 100 control subjects using the salting out method. Genotyping of the CETP gene was performed using polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) techniques. Statistical analysis was conducted using the SPSS 10.0 software package.

RESULTSThe distribution of allele and genotype frequencies of the Taq/B, MspI, and I405V polymorphisms was similar in the CHD patient group and the control group. The B1B1 genotype of the Taq/B polymorphism was associated with significantly higher TC (P=0.039) and LDL-C (P=0.044) levels than the B2B2 genotype in CHD patients, and with significantly higher LDL-C (P=0.034) levels than the B2B2 genotype in controls. Homozygotes of the I405V polymorphism exhibited significantly higher HDL-C levels than VV homozygotes among control subjects (P=0.023). In male CHD patients with unambiguously assigned haplotypes, B2-M2-V/B2-M2-I patients demonstrated significantly higher HDL-C concentrations than B1-M2-V/B1-M2-I (P=0.023) and B1-M2-V/B1-M2-V patients (P=0.047).

CONCLUSIONSGenetic variations in the CETP gene may account for a significant proportion of the differences in plasma lipid and lipoprotein concentrations among the general population. The B1B1 genotype of the Taq/B polymorphism is probably a genetic risk factor for CHD in the study population.

Adult ; Aged ; Carrier Proteins ; genetics ; Cholesterol Ester Transfer Proteins ; Cholesterol, HDL ; blood ; Cholesterol, LDL ; blood ; Coronary Disease ; blood ; genetics ; Female ; Gene Frequency ; Glycoproteins ; genetics ; Humans ; Lipids ; blood ; Male ; Middle Aged ; Polymorphism, Genetic

OBJECTIVETo preliminarily investigate the relationship between the Lactate dehydrogenase (LDH) activity of Streptococcus mutans (S. mutans) and dental caries initiation.

METHODS100 S. mutans strains derived from caries-active and caries-free individuals were cultured in BHI medium supplemented with glucose. Cells were extracted and ruptured, and the extracted liquid protein was quantified with Coomassie brilliant blue G250 staining methods. LDH activity was assayed using the pyruvate-dependent oxidation of NADH-with and without FDP.

RESULTSLDH activity of the two groups strains had no difference (P > 0.05), but the distribution of differ class enzyme activity strains in the two groups was different (P < 0.05).

CONCLUSIONLDH activity of S. mutans is correlated to the initiation of dental caries to some extent. The measurement methods in this study can be applied in preliminary quantitation of LDH activity and the screening of S. mutans strains.

Case-Control Studies ; Dental Caries ; microbiology ; Humans ; Lactate Dehydrogenases ; Oxidation-Reduction ; Streptococcus mutans ; metabolism

Based our previous work, twelve purine derivatives were designed and synthesized as dual modulators of GPR119 and DPP-4by conjugating the GPR119 activating and DPP-4 inhibiting fragments with the position 6 and 9 of purine core via an approach of merged pharmacophores. Compound 11, bearing 2-fluoro-4-methylsulphonyl anilide and cyanopyrrolidine moieties, exhibited the most potent GPR119 agonistic activities (EC₅₀ = 0.33 μmol·L^-1, IA = 71.1%) and DPP-4 inhibitory (58.4% inhibition at 10 μmol·L^-1, 21.2% inhibition at 1 μmol·L^-1) activities in the in vitro antidiabetic study. Subsequently, we performed studies on structure activity relationships and molecular docking to guide the further drug design.

OBJECTIVETo isolate and purify a new phospholipase A2 (PLA2) homologue from Agkistrodon blomhoffii siniticus and investigate its effects on the gene expression profile of Hep3B cells.

METHODSThe PLA2 homologue was isolated and purified by reverse-phase high-performance liquid chromatography (HPLC) and its purity was determined also by HPLC. The relative molecular mass of the homologue was measured by electrospray ionization mass spectrum. The gene expression profile of Hep3B cells was detected with gene chip after exposure of the cells to 139 microg/ml PLA2 homologue for 12 h.

RESULTSThe purity of the PLA2 homologue was 97.2%, whose relative molecular mass was 13,900. After exposure of Hep3B cells to 139 microg/ml PLA2 homologue for 12 h, 19 genes were down-regulated and 20 up-regulated in the cells. The genes showing altered expressions in response to the exposure were mainly involved in cell cycle control and DNA damage repair, cell apoptosis and senescence, production of signal transduction molecules and transcription factors, cell adhesion, angiogenesis, and tumor invasion and metastasis.

CONCLUSIONSThe PLA2 homologue induces alterations in the expression of a wide variety of genes involved in the growth and metastasis of tumor cells. The results of this study provide clues for further study of the possible mechanism for the action of PLA2 homologue on Hep3B cells.

Agkistrodon ; Animals ; Carcinoma, Hepatocellular ; genetics ; Chromatography, High Pressure Liquid ; DNA Damage ; drug effects ; Gene Expression Profiling ; Gene Expression Regulation, Neoplastic ; drug effects ; Hyaluronan Receptors ; biosynthesis ; genetics ; Isoenzymes ; Liver Neoplasms ; genetics ; Phospholipases A ; isolation & purification ; pharmacology ; Phospholipases A2 ; Proto-Oncogene Proteins c-bcr ; biosynthesis ; genetics ; Snake Venoms ; enzymology ; Tumor Cells, Cultured

OBJECTIVETo analyze the changes of leptospirosis epidemic characteristics before and after the Phase 2 'reservoir store water project' in Chongqing section of the Three Gorges dam area and to provide prevention, control and intervention measures to prevent the spread of leptospirosis from infectious focus to the Three Gorges dam area and downstream region of Changjiang River.

METHODSChangshou district and Fengdu county were selected as surveillance sites. We monitored the source of infection through examining the serum antibody of patients, healthy groups together with farm cattle measured by micro agglutination test (MAT).

RESULTSporadic cases were reported before and after the storage of water in the reservoir. There was no significant difference found between mouse density before and after the Phase 2 reservoir project (chi2 = 1.00, P > 0.05). The main species of rat were Sewer rat before and Insectivorea after the storage of water. The germ-carrying rate of rats was 1.72% (10/583) and positive carrying rate of rats was 16.51% (18/109) when using PCR. Results showed a significant difference when comparing it to culture method (chi2 = 51.80, P < 0.01). Positive rate of leoptopirosis appeared in the serum of patients was 73.33% (33/45) with the major serum group as the Australia group. The rate of infection among the healthy group was 26.84% (233/868). There was significant difference seen between the serum antibody positive rate of epidemic prophase (23.85%) and epidemic anaphase (29.86%) of the healthy group (chi2 = 3.99, P < 0.05). The GMRT of ox serum antibody of leoptopirosis was 29.97 with Bailen group as the predominant microbial population.

CONCLUSIONThere was no epidemics of leptopirosis occurred in the Three Gorges dam area. There was no significant difference between mouse density before and after the storage of water in the reservoir. However, the major species of rats had a change. The natural infection level of people living in the dam area was low, but there existed potential of leoptopirosis outbreak.

Animals ; China ; Humans ; Leptospira ; genetics ; isolation & purification ; Leptospirosis ; epidemiology ; Polymerase Chain Reaction ; Population Surveillance ; Rats ; microbiology ; Rivers ; Water Supply