Objective To identify mutations site and clinical characteristics of a familial hypercholesterolemia(FH) proband diagnosed clinically through DNA sequencing and family analysis in the proband and his family members of 3 generations.Methods Blood samples and clinical data of the kindred of total 29 from 3 generations members were collected.Proband had a physical examination electrocar-diogrom and vascular ultrasound.The proband and his family members took routine clinical exams,and genomic DNA was isolated.The promoter region and the 18 exons of low density liporotein receptor(LDLR) gene were screened by Touch down polymerase chain reaction -single strand conformation polymorphism(PCR-SSCP) and DNA sequencing.The result of sequencing were matched gene sequence published in the BLAST database.Results 1.Increased intima-media thickness and plaque were detected in the common carotid artery,right subclavian artery of the proband.Aortic valve regurgitation was found by echocardiography.2.No mutation R3500Q of ApoB100 was observed.3.Two heterozygous mutations in exon 10 and 13 of LDLR gene (W462X and A606T) were identified.The proband and 5 members of paternal relatives showed W462X heterozygosis mutation in exon 10 of LDLR gene which introduced the change from tryptophone to a new stop codon.The proband's mother and grandmother harboured A606T heterozygous mutation in exon 13 of LDLR gene due to a single base pair substitution of G for A in the codon for residue 1 879.Conclusions Disease causing mutations of proband are W462X and A606T compound heterozygosis mutation in exon 10 and 13 of LDLR gene inherited from mother and father.Proband shows homozyous phenotype though the genotype analysis indicates heterozygous mutations.

Chronic Periodontitis ; complications ; diagnosis ; diagnostic imaging ; therapy ; Dental Pulp Diseases ; complications ; diagnosis ; diagnostic imaging ; therapy ; Dental Scaling ; Female ; Humans ; Periodontal Debridement ; Periodontal Diseases ; complications ; diagnosis ; diagnostic imaging ; therapy ; Radiography ; Root Canal Therapy ; Root Planing ; Young Adult

Candida shehatae xyl1 gene and Pichia stipitis xyl2 gene were amplified by PCR and the xyl1 and xyl2 were both placed under the promoter GAL of vector pYES2 to produce the recombinant expression vector pYES2-P12. Subsequently the pYES2-P12 vector was transformed into S. cerevisiae YS58 by LiAc to produce the recombinant yeast YSS8-12. It was indicate that the recombinant yeast YSS8-12 could converse xylose to ethanol with the xylose consumption rate of 81. 3%.

There are abundant of lignocelluloses in agricultural wastes. And it can provide a variety of sugars involve pentose and hexose through pretreatments or hydrolyzation of these lignocelluloses. This review summarized the current status of bioethanol production by bacteria, and compared the productivity of some kinds of ethanologenic bacteria such as recombinant Zymomonas mobilis and Escherichia coli.

Objective To study on the metal clip installation to avoid post-operative bleeding in pa- tients accepted papilla sphinctecotomy.Methods One hundred and eighty five patients who accepted ERCP +EST were divided into two groups:Group 1 was given routine regimen alone(N=95),group 2,given routine regimen and metal clip to prevent post-operative bleeding.Results The postoperative bleeding hap- pened in 3(3.2%)cases of Group 1 and none in Group 2,there is significant difference between these two groups(P＜0.05).The breeding cases in group 1 were controlled by metal clip under endoscopy successful- ly.Conclusion Preventive usage of metal clip was significantly decreased the incidence of post-operative bleeding in EST patients.

OBJECTIVETo study the prevalence of Tannerella forsythensis (T. forsythensis) in the subgingival plaque of type 2 diabetes, analyze the relationship between Tforsythensis and related factors, discuss the role of T. forsythensis in the chronic periodontitis and type 2 diabetes.

METHODS160 subgingival plaque samples were collected from type 2 diabetic patients and pathogens genomic DNA were extracted by phenol and chloroform from plaque. T. forsythensis was detected by polymerase chain reaction, and Pearson correlation and Logistic regression analysis were used to analyze the relationship between T. forsythensis and systemic factors and periodontal status.

RESULTSThe prevalence of T. forsythensis in mild, moderate, severe periodontitis group was 47.82%, 48.71%, 67.39% respectively, and the prevalence was higher in the severe periodontitis group than in mild, moderate group (P < 0.05). There was no T. forsythensis in 6 diabetic patients with healthy periodontium. Logistic regression analysis illustrated that the prevalence of T. forsythensis was associated with simplified oral hygiene index (OHI-S) and diabetic duration (OR = 1.947, OR = 0.873).

CONCLUSIONThe prevalence of T. forsythensis in type 2 diabetes with chronic periodontitis was related with oral hygiene, periodontal status and diabetic duration.

Aged ; Bacteroides ; Chronic Periodontitis ; Dental Plaque ; Diabetes Mellitus, Type 2 ; Humans ; Periodontitis ; Polymerase Chain Reaction ; Porphyromonas gingivalis ; Prevalence

OBJECTIVETo establish the model of cellular inflammatory responses of gingival epithelial cells in vitro induced by Porphyromonas gingivalis vesicle and to probe into the pathogenesis of Porphyromonas gingivalis in periodontitis.

METHODSThe effect of Porphyromonas gingivalis vesicle on prostaglandin E(2) (PGE(2)) production of gingival epithelial cells was detected by ELISA and the effects of Porphyromonas gingivalis vesicle on cyclooxygenase-2 (COX-2), interleukin (IL)-6 and IL-8 mRNA expression in gingival epithelial cells were determined by Real-time reverse transcription-polymerase chain reaction (RT-PCR).

RESULTSPorphyromonas gingivalis vesicle dose-dependently induced PGE(2) production and up-regulated COX-2, IL-6 and IL-8 mRNA expression in gingival epithelial cells significantly.

CONCLUSIONSCellular inflammatory responses of gingival epithelial cells induced by Porphyromonas gingivalis vesicle may contribute to the initiation and progression of periodontitis.

Bacterial Adhesion ; Cells, Cultured ; Cyclooxygenase 2 ; immunology ; metabolism ; Dinoprostone ; immunology ; metabolism ; Epithelial Cells ; immunology ; metabolism ; microbiology ; Gingiva ; immunology ; metabolism ; microbiology ; Humans ; Interleukin-6 ; immunology ; metabolism ; Interleukin-8 ; immunology ; metabolism ; Porphyromonas gingivalis ; immunology ; pathogenicity

OBJECTIVETo assess the prevalence of specific kgp genotypes in puberty gingivitis and investigate their possible association with disease severity.

METHODSSubgingival plaque samples were collected from 72 pubertal children aged from 14 to 17 years, which were divided into two groups, gingivitis group and healthy (control) group. Clinical parameters were recorded beforehand. PCR technique was used to amplify the region encoding the catalytic domain of gingipain K (KGP). The PCR products were digested with restriction enzymes Mse I.

RESULTSThe kgp-A genotype was digested in fragments of 102 bp, 288 bp and 402 bp, and kgp-B genotype was unrestricted with 792 bp. Virulent strain P. gingivalis W83 was manifested by kgp-A genotype while a virulent strain P. gingivalis ATCC 33277 was manifested by kgp-B genotype. All P. gingivalis positive subjects were subgingivally colonized by only one kgp genotype. The prevalence of kgp-A genotype in puberty gingivitis group and gingival healthy group was 79.0% and 22.2% respectively. The distribution differences of genotypes between the two groups were statistically different (P = 0.028). There was no statistically significant difference in the clinical parameters between pubertal subjects harboring P. gingivalis of kgp-A genotype and kgp-B genotype (P > 0.05).

CONCLUSIONSMost subjects with puberty gingivitis were harbored by the same kgp genotype as that of virulent strain P. gingivalis W83. It may be necessary to continue to monitor individuals who are positive for P. gingivalis of kgp-A genotype since their risk of developing periodontal diseases may be increased in the future.

Adolescent ; Case-Control Studies ; Dental Plaque ; microbiology ; Female ; Genotype ; Gingivitis ; microbiology ; Humans ; Male ; Polymerase Chain Reaction ; Porphyromonas gingivalis ; genetics

OBJECTIVETo detect and compare the activity and intensity of gingipain K (Kgp)-caspase like subdomain in culture medium and cell extract of Porphyromonas gingivalis (Pg) isolates in puberty gingivitis and to reveal the possible association of Kgp with puberty gingivitis.

METHODSThirty-six children of 14 to 17 years old were enrolled in this study. Clinical parameters including gingival index (GI), sulcus bleeding index (SBI) and probing depth (PD) were evaluated. Subgingival plaque samples were collected and Pg isolates were obtained. 16S rRNA PCR was used to confirm Pg clinical isolates. Bacteria were grown in batches of BHI base and harvested at the end of log-phase growth. Culture fractions (culture medium and cell extract) of 10 Pg isolates were performed with SDS-PAGE and Western blot technique using primary antibody against specific Kgp-caspase like subdomain. Activity of Kgp in both samples was detected as well. The data were statistically analyzed using SPSS 11.5 software. The relationship between the Kgp intensity/activity of Kgp and the clinical parameters was statistically analyzed using Spearman correlation coefficient.

RESULTSThere was positive correlation between the intensity/activity of Kgp and the clinical parameters (P < 0.05).

CONCLUSIONSThe Kgp in clinical isolates of Pg from puberty gingivitis is in complicated forms. Caspase-like molecules with low molecular weight may exist as intracellular functional protein molecules which can affect the interaction between Pg and host. Kgp was contributes in certain degree to the pathogenesis of puberty gingivitis.

Adhesins, Bacterial ; genetics ; metabolism ; Adolescent ; Cysteine Endopeptidases ; genetics ; metabolism ; Dental Plaque ; microbiology ; Female ; Gingivitis ; enzymology ; microbiology ; Humans ; Male ; Porphyromonas gingivalis ; genetics ; isolation & purification ; metabolism

OBJECTIVETo identify the differential genes in Porphyromonas gingivalis (P.gingivalis) highly toxic strain W83 and minimally toxic strain ATCC 33277.

METHODSUsing suppression subtractive hybridization (SSH) to compare P.gingivalis highly toxic strain W83 (tester) and minimally toxic strain ATCC 33277 (driver). The chromosomal DNAs were purified from P.gingivalis W83 and P.gingivalis ATCC 33277, and digested by restriction enzyme RsaI. The tester DNA samples were separated and ligated with adaptor 1 and adaptor 2R. Two subtractive hybridization and PCR profile were performed. Tester-specific DNAs also were selectively amplified. The mixture of subtracted DNA fragments were ligated with pMD-18T vector and transformed to competent cells E.coli JM109. The differential subtraction library was established. The positive clones were identified by PCR and then sequenced, and searched homologically.

RESULTSSubtractive library which had high subtractive efficiency was successfully set up and 36 positive clones were screened by SSH. The fragments from 88 bp to 372 bp were enriched in P.gingivalis highly toxic strain W83 sequences which were absent from P.gingivalis ATCC 33277. Through dot blot analysis confirmed that all these fragments were present in P.gingivalis W83 but absent from ATCC 33277. The GenBank homology search indicated that among them, several genes were associated with two paralogous regions of the chromosome; Some genes are associated with evasion of P.gingivalis W83; Another gene was related to antibiotic resistance and the products of some genes were virulence and acquisition of peptides.

CONCLUSIONSComparative whole-genome analysis of highly toxic and minimally toxic strains of P.gingivalis has identified the clustering of genes that are present in W83 but divergent in or absent from ATCC 33277. These genes may provide an important clue for studying the mechanism of occurrence and development of periodontal disease.

Genes, Bacterial ; Genome, Bacterial ; Nucleic Acid Hybridization ; Porphyromonas gingivalis ; genetics ; pathogenicity ; Sequence Analysis, DNA ; Virulence ; genetics