BACKGROUND: An accurate and rapid method for specise identification of coagulase negative staphylococci(CNS) has been increasingly necessary for the clinical significance and planning the management of patients with staphylococcal infections. Recently, it has been reported that there is a highly conserved area on their 60KDa heat shock protein(HSP60) gene sequences between the interspecies of CNS and it can be amplified by a set of universal degenerate primer. This led us our attention to focus on whether the PCR-based RFLP method using Mse / restriction enzyme could be a useful tool for the species identification of CNS. METHODS: In the present study, we performed PCR-based RFLP analysis using a set of degenerate primers covering HSP60 and Mse / restriction enzyme on the reference strains and 25 clinical isolates(10 of S. epidermidis, 10 of S. haemolyticus, 4 of S. lugdunensis and 1 of S. warneri) which were previously identified by the API-STAPH, Vitek GPI card and/or with conventional biochemical test. RESULT: All the seven reference strains revealed that each strain has a distinct electrophoresed band patterns with combination of different number (up to 8) and size of fragments. And these distinct band patterns showed remarkable concordance with the seven reference strains and 25 clinical isolates. CONCLUSION: These result strongly suggest that the PCR-RFLP method using degenerate primers covering the HSP60 gene and Mse / digestion enzyme offer a convenient and accurate tool for species-specific identification of CNS.
Chaperonin 60* ; Coagulase* ; Digestion ; Heat-Shock Proteins* ; Hot Temperature* ; Humans ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; Shock ; Staphylococcal Infections

OBJECTIVETo identify the single nucleotide polymorphisms (SNPs) in the regulatory and coding regions of heat shock protein 60 gene and search for its genetic makers in Chinese Han people.

METHODSThe 5' flank region, parts of the exons and introns of hsp60 gene were resequenced to identify the SNPs in Chinese Han people, and then the sequenced results to the Japanese, European and African's data in National Center for Biotechnology Information (NCBI) and HapMap databases were compared.

RESULTSOne novel SNP was identified in exon 2 resulting in synonymous variant and the G allele frequency was 0.025. There were 11 reported SNPs in the sequenced region. The minor allele frequencies of rs1116734, rs3749095, rs1050347, rs8539 were 0.51, 0.30, 0.29, 0.49. The heterozygosity of the other 7 SNPs was 0. The distributions of rs1116734, rs1050347, rs8539, rs3749095 in Chinese Han people were similar to the Japanese's. The hsp60 rs3749095 which was not found in Japanese people was a high-frequency SNP in Chinese Han people; the distribution of rs8539 in Chinese Han people was quite different from European and African's (P < 0.01).

CONCLUSIONThe SNPs of hsp60 in Chinese Han people are different from the other peoples. The SNPs of hsp60 gene rs1116734, rs3749095, rs1050347, rs8539 are very common in Chinese Han people and might be used for candidate genetic markers of hsp60 gene.

Adolescent ; Adult ; Alleles ; Chaperonin 60 ; genetics ; China ; ethnology ; Exons ; genetics ; Female ; Gene Frequency ; Humans ; Male ; Polymorphism, Single Nucleotide

As thermostable enzymes and organisms are much more needed, researches on heat shock proteins(HSPs) of hyperthermophilic archaea have drawn more concerns. HSPs from hyperthermophilic archaea are concise only with HSP60, sHSP, prefoldin and AAA+proteins, but without HSP100s, HSP90s, HSP70 (DnaK), HSP40 (DnaJ) and GrpE which are common in mesophilic or thermophilic archaea. Accordingly, studies on the structure, function and operation mechanism of these four groups are much more important and meaningful. This review focuses on the recent progress in the researchs on the structure, function, operation mechanism and cooperation of the HSPs from hyperthermophilic archaea. The problems and obfuscations in these HSPs are analyzed, and farther research direction and key points are put out.
Archaea ; classification ; metabolism ; Archaeal Proteins ; metabolism ; Chaperonin 60 ; metabolism ; Heat-Shock Proteins ; genetics ; metabolism ; Molecular Chaperones ; metabolism

OBJECTIVE: To investigate the changes of creatine kinase-MB (CK-MB) and heat shock protein 60 (HSP 60) in rats without electric marks after electric injury, to identify the relationship of the CK-MB, HSP 60 and the time of electric injuries, and to evaluate the damage to cells after electric injury. METHODS: The animal model of electric injury without electric marks was established by alternating current (voltage 110 V). Automatic biochemistry analyzer was used to detect the serum CK-MB and immunohistochemical staining technology was used to analyze the tissues of myocardium and left lobe of liver. RESULTS: The amount of serum CK-MB was increased when the rats were injuried, and reached the peak at 30min. Then the amount of CK-MB began to decrease and showed a slight downward trend in 3-5 h after electric injury, and leveled off at 6 h. Immunohistochemistry staining also showed the changes of HSP 60 of rats' myocardial cells and hepatic cells regularly after electric injury. CONCLUSION The regular changes of serum CK-MB and tissular HSP 60 in rats can be used to diagnosis electric injury and assess the injury of internal organs after the electric injury without electric marks.
Animals ; Chaperonin 60/metabolism* ; Creatine Kinase, MB Form/metabolism* ; Electric Injuries/complications* ; Immunohistochemistry ; Liver/pathology* ; Myocardium/pathology* ; Rats

Toxoplasma gondii can infect all the vertebrates including human, and leads to serious toxoplasmosis and considerable veterinary problems. T. gondii heat shock protein 60 (HSP60) is associated with the activation of antigen presenting cells by inducing initial immune responses and releasing inflammatory cytokines. It might be a potential DNA vaccine candidate for this parasite. A pVAX-HSP60 DNA vaccine was constructed and immune responses was evaluated in Kunming mice in this study. Our data indicated that the innate and adaptive immune responses was elicited by successive immunizations with pVAX-HSP60 DNA, showing apparent increases of CD3e+CD4+ and CD3e+CD8a+ T cells in spleen tissues of the HSP60 DNA-immunized mice (24.70±1.23% and 10.90±0.89%, P < 0.05) and higher levels of specific antibodies in sera. Furthermore, the survival period of the immunized mice (10.53±4.78 day) were significantly prolonged during the acute T. gondii infection. Decrease of brain cysts was significant in the experimental group during the chronic infection (P < 0.01). Taken together, TgHSP60 DNA can be as a vaccine candidate to prevent the acute and chronic T. gondii infections.
Animals ; Antibodies ; Antigen-Presenting Cells ; Brain ; Chaperonin 60 ; Cytokines ; DNA ; Humans ; Immunization ; Mice ; Parasites ; Spleen ; T-Lymphocytes ; Toxoplasma ; Toxoplasmosis ; Vertebrates

OBJECTIVE: To investigate the effects of antibodies to HSP 60 on the early mouse embryo development in vitro. METHODS: The 175 late 2-cell mouse embryos were obtained from 6-7 week old female ICR mice. 5-10 embryos were placed in each well. The embryos were incubated in the Ham's F-10 medium supplemented with 100 microgram/mL of monoclonal antibody to HSP 60 (66), monoclonal mouse IgG1 (55), and medium alone (54), respectively, at 37degrees C in a 5% CO2 humidified air chamber, and mouse embryo developments were observed daily. RESULTS: On day4, growth arrests were more prominent in anti-HSP 60 containing group compared to IgG1 containing group, medium only group (0% vs 16%, 14%), and these results were statistically significant (p=0.0032). Especially those inhibitory effects were observed in early stage of embryo development (day1) and these results were also statistically significant (31% vs 83%, 77%, p<0.0001). Moreover, we found out that cellular degenerations were more common in anti-HSP 60 containing group and this features were prominent on day2. CONCLUSION: Anti-HSP 60 elicited a strong growth inhibitory and degenerative effect on early mouse embryo development. These findings suggest that HSP 60 may exert a protective effect against mouse embryo degeneration or apoptosis.
Animals ; Antibodies ; Antibodies, Monoclonal* ; Apoptosis ; Chaperonin 60* ; Embryonic Development* ; Embryonic Structures* ; Female ; Heat-Shock Proteins* ; Hot Temperature* ; Humans ; Immunoglobulin G ; Mice* ; Mice, Inbred ICR ; Pregnancy

OBJECTIVETo investigate the value of grOEL gene sequence in phylogenetic analysis and typing of Salmonella.

METHODSThe grOEL gene was amplified by PCR, sequenced and analyzed using Bioedit and DNAstar software. The Salmonella strains were identified using PCR-restriction fragment length polymophism (PCR-RFLP).

RESULTSThe conservative and variable regions of grOEL gene of Salmonella serogroup were separately distributed and most of the small mutant regions distributed intermittently among the conservative regions. The phylogenetic tree of Salmonella based on the nucleotides differed from that generated based on the amino acid sequence. O8, O9 and O10 had the closest consanguinity, and 5 patterns were identified by PCR-RFLP.

CONCLUSIONThe grOEL gene can be used as a genetic marker for phylogenetic analysis of Salmonella and also as a target sequence for Salmonella typing identification.

Bacterial Typing Techniques ; Chaperonin 60 ; genetics ; DNA, Bacterial ; genetics ; Phylogeny ; Polymerase Chain Reaction ; methods ; Polymorphism, Restriction Fragment Length ; Salmonella ; classification ; genetics ; Sequence Analysis, DNA

To explore the roles of tumor necrosis factor-alpha (TNF-alpha) and heat shock protein 60 (HSP-60) in women with tubal factor infertility (TFI) associated with Chlamydia trachomatis, and to determine the mechanisms of fallopian adhesions in Chlamydia trachomatis (CT) infections, the expressions of TNF-alpha and HSP-60 were quantitatively determined in 60 cases of TFI and 30 controls by immunohistochemical technique. The patients with TFI were further divided into group A and group B according to the CT-DNA of cervical specimens of PCR. The quantitative analysis was conducted by employing computerized image analysis system. It is found that the expressions of TNF-alpha and HSP-60 were much higher in TFI patients than those of controls. Among CT-HSP responders, a stronger expression was correlated with more severe salpingeal pathology. It is concluded that TNF-alpha and HSP-60 play very important roles in fallopian tube adhesion and occlusion in TFI due to CT infection.
Adult ; Chaperonin 60 ; blood ; Chlamydia Infections ; blood ; Chlamydia trachomatis ; Fallopian Tube Diseases ; complications ; virology ; Female ; Humans ; Infertility, Female ; etiology ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVE: To compare the distribution and expression of hsp60 and hsp27 in placental trophoblast between preterm and term placenta and to observe hsp immune response in relation to the pathogenesis of preterm birth. METHODS: 22 cases of preterm trophoblast, between 24 weeks and 36 weeks gestation, which were developed spontaneous onset or less than 24 hours after rupture of membrane were obtained. And aged-matched, 22 cases of normal term trophoblast, as control were also obtained after informed consent from each patient. The protein extraction form trophoblast was stained by immunohistochemical methods and was measured by the assay of Western blots. And the density of band using Image-writer were taken and statistical assay were performed as significance <0.05. RESULTS: The expressions of hsp60 and hsp27 in trophoblast of preterm and term placenta were identified by immunohistochemical staining method. The hsp60 had significantly higher expression in trophoblast of preterm birth than in that of term birth (P<0.001) and the hsp27 also had significantly higher expression in trophoblast of preterm birth than in that of term birth (P<0.02) CONCLUSION: The higher expression of hsp60 and hsp27 in trophoblast of preterm birth might be suggested the development of immune response to occur preterm labor Further study are necessary to determine the exact actions of hsp60 and27 in trophoblast and to understand the immune mechanism of preterm birth.
Blotting, Western ; Chaperonin 60 ; Female ; Heat-Shock Proteins ; Hot Temperature ; Humans ; Informed Consent ; Membranes ; Obstetric Labor, Premature ; Placenta ; Pregnancy ; Premature Birth ; Rupture ; Term Birth ; Trophoblasts

PURPOSE: The aim of this study was to define the immunoreactive specificity of Porphyromonas gingivalis (P. gingivalis) heat shock protein (HSP) 60 in periodontitis and atherosclerosis. METHODS: In an attempt to define the cross-reactive bacterial heat-shock protein with human self-antigen at molecular level, we have introduced a novel strategy for cloning hybridoma producing anti-P. gingivalis HSP 60 which is polyreactive to bacterial HSPs or to the human homolog. RESULTS: Five cross-reactive clones were obtained which recognized the #19 peptide (TLVVNRLRGSLKICAVKAPG) among 37 synthetic peptides (20-mer, 5 amino acids overlapping) spanning the whole molecule of P. gingivalis HSP 60. We have also established three anti-P. gingivalis HSP 60 monoclonal antibodies demonstrating mono-specificity. These clones recognized the #29 peptide (TVPGGGTTYIRAIAALEGLK). CONCLUSIONS: Peptide #19 and #29 of P. gingivalis HSP 60 might be important immunoreactive epitopes in the immunopathogenic mechanism of bacterial antigen-triggered autoimmune diseases.
Amino Acids ; Antibodies ; Antibodies, Monoclonal ; Autoimmune Diseases ; Chaperonin 60 ; Clone Cells ; Cloning, Organism ; Epitopes ; Heat-Shock Proteins ; Humans ; Hybridomas ; Peptides ; Periodontitis ; Porphyromonas ; Porphyromonas gingivalis ; Sensitivity and Specificity