ABSTRACT
Dental caries is the most common oral disease and the most common cause of resin restorations. In minimally invasive dentistry, the principle behind cavity preparation is to remove external caries-infected dentin (CID) and preserve internal caries-affected dentin (CAD) and sound dentin (SD). The cavity floor is mainly composed of CAD, but the poor bonding performance of CAD has become a widespread concern. This study evaluated the performance of a new collagen-reactive monomer (ITCM) used as a primer to improve the bonding performance of CAD. The experimental specimens were grouped as follows: SD, CAD, and ITCM-pretreated CAD (CAD-ITCM). Dentin slices were obtained for attenuated total reflectance-Fourier transform infrared (ATR-FTIR) analysis. The bonded samples were subjected to microtensile bond strength analysis after 24 h of water storage or aging by thermocycling, and the bonding interface quality was evaluated by nanoleakage assessment, interfacial nanoindentation testing, and in situ zymography. Cytotoxicity experiments with ITCM were performed. ATR-FTIR showed that the isocyanate groups in ITCM can covalently bind and form hydrogen bonds with the collagen in CAD to mediate chemical bonding. ITCM pretreatment significantly improved the bond strength of CAD (P < 0.05), reduced interfacial nanoleakage, improved the sealing of the bonding interface, enhanced the homogeneity of the hybrid layer, and inhibited matrix metalloproteinase activity. In addition, ITCM presented acceptable biocompatibility for dental restorative application. Taken together, this study reported the application of ITCM to induce collagen-based chemical bonding in the CAD bonding system, which fills the gap in strategies to improve the bonding performance of CAD immediately and after aging and has important clinical application prospects.
Subject(s)
Dental Bonding , Dental Caries , Humans , Composite Resins/chemistry , Dentin-Bonding Agents/chemistry , Dental Caries Susceptibility , Resin Cements/chemistry , Dentin , Tensile Strength , Acid Etching, Dental , Materials Testing , CollagenABSTRACT
Dentin bonding is a dynamic process that involves the penetration of adhesive resin monomers into the extrafibrillar and intrafibrillar demineralized collagen matrix using a wet-bonding technique. However, adhesive resin monomers lack the capacity to infiltrate the intrafibrillar space, and the excess water that is introduced by the wet-bonding technique remains at the bonding interface. This imperfectly bonded interface is inclined to hydrolytic degradation, severely jeopardizing the longevity of bonded clinical restorations. The present study introduces a dentin bonding scheme based on a dry-bonding technique, combined with the use of extrafibrillar demineralization and a collagen-reactive monomer (CRM)-based adhesive (CBA). Selective extrafibrillar demineralization was achieved using 1-wt% high-molecular weight (MW) carboxymethyl chitosan (CMCS) within a clinically acceptable timeframe to create a less aggressive bonding substance for dentin bonding due to its selectively extrafibrillar demineralization capacity. CMCS demineralization decreased the activation of in situ collagenase, improved the shrinking resistance of demineralized collagen, and thus provided stronger and more durable bonding than traditional phosphoric acid etching. The new dentin bonding scheme that contained CMCS and CBA and used a dry-bonding technique achieved an encouraging dentin bonding strength and durability with low technical sensitivity. This bonding scheme can be used to improve the stability of the resin-dentin interface and foster the longevity of bonded clinical restorations.
ABSTRACT
When damaged or fractured collagen-rich hard tissues are repaired by resin material, the collagen matrix may be used as a scaffold, after removal of the natural minerals, for resin monomers to penetrate and polymerize in-situ. Formation of a collagen-polymer hybrid biocomposite via mechanical hybridization provides a stable and strong link between endogenous tissue and the prosthesis for successful clinical integration. However, the heterogeneity between hydrophobic resin polymers and hydrophilic collagen presents a challenge to the quality of hybrid biocomposite. The objective of the present study was to evaluate the potential benefits of a collagen-reactive monomer (CRM, an isocyanate-terminated urethane-based methacrylate) with covalent affinity to collagen as "chemical link" to enhance in-situ resin hybridization within a collagen scaffold. Here, the CRM ligand with active isocyanate group may be chemically grafted onto the collagen receptor via covalent and hydrogen bonds. Dentin-derived collagen chemical modified by CRM shows improved mechanical property, thermostability and enzymatic stability. Moreover, CRM inhibited both exogenous and endogenous collagenase activities. The modification of collagen by chemical grafting of resin monomers improved its mechanical and physicochemical properties and demonstrated the potential of CRM for use in promoting chemical adhesion and creating a much stronger and durable bonding interface. Formation of a chemical bond between polymer and collagen scaffold in-situ improves the mechanical performance of collagen and may create a much stronger and durable collagen-polymer hybrid material. Addition of CRM into adhesives might effectively prolong the longevity of clinical resin-bonded restorations.
Subject(s)
Collagen/chemistry , Composite Resins/chemistry , Isocyanates/chemistry , Methacrylates/chemistry , Urethane/chemistry , Binding Sites , Collagen/metabolism , Collagenases/chemistry , Collagenases/metabolism , Composite Resins/metabolism , Dentin/chemistry , Dentin/metabolism , Materials Testing , Molecular Docking Simulation , Temperature , Tensile StrengthABSTRACT
This study evaluated the use of a new collagen-reactive monomer (CRM), isocyanate-terminated urethane methacrylate precursor, which has covalent affinity to dental collagen, in the formation of dentin-resin bonds and compared it with 2 other dental adhesives. Dentin specimens were bonded with either the CRM-based adhesive (CBA), One-Step (OS; Bisco, Inc.), or a negative adhesive (NA) control and subjected to 24-h storage in water, thermocycling to simulate 1-y clinical function, or a matrix metalloproteinase-mediated aging process. We tested the microtensile bond strength (µTBS), characterized the bonding interface with an atomic force microscope, conducted micro-Raman analysis, and performed leakage tests and in situ zymography. CBA and OS exhibited comparable bonding strength after 24 h (P > 0.05); however, there was a sharp decrease in µTBS after aging for all except CBA (P < 0.001). Raman spectra results indicated increased collagen crosslinking and chemical reaction between the adhesive and collagen in the CBA group. CBA achieved high-quality hybridization with collagen, improving mechanical properties and integrity, and decreased the enzyme-mediated degradation of the bonding interface by inhibiting collagenolytic activity. With the promising bonding durability of coapplied CBA, CRM may be the first dental adhesive to provide strong and long-lasting resin-dental collagen bonding without the additional conditioning step. The use of CBA results in high-quality hybrid layers that protect the resin-dentin interface from harmful biological and chemical activities commonly occurring in the oral environment.
Subject(s)
Dental Bonding , Collagen , Dentin , Dentin-Bonding Agents , Materials Testing , Resin Cements , Surface Properties , Tensile StrengthABSTRACT
Objective: To evaluate the cost-benefit and cost-effectiveness of current strategy for preventing mother-to-child transmission (PMTCT) of hepatitis B virus. Methods: A decision tree model with the Markov process was developed and simulated over the lifetime of a birth cohort in Zhejiang Province in 2016. The current PMTCT strategy was compared with universal vaccination and non-vaccination. Costs were assessed from social perspective. Benefits were the savings from reduced costs associated with disease and effectiveness were measured by quality-adjusted of life-years (QALY) gained. The net present value (NPV), cost-benefit ratio (BCR) and incremental cost-effectiveness ratio (ICER) were calculated. Univariate and Probabilistic Sensitivity Analyses (PSA) were performed to assess parameter uncertainties. The parameters of costs and utilities value of hepatitis B-related disease came from the results of the field survey, which were obtained by face-to-face questionnaire survey combined with inpatient medical records, including eight county and municipal hospitals in Jinhua, Jiaxing and Taizhou. A total of 626 outpatients and 523 inpatient patients were investigated. The annual total costs of infection was calculated by combining the costs of outpatient and inpatient. Results: The PMTCT strategy showed a net-gain as 38 323.78 CNY per person, with BCR as 21.10, which was higher than 36 357.80 CNY per person and 13.58 respectively of universal vaccination. Compared with universal vaccination, the PMTCT strategy would save 2 787.07 CNY per additional QALY gained for every person, indicating that PMTCT would be cost-saving. The most important parameters that could affect BCR and ICER were the vaccine coverage rate and costs of hepatitis B related diseases respectively. The PSA showed the PMTCT strategy was preferable as it would gain more QALY and save costs. Conclusions: The PMTCT strategy appeared as highly cost-beneficial and highly cost-effective. High vaccination rate was a key factor of high economic value.
Subject(s)
Hepatitis B Vaccines/economics , Hepatitis B/prevention & control , Infectious Disease Transmission, Vertical/prevention & control , Vaccination/economics , China , Cost-Benefit Analysis , Female , Hepatitis B/economics , Hepatitis B/transmission , Hepatitis B Vaccines/administration & dosage , Humans , Infant , Infectious Disease Transmission, Vertical/economics , Pregnancy , Quality-Adjusted Life Years , Vaccination/statistics & numerical dataABSTRACT
From 2014 to 2015, four novel highly pathogenic PRRS virus (HP-PRRSV) strains named 14LY01-FJ, 14LY02-FJ 15LY01-FJ, and 15LY02-FJ were isolated from high morbidity (100%) and mortality (40%-80%) in piglets and sows in Fujian Province. To further our knowledge about these novel virus strains, we characterized their complete genomes and determined their pathogenicity in piglets. Full-length genome sequencing analysis showed that these four isolates were closely related to type 2 (North American type, NA-type) isolates, with 88.1%-96.3% nucleotide similarity, but only 60.6%-60.8% homology to the Lelystad virus (LV) (European type, EU-type). The full length of the four isolates was determined to be 15017 or 15018 nucleotides (nt), excluding the poly(A) tail. Furthermore, the four isolates had three discontinuous deletions (aa 322-432, aa 483, and aa 504-522) within hypervariable region II (HV-II) of Nsp2, as compared to the reference strain VR-2332. This deletion pattern in the four isolates is consistent with strain MN184 and strain NADC30 isolated from America. Phylogenetic and molecular evolutionary analyses indicated that these virulent strains originated from a natural recombination event between the JXA1-like HP-PRRSV (JXA-1 is one of the earliest Chinese HP-PRRSV strains; sublineage 8.7) and the NADC30-like (lineage 1) PRRSV. Animal experiments demonstrated that these four strains caused significant weight loss and severe histopathological lung lesions as compared to the negative control group. High mortality rate (40% or 80%) was found in piglets infected with any one of the four strains, similar to that found with other Chinese HP-PRRSV strains. This study showed that the novel variant PRRSV was HP-PRRSV, and it is therefore critical to monitor PRRSV evolution in China and develop a method for controlling PRRS.
Subject(s)
Porcine Reproductive and Respiratory Syndrome/virology , Porcine respiratory and reproductive syndrome virus/pathogenicity , Swine Diseases/virology , Amino Acid Sequence , Animals , Base Sequence , China/epidemiology , Female , Phylogeny , Porcine Reproductive and Respiratory Syndrome/epidemiology , Porcine respiratory and reproductive syndrome virus/genetics , Porcine respiratory and reproductive syndrome virus/isolation & purification , Random Allocation , Sequence Alignment/veterinary , Swine , Swine Diseases/epidemiology , Viral Nonstructural Proteins/genetics , VirulenceABSTRACT
Porcine circovirus type 2 (PCV2) is considered to be the main pathogen in PC-associated diseases, and significantly affects the global pig-producing industry. PCV2 continuously evolves by point mutations and genome recombinations. In the present study, we aimed to further identify recombinant PCV2 strains. We used polymerase chain reaction to detect PCV2 in the carcasses of pigs with suspected infections from different regions of Guangdong Province in China. DNA was extracted from samples with confirmed infection and full- genome amplification, sequencing, phylogenetic tree construction, gene recombination detection, and sequence alignment were performed in gene recombination analysis. Our results show that recombination occurred between the strains SHC (DQ104421) and ZhuJi2003 (AY579893). The recombination resulted in three recombinants: GD003 (KM503044), GD005 (KM487708), and GD008 (KM487709). Further analyses revealed that these novel recombinants appeared to result from recombination between the PCV2a and PCV2b strains, with crossover regions located in ORF2. This study was a comprehensive analysis that used several different methods, which demonstrated that a cluster of PCV2 strains resulted from the same type of inter-genotypic recombination pattern, with a breakpoint in the structural protein coding region. The results of our study provide both information on the recombination mechanism and disease pathogenesis and useful data for the prevention of PCV2 in the swine industry.
Subject(s)
Circoviridae Infections/virology , Circovirus/genetics , DNA, Viral/genetics , Reassortant Viruses/genetics , Recombination, Genetic , Animals , Base Sequence , Cell Line , Circoviridae Infections/pathology , Circovirus/classification , Circovirus/pathogenicity , Epithelial Cells/pathology , Epithelial Cells/virology , Lymph Nodes/pathology , Lymph Nodes/virology , Molecular Sequence Data , Phylogeny , Reassortant Viruses/pathogenicity , Sequence Alignment , Spleen/pathology , Spleen/virology , SwineABSTRACT
We have shown earlier that overexpression of Calreticulin (CRT) contributed to a poor prognosis for patients with esophageal squamous cell carcinoma (ESCC). Here, we have shown an important role of CRT in tumorigenesis through enhancing cell motility and anoikis resistance. SiRNA-mediated knockdown of CRT caused impaired cell migration, invasion and resistance to anoikis. Notably, CRT downregulation decreased the expression of Cortactin (CTTN), which has been previously reported as a candidate oncogene associated with anoikis through the PI3K-Akt pathway. In addition, Akt phosphorylation was abolished after CRT downregulation and its activation can be refreshed by CRT upregulation, suggesting that CRT-enhanced cell resistance to anoikis through the CRT-CTTN-PI3K-Akt pathway. Moreover, the CTTN mRNA level was decreased in CRT-siRNA cells, coupled with the inactivation of STAT3. Expression of both CTTN and p-STAT3 was reduced in tumor cells following incubation with the JAK-specific inhibitor, AG490. Chromatin immunoprecipitation assay showed direct binding of p-STAT3 to the conservative STAT3-binding sequences in CTTN promoter. Furthermore, overexpression of CTTN in CRT-downregulated ESCC cells restored its motility and resistance to anoikis. This study not only reveals a role of CRT in motility promotion and anoikis resistance in ESCC cells, but also identifies CRT as an upstream regulator in the CRT-STAT3-CTTN-Akt pathway.
Subject(s)
Anoikis , Calreticulin/metabolism , Carcinoma, Squamous Cell/pathology , Cell Movement , Cortactin/metabolism , Esophageal Neoplasms/pathology , Proto-Oncogene Proteins c-akt/metabolism , STAT3 Transcription Factor/metabolism , Agar , Animals , Calreticulin/deficiency , Calreticulin/genetics , Carcinoma, Squamous Cell/genetics , Cell Line, Tumor , Cortactin/genetics , Down-Regulation , Esophageal Neoplasms/genetics , Female , Gene Expression Regulation, Neoplastic , Humans , Mice , Mice, Nude , Promoter Regions, Genetic/genetics , RNA Interference , Signal Transduction , Transcription, GeneticABSTRACT
Recent studies indicate that splicing of pre-messenger RNA and export of mRNA are normally coupled in vivo. During splicing, the conserved mRNA export factor Aly is recruited to the spliced mRNA-protein complex (mRNP), which targets the mRNA for export. At present, it is not known how Aly is recruited to the spliced mRNP. Here we show that the conserved DEAD-box helicase UAP56, which functions during spliceosome assembly, interacts directly and highly specifically with Aly. Moreover, UAP56 is present together with Aly in the spliced mRNP. Significantly, excess UAP56 is a potent dominant negative inhibitor of mRNA export. Excess UAP56 also inhibits the recruitment of Aly to the spliced mRNP. Furthermore, a mutation in Aly that blocks its interaction with UAP56 prevents recruitment of Aly to the spliced mRNP. These data suggest that the splicing factor UAP56 functions in coupling the splicing and export machineries by recruiting Aly to the spliced mRNP.
Subject(s)
Adenosine Triphosphatases/physiology , Nuclear Proteins , RNA Precursors/metabolism , RNA Splicing , RNA, Messenger/metabolism , RNA-Binding Proteins , Transcription Factors/physiology , Animals , Biological Transport , Cell Nucleus/metabolism , Cytoplasm/metabolism , HeLa Cells , Humans , Ribonucleoprotein, U2 Small Nuclear/metabolism , Spliceosomes/physiology , XenopusABSTRACT
BACKGROUND: Noncancerous cells simulating adenocarcinoma cells may interfere with the analysis of peritoneal cytology (PC) in patients with endometrial carcinoma. Immunocytochemistry (ICC) may improve the diagnosis of PC. METHODS: PC slides from 115 patients with endometrial carcinoma were reviewed. Suspicious or positive cell clusters were recovered with a cell transfer method and were subjected to ICC for MOC-31, cytokeratin 5/6, and p53. Conventional Papanicolaou staining and ICC results were compared directly on the same cells. RESULTS: By combined conventional and immunocytochemical PC (CONV-ICC-PC), cytodiagnosis was positive in 18 of 115 patients (15.7%) and suspicious in 3 of 115 patients (2.6%). According to a multivariate Cox regression analysis of patients with tumors confined to the uterus that included grade, myometrial invasion, cervical involvement, and CONV-ICC-PC, only CONV-ICC-PC was an independent prognostic factor (P < 0.05). A multivariate analysis for all of the patients studied that compared CONV-ICC-PC with staging variables revealed that only peritoneal metastasis (P < 0.0001) and lymph node metastasis (P < 0.01) were independent prognostic factors. When peritoneal metastases were excluded, CONV-ICC-PC (P < 0.01) and lymph node metastasis (P < 0.0025) were the independent prognostic factors. By cell transfer and p53 immunostaining in samples from 14 patients with malignant cells in their peritoneal washings, no deaths occurred among 5 patients with negative p53, whereas 5 of 9 patients with positive p53 died of disease at the time of data analysis. CONCLUSIONS: MOC-31 immunostaining improves the diagnosis of PC in endometrial carcinoma. Positive PC is an important prognostic factor for patients with endometrial carcinoma confined to the uterus. The p53 positive cells in PC have possible prognostic significance.
Subject(s)
Endometrial Neoplasms/pathology , Peritoneum/pathology , Adult , Aged , Aged, 80 and over , Endometrial Neoplasms/chemistry , Endometrial Neoplasms/mortality , Female , Humans , Immunohistochemistry , Keratins/analysis , Middle Aged , Prognosis , Tumor Suppressor Protein p53/analysisSubject(s)
Apoptosis , DNA/analysis , Placenta/cytology , Trophoblasts/physiology , Cell Division , Female , Humans , Pregnancy , Trophoblasts/cytologyABSTRACT
A mass outbreak of poisoning occurred in central Taiwan in 1979 due to the ingestion of rice-bran oil contaminated with polychlorinated biphenyls (PCBs). In addition to PCBs, polychlorinated dibenzofurans (PCDFs) and polychlorinated quaterphenyls (PCQs) were also found in the contaminated oil. The levels of toxic agents in the rice-oil samples collected from factory and school cafeterias and the families of the intoxicated patients were in the range of 53-99 ppm, 0.18-0.40 ppm, and 25-53 ppm for PCBs, PCDFs, and PCQs, respectively. Major components of PCBs and PCDFs in the toxic oil were separated and identified by gas chromatography (GC) and gas chromatography/mass spectrometry (GC/MS) using glass capillary columns. The most toxic PCB reported in commercial PCB preparations, 3,4,3',4'-tetrachlorobiphenyl, was found in the toxic oil as well as in one of the patients' blood and adipose tissue at an early stage of poisoning. The blood samples of 165 patients collected 9 to 18 months after the onset of poisoning contained 10 to 720 ppb of PCBs with a mean value of 38 ppb. One of the most toxic PCDFs, 2,3,4,7,8-pentachlorodibenzofuran, was retained in the liver of a deceased patient. This compound could play an important role in the etiology of PCB poisoning in Taiwan.
Subject(s)
Benzofurans/isolation & purification , Chlorobenzenes/isolation & purification , Oils/adverse effects , Polychlorinated Biphenyls/isolation & purification , Adipose Tissue/analysis , Benzofurans/blood , Chlorobenzenes/blood , Chromatography, Gas , Dibenzofurans, Polychlorinated , Food Contamination/analysis , Humans , Oils/analysis , Oryza , Polychlorinated Biphenyls/blood , Polychlorinated Biphenyls/poisoning , TaiwanABSTRACT
A mass outbreak of poisoning occurred in central Taiwan in 1979 due to the ingestion of rice-bran oil contaminated with polychlorinated biphenyls (PCBs). In addition to PCBs, polychlorinated dibenzofurans ( PCDFs ) and polychlorinated quaterphenyls ( PCQs ) were also found in the contaminated oil. The levels of toxic agents in the rice-oil samples collected from factory and school cafeterias and the families of the intoxicated patients were in the range of 53-99 ppm, 0.18-0.40 ppm, and 25-53 ppm for PCBs, PCDFs , and PCQs , respectively. Major components of PCBs and PCDFs in the toxic oil were separated and identified by gas chromatography (GC) and gas chromatography/mass spectrometry (GC/MS) using glass capillary columns. The most toxic PCB reported in commercial PCB preparations, 3,4,3'-4'-tetrachlorobiphenyl, was found in the toxic oil as well as in one of the patients' blood and adipose tissue at an early stage of poisoning. The blood samples of 165 patients collected 9 to 18 months after the onset of poisoning contained 10 to 720 ppb of PCBs with a mean value of 38 ppb. One of the most toxic PCDFs , 2,3,4,7,8-pentachlorodibenzofuran, was retained in the liver of a deceased patient. This compound could play an important role in the etiology of PCB poisoning in Taiwan.
Subject(s)
Benzofurans/poisoning , Chlorobenzenes/poisoning , Food Contamination , Foodborne Diseases/etiology , Oils/poisoning , Oryza , Polychlorinated Biphenyls/poisoning , Adipose Tissue/analysis , Benzofurans/analysis , Chlorobenzenes/analysis , Dibenzofurans, Polychlorinated , Gas Chromatography-Mass Spectrometry , Humans , Liver/analysis , Oils/analysis , Polychlorinated Biphenyls/analysis , Polychlorinated Biphenyls/blood , TaiwanSubject(s)
Chorionic Gonadotropin/blood , Postpartum Period , Pregnancy , Abortion, Induced , Female , HumansABSTRACT
In 1979, a mass outbreak of poisoning occurred in Central Taiwan due to the ingestion of rice-bran oil contaminated with polychlorinated biphenyls (PCBs). The major PCB isomers and congeners in the toxic rice oil and in the blood of PCB-poisoned patients were characterized by gas chromatography and gas chromatography/mass spectrometry using a highly efficient glass capillary column. The elimination of some major individual PCBs from blood of these patients was studied. The results indicate that tetra- and pentachlorobiphenyls with adjacent unsubstituted carbon atoms at meta-para positions are rapidly eliminated from the blood of patients, while PCBs with the same degree of chlorination but with adjacent unsubstituted carbon atoms at ortho-meta positions are eliminated more slowly. The results also indicate that most of the hexa- and heptachlorobiphenyls, with adjacent unsubstituted carbon atoms at ortho-meta positions of the biphenyl ring, are eliminated very slowly. Laboratory-animal studies have indicated that PCB excretion depends primarily on the rate of metabolism; therefore these differences in rates of elimination of PCBs should reflect the differences in their rates of metabolism.