ABSTRACT
This study extensively explored the adsorption behavior of heavy metals (Pb+2, Ni+2, Cu+2, Zn+2, and Cd+2) onto microplastics (MPs). The particle sizes of MPs ranged from 0.149 to 0.25 mm. The microplastics were generated from commercial products manufactured from both conventional (polyethylene (PE) bottle, polystyrene (PS) spoon, and polyethylene terephthalate (PET) egg carton) and biodegradable (polylactic acid (PLA) spoon, and polylactic acid (PLA) egg carton) plastics. The study also considered the influence of solution pH on the adsorption capacity of heavy metals. Regarding the adsorption potential for Cu+2, the ranking was as follows: PLA-egg (1408 µg·g-1) > PLA-spoon (735 µg·g-1) > PE-bottle (315 µg·g-1) > PET-egg (283 µg·g-1) > PS-spoon (237 µg·g-1). PLA MPs showed the highest adsorption capacity due to the lower thermal stability and higher presence of surface oxygen functional groups. Moreover, the adsorption capacities of the five metals onto PLA-spoon and PLA-egg decreased in the following order: Pb (1785 µg·g-1) > Zn (1267 µg·g-1) > Cd (748 µg·g-1) > Cu (735 µg·g-1) > Ni (722 µg·g-1), and Pb (1520 µg·g-1) > Ni (1412 µg·g-1) > Cu (1408 µg·g-1) > Zn (1118 µg·g-1) > Cd (423 µg·g-1), respectively. The SEM-EDS, FTIR and XPS results demonstrated that surface oxygen-containing functional groups play an important role during the adsorption process. This study extended its analysis to quantify the metal content of the post-adsorption MPs, revealing uneven adsorption of heavy metals onto the MPs. This implies that the diversity of commercial plastic products may result in significant variations in their ability to adsorb heavy metals, underscoring the importance of effectively managing discarded commercial plastic products.
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Acephalic spermatozoa syndrome is a rare type of teratozoospermia that severely impairs the reproductive ability of male patients, and genetic defects have been recognized as the main cause of acephalic spermatozoa syndrome. Spermatogenesis and centriole-associated 1 like (SPATC1L) is indispensable for maintaining the integrity of sperm head-to-tail connections in mice, but its roles in human sperm and early embryonic development remain largely unknown. Herein, we conducted whole-exome sequencing (WES) of 22 infertile men with acephalic spermatozoa syndrome. An in silico analysis of the candidate variants was conducted, and WES data analysis was performed using another cohort consisting of 34 patients with acephalic spermatozoa syndrome and 25 control subjects with proven fertility. We identified biallelic mutations in SPATC1L (c.910C>T:p.Arg304Cys and c.994G>T:p.Glu332X) from a patient whose sperm displayed complete acephalia. Both SPATC1L variants are rare and deleterious. SPATC1L is mainly expressed at the head-tail junction of elongating spermatids. Plasmids containing pathogenic variants decreased the level of SPATC1L in vitro. Moreover, none of the patient's four attempts at intracytoplasmic sperm injection (ICSI) resulted in a transplantable embryo, which suggests that SPATC1L defects might affect early embryonic development. In conclusion, this study provides the first identification of SPATC1L as a novel gene for human acephalic spermatozoa syndrome. Furthermore, WES might be applied for patients with acephalic spermatozoa syndrome who exhibit reiterative ICSI failures.
Subject(s)
Humans , Male , Centrioles/genetics , Homozygote , Infertility, Male/genetics , Mutation , Spermatogenesis/genetics , SpermatozoaABSTRACT
The pathogenesis of heart failure is a complex progression and associated with abnormal regulation of many signaling pathways. As a cofactor of hemoglobin, myoglobin, oxidative respiratory chain, DNA synthase and other important proteins, iron plays an indispensable role in myocardial energy metabolism. Recently, a large number of studies have shown that heart failure is related to the disorder of iron metabolism. Both iron deficiency and iron overload can lead to the development of a variety of cardiomyopathy, and even progress to heart failure. Iron metabolism could be a key target for the diagnosis, prevention and treatment of heart failure. Here, we review the basic process of iron metabolism and its mechanism in heart failure, expecting to provide new clues and evidence for the treatment of heart failure.
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PURPOSE: To assess the prevalence of fenestration and dehiscence of anterior teeth in adolescent patients with unilateral cleft lip and palate(UCLP). METHODS: Twenty-five males and seventeen females with UCLP who visited The Second Xiangya Hospital from June 2014 to September 2017 were selected. Forty-two age- and sex- matched non-cleft skeletal III patients were enrolled as the control group. Cone-beam CTï¼CBCTï¼ scans were evaluated to detect fenestration and dehiscenceï¼The data were analyzed using SPSS 22.0 software package. RESULTS: The prevalence of dehiscence of anterior teeth was 50.88% on the cleft sideï¼42.39% on the non-cleft side and 28.77% on the controls, respectively. Dehiscence of maxillary teeth was more common on cleft side than the controls (P<0.05), and dehiscence of maxillary lateral incisor was common on non-cleft side compared with the control (P<0.05). Maxillary central incisor on the cleft side showed a higher prevalence of dehiscence compared with the non-cleft side (P<0.05). Most of the dehiscence was detected on the buccal side. Fenestration of the teeth showed no significant difference among the cleft side, non-cleft side and the control (P>0.05). CONCLUSIONS: Adolescent patients with UCLP show a high prevalence of fenestration and dehiscenceï¼which deserves to be taken into consideration during clinical treatment.
Subject(s)
Cleft Lip , Cleft Palate , Adolescent , Cone-Beam Computed Tomography , Female , Humans , Male , PrevalenceABSTRACT
PURPOSE: To compare enamel surface state and operating time following debonding by using 5 different resin removal methods. METHODS: Forty healthy human premolars extracted for orthodontic purpose were selected and then divided into 5 groups randomly based on different resin-removal methods, 8 in each group. After regular bonding brackets and debonding, tungsten carbide bur (group A), tungsten carbide bur+silicon particles(group B), corundum bur(group C), corundum bur+silicon particles (group D) and silicon particles (group E) were used respectively to deal with the enamel surfaces. The operating time(T) of each group was recorded. The samples were observed under scanning electron microscope (SEM), and enamel damage index(EDI) was recorded. The scores of EDI were analysed using SPSS 20.0 software package . RESULTS: The order of EDI was group C>group D>group A >group B>group E, significant difference was found between 5 groups (P<0.05). The order of operating time was group E>group D>group B>group A>group C, significant difference was found between 5 groups (P<0.05). CONCLUSIONS: Using tungsten carbide bur followed by silicon particles to remove residual resin on enamel surface causes less damage to the enamel surface, and requires reasonable time.
Subject(s)
Dental Bonding , Dental Debonding , Orthodontic Brackets , Acrylic Resins , Bicuspid , Dental Enamel , Humans , Microscopy, Electron, Scanning , Random Allocation , Surface PropertiesABSTRACT
The aim of the present study was to investigate the effect of salidroside (Sal) on inflammatory activation induced by lipopolysaccharide (LPS) in the co-culture of rat alveolar macrophages (AM) NR 8383 and type II alveolar epithelial cells (AEC II) RLE-6TN. CCK-8 colorimetric method was used to detect cell proliferation percentage. The enzyme-linked immunosorbent assay (ELISA) was used to determine the content of tumor necrosis factor alpha (TNF-α), macrophage inflammatory protein-2 (MIP-2) and interleukin-10 (IL-10) in the supernatant. Western blot was used to examine the expression levels of phosphorylated AKT (p-AKT) and total AKT protein. The results showed that pretreatment of RLE-6TN cells or co-culture of RLE-6TN and NR 8383 cells with 32 and 128 µg/mL Sal for 1 h, followed by continuous culture for 24 h, significantly increased the cell proliferation (P < 0.05). Compared with control group, 32 and 128 µg/mL Sal pretreatment significantly increased the ratio of p-AKT/AKT in RLE-6TN cells (P < 0.05). Pretreatment of 32 µg/mL Sal not only inhibited the secretion of TNF-α and MIP-2 by NR 8383 cells induced by LPS (P < 0.05), but also enhanced the inhibitory effect of RLE-6TN and NR 8383 cells co-culture on the secretion of TNF-α and MIP-2 by NR 8383 cells induced by LPS (P < 0.05). In addition, 32 µg/mL Sal pretreatment promoted LPS-induced IL-10 secretion by NR 8383 cells (P < 0.05), and enhanced the promoting effect of co-culture of RLE-6TN and NR 8383 cells on the IL-10 secretion by LPS-induced NR 8383 cells (P < 0.05). In conclusion, Sal may directly inhibit LPS-induced inflammatory activation of AM (NR 8383), promote the proliferation of AEC II (RLE-6TN) through PI3K/AKT signaling pathway, and enhance the regulatory effect of AEC II on LPS-induced inflammatory activation of AM.
Subject(s)
Animals , Rats , Alveolar Epithelial Cells , Metabolism , Cell Line , Chemokine CXCL2 , Metabolism , Coculture Techniques , Glucosides , Pharmacology , Interleukin-10 , Metabolism , Lipopolysaccharides , Macrophages, Alveolar , Metabolism , Phenols , Pharmacology , Phosphatidylinositol 3-Kinases , Metabolism , Proto-Oncogene Proteins c-akt , Metabolism , Signal Transduction , Tumor Necrosis Factor-alpha , MetabolismABSTRACT
Objectives: To analyze and understand the risk factors related to HIV new infections among men who have sex with men (MSM). Methods: A longitudinal observational study among MSM was conducted to collect information on HIV related behaviors and sero-conversion. Univariate and multivariate generalized estimating equations (GEE) were used to discuss the risk factors for HIV new infection. Results: A total number of 4 305 MSM were followed during 2013-2015. Among those self-reported MSM who are seeking partners on the Interner tended to have higher proportion on receptive anal intercourse and consistent condom use during anal intercourse than the subgroups seeking their partners in gay bars or bathrooms. HIV incidence among followed MSM during the study period appeared as 4.3/100 person years, with adjusted RR (aRR) of HIV infection for receptive anal intercourse as group 2.20 (95% CI: 1.49-3.24) times than that of insertion anal intercourse group. Those who used rush-poppers (aRR=1.55, 95% CI: 1.10-2.17), unprotected anal intercourse (aRR=2.24, 95%CI: 1.62-3.08), and those with syphilis infection (aRR=2.95, 95%CI: 2.00-4.35) were also risk factors for HIV new infections. After controlling other factors, the relationship between the ways of seeking partners and HIV new infection was not statistical significant. Conclusion: Risk factors for HIV new infection among MSM appeared complex and interactive, suggesting that further studies are needed to generate tailored strategies for the prevention of HIV epidemic among MSM population.
Subject(s)
Adult , Humans , Male , Cohort Studies , HIV Infections/diagnosis , Homosexuality, Male , Incidence , Risk Factors , Risk-Taking , Sexual Behavior , Sexual Partners , Sexual and Gender Minorities , Surveys and Questionnaires , Unsafe SexABSTRACT
The flooding-drying cycles in the water-level fluctuation zone (WLFZ) of the Three Gorges Reservoir (TGR) result in an abundance of plants that are submerged and decomposed to release nutrients. This has the potential to lead to water quality deterioration of the TGR. Cynodon dactylon (Linn.) Pers., one of the typical plants in the WLFZ, was collected and inundated in the laboratory under different conditions under temperatures of 15â and 25â, a pH 5.0 to 9.0, and under light and dark conditions, respectively, to measure the decomposition rate and nitrogen and phosphorus release mechanisms. The dry weight lost with the contents of nitrogen and phosphorus was found to decrease after Cynodon dactylon (Linn.) Pers. was soaked for decomposition. The amount released of total nitrogen (TN) and total phosphorus (TP) at 25â for 200-days inundation was(2.66±0.29)g·kg-1 or(3.76±0.04)g·kg-1, respectively and at 15â was(0.79±0.03) g·kg-1 or (1.40±0.02) g·kg-1, respectively. When the pH of the water submerging the grass was 5.0, 7.0 and 9.0, the amount of TN released was (3.76±0.08) g·kg-1, (2.66±0.29) g·kg-1, and (2.55±0.12) g·kg-1, respectively while the amount of TP released was (1.53±0.04) g·kg-1, (0.79±0.03) g·kg-1, and (1.70±0.07) g·kg-1, respectively. The TN and TP released was (3.87±0.14) g·kg-1 and (1.78±0.08) g·kg-1 under dark condition. The lower the temperature, the higher the amount of TN and TP will be released for inundation from Cynodon dactylon (Linn.) Pers. When the overlying water is acidic or alkaline, more TN and TP is released. Dark conditions are beneficial to nitrogen and phosphorus release into the overlying water. Thus, the water environment changes in each flooding season in winter. Sewage discharge will also accelerate the nutrients released from soaked plants through their decomposition in the WLFZ, and then will aggravate the deterioration of water quality in TGR.
Subject(s)
Cynodon/metabolism , Fresh Water/analysis , Nitrogen/metabolism , Phosphorus/metabolism , Water Pollutants, Chemical/metabolism , China , Floods , Water QualityABSTRACT
To study the protective effect and mechanism of synthetic salidroside on acute lung injury (ALI) induced by lipopolysaccharide (LPS), male Sprague-Dawley (SD) rats were randomly divided into saline control group, 3 mg/kg LPS model group, different doses of salidroside groups (5, 20 and 80 mg/kg), and 5 mg/kg dexamethasone group. Intratracheal LPS instillation was used to establish the ALI model 0.5 h after intraperitoneal injection of salidroside or dexamethasone, and the rats were sacrificed 6 h later. Lung wet/dry weight ratio (W/D) was calculated. Lung tissue pathology and lung injury score (LIS) were observed and evaluated through hematoxylin and eosin (HE) staining. The centrifugal sediment of bronchoalveolar lavage fluid (BALF) was used to count the polymorphonuclear leukocyte (PMN) number by Wright's staining, and the centrifugal supernatant of BALF was used to determine the contents of protein and inflammatory factors (TNF-α, IL-1β and IL-6). The contents of myeloperoxidase (MPO) and malondialdehyde (MDA) in lung tissue were determined. Western blot was used to detect the expression levels of phosphorylated and total nuclear factor kappa B (NF-κB)/p65 protein in lung tissue. The results showed that, compared with LPS group, the intervention of synthetic salidroside alleviated the pathological damage in lung tissue, decreased the LIS and lung W/D ratio (P < 0.05), reduced the PMN number, the contents of protein and inflammatory factors in BALF (P < 0.05), reduced the contents of MPO and MDA in lung tissue (P < 0.05), and inhibited the expression of p-NF-κB in lung tissue (P < 0.05). The results suggest that synthetic salidroside has a protective effect on ALI induced by LPS, and its mechanism is related to inhibiting the phosphorylation of NF-κB and reducing the aggregation of PMN in the lung.
Subject(s)
Animals , Male , Rats , Acute Lung Injury , Drug Therapy , Bronchoalveolar Lavage Fluid , Dexamethasone , Pharmacology , Glucosides , Pharmacology , Interleukin-1beta , Metabolism , Interleukin-6 , Metabolism , Lipopolysaccharides , Lung , Pathology , Malondialdehyde , Metabolism , NF-kappa B , Metabolism , Neutrophils , Cell Biology , Peroxidase , Metabolism , Phenols , Pharmacology , Phosphorylation , Random Allocation , Rats, Sprague-Dawley , Tumor Necrosis Factor-alpha , MetabolismABSTRACT
Our earlier findings indicate that the long non-coding RNA MALAT1 promotes colorectal cancer (CRC) cell proliferation, invasion and metastasis in vitro and in vivo by increasing expression of AKAP-9. In the present study, we investigated the molecular mechanism by which MALAT1 enhances AKAP9 expression in CRC SW480 cells. We found that MALAT1 interacts with both SRPK1 and SRSF1. MALAT1 increases AKAP-9 expression by promoting SRPK1-catalyzed SRSF1 phosphorylation. Following MALAT1 knockdown, overexpression of SRPK1 was sufficient to restore SRSF1 phosphorylation and AKAP-9 expression to a level that promoted cell proliferation, invasion and migration in vitro. Conversely, SRPK1 knockdown after overexpression of MALAT1 in SW480 cells diminished SRSF1 phosphorylation and AKAP-9 expression and suppressed cell proliferation, invasion and migration in vitro. These findings suggest MALAT1 increases AKAP-9 expression by promoting SRPK1-catalyzed SRSF1 phosphorylation in CRC cells. These results reveal a novel molecular mechanism by which MALAT1 regulates AKAP-9 expression in CRC cells.
Subject(s)
A Kinase Anchor Proteins/metabolism , Colorectal Neoplasms/metabolism , Cytoskeletal Proteins/metabolism , Protein Serine-Threonine Kinases/metabolism , RNA, Long Noncoding/metabolism , Serine-Arginine Splicing Factors/metabolism , Cell Line, Tumor , Cell Proliferation/physiology , Colorectal Neoplasms/genetics , Colorectal Neoplasms/pathology , Humans , Phosphorylation , RNA, Long Noncoding/genetics , Serine-Arginine Splicing Factors/genetics , TransfectionABSTRACT
N-myc downstream-regulated gene 1 (NDRG1) has been implicated in tumorigenesis and metastasis in different cancers. However, its role in nasopharyngeal carcinoma remains unknown. We found that NDRG1 expression level was high in nasopharyngeal cancer 5-8F cells but low in 5-8F-LN cells with lymphatic metastasis potential. Knockdown of NDRG1 by shRNA promoted 5-8F cell proliferation, migration, and invasion in vitro and its tumorigenesis in vivo. Moreover, NDRG1 deficiency induced an epithelial-mesenchymal transition (EMT) of 5-8F cells as shown by an attenuation of E-cadherin and an induction of N-cadherin and vimentin expression. NDRG1 knockdown also enhanced Smad2 expression and phosphorylation. Smad2 signaling was attenuated in 5-8F cells but was significantly activated in 5-8F-LN cells. Knockdown of Smad2 restored E-cadherin but attenuated N-cadherin expression in NDRG1-deficient 5-8F cells, suggesting a reduction of EMT. Consistently, blockade of Smad2 in 5-8F-LN cells increased E-cadherin while diminishing N-cadherin and vimentin expression. These data indicate that Smad2 mediates the NDRG1 deficiency-induced EMT of 5-8F cells. In tumors derived from NDRG1-deficient 5-8F cells, E-cadherin expression was inhibited while vimentin and Smad2 were increased in a large number of cancer cells. Most importantly, NDRG1 expression was attenuated in human nasopharyngeal carcinoma tissues, resulted in a lower survival rate in patients. The NDRG1 was further decreased in the detached nasopharyngeal cancer cells, which was associated with a further reduced survival rate in patients with lymphatic metastasis. Taken together, these results demonstrated that NDRG1 prevents nasopharyngeal tumorigenesis and metastasis via inhibiting Smad2-mediated EMT of nasopharyngeal cells.
ABSTRACT
OBJECTIVE: This study aims to explore the function of Integrin-ß/FAK in the mechanical signal transduction and the connection with downstream ERK signal pathways. METHODS: Human osteosarcoma MG63 cell lines were used in this study. The effects of mechanical strain on the Integrin-ß1 expression, FAK and ERK signal pathway in Human osteosarcoma MG63 cells were detected using RT-PCR and Western-blotting methods. The localization of FAK in Human osteosarcoma MG63 cells were determined using immunofluorescent method. The interaction between Integrin-ß1 and FAK were detected by using co-immunoprecipitation method. RESULTS: The expression of Integrin-ß1 shows a notable bimodel distribution, mechanical strain stimulation can promote Integrin-ß1 expression and the phosphorylation of FAK and ERK, mechanical strain activated FAK and ERK mediated by Integrin-ß1. CONCLUSION: Integrin-ß1 may play an important role in osteoblast proliferation differentiation process, it might feel external strain stimulation through ECM composition and makes FAK phosphated through the interaction with FAK, thus causing a series of activation of signal molecules. Finally it reduces MAPK (ERK) activation and cellular responses to finish mechanical signal transduction.
Subject(s)
Focal Adhesion Kinase 1/metabolism , Integrin beta1/metabolism , MAP Kinase Signaling System , Cell Differentiation , Cell Line, Tumor , Cell Proliferation , Focal Adhesion Kinase 1/genetics , Humans , Integrin beta1/genetics , PhosphorylationABSTRACT
<p><b>OBJECTIVE</b>To study the possible clonal origin of neuroendocrine cells in colorectal adenocarcinoma.</p><p><b>METHODS</b>Twenty-six microsatellite loci were screened using laser capture microdissection, DNA extraction and whole genome amplification. Microsatellite instability (MSI) and loss of heterozygosity (LOH) in adenocarcinoma cells and neuroendocrine cells amongst 30 cases of colorectal carcinoma with neuroendocrine differentiation were detected using polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP)-silver staining. The mutation status of p53 was evaluated by PCR-sequencing. The clonal origin of neuroendocrine cells in colorectal adenocarcinoma was determined.</p><p><b>RESULTS</b>Amongst the 30 cases studied, the prevalence of MSI was 16.9% while that of LOH was 8.5%. The rate showed no statistically significant difference between adenocarcinoma cells and neuroendocrine cells. In 6 cases, the microsatellite alteration was entirely consistent. In 23 cases, the rate of microsatellite alteration consistency was greater than that of inconsistency. In 1 case, the consistency and inconsistency rates were identical. There was statistically significant difference between consistency and inconsistency of microsatellite alteration. The prevalence of p53 mutation was 16.7% which was the same for both adenocarcinoma cells and neuroendocrine cells.</p><p><b>CONCLUSIONS</b>Adenocarcinoma cells and neuroendocrine cells in colorectal adenocarcinoma with neuroendocrine differentiation have similar biologic changes. It is likely that they are of identical origin.</p>
Subject(s)
Humans , Adenocarcinoma , Genetics , Pathology , Colorectal Neoplasms , Genetics , Pathology , DNA Mutational Analysis , Laser Capture Microdissection , Loss of Heterozygosity , Microsatellite Instability , Neuroendocrine Cells , Pathology , Tumor Suppressor Protein p53 , GeneticsABSTRACT
The water-coordinated Ni(2+) cation in the title compound, [Ni(C(10)H(5)N(3)O(4))(H(2)O)](n), assumes an octa-hedral NiN(3)O(3) coord-ination mode and is N,O-chelated by two deprotonated 2-(pyridin-4-yl)-1H-imidazole-4,5-dicarb-oxy-lic acid (HPyImDC(2-)) ligands, forming a layer structure extending in the bc plane. The chains are arranged along the b-axis direction, forming a layer structure extending in the bc plane. O-Hâ¯O hydrogen bonding between the layers results in the formation of a three-dimensional supra-molecular framework. The structure is isotypic with the Zn analogue [Li et al. (2009). Cryst. Growth Des.6, 3423-3431].
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To explore the possible role for connective tissue growth factor (CTGF) during tooth movement, we evaluated CTGF gene and protein expression in MG-63 cells subjected to cyclic stretch. Cyclic stretch caused a time-dependent increase in CTGF mRNA and protein levels.Inhibition of p38 MAP kinase or ERK activation did not affect cyclic stretch-induced CTGF expression. Specific inhibitors of PI3K suppressed stretch -induced CTGF expression in a time-dependent manner. cyclic stretch activated JNK and ERK, but not p38 MAP kinase in osteoblast-like cells. PI3K inhibitors suppressed cyclic stretch-induced JNK, but not p38 MAP kinase activation. Finally, SP600125, a Specific Inhibitor of JNK, suppressed stretch -induced CTGF Expression. These results suggest that stretch-induced CTGF expression is mediated through the PI3K-JNK -dependent pathway, not by p38 MAP kinase and ERK pathways.
Subject(s)
Connective Tissue Growth Factor/metabolism , Osteoblasts/metabolism , Signal Transduction , Stress, Mechanical , Anthracenes/pharmacology , Cell Line , Chromones/pharmacology , Connective Tissue Growth Factor/genetics , Extracellular Signal-Regulated MAP Kinases/antagonists & inhibitors , Extracellular Signal-Regulated MAP Kinases/metabolism , Flavonoids/pharmacology , Humans , JNK Mitogen-Activated Protein Kinases/antagonists & inhibitors , JNK Mitogen-Activated Protein Kinases/metabolism , Morpholines/pharmacology , Osteoblasts/drug effects , Phosphatidylinositol 3-Kinases/metabolism , Phosphoinositide-3 Kinase Inhibitors , p38 Mitogen-Activated Protein Kinases/metabolismABSTRACT
OBJECTIVE: To explore certain principle of how osteoprotegerin (OPG) and osteoprotegerin ligand (OPGL) take part in the periodontal tissues remodeling under the combined influence of inflammation and orthodontic force. METHODS: The positive signals of OPG and OPGL mRNA were measured with in situ hybridzation after orthodontic tooth movement in the experimental periodontitis groups and control ones. RESULTS: The OPG and OPGL mRNA expression intensity in the experimental group showed difference from control. All their optical density index reached a peak in day 2, respectively. CONCLUSION: OPG and OPGL play important roles in the periodontal reconstruction induced by inflammation irritation and orthodontic force, and complex interaction could exist between the two factors.
Subject(s)
Osteoprotegerin , RANK Ligand , Humans , Periodontitis , RNA, Messenger , Tooth Movement TechniquesABSTRACT
OBJECTIVE: To study the integrin beta3 mRNA changes after orthodontic treatment on normal teeth and periodontitis teeth in rats. METHODS: 96 adult SD rats of 10 weeks old were randomly divided into normal tooth move-ment group and periodontitis tooth movement group. The rats in the two groups were sacrificed after 0 d, 12 h, 1 d, 3 d, 5 d and 7 d of tooth movement. The alveolar specimens were prepared. The integrin beta3 mRNA were detected using in situ hybridization in the specimens. The OD index of positively stained osteoclasts for integrin beta3 mRNA after orthodontic tooth movement in the two groups were measured and compared. RESULTS: There were weak positive signals on the cytoplasm of osteoclasts in periodontum in both groups after 12 hours and 3 days force activation. No positive signals were detected in the rest samples. There was no difference in the OD of positive stained osteoclasts between normal and periodontitis groups. Strong expressions were present on cells with one or two nuclei in the alveolar marrow. CONCLUSION: It is suggested that integrin beta3 mRNA is related with osteoclasts maturation and migration in orthodontic tooth movement.
Subject(s)
RNA, Messenger , Rats, Sprague-Dawley , Animals , In Situ Hybridization , Integrins , Osteoclasts , Periodontitis , Rats , Tooth Movement TechniquesABSTRACT
<p><b>OBJECTIVE</b>To construct the prokaryotic and eukaryotic expression vectors pET32/E5 and pcDNA3.1/E5 for transformation into E. coli BL21 and NIH(3)T(3) cells respectively to observe the expression of human papillomavirus type 16 E5 protein (HPV16 E5).</p><p><b>METHODS</b>HPV16 E5 gene was amplified by PCR from clinical isolates of HPV 16 and inserted into the plasmid pET32a(+) followed by digestion with BamH I and Hind III. The recombinant plasmid pET32/E5 was transformed into E. coli JM109 and selected with ampicillin. The positive clones containing the recombinant plasmid pET32/E5 were verified by restriction endonucleases BamH I and Xho and sequence analysis. The expression of HPV16 E5-TRX fusion protein in E. coli BL21(ED3) was identified by SDS-PAGE and Western blotting. The digestion product of BamH I and Xho was purified and inserted into the eukaryotic expression vector pcDNA3.1(+) to construct pcDNA3.1/E5, which was identified by sequencing and transfected into NIH3T3 cells. The NIH(3)T(3) cells with stable expression of HPV16 E5 were selected by G418 and confirmed by RT-PCR.</p><p><b>RESULTS</b>The pET32/E5 and pcDNA3.1/E5 vectors were constructed successfully. E.coli BL21(DE3) transformed by the recombinant plasmid pET32/E5 expressed HPV16 E5-TRX fusion protein efficiently. In the presence of 1 mmol/L IPTG at 28 degrees C, HPV16 E5-TRX recombinant protein accounted for about 10% of the total bacterial proteins. NIH3T3 cells stably expressing HPV16 E5 were harvested by selection with 250 g/ml of G418. HPV16 E5 gene from pcDNA3.1/E5-transfected NIH(3)T(3) cells was amplified by RT-PCR, and sequence analysis demonstrated the acquisition of the full-length gene fragment.</p><p><b>CONCLUSIONS</b>The prokaryotic and eukaryotic vectors for the HPV16 E5 gene have been successfully constructed. The acquisition of E .coli and NIH(3)T(3) cells with stable expression of HPV16 E5 protein may facilitate subsequent research of the biological properties and the transformation mechanism of HPV16 E5 protein on specific cells.</p>
Subject(s)
Animals , Humans , Mice , 3T3 Cells , Escherichia coli , Genetics , Metabolism , Eukaryotic Cells , Metabolism , Genetic Vectors , Oncogene Proteins, Viral , Genetics , Papillomaviridae , Genetics , Papillomavirus Infections , Virology , Plasmids , Genetics , Prokaryotic Cells , Metabolism , Recombinant Fusion Proteins , GeneticsABSTRACT
OBJECTIVE: To study the integrin beta1 mRNA changes after orthodontic tooth movement in normal teeth and periodontitis teeth of rats. METHODS: The OD of positively stained osteoclasts for integrin beta1 mRNA using in situ hybridzation was detected after orthodontic tooth movement in normal teeth and periodontitis teeth groups. RESULTS: Integrin beta1 mRNA expression were detected on all osteoclasts in tooth movement samples of normal and periodontitis teeth. There were stronger positive signals after given orthodontic force in both of the two groups. But no differences were found after 0.5, 1, 2, 3, 5, 7, 10 days since orthodontic tooth movement. The integrin beta1 mRNA signals in normal tooth movement group were not different from that in periodontitis group. CONCLUSION: The integrin beta1 of osteoclasts may play a role in the stability and remodeling of periodontal ligament in orthodontic tooth movement. There were no difference in the OD of integrin beta1 mRNA staining in orthodontic tooth movement between normal teeth group and periodontitis teeth group.
Subject(s)
Integrin beta1/metabolism , Periodontitis/physiopathology , Tooth Movement Techniques , Animals , Osteoclasts , Periodontal Ligament , RNA, Messenger/metabolism , RatsABSTRACT
OBJECTIVE: To study the bracket placement and arch wire bending based on ethnic differences and individual differences of normal occlusion. METHODS: The prominence, tip, torque, upper first molar offset of crown and arch form between Chinese and Caucasian normal occlusion were compared. RESULTS: The results showed the ethnic differences of prominence, tip, torque, upper first molar offset of crown and arch form between Chinese and Caucasian normal occlusion. The placement of bracket was influenced by the crown morphology. CONCLUSION: The adjustments of the bracket placement and arch wire bending with Edgewise and pre-adjusted appliance are necessary to adapt to ethnic difference and individual difference.