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1.
Chemphyschem ; 21(20): 2249-2253, 2020 10 16.
Article in English | MEDLINE | ID: mdl-32869462

ABSTRACT

The thermodynamically unstable, colourless closed-ring isomer of spiropyran can be stabilized in water by the anti-configurational isomer of amide naphthotube. The influence of the binding on the thermodynamics and kinetics of spiropyran have been studied. The complex was further used to prepare a test paper that allows naked-eye detection of toxic paraoxon.

2.
Anal Biochem ; 544: 98-107, 2018 03 01.
Article in English | MEDLINE | ID: mdl-29305096

ABSTRACT

With the emergence of multi- and extensive-drug (MDR/XDR) resistant Mycobacterium tuberculosis (M. tb), tuberculosis (TB) persists as one of the world's leading causes of death. Recently, isothermal DNA amplification methods received much attention due to their ease of translation onto portable point-of-care (POC) devices for TB diagnosis. In this study, we aimed to devise a simple yet robust detection method for M. tb. Amongst the numerous up-and-coming isothermal techniques, Recombinase Polymerase Amplification (RPA) was chosen for a real-time detection of TB with or without MDR. In our platform, real-time RPA (RT-RPA) was integrated on a lab-on-a-disc (LOAD) with on-board power to maintain temperature for DNA amplification. Sputa collected from healthy volunteers were spiked with respective target M. tb samples for testing. A limit of detection of 102 colony-forming unit per millilitre in 15 min was achieved, making early detection and differentiation of M. tb strains highly feasible in extreme POC settings. Our RT-RPA LOAD platform has also been successfully applied in the differentiation of MDR-TB from H37Ra, an attenuated TB strain. In summary, a quantitative RT-RPA on LOAD assay with a high level of sensitivity was developed as a foundation for further developments in medical bedside and POC diagnostics.


Subject(s)
Automation , Lab-On-A-Chip Devices , Mycobacterium tuberculosis/genetics , Nucleic Acid Amplification Techniques , Polymerase Chain Reaction , Tuberculosis, Multidrug-Resistant/genetics , Healthy Volunteers , Humans , Point-of-Care Testing , Time Factors
3.
Postgrad Med J ; 84(995): 498-501, 2008 Sep.
Article in English | MEDLINE | ID: mdl-18940951

ABSTRACT

BACKGROUND: It has been recognised that genetic or hereditary factors may contribute to the aetiology of adolescent idiopathic scoliosis (AIS). Recently, two linkage analyses have identified 19p13.3 as the candidate region for AIS. The dipeptidyl peptidase 9 (DPP9) gene is located on chromosome 19p13.3. OBJECTIVE: To investigate whether DPP9 gene polymorphisms are associated with the occurrence or curve severity of AIS. METHODS: 571 girls with AIS and 236 normal controls were recruited. Using the Chinese data from the HapMap project, a set of tagging single-nucleotide polymorphisms (tagSNPs) were defined for the DPP9 gene. Five SNPs were genotyped by PCR restriction fragment length polymorphism. Statistical analysis of genotype frequencies between cases and controls was performed by the chi2 test. One-way analysis of variance was used to compare mean maximum Cobb angles with different genotypes in case-only analysis. RESULTS: Genotype frequencies were comparable between cases and controls for all five polymorphisms (p>0.05). The mean maximum Cobb angles of different genotypes were similar to each other for all five polymorphisms. CONCLUSIONS: The DPP9 gene is not associated with the occurrence or curve severity of AIS. It is neither a disease-predisposition nor a disease-modifying gene of AIS.


Subject(s)
Dipeptidyl-Peptidases and Tripeptidyl-Peptidases/genetics , Scoliosis/genetics , Adolescent , Child , Female , Genetic Linkage/genetics , Humans , Polymorphism, Genetic/genetics
4.
Bone ; 40(3): 685-92, 2007 Mar.
Article in English | MEDLINE | ID: mdl-17188589

ABSTRACT

OBJECTIVES: This study tested the effect of Epimedium-derived phytoestrogen (PE) on preventing steroid-associated osteonecrosis (ON) in rabbit model. METHODS: Thirty 28-week-old male New-Zealand white rabbits were divided into control group (CON; n=14) and PE group (PE; n=16; 5 mg/kg body weight/day) after receiving an established inductive protocol for inducing steroid-associated ON. Before and after inductive protocol, Dynamic-MRI was employed on bilateral femora for local intra-osseous perfusion, blood samples were examined for coagulation, fibrinolysis and lipid-transportation, and marrow samples were quantified for adipogenesis-gene mRNA expression. Six weeks later, bilateral femora were dissected for Micro-CT-based micro-angiography, and then ON lesion, intravascular thrombosis and extravascular fat-cell-size were examined histopathologically. RESULTS: The incidence of ON in the PE group (31%) was significantly lower than that in the CON group (93%). Compared to the CON group, local intra-osseous perfusion was maintained in the PE group. Blocked trunk vessels were seldom found in micro-angiography of the PE-treated rabbits. Thrombosis incidence and fat-cell-size were both significantly lower in the PE group than those in the CON group. During the early period after induction, indicator of coagulation, fibrinolysis, lipid-transportation and adipogenesis-gene expression were found with significantly changing pattern in the PE group compared to the CON group. CONCLUSION: PE was able to exert beneficial effect on preventing steroid-associated ON in rabbits with inhibition of both thrombosis and lipid deposition.


Subject(s)
Epimedium/chemistry , Osteonecrosis/prevention & control , Phytoestrogens/therapeutic use , Phytotherapy , Plant Preparations/therapeutic use , Adrenal Cortex Hormones/adverse effects , Animals , Femur/blood supply , Femur/drug effects , Femur/pathology , Lipid Metabolism/drug effects , Magnetic Resonance Imaging , Male , Osteonecrosis/chemically induced , Rabbits , Reverse Transcriptase Polymerase Chain Reaction , Thrombosis/prevention & control
5.
Article in English | MEDLINE | ID: mdl-17108395

ABSTRACT

UNLABELLED: Many studies have demonstrated the role of melatonin in the etiology of AIS. Previous studies have shown that there is no evidence of mutations in the melatonin receptor 1A gene in AIS patients. In this study, we have examined the role of melatonin receptor 1B in predisposition for AIS. Using haplotype block tagging technique, a set of tagging SNPs were defined for MTNR1B from the Han Chinese data of the International HapMap project. The association between the tagging of single nucleotide polymorphisms (tSNPs) in MTNR1B region and the occurrence of AIS was studied. METHOD: 473 AIS girls and 311 normal controls were recruited. The age range of the patients was between 10 and 18 years old. The maximum Cobb was recorded at latest follow-up in AIS patients. Three of five tSNPs were studied; they were all located within the coding region of the MTNR1B gene. RESULTS: There was no significant difference in the genotype or allelic frequencies (AF) of the 3 tSNPs between AIS and controls. In a case-only analysis, no difference in curve severity in AIS patients was found among patients with different genotypes (by one-way ANOVA). DISCUSSION: The 3 tSNPs showed no association with either the occurrence of AIS or the maximum Cobb angle within AIS girls. Further analysis of the remaining tSNPs within the regulatory region of the MTNR1B gene and other related genes in the melatonin signaling pathway may provide further information on the role of the melatonin in AIS girls.


Subject(s)
Genetic Predisposition to Disease , Receptors, Melatonin/genetics , Scoliosis/genetics , Adolescent , Child , China , Female , Humans , Male , Polymorphism, Genetic , Scoliosis/etiology , Sequence Analysis, DNA
6.
Stud Health Technol Inform ; 123: 18-24, 2006.
Article in English | MEDLINE | ID: mdl-17108398

ABSTRACT

UNLABELLED: IGF-I has a pivotal role in bone growth and could be one of the putative disease-modifier genes in AIS. Two SNPs in IGF-I gene promoter region were studied for any association with occurrence of AIS and for their effect on the curve severity among AIS. METHODS: 506 AIS girls (Cobb>20 degrees) and 227 age-matched Chinese girls were recruited. The spine (L2-L4) and hip BMD of the subjects were measured by DXA. A subgroup of AIS patients (N=340) who were followed-up to skeletal maturity and the maximum Cobb's angle was recorded. Two SNPs were genotyped by PCR-RFLP (rs5742612 and rs2288377). The chi-square test and one-way ANOVA were used to test the association between genotypes and quantitative parameters, respectively. RESULTS: No association was between the genotypes and the occurrence of AIS and the BMD of the spine and hip. The allelic frequency of T allele was 0.69 in AIS and control. However, the Cobb's angle was higher in patients with the homozygous T allele (Mean Cobb's angle: 38.1 degrees in TT vs 35.9 degrees in TC vs 33.2 degrees in CC group; p=0.04). DISCUSSION: Interestingly, IGF-I polymorphism affects the curve severity of AIS though it was not associated with onset of AIS per se. It indicates that IGF-I may be a disease modifying gene. The importance of IGF-I in skeletal growth makes it a good candidate gene which would play a role in the documented association of rapid growth with curve progression in AIS.


Subject(s)
Bone Diseases, Metabolic , Insulin-Like Growth Factor I/genetics , Scoliosis/genetics , Scoliosis/physiopathology , Adolescent , Child , Female , Humans
7.
Stud Health Technol Inform ; 123: 27-33, 2006.
Article in English | MEDLINE | ID: mdl-17108399

ABSTRACT

With the use of multiplanar reformat Magnetic Resonance imaging, AIS patients were found to have significantly reduced pedicle widths on concavity. Pattern of vertebral asymmetry was also exaggerated with smaller pedicle width, length and area on concavity. The cord appeared more roundish and was deviated to the concavity at apical vertebra in AIS. A tethering force might therefore be present on the cord along the transverse axis in AIS, accounted by the relatively fixed position of the exit nerve roots and deviation of the cord from the exit foraminae of the corresponding vertebra.


Subject(s)
Lumbar Vertebrae/innervation , Magnetic Resonance Imaging/methods , Scoliosis/physiopathology , Adolescent , Child , Female , Hong Kong , Humans , Lumbar Vertebrae/anatomy & histology , Lumbar Vertebrae/physiopathology
8.
Stud Health Technol Inform ; 123: 47-51, 2006.
Article in English | MEDLINE | ID: mdl-17108402

ABSTRACT

The aim of this study was to monitor BMD changes occurring during periods of rapid growth and to investigate whether osteopenia was a persistent phenomenon in skeletally matured AIS girls. 196 AIS Chinese girls and 122 healthy controls, aged 11-16, were follow-up for 3.5 years. Bilateral femoral neck bone mineral density (BMD) and volumetric BMD (vBMD) of the distal tibia were obtained by dual energy X-ray absorptiometry (DXA) and peripheral quantitative computed tomography (pQCT). Osteopenia was defined if the age-adjusted BMD was below or equal to -1 standard deviation (SD). The average age at the final follow-up was 16.8 years old. The median initial Cobb angle for this group of patients was 26 degrees. The prevalence of osteopenia at the baseline measurement was 35.9%. Longitudinal BMD results demonstrated that 86.0% of osteopenic AIS girls had persistently low BMD at the time of skeletal maturity (age of 16). vBMD of distal tibia of AIS was significantly lower than that of the controls throughout the age of 13 to 17 during the period of rapid growth. In addition, there were also significant differences in vBMD among AIS (moderate and severe group) and the controls by one-way ANOVA (p<0.05). The present study for the first time revealed that over 86% of osteopenic AIS patients had persistently low BMD, at both distal tibia and femoral neck regions, at the time of skeletal maturity. Early detection and treatment of AIS-related osteopenia might help in maximizing peak bone mass during peripubertal growth that thereby minimizing risks of developing osteoporotic fractures later in life.


Subject(s)
Bone Density/physiology , Bone Diseases, Metabolic/physiopathology , Monitoring, Physiologic , Scoliosis/physiopathology , Adolescent , Child , Female , Hong Kong , Humans
9.
Stud Health Technol Inform ; 123: 599-604, 2006.
Article in English | MEDLINE | ID: mdl-17108495

ABSTRACT

UNLABELLED: A lowered bone mineral density (BMD) was observed in trabecular bone in a significant number of AIS patients. The present study aims at investigating whether lower BMD is a systemic phenomenon, which would also be present in the cortical component of long bone. SUBJECTS AND METHODS: 78 AIS girls (age:15-18y.o) with either moderate (Cobb: 20-40) or severe (Cobb: >40) curve and 44 age-matched controls were recruited. The BMD of the distal region and the mid-shaft of radius were measured with a multi-layer peripheral quantitative computed tomography (pQCT). The trabecular bone and cortical bone BMD and the morphology of mid-shaft were compared. RESULTS: Both trabecular and cortical BMD in severe AIS group was significantly lower than the control by 8.7% and 1.7% (p<0.05 for both), respectively. However, the cortical bone area of the mid-shaft did not show any differences from the normal control subjects. DISCUSSION: This study demonstrates a systemic low BMD including the cortical bone. It is suggested that AIS girls may have disturbance in mineralization and ossification during peripubertal growth.


Subject(s)
Calcification, Physiologic , Scoliosis/physiopathology , Adolescent , Female , Hong Kong , Humans , Tomography, X-Ray Computed
10.
Bone ; 39(4): 863-71, 2006 Oct.
Article in English | MEDLINE | ID: mdl-16765664

ABSTRACT

AIMS: The present study employed both static and dynamic imaging modalities to study both intra- and extravascular events attributing to steroid-associated osteonecrosis (ON) using an experimental protocol with a single low-dose lipopolysaccharide (LPS) injection and subsequently three injections of high-dose methylprednisolone (MPS). METHODS: Fourteen 28-week-old male New Zealand white rabbits received one intravenous injection of LPS (10 microg/kg). 24 h later, three injections of 20 mg/kg of MPS were given intramuscularly at a time interval of 24 h. Additional 6 rabbits were used as controls. Dynamic MRI was performed on bilateral femora for local intraosseous perfusion before and after LPS injection. Blood samples were collected for hematological examinations before and after LPS injection. Bilateral femora were dissected and decalcified for microCT-based microangiography. ON lesion, intravascular thrombus and extravascular marrow fat cell size were examined histopathologically. RESULTS: Intravascular thrombus was observed in all ON rabbits. Extravascular marrow fat cell size was significantly increased in ON rabbits than that of the controls (P<0.05). Compared to baseline, a significant decrease in ratio of tissue-type plasminogen activator/plasminogen activator inhibitor 1, activated partial thromboplatin time and a significant increase in ratio of low-density lipoprotein/high-density lipoprotein were only found in ON rabbits (P<0.05). Dynamic MRI showed a significant decrease in the perfusion index 'maximum enhancement' in the ON rabbits (P<0.05), and microCT-based microangiography showed blocked stem vessels in ON samples. Overall, 93% of the rabbits (13/14) developed ON, and no rabbits died throughout the experiment period. CONCLUSION: Both intra- and extravascular events were found attributing to the steroid-associated ON based on our experimental protocol with a single low-dose LPS injection and subsequent three injections of high-dose MPS. Both high ON incidence and no mortality in rabbits treated with this inductive protocol suggested its effectiveness for future studies on evaluation of therapeutic efficacy of interventions developed for prevention of steroid-associated ON.


Subject(s)
Lipopolysaccharides/toxicity , Methylprednisolone/toxicity , Osteonecrosis/pathology , Animals , Bone Marrow Cells/drug effects , Bone Marrow Cells/pathology , Dose-Response Relationship, Drug , Femur/diagnostic imaging , Femur/drug effects , Femur/pathology , Injections, Intramuscular , Injections, Intravenous , Lipopolysaccharides/administration & dosage , Lipoproteins, HDL/blood , Lipoproteins, LDL/blood , Magnetic Resonance Imaging/methods , Male , Methylprednisolone/administration & dosage , Multivariate Analysis , Osteonecrosis/chemically induced , Partial Thromboplastin Time , Plasminogen Activator Inhibitor 1/blood , Rabbits , Time Factors , Tissue Plasminogen Activator/blood , Tomography, X-Ray Computed/methods
11.
Bone ; 38(6): 818-25, 2006 Jun.
Article in English | MEDLINE | ID: mdl-16413840

ABSTRACT

AIM: Factorial design was used to test our hypothesis whether a group of flavonoids (FE) derived from herbal Epimedium Brevicornum Maxim exerted its preventive effects on estrogen-deficiency-induced osteoporosis mainly through an enhancement in intestinal calcium absorption. MATERIALS AND METHODS: Forty-five 12-month-old female Wistar rats were used and randomly assigned into sham-operated group and four ovariectomy (OVX) subgroups, i.e. OVX with vehicle (OVX group), OVX with FE (FE group), OVX with calcium supplement (CS group), and OVX with FE and CS (FE + CS group). Daily oral administration of FE (10 mg/kg/day) and/or CS (56 mg/kg/day) started on day 4 after OVX for 12 weeks. Before sacrificing the animals, urine and serum samples were collected for assaying indicators related to intestinal calcium absorption, regulator for calcium homeostasis, and markers of bone turnover. The left proximal femur was dissected for evaluation of the primary end-point (failure force), the second end-points (pQCT-quantified densitometry, geometry, and micro-CT-quantified 3-D trabecula micro-architecture), and pQCT-defined cross-sectional envelope. RESULTS: FE was found to be able to prevent OVX-induced reduction in failure force as well as the above second end-points, without resulting in an increased uterus weight. CS had no preventive effect on OVX-induced reduction in failure force. Two-way factorial interaction analysis between FE and CS showed that the un-enhanced suppression of parathyroid hormone for calcium homeostasis did not provide link between the enhanced intestinal calcium absorption and the enhanced inhibition of bone resorption in the present study. Furthermore, the discrepancies between the enhanced intestinal calcium absorption and the un-enhanced end-point measures as well as anabolic effect were also revealed by the interaction analysis. CONCLUSION: The present study suggested that FE inhibited bone resorption, stimulated bone formation, and accordingly prevented osteoporosis without hyperplastic effect on uterus in the OVX rat model, which was however independent of an enhancement in intestinal calcium absorption.


Subject(s)
Calcium/metabolism , Epimedium/chemistry , Flavonoids/pharmacology , Intestinal Absorption/drug effects , Osteoporosis/prevention & control , Ovariectomy , Animals , Body Weight/drug effects , Bone and Bones/cytology , Bone and Bones/drug effects , Bone and Bones/metabolism , Female , Homeostasis , Imaging, Three-Dimensional , Molecular Structure , Organ Size/drug effects , Rats , Rats, Wistar , Stress, Mechanical
12.
Bone ; 37(2): 274-82, 2005 Aug.
Article in English | MEDLINE | ID: mdl-15961358

ABSTRACT

This study used both multi-slice pQCT and microCT to investigate regional changes in bone mineral density and structural parameters in the ultradistal tibia and in the mid-femoral neck under habitual gait loading. Twenty cadavers with 2 females and 18 males aged 70.8 +/- 8.5 were used in this study. Seventy-two cylindrical bone cores with 5 mm in diameter and 10 mm in length from the anterior/posterior and superior/inferior regions were obtained from ultradistal tibia and mid-femur neck, respectively, so that their differences in terms of volumetric trabecular bone mineral density (tBMD) as well as micro-architectural parameters could be studied. The results showed that the mean volumetric tBMD at both the organ (including the bone marrow spaces) and tissue levels (excluding the bone marrow spaces) were a 49.2% and 28.3%, respectively, lower in the anterior bone cores than in the posterior bone cores from the ultradistal tibia (P < 0.01). MicroCT measurements on BV/TV, BS/TV, Tb.N, Tb.Th, and DA were found to be on average of 33.5%, 23.6%, 9.1%, 18.0%, and 14.6%, respectively, lower in the anterior trabecular bone cores (P < 0.001), while Tb.Sp and SMI were 12.5% and 29.3%, respectively, higher in the anterior trabecular bone cores (P < 0.01). No significant difference in micro-architectural parameters was found in the trabecular bone cores obtained from mid-femoral neck, except that the mean DA of the inferior bone cores was significantly higher by 30.1% than that of the superior bone cores (P = 0.01). A statistically significant linear relationship with the correlation coefficient, ranging from 0.37 to 0.94 and -0.62 to -0.85, respectively, was shown between the tBMD at the organ level and all of the micro-architectural parameters (P < 0.05). We suggest that dynamic loading changes during the striking of the heel in normal gait, as well as the peaks of the hip joint reaction force occur during the heel strike and before toe off positions in the lifetime of the subject may account for such regional differences in BMD and micro-architecture. The findings from the correlation study also suggest that, apart from BMD, the micro-architecture may exhibit adaptation in response to such excessive loading.


Subject(s)
Bone Density , Femur/anatomy & histology , Tibia/anatomy & histology , Absorptiometry, Photon , Aged , Cadaver , Female , Femur/diagnostic imaging , Femur/physiology , Humans , Male , Middle Aged , Tibia/diagnostic imaging , Tibia/physiology , Tomography, X-Ray Computed , Weight-Bearing
13.
Biomaterials ; 25(4): 659-67, 2004 Feb.
Article in English | MEDLINE | ID: mdl-14607504

ABSTRACT

The aim of the present study is to demonstrate the newly formed tissue in calcium phosphate (Ca/P) ceramics after extra-osseous implantation by histological and immunohistochemistry (IHC) methods. Synthesis porous Ca/P ceramics without adding any growth factor and living cell were implanted in the dorsal muscle of dogs for 1 and 2 months. Undecalcified and decalcified sections were stained by hematoxylin and eosin (H&E), and IHC, respectively. The histological results showed the beginning of osteogenesis and angiogenesis after being implanted for 1 month and the obvious new bone formation after being implanted for 2 months. IHC were conducted via the avidin-biotin peroxidase complex (ABC) method and the primary antibody was collagen type I. IHC results indicated that collagen type I was expressed within osteoblast-like cells and newly formed bone-like tissue in Ca/P ceramics after 1 month, and in the mineralized matrix of newly formed bone and osteoblasts, some osteocytes and some lacunae after 2 months. No cartilage and chondrocytes were observed in the histological and IHC-stained sections. Evidence of intramembranous osteogenesis was confirmed.


Subject(s)
Bone Substitutes/chemistry , Bone Substitutes/metabolism , Calcium Phosphates/chemistry , Collagen Type I/metabolism , Muscle, Skeletal/cytology , Muscle, Skeletal/physiology , Osteogenesis/physiology , Prostheses and Implants , Animals , Ceramics/chemistry , Dogs , Materials Testing , Muscle, Skeletal/surgery , Neovascularization, Physiologic/physiology , Osseointegration/physiology , Porosity
14.
Chemosphere ; 52(9): 1659-65, 2003 Sep.
Article in English | MEDLINE | ID: mdl-12867200

ABSTRACT

The differential expression of cytochrome P4501A1 gene expression (CYP1A1) in liver, intestine and gill of juvenile tilapia following oral exposure to polychlorinated biphenyl (PCB) Aroclor-1254 or Aroclor-1260 spiked food were investigated. The fish was fed with 0.1 g/g of body weight each day of the spiked food with a concentration of approximately 8 ppm. Fish fed with betaNF- or acetone-spiked food were used as positive and negative controls, respectively. On day 1, 3 and 7 of post-treatment, the fish were sampled and analyzed for the total accumulated levels of PCBs, as well as the induced levels of CYP1A1 mRNA. Our results indicated that the fish fed with PCBs-spiked food showed a time-dependent bioaccumulation of PCBs. The time course studies also revealed that in the first 3 days of feeding, significant increases in CYP1A1 were found in intestine (betaNF: 320% and Aroclor-1260: 290%), followed by liver (betaNF: 144%, Aroclor-1260: 123% and Aroclor-1254: 110%). The transcript levels, then declined slowly. There was no induction of gill CYP1A1 mRNA expression in all treatment groups. Among all the tested tissues, the highest intestinal CYP1A1 transcripts induction has highlighted its prompt response to dietary PCBs exposure. Our study supported the important role of intestine in response to dietary PCBs exposure. In addition, in the present study the relative potency of Aroclor-1260 over Aroclor-1254 in stimulating CYP1A1 expression was discussed, indicating that the highly chlorinated PCBs had a higher potency of bioaccumulation and CYP1A1 mRNA induction.


Subject(s)
Aroclors/pharmacology , Cytochrome P-450 CYP1A1/biosynthesis , Gene Expression Regulation, Enzymologic/drug effects , RNA, Messenger/biosynthesis , Tilapia/metabolism , Animals , Cytochrome P-450 CYP1A1/genetics , Food , Gills/drug effects , Gills/enzymology , Intestines/drug effects , Intestines/enzymology , Liver/drug effects , Liver/enzymology , Polychlorinated Biphenyls/analysis , RNA, Messenger/genetics , Time Factors , Tissue Distribution
15.
J Endocrinol ; 173(1): 199-209, 2002 Apr.
Article in English | MEDLINE | ID: mdl-11927399

ABSTRACT

Stanniocalcin is a polypeptide hormone that was first reported in fish as a regulator of mineral metabolism. Its recent identification in mammals has opened a new area of investigation in basic and clinical endocrinology. In the present study, regulation of the stanniocalcin (STC) and stanniocalcin related protein (STCrP) genes were investigated in mouse neuroblastoma cells (Neuro-2A) in relation to neuronal cell differentiation. Neuro-2A is an undifferentiated cell line that contains measurable levels of STCrP mRNA, but undetectable levels of STC mRNA. Treatment of the cells with either dbcAMP (1-4 mM) or 50 microM euxanthone (PW1) resulted in extensive differentiation and neurite outgrowth. However, only neurites of dbcAMP-treated cells developed varicosities, a phenotypic marker of axon formation. Furthermore, following differentiation induced by dbcAMP, there was an upregulation of STC and downregulation of STCrP mRNA levels. In the first 24 and 48 h of treatments, there was a maximum twofold induction and 1.5-fold reduction in STC and STCrP mRNAs respectively. Following 96 h of treatment, an additional 14-fold STC induction and 1.2-fold STCrP reduction were observed. The increase in STC mRNA levels was accompanied by a concomitant increase in axon-specific low molecular form microtubule-associated protein (MAP-2c) mRNA and varicosities on the neurites, suggesting a possible role for STC in axonogenesis. There was no induction of STC mRNA levels when PW1 was added into the culture media, whereas ionomycin (1-10 microM) had no observable effects on cell differentiation or STC/STCrP mRNA. Immunocytochemical staining of dbcAMP-treated cells revealed abundant levels of immunoreactive STC, particularly in the varicosities, with only weak staining in control, untreated cells. Antisense oligodeoxynucleotides transfection studies indicated that the expression of STC was a cause of varicosity formation and a consequence of cell differentiation. Our findings lend further support to the notion that STC is involved in the process of neural differentiation.


Subject(s)
Bucladesine/pharmacology , Gene Expression Regulation/drug effects , Glycoproteins/genetics , Hormones/genetics , Neurons/drug effects , RNA, Messenger/metabolism , Xanthones , Animals , Bucladesine/analysis , Cell Differentiation/drug effects , Dose-Response Relationship, Drug , Ionomycin/pharmacology , Ionophores/pharmacology , Mice , Neurites/drug effects , Neuroblastoma , Neurons/cytology , Oligonucleotides, Antisense/pharmacology , RNA, Messenger/analysis , Stimulation, Chemical , Time Factors , Tumor Cells, Cultured , Xanthenes/pharmacology
16.
Aquat Toxicol ; 54(1-2): 69-80, 2001 Sep.
Article in English | MEDLINE | ID: mdl-11451427

ABSTRACT

Toxicological effects of persistent organic pollutants (POPs) in aquatic ecosystems lead to the deterioration of water quality and adversely affect fish and human health. The highly lipophilic nature of these pollutants may enter fish through the diet or by water-borne exposure. In monitoring contamination in aquatic systems, induction of the cytochrome P450 1A1 (CYP1A1) gene of fish has been evaluated as a sensitive, "early warning" method. The objective of the present study was to characterize the induction of the gene in fish upon exposure to coastal sediments and to determine its specific expression in liver and extrahepatic organs (i.e. gill and intestine) in which the toxicological effects to the corresponding tissues could be addressed. Sediment samples were collected from different sites, including Victoria Harbour (VS), Ma Wan (MW), Tsim Bei Tsui (TBT) and Mai Po (MP). The samples were analyzed for polychlorinated biphenyls (PCBs) and polycyclic aromatic hydrocarbons (PAHs). CYP1A1 mRNA expression was measured in juvenile tilapia exposed experimentally to coastal sediment for 3 and 7 days. A negative control group of fish maintained in seawater was used. Using the primer dropping polymerase chain reaction technique, gill, intestinal and hepatic CYP1A1 mRNAs were quantified. Chemical analysis shows that the samples from VS contaminated with the highest concentration of PCBs (45.24 p.p.b.) and PAHs (1663.7 p.p.b.), followed by MW (16.01 and 347.7 p.p.b.), TBT (14.48 and 235.2 p.p.b.) and MP (14.60 and 242.2 p.p.b.). Fish exposed to sediments were contaminated with various levels of PCBs (VS, 64.14-72.06 p.p.b.; MP, 27.06-31.62 p.p.b.; TBT, 27.29-33.92 p.p.b.; MW, 16.05-17.76 p.p.b.) and PAHs (VS, 124.7-304.9 p.p.b.; MP, 97.57-164.1 p.p.b.; TBT, 25.38-98 p.p.b.; MW, 24.07-68.13 p.p.b.). The control fish displayed moderate expression of CYP1A1 mRNA in liver (1.45 arbitrary units), gill (1.21 arbitrary units) and intestine (0.56 arbitrary units). Following sediment exposure, there was a large increase in CYP1A1 mRNA in intestine and liver but no significant changes in gill tissues. In the first 3 days, there was anywhere from 34-96 and 69-156% induction of the CYP1A1 transcripts in intestine and liver, respectively. Following 7 days of exposure, a continued induction of high level of CYP1A1 mRNA in intestine (73-157%) was observed. The induction of CYP1A1 in liver and intestine provided a defensive mechanism against POPs entering from the external environment.


Subject(s)
Cytochrome P-450 CYP1A1/genetics , Gene Expression Regulation, Enzymologic/drug effects , Geologic Sediments , Gills/enzymology , Intestines/enzymology , Liver/enzymology , Tilapia/metabolism , Water Pollutants, Chemical/toxicity , Animals , Polychlorinated Biphenyls/analysis , Polycyclic Aromatic Hydrocarbons/analysis , RNA, Messenger/analysis
17.
Clin Orthop Relat Res ; (385): 219-29, 2001 Apr.
Article in English | MEDLINE | ID: mdl-11302317

ABSTRACT

Distraction osteogenesis is a process of tissue regeneration under an external mechanical stimulation. In the study, the spatial and temporal expression patterns of basic fibroblast growth factor in the newly formed osseous tissue during distraction osteogenesis in goats were studied using immunohistochemistry. During the distraction period, the expression of basic fibroblast growth factor was observed in the osteoblasts on the newly formed trabecular bone and the bone formation front. The cells of osteoblastic lineage and the mesenchymal cells in the distraction callus also expressed basic fibroblast growth factor. The expression of basic fibroblast growth factor in the distraction period was stronger than that during the latency and consolidation periods. However, some osteoblasts still were expressing basic fibroblast growth factor in the consolidation periods. According to these results, basic fibroblast growth factor may have a local regulatory role during distraction osteogenesis. The tensile force may stimulate the expression of basic fibroblast growth factor in osteoblasts and other cell types.


Subject(s)
Bone and Bones/metabolism , Fibroblast Growth Factor 2/metabolism , Osteogenesis, Distraction , Animals , Bony Callus/pathology , Goats , Immunohistochemistry , Models, Animal , Osteoblasts/pathology
18.
Arch Environ Contam Toxicol ; 38(4): 486-93, 2000 May.
Article in English | MEDLINE | ID: mdl-10787100

ABSTRACT

The aim of the present project is to determine the feasibility of measuring hepatic cytochrome P4501A1 (CYP1A1) and metallothionein (MT) mRNA in fish as an integrative measurement of persistent organic pollutants (POPs) and heavy metal contamination in sediment arising in Hong Kong. Sediment samples were collected from different sites, including Victoria Harbour (VS6), Yim Tin Tsai (YTT) at Tolo Harbour, Mai Po marshes (MPM) at Deep Bay, and Southern Waters (SS6) of coastal waters. The samples were analyzed for total and extractable concentrations of Cd, Cu, Ni, Zn, and Pb, as well as PCBs and PAHs. In addition, biomarker responses were studied in tilapia exposed experimentally to coastal sediment for 7 days. Using RT-PCR technique, hepatic CYP1A1 and MT mRNA were measured. Three control groups were used, including one negative control group maintained in sea water only; the second and third positive control groups were in sea water but were intraperitoneally injected with either beta-naphthoflavone (40 microg/g body weight) or cadmium chloride (10 microg/g body weight), respectively. The chemical data showed that VS6, YTT, and MPM were classified as Class C sediment according to the sediment quality criteria defined by the Hong Kong Environmental Protection Department, indicating the sites were heavily polluted. The exposure of tilapia to the sediment induced hepatic CYP1A1 (VS6 > YTT > MPM > SS6) and MT (VS6 > MPM > YTT > SS6) levels. The induction patterns were comparable to the levels of POPs and metal contamination in the sediment, indicating that the biomarker responses could be used to differentiate low to high levels of contamination among sediment.


Subject(s)
Geologic Sediments/chemistry , Metals, Heavy/analysis , Polychlorinated Biphenyls/analysis , Polycyclic Aromatic Hydrocarbons/analysis , Water Pollutants, Chemical/analysis , Animals , Cadmium Chloride/toxicity , Cytochrome P-450 CYP1A1/genetics , Cytochrome P-450 CYP1A1/metabolism , DNA/analysis , DNA Primers/chemistry , Evaluation Studies as Topic , Hong Kong , Liver/enzymology , Metallothionein/genetics , Metallothionein/metabolism , RNA, Messenger/biosynthesis , Reverse Transcriptase Polymerase Chain Reaction , Tilapia/metabolism , beta-Naphthoflavone/toxicity
19.
J Vasc Surg ; 6(4): 419-21, 1987 Oct.
Article in English | MEDLINE | ID: mdl-3656589

ABSTRACT

We report a case of an 11-year-old girl with a pulsatile mass in the thigh diagnosed by angiography as localized arteriovenous malformation. Recent increase in tenderness and size of the lesion brought the patient to hospital and excision was performed at the parents' request. Subsequent histologic examination revealed that this lesion was an alveolar soft-part sarcoma in disguise. The angiographic and pathologic results are presented. Other investigators should be aware of this uncommon and highly vascular sarcoma, which might be misdiagnosed as a localized arteriovenous malformation and thus treated only conservatively.


Subject(s)
Arteriovenous Malformations/diagnosis , Sarcoma/diagnosis , Soft Tissue Neoplasms/diagnosis , Thigh , Arteriovenous Malformations/diagnostic imaging , Arteriovenous Malformations/surgery , Child , Diagnostic Errors , Female , Humans , Radiography , Sarcoma/pathology , Soft Tissue Neoplasms/pathology , Thigh/blood supply
20.
Cancer ; 57(12): 2381-8, 1986 Jun 15.
Article in English | MEDLINE | ID: mdl-3084061

ABSTRACT

A case of angiosarcoma (malignant hemangioendothelioma) developing in a chronic goitrous thyroid gland of an elderly Chinese woman is described. Histologically it showed the same classical appearance of angiosarcoma occurring in the skin and soft tissue. The endothelial origin of this tumor was confirmed by demonstrating Factor VIII-related antigen in the neoplastic cells with the immunoperoxidase technique and Weibel-Palade bodies by electron microscopic study. Because of its extreme rarity outside the European Alpine regions, many authorities are reluctant to accept it as a distinct entity and merely consider it as a variant of an undifferentiated carcinoma. Our report not only provides additional evidence that angiosarcoma of the thyroid gland is a specific condition of endothelial origin but also documents the first case among Chinese.


Subject(s)
Hemangiosarcoma/ultrastructure , Thyroid Neoplasms/ultrastructure , Aged , Antigens/analysis , Asian People , China , Factor VIII/analysis , Factor VIII/immunology , Female , Hemangiosarcoma/metabolism , Humans , Immunoenzyme Techniques , Microscopy, Electron , Thyroid Neoplasms/metabolism , von Willebrand Factor
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