摘要
Objective To prepare pH sensitive gel for eye cornea of salvianolic acid B and accomplish in vitro release study. Me thods We used corneal permeability, eye irritation in rabbits, hydration of the cornea, viscosity, gel capacity, transparency, liquidity as index, to select pH sensitive gel matrix composition. Re s ults The gel composition was: 0.5% of salvianolic acid B, 0.14% Carbomer, 0.3% HPMC, 0.05% Azone, 0.1% ethylparaben and 0.2% Vc. The corneal hydration level was 77.32±1.88.The gel released 27.21% drug in 15 min, and released 97.44% drug in 6 h. By research of corneal permeability, the cumulative release of gel was bigger than the one of saline (P<0.05). Conclus ions The selection of the gel matrix prescription is reasonable in design, simple in process, not harsh to eyes, and can prolong the retention time of drug in corneas and improve the corneal penetration.
摘要
Recently, activation of the adenosine A2A receptors has been shown to exert protection against peripheral tissue injuries but aggravation in the central nervous system (CNS) injuries. To explore the different effects of adenosine A2A receptors and try to perform some new treatment strategies for peripheral tissue and CNS traumas, we constructed the mouse models of skin trauma, skin combined radiation-impaired wound and traumatic brain injury (TBI), respectively. Wild type mice and A2A receptor gene knockout mice were both used in the experiments. In skin trauma and combined radiation-impaired wound models, the time of wound healing was observed, while in TBI model, neurological deficit scores, water content in injured brain and glutamate concentration in cerebral spinal fluid (CSF) were detected at 24 h after TBI. The results showed that in skin trauma and combined radiation-impaired wound models, CGS21680 (an agonist of the A2A receptors) promoted while A2A receptor gene knockout delayed the course of skin wound healing. On the contrary, in TBI model, A2A receptor gene knockout, not CGS21680, showed a protective role by inhibition of glutamate release. These data further indicate that promoting glutamate release may account for the different effects of A2A receptor activation in CNS injury and peripheral tissue injury models. These findings may provide some experimental evidence and a new strategy for clinical treatment of peripheral tissue damages by agonists of A2A receptors, while treatment of CNS injuries by antagonists of A2A receptors.
Subject(s)
Animals , Mice , Adenosine , Pharmacology , Brain , Pathology , Brain Injuries , Disease Models, Animal , Glutamic Acid , Cerebrospinal Fluid , Mice, Knockout , Phenethylamines , Pharmacology , Receptor, Adenosine A2A , Genetics , Physiology , Wound Healing摘要
Burn-blast combined injury is caused by two injury factors--heat and blast, which inflict the body at the same time or in sequence. The incidence of the combined injury is high either in wartime or in peacetime, and the mortality is much higher than that of an injury due to either one injury factor. In order to elucidate the mechanism, characteristics of the injury and the treatment of the combined injury, lots of studies were carried out both at home and abroad. The paper presents the data of burn-blast injury from a part of experimental studies and some clinical experience in the past forty years. The paper may be useful to medical doctors who may treat burn-blast injury in future.
Subject(s)
Animals , Humans , Blast Injuries , Therapeutics , Burns , Therapeutics , Multiple Trauma , Therapeutics摘要
<p><b>OBJECTIVE</b>To study the expression regularity of vascular endothelial growth factor (VEGF) during the process of fracture healing, and the type of VEGF receptor expressed in the vascular endothelial cells of the fracture site.</p><p><b>METHODS</b>The fracture model was made in the middle part of left radius in 35 rabbits. The specimens from the fracture site were harvested at 8, 24, 72 hours and 1, 3, 5, 8 weeks, and then fixed, decalcified, and sectioned frozenly to detect the expression of VEGF and its receptor at the fracture site by in situ hybridization and immunochemical assays.</p><p><b>RESULTS</b>VEGF mRNA and VEGF expression was detected in many kinds of cells at the fracture site during 8 hours to 8 weeks after fracture. Flt1 receptor of VEGF was found in the vascular endothelial cells at the fracture site during 8 hours to 8 weeks after fracture, and strong expression of flk1 receptor was detected from 3 days to 3 weeks after fracture.</p><p><b>CONCLUSIONS</b>The expression of VEGF and flt1 receptor appears during the whole course of fracture healing, especially from 1 to 3 weeks. Flk1 receptor is highly expressed in a definite period after fracture. VEGF is proved to be involved in the vascular reconstruction and fracture healing.</p>
Subject(s)
Animals , Female , Male , Rabbits , Endothelial Cells , Chemistry , Fracture Healing , Physiology , Immunohistochemistry , In Situ Hybridization , Receptors, Vascular Endothelial Growth Factor , Vascular Endothelial Growth Factor A摘要
<p><b>OBJECTIVE</b>To establish a mice model with selective inactivation adenosine A2A receptors (A2ARs) in peripheral white blood cells (PWBC).</p><p><b>METHODS</b>A2ARs were selectively inactivated in PWBCs by transplanting bone marrow cells (BMCs) from A2AR knockout (KO) mice into their wild type (WT) littermates after a single total body irradiation of 9.5 Gy or fractionated total body irradiation of 6.2 Gy x 2. The efficiency of reconstitution of bone marrow-derived cells in chimeric mice was assessed.</p><p><b>RESULTS</b>PCR band patterns changed from the recipient pattern (one band of 330 bp) to the donor (two bands of 300 and 330 bp) pattern. Immunohistochemistry analysis showed that 10.21% of cells were A2AR+ in PWBCs in KO--> WT mice, whereas 96.72% of cells were A2AR+ in WT mice. The survival rates of mice irradiated with 6.2 Gy x 2 and transplanted with more than 6 x 10(6) BMCs were about 91%.</p><p><b>CONCLUSION</b>A murine model of selective inactivation adenosine A2A receptors in PWBCs was established successfully.</p>
Subject(s)
Animals , Mice , Gene Deletion , Mice, Inbred C57BL , Mice, Knockout , Models, Animal , Receptor, Adenosine A2A , Genetics摘要
<p><b>OBJECTIVE</b>To investigate the polymorphisms of myeloid differentiation-2 (MD-2) gene promoters, and to explore whether such polymorphisms are associated with the susceptibility to multiple organ dysfunction syndrome (MODS) and sepsis in Chinese Han population.</p><p><b>METHODS</b>Using polymerase chain reaction-restriction fragment length polymorphism method, the authors detected the single nucleotide polymorphisms of the promoter region of MD-2 gene at position - 1625C/G in 105 severe trauma patients (42 with sepsis). The organ function was scored.</p><p><b>RESULTS</b>The frequency of CC genotype in MD-2 gene promoter region at position - 1625 was 0.5 (21/42) in septic patients and 0.7 (44/63) in non-septic patients. The frequency of CG genotype was 0.38 (16/42) in septic patients and 0.27 (17/63) in non-septic patients. The frequency of GG genotype was 0.12 (5/42) in septic patients and 0.03 (2/63) in non-septic patients. The MODS scores in trauma patients carrying G allele at position - 1625 were significantly higher than those carrying C allele (P<0.001 for dominant effect, and P>0.05 for recessive effect). Moreover, trauma patients carrying G allele appeared to have higher risk of sepsis comparing to those carrying C allele (OR 0.477, 95% CI 0.266-0.855, P<0.05). Sepsis morbidity was significantly different between subjects with C and G alleles (P<0.05 for dominant effect, P>0.05 for recessive effect).</p><p><b>CONCLUSIONS</b>The polymorphisms of the promoter region of MD-2 gene at position - 1625 C/G is correlated with MODS and sepsis after severe trauma in Chinese Han population. The people with - 1625 G allele in the promoter region of MD-2 gene may be a risk factor of severe complications.</p>
Subject(s)
Humans , Asian People , China , Genetic Predisposition to Disease , Lymphocyte Antigen 96 , Genetics , Multiple Organ Failure , Genetics , Polymorphism, Genetic , Promoter Regions, Genetic , Sepsis , Genetics , Wounds and Injuries , Genetics摘要
<p><b>OBJECTIVE</b>To study the effect of vascular endothelial growth factor (VEGF)and anti-VEGF on the expression of fracture healing-related factors and observe pathological changes at fractured sites.</p><p><b>METHODS</b>Fracture models were established in 105 New Zealand white rabbits and they were randomly divided into control group, VEGF group and anti-VEGF group. The relevant factors expression at fractured sites was assayed and pathological changes were observed in decalcified samples at 8, 24, 72 hours and 1,3,5,8 weeks after fracture.</p><p><b>RESULTS</b>After application of VEGF, the expression of BMP appeared earlier and expression time lasted longer. On the contrary, anti-VEGF completely inhibited the expression of BMP. The fractured sites were filled with fibrous callus, cartilaginous callus and bony callus at the 3rd week and woven bone was constructed at the 5th week. Fracture healing was accomplished at the 8th week in VEGF group. In anti-VEGF polyclonal antibody group, cellular necrosis increased at early period. Continuous focal necrosis was seen in the fractured sites from the 1st week to 5th week. Vascularization reduced obviously at the 3rd week.</p><p><b>CONCLUSIONS</b>Fracture healing is a result of mutual regulation and coordination among many factors. VEGF may be an important factor in fracture healing.</p>
Subject(s)
Animals , Rabbits , Bone Morphogenetic Proteins , Metabolism , Electrophoresis, Polyacrylamide Gel , Fibroblast Growth Factor 2 , Metabolism , Fracture Healing , Physiology , Radius Fractures , Vascular Endothelial Growth Factor A , Physiology摘要
<p><b>OBJECTIVE</b>In order to investigate the evidence of the synergistic effects of bacterial components, to observe the relationship of the expression of lipopolysaccharide (LPS) receptors [CD14, Toll-like receptor 4 (TLR4), scavenger receptor (SR)], lipoprotein receptor (TLR2) and bacterial DNA receptor (TLR9) with pulmonary injury in abdominal infection-induced sepsis.</p><p><b>METHODS</b>30 mice were used and randomly divided into cecal ligation puncture (CLP) (n = 15) and sham (n = 15) groups. The animals were respectively sacrificed 8, 12 and 24 (each point n = 5) hours following CLP and sham CLP. The lungs were removed and immediately stored in liquid nitrogen for TLRs mRNA, tumor necrosis factor (TNF) alpha and myeloperoxidase (MPO) assay. To detect the expression of CD14, TLR4, SR, TLR2 and TLR9 mRNA by reverse-transcription polymerase chain reaction, to detect the TNF-alpha content of the lung tissue by enzyme-labeled immunosorbent assay, and to assay the MPO activity of the lung tissue spectrophotometer.</p><p><b>RESULTS</b>It was found that the expression of receptors for LPS, BLP and bacterial DNA in pulmonary tissues was markedly changed in CLP-induced sepsis, showing upregulation of CD14 mRNA (1.143 +/- 0.139, t = 0.022, P < 0.05), TLR2 mRNA (0.418 +/- 0.102, t = 0.021, P < 0.05), TLR4 mRNA (0.595 +/- 0.052, t = 0.0001, P < 0.01) and TLR9 mRNA (0.743 +/- 0.178, t = 0.0023, P < 0.01) at different degrees (P < 0.05 or P < 0.01) after postinjury 8 h, among which the expression of TLR9 mRNA kept increasing. The expression of SR mRNA (8 h: 0.659 +/- 0.159; 12 h: 0.429 +/- 0.061; 24 h: 0.300 +/- 0.045; t = 0.029, P < 0.05; t = 0.001, P < 0.01; t = 0.003, P < 0.01) showed continuous down-regulation.</p><p><b>CONCLUSION</b>There was a marked correlation between the changes of pattern-recognition receptor expression and the increases of MPO and TNF-alpha levels in pulmonary tissues.</p>
Subject(s)
Animals , Mice , Disease Models, Animal , Lung , Metabolism , Pathology , Mice, Inbred Strains , RNA, Messenger , Genetics , Receptors, Pattern Recognition , Genetics , Sepsis , Metabolism , Pathology , Tumor Necrosis Factor-alpha , Metabolism摘要
<p><b>OBJECTIVE</b>To explore the protective effects of earplug and barrel on auditory organs of guinea pigs exposed to experimental blast underpressure (BUP).</p><p><b>METHODS</b>The hearing thresholds of the guinea pigs were assessed with auditory brainstem responses (ABR). The traumatic levels of tympanic membrane and ossicular chain were observed under stereo-microscope. The rate of outer hair cells (OHCs) loss was analyzed using a light microscope. The changes of guinea pigs protected with barrel and earplug were compared with those of the control group without any protection.</p><p><b>RESULTS</b>An important ABR threshold shift of the guinea pigs without any protection was detected from 8h to 14d after being exposed to BUP with a peak ranging from -64.5 kPa to -69.3 kPa ( P<0.01). The rate of perforation of tympanic membrane reached 87.5% and that of total OHCs loss was 19.46% +/- 5.38% at 14d after exposure. The guinea pigs protected with barrel and earplug had lower ABR threshold and total OHCs loss rate compared with the animals without any protection (P<0.01). All of the tympanic membrane and ossicular chain of the protected animals maintained their integrities. Meanwhile, the guinea pigs protected with the barrel had lower ABR threshold and total OHCs loss rate than those with earplug (P<0.01).</p><p><b>CONCLUSIONS</b>The earplug and barrel have protective effects against BUP-induced trauma on auditory organs of the guinea pigs and the protective effects of barrel are better than those of earplug.</p>
Subject(s)
Animals , Auditory Threshold , Blast Injuries , Ear Protective Devices , Guinea Pigs , Hair Cells, Auditory, Outer , Metabolism , Pressure , Tympanic Membrane , Wounds and Injuries , Tympanic Membrane Perforation摘要
<p><b>OBJECTIVE</b>To investigate the expression of myeloid differentiation protein-2 (MD-2) in the human endothelial cells and its role in lipopolysaccharide (LPS)-induced NF-kappaB activation in endothelial cells.</p><p><b>METHODS</b>In vitro cultured human umbilical vein endothelial cells (HUVEC) were employed in the study. The expression of MD-2 mRNA and protein, and the effect of LPS on the expression of its mRNA and protein were assessed with RT-PCR and Western blotting. The role of MD-2 in LPS-induced NF-kappaB activation and IL-8 production were investigated with gene transfection of mutant MD-2 cDNA (0.5, 1.0, 2.0 microg), pEF-BOS vacant vector (2.0 microg) and MD-2 plasmid (2.0 microg) into HUVEC, respectively.</p><p><b>RESULTS</b>There was MD-2 mRNA and protein expression in HUVECs before LPS stimulation, and it could be obviously upregulated by LPS in time and dose-dependent manner (MD-2 protein absorbency was 25 196 +/- 1 723 without LPS stimulation, which was obviously lower than that stimulated with 0.01 mg/L LPS (58 817 +/- 3 241, P < 0.01) for 6 hours. Transfection of mutant MD-2 cDNA could remarkably inhibit LPS-induced NF-kappaB activation and IL-8 production in endothelial cells.</p><p><b>CONCLUSION</b>MD-2 might play an important role in the LPS-induced NF-kappaB activation in HUVECs.</p>
Subject(s)
Humans , Cell Differentiation , Cells, Cultured , Endothelial Cells , Metabolism , Interleukin-6 , Genetics , Interleukin-8 , Metabolism , Lipopolysaccharides , Pharmacology , NF-kappa B , Metabolism , Umbilical Veins , Cell Biology摘要
<p><b>OBJECTIVE</b>To investigate the effects of tumor necrosis factor alpha (TNF alpha) and interferon gamma (IFN gamma) on the expression of pattern recognition receptors (PRRs) on the surface of mouse alveolar macrophages.</p><p><b>METHODS</b>Alveolar macrophages from mouse were cultured in DMEM supplemented with 10% (V/V) endotoxin-free calf serum. After the alveolar macrophages were stimulated with TNF alpha and IFN gamma (concentration, 20 ng/ml) for 3 h, 6 h and 12 h, the expression of PRRs, including cluster of differentiation 14 (CD14), scavenger receptor (SR), toll-like receptor 4 (TLR4), TLR2 and TLR9 mRNA and proteins were examined by RT-PCR and immunohistochemistry.</p><p><b>RESULTS</b>The expressions of CD14, TLR2 and TLR9 receptors, which were related with cellular activation, were up-regulated by the stimulation of TNF alpha and IFN gamma (P < 0.05), while SR, which was related with cellular defense action, was down-regulated (P < 0.05). Although the expression of TLR4 was up-regulated, there was no statistical significance (P > 0.05).</p><p><b>CONCLUSIONS</b>The cytokines such as TNF alpha and IFN gamma could also produce feedback regulation on the expression of PRRs at the levels of genes and proteins. Such regulation on the PRRs expression would be significant for further amplification of inflammation cascade and eventually leading to uncontrolled inflammation.</p>
Subject(s)
Animals , Mice , Cells, Cultured , Interferon-gamma , Pharmacology , Lipopolysaccharide Receptors , Genetics , Macrophages, Alveolar , Metabolism , RNA, Messenger , Genetics , Receptors, Pattern Recognition , Toll-Like Receptor 2 , Genetics , Toll-Like Receptor 4 , Genetics , Toll-Like Receptor 9 , Genetics , Tumor Necrosis Factor-alpha , Pharmacology摘要
<p><b>OBJECTIVE</b>To identify the single nucleotide polymorphisms(SNPs) in the regulatory and coding regions of human Toll-like receptor 4(TLR4) gene and to search for its new genetic makers.</p><p><b>METHODS</b>The 5' flank region, exons, parts of the introns, as well as 3' flank region of TLR4 gene were sequenced to identify and characterize the SNPs in Chinese population. SNP genotyping was performed by polymerase chain reaction-restriction fragment length polymorphism for 2 highly distributed SNPs.</p><p><b>RESULTS</b>Five novel SNPs were identified through a 4.98 kb sequencing of TLR4 gene. Among them, three were in 5'flank region, two in 3'UTR. In the sample of Han population from Chongqing, the minor allele frequencies of two highly distributed SNPs were 0.266 and 0.404 respectively.</p><p><b>CONCLUSION</b>Sampling analysis in Han population of Chongqing showed that the two highly distributed SNPs of TLR4 were common in Chinese population and could be used for genetic marker of TLR4 gene.</p>
Subject(s)
Adult , Female , Humans , Male , Middle Aged , Asian People , Genetics , Base Sequence , China , Gene Frequency , Polymerase Chain Reaction , Polymorphism, Restriction Fragment Length , Polymorphism, Single Nucleotide , Sequence Analysis, DNA , Toll-Like Receptor 4 , Genetics摘要
<p><b>OBJECTIVE</b>To investigate the therapeutic effect of cisapride on gastric injury following hemorrhagic shock resuscitation.</p><p><b>METHODS</b>108 Wistar rats weighing (200 g+/-30 g) were randomly divided into a sham shock (SS) group (n=36), a hemorrhagic shock resuscitation (HS) group (n=36) and a hemorrhagic shock cisapride treated (HSC) group (n=36). Sampling at 1, 2 and 4 hours after resuscitation was done and 6 samples for each observation item were taken. The gastric blood flow volume was measured by isotope label biological microglobulin. Gastric pHi, gastric emptying, MDA and Na+-K+-ATPase of gastric mucosa were measured.</p><p><b>RESULTS</b>In the HSC group, the relative residual rate of gastric pigment decreased significantly, the gastric blood flow volume elevated; gastric pHi increased significantly at 2 hours; the level of mucosal MDA decreased at 4 hours, the activity of Na+-K+-ATPase increased and the lactic acid level in the portal vein decreased significantly compared to the HS group.</p><p><b>CONCLUSIONS</b>After hemorrhagic shock resuscitation, cisapride contained the following functions, 1) promoting gastric emptying, 2) increasing the blood flow of gastric blood flow volume and gastric pHi, 3) depressing the lactic acid concentration of the portal vein and improving MDA volume and Na+-K+-ATPase activity of gastric mucosa. It suggests that after complementing effective circulating blood volume for hemorrhagic shock resuscitation, early use of cisapride for gastric motility is helpful for an improvement of lasting ischemia and hypoxia in stomach.</p>
Subject(s)
Animals , Rats , Cisapride , Therapeutic Uses , Disease Models, Animal , Gastric Emptying , Gastric Mucosa , Chemistry , Gastrointestinal Agents , Therapeutic Uses , Hydrogen-Ion Concentration , Rats, Wistar , Regional Blood Flow , Resuscitation , Shock, Hemorrhagic , Therapeutics , Sodium-Potassium-Exchanging ATPase , Metabolism , Stomach , Pathology , Stomach Diseases , Drug Therapy摘要
<p><b>OBJECTIVE</b>To explore the expression of thrombospondin 2 (TSP2) during the repair process after alkali burn injury to cornea in mice.</p><p><b>METHODS</b>Forty mice were employed in the study. The murine corneal alkali burn model was reproduced (n = 35) as experimental group (E), and the mice were randomly divided into control (C, n = 5) and experimental (E, n = 35) groups. The mice in E group were again divided into 7 sub-groups according to different time points [3, 6, 12, 24, 48, 96 and 192 postburn hours (PBHs)] with 5 mice in each sub-group. HE staining, immunocytochemistry (ICC) and RT-PCR were employed to observe the expression of TSP2 in the corneal tissue of mice in both control and all animals in experimental sub-groups at all above mentioned time points (PBHs).</p><p><b>RESULTS</b>TSP2 was expressed in corneal tissue in both C and E groups, especially in the basal layer of epithelial layer, and also a weak expression in substantia propria layer. Compared with that in C group (0.48 +/- 0.15), the expression of TSP2 in E group enhanced at 3 PBH, peaked at 6 PBH, (1.54 +/- 0.45, P < 0.05), dropped to the nadir at 24 PBH (0.73 +/- 0.19), and bounded back afterwards. It peaked again at 96 PBH (1.79 +/- 0.63, P < 0.05), then decreased thereafter, and approached the control level at 192 PBH (P > 0.05). There was remarkable angiogenesis in the cornea at 24 PBH in the mice in E group.</p><p><b>CONCLUSION</b>The expressions of TSP2 exhibits fluctuating changes along with the course of repair. This might be related to the compensatory process under stress condition.</p>
Subject(s)
Animals , Mice , Burns, Chemical , Metabolism , Pathology , Cornea , Metabolism , Corneal Injuries , Eye Burns , Metabolism , Pathology , Gene Expression , Mice, Inbred Strains , Thrombospondins , Wound Healing摘要
<p><b>OBJECTIVE</b>To explore the proliferation-promoting effect of sensory neuropeptide substance P (SP) on the cultured granulation tissue fibroblasts in vitro and its regulative effect on the gene expression of basic fibroblast growth factor (bFGF) mRNA.</p><p><b>METHODS</b>The proliferation-promoting effect of cultured granulation tissue fibroblasts was observed by means of MTT; the regulative effect of SP on gene expression of fibroblast bFGF by RT-PCR. The time and dose-efficiency relations were also observed.</p><p><b>RESULTS</b>There was a significant proliferation-promoting effect of SP on the cultured granulation tissue fibroblasts in vitro in a remarkable dose-dependent fashion. However, bFGF antibody only partly exerted its inhibitive effect. SP could induce the bFGF mRNA expression of the fibroblasts at the 3rd and 6th hour (P < 0.01). SP could promote the bFGF mRNA expression of the fibroblasts in the concentration of 10(-9) - 10(-5) mol/L and peaked in the concentration of 10(-7) mol/L.</p><p><b>CONCLUSIONS</b>SP has a significant proliferation-promoting effect on the granulation tissue fibroblasts, which is correlated with SP inducing bFGF mRNA expression of fibroblasts.</p>
Subject(s)
Animals , Male , Rats , Cell Division , Dose-Response Relationship, Drug , Fibroblast Growth Factor 2 , Genetics , Fibroblasts , Cell Biology , Metabolism , Gene Expression , Granulation Tissue , Cell Biology , Metabolism , RNA, Messenger , Genetics , Metabolism , Rats, Wistar , Reverse Transcriptase Polymerase Chain Reaction , Substance P , Pharmacology摘要
<p><b>OBJECTIVE</b>To investigate the frequencies of -31, -511 and -1470 single nucleotide polymorphisms (SNPs) in the promoter of IL-1 beta in Chongqing population and address the question whether they are in linkage disequilibrium (LD).</p><p><b>METHODS</b>One hundred and twelve healthy Chongqing people were enrolled in this study. Polymorphisms at -31 (C>T), -511 (T>C) and -1470 (G>C) of IL-1 beta were genotyped with the method of restriction fragment length polymorphism (RFLP). Haplotype frequencies were evaluated with Arlequine software. Two-point LD analyses were done with the software TransposerV1-0. The P values were determined by Pearson chi square test analysis.</p><p><b>RESULTS</b>Allelic frequencies of IL-1 beta -31, -511 and -1470 were 45.33%, 50.00% and 41.67%, respectively. The dominant haplotypes comprising the three loci were T-C-G (44.1%), C-T-C(40.3%) and C-T-G(8.8%), LD analyses revealed that none of the LD parameters(delta value) was 0. Meanwhile, chi square test showed P<0.005.</p><p><b>CONCLUSION</b>-31, -511 and -1470 loci in the promoter region of IL-1 beta are in strong linkage disequilibrium. And this study provides a basis for searching disease-related IL-1 beta haplotye.</p>
Subject(s)
Adult , Female , Humans , Male , Middle Aged , China , Gene Frequency , Genotype , Haplotypes , Interleukin-1beta , Genetics , Linkage Disequilibrium , Polymorphism, Restriction Fragment Length , Polymorphism, Single Nucleotide , Promoter Regions, Genetic , Genetics摘要
<p><b>OBJECTIVE</b>To identify the single nucleotide polymorphisms (SNPs) in the regulatory and coding regions of human interleukin-1 receptor type I (IL-1R1) gene and to assess their potential effect on the function of IL-1R1.</p><p><b>METHODS</b>The 5' flank region, exons, parts of the introns, as well as 3' flank region of IL-1R1 gene were sequenced to identify and characterize the SNPs in Chinese population. Effects of the SNP on the structure and function of IL-1R1 were analyzed by computational methods.</p><p><b>RESULTS</b>Sixteen SNPs were identified through a 9643 bp sequencing of IL-1R1 gene. Among them, four were in 5' flank region, four in intron region, one in coding region, and seven in 3' untranslated region. A novel SNP in Chinese population was involved in a structural change in IL-1R1, which may influence the signal transduction of IL-1R1.</p><p><b>CONCLUSION</b>The SNP in the IL-1R1 gene might influence its function as an important receptor of IL-1 family.</p>
Subject(s)
Humans , Amino Acid Sequence , Asian People , Cell Membrane , Metabolism , Computational Biology , Exons , Genetics , Hydrophobic and Hydrophilic Interactions , Introns , Genetics , Molecular Sequence Data , Polymorphism, Single Nucleotide , Protein Structure, Secondary , Protein Structure, Tertiary , Receptors, Interleukin-1 Type I , Chemistry , Genetics , Metabolism , Sequence Homology, Amino Acid摘要
<p><b>OBJECTIVE</b>To investigate the frequencies of -1470, -511 and -31 single nucleotide polymorphisms (SNPs) in the promoter of IL-1beta and its haplotype constitution in Chongqing population.</p><p><b>METHODS</b>One hundred and twelve healthy Chongqing people were enrolled in this study. Polymorphisms at -1470 (G to C), -511 (T to C) and -31 (C to T) of IL-1beta were genotyped with the method of restriction fragment length polymorphism (RFLP). Haplotype frequencies were analyzed by Arlequine software.</p><p><b>RESULTS</b>Frequencies of IL-1beta -1470, -511 and -31 SNPs were 41.67%, 50% and 45.33%, respectively. Genotype frequencies of -1470 locus were 39.81%, 37.04% and 23.15% for G/G, G/C and C/C respectively. As for T-511C SNP, genotype frequencies of T/T, T/C and C/C were 29.91%, 40.18% and 29.91%, respectively. Genotyping results of C/C, C/T, and T/T of -31 locus were 35.51%, 38.32% and 26.71% respectively. Haplotype analysis found that there were mainly three haplotypes constituted by three SNPs, ie., G-T-C, C-T-C and G-C-T.</p><p><b>CONCLUSIONS</b>Polymorphisms exist in the promoter of IL-1beta in Chongqing population. Three SNPs locate in the same haplotype block.</p>
Subject(s)
Female , Humans , Male , Asian People , Genetics , China , Cohort Studies , Genetics, Population , Genotype , Haplotypes , Interleukin-1 , Genetics , Polymerase Chain Reaction , Polymorphism, Restriction Fragment Length , Polymorphism, Single-Stranded Conformational , Promoter Regions, Genetic , Sensitivity and Specificity摘要
<p><b>OBJECTIVE</b>Early gestational mammalian fetuses possess the amazing ability to heal cutaneous wounds in a scarless fashion. Over the past years, scientists have been working to decipher the mechanisms underlying this regenerative repair. The remarkable phenotypic differences between fetal and adult healings behooves us to learn their characteristics in genetics, which represents potentially important mechanisms involved in wound repair observed in fetal versus adult tissues. In this sense, it is reasonable to construct subtractive cDNA library for future research.</p><p><b>METHODS</b>Middle laparotomy and hysterotomy were performed on pregnant rabbits at 20-day gestation to expose the fetal back, and a longitudinal incision through the skin was made on the back of the fetus. The traumatized fetal skin was harvested 12 hours post-operation, the fetus control and traumatized adult skin specimens were taken at the same time. dscDNA was synthesized from total RNA of skin samples with SMART technology. Taking one of the three samples as Tester respectively and the other two as Drivers, we obtained 1 forward and 2 reverse hybridization products. After being amplified with selective polymerase chain reaction, the products were inserted into a vector, and then transferred into E. coli HB101. The colonies were screened afterwards.</p><p><b>RESULTS</b>The wounded fetuses were alive for a long time even after birth. Every determinant step, such as RNA isolation, cDNA synthesis, Rsa I digestion, adaptor ligation and hybridization, was well-operated. Subtractive efficiency identification demonstrated that the suppression subtractive hybridization (SSH) was successful. Insertion into vector and transferring to E. coli were satisfactory.</p><p><b>CONCLUSIONS</b>Instead of classic SSH, an improved SSH with 2 Drivers was applied for the experiment. Results confirmed that the improved program was reasonable and correct in both theory and practice. The subtractive cDNA library we have obtained is going to be used for future researches to reveal scarless healing related gene(s) and its (their) expression.</p>
Subject(s)
Animals , Female , Pregnancy , Rabbits , Cicatrix , Genetics , DNA, Complementary , Disease Models, Animal , Fetus , Gene Library , Polymerase Chain Reaction , Pregnancy, Animal , RNA, Messenger , Sensitivity and Specificity , Skin , Wounds and Injuries , Wound Healing , Genetics , Physiology摘要
Objective To investigate the relationship between CIDE-B change and apoptosis by detecting expression of CIDE-B gene and protein under various injuring conditions(firearm injury and in- cision injury)and comparing neuronal apoptosis.Methods A total of 108 New Zealand rabbits were randomized into firearm injury group,incision injury group and control group.Samples were harvested at different time points after injury for measuring CIDE-B mRNA expression level by RT-PCR,demonstrating morphological distribution of CIDE-B mRNA by in situ hybridization and determining CIDE-B protein level by Western blot.Results Following nerve injury,CIDE-B mRNA expression in spinal cord was en- hanced markedly in the firearm injury group,increased from at day 1,reached peak at day 7,lasted for two weeks and declined after four weeks.CIDE-B mRNA expression was increased late,with small range in the incision injury group.The changes in CIDE-B protein level accorded with those in CIDE-B mRNA level.There was slight CIDE-B mRNA expression,distributing in spinal cord neuronal cytoplasma,in spinal cord tissues in the control group.Conclusions Following peripheral nerve injury,expressions of CIDE-B mRNA and protein are enhanced in spinal cord neurons and distributed in the grey matter of spi- nal cord.Such enhancement is more marked after firearm injury than incision injury.