ABSTRACT
Cell identification and counting in living and coculture systems are crucial in cell interaction studies, but current methods primarily rely on complicated and time-consuming staining techniques. Here, a label-free method to precisely recognize, identify, and instantly count cells in situ in coculture systems via combinational machine learning models s presented. A convolutional neural network (CNN) model is first used to generate virtual images of cell nuclei based on unlabeled phase-contrast images. Coordinates of all the cells are then returned according to the virtual nucleus images using two clustering algorithms. Finally, phase-contrast images of single cells are cropped based on the coordinates and sent into another CNN model for cell-type identification. This combinational approach is highly automatic and efficient, which requires few to no manual annotations of images in the training phase. It shows practical performance in different cell culture conditions including cell ratios, densities, and substrate materials, having great potential in real-time cell tracking and analyzing.
Subject(s)
Cell Nucleus/metabolism , Image Processing, Computer-Assisted/methods , Algorithms , Cells, Cultured , Cluster Analysis , Human Umbilical Vein Endothelial Cells , Humans , Machine Learning , Neural Networks, Computer , Single-Cell AnalysisABSTRACT
Generating a biofunctional film that can mimic the extracellular matrix (ECM) in an efficient and robust technique that may have great potential for medical devices, tissue engineering, and regenerative medicines. Herein, a facile approach to generate ECM biomimetic films based on the humidity-triggered relaxation of polyelectrolyte complex (PEC) nanoparticles is reported. The poly(l-lysine) and hyaluronan are precomplexed and sprayed onto a substrate, which, via a trigger of vaporous water, can be transformed into an even and stable film. The spontaneous polymer chain interfusion (diffusion coefficient ≈1.01 × 10-9 cm2 s-1 ) under saturated humidity, allowing for the rapid reorganization (within 30 min) of film morphology and structure is demonstrated. A controllable and scalable way for the loading of diversified bioactive agents, as well as on-demand modulation of stiffness is further presented. Moreover, the high-throughput arrays and programmed patterns can be easily completed, suggesting huge potentials that surpass those of state-of-the-art methods. Combined with high efficiency and flexible functionalization, it is believed that this approach should be beneficial for extending the practical applications of PEC films, such as medical implants, chip detectors, and so on.
Subject(s)
Biomimetics , Hyaluronic Acid , Extracellular Matrix , Humidity , PolyelectrolytesABSTRACT
Phosphorylcholine (PC) based polymer coatings with excellent biocompatibility have shown successful commercialization in drug-eluting stents. However, poor degradability represents a challenge in the application of biodegradable stents. Herein, a biodegradable phosphorylcholine copolymer is developed based on one-step radical ring-opening polymerization (RROP). This copolymer was synthesized by copolymerization of a PC unit, degradable ester (2-methylene-1,3-dioxepane, MDO) unit and non-degradable butyl methacrylate (BMA) unit, which showed ratio controllability by changing the monomer ratio during polymerization. We demonstrated that the copolymer with the ratio of 34% MDO, 19% MPC and 47% BMA could form a stable coating by ultrasonic spray, and showed good blood compatibility, anti-adhesion properties, biodegradability, and rapamycin eluting capacity. In vivo study revealed its promising application as a biodegradable stent coating. This work provides a facile path to add biodegradability into PC based polymers for further bio-applications.
Subject(s)
Cardiovascular Diseases/surgery , Coated Materials, Biocompatible/chemistry , Phosphorylcholine/chemistry , Polymers/chemistry , Stents , Animals , Cardiovascular Diseases/metabolism , Cardiovascular Diseases/pathology , Cells, Cultured , Coated Materials, Biocompatible/chemical synthesis , Coated Materials, Biocompatible/metabolism , Molecular Structure , Particle Size , Phosphorylcholine/chemical synthesis , Phosphorylcholine/metabolism , Polymers/chemical synthesis , Polymers/metabolism , Rabbits , Surface Properties , Swine , Swine, MiniatureABSTRACT
In the "post-antibiotic era", healthcare-associated infection has become a global problem that threatens public health and causes huge economic losses. The development of antibacterial coatings based on non-antibiotic strategies is particularly important as drug-resistant bacteria continue to evolve. Photodynamic coatings are a high potential method to treat bacteria, however, the aggregation of photosensitizers on the coating affects the photodynamic capacity seriously. Herein, a photodynamic coating is developed based on the host-guest interaction between ß-cyclodextrin and the photosensitizer methylene blue (MB). The host-guest interaction avoids aggregation of MB and results in a high singlet oxygen quantum yield. Consequently, efficient photoantibacterial activity towards methicillin-resistant Staphylococcus aureus is achieved by the photodynamic coating with very low MB density (0.53 ± 0.06 µg cm-2).
Subject(s)
Coated Materials, Biocompatible/chemistry , Methylene Blue/chemistry , Photosensitizing Agents/chemistry , Coated Materials, Biocompatible/chemical synthesis , Drug Liberation , Humans , Hydrophobic and Hydrophilic Interactions , Methicillin-Resistant Staphylococcus aureus/drug effects , Methylene Blue/metabolism , Methylene Blue/pharmacology , Photosensitizing Agents/metabolism , Photosensitizing Agents/pharmacology , Quantum Theory , Singlet Oxygen/metabolism , Static Electricity , Ultraviolet Rays , beta-Cyclodextrins/chemistryABSTRACT
Medical coatings cooperated with biomacromolecules can regulate biological events and tissue responses, thus increasing medical implant longevity and providing improved and/or new therapeutic functions. In particular, medical coatings, which can load the correct species and doses of biomacromolecules according to individual diagnoses, will significantly optimize treatment effects and satisfy the rising clinical need of "precision medicine". Herein, we report on a dynamic microporous coating with an ultrafast self-healing property to fulfill the "load-and-play" concept for "precision medicine". A structure-switchable coating based on poly(ε-caprolactone)-poly(ethylene glycol)-poly(ε-caprolactone) triblock copolymer network is constructed. The coating can be switched to microporous morphology via a water swelling and freeze-drying process. Then, through a mild thermo-trigger as low as 40⯰C, this spongy coating can undergo self-healing to switch back to a pore-free structure within minutes to even 5â¯s. Based on this dynamic coating, we suggest a simple and versatile method to encapsulate biomacromolecules for surface-mediated delivery. The ultrafast self-healing of the microporous structure enables uniform incorporation of biomacromolecules with an easily achieved high loading of albumin of 16.3⯵g/cm2 within 1â¯min. More importantly, controllable encapsulation can be realized by simple control of the concentration of the loading solution. We further demonstrate that the encapsulated biomacromolecules retained their bioactivity. This work may benefit clinicians with flexibility to provide personalized medical coatings for individual patients during treatment.
Subject(s)
Delayed-Action Preparations/chemistry , Polyesters/chemistry , Polyethylene Glycols/chemistry , Animals , Drug-Eluting Stents , Human Umbilical Vein Endothelial Cells , Humans , Porosity , Rabbits , Serum Albumin, Bovine/administration & dosage , Serum Albumin, Bovine/pharmacokinetics , Temperature , Vascular Endothelial Growth Factor A/administration & dosage , Vascular Endothelial Growth Factor A/pharmacokineticsABSTRACT
<p><b>OBJECTIVE</b>To investigate the relationship between epidermal growth factor (EGF) gene polymorphisms at G-61A, R431K, and D784V and susceptibility to silicosis.</p><p><b>METHODS</b>In a case-control study, 116 patients diagnosed with stage I silicosis were included in the case group, and 149 workers without silicosis of the same gender and nationality, exposed to the same nature of dust, and with similar age and cumulative time of dust exposure were included in the control group. Peripheral venous blood was collected, DNA was extracted by salting out, polymerase chain reaction-restriction fragment length polymorphism was used to identify the genotypes at three polymorphic loci of EGF and the allele frequencies, and their distributions in the case group and control group were analyzed.</p><p><b>RESULTS</b>The genotype frequencies of G-61A GG, GA, and AA in the case group were 50.9%, 34.5%, and 14.7%, respectively, and significant differences were found when comparing the data with those in the control group (35.6%,44.3%, and 20.1%), (χ(2) = 6.283, P = 0.048). The distribution frequencies of allele A in the case group and control group were 31.9%and 42.3%, respectively, and the difference between the two groups was statistically significant (χ2 = 5.554, P = 0.018). The risk of silicosis in workers carrying allele G at G-61A was increased by 1.564 times (OR = 1.564, 95%CI: 1.092∼2.024). The genotype frequencies of D784V AA, AT, and TT in the case group were 58.6%, 34.5%, and 6.9%, respectively, versus 65.1%, 31.5%, and 3.4% in the control group, and the differences between the two groups were not statistically significant (χ(2) = 2.278, P = 0.320). The genotype frequencies of R431K GG, GA, and AA in the case group were 56.9%, 39.7%, and 3.4%, respectively, versus 55.0%, 39.6%, and 5.4% in the control group, and the differences between the two groups were not statistically significant (χ(2) = 0.572, P = 0.751).</p><p><b>CONCLUSION</b>The EGF gene polymorphism at G-61A is associated with susceptibility to silicosis, and the risk of silicosis in dust-exposed workers carrying GG genotype is relatively high. No relationship between EGF gene polymorphisms at D784V and R431K and silicosis is found.</p>
Subject(s)
Aged , Aged, 80 and over , Humans , Middle Aged , Alleles , Case-Control Studies , Epidermal Growth Factor , Genetics , Gene Frequency , Genetic Predisposition to Disease , Genotype , Polymorphism, Single Nucleotide , Silicosis , Epidemiology , GeneticsABSTRACT
<p><b>OBJECTIVE</b>To investigate the autophagy of effector cells in lung tissue at different time points when rats were exposed to free SiO2 dust.</p><p><b>METHODS</b>Sixty Wistar rats (220∼230 g) were selected and allocated to experimental group (n = 30) and control group (n = 30). In the experimental group, a rat silicosis model was established by infusing SiO2 suspension into the trachea of rats. Six rats in each group were sacrificed on days 1, 7, 14, 21, or 28 of dust exposure. Lung tissue samples were collected to prepare lung tissue sections. The pulmonary inflammation and fibrosis were observed by HE staining. The proautophagosome, autophagosome, and autophagolysosome in lung tissue sections were observed under a transmission electron microscope.</p><p><b>RESULTS</b>On day 1 of dust exposure, many proautophagosomes and autophagosomes were seen in both experimental group and control group. On day 7 of dust exposure, the experimental group had more autophagosomes in lung tissue than the control group. On day 14 of dust exposure, the experimental group had fewer autophagosomes than the control group. On days 21 and 28, autophagolysosomes were seen in macrophage plasma in both experimental group and control group; the autophagolysosomes in experimental group showed cloudy swelling and expansion, and some were vacuolated, and these changes were more significant on day 28.</p><p><b>CONCLUSION</b>Free SiO2 dust can induce autophagy in the lung tissue of rats, with varying degrees at different time points of dust exposure.</p>
Subject(s)
Animals , Male , Rats , Autophagy , Dust , Lung , Pathology , Rats, Wistar , Silicon Dioxide , ToxicityABSTRACT
<p><b>OBJECTIVE</b>The purposes of this thesis were to study the behavior about workers exposed to dust and provide scientific basis for health promotion.</p><p><b>METHODS</b>We designed a questionnaire and carry it on the 746 dust workers in the 3 representative corporations of Machinery, Ceramic, and Metallurgy Industry. All data were input into computer. And a database was established with Excel. SPSS11.5 statistical analysis software was used to analyze the influence on protecting behavioral between the application of qualifications, different jobs, training or protection, and other aspects etc.</p><p><b>RESULTS</b>The rates were 94.4% and 75.3% about the regular physical examination and requirements for protective equipment. The rate of choosing an effective way of protection was generally low (15.4%). There was significant difference for among different educational background workers (P < 0.01). The rates of choosing an effective way of protection (20.3%), the regular physical examination (98.3%) and requirements for protective equipment (86.4%) in the dust workers who participated in the training of dust protection were superior than those who did not participated in the training. There was the significant difference (P < 0.05, P < 0.01). There was the significant difference for the rate of effective way of protection, regular physical examination, and requirements for protective equipment among the different corporations (P < 0.05).</p><p><b>CONCLUSIONS</b>Dust workers' using rate about the choosing an effective way of protection was generally low in Machinery, Ceramic, and Metallurgy Industry. Those who were not educated had a lower using rate about the protection behavior, regular physical examination, and requirements for protective equipment than those educated.</p>
Subject(s)
Adolescent , Adult , Female , Humans , Male , Middle Aged , Young Adult , Ceramics , Choice Behavior , Dust , Industry , Metallurgy , Occupational Exposure , Respiratory Protective DevicesABSTRACT
<p><b>OBJECTIVE</b>To study the roles of macrophage apoptosis, IL-1, and IL-8 in the pathogenesis of rat pulmonary fibrosis induced by silica.</p><p><b>METHODS</b>Forty eight male Wistar rats were divided into the 4 control groups (24 rats) and 4 experimental groups (24 rats). Rats in the control groups were treated with 1 ml normal saline by trachea instillation, whereas the rats in experimental groups were exposed 1 ml silica suspension (100 mg/ml) by trachea instillation for 1, 7, 14 and 28 days, respectively. Six rats of each group were sacrificed, then the bronchoalveolar lavage fluid and lung tissues were collected, respectively. Pulmonary inflammation, fibrosis and other pathological changes were detected with H.E. staining. Morphological changes of the early stage apoptosis in macrophages were detected with transmission electron microscope (TEM). The early apoptosis rates of macrophages in BALF were also assessed using Annexin V-FITC/PI kit. The IL-1 and IL-8 levels of serum were measured with the ELISA.</p><p><b>RESULTS</b>The apoptotic rates (11.48% +/- 0.24%, 16.03% +/- 0.68%, 15.53% +/- 1.07%, 18.92% +/- 2.70%, respectively) of macrophage in the experimental groups increased obviously with time, as compared to the controls (5.47% +/- 2.06%, 6.39% +/- 0.215, 9.07% +/- 0.61% and 8.54% +/- 0.16%, Respectively) (P < 0.05). The IL-1 levels of serum in the experimental groups were 23.64 +/- 0.84, 23.38 +/- 1.10, 22.21 +/- 0.86 and 24.29 +/- 1.31 pg/ml, respectively, which were significantly higher than those (18.52 +/- 1.23, 18.40 +/- 1.6, 17.92 +/- 2.21 and 18.53 +/- 2.64 pg/ml, respectively) in the control groups (P < 0.05) without time-effect relationship. The serum IL-8 levels on the 1st, 7th and 14th days in the experimental groups were 21.32 +/- 1.44, 21.90 +/- 2.08 and 22.00 +/- 2.80 pg/ml, respectively, which were significantly higher than those (17.69 +/- 1.09, 16.98 +/- 2.09 and 17.54 +/- 1.62 pg/ml, respectively) in the control groups (P < 0.05).</p><p><b>CONCLUSION</b>The early macrophage apoptosis and changes of IL-1 and IL-8 may in lungs may play an important role in the development of pulmonary fibrosis induced by silica.</p>
Subject(s)
Animals , Male , Rats , Apoptosis , Disease Models, Animal , Interleukin-1 , Blood , Interleukin-8 , Blood , Macrophages, Alveolar , Cell Biology , Pulmonary Fibrosis , Rats, Wistar , Silicon Dioxide , Toxicity , Silicosis , Blood , PathologyABSTRACT
<p><b>OBJECTIVE</b>To explore the relationship between polymorphisms of interleukin-4 (IL-4) gene (-33, +45, VNTR, +429, +448) and the susceptibility of silicosis.</p><p><b>METHODS</b>In a case-control study, the case group consisted of 101 patients with silicosis, and was matched with the control group (121 workers without silicosis), according to the age, sex, nationality, working place, exposure to dust. The polymorphisms of IL-4 (five locus) detected by the method of polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) techniques.</p><p><b>RESULTS</b>There was no difference of age, exposure and smoking between case group and control group (P > 0.05). The two groups had good comparability. Only the GA genotype in the IL-4 (+429) locus was found, the genotypes of AA and GG were not found. The CC genotype in the IL-4 (+448) locus was found, the genotypes of CG and GG were not found. The frequencies of AA, GG and AG of IL-4 (+45) locus in the case and control groups were 55.4%, 10.9%, 33.7% and 62.0%, 11.6%, 26.4%, respectively, there was no the significant difference between case and control groups (P > 0.05). The frequencies of B1B1, B2B2 and B1B2 of IL-4 (VNTR) locus in the case and control groups were 73.3%, 1.0%, 25.7% and 68.6%, 1.7%, 29.8%, respectively, there was no the significant difference between case and control groups (P > 0.05). The frequencies of TT, CC and CT in IL-4 -33 locus in the case group were 55.4%, 11.9% and 32.7%, which were significantly higher than those (69.4%, 4.1%, 26.4%) in control group (P < 0.05).</p><p><b>CONCLUSIONS</b>There was no relationship between IL-4 (+45, VNTR) genotypes and prevalence of silicosis in this study. The polymorphisms of IL-4 (+448) site were not found which may be related to the race. The relationship between genetic polymorphism of IL-4 (-33) locus and silicosis development was found, Workers with IL-4 (-33) allele C are susceptible to the silica.</p>
Subject(s)
Aged , Aged, 80 and over , Humans , Male , Middle Aged , Alleles , Asian People , Genetics , Case-Control Studies , Gene Frequency , Genetic Predisposition to Disease , Genotype , Interleukin-4 , Genetics , Polymorphism, Single Nucleotide , Silicosis , Epidemiology , GeneticsABSTRACT
<p><b>OBJECTIVE</b>To explore the relationship between polymorphisms of interleukin-4 (IL-4) gene (-33, +45, intron3, +429, +448) and the susceptibility of silicosis.</p><p><b>METHODS</b>A case-control study was carried out. 101 silicosis patients were selected as cases. As strictly matching, 121 of non silicosis workers were selected as the controls. The polymophisms of IL-4 (five locus) were detected by the method of polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) techniques.</p><p><b>RESULTS</b>The GA genotype in the IL-4+429 locus and the CC genotype in the IL-4+448 locus were found. The frequencies of AA, GG and AG of IL-4+45 locus in the cases were 55.4%, 10.9%, and 33.7% and in the controls were 62.0%, 11.6%, and 26.4%. The differences between cases and controls were not significant. The frequencies of B1B1, B2B2, and B1B2 of intron3 VNTR locus in the cases were 73.3%, 1.0%, and 25.7% and in the controls were 68.6%, 1.7%, and 29.8%. The differences were not significant. The frequencies of TT, CC, and CT in -33 locus in the cases were 55.4%, 11.9%, and 32.7% and in the controls were 69.4%, 4.1%, and 26.4%. The differences were significant (P=0.034).</p><p><b>CONCLUSION</b>The relationship between genetic polymorphism of IL-4-33 site and silicosis has been found and -33TT is a protective genotype for silicosis.</p>
Subject(s)
Aged , Aged, 80 and over , Humans , Middle Aged , Case-Control Studies , China , Epidemiology , Data Interpretation, Statistical , Gene Frequency , Genetic Predisposition to Disease , Genotype , Gold , Interleukin-4 , Genetics , Mining , Polymerase Chain Reaction , Polymorphism, Restriction Fragment Length , Silicosis , Epidemiology , GeneticsABSTRACT
<p><b>OBJECTIVE</b>To investigate the effect of occupational stress on menses and sex hormones.</p><p><b>METHODS</b>415 female knitting workers were investigated using the generic job stress questionnaire. Their venous blood were collected and the six sex hormones were detected by using radio-immune method. The different rate of abnormal menses and sex hormones level between different stress degree groups were analyzed.</p><p><b>RESULTS</b>The abnormal rate of menses, menstrual blood volume, menstrual cycle, menstrual period was 36.24%, 19.80%, 14.43%, 11.41% respectively. The prevalence rate of dysmenorrheal and premenstrual syndrome was 1.01% and 29.19% respectively. The more depression, the higher menses disorders in non-intrauterine device (IUD) group. The more job demands, the higher daily stress in IUD group while the longer work time, the more abnormal menstrual period in two groups. More physical symptoms and deeper depression in non-IUD group were related to higher abnormal rate of menstrual blood volume. The level of blood E2 was lower in the group of prolonged work-time than that of in normal work-time group. The increasing FSH level and decreasing T level was associated with higher job demands. Multiple factor analysis showed that physical symptom, control of resource and negative life affairs were the risk factors of menses disorder; The physical symptom was the risk factor of menstrual blood volume; More physical symptoms, less positive feeling and shift were the risk factors of premenstrual syndrome; Less positive feeling was the risk factor of menstrual cycle; Prolonged daily work-time was the risk factor of menstrual period.</p><p><b>CONCLUSION</b>Higher stress degree can lead to higher FSH and E2 and lower T level,and induce menses disorder.</p>
Subject(s)
Adult , Female , Humans , Analysis of Variance , Burnout, Professional , Chi-Square Distribution , Gonadal Steroid Hormones , Blood , Logistic Models , Menstruation , Physiology , Surveys and Questionnaires , Textile IndustryABSTRACT
<p><b>OBJECTIVE</b>To explore the relationship between the genetic polymorphism of fibronectin (FN) (4 genetic locus) and pneumoconiosis.</p><p><b>METHODS</b>128 male I-period pneumoconiosis were selected as cases who were examined with radiography and diagnosed by the Pneumoconiosis Diagnosis Expert Panel, based on the Chinese National Diagnosis Criteria of Pneumoconiosis (GBZ 70 - 2002). According to 1:1 paired matching method, 128 dust exposure workers were selected as control who were exposed to same dust as cases. The difference of age and cumulative length of service between case and control was not over five years and two years, respectively. 5 ml peripheral venous blood was drawn and anticoagulated with 2% EDTA. The polymorphisms of FN (MspI, TaqIb, HindIII, HaeIIIb) were detected, using the method of polymerase chain restriction fragment length polymorphism (PCR-RFLP) techniques and PCR.</p><p><b>RESULTS</b>The frequencies of FN Msp I (CC) in cases and control groups were 10.9% and 3.9%, respectively. The difference was significant (P < 0.05). The frequencies of FN (MspI) C allele were 41.8% and 31.2% in case and control, and the difference between cases and controls was significant (P < 0.05). The frequencies of FN HaeIIIb (AA) genotype in cases (24.2%) was higher than that in control groups (17.9%), OR = 5.0 (95% CI: 4.840 approximately 24.210). The frequencies of FN (HaeIIIb) A allele were 51.9% and 42.2% in case and control, and the difference was significant (P < 0.05). The difference of TaqIb and HindIII genotype between cases and controls were not significant (P > 0.05).</p><p><b>CONCLUSION</b>The risk of suffering from pneumoconiosis increases in workers carrying FN (MspICC or HaeIIIb AA) genotype after exposure to dust. Workers both carrying FN (HaeIIIb AA) and (MspICC) genotypes are more susceptible to pneumoconiosis. The relationship between genetic polymorphism of FN (TaqIIb, HindIII) and pneumoconiosis has not been found.</p>
Subject(s)
Adult , Aged , Aged, 80 and over , Humans , Male , Middle Aged , Fibronectins , Genetics , Genetic Predisposition to Disease , Genotype , Pneumoconiosis , Genetics , Polymorphism, Restriction Fragment LengthABSTRACT
<p><b>OBJECTIVE</b>To investigate the occupational stressors and modifiers of pediatricians and nurses in order to find the measurements for control of the job stress.</p><p><b>METHODS</b>427 pediatricians and nurses working in five hospitals of a city served as subjects. Of them, the staff in section of pharmacy and toll offices in each hospital mentioned above served as control group. The General Job Stress Questionnaire was used to investigate the job stress by self-assessment.</p><p><b>RESULTS</b>The scores of job demand, job risk, drug using, daily job stress, positive feelings, patient A behavior, physical environment and feeling balance in pediatricians and nurses were higher than those of control group, but the scores of job-person conflict, environmental control, technology utility, mental health, responsibility on things were lower than those of control group (P<0.05). The points of job future, job locus of control, self-esteem, job satisfaction, job load variance, depression in nurses were higher than those of pediatricians, and non-work activities, job risk and daily life stress were lower than those of doctors (P<0.05). The main affecting factors on job strain of pediatric staff included job monotony, higher job demand, more non-work job, lower job control, more job risk, job future ambiguous, poorer social support, lower job locus control and lower self-esteem.</p><p><b>CONCLUSION</b>The stress degree of pediatric staff is higher than that of controls. The pediatricians have more job stress than that of nurses. The main stressors of pediatric staff are job monotony, higher job demand, more non-worker activity, lower job control, higher job risk and ambiguous job future. The main modifiers are good social support, external job locus of control and higher self-esteem.</p>
Subject(s)
Adult , Female , Humans , Male , Young Adult , Burnout, Professional , Medical Staff, Hospital , Psychology , Nursing Staff, Hospital , Psychology , Pediatrics , Surveys and QuestionnairesABSTRACT
<p><b>OBJECTIVE</b>To study the expression level of cyclin D1-CDK4 protein in human embryonic lung fibroblasts (HELF) induced by quartz, and to study whether the expression level of cyclin D1-CDK4 protein mediated by mitogen activated protein kinase (MAPK)/(AP-1) signaling pathways.</p><p><b>METHODS</b>Cells were harvested after stimulation 2 h for the detection of cytokines. Cyclin D1 and CDK4 (cyclin dependent kinase 4) proteins was measured by immunocytochemistry (IC) and Western blot (WB).</p><p><b>RESULTS</b>The exposure of HELF to crystalline quartz for 2 hours could cause the decrease of cyclin D1 and cyclin dependent kinase 4 (CDK4) protein expression level, (7.91 +/- 0.29) x 10(3) and (5.17 +/- 0.28) x 10(4) respectively, which was lower than that of the HELF group (P < 0.05). AG126 (chemical inhibitor of the extracellular signal regulated protein kinase (ERK) signaling pathway) and the dominant negative mutant of ERK2 (molecular inhibitor of ERK2), prevented the decrease of cyclin D1 and CDK4 protein expression level. The chemical inhibitor of c-Jun NH2-terminal amino kinase (JNK), SP600125, could prevent both cyclin D1 and CDK4 protein expression level decrease. But SB203580, the chemical inhibitor of p38, prevented neither cyclin D1 nor CDK4 protein expression level decrease. Curcumin could prevent CDK4 protein expression level decrease but not cyclin D1 protein.</p><p><b>CONCLUSION</b>ERKs and JNKs, but not p38, are responsible for the decrease of cyclin D1 and CDK4 protein expression level in HELF induced by quartz. AP-1 is responsible for the decrease of CDK4 expression level but not that of cyclin D1.</p>
Subject(s)
Humans , Cells, Cultured , Cyclin D1 , Metabolism , Cyclin-Dependent Kinase 4 , Metabolism , Fibroblasts , Metabolism , Lung , Cell Biology , Embryology , Mitogen-Activated Protein Kinases , Metabolism , Quartz , Toxicity , Transcription Factor AP-1 , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To explore association genetic polymorphism of XPD with chromosomal damage in workers exposed to radiation.</p><p><b>METHODS</b>182 workers exposed to radiation for at least one year with chromosomal damage were selected as cases based on a general health examination for all workers exposed to radiation in Tangshan city. The control group without chromosomal damage was matched to case by age (within 5 years), sex, work unit, type of exposed to radiation, cumulate serve length (within 1 year) according to 1:1. The micro whole blood cultivation was used for the chromosome analysis. The chromosome aberration type and rate were observed and counted. The polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was used to examine the genotype of three XPD loci (751, 312 and 156).</p><p><b>RESULTS</b>The frequency of XPD 751 AA in cases was higher than that in controls (P < 0.05). The frequency of 751 allele in case group was statistically higher than that in the control groups (P < 0.05). No statistical difference was found in the frequencies of XPD 312 genotype and allele between the case and control group (P > 0.05). 156 mutant gene type in case group was higher than that in the control groups. The frequency of 156 A allele in case group were higher than that of the control groups (P < 0.05). The frequency of genotype with both 751AA and 156CA or 751AA and 156AA was higher in cases than that of controls (P < 0.05).</p><p><b>CONCLUSION</b>XPD 751AA genotype is a possible risk factor for radiation-induced chromosomal damage. XPD 156 mutant gene type is a possible risk factor for radiation-induced chromosomal damage. Individuals with both XPD 751AA and 156 (CA+AA) genotypes are susceptible to radiation-induced chromosomal damage. No association of XPD 312 polymorphism with radiation-induced chromosomal damage is found.</p>
Subject(s)
Adult , Female , Humans , Male , Middle Aged , Young Adult , Case-Control Studies , Chromosome Aberrations , Radiation Effects , Genetic Predisposition to Disease , Occupational Exposure , Polymorphism, Restriction Fragment Length , Radiation , Xeroderma Pigmentosum Group D Protein , GeneticsABSTRACT
<p><b>OBJECTIVES</b>To study the phosphorylation level of mitogen activated protein kinase (MAPK) in human embryonic lung fibroblasts (HELF), and the expression level of cyclin D1-CDK4 protein in S-HELF and whether the expression level of cyclin D1-CDK4 protein mediated by MAPK/AP-1 signaling pathway in S-HELF.</p><p><b>METHODS</b>Two kinds of treatment: (1) Cells were harvested after stimulation 2 h for the detection of cytokines. (2) Cells were stimulated by quartz for a long time (2 months) for transformation characters (S-HELF). The MAP kinase was detected by western blot. Cyclin D1 and CDK4 (cyclin dependent kinase 4) proteins was measured by immunocytochemistry. Flow cytometry was used to evaluate the alternation of cell cycle.</p><p><b>RESULTS</b>Crystalline quartz could cause the phosphorylation level of ERKs, p38K, and JNKs in HELF increase. However, activated levels of ERKs and p46 of JNKs increased in S-HELF, and p38K activation decreased, and no effect on activation of p54 of JNKs, as compared with those in parental HELF. Cyclin D1 and CDK4 protein expression levels increased in S-HELF as compared with parental HELF. Inhibition of ERKs activation by AG126, AP-1 by curcumin, and JNKs by SP600125 could reduced the induction of cyclin D1 and CDK4, whereas inhibition of p38K by SB203580 did not show any inhibitory effects on S-HELF.</p><p><b>CONCLUSIONS</b>The phosphorylation levels of ERK1/2, JNK1/2, and p38 increased in HELF exposed to quartz. The phosphorylation levels of ERK1/2 and JNK1 increased, but the phosphorylation level of p38 decreased in S-HELF. The expression level of cyclin D1-CDK4 protein increased in S-HELF. Overexpression of cyclin D1-CDK4 is due to the activation of ERKs, JNKs/AP-1 signaling pathway in S-HELF.</p>
Subject(s)
Humans , Cell Line , Cyclin D1 , Metabolism , Cyclin-Dependent Kinase 4 , Metabolism , Fibroblasts , Cell Biology , Metabolism , Lung , Cell Biology , MAP Kinase Signaling System , Mitogen-Activated Protein Kinases , Metabolism , Quartz , ToxicityABSTRACT
<p><b>OBJECTIVE</b>To explore the association between genetic polymorphisms of TGF-beta (TGF-beta) and susceptibility to pneumoconiosis.</p><p><b>METHODS</b>One hundred and seventeen patients with pneumoconiosis were selected as case. The control group was workers exposed to dust but without pneumoconiosis who had the same sex, nationality, and workshop or work site as case. The differences in the age and cumulative exposure time between the case and control group were not move than five years and two years, respectively. The case matched with the control according to 1:1. Polymerase chains reaction-restriction fragment length polymorphism was used to determine the frequencies of TGF-beta genes in the two groups.</p><p><b>RESULTS</b>The frequencies of this TGF-beta (-509) genotypes were CC (22.2%), CT (43.6%) and TT (34.2%) in cases, which was significantly different from the control group, respectively (OR = 1.390, P < 0.05). There was no significant difference for frequency of TGF-beta+869 genotypes and allelic between case and control (P > 0.05). The frequencies of the TGF-beta (+915) genotypes in case [GG (70.9%), GC (29.1%)] were significantly different from the control group (OR = 1.455, P < 0.05). The frequency of TGF-beta (+915) * C allele in the case and control was 14.5% and 8.5%, respectively (P < 0.05). The frequencies of carrying TGF-beta (-509) CC and (+915) GG genotypes were 12.8% and 29.9% in case and control. The frequencies of carrying TGF-beta (-509) * T and (+915) * C alleles were 9.8% and 5.1% in pneumoconiosis and control (P < 0.05).</p><p><b>CONCLUSIONS</b>TGF-beta (-509)CC genotype may be the protective factor for the pneumoconiosis. TGF-beta (+915)GC genotype may be a susceptible factor for the pneumoconiosis. The workers of carrying TGF-beta (-509) * T and (+915) * C alleles are more susceptible to pneumoconiosis.</p>
Subject(s)
Adult , Aged , Aged, 80 and over , Humans , Male , Middle Aged , Case-Control Studies , Gene Frequency , Genotype , Pneumoconiosis , Genetics , Polymorphism, Single Nucleotide , Transforming Growth Factor beta1 , GeneticsABSTRACT
<p><b>OBJECTIVE</b>To explore the relationship between interleukin-6 (IL-6) (-634C/G) genetic polymorphisms and the pneumoconiosis.</p><p><b>METHODS</b>A total of 104 male stage I pneumoconiosis patients diagnosed by the Pneumoconiosis Diagnosis Expert Panel according to the Chinese National Diagnosis Criteria of Pneumoconiosis (GBZ 70 - 2002) were selected. The pneumoconiosis comprised 66 silicosis and 38 coal worker' pneumoconiosis (CWP). A total of 122 workers exposed to same dusts as the patients but without pneumoconiosis including 77 exposed to silica dusts and 45 to coal dusts were selected. The patients and the controls had the same dust exposure history. The peripheral venous blood was drawn from each subject. The IL-6 (-634C/G) genetic polymorphisms were detected by polymerase chain reaction restriction fragment length polymorphisms (PCR-RFLP) techniques.</p><p><b>RESULTS</b>The frequencies of IL-6 (-634C/G) (CC, CG and GG) genotypes were 66.7%, 19.7% and 13.6% in silicosis group, 42.9%, 42.9% and 14.2% in silica dust exposure group, 73.7%, 18.4% and 7.9% in CWP group, 51.1%, 35.6% and 13.3% in coal dust exposure group respectively. The statistical analysis indicated that there was significant difference in the frequencies of IL-6 (-634C/G) (CC, CG and GG) genotypes between silicosis patients and workers exposed to silica dusts (P < 0.05).</p><p><b>CONCLUSION</b>IL-6 (-634 C/G) genetic polymorphisms might play a role in the occurrence of silicosis.</p>
Subject(s)
Adult , Aged , Aged, 80 and over , Humans , Male , Middle Aged , Case-Control Studies , Gene Frequency , Genetic Predisposition to Disease , Genotype , Interleukin-6 , Genetics , Matched-Pair Analysis , Pneumoconiosis , Genetics , Polymerase Chain Reaction , Polymorphism, Restriction Fragment LengthABSTRACT
<p><b>OBJECTIVE</b>To explore the relationship between the polymorphism of Interleukin-1 and the pneumoconiosis susceptibility.</p><p><b>METHODS</b>Eighty patients with silicosis and 45 with coal workers' pneumoconiosis (CWP) were selected while 125 male workers, Han nationality in the same workplace as the patients were selected as the controls. Between the patients and the control, the differences of age and cumulative length of service were less than five years and two years, respectively. The controls were exposed to dusts but did not suffer from pneumoconiosis. Moreover, the patients and the controls were paired by 1:1. DNA was extracted from leucocytes by the hydroxybenzene chloroform method. The polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) techniques and PCR were used to examine polymorphism of IL-1alpha (-889), IL-1beta (-511) and IL-1Ra (+2018) and variable number of tandem repeats (VNTR) of IL-1Ra. After the preliminary experiment, the most adaptive PCR reaction, the restriction enzyme digest and electrophoresis system were used.</p><p><b>RESULTS</b>The difference in IL-1alpha (-889) 1/2 + 2/2 between the pneumoconiosis patients and the controls was significant (P < 0.01). The result of conditional Logistic regression showed that heterozygote and allele 2 of IL-1a (-889) were risk factors of pneumoconiosis. The difference in the genotype frequencies of IL-1beta (-511) 1/2 + 2/2, IL-1Ra (+2018) 1/2 + 2/2 and IL-Ra VNTR1/2 + 2/2 between the patients and the controls were not statistically significant (P > 0.05).</p><p><b>CONCLUSION</b>IL-1alpha (-889) gene polymorphism is related to pneumoconiosis. Workers with IL-1alpha (-889) allele 2 are susceptible to the pneumoconiosis. The relationship between IL-1beta (-511), IL-1Ra (+2018), IL-1Ra VNTR genes polymorphisms and pneumoconiosis are not found.</p>