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OBJECTIVE: To compare the applicability of three risk assessment methods on occupational health risk assessment of chemical harmful factors in a resin anchorage production workshop. METHODS: A resin anchoring agent production workshop was selected as the research subject using the judgment sampling method. Contact ratio assessment method and comprehensive index method(both are semi-quantitative evaluation method) and qualitative risk assessment method were applied to estimate the occupational health risks of jobs involved with styrene, dibutyl phthalate and phthalic anhydride. The assessment was carried out, and the obtained risk level was standardized as the risk ratio. The evaluation results of these three methods were compared. RESULTS: The assessment results of exposure ratio method were 1-3, and those of comprehensive index method were 2-3. The risk ratio of the above methods after standardization is consistent with that before standardization. The result of qualitative risk assessment was 2-3, and the risk ratio after standardization was 3-4. The risk ratio after standardization was 1 level higher than that before standardization. Kappa analysis results showed that the result of contact ratio method and the comprehensive index method was in good agreement(Kappa=0.53, P<0.05). The qualitative risk assessment method was inconsistent with the contact ratio method and the comprehensive index method(Kappa values were 0.19 and 0.40, both P<0.05). CONCLUSION: Comprehensive index method is the preferred method for occupational health risk assessment of anchorage agent production workshop under the condition that detection results of occupational hazards can be obtained. Qualitative risk assessment method can be used when the test results cannot be obtained.
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Objective To study the inhibitory effect of T lymphocytes secreting EphrinAl-Caspase-3 in vivo and on the growth of cancer cells in nude mice with breast cancer. Methods Nude mice (n = 35) were inoculated with breast cancer cells to construct a nude mouse model of breast cancer. When the tumor volume reached 0. 1 cm3, 30 nude mice with average size tumor tissue were randomly divided into PBS group, uninfected adenovirus group, T lymphocyte infected with Ad-EphrinAl-Caspase-3 group, and intratumoral transplantation. Tumor size was measured every day 2 to 3. Three groups of tumor-bearing nude mice were selected. After the above-mentioned cell transplantation, the subcutaneous tumor tissue homogenate was obtained every day 2 to 3, and the content of EphrinAl-Caspase-3 was detected by ELISA. At the end of the experiment, the animals in each group were sacrificed by cervical dissection and sliced. The presence of T lymphocytes expressing green fluorescent protein was observed under a fluorescence microscope, and Caspase-3 and Ki-67 were detected by immunofluorescence. Results After one week of inoculation of breast cancer cells into nude mice, the presence of subcutaneous tumors could be touched by hand, which proved that the tumor-bearing animals of breast cancer cells were successfully modeled. On the 8th day after inoculation, the tumor volume of the nude mice in each group became larger, and the difference between the treatment group and the PBS group/T lymphocyte group was extremely significant ( P<0.05). Although the tumor volume of the T lymphocyte transplantation group was slower than that of the PBS control group, there was no statistically significant difference between the two. The expression of EphrinAl-Caspase-3 was detected in the EphrinAl-Caspase-3 treatment group on the 2nd day, reached the peak on the 8th day, and then the secretion decreased gradually. No expression of EphrinAl-Caspase-3 was detected in the PBS control group and the T lymphocyte group. The presence of dispersed green fluorescent protein-labeled EphrinAl-Caspase-3-T lymphocytes was observed in the tumor tissues of the treatment group, while the presence of green fluorescent protein was not detected in the PBS group and the T lymphocyte groups. In the infected cells of the treatment group, the proportion of Caspase-3 positive cells was up- regulated, and the proportion of Ki-67 positive cells was down-regulated. No expression of EphrinAl-Caspase-3 was detected in the PBS group and the T lymphocyte group. Conclusion EphrinAl-Caspase-3 can significantly inhibit the growth of breast cancer cells, thereby exerting an anti-tumor effect.
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The paper was aimed to explore the role of serum exosomes induced by hepatic ischemia/reperfusion (I/R) injury in the damage of hippocampus and cerebral cortex of rats. The male Sprague-Dawley (SD) rats were randomly divided into 4 groups: sham operation group (S), hepatic I/R injury group (I/R), serum exosomes from S group treatment group (ES) and serum exosomes from I/R group treatment group (EI). In ES group and EI group, 100 μL serum exosomes from S group and I/R group were injected into the normal rats through tail vein respectively. Another three normal rats were injected intravenously with serum exosomes labeled with PKH26 red fluorescence, and then the expression of fluorescence in the brain tissues was observed by immunofluorescence microscope. The morphology and size of exosomes were observed by transmission electron microscope, the expression of exosomes markers CD63 and CD9 was detected by Western blot, and the damage of liver and brain, levels of apoptosis and oxidative stress response in hippocampus and cerebral cortex were observed by serological and histological indexes. The results showed that the exosomes were a group of round or ovoid membranous vesicles, sized in 30-100 nm. Compared with that in S group, the content of serum exosomes in I/R group was increased (P < 0.05). Moreover, serum exosomes could go through the blood-brain barrier and enter the brain tissue freely through blood circulation. The index of liver function in I/R group was significantly higher than that in S group (P < 0.05). There was no significance in the degree of brain damage, apoptosis and oxidative stress in hippocampus and cerebral cortex between S group and ES group. Compared with those in S group and ES group, the serum levels of brain injury markers, apoptosis index (AI) and oxidative stress in hippocampus and cerebral cortex increased in I/R group and EI group (P < 0.05). Whereas, compared with those in I/R group, the above indicators in EI group decreased (P < 0.05). Therefore, hepatic I/R injury can lead to the damage of hippocampus and cerebral cortex, and the increased serum exosomes induced by hepatic I/R plays an important role.
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Animales , Masculino , Ratas , Isquemia Encefálica , Exosomas , Hipocampo , Hígado , Ratas Sprague-Dawley , Daño por ReperfusiónRESUMEN
OBJECTIVE@#To observe the application of modified traction therapy in traumatic atlantoaxial subluxation in adults.@*METHODS@#The clinical data of 31 patients with atlantoaxial subluxation treated from March 2018 to June 2019 were restropectively analyzed. There were 15 males and 16 females, aged from 18 to 68 years old with an average of 39 years old, including 10 cases of 18-40 years, 15 cases of 41-60 years, 6 cases of 51-68 years. The main manifestations of the patients were limited neck movement, pain, and atlantoaxial CT scan showed different degrees of atlantoaxial subluxation. Three dimensional multifunctional traction bed was used for traction for 2 min, relaxation for 10 s. The traction angle starts from the rearward extension of 5°-10° and weight from 3-6 kg. The weight increased by 1 kg every two days until the symptoms were improved. Traction time was 30 min twice a day and 10 days for a course of treatment. One course of treatment was performed in patients with 1-2 mm left and right equal width of atlantoaxial space, and two courses of treatment were performed in patients with 3-4 mm left and right equal width of atlantoaxial space, and the course of treatment could be increased to 3 months in especially patients with serious problems, such as 4 mm left and right equal width of atlantoaxial space and no improvement after conventional treatment. The criteria to evaluate the clinical effect was cure:no pain in the neck, normal range of neck movement, CT showed normal atlantoaxial space and odontoid process was in the middle, patients with normal neck movement were followed up 1 month after the end of treatment;improvement:neck pain was significantly improved and CT showed that the left and right atlantoaxial space was less than 1 mm in equal width.@*RESULTS@#Among the 31 patients, 17 cases were cured by one course of treatment, 11 cases were cured by 2 courses of treatment, and 2 caseswere improved.@*CONCLUSION@#The modified traction therapy has obvious effect on adult traumatic atlantoaxial subluxation, especially the subluxation of 3-4 mm equal width in left and right atlantoaxial space, and this method is safe and reliable with good efficacy and the patients without discomfort.
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Adolescente , Adulto , Anciano , Femenino , Humanos , Masculino , Persona de Mediana Edad , Adulto Joven , Articulación Atlantoaxoidea , Luxaciones Articulares , Apófisis Odontoides , Fusión Vertebral , TracciónRESUMEN
In this experiment,the antioxidant capacity of raspberry extract and the protective effect on liver injury induced by ConA in mice were investigated. Balb/C male mice were randomly divided into six groups: normal group,model group,bicyclol control group( 200 mg·kg~(-1)),low-dose raspberry extract group( 200 mg·kg~(-1)),middle-dose raspberry extract group( 400 mg·kg~(-1)),and highdose raspberry extract group( 800 mg·kg~(-1)). Each group was intragastrically administered with drugs according to the body weight once a day. Seven days later,all of the groups except for the normal group were treated with ConA( 20 mg·kg~(-1)) through tail vein injection to establish the acute liver injury model. The mice were put to death 8 hours later. The organ indexes were calculated. These rum levels of ALT,AST and LDH and the activities of SOD,CAT,GSH and MDA in liver tissue were detected. HE staining was used to observe the pathological changes of liver tissue in mice. Western blot was used to detect the expressions of Bax,Bcl-2,Nrf2 and Keap-1. The antioxidant capacity of raspberry extract was measured by CAA assay. The results showed that,raspberry extract had a strong antioxidant capacity. Simultaneously,compared with the model group,raspberry extract can significantly improve the pathological conditions of liver,and significantly reduce ALT,AST and LDH activities in serum of liver injury mice( P<0. 01). The activities of SOD,CAT in liver homogenate supernatant were significantly increased in the high-dose group,the content of GSH increased,while the content of MDA was sharply declined in the high-dose group( P<0. 01). Meanwhile,raspberry extract down-regulated the expressions of Bax and Keap-1 and up-regulated the expressions of Bcl-2 and Nrf2. CAA showed that the compound raspberry extract had a strong antioxidant capacity. Therefore,raspberry extract has an obvious protective effect on acute liver injury induced by ConA in mice.
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Animales , Masculino , Ratones , Antioxidantes , Enfermedad Hepática Inducida por Sustancias y Drogas , Hígado , Ratones Endogámicos BALB C , Sustancias Protectoras , RubusRESUMEN
Peroxisome proliferator-activated receptor-γ coactivator 1α (PGC-1α) is a transcriptional co-activator involved in mitochondrial biogenesis, respiratory capacity, and oxidative phosphorylation (OXPHOS). PGC-1α plays an important role in cellular metabolism and is associated with tumorigenesis, suggesting an involvement in cell cycle progression. However, the underlying mechanisms mediating its involvement in these processes remain unclear. To elucidate the signaling pathways involved in PGC-1α function, we established a cell line, CH1 PGC-1α, which stably overexpresses PGC-1α. Using this cell line, we found that over-expression of PGC-1α stimulated extra adenosine triphosphate (ATP) and reduced reactive oxygen species (ROS) production. These effects were accompanied by up-regulation of the cell cycle checkpoint regulators CyclinD1 and CyclinB1. We hypothesized that ATP and ROS function as cellular signals to regulate cyclins and control cell cycle progression. Indeed, we found that reduction of ATP levels down-regulated CyclinD1 but not CyclinB1, whereas elevation of ROS levels down-regulated CyclinB1 but not CyclinD1. Furthermore, both low ATP levels and elevated ROS levels inhibited cell growth, but PGC-1α was maintained at a constant level. Together, these results demonstrate that PGC-1α regulates cell cycle progression through modulation of CyclinD1 and CyclinB1 by ATP and ROS. These findings suggest that PGC-1α potentially coordinates energy metabolism together with the cell cycle.
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Adenosina Trifosfato/metabolismo , Ciclo Celular/fisiología , Ciclina B1/metabolismo , Ciclina D1/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Factores de Transcripción/metabolismo , Regulación del Desarrollo de la Expresión Génica/fisiología , Células HEK293 , Humanos , Coactivador 1-alfa del Receptor Activado por Proliferadores de Peroxisomas gammaRESUMEN
<p><b>BACKGROUND</b>Urination disorders are common in Parkinson's disease (PD) and respond poorly to medication. This study aimed to analyze the risk factors for urination disorders in PD.</p><p><b>METHODS</b>Ninety-one patients with PD (aged 34-83 years old) were recruited. Patients were assessed with the Unified PD Rating Scale (UPDRS), Hoehn and Yahr stage, Pittsburgh Sleep Quality Index (PSQI), Hamilton Depression Rating Scale (HAMD), and Hamilton Anxiety Scale (HAMA). Micturition number was recorded, and Type B ultrasound was used to evaluate residual urine. Statistics was performed using binary logistic regression, bivariate correlations, and Chi-square and t-tests.</p><p><b>RESULTS</b>Of 91 patients, urinary dysfunction occurred in 55.0%. Among these, 49.5% suffered with nocturia, 47.3% with pollakiuria. Nocturia number had a positive linear relationship with HAMA score (odds ratio [OR] = 0.340, P = 0.001), HAMD score (OR = 0.323, P = 0.002), duration of L-dopa medication (OR = 0.328, P = 0.001), dose of L-dopa (OR = 0.273, P = 0.009), UPDRS-II (OR = 0.402, P = 0.000), UPDRS-III score (OR = 0.291, P = 0.005), and PSQI score (OR = 0.249, P = 0.017). Micturition number over 24 h was positively associated with HAMA (OR = 0.303, P = 0.004) and UPDRS-II scores (OR = 0.306, P = 0.003). Of patients with residual urine, 79.3% had a volume of residual urine <50 ml. Residual urine was present in 44.4% of the patients with nocturia, 46.5% of the patients with pollakiuria, and 80.0% of the patients with dysuria. More men than women had residual urine (35.2% male vs. 13.3% female; P = 0.002).</p><p><b>CONCLUSIONS</b>Nocturia and pollakiuria were common micturition symptoms in our participants with PD. Nocturia was associated with depression, anxiety, sleep problems, and severity of PD. Pollakiuria was associated with anxiety and severity of PD. Male patients were more prone to residual urine and pollakiuria.</p>
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Adulto , Anciano , Anciano de 80 o más Años , Femenino , Humanos , Masculino , Persona de Mediana Edad , Nocturia , Enfermedad de Parkinson , Factores de Riesgo , Trastornos UrinariosRESUMEN
<p><b>OBJECTIVE</b>To observe the effects of low-intensity pulsed ultrasound (LIPUS) on repairing extracellular matrix in rabbit knee osteoarthritis and analyze its mechanism.</p><p><b>METHODS</b>Sixty adult female rabbits with an average weight of (2.0 ± 0.2) kg, were divided randomly into two groups (experimental group and control group, 30 rabbits in each group). All rabbits were replicated in right knees by Hulth method for knee osteoarthritis model. Two weeks after operation, the rabbits in experimental group were treated with LIPUS, and the ultrasonic frequency was (800 ± 5%)KHz and the maximum intensities of spatially averaged and time averaged (SATA) was (50 ± 10%) mw/cm2, for 1 time a day and every time 20 min, while the rabbits in control group were treated with sham LIPUS,the same operation with experimental group but without energy output. At the 2, 4, 8 weeks after treatment, 10 rabbits in each group were randomly killed for each time. The general changes of cartilage and its histopathological changes by HE staining were observed; the expression of collagen type II, proteoglycan, MMP-3, 7, 13 in cartilage were analyzed by immunohistochemical and RT-PCR technique; and the expression of NO in cartilage was analyzed by nitrate reduction method.</p><p><b>RESULTS</b>On the same observed time point, the damage degree of cartilage in experimental group was slighter than that of control group (P < 0.01), the expression of MMP-3, 7, 13 and NO in cartilage in experimental group was lower than that of control group (P < 0.01) while collagen type II and proteoglycan was higher than that of control group (P < 0.01).</p><p><b>CONCLUSION</b>Low-intensity pulsed ultrasound can repair the damaged cartilage by reducing the expression of MMP-3, 7, 13, inhibiting the secretion of NO and promoting the synthesis of collagen type II and proteoglycan in cartilage.</p>
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Animales , Femenino , Conejos , Cartílago Articular , Patología , Colágeno Tipo II , Matriz Extracelular , Metabolismo , Metaloproteinasas de la Matriz , Óxido Nítrico , Osteoartritis de la Rodilla , Metabolismo , Terapéutica , Terapia por Ultrasonido , MétodosRESUMEN
<p><b>OBJECTIVE</b>To screening mutations of exons 15, 18 and 26 of sodium channel Nav1.7 (SCN9A) gene, and to assess its association with pain related to Parkinsonism.</p><p><b>METHODS</b>Respectively, 101 patients with primary Parkinson's disease (PD) and 104 similar-aged volunteers without PD were recruited from March, 2008 to January, 2011. Mutations of above 3 exons in SCN9A gene was detected with PCR and direct sequencing. For 100 patients with Parkinsonism, the pain was scored with a McGill pain rating scale. Statistical analysis was performed with SPSS.</p><p><b>RESULTS</b>The prevalence of pain in Parkinsonian was 57%. 43.86% patients with pain were males, and 56.14% were females. Based on Chaudhuri criteria, the pain symptoms may be classified as musculoskeletal pain (10.52%), radicular pain (10.52%), dyskinesis pain (54.38%), pain from akathisia and restlessness (14.04%), dyskinesis combined with radicular pain (5.26%), skeletal muscles pain and headache (1.75%), and arthralgia (3.50%). Two missense mutations were identified, which included 2794A/C (0.941/0.059) (rs12478318) (M932L) in exon 15 and 3448C/T (0.988/0.012) (rs6746030) (R1150W) in exon 18. The wild type A/C for the 2794 locus had a higher prevalence in PD patients with pain, but this was not statistically different. All of the 5 heterozygotes for 3448 (C/T) were found in Parkinsonian patients with pain. No homozygotes were found.</p><p><b>CONCLUSION</b>The prevalence of pain was higher in Parkinsonian patients than general population, and the proportion of males to females was similar. More patients have suffered dyskinesis pain. A 3448 (C/T) mutation of SCN9A gene may be related to pathogenesis of pain in Parkinsonism.</p>
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Adulto , Anciano , Anciano de 80 o más Años , Femenino , Humanos , Masculino , Persona de Mediana Edad , Alelos , Secuencia de Bases , Exones , Frecuencia de los Genes , Genotipo , Mutación , Genética , Dolor , Epidemiología , Enfermedad de Parkinson , Genética , PrevalenciaRESUMEN
OBJECTIVE: To prepare uniform-sized rifabutin-loaded PLGA microspheres and evaluate the characteristics of the microspheres. METHODS: The particle size uniformity of the microspheres prepared by premix membrane emulsification method and homogenization method was compared. Preparation technology was optimized by orthogonal design. Then the drug loading, entrapment efficiency and in vitro drug release were studied. RESULTS: The average particle size and PDI of the microspheres prepared by homogenization method were (4242±175.6) nm and 0.330. The microspheres prepared by premix membrane emulsification method were round, complete and smooth, and their particle sizes were well-distributed with mean value of (1330±64) nm and PDI of 0.168. Besides, the drug loading was (46.83±0.29)% and the entrapment efficiency was (93.66±0.58)%. Accumulative drug release rate was 66% in 15 d. CONCLUSION: The rifabutin-loaded PLGA microspheres prepared by premix membrane emulsification method have narrow particle size distribution, high drug loading, high entrapment efficiency, and ideal in vitro sustained-release effect.
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<p><b>OBJECTIVE</b>To investigate the effects of hypoxia during the prenatal period and its later repercussions on sexual behavior and the sex hormone secretion of male rats.</p><p><b>METHODS</b>Experimental animals were divided into three groups randomly: control group, which was kept at normal atmospheric pressure, and two stress groups exposed to a simulated altitude equivalent to 3000 m and 5000 m, respectively. Stress groups were exposed to hypoxic circumstance at their final week of gestation in animal decompression chamber.</p><p><b>RESULTS</b>Adulthood, males exposed to hypoxic stress during the prenatal period were able to mate with normal females, but these treated males exhibited decreased male sexual behavior. Decreased anogenital distances were observed in male offspring, and presented reductions of plasma testosterone levels, increase of plasma corticosterone levels, but no notable alteration in the organ index.</p><p><b>CONCLUSION</b>These results indicate that exposure to hypoxia in the later stages of pregnancy may have a long-term effect on the fertility and sexual behavior of male offspring.</p>
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Animales , Femenino , Masculino , Embarazo , Ratas , Hipoxia , Efectos Tardíos de la Exposición Prenatal , Ratas Sprague-Dawley , Conducta Sexual Animal , Fisiología , Estrés Fisiológico , Fisiología , Testosterona , SangreRESUMEN
<p><b>OBJECTIVE</b>To ascertain the optimal harvest time of Erigeron breviscapus.</p><p><b>METHOD</b>The dry matter weight accumulation of different organs in growth process and contents of scutellarin and coffeic acid ester in whole plant of E. breviscapus were determined.</p><p><b>RESULT</b>The number of leaves per plant, the dried weight of single leaf and dry matter weight of whole plant and different organs reached the highest after seedling 130-140 d. The content of scutellarin gradually decreased with growth period, and sharply decreased after seedling 140 d. The content of coffeic acid ester varied irregularly with growth period.</p><p><b>CONCLUSION</b>The optimal harvest time of E. breviscapus is in early bloom period after seedling 130 d.</p>
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Apigenina , Biomasa , Erigeron , Química , Jardinería , GlucuronatosRESUMEN
<p><b>OBJECTIVE</b>To search for a clinically effective therapy for periarthritis of shoulder.</p><p><b>METHODS</b>One hundred and forty cases were randomly divided into a treatment group and a control group, 70 cases in each group. The treatment group were treated with "Jianying point" -injection combined with massage, and the control group with routine acupuncture therapy. The pain scores and the comprehensive scores of motor function before and after treatment in the two groups were compared between the two groups.</p><p><b>RESULTS</b>The different value of the pain scores before and after treatment was 31.54+/-2.82 in the treatment group and 15.03+/-4.07 in the control group, with a very significant difference between the two groups (P<0.01); after treatment, the comprehensive score of motor function of the shoulder joint was 139.64+/-12.34 in the treatment group and 80.32+/-16.29 in the control group, with a very significant difference between the two groups (P<0.01).</p><p><b>CONCLUSION</b>"Jiunying point" -injection combined with massage has clinically an obvious effect on periarthritis of shoulder.</p>
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Adulto , Anciano , Femenino , Humanos , Masculino , Persona de Mediana Edad , Puntos de Acupuntura , Dexametasona , Inyecciones , Masaje , Periartritis , Terapéutica , Articulación del Hombro , Dolor de Hombro , Terapéutica , Vitamina B 12RESUMEN
The dorsal horn of the spinal cord consists of distinct laminae that serve as a pivotal region for relaying a variety of somatosensory signals such as temperature, pain, and touch. The molecular mechanisms underlying the development of the dorsal horn are poorly understood. To define a molecular map of the dorsal horn circuit, we have profiled dorsal horn-enriched (DHE) gene expression in dorsal spinal cords on embryonic day 15.5 (E15.5) by genome-wide microarray and smart subtractive screening based on polymerase chain reaction (PCR). High-throughput in situ hybridization (ISH) was carried out to validate the expression of 379 genes in the developing dorsal spinal cord. A total of 113 DHE genes were identified, of which 59% show lamina-specific expression patterns. Most lamina-specific genes were expressed across at least two laminae, however. About 32% of all DHE genes are transcription factors, which represent the largest percentage of the group of all DHE functional classifications. Importantly, several individual lamina-specific transcription factors such c-Maf, Rora, and Satb1 are identified for the first time. Epistasis studies revealed several putative effectors of known DHE transcription factors such as Drg11, Tlx3(Rnx), and Lmx1b. These effector genes, including Grp, Trpc3, Pcp4, and Enc1, have been implicated in synaptic transmission, calcium homeostasis, and structural function and thus may have similar roles in the dorsal horn. The identification of a large number of DHE genes, especially those that are lamina specific, lays a foundation for future studies on the molecular machinery that controls the development of the dorsal horn and on functional differences of these distinct laminae in the dorsal spinal cord.
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Células del Asta Anterior/metabolismo , Regulación del Desarrollo de la Expresión Génica , Hibridación Genética , Análisis de Secuencia por Matrices de Oligonucleótidos/métodos , Células del Asta Posterior/metabolismo , Médula Espinal/embriología , Animales , ADN Complementario/genética , Inmunohistoquímica , Hibridación in Situ , Ratones , Ratones Endogámicos C57BL , Modelos Biológicos , Sondas ARN , Médula Espinal/citología , Médula Espinal/metabolismo , Factores de Transcripción/genética , Factores de Transcripción/metabolismoRESUMEN
OBJECTIVE To investigate existed or potential dangerous incidents in the daily work of sterilization and supply center, and establish countermeasures against them. METHODS The causes of dangerous incidents confirmed by nursing department from Jun 2006 to Oct 2007 were analyzed. RESULTS The most common dangerous incidents were nonstandard clinical self-prepared operation packs, non-fully cleared appliance and the non-turnoff of storage aperture. And the rate of the incidents was 18.6%, 10.9% and 9.2%, respectively. Direct dangerous incidents accounted for 72.3% and indirect dangerous incidents accounted for 27.7% in the total number of dangerous incidents. CONCLUSIONS The dangerous incidents can be reduced effectively by adopting relevant measures, enhancing risk surveillences and supervising the high-risk parts.
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<p><b>OBJECTIVE</b>To screen for polymorphisms in alpha 4 subunit (principal subunit of nAChR) gene (CHRNA4).</p><p><b>METHODS</b>DNA was extracted from leukocytes of all subjects including 100 healthy senior people and 100 patients with Parkinson's disease (PD). The exons and adjacent intron regions of CHRNA4 were amplified with PCR. SNPs were screened by denatured high performance liquid chromatography (DHPLC) techniques and restriction fragment length polymorphisms. Potential mutations were confirmed by sequencing.</p><p><b>RESULTS</b>Total 10 polymorphisms were detected and identified in coding and adjacent intron regions of nAChR alpha 4 gene, that are 420C/T (0.873/0.127), 870C/T (0.828/0.172), 1440A/C (0.858/0.142), 1860C/T (0.738/0.262), 1890C/T (0.605/0.395), intron 5 +14T/C (0.553/0.447), intron 2 +22G/A (0.873/0.127), intron 3 +182 Del22bp (0.813/0.187), 1758C/T and 1809C/T (reference for coding sequence is GenBank SNPs 000744), of which the last three are novel mutations. PD patients appeared higher frequency of deletion in intron 3+182(0.235) than normal controls (0.140)(P=0.015).</p><p><b>CONCLUSION</b>nAChR alpha 4 gene is polymorphic. PD patients take higher frequency of intron3+182 Del 22 bp.</p>
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Anciano , Femenino , Humanos , Masculino , Pueblo Asiatico , Genética , Secuencia de Bases , Datos de Secuencia Molecular , Enfermedad de Parkinson , Genética , Polimorfismo Genético , Receptores Nicotínicos , Genética , Análisis de Secuencia de ADNRESUMEN
<p><b>BACKGROUND</b>The stage-specific expression of genes is one of the most characteristics of parasites. It has been found that a lot of genes of Spriometra erinaceieuropaei are specifically expressed in pleroceroid in large amount, but not expressed when the plerocercoid development into adult worm. The study is to screen other stage-specific ecpression genes of plerocercoid of Spirtmetra erinceieuropaei.</p><p><b>METHODS</b>RNA was separately extracted by acid guanidinium thiocyanate-phenol-chloroform from plerocercoids and adult worms of Spirometra erinaceieuropaei, DNA contaminated in the RNA was digested by RNase-free DNase. After the RNA was reverse transcripted to cDNA using T12MA, T12MC, T12MG and T12MT anchor-primers, PCR was done using the same T12MN and one random primer with alpha 35S-dATP in the system. The PCR products were fractionated on an 8% denatured polyacrylamide gel. Differential bands of the plerocercoid found in the gel were cut out, amplified by PCR and sequenced. Northern hybridization was used to identify the stage-specific expression genes.</p><p><b>RESULTS</b>Eleven differential bands were selected from the gel and classified into 3 kinds of gene fragments by hybridization, after they were amplified by PCR. Fragments 1 (238 bp) and 2 (383 bp), were confirmed by Northern hybridization, as being expressed in the plerocercoid. However, fragment 3 (433 bp), was expressed in both the plerocercoid and the adult worm. Data from the 3 gene fragments underwent homological analysis in GenBank. The sequence which was homologous with fragments 1 and 2 was not found, but fragment 3 had high homology with many kinds of 28S rRNA.</p><p><b>CONCLUSIONS</b>The gene expressions of plerocercoids are different from adult worms because they live in different hosts. Two types of different gene fragments from the plerocercoid were found by mRNA differential display technique.</p>