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The potential for off-target mutations is a critical concern for the therapeutic application of CRISPR-Cas9 gene editing. Current detection methodologies, such as GUIDE-seq, exhibit limitations in oligonucleotide integration efficiency and sensitivity, which could hinder their utility in clinical settings. To address these issues, we introduce OliTag-seq, an in-cellulo assay specifically engineered to enhance the detection of off-target events. OliTag-seq employs a stable oligonucleotide for precise break tagging and an innovative triple-priming amplification strategy, significantly improving the scope and accuracy of off-target site identification. This method surpasses traditional assays by providing comprehensive coverage across various sgRNAs and genomic targets. Our research particularly highlights the superior sensitivity of induced pluripotent stem cells (iPSCs) in detecting off-target mutations, advocating for using patient-derived iPSCs for refined off-target analysis in therapeutic gene editing. Furthermore, we provide evidence that prolonged Cas9 expression and transient HDAC inhibitor treatments enhance the assay's ability to uncover off-target events. OliTag-seq merges the high sensitivity typical of in vitro assays with the practical application of cellular contexts. This approach significantly improves the safety and efficacy profiles of CRISPR-Cas9 interventions in research and clinical environments, positioning it as an essential tool for the precise assessment and refinement of genome editing applications.
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Sistemas CRISPR-Cas , Edición Génica , Células Madre Pluripotentes Inducidas , Humanos , Edición Génica/métodos , Células Madre Pluripotentes Inducidas/metabolismo , Células Madre Pluripotentes Inducidas/efectos de los fármacos , Células Madre Pluripotentes Inducidas/citología , Mutación , ARN Guía de Sistemas CRISPR-Cas/genética , Células HEK293RESUMEN
OBJECTIVE@#To assess the efficacy on relieving pain and improving the range of motion of shoulder joint in post-stroke shoulder-hand syndrome of phlegm-stasis obstruction in treatment of the combined therapy of eye acupuncture, Tengliao (Chinese herbal warm dressing technique) and rehabilitation training (eye acupuncture + Tengliao + rehabilitation) as compared with the combined treatment of Tengliao and rehabilitation training (Tengliao + rehabilitation) and the simple rehabilitation training (rehabilitation).@*METHODS@#A total of 356 patients with post-stroke shoulder-hand syndrome of phlegm-stasis obstruction were randomized into an eye acupuncture + Tengliao + rehabilitation group (group A, 122 cases, 2 cases dropped off), a Tengliao + rehabilitation group (group B, 120 cases, 3 cases dropped off) and a rehabilitation group (group C, 114 cases, 1 case dropped off). In the group C, the basic treatment was combined with routine rehabilitation training. In the group B, on the base of the treatment as the group C, Tengliao was exerted. A medical bag composed of over 20 Chinese herbal materials was heated and dressed at the affected area, 30 min each time, 5 times weekly. In the group A, besides the treatment as the group B, eye acupuncture was applied to heart region, kidney region, upper jiao region and lower jiao region, 30 min each time, 5 times weekly. The treatment lasted 28 days in all of three groups. Separately, before treatment, in 7, 14, 21 and 28 days of treatment, as well as in 14 days after treatment of follow-up, the score of visual analogue scale (VAS) for pain, the score of guides to evaluation of permanent impairment (GEPI) and the score of National Institutes of Health stroke scale (NIHSS) were observed in each group.@*RESULTS@#The scores of VAS, GEPI and NIHSS were all improved with the treatment lasting in the three groups (P<0.000 1). In 7, 14, 21 and 28 days of treatment and in follow-up as well, VAS scores in the group A were all lower than the group C (P<0.05). After 14 days of treatment, GEPI score showed increasing trend, while NIHSS score showed decreasing trend in the group A compared with the group B. Before treatment, GEPI score was lower and NIHSS score was higher in the group A compared with the group C (P<0.05). It was suggested that the illness was slightly serious in the group A. After propensity score matching, in 14, 21 and 28 days as well as in follow-up, GEPI scores in the group A were higher than the group C respectively (P<0.05). Regarding NIHSS score at each time point, the difference had no statistical significance between the group A and the group C (P>0.05).@*CONCLUSION@#The combined therapy of eye acupuncture, Tengliao and rehabilitation training obtains a better efficacy on post-stroke shoulder-hand syndrome of phlegm-stasis obstruction as compared with rehabilitation training.
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Humanos , Puntos de Acupuntura , Terapia por Acupuntura/métodos , Dolor , Distrofia Simpática Refleja/terapia , Accidente Cerebrovascular/complicaciones , Rehabilitación de Accidente Cerebrovascular , Resultado del TratamientoRESUMEN
Aim To investigate the effects of Evodiamine (EVO) on proliferation and apoptosis of human leukemia cell line K562 and its potential mechanisms. Methods K562 cells were treated with EVO at different concentrations (0, 1, 2, 4, 8, 16, 32, 64 jxmol • L
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BackgroundWe aim to investigate the profile of acute antibody response in COVID-19 patients, and provide proposals for the usage of antibody test in clinical practice. MethodsA multi-center cross-section study (285 patients) and a single-center follow-up study (63 patients) were performed to investigate the feature of acute antibody response to SARS-CoV-2. A cohort of 52 COVID-19 suspects and 64 close contacts were enrolled to evaluate the potentiality of the antibody test. ResultsThe positive rate for IgG reached 100% around 20 days after symptoms onset. The median day of seroconversion for both lgG and IgM was 13 days after symptoms onset. Seroconversion of IgM occurred at the same time, or earlier, or later than that of IgG. IgG levels in 100% patients (19/19) entered a platform within 6 days after seroconversion. The criteria of IgG seroconversion and > 4-fold increase in the IgG titers in sequential samples together diagnosed 82.9% (34/41) of the patients. Antibody test aided to confirm 4 patients with COVID-19 from 52 suspects who failed to be confirmed by RT-PCR and 7 patients from 148 close contacts with negative RT-PCR. ConclusionIgM and IgG should be detected simultaneously at the early phase of infection. The serological diagnosis criterion of seroconversion or the >; 4-fold increase in the IgG titer is suitable for a majority of COVID-19 patients. Serologic test is helpful for the diagnosis of SARS-CoV-2 infection in suspects and close contacts.
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OBJECTIVE: To investigate the clinical features and risk factors of multidrug-resistant bloodstream infection in children with acute leukemia. METHODS: The clinical data of 121 blood culture-positive patients with acute leukemia admitted from January 1,2013 to September 30,2018 to Department of Pediatrics,Zhujiang Hospital of Southern Medical University were analyzed retrospectively. RESULTS: Of the 121 patients with acute leukemia infected with bacterial bloodstream,55 were in the multidrug-resistant(MDR)group and 66 in the non-multidrug-resistant(non-MDR)group. There were 31 grampositive bacteria in the MDR group. The top three strains were coagulase-negative Staphylococci,Staphylococcus aureus and Streptococcus mutans. Escherichia coli was the main strain of gram-negative bacteria. Logistic analysis suggested that MDR bloodstream infection was more likely to occur in the patiens with AML(P=0.038;OR 2.505;95%CI 1.036—6.058)and at induction chemotherapy stage(P=0.038;OR 2.226;95%CI 1.045—4.774). Other high-risk factors included neutropenic dysplasia >7 d before fever(P=0.003;OR 3.36;95%CI 1.520—7.428),hemoglobin 7 days,hemoglobin<70 g/L and platelet<20×109/L are risk factors for MDR bloodstream infection. The inflammation response is severe MDR bloodstream infections,which may result in longer anti-infective treatments and a worse prognosis.
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Through deeply analyzing the classification and coding of medical devices system,as well as existing problems in China,it proposed to build a medical devices classification and coding System,which was unified from state level and a platform for collection as well as surveillance for the information of medical devices,so as to improve the standardized management of the medical devices in China.
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To study the effect of Shenqi Dihuang decoction on inflammatory factor, renal function and microcirculation in patients with early diabetic nephropathy. A total of 205 cases of patient with early diabetic nephropathy treated in our hospital from March 2014 to April 2016 were selected and divided into two groups according to the admitted order, with 103 cases in clinical group and 102 in control group. Patients in control group were treated with melbine and captopril, which may be adjusted according to the clinical symptom. The clinical group was given Shenqi Dihuang decoction. Then the clinical efficady, inflammatory factors, renal function, endothelial function and hemorheology index were compared. Compared with 77.45% in the control group, the total effective rate of the clinical group was 92.23%. There was a significant increase (<0.05). The comparison of the values of IL-6, IL-8, TNF-α and CRP between the two groups before and after treatment showed significant differences. The values of inflammatory factors in treatment group were lower than in control group (<0.05). The comparison of the values of β2-MG, Cys-C and urine m-ALB between the two groups before and after treatment showed significant differences. The values of renal function in treatment group were lower than those in control group (<0.05). Compared with before treatment, ET-1 of the two groups after treatment decreased, while NO increased (<0.05). Compared with the control group, the value of ET-1 in patients of the experimental group was lower after treatment, while NO was higher (<0.05). The comparison of the values of whole blood viscosity, plasma viscosity, whole blood reduction viscosity, platelet aggregation rate and fibrinogen between the two groups before and after treatment showed significant differences. The value of hemorheology index in treatment group was lower than that in control group (<0.05). Shenqi Dihuang decoction has a better effect on patients with early diabetic nephropathy. It can significantly intervene with inflammatory response, reduce proteinuria, protect the renal function of patients, and improve the patient's vascular endothelial function and blood rheology, so as to make microcirculation to recover to the normal level.
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<p><b>OBJECTIVE</b>To study the efficacy of modified Wuzhuyu Decoction Granule (, MWDG) in the treatment of migraine patients with cold and stasis obstructing meridian syndrome.</p><p><b>METHODS</b>This study was a randomized, double-blind, placebo-controlled trial. A total of 78 migraine patients with cold and stasis obstructing meridian syndrome were recruited and randomly assigned by a ratio of 2:1 into a treatment group (51 cases) and a placebo group (27 cases). Patients in the treatment group were treated with MWDG while placebo granules were applied in the control group. The treatment course lasted for 12 weeks with a follow-up of 4 weeks. The primary outcome measures included frequency and days of migraine attacks and the secondary outcome measures were analgesics consumption and visual analogue scale (VAS) scores. All outcome assessments were conducted respectively at baseline, the 4th, 8th and 12th week, and the end of follow-up.</p><p><b>RESULTS</b>In the treatment group, significant decrease in frequency of migraine attacks were observed since the 4th week and that of analgesics consumption since the 8th week (both P<0.05). While, in the placebo group, significant decrease in frequency of migraine attacks were observed since the 8th week and that of analgesics consumption since the 12th week (both P<0.05). No significant decrease in days of migraine attacks and VAS scores of migraine pain were observed in both groups. Between the two groups, there were significant differences in VAS scores and intensity of pain appeared in the 8th week (P<0.05). However, no significant differences were found in days and frequency of migraine attacks and analgesics consumption (P>0.05).</p><p><b>CONCLUSIONS</b>MWDG was probably effective in the treatment of migraine especially for alleviating pain intensity. Furthermore, MWDG could reduce the frequency of migraine attacks and analgesics consumption sooner than the placebo.</p>
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Adulto , Femenino , Humanos , Masculino , Analgésicos , Usos Terapéuticos , Demografía , Método Doble Ciego , Medicamentos Herbarios Chinos , Usos Terapéuticos , Meridianos , Trastornos Migrañosos , Quimioterapia , Dimensión del Dolor , Pacientes Desistentes del Tratamiento , Placebos , Síndrome , Resultado del TratamientoRESUMEN
The paper summarizes the relevant documents of the new payment fee schedule released in China,and elaborates the status of the management of the new payment fee schedule management,coupled with analysis of the new payment fee schedule issued by the provinces (autonomous regions and municipalities),which helps to put forward policy-oriented suggestions for the management of the new payment fee schedule.
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<p><b>OBJECTIVE</b>To investigate the value of hemoglobin A(HbA) for screening thalassemia.</p><p><b>METHODS</b>A total of 2 000 adults' peripheral blood samples from Guangdong Women and Children Hospital from June 2013 to January 2014 were collected. The hemoglobin A(HbA) level was analyzed by the full automatic capillary electrophoresis technique, and the genotypes of thalassemia were detected.</p><p><b>RESULTS</b>The optimal cutoff values of HbAfor screening silent α-thalassemia, α-thalassemia trait, intermedia α-thalassemia and β-thalassemia trait were 2.85%, 2.65%, 2.25% and 3.45%, respectively; the areas under receiver operator characteristic (ROC) curve were 0.709, 0.839, 0.979 and 0.997 respectively; the sensitivities were 0.481, 0.721, 0.953 and 0.994, and the specificities were 0.846, 0.837, 0.929 and 0.969 respectively.</p><p><b>CONCLUSION</b>The optimal cutoff values of HbAfor screening different type of thalassemia based on our laboratory data are established by using ROC curve. According to the area under ROC curve, a satisfactory accuracy for screening intermedia α-thalassemia and β-thalassemia trait can be achieved by detecting hemoglobin Alevel.</p>
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<p><b>OBJECTIVE</b>To observe the inhibitory effect of different extract fractions from Longdan Xiegan decoction on biofilms of Candida albicans, and discuss its possible mechanism.</p><p><b>METHOD</b>The micro-dilution method and the XTT reduction assay were adopted to explore the antifungal activity of different extract fractions from Longdan Xiegan decoction and detect the inhibitory effect of different extracts on biofilms of C. albicans. The expression quantity of the adhesion related gene ALS1 and hypha formation SUN41 were detected by qRT-PCR.</p><p><b>RESULT</b>The MICs of extracts from Longdan Xiegan decoction, including petroleum ether, water, butanol, methanol and ethyl acetate, against C. albicans were > 1 000, > 1 000, > 1 000, 125, 125 mg x L(-1). The SMIC50 against biofilms of C. albicanswere > 1 000, > 1000, > 1 000, 500, 500 mg x L(-1). The SMIC50 were > 1 000, > 1 000, > 1 000, > 1 000 and 1 000 mg x L(-1). 1 000 mg x L(-1) ethyl acetate extracts could considerably inhibit the expression of the adhesion related gene ALS1 and hypha formation SUN41.</p><p><b>CONCLUSION</b>The ethyl acetate extract showed the greatest activity against Candida albicans biofilms.</p>
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Humanos , Antifúngicos , Farmacología , Biopelículas , Candida albicans , Candidiasis , Quimioterapia , Microbiología , Medicamentos Herbarios Chinos , Farmacología , Hifa , Pruebas de Sensibilidad MicrobianaRESUMEN
<p><b>OBJECTIVE</b>To study the protective impact of tea polyphenols (TP) on the injury of fibrinolytic functions induced by high-methionine dietary in rats.</p><p><b>METHODS</b>50 male Wistar rats were divided by stratified based on body weight into 5 groups with 10 in each group: namely control group, model group, low-dose TP group, medium-dose TP group and high-dose TP group. The rats in model group and TP groups were fed with 3% methionine dietary, control group rats with routine diet. In addition, rats in low-dose, medium-dose and high-dose TP groups were treated with TP at 50, 100 and 200 mg/kg dosage respectively by gavages every day, control group and model group rats were given with same amount distilled water. The animals were sacrificed after 8 weeks. The levels of tissue-type plasminogen activator (t-PA) and type-1 plasminogen activator inhibitor (PAI-1) in plasma were determined by ELISA assays, mRNA levels of t-PA and PAI-1 in aortic arch were detected by RT-PCR, t-PA and PAI-1 expression in aortic arch were detected by immunohistochemistry strept-avidin-biotin complex (SABC).</p><p><b>RESULTS</b>After experiment, the t-PA expression of aortic arch in control group, model group, low-dose TP group, medium-dose TP group and high-dose TP group were 133.03 ± 10.14, 95.46 ± 11.08, 111.97 ± 11.91, 130.23 ± 10.80, 139.39 ± 9.41 (F = 14.15, P < 0.01), respectively, and the PAI-1 expression were 90.91 ± 8.67, 166.76 ± 12.18, 139.63 ± 12.71, 134.66 ± 13.19, 109.49 ± 10.82 (F = 31.44, P < 0.01). The t-PA concentration of plasma were (10.69 ± 1.26), (6.13 ± 0.92), (8.56 ± 1.19), (9.69 ± 0.92), (11.97 ± 1.08) ng/ml, respectively (F = 41.98, P < 0.01), and the PAI-1 concentration of plasma were (6.31 ± 0.81), (16.98 ± 1.27), (11.39 ± 0.82), (8.46 ± 0.67), (8.08 ± 0.91) ng/ml, respectively (F = 207.74, P < 0.01). The mRNA levels of t-PA in aortic arch were 1.12 ± 0.02, 0.75 ± 0.14, 1.01 ± 0.09, 0.95 ± 0.08, 1.05 ± 0.13 (F = 5.77, P < 0.05), and the mRNA levels of PAI-1 in aortic arch were 1.25 ± 0.11, 1.74 ± 0.06, 1.23 ± 0.05, 1.09 ± 0.14, 1.23 ± 0.04 (F = 23.56, P < 0.01).</p><p><b>CONCLUSION</b>The results indicate that TP seems to have regulatory function on transcription and protein levels of t-PA and PAI-1, in addition to maintaining the balance between PAI-1 and t-PA and healing the injury of fibrinolytic functions in rats induced by high-methionine dietary.</p>
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Animales , Masculino , Ratas , Dieta , Fibrinólisis , Metionina , Inhibidor 1 de Activador Plasminogénico , Sangre , Polifenoles , Farmacología , Ratas Wistar , Té , Química , Activador de Tejido Plasminógeno , SangreRESUMEN
<p><b>OBJECTIVE</b>To investigate the effects of garlic oil (GO) on n-hexane metabolized to 2, 5-hexanedione (2, 5-HD) in mice.</p><p><b>METHODS</b>Adult healthy Kunming-mice were treated with n-hexane and GO. The serum was obtained and extracted with ethyl acetate, and the levels of the serum 2, 5-HD were determined by gas chromatography.</p><p><b>RESULTS</b>(1) The concentration of 2, 5-HD in serum increased firstly after a single exposure to n-hexane (4 000 mg/kg). The peak value occurred at 10 hours after n-hexane treatment, but could hardly be detected at 20 h. (2) There was no 2, 5-HD in serum of control mice. The content of 2, 5-HD in serum increased along with the exposure dose of n-hexane. The serum 2, 5-HD contents of the 2000, 4000 and 6000 mg/kg groups mice were 8.04, 16.68 and 22.38 microg/ml at 8 h in pretreated mice, respectively, and showed significant dose-effect relationship. (3) When the different age mice were exposed to the same dose of n-hexane, the contents of 2, 5-HD in serum were significantly different after 8 hours (P<0.05). The serum 2, 5-HD level of the 5 weeks old mice (22.83 microg/ml) was much higher than the 4 (19.59 microg/ml) and 6 (16.42 microg/ml) weeks old mice. (4) When the different gender mice were exposed to the same dose of n-hexane, the concentration of 2, 5-HD in serum of female mice (13.22 microg/ml) was higher than that of the female mice (10.34 microg/ml, P<0.05). (5) GO significantly inhibited the increase of the serum 2, 5-HD levels of both the pretreatment and post-treatment groups treated with 80 mg/kg n-hexane respectively, but the pretreatment with GO exhibited the more suppressive effects than the post-treatment (P>0.05). Compared with the n-hexane group, the concentrations of serum 2, 5-HD in GO-pretreated groups mice decreased by 16.2%, 20.8%, 22.8% (P<0.05) and 32.1% (P<0.01), respectively, and showed significant dose-effect relationship.</p><p><b>CONCLUSION</b>The serum content of 2, 5-HD, the metabolite of n-hexane, is different in different genders and age mice after exposed to the same dose of n-hexane. GO can effectively inhibit the production of n-hexane metabolized to 2, 5-HD in mice serum.</p>
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Animales , Femenino , Masculino , Ratones , Compuestos Alílicos , Química , Biotransformación , Hexanos , Farmacocinética , Hexanonas , Sangre , Sulfuros , QuímicaRESUMEN
<p><b>OBJECTIVE</b>To investigate the dynamic changes of neurofilament contents in rat's spinal cord induced by 2, 5-hexanedione (2, 5-HD), and explore the molecular mechanism of n-hexane neuropathy.</p><p><b>METHODS</b>Male Wistar rats were administered at a dosage of 400 mg/kg/day 2, 5-HD for 2, 4 and 8 weeks respectively. HD-induced neurological defects were detected and quantified using gait score, and the relative lev-els of NF-H, NF-M, and NF-L in spinal cords of rats were determined by Western Blotting.</p><p><b>RESULTS</b>Exposure to 2, 5-HD produced progressive gait abnormalities, which suggested that the rat model of 2, 5-HD-induced neurotoxicity was established successfully. Western-Blotting results showed that NFs content in spinal cord demonstrated a progressive decline as the intoxication continued. In the supernatant fraction, compared to the controls, NF-H con-tent decreased by 15.7%, 57.0%, and 58.0% respectively after 2, 4, and 8-week treatment with 2, 5-HD (P < 0.01); accordingly, NF-M decreased by 36.0%, 61.3%, and 65.2% respectively (P < 0.01); NF-L decreased by 20.8%, 43.9%, and 44.3% respectively (P < 0.01). In the pellet fraction, the contents of NF-H in groups of 4 and 8 weeks' exposure to HD decreased by 35.6% and 43.2%, respectively (P < 0.01), and those of NF-L decreased by 26.4% and 42.1%, respectively (P < 0.01) when compared to the control. Further-more, NF-M contents in groups of 2, 4 and 8 weeks' exposure decreased by 23.3%, 33.9%, and 63.7% respectively (P < 0.01). The NFs level in spinal cords was highly correlated with gait abnormality of treated rats as the intoxication went on. Multiple correlation coefficients of NF-H, NF-M, and NF-L content with gait score of HD-treated rat were 0.8912, 0.9282 and 0.8981 (P < 0.01) respectively.</p><p><b>CONCLUSION</b>The declines of NFs are high-ly related to neurobehavioral abnormality of 2, 5-HD-treated animals, and involved in the development of n-hexane neuropathy.</p>
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Animales , Masculino , Ratas , Modelos Animales de Enfermedad , Marcha , Hexanonas , Toxicidad , Proteínas de Neurofilamentos , Metabolismo , Ratas Wistar , Médula Espinal , MetabolismoRESUMEN
<p><b>OBJECTIVE</b>To investigate whether the alterations of microtubule and microfilament expression are responsible for the neurotoxicity of carbon disulfide.</p><p><b>METHODS</b>Wistar rats were administered with carbon disulfide by gavage at a dosage of 300 or 500 mg/kg for continuous 12 weeks (five times per week). Spinal cords of carbon disulfide-intoxicated rats and their age-matched controls were Triton-extracted and ultracentrifuged to yield a pellet and a corresponding supernatant fraction. Then, the contents of alpha-tubulin, beta-tubulin, and beta-actin in both fractions were determined by immunoblotting. In the meantime, their mRNA levels in spinal cords were quantified using reverse transcriptase-polymerase chain reaction (RT-PCR).</p><p><b>RESULTS</b>In the supernatant fraction, the contents of beta-tubulin and beta-actin in both treated groups increased significantly (P < 0.01) the content of beta-tubulin increased by 141% and 158% respectively, and the content of beta-actin increased by 19% and 32% respectively. In the pellet fraction, the content of beta-tubulin in both groups increased by 107%(P < 0.01) and 118%(P < 0.01) respectively, and the others keep unaffected. In the meantime, the levels of of mRNA expression of beta-tubulin and beta-actin gene were elevated consistently in CS(2)-treated groups (P < 0.01) the levels of mRNA expression of beta-tubulin increased by 207% and 212% respectively, and the levels of mRNA expression of beta-actin increased by 94% and 91% respectively.</p><p><b>CONCLUSION</b>Carbon disulfide intoxication results in alternations of microtubule and microfilament expression, and the alternations might be related to its neurotoxicity.</p>
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Animales , Masculino , Ratas , Actinas , Genética , Metabolismo , Disulfuro de Carbono , Intoxicación , Modelos Animales de Enfermedad , ARN Mensajero , Genética , Ratas Wistar , Médula Espinal , Metabolismo , Tubulina (Proteína) , Genética , MetabolismoRESUMEN
<p><b>OBJECTIVE</b>To investigate the effect of carbon disulfide (CS(2)) on oxidation-antioxidation function of rat nerve tissues.</p><p><b>METHODS</b>Thirty male Wistar rats were randomly divided into the control group, the low-dosage exposure group and the high-dosage group, 10 rats each. The rats of the two exposure groups were administered with CS(2) by gavage at a dosage of 300 or 500 mgxkg(-1)xd(-1), 5 times every week for continuous 12 weeks. The alterations in glutathione (GSH), malondialdehyde (MDA), reactive oxygen species (ROS), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), hydrogen peroxidase (CAT) and total anti-oxidation (T-AOC) in cerebrum, spinal cord, and sciatic nerve of CS(2)-treated animals were assayed.</p><p><b>RESULTS</b>The results showed that the contents of MDA and ROS in nerve tissues of CS(2)-treated groups increased significantly except ROS in spinal cord and sciatic nerve of low dose group. The content of MDA was increased by 20.7% and 33.6% respectively in the cerebrum of the rats of the low-dosage group and the high-dosage group, by 18.5% and 23.3% respectively in the spinal cord, and by 20.7% and 53.0% respectively in the sciatic nerve, The content of MOS was increased by 20.1% and 34.9% respectively in the cerebrum of the rats of the low-dosage group and the high-dosage group, and by 14.1% and 15.4% respectively in the spinal cord and the sciatic nerve of the rats of the high-dosage group (P < 0.05 or P < 0.01). Furthermore, the activities of SOD, GSH-Px, CAT and T-AOC decreased significantly except GSH-Px and SOD in cerebrum of low dose group. The content of GSH was decreased by 17.2% and 26.5% respectively in the cerebrum of the rats of the low-dosage group and the high-dosage group, by 26.4% and 31.2% respectively in the spinal cord, and by 15.1% and 20.0% respectively in the sciatic nerve. The content of T-AOC was decreased by 11.1 and 26.4% respectively in the cerebrum of the rats of the low-dosage group and the high-dosage group, by 15.1% and 38.4% respectively in the spinal cord, and by 35.6% and 42.3% respectively in the sciatic nerve. The activity of SOD was decreased by 12.1% and 25.4% respectively in the spinal cord of the rats of the low-dosage group and the high-dosage group and by 16.4% and 30.3% respectively in the sciatic nerve. The activity of GSH-Px was decreased by 17.3% and 32.5% respectively in the spinal cord of the rats of the low-dosage group and the high-dosage group and by 17.1% and 21.5% respectively in the sciatic nerve. The activity of GSH-Px and SOD was decreased by 12.6% and 30.1% respectively in the cerebrum of the rats of the high-dosage group. The activity of CAT was decreased by 17.5% and 39.4% respectively in the cerebrum of the rats of the low-dosage group and the high-dosage group, by 25.2% and 31.3% respectively in the spinal cord, and by 17.1% and 36.9% respectively in the sciatic nerve (P < 0.05 or P < 0.01).</p><p><b>CONCLUSION</b>Subchronic exposure to CS(2) can induce significant changes of oxidation-antioxidation function in rat nerve tissues, which might be related to CS(2)-induced neurotoxicity.</p>
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Animales , Ratas , Antioxidantes , Metabolismo , Disulfuro de Carbono , Peroxidación de Lípido , Tejido Nervioso , Metabolismo , Ratas WistarRESUMEN
<p><b>OBJECTIVE</b>To investigate the dynamic changes of alpha-tubulin, beta-tubulin and beta-actin in sciatic nerve of hen with organophosphorus ester-induced delayed neuropathy (OPIDN).</p><p><b>METHODS</b>OPIDN was induced in 10-month-old Roman hens by daily subcutaneous administration of 30 mg/kg methamidophos for 15 days. Hens were sacrificed 2, 10, and 23 days respectively after manifesting neuropathy. The sciatic nerves were dissected, homogenized and used for the determination of the alpha-tubulin, beta-tubulin and beta-actin levels by western blotting.</p><p><b>RESULTS</b>The levels of alpha-tubulin in supernatant of sciatic nerves were decreased by 6%, 15% and 25% respectively on day 2, 10 and 23 respectively, while those in pellet remained almost unchanged. beta-tubulin were decreased by 27%, 6%, 19% in pellet and 1%, 21%, 22% in supernatant of sciatic nerves on 2, 10 and 23 days. Beta-actin level in pellet of sciatic nerve increased by 24%, 48% and 17% on day 2, 10 and 23, and little changes were observed in supernatant.</p><p><b>CONCLUSION</b>Methamidophos may induced changes of alpha-tubulin, beta-tubulin and beta-actin levels in sciatic nerve of hen, which may be one of the mechanism of the contribution to the occurrence and development of OPIDN.</p>
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Animales , Femenino , Actinas , Metabolismo , Pollos , Insecticidas , Toxicidad , Compuestos Organotiofosforados , Toxicidad , Nervio Ciático , Metabolismo , Tubulina (Proteína) , MetabolismoRESUMEN
<p><b>OBJECTIVE</b>To investigate the dynamic alterations of neurofilament subunits (NF) in sciatic nerve of hens with organophosphorus ester induced the delayed neurotoxicity or neuropathy (OPIDN).</p><p><b>METHODS</b>Hens with OPIDN were produced by giving 30 mg/kg methamidophos subcutaneously to the 10-month-old Roman hens daily for 15 days, and sacrificed after manifesting neurotoxic clinical signs on the 2nd, 10th, and 23rd day respectively. The sciatic nerves were dissected, homogenized and centrifuged. The levels of NF in supernatant and pellet of sciatic nerves were examined by Western blotting respectively at different time from 2 to 23 days.</p><p><b>RESULTS</b>Integrated optional density (IOD) of high molecular weight neurofilament (NF-H) in sciatic nerve pellet of hens on the day 2, 10, 23 after appearance of OPIDN were 145,117 +/- 17,038, 55,917 +/- 17,333 and 45,038 +/- 6,662 respectively. As compared with the control group (78,875 +/- 22,569), the contents of NF-H in pellet were increased by 84% on day 2, and decreased by 29% and 43% on day 10 and 23 respectively. IOD of NF-H in supernatant of sciatic nerves were 4,709 +/- 1,739, 12,337 +/- 3,205 and 16,745 +/- 931, which were reduced significantly as compared with the control (44,083 +/- 6,895) at three different times. There was no significant difference in IOD of middle molecular weight neurofilament (NF-M) between control group (27,925 +/- 2,660) and on day 2 (31,493 +/- 4,625) in pellet. Those were 19,367 +/- 2,746 and 6,612 +/- 1,119 respectively on day 10 and day 23 in pellet of hen's sciatic nerve, which were much less than that in control. Little were detected in supernatant on day 10, and the IOD of NF-M were 3,196 +/- 269 and 5,206 +/- 1,292 on day 2 and day 23 respectively, which were lessened by 81% and 70% as compared with the control (17,243 +/- 3,232). In sciatic nerve pellet of hens, IOD of low molecular weight neurofilament (NF-L) on day 2 was 39,211 +/- 3,800, which was much higher than that in the control (28,749 +/- 9,319). There were no significant differences between IOD on day 10 (27,974 +/- 3,611), day 23 (21,507 +/- 2,286) and the control. There was no detection both on day 2 and 10 in supernatant of sciatic nerve, and IOD of NF-L were 5,962 +/- 1,929 on day 23, which were reduced significantly compared with the control (11,897 +/- 352).</p><p><b>CONCLUSION</b>The alterations of NF in sciatic nerve might contribute to the occurrence and development of OPIDN.</p>
Asunto(s)
Animales , Femenino , Pollos , Insecticidas , Toxicidad , Proteínas de Neurofilamentos , Metabolismo , Compuestos Organotiofosforados , Toxicidad , Nervio Ciático , Metabolismo , Patología , Pruebas de ToxicidadRESUMEN
<p><b>OBJECTIVE</b>To investigate the effect of ginsenoside-Rg3 on lung metastasis of ribonuclease inhibitor (RI) gene-transfected mouse B16 melanoma.</p><p><b>METHODS</b>C57BL/6 mice were iv injected with parental or RI-transfected B16 melanoma cells. Lung metastasis was assessed by the number of surface tumor nodules. Mice were divided into 6 groups. Group I, II and III of mice were given parental, mock-transfected and RI-transfected B16 melanoma cells, respectively while in group IV, V and VI, Rg3 (1.5 mg/kg, iv q.o.d. x 10) was given to mice bearing parental, mock-transfected and RI-transfected B16 melanoma, respectively. Micovessel density (MVD) of the lung metastatic tumor was assessed by immunohistochemical staining of factor VIII-R expression.</p><p><b>RESULTS</b>The number of tumor nodules was significantly decreased in mice injected with RI-transfected B16 melanoma (Gp III, compared to Gp I and II). Rg3 treatment per se could also decrease the number of lung tumor nodules but to a lesser extent (Gp IV and V compared to Gp III). However, Rg3 synergized with RI transfection resulting in most significant inhibition of lung metastasis (Gp VI). Mice in Gp I and II died within 26 days of the experiment, whereas all the mice in Gp VI were alive during the observation period of one and one half month. MVD was significantly decreased in the lung tumor nodules in mice injected with RI-transfected B16 melanoma. It was further decreased when additional Rg3 was given (Gp VI).</p><p><b>CONCLUSION</b>Transfection of ribonuclease inhibitor gene significantly reduces the metastatic potential of B16 melanoma. Ginsenoside-Rg3 has a synergistic effect.</p>
Asunto(s)
Animales , Masculino , Ratones , Antineoplásicos Fitogénicos , Farmacología , Línea Celular Tumoral , Ginsenósidos , Farmacología , Neoplasias Pulmonares , Patología , Melanoma Experimental , Genética , Patología , Ratones Endogámicos C57BL , Trasplante de Neoplasias , Panax , Química , Hormonas Placentarias , Genética , TransfecciónRESUMEN
<p><b>OBJECTIVE</b>To establish the value of the resting energy expenditure (REE) in healthy newborns and evaluate relative factors of REE.</p><p><b>METHODS</b>One hundred and fifty-four healthy newborns (75 boys, 79 girls; birth-weight 2,500-3,999 g) were enrolled in this study. The Apgar score at the 5th minute was equal to or more than 8; the postnatal age was equal to or more than 5 days. The newborns had no apparent defect. The mothers had no history of metabolic and endocrine diseases. REE was measured by Deltatrac II in child canopy mode for 30-45 minutes during asleep or quiet awake status.</p><p><b>RESULTS</b>The average REE was (201.8 +/- 25.4) kJ/(kg.d), which was significantly lower than the predicted REE by Schofield formula[(226.1 +/- 4.8) kJ/(kg.d), P = 0.000], the predicted REE was 12.04% higher than the measured REE. There were no differences in sex and ways of delivery. The newborns whose birth-weight was between 2,500-2,999 g were measured in two modes: baby and child mode, and the REE values were significantly higher (122.6 +/- 25.0) kJ/(kg.d) and (208.8 +/- 26.4) kJ/(kg.d), respectively (P = 0.000).</p><p><b>CONCLUSIONS</b>The prediction formula might be improper for calculating the REE in newborn infants. The indirect calorimetry was the better way to know the actual REE of newborns. The authors recommended that in child mode the measurement of REE in newborns would be the indirect calorimetry, and the REE in healthy newborns was (201.8 +/- 25.4) kJ/(kg.d).</p>