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Aim To explore the effect of γ-ray on the mRNA,protein expression levels and metabolic activity level of the key drug metabolic enzyme CYP3A1 in rat liver. Methods Wistar rats were randomly divided into control group, 24 h post-radiation group and 72 h post-radiation group. The experimental group was exposed to total body irradiation of single 6 Gy γ-ray. Blood was collected from the orbital venous plexus for blood routine examination and biochemical analysis 24 h and 72 h after irradiation, and liver tissue was prepared for quantifying expression of CYP3A1 mRNA and liver-specific microRNA (miR-122-5p) through RT-PCR. The expression level of CYP3A1 protein was analyzed by Western blot, and the metabolic activity level of CYP3A1 detected by the specific substrate midazolam combined with LC-MS method. Results Com¬pared with the control group, the weights of the rats in the radiation group significantly decreased, and the number of white blood cells was markedly reduced. Simultaneously, the activities of alanine aminotrans-ferase and alkaline phosphatase continuously descended, as well as the levels of total bilirubin and bile acid significantly increased, which indicated that the liver may be damaged after radiation. The relative expression of CYP3A1 mRNA continued to increase significantly 24 h and 72 h after irradiation. CYP3A1 protein expression and metabolic activity levels showed an obvious increasing trend 24 h after irradiation, and rose significantly 72 h after irradiation compared with the control group. At the same time, the expression of miR-122-5p in liver of rats in the 24 h and 72 h post-radiation group continued to decrease rapidly compared with the control group. Conclusions γ-ray radiation may arouse damage effect on liver, which leads to the continuous up-regulation of the mRNA and protein expression levels of the capital metabolic enzyme CYP3A1 in liver tissue, as well as the elevation of the metabolic activity level. The regulatory mechanism might be related to miR-122-5p.
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Recombinant humanized anti-ricin monoclonal antibody (MIL50) is a recombinant humanized monoclonal antibody targeting ricin. In this study, an ELISA method was used to establish a method for the determination of MIL50 in macaque serum, and a cross design method was used. Twelve rhesus monkeys were intravenously injected 1 mg·kg-1 test preparation (MIL50 freeze-died powder injection) and reference preparation (MIL50 liquid preparation) to determine the plasma concentration of MIL50 at different time points, and the pharmacokinetic parameters were analyzed to compare the pharmacokinetic characteristics of MIL50 liquid preparation and freeze-died powder injection in rhesus monkeys. Animal welfare and experimental procedures follow the regulations of the Animal Ethics Committee of the Chinese Academy of Medical Sciences and Use of Laboratory Animals and the regulations derived by the Animal Care and Welfare Committee of the Institute of Radiation Medicine, Academy of Military Medical Sciences (IACUC-DWZX-2020-503). The results showed that there was no significant difference between Cmax and AUC0-5d in the two groups. The liquid preparation was the reference preparation, with Cmax ratio of 101.6% and AUC0-5d ratio of 101.9%, the 90% confidence interval of Cmax was 79.42%-129.92%, and the 90% confidence interval of AUC0-5d was 85.72%-121.18%. These results suggested that different dosage forms of MIL50 had certain differences in the changes of blood drug concentration in rhesus monkeys.
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BACKGROUND@#Hyperuricemia and gout have become public health concerns; many important guidelines have recommended xanthine oxidase inhibitors (XOIs) as the first-line urate-lowering therapies (ULTs) to treat chronic gout with hyperuricemia. However, whether treating hyperuricemia and gout with ULTs modifies cardiovascular risks remains controversial. The aim of this study was to assess the incident risk of cardiovascular (CV) events (CVE) in hyperuricemia population, assess the cardiovascular benefit-risk of ULTs in hyperuricemia patients with or without gout in diverse cardiovascular risk sub-groups, and specify the safety of different ULTs.@*METHODS@#We searched PubMed, Embase, the Cochrane Library, Wanfang, Chongqing VIP (CQVIP, en.cqvip.com), and China National Knowledge Infrastructure Database for prospective cohort studies and randomized controlled trials (RCTs) in English and Chinese. Potential medications included XOIs, and uricosurics. RCTs were divided into sub-groups analysis based on blinding status and patients' history of CV diseases. Risk ratios (RRs) were calculated and were reported with corresponding 95% confidence intervals (CIs) by fixed-effects or random-effects model.@*RESULTS@#Seven prospective cohort studies and 17 RCT studies were included. The risks of both major adverse cardiovascular events (MACE) (RR = 1.72, 95% CI 1.28-2.33) and CVE (RR = 1.35, 95% CI 1.12-1.62) were higher in the hyperuricemia population than non-hyperuricemia one. In seven RCT studies where XOIs were compared with no-treatment or placebo, the results of five low CV risk studies showed that XOIs lowered the risks of both MACE (RR = 0.35, 95% CI 0.20-0.62) and CVE (RR = 0.61, 95% CI 0.44-0.85); whereas two high CV risk studies showed that XOIs lowered the risk of CVE (RR = 0.69, 95% CI 0.54-0.88) rather than MACE (RR = 0.62, 95% CI 0.29-1.35). In nine RCT studies where the cardiovascular safety between febuxostat and allopurinol were compared, no statistical difference was found in the risk of MACE or CVE.@*CONCLUSIONS@#The hyperuricemia population does have a higher incidence of CVE, and the results suggested that XOIs might reduce the incidence of MACE and total CVE. In addition, from the perspective of cardiovascular safety, febuxostat equaled allopurinol in our meta-analysis.
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Objective To discuss the effects of oxygen therapy on partial pressure of oxygen, pulmonary function and quality of life among patients with chronic obstructive pulmonary disease (COPD) during respiratory rehabilitation (PR) . Methods Eighty patients with COPD were selected and randomly divided into two groups (receiving oxygen therapy and receiving no oxygen therapy) with 40 in each group.Patients who participated in this study underwent a comprehensive 8-week outpatient PR program. After the 8-week treatment, patients'pulmonary function, blood gas analysis and St.George respiratory questionnaire (SCRQ) score were tested. Results Significant improvements were observed in pulmonary function, partial pressure of oxygen and SCRQ score in both groups after the treatment.The group underwent the oxygen therapy showed better results in partial pressure of oxygen, FEV1 and SCRQ score during the rehabilitation compared to the group underwent no oxygen therapy. Conclusion Oxygen therapy during PR plays a positive role in respiratory rehabilitation, which is worth generalizing.
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As a widespread phenomenon in living system, molecular self-assembly has become the meeting point of multidisciplinary research including chemistry, biology, materials science and medicine. In recent years, the rapid development in molecular self-assembly of peptide technology is showing a great potential in the application of tissue engineering, drug delivery, bionic medicine, cosmetology field, optical and electronic product development, etc. Especially, the remarkable hemostatic effect of self-assembling peptides (SAP) on organs, nerves and brain wounds successfully promoted its application to the material science and clinical medicine. This review focuses on the hemostatic effects and characteristics of SAP on different bleeding wound models, action mechanism, its benefits and limitations as well as its adrancing trends.
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Humanos , Sistemas de Liberación de Medicamentos , Hemostasis , Péptidos , Ingeniería de TejidosRESUMEN
<p><b>OBJECTIVE</b>To evaluate the short-term efficacy and safety of Bushen Shuji Granule (BSG) in treating ankylosing spondylitis (AS) patients.</p><p><b>METHODS</b>A prospective randomized controlled clinical trial was carried out in 62 active stage AS patients with Shen deficiency Du-channel cold syndrome (SDDCS), who were randomly assigned to the BSG group (treated with BSG) and the control group (treated with Celecoxib Capsule). Twelve weeks consisted of one therapeutic course. Therapeutic effects were evaluated by ASAS20 and ASAS40 (set by Assessments in Ankylosing Spondylitis working group) , BASDA150, Chinese medical (CM) syndrome efficacy evaluation standards. BASDAI, the Bath Ankylosing Spondylitis Functional Index (BASFI), the Bath AS Metrology Index (BASMI), scores for spine pain, scores for pain at night, patient global assessment (PGA) , erythrocyte sedimentation rate (ESR) , and C reactive protein (CRP) were observed before and after treatment.</p><p><b>RESULTS</b>After three-month treatment by BSG, ASAS20 standard rate was 63. 33% (19/30 cases) in the BSG group and 66.67% (20/30 cases) in the control group with no significant difference between the two groups (χ2 = 0.073, P > 0.05). The efficacy for CM syndromes was 70.00% (21/30 cases) in the BSG group, higher than that in the control group [40.00% (12/30 cases), χ2 = 5.455, P < 0.05]. Scores for CM syndromes, BASDAI, night pain index, spinal pain index, PGA, CRP were improved in the BSG group (P < 0.05, P < 0.01). The incidence of adverse events in the BSG group was lower than that of the control group.</p><p><b>CONCLUSION</b>BSG based on Shen supplementing, Du-channel strengthening, blood activating, and channels dredging method had good short-term clinical efficacy and safety in treating AS.</p>
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Humanos , Pueblo Asiatico , Investigación Biomédica , Sedimentación Sanguínea , Proteína C-Reactiva , Medicamentos Herbarios Chinos , Usos Terapéuticos , Dolor , Estudios Prospectivos , Seguridad , Espondilitis Anquilosante , QuimioterapiaRESUMEN
A rapid, sensitive and simple liquid chromatography-tandem mass spectrometric (LC-MS/MS) method was developed for the simultaneous determination of yogliptin and its metabolite in Wistar rat plasma. Linagliptin and dexamethasone were chosen as the internal standards of yogliptin and its metabolite, (R)-8-(3-hydroxypiperidine- -yl)-7-(but-2-yn-1-yl)-1-((5-fluorobenzo[d]thiazol-2-yl)methyl)-3-methyl- H-purine-2, 6 (3H, 7H)-dione, respectively. After a simple protein precipitation using acetonitrile as the precipitating solvent, both analytes and ISs were separated on a Grace Altima HP C18 column (2.1 mm x 50 mm, 5 microm) with gradient elution using methanol (containing 0.1% formic acid, 4 mmol x L(-1) ammonium acetate)-0.1% formic acid (containing 4 mmol x L(-1) ammonium acetate) as the mobile phase. A chromatographic total run time of 4.4 min was achieved. Mass spectrometric detection was conducted with electrospray ionization under positive-ion and multiple-reaction monitoring modes. Linear calibration curves for yogliptin and its metabolite were over the concentration range of 0.5 to 500 ng x mL(-1) with a lower limit of quantification of 0.5 ng x mL(-1). The intra- and inter- assay precisions were all below 14%, the accuracies were all in standard ranges. The method was used to determine the concentration of yogliptin and M1 in Wistar rat plasma after a single oral administration of yogliptin (27 mg x kg(-1)). The method was proved to be selective, sensitive and suitable for pharmacokinetic study of yogliptin and M1 in Wistar rat plasma.
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Animales , Ratas , Cromatografía Liquida , Dexametasona , Sangre , Inhibidores de la Dipeptidil-Peptidasa IV , Sangre , Farmacocinética , Linagliptina , Sangre , Ratas Wistar , Espectrometría de Masas en TándemRESUMEN
This paper is to report the study of the metabolism of forscolin in plasma and liver microsomes for guiding clinical therapy. Forscolin was quantified by HPLC-MS/MS. The metabolic stability of forscolin in rat, Beagle dog, monkey and human plasma and liver microsomes, mediated enzymes of forscolin and its inhibition on cytochrome P450 isoforms in human liver microsomes were studied. Results showed that forscolin was not metabolized in plasma of the four species but metabolized in liver microsomes of the four species. The t1/2 of forscolin in rat, Beagle dog, monkey and human liver microsomes were (52.0 +/- 15.0), (51.2 +/- 5.9), (6.0 +/- 0.2) and (11.9 +/- 1.8) min; CL(int) were (75.6 +/- 18.7), (60.9 +/- 6.8), (513.8 +/- 14.3) and (176.2 +/- 25.6) mL x min(-1) x kg(-1); CL were (34.8 +/- 4.5), (23.3 +/- 1.0), (40.3 +/- 0.5) and (17.9 +/- 0.3) mL x min(-1) x kg(-1), respectively. Forscolin was metabolized by CYP3A4 in human liver microsomes. There was definite inhibition on CYP3A4 at the concentrations of forscolin between 0.1 ng x mL(-1) and 5 microg x mL(-1). Therefore, forscolin is rapidly excreted from liver microsomes. Attention should be paid to the drug interaction when forscolin was used along with other drugs metabolized by CYP3A4 in clinics.
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Animales , Perros , Humanos , Ratas , Cromatografía Líquida de Alta Presión , Coleus , Química , Colforsina , Sangre , Metabolismo , Citocromo P-450 CYP3A , Metabolismo , Macaca , Tasa de Depuración Metabólica , Microsomas Hepáticos , Metabolismo , Plantas Medicinales , Química , Espectrometría de Masas en TándemRESUMEN
Healthy Beagle dogs were administrated with batroxobin by intravenous infusion at high, medium and low doses. The study of pharmacodynamics and pharmacokinetics was intended to clarify the relevance of them and provided strong evidence for clinical use of batroxobin. The blood samples were collected after injection based on the time schedule and samples were tested by ELISA method to get the concentration of batroxobin. At the same time, changes of prothrombin time (PT), thrombin time (TT), activated partial thromboplastin time (APTT), fibrinogen (Fib) and D-dimmer were tested. The results showed that the concentration of D-D increased significantly after administration compared with that of before administration. The main pharmacokinetic parameters were as follows: t1/2 were (2.27 +/- 0.42) h, (10.65 +/- 2.19) h and (11.01 +/- 3.51) h; C(max) were (11.9 +/- 1.72) ng x mL(-1), (154.53 +/- 12.38) ng x mL(-1) and (172.14 +/- 47.33) ng x mL(-1); AUC(last) were (29.38 +/- 3.69) ng xh x mL(-1), (148.43 +/- 72.85) ng x h x mL(-1) and (599.22 +/- 359.61) ng x h x mL(-1). The elimination of batroxobin was found to be in accord with linear kinetics characteristics. The results of pharmacodynamics showed that D-dimmer level increased significantly after the administration of batroxobin, which was similar with the changes of batroxobin plasma concentration. Simultaneously, Fib concentrations in Beagle dog blood decreased significantly after the iv administration of batroxobin, while recovered to base level after 48 hours. PT, TT and APTT significantly became longer after administration, which returned to normal level after 48 hours. Especially, the D-dimmer levels and the batroxobin concentration in plasma after intravenous infusion of the drug were synchronized in Beagle dogs. Changes between PD/PK results had obvious correlation, and the D-dimmer levels in plasma can be one of the important monitoring indicators of batroxobin in thrombolytic medication.
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Animales , Perros , Masculino , Área Bajo la Curva , Batroxobina , Sangre , Farmacocinética , Farmacología , Ensayo de Inmunoadsorción Enzimática , Productos de Degradación de Fibrina-Fibrinógeno , Metabolismo , Fibrinógeno , Metabolismo , Fibrinolíticos , Sangre , Farmacocinética , Farmacología , Infusiones Intravenosas , Tiempo de Tromboplastina Parcial , Tiempo de Protrombina , Tiempo de TrombinaRESUMEN
This paper is to report the study of the metabolism of lidamycin in vitro including in plasma and microsomes to guide clinical therapy. Lidamycin was quantified by detecting its active ingredient using HPLC-MS/MS. The metabolic stability of lidamycin in rat, Beagle dog, monkey and human plasma and liver microsomes, and its inhibition to cytochrome P450 isoforms in human liver microsomes were studied. Results showed that lidamycin was metabolized in the four species of plasma, and the sequence of metabolic rates in plasma were in rat > in dog > in human > in monkey. But among the four species of liver microsomes, lidamycin was metabolized only in monkey liver microsomes. There was almost no inhibition to cytochrome P450 isoforms at the concentrations of between 0.0005 and 10 ng x mL(-1). Therefore, the property of lidamycin metabolism in human is similar with that in dog, and metabolism of other drugs would not be decreased by cytochrome P450 as used along with lidamycin in clinic.
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Animales , Perros , Humanos , Ratas , Aminoglicósidos , Sangre , Metabolismo , Antibióticos Antineoplásicos , Sangre , Metabolismo , Cromatografía Líquida de Alta Presión , Citocromo P-450 CYP1A2 , Metabolismo , Sistema Enzimático del Citocromo P-450 , Metabolismo , Enediinos , Sangre , Metabolismo , Activación Enzimática , Macaca , Microsomas Hepáticos , Metabolismo , Espectrometría de Masas en TándemRESUMEN
Objective To explore the iodine level in the environment and the iodine status among the general population as well as the prevalence of thyroid nodules in Hangzhou city.Relationship between the prevalence of thyroid nodules and the policy of universal salt iodization in Hangzhou was also analyzed.Methods Questionnaire,a 3- day weighed dietary record method,and 3 days' 24- hour dietary recall method were used to understand the iodine nutrition status and dietary intake of iodine among the general population in the city.Drinking water,edible salt and morning urine were collected to determine iodine content.All objects under survey underwent the thyroid B ultrasonic examination.Statistical analysis was done by SPSS 13.0 and SAS 9.1.Results (1)In total,12 620 effective questionnaires were available,with 221 water samples,12 730 urine samples,and 3593 salt samples collected.12 515 objects underwent B ultrasonic examination,and 1848received dietary investigation.(2)Water iodine level of Hangzhou was in the range of 0.20-5.99 μg/L,with the median level as 2.58 μg/L.(3) Average daily dietary intake of iodine for adult males in Hangzhou was 289.2 lμg/d.The contribution of iodine intake from iodized salt was 74.4%.(4) The median of Hangzhou residents' urinary iodine was 178.80 μg/L,with the urinary iodine levels at 100 μg/L-,200 μg/L-,<100 μg/L,and ≥300 μg/L groups were 37.14%,23.11%,21.05%,and 18.69% respectively.Urinary iodine of pregnant women was 141.0 tg/L.(5) Incidence of thyroid nodules in females(28.6% ) was higher than that of males(20.1% ).The detection rate increased with age (6.4% at group 6-,10.9% at 12-,12.0% at 18-,24.4% at 40-,and 38.8% at 65-) ; with the highest in urban area (29.8%),followed by suburbs (23.3%) and in rural area it showed the least (20.3%).Urinary iodine level was found lower among the population who had been detected with thyroid nodules (160.36 μ g/L) than those among the undetected population (182.00 μg/L).Conclusion Hangzhou appeared to be an area where the environmental was iodine deficient.Iodized salt was the major source of iodine intake.The iodine status among the general population seemed to be safe and suitable,but the iodine level for pregnant women was not sufficient.There was still no evidence indicating that the universal salt iodization policy in Hangzhou was associated with the prevalence of thyroid nodules.
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<p><b>OBJECTIVE</b>To explore the correlation of the expressions of metastatic tumor antigen 1 (MTA1) and hypoxia-inducible-factor l alpha (HIF-1alpha) to the clinical features of lung cancer.</p><p><b>METHODS</b>The expressions of MTA1 and HIF-1alpha proteins in 59 lung cancer patients were detected by immunohistochemistry.</p><p><b>RESULTS</b>Positive staining of MTA1 and HIF-1alpha was found in 54.24% and 50.85% of the patients, respectively. MTA1 expression was not found to correlated to any of the clinical parameters. HIF-1alpha expression was significantly lower in poorly differentiated than in moderately and well differentiated lung cancer (P<0.05), and a positive association between the expressions of MTA1 and HIF-17alpha were noted (P<0.05).</p><p><b>CONCLUSION</b>MTA1 and HIF-1alpha overexpression occurs in lung cancer possibly as an important factor of lung carcinogenesis. MTA1 may promote lung carcinogenesis by enhancing HIF-1alpha protein activity.</p>
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Adulto , Anciano , Femenino , Humanos , Masculino , Persona de Mediana Edad , Regulación Neoplásica de la Expresión Génica , Histona Desacetilasas , Metabolismo , Subunidad alfa del Factor 1 Inducible por Hipoxia , Metabolismo , Inmunohistoquímica , Neoplasias Pulmonares , Genética , Metabolismo , Patología , Proteínas Represoras , MetabolismoRESUMEN
The paper is to report the pharmacokinetic character of a series of chemical compounds in vitro and in vivo. Metabolism stability of a series of chemical compounds was screened by using rat liver microsomes. The samples of different chemical compounds were combined and then simultaneously detected by LC-MS/MS. Compounds y13, y12 and y11 were screened out by microstability assay in vitro. The pharmacokinetics of compounds y11, y12 and y13 was evaluated by using SD rat. The plasma samples were pooled at the same time. The plasma concentrations were determined by LC-MS/MS. The pharmacokinetic character of two compounds y13, y11 was good by screening in vivo, so they were developed for further research. High-throughput screening of drug candidates in vitro and in vivo was effective, to provide information for the chemical structure information and lower the drug development risk.
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Animales , Femenino , Masculino , Ratas , Cromatografía Liquida , Métodos , Evaluación Preclínica de Medicamentos , Métodos , Ensayos Analíticos de Alto Rendimiento , Métodos , Microsomas Hepáticos , Metabolismo , Preparaciones Farmacéuticas , Sangre , Metabolismo , Farmacocinética , Distribución Aleatoria , Ratas Sprague-Dawley , Espectrometría de Masa por Ionización de Electrospray , MétodosRESUMEN
the migrant workers' families.
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<p><b>OBJECTIVE</b>To study the pathological basis of radiological reaction types of radiation-induced liver disease on multiphasic CT scans.</p><p><b>METHODS</b>Three pigs (tagged with A, B, and C) were subjected to single-dose radiation of 40, 40 and 30 Gy on the right or left lobe of the liver, respectively. At 42, 56, 133, and 168 days after irradiation, all pigs were examined with non-enhanced scan and contrast-enhanced scans at different time points after contrast injection. Hounsfield units (HU) were measured in each CT study to evaluate the density of irradiated and non-irradiated liver tissue to determine the reaction type. Liver tissues in the irradiation area obtained by needle biopsy with CT guidance were examined with electron microscopy, and specimens of the tissue corresponding to the region of interest on CT were obtained from necropsies for pathological examination.</p><p><b>RESULTS</b>Radiologically, the 3 pig models presented with 3 reaction types on the multiphasic CT scans on days 133, 56, and 168 after radiation, respectively. Type 1 presented constant low-density change in all phases, the pathological basis of which was radiation hepatitis; type 2 showed pre-contrast phase isodense, arterial phase hyperdense, portal phase isodense and later phase hyperdense changes; type 3 was characterized by pre-contrast phase isodense, arterial phase hyperdense, portal phase hypodense and later phase hyperdense changes. The pathological basis of the last two radiological reaction types was radiation cirrhosis (postnecrotic cirrhosis).</p><p><b>CONCLUSIONS</b>Different radiological reaction types of radiation liver injury on multiphase CT have different pathological basis, and multiphase contrast-enhanced CT may help distinguish the radiation reactions from tumor recurrence.</p>
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Animales , Femenino , Masculino , Hepatopatías , Diagnóstico por Imagen , Patología , Proyectos Piloto , Traumatismos Experimentales por Radiación , Diagnóstico por Imagen , Patología , Coloración y Etiquetado , Porcinos , Tomografía Computarizada por Rayos XRESUMEN
<p><b>OBJECTIVE</b>To review the appearance of radiation-induced liver injury on computer tomography for quantitative assessment of dosimetric changes in different radiological reactions and the influence of time-effect.</p><p><b>METHODS</b>The focal liver reactions of 35 patients treated with three-dimensional conformal radiation therapy (3-D CRT) for liver malignancies were evaluated, with the applied doses of 36-65 Gy in 4-28 fractions completed in 8-41 days. All patients received nonenhanced CT scan and arterial-dominant phase contrast-enhanced CT scan 1-6 months after therapy. The liver tissue density in irradiated and nonirradiated liver was compared, and the reaction type and the threshold dose determined radiologically.</p><p><b>RESULTS</b>On at least one follow-up examination, 51.4% of patients were found to have a focal radiation reaction in the liver. The radiation reaction was hypodense in 43.75% of the follow-up nonenhanced CT examinations and in 19.23% of arterial-dominant phase contrast-enhanced CT scans. It was hyperdense in 42.31% of arterial-dominant phase contrast-enhanced CT scans. The median threshold dose inducing a radiation reaction was 30.8 Gy (range 18-42.8 Gy). The detected threshold dose was positively correlated with the time of detection of the reaction (P=0.041), with a correlation coefficient of -0.473. On arterial-dominant phase contrast-enhanced CT scans, the threshold dose was significantly higher for hyperdense than for hypodense changes (P=0.017). In additional follow-up, the reaction volume decreased and the reaction types changed on arterial-dominant phase contrast-enhanced CT scans.</p><p><b>CONCLUSIONS</b>The threshold dose can be different in different radiological reaction types on multiphase CT scans. The detected threshold dose is inversely correlated with the time of detection of the early reaction. Multiphase contrast-enhanced CT is helpful to distinguish radiation reactions from recurrent tumors.</p>
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Adulto , Anciano , Femenino , Humanos , Masculino , Persona de Mediana Edad , Carcinoma Hepatocelular , Radioterapia , Hígado , Diagnóstico por Imagen , Patología , Efectos de la Radiación , Neoplasias Hepáticas , Radioterapia , Traumatismos por Radiación , Diagnóstico por Imagen , Dosificación Radioterapéutica , Radioterapia Conformacional , Métodos , Reproducibilidad de los Resultados , Tomografía Computarizada por Rayos X , MétodosRESUMEN
This study was purposed to verify the binding part of human complement C3 to complement receptor III (CRIII) in monocytes, the peptide rC3B, including the binding-site, was expressed, purified and identified. rC3B, the binding part of human complement C3 to CRIII, was selected by computer-aided modeling and summarizing researches published. Then, rC3B gene fragment was amplified by PCR, and cloned into prokaryotic vector pQE30a. The fusion protein rC3B was expressed in E.coli M15 and purified by Ni(2+)-chelating affinity chromatography. The activity of rC3B was identified by Western blot and adherence assay with monocytes. The results showed that rC3B fragment was obtained, and a prokaryotic expression vector pQE30-rC3B was constructed. rC3B was efficiently expressed and purified. In Western blot, the target protein showed the activity of binding with C3 antibody, while the purified protein showed the activity of adherence with monocytes. It is concluded that the recombinant C3B was obtained and identified, and this study lay the basis for the further functional analysis of C3.