RESUMEN

Severe acute pancreatitis (SAP) is a common acute abdominal disease accompanied by systemic inflammatory response syndrome, which may be complicated by acute kidney injury (AKI). Isoacteoside (ISO) is the active ingredient of Monochasma savatieri Franch. ex Maxim and has been reported to have anti­inflammatory activities. The present study detected the effects of ISO on AKI induced by SAP in rat models, and the underlying mechanism. The optimum dose of ISO for treatment of AKI induced by SAP was determined. The serum levels of TNF­α and IL­6 were estimated using an ELISA. Kidney injury was evaluated by histopathological examination, and the expression levels of nitric oxide were also detected. The expression levels of Toll­like receptor 4 (TLR4) and NF­κB p65 were measured by immunohistochemistry and western blotting. The results revealed that ISO may serve a critical role in ameliorating AKI induced by SAP. These effects may be associated with the TLR4/NF­κB signaling pathway.

Asunto(s)

Lesión Renal Aguda/prevención & control , Glucósidos/farmacología , Riñón/efectos de los fármacos , Pancreatitis/complicaciones , Fenoles/farmacología , Lesión Renal Aguda/sangre , Lesión Renal Aguda/etiología , Animales , Antiinflamatorios/farmacología , Interleucina-6/sangre , Riñón/metabolismo , Riñón/patología , Masculino , FN-kappa B/metabolismo , Óxido Nítrico/metabolismo , Pancreatitis/patología , Ratas Sprague-Dawley , Índice de Severidad de la Enfermedad , Transducción de Señal/efectos de los fármacos , Receptor Toll-Like 4/metabolismo , Factor de Transcripción ReIA/metabolismo , Factor de Necrosis Tumoral alfa/sangre

RESUMEN

BackgroundTo investigate the impact of goggles on their health and clinical practice during management of patients with COVID-19. Methods231 nurse practitioners were enrolled who worked in isolation region in designated hospitals to admit patients with COVID-19 in China. Demographic data, goggle-associated symptoms and underlying reasons, incidence of medical errors or exposures, the effects of fog in goggles on practice were all collected. Data were stratified and analyzed by age or working experience. Risk factors of goggle-associated medical errors were analyzed by multivariable logistical regression analysis. FindingsGoggle-associated symptoms and foggy goggles widely presented in nurses. The most common symptoms were headache, skin pressure injury and dizziness. Headache, vomit and nausea were significantly fewer reported in nurses with longer working experience while rash occurred higher in this group. The underlying reasons included tightness of goggles, unsuitable design and uncomfortable materials. The working status of nurses with more working experience was less impacted by goggles. 11.3% nurses occurred medical exposures in clinical practice while 19.5% nurses made medical errors on patients. The risk factors for medical errors were time interval before adapting to goggle-associated discomforts, adjusting goggles and headache. InterpretationGoggle-associated symptoms and fog can highly impact the working status and contribute to medical errors during management of COVID-19. Increased the experience with working in PPE through adequate training and psychological education may benefit for relieving some symptoms and improving working status. Improvement of goggle design during productive process was strongly suggested to reduce incidence of discomforts and medical errors.

RESUMEN

Objective::To observe the effect of modified Banxia Xiexintang on depression during perimenopause, in order to study itseffecton 5-hydroxytryptamine (5-HT) and proinflammatory factors. Method::One hundred and thirty-nine patients were randomly divided into control group (69 cases) and observation group (70 cases) by random number table.Patients in control group got tibolone tablets, 2.5 mg/time, 1 time/day, and paroxetine hydrochloride tablets, 20 mg/time, 1 time/day.In addition to the therapy in control group, patients in observation group were added with modified Banxia Xiexintang, 1 dose/day.The course of treatment was 8 weeks.And before and after treatment, Hamilton depression scale for-17 items (HAMD-17), Zung's self-rating depression scale (SDS), hamilton anxiety scale (HAMA), improvement Kupperman(KI), liver depression and spleen deficiency syndrome, menopause-specific quality of life questionnaire (MENQOL) and treatment emergent symptom scale (TESS) were scored, and levels of 5-HT, rain-derived neurotrophic factor (BDNF), interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) were detected. Result::After treatment, scores of HAMD-17 and SDS in observation group were lower than those in control group (P<0.01). And the effect on trea depression in observation group was better than that in control group (Z=2.074, P<0.05). The degree of depression in observation group was ligher than that in control group (Z=2.157, P<0.05). And scores of HAMA, KI and liver depression and spleen deficiency syndrome were lower than those in control group (P<0.01). The severity of perimenopausal syndrome was ligher than that in control group (Z=2.046, P<0.05). And scores of vasomotor symptoms and psychological symptoms of MENQOL scale and the total scores were lower than those in control group (P<0.05). Levels of 5-HT and BDNF were higher than those in control group (P<0.01), while levels of IL-1β, TNF-α and TESS were lower than those in control group (P<0.01). Conclusion::In addition to theroutine western medicine, modified Banxia Xiexintang can alleviate the severity of depression, release the symptoms of depression, anxiety and perimenopausal syudrome(PMS), improve the quality of life, inhibit pro-inflammatory factors, and enhance the expressions of 5-HT and BDNF, with no adverse event.

RESUMEN

OBJECTIVE@#To explore the influencing factors and countermeasures of infection in leukemia patients after allogeneic peripheral blood hematopoietic stem cell transplantation.@*METHODS@#A total of 126 patients with leukemia admitted in our hospital from August 2016 to March 2018 were selected. The number of infected patients after transplantation was recorded, and the causes of infection were analyzed.@*RESULTS@#Among the 126 patients, 43 were positive for infection, and the infection rate was 34.13%. A total of 89 pathogens were detected, of which bacteria accounted for 64.05%; virus accounted for 22.47%, and fungi accounted for 13.48%. The patient's age, donor type, pre-transplant infection, prophylactic use of antibiotics and aGVHD all were factors influencing the patient's infection (P＜0.05). The follow-up results showed that the incidence of infection in the intervention group significantly decreased after intervention with prevention program (P＜0.05). After reasonable nursing intervention, the incidence of infection in the intervention group after follow-up for 12 months was lower than that in the control group (P＜0.05).@*CONCLUSION@#Pre-transplant infection and prophylactic use of antibiotics are factors influencing the infection after allogeneic hematopoietic stem cell transplantation. The incidence of infection can be reduced by reasonable infection prevention.

Asunto(s)

Humanos , Enfermedad Injerto contra Huésped , Trasplante de Células Madre Hematopoyéticas , Infecciones , Leucemia , Trasplante de Células Madre de Sangre Periférica

RESUMEN

Objective To observe the effects of acupuncture at "Sanyinjiao" (SP6) and "Baihui" (GV20) on behaviors and contents of monoamine neurotransmitter dopamine (DA), noradrenalin (NE) and 5-hydroxytrypamine (5-HT) in prefrontal cortex (PFC) of attention deficit hyperactivity disorder (ADHD) rats; To discuss mechanism of action of acupuncture for prevention and treatment of ADHD. Methods 4-week-old SHR rats were randomly divided into the model group, Western medicine group and acupuncture group, and WKY rats with the same age were selected as the normal control group, with 10 rats in each group. Acupuncture was applied to "Sanyinjiao" (SP6) and "Baihui" (GV20) of rats in acupuncture group for 15 min. The rats in Western medicine group was given MPH (ritalin) for gavage, and rats were treated for 4 weeks in the dark-phase. Open field test (OFT), elevated plus maze (EPM) and novel object recognition test (NORT) were conducted to evaluate the spontaneous activity, impulsivity and learning-memorial ability of rats individually at the end of treatment. Contents of DA, NE and 5-HT in PFC were detected by HPLC. Results Compared with the normal control group, all of the behavior parameters of rats in model group were significantly higher (P＜0.05), and the contents of DA, NE and 5-HT in PFC of model group were lower than those of normal control group (P＜0.05). Compared with the model group, acupuncture group showed shorter movement distance, less rearing and fewer grooming activities in the OFT; percentages of times of entering the open arms and staying duration were reduced in the total period in the EPM; preference index in the NORT was elevated (P＜0.05); the contents of NE and 5-HT in PFC of acupuncture group increased significantly (P＜0.05). Conclusion Acupuncture at "Sanyinjiao" (SP6) and "Baihui" (GV20) canreduce the spontaneous activity and impulsivity, and improve learning and memory ability of ADHD model rats, which may be related to elevated contents of NE and 5-HT in the PFC.

RESUMEN

An analytical method was developed for determination of ginkgolic acids in Yinxing Tongzhi Dropping Pills by ultra high performance liquid chromatography-triple quadrupole mass spectrometry. The samples were purified by mix-mode anion exchange and reversed-phase SPE. A chromatographic column, Waters Cortecs T3 (50 mm×2.1 mm, 2.7 μm), was used with acetonitrile-methanol-1% acetic acid (44:44:12) as the mobile phase. The ginkgolic acids were detected by electrospray ionization mass spectrometry in negative mode with multiple reaction monitoring (MRM) mode. Ginkgolic acid C13:0, C15:1 and C17:1 possessed good linear correlation in the mass concentration range from 0.2 to 200 μg·L-1, 2 to 200 μg·L-1, 4 to 200 μg·L-1, respectively, with the correlation coefficients more than 0.999. The mean recoveries at spiked levels of 50, 250 and 600 μg·kg-1 were in the range of 70.8%-95.1%, and the RSDs were 0.7%-8.6%. The limits of quantification were 1, 10, 20 μg·kg-1, respectively. The method could be applied to the analysis of ginkgolic acids in complex matrix samples.

RESUMEN

OBJECTIVES: The aims of this study were to determine the effects of ω-3 fatty acids (ω-3FAs) on the Toll-like receptor 4 (TLR4)/nuclear factor κB p56 (NF-κBp56) signaling pathway in the pancreas of rats with severe acute pancreatitis (SAP). METHODS: Sixty-four Sprague-Dawley rats were randomly divided into 4 groups: the control, SAP-saline, SAP-soybean oil, and SAP-ω-FA groups. Severe acute pancreatitis was induced by the retrograde infusion of sodium taurocholate into the pancreatic duct. The expression of TLR4 and NF-κBp56 in the pancreas was evaluated by immunohistochemistry and Western blot analysis. The levels of the proinflammatory cytokines interleukin 6 and tumor necrosis factor α in the pancreas were measured by enzyme-linked immunosorbent assays. RESULTS: Toll-like receptor 4, NF-κBp56, and inflammatory cytokine expression in the pancreas was increased significantly in the SAP group compared with that in the control group (P < 0.05), but was significantly decreased in the ω-3FA group compared with that in the soybean oil group at 24 and 48 hours (P < 0.05). CONCLUSIONS: Our results suggest that during the initial stage of SAP ω-3FAs could efficiently lower the inflammatory response by activating the TLR4/NF-κBp56 signaling pathway.

Asunto(s)

Ácidos Grasos Omega-3/farmacología , FN-kappa B/metabolismo , Pancreatitis/metabolismo , Transducción de Señal/efectos de los fármacos , Receptor Toll-Like 4/metabolismo , Animales , Western Blotting , Citocinas/metabolismo , Ácidos Grasos Omega-3/administración & dosificación , Inmunohistoquímica , Mediadores de Inflamación/metabolismo , Masculino , Distribución Aleatoria , Ratas Sprague-Dawley , Índice de Severidad de la Enfermedad

RESUMEN

Drugs play the pharmacological effects by combining with target proteins. Identification of drug-target interactions is important for discovering new functions of drugs. In this paper, the target fingerprints based on molecular substructure and the drug-target similarity based on fingerprints are proposed to a random forest-based classification method, in order to identify the drug-target interactions. Experiments on enzymes, ion channels, G protein-coupled receptors and nuclear receptors proved the effectiveness of the proposed method. In addition, the proposed method is applied to predict the interactions between ingredients and targets of traditional Chinese medicines.

RESUMEN

AIM: To determine the effects of ω-3 fatty acids (ω-3FA) on the toll-like receptor 4 (TLR4)/nuclear factor κB p56 (NF-κBp56) signal pathway in the lungs of rats with severe acute pancreatitis (SAP). METHODS: A total of 56 Sprague-Dawley rats were randomly divided into 4 groups: control group, SAP-saline group, SAP-soybean oil group and SAP-ω-3FA group. SAP was induced by the retrograde infusion of sodium taurocholate into the pancreatic duct. The expression of TLR4 and NF-κBp56 in the lungs was evaluated by immunohistochemistry and Western blot analysis. The levels of inflammatory cytokines interleukin-6 and tumor necrosis factor-alpha in the lungs were measured by enzyme-linked immunosorbent assay. RESULTS: The expression of TLR4 and NF-κBp56 in lungs and of inflammatory cytokines in serum significantly increased in the SAP group compared with the control group (P < 0.05), but was significantly decreased in the ω-3FA group compared with the soybean oil group at 12 and 24 h (P < 0.05). CONCLUSION: During the initial stage of SAP, ω-3FA can efficiently lower the inflammatory response and reduce lung injury by triggering the TLR4/NF-κBp56 signal pathway.

Asunto(s)

Lesión Pulmonar Aguda/prevención & control , Aceites de Pescado/farmacología , Pulmón/efectos de los fármacos , FN-kappa B/metabolismo , Pancreatitis/tratamiento farmacológico , Receptor Toll-Like 4/metabolismo , Enfermedad Aguda , Lesión Pulmonar Aguda/etiología , Lesión Pulmonar Aguda/metabolismo , Lesión Pulmonar Aguda/patología , Amilasas/sangre , Animales , Modelos Animales de Enfermedad , Mediadores de Inflamación/metabolismo , Interleucina-6/metabolismo , Pulmón/metabolismo , Pulmón/patología , Masculino , Pancreatitis/inducido químicamente , Pancreatitis/metabolismo , Ratas Sprague-Dawley , Índice de Severidad de la Enfermedad , Ácido Taurocólico , Triglicéridos , Factor de Necrosis Tumoral alfa/metabolismo

RESUMEN

AIM: To assess the effect of inhibition of caspase-1 on acute renal injury in rats with severe acute pancreatitis (SAP). METHODS: Forty-two Sprague-Dawley rats were randomly divided into three groups: healthy controls (HC, n = 6), SAP rats treated with saline (SAP-S, n = 18), or SAP rats treated with a caspase-1/interleukin (IL)-1ß-converting-enzyme (ICE) inhibitor (SAP-I-ICE, n = 18). SAP was induced by retrograde infusion of 5% sodium taurocholate into the bile-pancreatic duct. HC rats were subjected to identical treatment and surgical procedures without sodium taurocholate. Rats received an intraperitoneal injection of isotonic saline (SAP-S) or the inhibitor (SAP-ICE-I) at 2 and 12 h after induction of acute pancreatitis. Surviving rats were sacrificed at different time points after SAP induction; all samples were obtained and stored for subsequent analyses. The levels of blood urea nitrogen (BUN) and creatinine (Cr) were measured using automatic methods, and serum IL-1ß concentrations were measured by an enzyme-linked immunosorbent assay. Intrarenal expression of IL-1ß, IL-18 and caspase-1 mRNAs was detected by RT-PCR. IL-1ß protein expression and the pathologic changes in kidney tissues were observed by microscopy after immunohistochemical or hematoxylin and eosin staining, respectively. RESULTS: The serum levels of BUN and Cr in the SAP-S group were 12.48 ± 2.30 mmol/L and 82.83 ± 13.89 µmol/L at 6 h, 23.53 ± 2.58 mmol/L and 123.67 ± 17.67 µmol/L at 12 h, and 23.60 ± 3.33 mmol/L and 125.33 ± 21.09 µmol/L at 18 h, respectively. All were significantly increased compared to HC rats (P < 0.01 for all). Levels in SAP-ICE-I rats were significantly decreased compared to SAP-S rats both at 12 and 18 h (P < 0.01 for all). Serum IL-1ß levels in the SAP-S group were 276.77 ± 44.92 pg/mL at 6 h, 308.99 ± 34.95 pg/mL at 12 h, and 311.60 ± 46.51 pg/mL at 18 h; all significantly higher than those in the HC and SAP-ICE-I groups (P < 0.01 for all). Intrarenal expression of IL-1ß mRNA was weak in HC rats, but increased significantly in SAP-S rats (P < 0.01). ICE inhibition significantly decreased the expression of IL-1ß and IL-18 mRNAs (P < 0.05 for all vs SAP-S), whereas caspase-1 mRNA expression was not significantly different. Weak IL-1ß immunostaining was observed in HC animals, and marked staining was found in the SAP-S group mainly in renal tubular epithelial cells. IL-1ß immunostaining was significantly descended in SAP-ICE-I rats compared to SAP-S rats (P < 0.05). Caspase-1 inhibition had no effect on the severity of kidney tissue destruction. CONCLUSION: The expression of caspase-1-activated cytokines IL-1ß and IL-18 plays a pivotal role in acute renal injury in rats with experimental SAP. Caspase-1 inhibition improves renal function effectively.

Asunto(s)

Lesión Renal Aguda/prevención & control , Caspasa 1/metabolismo , Inhibidores de Caspasas/farmacología , Riñón/efectos de los fármacos , Pancreatitis/tratamiento farmacológico , Enfermedad Aguda , Lesión Renal Aguda/sangre , Lesión Renal Aguda/enzimología , Lesión Renal Aguda/etiología , Lesión Renal Aguda/fisiopatología , Animales , Nitrógeno de la Urea Sanguínea , Creatinina/sangre , Citoprotección , Modelos Animales de Enfermedad , Interleucina-18/genética , Interleucina-18/metabolismo , Interleucina-1beta/sangre , Interleucina-1beta/genética , Riñón/enzimología , Riñón/patología , Riñón/fisiopatología , Masculino , Pancreatitis/sangre , Pancreatitis/inducido químicamente , Pancreatitis/enzimología , ARN Mensajero/metabolismo , Ratas Sprague-Dawley , Serpinas/farmacología , Índice de Severidad de la Enfermedad , Ácido Taurocólico , Proteínas Virales/farmacología

RESUMEN

Oculocerebrorenal syndrome of Lowe (OCRL) is an X-linked recessive disorder. This study investigated the history of a Chinese family with OCRL and used direct DNA sequencing to screen all exons of OCRL gene for mutations. A missense mutation (1736 A→G) in exon 15 was revealed, which resulted in the change of His (H) 507 to Arg (R). The patient's mother was the carrier of the heterozygous mutation in X-chromosome. To our knowledge, H507R mutation in OCRL gene has not been reported in Chinese people.

Asunto(s)

Humanos , Lactante , Masculino , Análisis Mutacional de ADN , Mutación Missense , Síndrome Oculocerebrorrenal , Genética , Monoéster Fosfórico Hidrolasas , Genética

RESUMEN

With the development of bio-technological drugs, drug immunogenicity evaluation has become key factor of clarifying safety and efficacy of these drugs. It has become the focus to establish a stable and reliable evaluation system. Due to the advantages such as continuous real-time monitoring, surface plasmon resonance (SPR) technology has been widely used in bio-technological drugs immunogenicity assessments. Our study applied this technology to detect anti-drug antibody (ADA) of a recombinant human anti-rabies monoclonal antibody NM57 in the sera of 48 volunteers admitted in phase I clinical trials. This method could satisfy the basic requirements of detection of ADA.

Asunto(s)

Humanos , Anticuerpos Antiidiotipos , Sangre , Alergia e Inmunología , Anticuerpos Monoclonales , Sangre , Alergia e Inmunología , Anticuerpos Antivirales , Sangre , Alergia e Inmunología , Virus de la Rabia , Alergia e Inmunología , Proteínas Recombinantes , Sangre , Alergia e Inmunología , Resonancia por Plasmón de Superficie

RESUMEN

Chromatin immunoprecipitation (ChIP) is an effective technique to analyze the interactions of DNA binding proteins with the genome in vivo. ChIP coupled with high density microarray (ChIP-chip) or high-throughput sequencing (ChIP-Seq) has generated large amount of data and expected to allow the development of a network describing the cellular transcriptional regulation. Here, we reviewed the ChIP, ChIP-chip, and ChIP-Seq techniques as well as their perspectives. Focus is given to data analysis of ChIP-Seq and the applications of ChIP-chip and ChIP-Seq.

Asunto(s)

Inmunoprecipitación de Cromatina , ADN/metabolismo , Proteínas/metabolismo , Animales , Humanos , Unión Proteica

RESUMEN

Objective To investigate the effects of arsenic trioxide(As2O3)combined with ascorbic acid(AA) on the apoptosis and differentiation of myelocytic leukaemia cells.Methods The acute promyelecytic leukaemia cell lines (NB4 and MR2)and erythroleukemia cell line(KS62)were cultured in vitro.Grouping wasbased on different concentration of As2O3(0,0.1,0.2,0.5,1.0 μmol/L),which WaS used a8 control groups.Then,AA(113.0μmol/)was added into each group.Cell morphologic changes of cell lines NB4,MR2 and K562 were observed under light microscope;The apoptosis symbols [Annexin V(+)/PI(-),Annexin V(+)/PI(+)]and differentiation symbols(CD11b and CD33)were detected by flow cytometry 96 hours later.Results

RESUMEN

To develop an EMSA-free assay approach for analyzing the sequence-specific DNA-binding proteins (DBPs), an easy cost-effective dsDNA-coupled plate (dcPlate) was developed in our lab for this purpose. In this paper, the assay conditions of such dcPlate were fully optimized for detecting an important transcription factor, NF-kappaB. The optimized parameters of dcPlate for assay of NF-kappaB were as follows: immobilized DNA probe at the concentration of 25 pmol/100 microL-well, incubation time of 90 min for NF-kappaB binding to dcPlate, primary and secondary antibody concentration of 0.1 microL/100 microL dilution, incubation time of 90 min for primary antibody binding to NF-kappaB, temperature of 25 degrees C for the above process, colorimetric developing time for 30 min. After optimization, the signal was improved three times higher than that from not optimized conditions. The linear colorimetric detection ranges of the purified recombinant NF-kappaB p50 and the cell nuclear extract were from 0.59 to 75 ng/well and 0.313 to 10 microg/well, respectively.

Asunto(s)

ADN/química , Subunidad p50 de NF-kappa B/análisis , Análisis de Secuencia por Matrices de Oligonucleótidos/instrumentación , Análisis de Secuencia por Matrices de Oligonucleótidos/métodos , Colorimetría , Humanos , Proteínas Recombinantes/análisis , Propiedades de Superficie

RESUMEN

<p><b>OBJECTIVE</b>To explore the molecular mechanism and prevention of retinoic acid syndrome (RAS).</p><p><b>METHODS</b>SDF-1 alpha mRNA from healthy adult lung tissue was measured by RT-PCR, CXCR4 protein expression on the cell membrane of APL cells induced by ATRA (APL-ATRA) was tested by FCM, and the rotary cell culture system (RCCS) was used to build a modal for in vitro stimulation of APL-ATRA infiltrating human lung tissue. The ability of APL-ATRA in adhesion, migration and infiltration was observed by interference from DEX, Ara-C and DNR.</p><p><b>RESULTS</b>The APL-ATRA cells could evidently infiltrate into normal lung tissue. Mean fluorescence intensity (MFI) of CXCR4 on the cell membrane of APL-ATRA cells was 30.6 +/- 1.8, which was much higher than that on unspecialized APL cells (9.8 +/- 4.2). SDF-1 alpha mRNA expression was detected positive in all 6 lung tissue. Contrary to the control groups, DEX could dramatically restrain the ability of APL-ATRA cells in adhesion and migration [(27.2 +/- 2.6)% vs. (46.0 +/- 3.0)%, (28.1 +/- 4.0)% vs. (48.2 +/- 3.0)%], while Ara-C and DNR could distinctly depress the ability in adhesion, migration and infiltration [(28.1 +/- 3.0)%, (30.2 +/- 3.2)% vs. (46.0 +/- 3.0)%; (29.0 +/- 4.0)%, (23.0 +/- 5.2)% vs. (48.2 +/- 3.0)%; (16.8 +/- 7.6)%, (17.1 +/- 6.0)% vs. (43.6 +/- 5.0)%].</p><p><b>CONCLUSION</b>In vitro APL-ATRA cells can infiltrate into the human lung tissue. High expression of CXCR4 on APL-ATRA and SDF-1 alpha in the lung tissue may be one of the molecular mechanisms of the lung infiltration and RAS. DEX, Ara-C and DNR can dramatically restrain the ability of APL-ATRA cells in adhesion, migration and infiltration.</p>

Asunto(s)

Adolescente , Adulto , Niño , Femenino , Humanos , Masculino , Persona de Mediana Edad , Adhesión Celular , Técnicas de Cultivo de Célula , Movimiento Celular , Quimiocina CXCL12 , Genética , Metabolismo , Leucemia Promielocítica Aguda , Metabolismo , Patología , Invasividad Neoplásica , Receptores CXCR4 , Genética , Metabolismo , Tretinoina , Células Tumorales Cultivadas

RESUMEN

The mainly antigenic sites for the adenovirus neutraliation are present on Loop1 and Loop2 of hexon.Majority research were focus in the human adenovirus.Little was known on infectious canine hepatitis virus (ICHV), which was also called canine adenovirus typeⅠ.Here,ICHV (the isolated strain) DNA was isolated and purified from the cultured MDCK cells.The Loop1 and Loop2 fragments were amplified by polymerase chain reaction(PCR) method,and then was connected by ligase T4.The target fragment was then connected with vector pET28a.The nucleotide sequence ecoding Loop1 and Loop2 was determined.The nucleotide sequence identity of Loop1 region between the isolated strain and CLL, RI261 and Toronto A26/61 strains is 100%, 100% and 83.8%, and the nucleotide sequence identity of Loop2 region between the isolated strain and CLL, RI261 and Toronto A26/61 strains is 88.1% , 88.1% and 99.3%, and amino acid identity is 93.6%, 93.6% and 98.6%.The recombinant Loop protein was expressed in E.coli and was approximately 36kDa in size,and then was purified. Then BALB/c mice were injected subcutaneously in the back and armpit with the recombinant Loop protein.The anti-ICHV antibody titers of immunized serum was tested by indirect ELISA and the titers were up to 1:320.Western blot demonstrated that immunized sera could specifically combine with ICHV. The research laid a foundation for creating new genetic engineering products of infectious canine hepatitis virus.

RESUMEN

Comparing the fluorescence spectra of tobacco peroxidase I (TOP I) solution and the solutions titrated by iodine anions, the number of binding locus and the binding constant of iodine anion to TOP I were calculated by using the modified Stern-Volmer equation. The mechanism of the quenching of fluorescence spectra and the distribution of tryptophane residues in the TOP I molecule were discussed.

Asunto(s)

Nicotiana/enzimología , Peroxidasa/química , Proteínas de Plantas/química , Espectrometría de Fluorescencia/métodos , Algoritmos , Aniones/química , Concentración de Iones de Hidrógeno , Yodo/química , Cinética , Peroxidasa/metabolismo , Proteínas de Plantas/metabolismo , Unión Proteica , Proyectos de Investigación , Temperatura

RESUMEN

The ultraviolet/visible spectra of TOP I were studied. It was proved that TOP I is an acid enzyme containing it hemochrome as an agon. When the pH reduced, the Soret absorption in UV-Vis region exhibited a blue shift, when pH increased exhibited a red shift. The result of the influence of carbamide, the denaturant, on the spectra showed that TOP I may have a unfolded structural change in the solution, which made the peptide chain completely extend. After adding Fe(III), Fe(II), Cu(II), Zn(II), Co(II), Ni(II) and Sn(II) to the apo-TOP I, the UV-Vis spectra changed except for Fe(III), which almost did not change at all. The strongest characteristic absorption peak of the Soret showed a blue shift to different extent and it became weak gradually. The situation of alpha-strip and beta-strip remained unchanged while the relative strength decreased.

Asunto(s)

Nicotiana/enzimología , Peroxidasas/química , Espectrofotometría Ultravioleta/métodos , Dicroismo Circular/métodos , Modelos Moleculares , Estructura Molecular , Unión Proteica/fisiología , Conformación Proteica , Espectrofotometría Infrarroja/métodos , Espectrometría Raman

RESUMEN

Tobacco peroxidase (TOPI) has been purified by ammonium sulfate fractionation and column chromatography, involving ion exchange on DE-52 cellulose, gelfiltration on Sephadex G-75 and ion exchange on DEAE-sephadex A-50. The specific activity of peroxidase purified was 4,826 U/mg. It has been demonstrated by SDS-PAGE as a single band. Its molecular weight (matrix assisted laser desorption/ionization time of flight mass spectra) and isoelectric point (pI) have been determined to be 21,888.5 and 3.5 respectively. The native enzyme is one of a haemachrome-containing acidic protein and has its soret maximum at 402 nm and alpha and beta bands at 636 nm and 498 nm, respectively. Its characteristic absorption spectra and fluorescence spectra changed in different pH and denaturant. It has its own characteristic absorption spectra and fluorescence spectra.

Asunto(s)

Nicotiana/enzimología , Peroxidasa/química , Punto Isoeléctrico , Peroxidasa/aislamiento & purificación , Espectrometría de Fluorescencia , Espectrofotometría Ultravioleta