RESUMO
OBJECTIVE: To evaluate the efficacy and safety of using Jiangzhi Tongluo Soft Capsule (JTSC) combined with Atorvastatin Calcium Tablet (ACT) or ACT alone in treatment of combined hyperlipidemia. METHODS: A randomized, double blinded, parallel control, and multi-center clinical research design was adopted. Totally 138 combined hyperlipidemia patients were randomly assigned to the combined treatment group (A) and the atorvastatin treatment group (B) by random digit table, 69 in each group. All patients took ACT 20 mg per day. Patients in the A group took JTSC 100 mg each time, 3 times per day. Those in the B group took JTSC simulated agent, 100 mg each time, 3 times per day. The treatment period for all was 8 weeks. Serum levels of triglyceride (TG), total cholesterol (TC), low density lipoprotein cholesterol (LDL-C), and high density lipoprotein cholesterol (HDL-C) were observed before treatment, at week 4 and 8 after treatment; and safety was assessed as well. RESULTS: At week 4 and 8 after treatment serum TG decreased by 26.69% and 33.29% respectively in the A group (both P < 0.01), while it was decreased by 25.7% and 22.98% respectively in the B group (both P < 0.01). At week 8 decreased serum TG was obviously higher in the A group than in the B group (P < 0.05). Compared with before treatment, serum levels of LDL-C and TC levels decreased significantly in the two groups (all P < 0.01). There was no statistical difference in the drop-out value and the drop-out rate of serum LDL-C and TC levels (P > 0.05). At week 8 the serum HDL-C level showed an increasing tendency in the two groups. No obvious increase in peptase or creatase occurred in the two groups after treatment. CONCLUSION: JTSC combined with ACT could lower the serum TG level of combined hyperlipidemia patients with safety.
Assuntos
Medicamentos de Ervas Chinesas/uso terapêutico , Ácidos Heptanoicos/uso terapêutico , Hiperlipidemias/tratamento farmacológico , Pirróis/uso terapêutico , Adulto , Atorvastatina , Método Duplo-Cego , Quimioterapia Combinada , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Resultado do Tratamento , Triglicerídeos/sangueRESUMO
AIM: To transform the human anti-rabies virus glycoprotein (anti-RABVG) single-chain variable fragment (scFv) into a Fab fragment and to analyze its immunological activity. METHODS: The Fab gene was amplified using overlap PCR and inserted into the vector pComb3XSS. The recombinant vector was then transformed into E coli Top10F' for expression and purification. The purified Fab was characterized using SDS-PAGE, Western blotting, indirect ELISA, competitive ELISA, and the fluorescent antibody virus neutralization test (FAVN), respectively, and examined in a Kunming mouse challenge model in vivo. RESULTS: A recombinant vector was constructed. The Fab was expressed in soluble form in E coli Top10F'. Specific binding of the Fab to rabies virus was confirmed by indirect ELISA and immunoprecipitation (IP). The neutralizing antibody titer of Fab was 10.26 IU/mL. The mouse group treated with both vaccine and human rabies immunoglobulin (HRIG)/Fab091 (32 IU/kg) showed protection against rabies, compared with the control group (P<0.05, Logrank test). CONCLUSION: The antibody fragment Fab was shown to be a neutralizing antibody against RABVG. It can be used together with other monoclonal antibodies for post-exposure prophylaxis of rabies virus in future studies.
Assuntos
Anticorpos Neutralizantes/imunologia , Anticorpos Antivirais/imunologia , Fragmentos Fab das Imunoglobulinas/imunologia , Vacina Antirrábica/administração & dosagem , Vírus da Raiva/imunologia , Raiva/prevenção & controle , Animais , Anticorpos Neutralizantes/administração & dosagem , Anticorpos Neutralizantes/genética , Anticorpos Antivirais/administração & dosagem , Anticorpos Antivirais/genética , Humanos , Fragmentos Fab das Imunoglobulinas/administração & dosagem , Fragmentos Fab das Imunoglobulinas/genética , Camundongos , Vírus da Raiva/genéticaRESUMO
We report a simple electrochemical method for the identification of properties of peripheral nerve fibers, based on the detection of a neurotransmitter enzyme, acetylcholinesterase (AChE). A poly(diallydimethylammonium) (PDDA) adulterated poly(dimethylsiloxane) (PDMS) film is spin-coated on the surface of gold electrodes. Gold nanoparticles (AuNPs) are in situ synthesized on the polymer film, which act as "electron antennae" between the film and the electrode surface and also provide a biocompatible interface. This PDMS-PDDA/AuNPs film shows different adsorption sites to choline oxidase (ChO) and AChE; after incubation with ChO, the polymer-gold nanocomposite film also shows excellent adsorption ability to AChE. Moreover the adsorption sites of AChE would not be blocked by bovine serum albumin (BSA) which provides a good platform for the quantitative amperometric determination of AChE via the oxidation of the enzymatically generated H 2O 2 in the bienzyme system in the presence of acetylcholine. The detection limit is down to 1.0 unit/mL. The polymer-gold nanocomposite film shows excellent anti-interference ability to the coexistent electroactive substances such as ascorbic acid. Thus it was applied to determine AChE in peripheral nerve fibers homogenates and identify the motor and sensory fibers for the first time. Compared with histochemical staining methods, the electrochemical technique shows good accurate rate and faster response, which has good potential for a clinical application.
Assuntos
Materiais Biocompatíveis , Eletroquímica/métodos , Ouro/química , Nanopartículas Metálicas , Fibras Nervosas/química , Nervos Periféricos/química , Polímeros/química , Animais , Cães , Microscopia Eletrônica de Varredura , Sensibilidade e EspecificidadeRESUMO
Removal of arsenate from water using granular ferric hydroxide (GFH) was investigated under different pH and As(V) loading conditions, using batch equilibrium adsorption, FTIR, and EXAFS methods. The arsenate adsorption envelopes on GFH exhibited broad adsorption maxima when the initial As(V) concentration was less than 500 mg/L at sorbent concentration of 10 g/L. As the initial As(V) concentration increased to 500, 1000 or 2000 mg/L for the same sorbent concentration, distinct adsorption maxima appeared and shifted to lower pH. Acidimetric-alkalimetric titration and arsenic adsorption isotherm data indicated that the surface of GFH is high heterogeneous. FTIR spectra revealed that complexes of two different structures, bidentate and monodentate, were formed upon the adsorption of arsenate on GFH, and bidentate complexes were only observed at pH values greater than 6. The EXAFS analyses confirmed that arsenate form bidentate binuclear complexes with GFH at pH 7.4 as evidenced by an average Fe-As(V) bond distance of 3.32 A.
Assuntos
Arsênio/isolamento & purificação , Compostos Férricos/química , Poluentes Químicos da Água/isolamento & purificação , Adsorção , Microscopia Eletrônica de Varredura , Espectroscopia de Infravermelho com Transformada de FourierRESUMO
The title compound, [MnCl(2)(C(12)H(8)N(2)O(2))(2)], displays a novel supramolecular chain formed by intermolecular O-H...Cl hydrogen bonds and aromatic stacking. The molecule has crystallographically imposed twofold symmetry with the Mn(II) atom on the twofold axis. In the 1,10-phenanthroline-5,6-diol ligand, each H atom of the two hydroxy groups is oriented towards the other hydroxy O atom. Both hydroxy groups form intermolecular O-H...Cl hydrogen bonds with a single Cl atom of an adjacent molecule. These hydrogen bonds connect the molecules via operation of the molecular twofold axis and the centre of inversion of the crystal lattice, forming a doubly-bridged one-dimensional structure with Mn atoms as the nodes. Strong aromatic pi-stacking between two antiparallel neighbouring 1,10-phenanthroline-5,6-diol ligands also helps to stabilize the chain.
RESUMO
Sodium pyrophosphate (pyro-P, Na4P2O7), sodium tripolyphosphate (tripoly-P, Na5P3O10), and sodium hexametaphosphate (meta-P, (NaPO3)6) were selected as the model compounds of condensed phosphate to investigate the adsorption behavior of condensed phosphate on aluminum hydroxide. The adsorption was found to be endothermic and divisible into two stages: (1) fast adsorption within 1 h; and (2) slow adsorption between 1 and 24 h. The modified Freundlich model simulated the fast adsorption stage well; the slow adsorption stage was described well by the first-order kinetics. The activation energies of pyro-P, tripoly-P, and meta-P adsorption on aluminum hydroxide were determined to be 20.2, 22.8 and 10.9 kJ/mol P adsorbed, respectively, in the fast adsorption stage and to be 66.3, 53.5 and 72.5 kJ/mol P adsorbed, respectively, in the slow adsorption stage. The adsorption increased the negative charge of the aluminum hydroxide surface. Transmission electron microscopy and energy dispersive X-ray analysis analyses provided evidence that the adsorption was not uniform on the surface and that the small crystals contributed more to the fast adsorption than the normal sites did. The results from X-ray fluorescence spectrometry and X-ray photoelectron spectroscopy tests also revealed the uneven adsorption of condensed phosphate as a function of the penetration depth. More condensed phosphates were adsorbed on the outer surface of aluminum hydroxide than in its inner parts.
Assuntos
Hidróxido de Alumínio/química , Fosfatos/química , Poluentes Químicos da Água/química , Adsorção , Concentração de Íons de Hidrogênio , Cinética , TemperaturaRESUMO
Information on the binding of organic ligands to metal (hydr)oxide surfaces is useful for understanding the adsorption behaviour of natural organic matter on metal (hydr)oxide. In this study, benzoate and salicylate were employed as the model organic ligands and aluminum hydroxide as the metal hydroxide. The attenuated total reflectance Fourier transform infrared (ATR-FTIR) spectra revealed that the ligands benzoate and salicylate do coordinate directly with the surface of hydrous aluminum hydroxide, thereby forming inner-sphere surface complexes. It is concluded that when the initial pH is acidic or neutral, monodentate and bridging complexes are to be formed between benzoate and aluminum hydroxide while bridging complexes predominate when the initial pH is alkalic. Monodentate and bridging complexes can be formed at pH 5 while precipitate and bridging complexes are formed at pH 7 when salicylate anions are adsorbed on aluminum hydroxide. The X-ray photoelectron (XP) spectra demonstrated the variation of C 1s binding energy in the salicyate and phenolic groups before and after adsorption. It implied that the benzoate ligands are adsorbed through the complexation between carboxylate moieties and the aluminum hydroxide surface, while both carboxylate group and phenolic group are involved in the complexation reaction when salicylate is adsorbed onto aluminum hydroxide. The information offered by the XPS confirmed the findings obtained with ATR-FTIR.
Assuntos
Hidróxido de Alumínio/química , Benzoato de Sódio/química , Salicilato de Sódio/química , Adsorção , Espectroscopia de Infravermelho com Transformada de FourierRESUMO
The effects of orthophosphate on the adsorption of natural organic matter (NOM) on aluminum hydroxide were investigated using three organic compounds as surrogates, including humic acid (HA), phthalic acid, and 2,3-dihydroxybenzoic acid (2,3-DHBA). The adsorption of phthalic acid and 2,3-DHBA was very limited compared to that of HA, whereas their adsorption was reduced much more significantly than that of HA by phosphate. The efficiency of phosphate in reducing HA adsorption increased with increasing phosphate concentration. Phosphate adsorption was slightly reduced by phthalic acid and 2,3-DHBA but moderately suppressed by HA. The adjacent carboxylic groups mainly contributed to the adsorption of humic acid at low pH, while the adjacent phenol groups were responsible for the adsorption of humic acid at high pH. HPLC-SEC and SUVA analysis revealed that HA molecules with high molecular weight were adsorbed preferentially but were easily displaced by the specifically adsorbed phosphate. TM-AFM images revealed that the aggregation of HA molecules and the protonation of carboxylic groups at low pH facilitated the adsorption under acidic conditions. The presence of phosphate increases the coagulant dosage for NOM removal as some sites on the coagulant precipitates become utilized by phosphate.
RESUMO
The adsorption of a series of aromatic carboxylic acids, with different numbers and positions of carboxyl groups in the phenyl ring, on aluminum hydroxide was investigated with ATR-FTIR and kinetic analyses to verify the role of aromatic carboxyl groups in the surface complexation of NOM with aluminum hydroxide. It was revealed that the formation of outer-sphere complexes dominated the adsorption of most of the aromatic carboxylates over the pH range examined in this study. Inner-sphere complexes were only detected at some pH levels for some aromatic carboxylates adsorption. The aromatic carboxylates were most likely to be adsorbed onto the first surface layer of hydroxyl groups and water molecules without forming coordinative bonds with the aluminum hydroxide surfaces, but strong hydrogen bonds were formed in this process. The adsorption data fitted the pseudo-second-order kinetic model very well. The activation energies of adsorption calculated from the rate constants of pseudo-second-order kinetics agreed with the ATR-FTIR analysis that the aromatic carboxylates, except pyromellitate, were adsorbed predominantly as outer-sphere complexes on aluminum hydroxide at pH 7. This study revealed that phenolic groups may be more significant than carboxylic groups for the chemical adsorption of NOM onto minerals. The presence of additional carboxylic groups enhanced the adsorption considerably, while the enhancement was limited when there were three or four carboxylic groups on the phenyl ring. The adsorption of aromatic carboxylates was affected by the positions and pK(a) values of the donor groups and the solution pH.
Assuntos
Hidróxido de Alumínio/química , Ácidos Carboxílicos/química , Compostos Orgânicos/química , Poluentes Químicos da Água/química , Adsorção , Concentração de Íons de Hidrogênio , Cinética , Estrutura Molecular , Espectroscopia de Infravermelho com Transformada de Fourier , Propriedades de SuperfícieRESUMO
The adsorption isotherm of and the pH effect on the adsorption of myo-inositol hexaphosphate (myo-IP6) on amorphous aluminum hydroxide was investigated. It was found that the adsorption isotherm of myo-IP6 on aluminum hydroxide could be well fitted with the Freundlich isotherm. The amount of myo-IP6 adsorbed remained almost constant in the range of pH 4.0 to 7.0, but it decreased considerably as the initial pH was over 7. The adsorption of myo-IP6 resulted in an increase in the pH level due to the release of OH(-) ions, which suggested that the adsorption of myo-IP6 on aluminum hydroxide was caused by a ligand exchange reaction. ATR-FTIR analysis of myo-IP6 in solution and adsorbed on aluminum hydroxide at different pH were performed. The ATR-FTIR investigation indicated that myo-IP6 was adsorbed onto aluminum hydroxide by forming inner-sphere complexes and adsorption facilitated the deprotonation of phosphate groups. The asymmetric vibration of the PO bond in AlPO(-)(3) appearing at a lower frequency than that in the terminal HPO(-)(3) indicated that Al bound to the O atom not as strongly as the H atom did. The ATR-FTIR investigation and theoretical calculation (with the Gaussian 03 program) revealed that three of the six phosphate groups in myo-IP6 molecules were bound to aluminum hydroxide while the other three remained free when myo-IP6 was adsorbed on aluminum hydroxide.
Assuntos
Hidróxido de Alumínio/química , Ácido Fítico/análogos & derivados , Ácido Fítico/química , Concentração de Íons de Hidrogênio , Modelos Moleculares , Conformação Molecular , Soluções , Espectroscopia de Infravermelho com Transformada de FourierRESUMO
The role of phenolic groups in the interaction of natural organic matter (NOM) with metal hydroxides was investigated with ATR-FTIR spectroscopy and adsorption tests by employing a series of dihydroxybenzoic acids (DHBAs) as the NOM surrogates and aluminum hydroxide as the adsorbent. All DHBAs examined in this study were found to be adsorbed on aluminum hydroxide by forming inner-sphere complexes. Carboxylic groups governed the complexation of DHBAs with aluminum hydroxide at low pH or in cases when the two hydroxyl groups were not adjacent to each other and neither of them was ortho to the carboxylic group. The involvement of the phenolic groups, ortho to another phenolic group or ortho to the carboxylic groups, in the complexation increased with increasing pH as the deprotonation of phenolic groups was easier at higher pH. The presence of phenolic groups increased the electron density of the carboxylic groups and facilitated the inner-sphere complexation of the carboxylic groups with metal hydroxide. The correlation between the pK(a) values and the amount of organic acid adsorbed on the aluminum hydroxide revealed that the adsorption of DHBAs at acidic pH was largely dependent on the surface chelate formation rather than on the electronic effect.
Assuntos
Hidróxido de Alumínio/química , Modelos Químicos , Espectroscopia de Infravermelho com Transformada de Fourier/métodos , Fenóis/químicaRESUMO
OBJECTIVE: To construct, express and purify human scFv antibody (S1) against the recombinant SAG1 of Toxoplasma gondii and fused to green fluorescent protein (GFP), and observe its binding capacity to tachyzoite of Toxoplasma gondii. METHODS: The GFP gene amplified from vector pEGFP-N1 was subcloned into procaryotic expression vector pET-26b (+), then the S1 scFv antibody gene amplified from phagmid pIT-2-S1 was cloned into downstream of GFP gene. The recombinant plasmid pET-26b-GFPS1 proved by DNA sequencing was transformed into E. coli BL21, and induced for fusion expression of GFPS1 with IPTG, the green fluorescence of E. coli BL21 harboring plasmid pET-26b-GFPS1 was observed under the fluorescence microscope. The expressed GFPS1 was purified with Ni2+ chelating HiTrap HP column, and detected with SDS-PAGE. Toxoplasma tachyzoites were incubated with the recombinant GFPS1, and the binding bioactivity was observed under the fluorescence microscope. RESULTS: The fused gene of S1 and GFP was successfully cloned into procaryotic expression vector pET-26b proved by DNA sequencing. The green fluorescence of E. coli BL21 harboring plasmid pET-26b-GFPS1 was catched under the fluorescence microscope. The recombinant GFPS1 protein about Mr 53 000 was expressed in E. coli as inclusion body. The immunofluorescence detection verified that anti-rSAG1 scFv antibody S1 could specifically bind outer membrane of Toxoplasma tachyzoite. CONCLUSIONS: The purified rGFPS1 shows a strong binding capacity to outer membrane of Toxoplasma tachyzoite using GFP as a labeling protein, and the constructed pET-26b-GFP can also be used for research on other targeting molecules.
Assuntos
Anticorpos Antiprotozoários/biossíntese , Antígenos de Protozoários/biossíntese , Proteínas de Fluorescência Verde/biossíntese , Proteínas de Protozoários/biossíntese , Toxoplasma/imunologia , Toxoplasma/metabolismo , Animais , Anticorpos Antiprotozoários/imunologia , Antígenos de Protozoários/imunologia , Expressão Gênica , Reação em Cadeia da Polimerase , Proteínas de Protozoários/imunologia , Proteínas Recombinantes de Fusão/biossíntese , Toxoplasma/genéticaRESUMO
To investigate the effect of pH on the oxidation of anilines by potassium permanganate, aniline and p-Chloroaniline were taken as the target contaminants, and the experiments were conducted under the condition with potassium permanganate in excess over a wide pH range. The reaction displayed remarkable autocatalysis, which was presumably ascribed to the formation of complexes by the in situ generated MnOx and the target contaminants on its surface, and thereby improved the oxidation rate of the target contaminants by permanganate. The reaction kinetics was fitted with the pseudo-first-order kinetics at different pH to obtain the pseudo-first-order reaction constants (k(obs)). The second-order rate constants calculated from permanganate concentration and k,b, increased with the increase of pH and reached the maximum near their respective pKa, after which they decreased gradually. This tendency is called parabola-like shaped pH-rate profile. The second-order rate constants between permanganate and anilines were well fitted by the proton transfer model proposed by us in previous work.
Assuntos
Compostos de Anilina/química , Compostos de Manganês/química , Óxidos/química , Permanganato de Potássio/química , Concentração de Íons de Hidrogênio , Cinética , OxirreduçãoRESUMO
This paper attempted to study the feasibility of reusing water treatment works sludge ("alum sludge") to improve particulate pollutant removal from sewage. The main issues focused upon were: (1) the appropriate dosage of the alum sludge, (2) the appropriate operating conditions, and (3) the possible mechanisms for enhancement by adding alum sludge. Actual alum sludge and sewage were applied to a series of jar tests conducted under various conditions. It has been found that both the SS and COD removal efficiencies could be improved by the addition of the alum sludge, which was mainly attributed to the removal of relatively fine particles with a size of 48-200 microm. The appropriate dosage of the alum sludge was determined to be 18-20 mg of Al/L. Increasing the mixing speed or reducing the floc size of the alum sludge enhanced the SS and COD removal and the dispersed alum sludge could remove particulate contaminants with smaller size than the raw sewage. ToF-SIMS evidence revealed that the aluminum species at the surface of the alum sludge were effectively utilized for improving the SS and COD removal. It was postulated that the sweep flocculation and/or the physical adsorption might play key roles in the enhancement of particulate pollutant removal from sewage.
Assuntos
Compostos de Alúmen , Esgotos , Eliminação de Resíduos Líquidos/métodos , Adsorção , Conservação dos Recursos Naturais , Floculação , Tamanho da PartículaRESUMO
Two model compounds, sodium pyrophosphate (pyro-P) and sodium tripolyphosphate (tripoly-P), were employed to elucidate the binding mechanisms of condensed phosphate on aluminum hydroxide by utilizing attenuated total reflectance-Fourier transform infrared (ATR-FTIR) spectroscopy. Peak assignments for the condensed phosphates in the solution phase and those adsorbed on the surface of aluminum hydroxide were made. Electron delocalization and polarization were employed to explain the peak shifts and the complexation of condensed phosphate with aluminum hydroxide. The tripoly-P and pyro-P were adsorbed on aluminum hydroxide by forming inner-sphere complexes. The adsorbed condensed phosphates were deprotonated in the pH range from 4 to 10. Monodentate, bidentate, and binuclear complexes were formed when pyro-P was adsorbed on aluminum hydroxide, while monodentate and binuclear complexes were formed when tripoly-P was adsorbed. Based on the FTIR data, we proposed that when either bidentate or binuclear complexes were formed, the two oxygen atoms participating in the complexation with aluminum hydroxide could not be originated from the same terminal phosphate moiety. The AlO bond formed in the complexation of pyro-P or tripoly-P with aluminum hydroxide (AlPO(-3)) was not as strong as the HO bond in terminal HPO(-3). The bridging PO(-2) of tripoly-P did not coordinate with aluminum hydroxide. The real-time ATR-FTIR study on condensed phosphate adsorption revealed that a long contact time between condensed phosphates and aluminum hydroxide particles can result in a transformation of an initially formed species into a thermodynamically more stable phase.
Assuntos
Hidróxido de Alumínio/química , Fosfatos/química , Adsorção , Sítios de Ligação , Polifosfatos/química , Sensibilidade e Especificidade , Soluções/química , Espectroscopia de Infravermelho com Transformada de Fourier/métodos , Propriedades de SuperfícieRESUMO
BACKGROUND: The development of new adjuvants for human use has been the focus of attention. This study's aim is to explore the possibility of using nanoparticle Ca nanoparticles (CA) as a vaccine adjuvant of anti-idiotypic antibody NP30 against schistosomiasis and its protective mechanisms. METHODS: Nanoparticle CA-NP30 conjugate (CA-NP30) was fabricated. BALB/c mice were immunized actively with CA-NP30 to evaluate its effects of protective immunity on mice. The serum levels of specific IgG, IgG1 and IgG2a antibodies against NP30 and the concentrations of IFN-gamma and IL-4 in supernatant of splenocytes were determined via ELISA. RESULTS: Nanoparticle CA could enhance significantly the protective immunity of NP30 against infection of Schistosoma japonicum and the worm reduction rose from 36.0% (NP30 alone) to 52.6%. The serum levels of specific IgG, IgG1 and IgG2a antibodies against NP30 increased remarkably, as compared with those of the group immunized with NP30 alone. The concentration of IFN-gamma in supernatant of splenocyte was drastically elevated [the groups immunized with CA-NP30 and NP30 alone were (493.80 +/- 400.74) pg/ml and (39.03 +/- 39.58) pg/ml, respectively], but the concentration of IL-4 showed no significant difference from that of NP30 alone [(27.94 +/- 9.84) pg/ml vs (27.28 +/- 14.44) pg/ml]. CONCLUSIONS: Nanoparticle CA could act as a vaccine adjuvant of anti-idiotypic antibody NP30 against schistosomiasis. The mechanism could be that CA-NP30 enhances humoral and cellular immune responses in mice.
Assuntos
Adjuvantes Imunológicos , Anticorpos Anti-Idiotípicos/imunologia , Nanotecnologia , Esquistossomose/prevenção & controle , Vacinas , Animais , Anticorpos Anti-Helmínticos/imunologia , Camundongos , Camundongos Endogâmicos BALB CRESUMO
OBJECTIVE: To construct single chain Fv (scFv) gene of anti-idiotypic monoclonal antibody NP30 of Schistosoma japonicum. METHODS: The heavy and light chain variable region genes of anti-idiotypic monoclonal antibody NP30 of Schistosoma japonicum were inserted into two corresponding sites of expression vector pTHA90, and a scFv gene was constructed with a short peptide (Gly4Ser)3 linker gene. The recombinants were determined by digesting with XhoI/SpeI, XbaI/EcoRI and XhoI/EcoRI, and then were introduced into E. coli Top10. The antigen binding activity of expressed product was detected with ELISA. RESULTS: The recombinants were determined by digesting with endonucleases and expected bands were identified. The value of expressed scFv was 3 times higher than negative control by ELISA(OD492 = 1.06). CONCLUSION: The scFv gene of anti-idiotypic monoclonal antibody NP30 of Schistosoma japonicum was successfully cloned, and the expressed scFv fragment could interact specifically with antigen NP48.
Assuntos
Anticorpos Anti-Idiotípicos/genética , Anticorpos Anti-Helmínticos/genética , Anticorpos Monoclonais/genética , Fragmentos de Imunoglobulinas/genética , Schistosoma japonicum/imunologia , Animais , Anticorpos Anti-Idiotípicos/biossíntese , Anticorpos Anti-Helmínticos/biossíntese , Anticorpos Monoclonais/biossíntese , Clonagem Molecular , Cadeias Pesadas de Imunoglobulinas/genética , Cadeias Leves de Imunoglobulina/genética , Região Variável de Imunoglobulina/genéticaRESUMO
A new coagulant was synthesized by oxidizing Fe(II) with H2O2 instantaneously. Key parameters in the preparation of the coagulant including aging time, acid volume, and n(H2O2 )/n(Fe) were investigated through jar tests. The optimized coagulant was then compared with Fe2 (SO4)3 and PFS (poly-ferric sulphate) in residual turbidity, UV254, Zeta potential to evaluate its coagulation efficiency and infer the reaction mechanism. Aging time has a little effect on coagulation efficiency. The optimized acid concentration was 0.1 mol x L(-1). Coagulation efficiency increased with n(H2O2)/n (Fe). 0.55 is the most economical and efficient molar ratio. The removal efficiency of turbidity of the newly-formed ferric is similar to Fe2 (SO4)3 or PFS. The removal efficiency of UV254 by newly-formed ferric is higher than that by Fe2 (SO4)3 by 20%-44%. The Zeta potential of supernatant after being treated by newly-formed ferric gradually increased to 0 mV with the increasing coagulant dosage, indicating a strong effect of charge neutralization. The newly-formed ferric is a promising new coagulant with an extremely short aging time, a simple synthesis process, a lower cost than Fe2 (SO4)3 or PFS by more than 50%, and a high removal efficiency of UV254 absorbance.
Assuntos
Água Potável , Compostos Férricos/química , Floculação/efeitos dos fármacos , Purificação da Água/métodos , Peróxido de Hidrogênio/químicaRESUMO
The murine monoclonal anti-idiotypic antibody NP30 is a promising therapeutic antibody against Schistosoma japonicum. However, the immunogenicity of murine NP30 limits its further study and application in humans. Here the chimeric Fab of NP30 (chFab-NP30) comprising the variable regions of murine NP30 and constant regions of human antibody was assembled. chFab-NP30 was expressed and purified as a soluble and functional protein. Administration of chFab-NP30 in vivo increased the survival rate, reduced egg burdens and ameliorated organ pathology of mice with acute schistosomiasis. Our study indicated that chFab-NP30 is a promising candidate to be used as a specific and efficient recombinant antibody against acute schistosomiasis japonica. Further studies on function mechanism of chFab-NP30 needs to be carried out in the future.
Assuntos
Anti-Helmínticos/administração & dosagem , Anticorpos Anti-Idiotípicos/administração & dosagem , Anticorpos Monoclonais/administração & dosagem , Schistosoma japonicum/efeitos dos fármacos , Esquistossomose Japônica/tratamento farmacológico , Animais , Anti-Helmínticos/isolamento & purificação , Anticorpos Anti-Idiotípicos/genética , Anticorpos Anti-Idiotípicos/isolamento & purificação , Anticorpos Monoclonais/genética , Anticorpos Monoclonais/isolamento & purificação , Camundongos , Camundongos Endogâmicos BALB C , Contagem de Ovos de Parasitas , Veia Porta/parasitologia , Proteínas Recombinantes/administração & dosagem , Proteínas Recombinantes/genética , Proteínas Recombinantes/isolamento & purificação , Esquistossomose Japônica/parasitologiaRESUMO
We have developed a novel egg yolk antibody (IgY)-coated magnetic beads antigen-capture immunoassay for detection of a circulating antigen of Schistosoma japonicum in serum samples of patients in schistosomiasis-endemic areas of China. This IgY-based immunomagnetic bead enzyme-linked immunosorbent assay (IgY-IMB-ELISA) uses polyclonal IgY-coated magnetic beads as a capture antibody, and a monoclonal IgG as a detection antibody. The sensitivity of the magnetic immunoassay was 100% (40 of 40) in cases of acute infection and 91.5% (107 of 117) in chronic cases of schistosomiasis, and no positive reaction was found in 0 of 49 healthy persons. Cross-reactivity was 3.3% (1 of 33) with clonorchiasis and 0% (0 of 20) with paragonimiasis. There was a significant correlation between ELISA absorbance value and egg count (eggs per gram feces) and a correlation coefficient of 0.88 in a small sample of 14 patients. The results demonstrated that the IgY-IMB-ELISA is a sensitive and specific assay for detection of human schistosomiasis japonica.