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Pulmonary fibrosis (PF) is a major public health issue with limited treatment options. As the active ingredient of the n-butanol extract of Amygdalus mongolica (BUT), amygdalin inhibits PF. However, its mechanisms of action are unclear and need further verification. Therefore, the purpose of the present studies was to investigate the anti-fibrotic effects of BUT on PF by serum metabolomics and the transforming growth factor β (TGF-β) pathway. Sixty male Sprague-Dawley rats were randomly divided into control, untreated PF, prednisone-treated (5 mg/kg), and BUT-treated (1.75, 1.25, 0.75 g/kg) groups, and the respective drugs were administered intragastrically for 21 days. The serum metabolomics profiles were determined by ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-QTOF/MS) and metabolism network analysis. The expression of TGF-β1, Smad-3, Smad-7, and α-smooth muscle actin (α-SMA) was measured using a real-time polymerase chain reaction in the lung tissue. BUT significantly alleviated fibrosis by reducing the mRNA expressions of TGF-β1 (from 1.73 to 1.13), Smad-3 (from 2.01 to 1.19), and α-SMA (from 2.14 to 1.19) and increasing that of Smad7 (from 0.17 to 0.62). Twenty-eight potential biomarkers associated with PF were identified. In addition, four key biomarkers were restored to baseline levels following BUT treatment, with the lowest dose showing optimal effect. Furthermore, A. mongolica BUT was found to improve PF by the pentose phosphate pathway and by taurine, hypotaurine, and arachidonic acid metabolism. These findings revealed the mechanism of A. mongolica BUT antifibrotic effects and metabolic activity in PF rats and provided the experimental basis for its clinical application.
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Objective@#To investigate the prevalence of flat feet and associated factors in school aged children in Kunming City, to provide evidence supporting the prevention of flat feet.@*Methods@#From December 2021 to February 2022, 4 444 children aged 7-13 in five primary schools in Kunming were screened for flat feet with the optical foot assessment and recording device. The incidence of flatfoot was counted, and Logistic regression was used to analyze the influencing factors of flatoccurrence.@*Results@#The overall prevalence rate was 29.10%, of which 21.79% were mild, 52.43% were moderate, 25.78% were severe, 89.10 % were bipedal, and 10.90% were monopedal. The prevalence rates in the 7-year old and 13-year old groups were 36.91% and 10.43%, respectively, and the risk in the former was 5.00 times that in the latter( OR=5.00, 95%CI =3.22-7.52). The prevalence rates in rural and urban students were 38.53%, 22.46%, respectively, and the risk in the former was 2.17 times that in the latter( OR=2.17, 95%CI =1.90-2.47). The prevalence of flat feet in male and female students were 34.21%, 23.29%, respectively, and the risk in male students was 1.71 times higher than that in female students( OR=1.71, 95%CI =1.50-1.95). The incidence of flat feet correlated with BMI, and the risk of flat feet was higher in the group with overweight and obese groups than normal( OR=1.31, 1.10, P < 0.01). @*Conclusion@#The prevalence of flat feet in school age children aged 7-13 years decreased with age. The prevalence and risk of flat feet is lower in girls than boys, and the incidence and risk of flat feet are lower in urban than rural children. The incidence of flat feet in most children is moderate, and the risk increased with increasing BMI. For school aged children with flat feet, early prevention, detection and treatment are needed.
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Objective@#To compare the efficiency and biocompatibility of four different silanes on immobilizing c(RGDfK) peptide on titanium surface.@*Methods @# After alkali-heat treatment (group OH), the titanium surface was treated with 3-aminopropyltriethoxysilane (APTES) (group OHAP), 3-chloropropyltriethoxysilane (CPTES) (group OHCP) (3-mercaptopropyltrimethoxysilane (MPTS) (group OHMPT) and 3-isobutyryloxy propyltrimethoxysilane(γ- MPS) (group OHMPS) to immobilize the c(RGDfK) cyclic peptide and constructa titanium-silane-c(RGDfK) coating. The NT group was the blank control group. The surface morphology and wettability of the coatings were detected using scanning electron microscopy and contact angle measurement. The elemental composition of the titanium surface was analyzed using X-ray photoelectron spectroscopy. After fluorescent staining with 4’,6-diamino-2-phenylindole (DAPI) and phalloidin, the adhesion of mouse preosteoblast MC3T3-E1 cells on the surface of the materials was observed using laser confocal microscopy. Cell counting kit-8 (CCK-8) and alkaline phosphatase (ALP) activity assays were used to evaluate the proliferation and osteogenic differentiation of MC3T3-E1 cells on the surface of the materials, respectively. @*Results @#Scanning electron microscope observation showed a spongy-like 3-dimensional network formed on the titanium surface after alkali-heat treatment with silane-c(RGDfK) coating adhesion. The wettability of each group was greatly improved compared to the untreated titanium surface. The element ratios of Si/Ti and amide-N/Ti in the OHMPS group were the highest. The OHAP group exhibited the best cell adhesion effect. The cell proliferation and ALP activity of the OHAP, OHMPT, and OHMPS groups were significantly higher than the control group (P <0.05); there was no statistical difference between the OHCP group and the control group.@*Conclusion @#MPTS, CPTES and γ-MPS covalently immobilized cyclic peptide c(RGDfK) on the titanium surface, which promoted adhesion, proliferation and osteogenic differentiation of MC3T3-E1 cells. Theγ-MPS conjugated C (RGDfK)cyclic peptide exhibited the best effect. MPTS, CPTES and γ-MPS coupled with c(RGDfK) cyclic peptides had similar biological properties.
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In order to study the biological function of pig BST-2 gene,the BST-2 gene was amplified with specific primers from porcine kidney tissue,and molecular characterization of BST-2 nuclectide and amino acid sequence were analyzed with bioinformatics tools and online server.Then the prokaryotic expression and tissue expression profile analysis was carried out.The results showed that the full length of pig BST-2 gene was 851 bp and contained 23 bp of 5'-UTR,294 bp of 3'-UTR and 534 bp of CDS and the gene encoded 177 aa.Amino acid sequence analysis of pig BST-2 protein showed 46.1％ identity with gorilla gorilla,41.7％ with cricetulus griseus,39.5％ with mus musculus,35.4％ with equus asinus,42.0％ with felis catus,40.5％ with bos mutus,44.4％ with macaca mulatta,38.7％ with ovis aries and 46.8％ with homo sapiens.BST-2 protein contained 2 transmembrane structure (27-49 aa and 154-176 aa),2 glycosylation sites and 14 potential phosphorylation sites including ATM,CK Ⅱ,PKA,PKC binding sites.The pig BST-2 protein was expressed in Vero cells after translated the recombinant plasmid FLAG-BST-2.Semiquantitative PCR results showed that BST-2 gene was expressed in all the tissues,especially in lymph nodes,thymus,tonsils,spleen,large intestine and small intestine.This study provide a foundation for further understanding the antiviral mechanism of pig BST-2 protein.
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Objective To analyze the correlation between the TCM diagnosis and treatment data to provide support for scientific research and clinical treatment.Methods The characteristics of TCM diagnosis and treatment data were analyzed.An improved Apriori algorithm with grouping association was put forward,and association analysis on the data from the encephalopathy database of some hospital was carried out to verify the feasibility of the algorithm.Results Grouped operation was executed for association properties to reduce the association between noncorrelated data,so that the efficiency of the algorithm was enhanced greatly.Conclusion Improved Apriori algorithm with grouping association consumes shorter time than the classical one,and thus is worthy promoting in TCM diagnosis and treatment data application.
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Objective To analyze the correlation between the TCM diagnosis and treatment data to provide support for scientific research and clinical treatment.Methods The characteristics of TCM diagnosis and treatment data were analyzed.An improved Apriori algorithm with grouping association was put forward,and association analysis on the data from the encephalopathy database of some hospital was carried out to verify the feasibility of the algorithm.Results Grouped operation was executed for association properties to reduce the association between noncorrelated data,so that the efficiency of the algorithm was enhanced greatly.Conclusion Improved Apriori algorithm with grouping association consumes shorter time than the classical one,and thus is worthy promoting in TCM diagnosis and treatment data application.
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Objective To investigate the status of syphilis and HCV infection and the influencing factors among community drug addicts in Jiaxing City.Methods HCV and syphilis were detected among community drug addicts in Jiaxing City from 2014-2015 and a questionnaire survey were conducted.Influencing factors of syphilis and HCV was analyzed in this study.Results A total of 449 drug users including 356 males (79.29％) and 93 females (20.71％) were investigated,with the age ranged from 16 to 47 and averaged 27.50 ± 12.28.A total of 370 of them take addictive drugs (82.41％),including 42 of them take more than 2 kinds of drugs.15 cases of Syphihs (double positive) were found,and the positive rate was 3.34％.There were significant difference among community drug addicts with different education level (P ＜0.05).The positive rate of HCV among males were lower than that of females (P ＜ 0.05),and the positive rate of HCV among heroin addicts were higher than other drug addicts (P ＜ 0.05).The positive rate of HCV of injection drug users were higher than oral drug addicts(OR =17.341,95％ CI:8.387-35.857,P ＜ 0.01).There was associations among condom use with double positive syphilis and HCV (OR =0.210,95％ CI:0.064-0.689,P ＜0.01;OR =0.131,95％ CI:0.063 -0.273,P ＜ 0.01),respectively.Conclusion Community drug addics are mainly young adults,drug injection and not using condom may increase the risks of TP and HCV infection.
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Swine influenza viruses (SwIVs) cause considerable morbidity and mortality in domestic pigs, resulting in a significant economic burden. Moreover, pigs have been considered to be a possible mixing vessel in which novel strains loom. Here, we developed and evaluated a novel M2e-multiple antigenic peptide (M2e-MAP) as a supplemental antigen for inactivated H3N2 vaccine to provide cross-protection against two main subtypes of SwIVs, H1N1 and H3N2. The novel tetra-branched MAP was constructed by fusing four copies of M2e to one copy of foreign T helper cell epitopes. A high-yield reassortant H3N2 virus was generated by plasmid based reverse genetics. The efficacy of the novel H3N2 inactivated vaccines with or without M2e-MAP supplementation was evaluated in a mouse model. M2e-MAP conjugated vaccine induced strong antibody responses in mice. Complete protection against the heterologous swine H1N1 virus was observed in mice vaccinated with M2e-MAP combined vaccine. Moreover, this novel peptide confers protection against lethal challenge of A/Puerto Rico/8/34 (H1N1). Taken together, our results suggest the combined immunization of reassortant inactivated H3N2 vaccine and the novel M2e-MAP provided cross-protection against swine and human viruses and may serve as a promising approach for influenza vaccine development.
Subject(s)
Animals , Female , Mice , Antibodies, Viral/blood , Antigens, Viral/genetics , Body Weight , Cross Protection/immunology , Disease Models, Animal , Epitopes, T-Lymphocyte/genetics , Influenza A Virus, H3N2 Subtype/genetics , Influenza Vaccines/immunology , Mice, Inbred BALB C , Orthomyxoviridae Infections/immunology , Peptides/genetics , Random Allocation , Survival Analysis , Vaccines, Synthetic/immunology , Virus ReplicationABSTRACT
Inhibitors of kinesin spindle protein (KSP) are a promising class of anticancer agents that cause mitotic arrest and induce apoptosis of tumor cells. A series of novel tetrahydro-beta-carboline derivatives were synthesized as kinesin spindle protein inhibitor and evaluated as potential antitumor agents. All compounds showed promising KSP inhibitiory activity. Compounds 8 and 9 exhibited better antitumor activity (Lung/A549, Stomach/AGS) than CK0106023 with GI50/IC50 values (1.07/1.62 and 1.46/3.27 micromol x L(-1), 1.09/>10 and 1.22/6.33 micromol x L(-1), respectively).
Subject(s)
Humans , Antineoplastic Agents , Chemistry , Pharmacology , Carbolines , Chemistry , Pharmacology , Cell Line, Tumor , Cell Proliferation , Inhibitory Concentration 50 , Kinesins , Pharmacology , Molecular StructureABSTRACT
Establishment of recombinant porcine reproductive and respiratory syndrome virus (PRRSV) with co-expression E2 Epitope of Classical Swine Fever virus (CSFV) is a crucial step to develop a genetic engineered vaccine against PRRSV and CSFV. Reverse genetic manipulation could be adopted as a com monly used technique. In this study, we focus on using nonessential regions of NSP2 (aa480-532 and aa508-532) as viral vector to express E2 Epitope of CSFV. A neutralizing epitope of classical swine fever virus (CSFV) E2 protein was inserted into the two nonessential region of nsp2 by the method of mutant PCR, basing on the infectious clone of HuN4-F112 vaccine strain. The co-expressed full-length cDNA clones (psk-HuN4-F112-delta508-532 + E2 and psk-HuN4-F112-delta480-532 + E2) were assembled by cloning and splice of the gene fragments. The completely assembled full-length cDNA clones were confirmed by sequence and Swa I enzyme digestion. Capped RNAs were transcribed in vitro from a full-length cDNA clone of the viral genome and transfected into BHK-21 cells by liposome to acquire the rescued virus. The rescued recombinant viruses were passaged on MARC-145 cells. The successfully rescued viruses were tested by RT-PCR, digestion, and genome sequence. The results showed that these rescued viruses could be distinguished from the parental virus (HuN4-F112) with the mutant genetic marker (Mlu I enzyme site of virual genome at 14 667nt was created by synonymous mutation) and the inserted nsp2 gene region. The results of IFA showed that the inserted E2 epitope could be expressed by the recombinant viruses and the E2 epitope gene was stable during the viral serial passage. The results of plaque assay and viral growth curve showed that the recovery viruses possessed similar characterses of viral growth to those of the parental virus. In summary, the full-length infectious cDNA clones containing the marker gene were constructed and the marker recombinant viruses were rescued. The results suggested that these stable infectious clones could be used as an important tool for development of novel vaccine against PRRSV.
Subject(s)
Base Sequence , Cysteine Endopeptidases , Genetics , Epitopes , Genetics , Molecular Sequence Data , Porcine respiratory and reproductive syndrome virus , Genetics , Viral Envelope Proteins , Genetics , Allergy and Immunology , Viral Vaccines , Allergy and ImmunologyABSTRACT
Using mutation PCR, we cloned the target gene containing 421-480nt (141-160aa) and 598-639nt (200-213aa) of VP1 gene of foot and mouth disease virus (FMDV) into the deleted region (508-532aa) of Nsp2 gene of a highly pathogenic porcine reproductive and respiratory syndrome virus derived vaccine strain (HuN4-F112) that was used as vector. The recombinant cDNA was in vitro transcribed followed by transfection of BHK-21 cells for 36 h. Then, the supernatant of the cell culture was continuously seeded to monolayer of MARC-145 cells for recovery of the recombinant virus. CPE was obviously visible after a couple of passages in the seeded MARC-145, and the rescued virus (designated as rPRRSV-F112-O/VP1ep) was identified by Mlu I digestion, sequencing and immunofluorescence assay. Meanwhile, expression of inserted FMDV epitopes was also detected by indirect immunofluorescence assay with polyclonal antibodies against VP1 protein of FMDV. The analysis of biological characteristics shows that the titer of the rescued recombinant PRRSV (TCID50 = -log10(-6.75)/0.1 mL) was similar to its direct parental virus rHuN4-F112-delta508-532, but higher than rHuN4-F112.
Subject(s)
Animals , Antigens, Viral , Allergy and Immunology , Base Sequence , Capsid Proteins , Allergy and Immunology , Cell Line , Cysteine Endopeptidases , Genetics , Epitopes , Genetics , Foot-and-Mouth Disease , Allergy and Immunology , Foot-and-Mouth Disease Virus , Genetics , Allergy and Immunology , Molecular Sequence Data , Mutation , Porcine respiratory and reproductive syndrome virus , Genetics , Allergy and Immunology , Recombination, Genetic , Swine , Transfection , Vaccines, Attenuated , Genetics , Allergy and Immunology , Viral Envelope Proteins , Genetics , Allergy and Immunology , Viral Vaccines , Genetics , Allergy and ImmunologyABSTRACT
Background Recombinant hirudin variant Ⅲ(rHV3) can effectively prevent galactose-induced human lens epithelial cells LECs injury,but little is known about the molecular mechanism of its action.Objective The present study was to investigate the effects of rHV3 on the expression of apoptosis-related genes in damaged LECs induced by galactose.Methods The rHV3 was extracted by our research group,and the biological activity of rHV3 was identified by titration of thrombase according to Markwardt's method.Human LECs (SRA01/04) were cultured using 125×10-3 mol/L D-galactose+10% FBS+D/F12 medium to establish the damaged human LECs model.rHV3 was added into the medium of the damaged human LECs model.Human LECs were cultured in D/F12 medium containing 10% FBS as normal control.The expression of apoptosis-related genes,such as aldose reductase (AR),bax,bcl2 and p53,in LECs at the mRNA level was detected using RT-PCR.The abundance ratio of target genes was presented with the absorbance (A) of gene mRNA/GAPDH mRNA.Results Compared to the normal control group,the A values of AR mRNA/GAPDH mRNA,bax mRNA/GAPDH mRNA and p53 mRNA/GAPDH mRNA were significantly elevated in model group (t=3.90E-06,t=8.44E-04,t=5.15E-08,P＜0.01).However,in the rHV3-treated group,the A values of AR mRNA/GAPDH mRNA,bax mRNA/GAPDH mRNA and p53 mRNA/GAPDH mRNA were lower than those of model group (t=5.90E-06,t=1.51E-04,t=3.42E-06,P＜0.01).The bcl2 mRNA/GAPDH mRNA was markedly downregulated in the model group when compared with the normal control group (t=1.86E-05,P＜0.01);while after rHV3 addition,bcl2 mRNA/GAPDH mRNA increased in comparison with the model group (t=8.56E-05,P＜0.01).Conclusion 125×10-3mol/L D-galactose induces the damage and apoptosis of human LECs.rHV3 likely plays a protective function on D-galactose-induced damage of human LECs by inhibiting the polyol pathway and mitochondria-mediated pathway.
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The aim is to determine the primary structure of a new hirudin and reteplase fusion protein (HV12p-rPA) by LC-ESI-MS/MS spectrometry. The molecular weight of the hirudin and reteplase fusion protein (HV12p-rPA) was measured by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). The HV12p-rPA was digested with trypsin and chymotrypsin separately and the peptides in the digest mixtures were identified by LC-ESI-MS/MS. The molecular weight of HV12p-rPA was 41,472 Da, which was in accordance with the theoretical value. The peptide fragments of HV12p-rPA digested with trypsin were identified by LC-ESI-MS/MS spectrometry and the results indicated that the fusion protein contained r-PA. Then, the peptides of HV12p-rPA digested with chymotrypsin were identified by the same method. The results indicated that the fusion protein contained HV12p and the linker-containing peptide, DEGGGSY. MASDF and LDWIRDNMRP were identified as the N-terminal and C-terminal containing peptides in the chymotryptic digest mixture of the fusion protein. All of the Xcorr values exceeded 1.5, some of which were above 3.0, showing that the results were correct and credible and a sequence coverage of 85% was achieved. HPLC/MS analysis coupled with uncompleted digestion indicated that all these peptides were arranged with the correct order as expected. Thus, sequence of the fusion protein was confirmed and it was consistent with our design in upstream construction.
Subject(s)
Amino Acid Sequence , Chromatography, Liquid , Methods , Chymotrypsin , Chemistry , Fibrinolytic Agents , Chemistry , Hirudins , Chemistry , Molecular Sequence Data , Molecular Weight , Peptide Fragments , Recombinant Fusion Proteins , Chemistry , Recombinant Proteins , Chemistry , Spectrometry, Mass, Electrospray Ionization , Methods , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization , Methods , Tandem Mass Spectrometry , Methods , Tissue Plasminogen Activator , Chemistry , Trypsin , ChemistryABSTRACT
Kinesin spindle protein (KSP/Eg5) is essential for the formation and maintenance of bipolar spindles during mitosis. Inhibition of this protein leads to cell cycle arrest and apoptosis without interfering other microtubule-dependent processes. Therefore, it is a potential target in cancer therapy. Here, a series of tetrahydro-beta-carboline derivatives 5a - k were synthesized as kinesin spindle protein inhibitor. Their structures were confirmed with 1H NMR, ESI-MS and elemental analysis. The synthesized compounds were evaluated for their inhibition of KSP.
Subject(s)
Antineoplastic Agents , Chemistry , Pharmacology , Carbolines , Chemistry , Pharmacology , Kinesins , Metabolism , Molecular StructureABSTRACT
Application of HPLC-ESI-ITMS in the quality control of carboxyterminal sequence confirmation for insulin and insulin chain B was studied. The solution of intact insulin or insulin chain B was added to the solution of carboxypeptidase P (CPP) and carboxypeptidase Y (CPY). Fractions of appropriate volume were removed at some appointed time points, acidified with the same amount of 1% formic acid to stop the digestion, and then briefly vortexed for HPLC-ESI-ITMS analysis. Mobile phase A consisted of 0.02% TFA in 98% ultra-pure water and 2% acetonitrile. Mobile phase B consisted of 0.02% TFA in 98% acetonitrile and 2% ultra-pure water. The solution used for post-column fix consisted of propionic acid and isopropyl alcohol (20 : 80, v/v). Chromatographic separation was carried out on a reversed-phase column (Zorbax Prosphere C18, 300A, 5 microm, 2.1 mm ID x 150 mm length). The molecular weights of the multiply charged ions representing consecutive truncated losses of carboxyterminal amino acids were determined by the use of HPLC-ESI-ITMS. The differences between the consecutive truncated peptides are the experimental weights of the carboxyterminal amino acid residues. The carboxyterminal amino acid residue Ala, which released from chain B of intact insulin, was confirmed in the nanomolar concentration range by analyzing the molecular weight of the truncated peptides. Another one carboxyterminal amino acid Ala was confirmed in the nanomolar concentration range of insulin chain B. In the quality control for recombinant DNA product or natural protein, the confirmation of 1 - 3 carboxyterminal amino acid residues is regarded to be up to standard. One amino acid residue of insulin or insulin chain B could be confirmed accurately in the nanomolar concentration range. The results showed that intact insulin could be directly sequenced in the quality control without separating chain B from chain A. There would be no need to separate chain A from chain B to identify carboxyterminal of intact insulin. Furthermore, the method saved us a lot of trouble from the preparation and purification of insulin chain A and chain B.
Subject(s)
Amino Acid Sequence , Carboxypeptidases , Chemistry , Cathepsin A , Chemistry , Chromatography, High Pressure Liquid , Methods , Insulin , Chemistry , Reference Standards , Molecular Sequence Data , Molecular Weight , Peptide Fragments , Chemistry , Quality Control , Spectrometry, Mass, Electrospray Ionization , MethodsABSTRACT
<p><b>AIM</b>To investigate the protective effects of shark hepatic stimulator substance (sHSS) against acute hepatic injury induced by acetaminophen (AAP) in mice.</p><p><b>METHODS</b>Acute hepatic injury model of Balb/c mice was induced by a single intraperitoneal injection of AAP (200 mg.kg-1, i.p.). Serum ALT and AST activities were analyzed. The changes of microstructure and ultrastructure of hepatocyte were observed under optical and electronic microscope. The hepatocyte apoptosis was analyzed by flow cytometer and the expression level of Fas mRNA was determined by RT-PCR.</p><p><b>RESULTS</b>The activities of serum ALT and AST were significantly decreased and both necrosis and inflammatory infiltration were improved in the mice treated with sHSS 3.0 and 1.5 mg.kg-1. sHSS (3.0 mg.kg-1) prevented the ultrastructural changes of hepatocytes caused by AAP, decreased the percentage of apoptotic cells, and downregulated the expression level of Fas mRNA.</p><p><b>CONCLUSION</b>sHSS protected hepatocytes from AAP-induced injury, which might be associated with its protection of the mitochondria and inhibition of apoptosis and expression of Fas mRNA in hepatocytes.</p>
Subject(s)
Animals , Female , Mice , Acetaminophen , Apoptosis , Chemical and Drug Induced Liver Injury , Pathology , Growth Substances , Pharmacology , Mice, Inbred BALB C , Peptides , Pharmacology , Protective Agents , Pharmacology , RNA, Messenger , Genetics , Random Allocation , Sharks , fas Receptor , GeneticsABSTRACT
<p><b>AIM</b>To establish antibody sandwich enzyme-linked immunoadsorbent assay for determination of recombinant E. coli L-asparaginase in rat plasma and study its pharmacokinetics.</p><p><b>METHODS</b>A Japanese white rabbit was immunized with recombinant E. coli L-asparaginase. Immunoglobulin G was separated and purified by using DEAE-cellulose chromatography. Conjugation of horseradish peroxidase to immunoglobulin G was obtained using the two-step glutaraldehyde method. Recombinant E. coli L-asparaginase protein in plasma was measured by antibody sandwich enzyme-linked immunoadsorbent assay. Pharmacokinetic parameters were assessed with model-dependent method.</p><p><b>RESULTS</b>The linearities was 1-64 U.L-1. Concentration-time profile after i.v. of 1.25, 2.50, 5.00 kU.kg-1 of recombinant E. coli L-asparaginase fitted with a two-compartment model. The first and terminal elimination T1/2 were 0.50-0.57 h and 2.45-3.02 h, respectively. The AUC was linearly related to the doses.</p><p><b>CONCLUSION</b>Antibody sandwich enzyme-linked immunoadsorbent assay was constant, reliable, sensitive, and suitable for the determination of recombinant L-asparaginase. Pharmacokinetics of recombinant E. coli L-asparaginase in rats is warranted for the design of future clinical trails.</p>