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1.
Chinese Herbal Medicines ; (4): 316-325, 2020.
Article in Chinese | WPRIM | ID: wpr-842002

ABSTRACT

Objective: The research indicated that the nature of Chinese medicine is mainly related to body's substance and energy metabolism. The purpose of the study is to elucidate the substance basis for warm nature of Poria cocos (called Fuling (FL) in Chinese). Methods: In terms of the effects of its separated fractions on the substance and energy metabolism in rat models of cold-deficiency with Aconiti Lateralis Radix Praeparata (called Fuzi (FZ) in Chinese), with hot nature, as reference drug. Biochemical indexes in the material metabolism, energy metabolism, endocrine system, nervous system and nucleotide system were determined, then analyzed by additive, cluster and principal component analysis (PCA). Results: The medicinal natures of oligosaccharides and amino acids fractions were attributable to plain and crude polysaccharides, volatile oils and triterpenoids fractions were attributable to mild warm. Conclusion: The nature of FL was regarded as mild warm based on the old records of Chinese medicine and fractions of crude polysaccharides, volatile oils and triterpenoids might be the main substance basis for the warm nature of FL. It is the first time that substance basis of FL was elucidated from view point of medicinal nature.

2.
Chinese Medical Journal ; (24): 1015-1024, 2020.
Article in English | WPRIM | ID: wpr-827709

ABSTRACT

BACKGROUND@#Human infections with zoonotic coronaviruses (CoVs), including severe acute respiratory syndrome (SARS)-CoV and Middle East respiratory syndrome (MERS)-CoV, have raised great public health concern globally. Here, we report a novel bat-origin CoV causing severe and fatal pneumonia in humans.@*METHODS@#We collected clinical data and bronchoalveolar lavage (BAL) specimens from five patients with severe pneumonia from Wuhan Jinyintan Hospital, Hubei province, China. Nucleic acids of the BAL were extracted and subjected to next-generation sequencing. Virus isolation was carried out, and maximum-likelihood phylogenetic trees were constructed.@*RESULTS@#Five patients hospitalized from December 18 to December 29, 2019 presented with fever, cough, and dyspnea accompanied by complications of acute respiratory distress syndrome. Chest radiography revealed diffuse opacities and consolidation. One of these patients died. Sequence results revealed the presence of a previously unknown β-CoV strain in all five patients, with 99.8% to 99.9% nucleotide identities among the isolates. These isolates showed 79.0% nucleotide identity with the sequence of SARS-CoV (GenBank NC_004718) and 51.8% identity with the sequence of MERS-CoV (GenBank NC_019843). The virus is phylogenetically closest to a bat SARS-like CoV (SL-ZC45, GenBank MG772933) with 87.6% to 87.7% nucleotide identity, but is in a separate clade. Moreover, these viruses have a single intact open reading frame gene 8, as a further indicator of bat-origin CoVs. However, the amino acid sequence of the tentative receptor-binding domain resembles that of SARS-CoV, indicating that these viruses might use the same receptor.@*CONCLUSION@#A novel bat-borne CoV was identified that is associated with severe and fatal respiratory disease in humans.


Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Betacoronavirus , Genetics , Coronavirus Infections , Diagnostic Imaging , Therapeutics , Virology , Pandemics , Pneumonia, Viral , Diagnostic Imaging , Therapeutics , Virology , Tomography, X-Ray , Treatment Outcome
3.
Journal of Acupuncture and Tuina Science ; (6): 53-58, 2020.
Article in Chinese | WPRIM | ID: wpr-824951

ABSTRACT

Objective: To observe the therapeutic efficacy of tuina plus cupping on chronic fatigue syndrome (CFS). Methods: A total of 100 CFS patients meeting the inclusion criteria were randomly divided into a treatment group and a control group according to the random number table in the sequence of visit, with 50 patients in each group. The treatment group was treated with tuina plus cupping therapy, 3 times a week. The control group was given oral tablets of wild American ginseng lozenges. After 3 months of treatment, the score of fatigue questionnaire scale (FS-14) was observed and the efficacy was evaluated. Results: The total effective rate was 92.0% in the treatment group, versus 76.0% in the control group and the difference between the two groups was statistically significant (P<0.05). After treatment, the FS-14 scores of the two groups decreased, and the intra-group differences were statistically significant (both P<0.05). The change of FS-14 score after treatment in the treatment group was more significant than that in the control group (P<0.05). Conclusion: Tuina plus cupping is effective for CFS, and can improve fatigue symptoms significantly.

4.
Chinese Medical Journal ; (24): E001-E001, 2020.
Article in English | WPRIM | ID: wpr-817253

ABSTRACT

Background: Human infections with zoonotic coronaviruses (CoVs), including severe acute respiratory syndrome (SARS)-CoV and Middle East respiratory syndrome (MERS)-CoV, have raised great public health concern globally. Here, we report a novel bat-origin CoV causing severe and fatal pneumonia in humans. Methods: We collected clinical data and bronchoalveolar lavage (BAL) specimens from five patients with severe pneumonia from Jin Yin-tan Hospital, Wuhan, Hubei province, China. Nucleic acids of the BAL were extracted and subjected to next-generation sequencing. Virus isolation was carried out, and maximum-likelihood phylogenetic trees were constructed. Results: Five patients hospitalized from December 18 to December 29, 2019 presented with fever, cough, and dyspnea accompanied by complications of acute respiratory distress syndrome. Chest radiography revealed diffuse opacities and consolidation. One of these patients died. Sequence results revealed the presence of a previously unknown β-CoV strain in all five patients, with 99.8–99.9% nucleotide identities among the isolates. These isolates showed 79.0% nucleotide identity with the sequence of SARS-CoV (GenBank NC_004718) and 51.8% identity with the sequence of MERS-CoV (GenBank NC_019843). The virus is phylogenetically closest to a bat SARS-like CoV (SL-ZC45, GenBank MG772933) with 87.6–87.7% nucleotide identity, but is in a separate clade. Moreover, these viruses have a single intact open reading frame gene 8, as a further indicator of bat-origin CoVs. However, the amino acid sequence of the tentative receptor-binding domain resembles that of SARS-CoV, indicating that these viruses might use the same receptor. Conclusion: A novel bat-borne CoV was identified that is associated with severe and fatal respiratory disease in humans.

5.
Journal of Forensic Medicine ; (6): 672-676, 2020.
Article in Chinese | WPRIM | ID: wpr-985164

ABSTRACT

Objective To explore the correlation between 4 tag single nucleotide polymorphisms (Tag SNP) sites (rs7721799, rs32897, rs7718461, rs10062367) of corticotropin releasing hormone binding protein (CRHBP) and schizophrenia and aggressive behavior in the Yunnan Han population. Methods Case-control correlation analysis was used to establish a complex amplification system. Improved multiplex ligase detection reaction (iMLDR) technology was used to detect the genotypes of 4 SNP sites of CRHBP gene of 163 Han schizophrenic patients (including 81 patients with aggressive behavior, 82 patients without aggressive behavior) and 345 healthy Han individuals, which were analyzed statistically by SPSS 19.0, Haploview 4.2 and PHASE 2.1 software. Results There was no correlation between the 3 SNP sites of CRHBP gene and the onset of schizophrenia except for the rs7718461 site (P>0.05). The relative risk of aggressive behavior of patients carrying GG or GA genotype at rs7718461 site were 4.903 times higher than those carrying AA genotype (P<0.05). Conclusion The CRHBP gene may not be associated with the occurrence of schizophrenia in Yunnan Han population, but AA genotype of rs7718461 may reduce the risk of aggressive behavior in schizophrenia patients.


Subject(s)
Humans , Asian People/genetics , Carrier Proteins/genetics , China , Gene Frequency , Genetic Predisposition to Disease , Genotype , Polymorphism, Single Nucleotide , Schizophrenia/genetics
6.
International Eye Science ; (12): 2022-2025, 2019.
Article in Chinese | WPRIM | ID: wpr-756825

ABSTRACT

@#AIM: To evaluate hesperidin's inhibitory effect on the proliferation of human pterygium fibroblasts(HPF)cultured <i>in vitro</i> and its influence on the expression of cyclin D.<p>METHODS: The fresh tissue of human pterygium was cultivated by adherent cell culture <i>in vitro </i>and adherent cells were appraised by immune fluorescence staining. HPF cells were treated with hesperidin(24μmol/L, 48μmol/L, 64μmol/L, 72μmol/L, 96μmol/L, 120μmol/L)and MMC(1.5μmol/L, 7.5μmol/L and 30.0μmol/L). The inhibition rate of cell proliferation was detected by MTT assay 24h, 48h and 72h after treatment, and appropriate concentration and time were selected. The relative expression of cyclin D in HPF was detected by Western blot.<p>RESULTS: When HPF were treated respectively with hesperidin(48μmol/L, 72μmol/L)and MMC(7.5μmol/L)for 48h, Western blot results showed the relative expressions of cyclin D in blank control group(normal culture), MMC group, hesperidin(48μmol/L)group and hesperidin(72μmol/L)group to be 1.20±0.02, 0.60±0.03, 0.54±0.02, 0.45±0.07(<i>F</i>=73.025, <i>P</i>=0.001)respectively. The relative expressions of cyclin D in MMC group and hesperidin group were lower than that of blank control group(<i>P</i><0.05); while the relative expressions of cyclin D in MMC group and hesperidin(48μmol/L, 72μmol/L)group showed no significant difference(<i>P</i>>0.05).<p>CONCLUSION:Hesperidin can inhibit the proliferation of HPF by reducing the relative expression of cyclin D.

7.
China Journal of Chinese Materia Medica ; (24): 4519-4527, 2018.
Article in Chinese | WPRIM | ID: wpr-771584

ABSTRACT

This present study was to investigate the metabolism and excretion of characteristic polyphenols such as flavonoids and coumarins in urine and feces of rats after intragastric administration of ethanol extracts of Citri Reticulatae Pericarpium Viride and Citri Reticulatae Pericarpium. The urine and feces of rats were collected after intragastric administration of 70% ethanol extracts of Citri Reticulatae Pericarpium Viride and Citri Reticulatae Pericarpium. Rapid resolution liquid chromatography coupled with quadrupole tandem mass spectrometry (RRLC-QqQ-MSn) was applied to compare the contents of polyphenols in ethanol extract, urine and feces. By comparing with reference substance, 30 polyphenols were identified from the ethanol extracts of Citri Reticulatae Pericarpium Viride and Citri Reticulatae Pericarpium, including flavone glycosides, flavones, flavonone glycosides, flavonones, flavonol glycosides, polymethoxyflavones, coumarins, and limonoids and so on. The detection of various types of compounds showed differences in contents between the intestinal metabolism and excretion in the feces after systemic circulatory metabolism and renal excretion. The results showed that the polymethoxyflavones and flavonones were primarily excreted through urine, and the flavonone glycosides and limonoids were primarily excreted through feces. However, coumarins were hardly detected in feces and urine, indicating that coumarins may be metabolized in the body.


Subject(s)
Animals , Rats , Citrus , Drugs, Chinese Herbal , Feces , Flavonoids , Tandem Mass Spectrometry
8.
Journal of Experimental Hematology ; (6): 1116-1121, 2018.
Article in Chinese | WPRIM | ID: wpr-689519

ABSTRACT

<p><b>OBJECTIVE</b>To study the effect of HDAC inhibitor Scriptaid on multiple myeloma IM9 cells and preliminarily clarify the mechanism of Scriptaid-induced cell apoptosis.</p><p><b>METHODS</b>The cell viability, cell cycle and cell apoptosis were measured by CCK8 assay and flow cytometry respectively, the relative target gene expression levels were detected by RT-PCR, the effect of Scriptaid on p21 promoter activity was detected by using luciferase reporter assay.</p><p><b>RESULTS</b>Scriptaid inhibited IM9 cell viability in a dose-dependent manner. Scriptaid induced IM9 cell cycle arrest at G/M phase in a dose-dependent manner. Scriptaid triggered IM9 cell apoptosis was obviously, the mRNA levels of apoptosis-related proteins Caspase 9, Caspase 3 and PARP1 were also activated. The apoptosis-associated factors BAD, PTEN and p21 increased following treatment with different dose of Scriptaid, meanwhile, p21 promoter activity was also activated significantly.</p><p><b>CONCLUSION</b>HDAC inhibitor Scriptaid can promote IM9 cell apoptosis by transcriptional activation of p21 promoter in concentration-dependent manner.</p>


Subject(s)
Humans , Apoptosis , Cell Cycle , Cell Line, Tumor , Cell Proliferation , Cyclin-Dependent Kinase Inhibitor p21 , Histone Deacetylase Inhibitors , Pharmacology , Hydroxylamines , Pharmacology , Quinolines , Pharmacology
9.
Chinese Journal of Analytical Chemistry ; (12): 1448-1454, 2017.
Article in Chinese | WPRIM | ID: wpr-662280

ABSTRACT

By detecting SRY gene of cell-free fetal DNA ( cffDNA) in maternal peripheral blood, the sex of fetuses was determined, the risks of sex-linked genetic disorders were assessed and the birth rate of sick fetuses was decreased. A method of real-time polymerase chain reaction ( PCR) coupled with invader assay was established to detect SRY gene. This method possessed the advantages such as high sensitivity, high specificity and non-contaminated with closed tube detection. Under the optimized reaction conditions such as 250 nmol/L detection probes, 7. 5 U FEN1 enzyme, 0. 5 U Taq polymerase and 67℃ of annealing temperature in pre-amplification, the simulated samples as low as 4% ( 4 copies/μL ) were detected and two clinical samples with the gestation age of 9 weeks and 10 weeks were successfully detected. The detection results showed that this method could be used to detect SRY gene of cffDNA in maternal peripheral blood, providing an effective technique for clinical non-invasive prenatal diagnosis based on SRY gene.

10.
Chinese Journal of Analytical Chemistry ; (12): 1448-1454, 2017.
Article in Chinese | WPRIM | ID: wpr-659720

ABSTRACT

By detecting SRY gene of cell-free fetal DNA ( cffDNA) in maternal peripheral blood, the sex of fetuses was determined, the risks of sex-linked genetic disorders were assessed and the birth rate of sick fetuses was decreased. A method of real-time polymerase chain reaction ( PCR) coupled with invader assay was established to detect SRY gene. This method possessed the advantages such as high sensitivity, high specificity and non-contaminated with closed tube detection. Under the optimized reaction conditions such as 250 nmol/L detection probes, 7. 5 U FEN1 enzyme, 0. 5 U Taq polymerase and 67℃ of annealing temperature in pre-amplification, the simulated samples as low as 4% ( 4 copies/μL ) were detected and two clinical samples with the gestation age of 9 weeks and 10 weeks were successfully detected. The detection results showed that this method could be used to detect SRY gene of cffDNA in maternal peripheral blood, providing an effective technique for clinical non-invasive prenatal diagnosis based on SRY gene.

11.
Biomedical and Environmental Sciences ; (12): 68-74, 2017.
Article in English | WPRIM | ID: wpr-296514

ABSTRACT

The highly pathogenic avian influenza (HPAI) H5N1 virus has caused several outbreaks in domestic poultry. Despite great efforts to control the spread of this virus, it continues to evolve and poses a substantial threat to public health because of a high mortality rate. In this study, we sequenced whole genomes of eight H5N1 avian influenza viruses isolated from domestic poultry in eastern China and compared them with those of typical influenza virus strains. Phylogenetic analyses showed that all eight genomes belonged to clade 2.3.2.1 and clade 7.2, the two main circulating clades in China. Viruses that clustered in clade 2.3.2.1 shared a high degree of homology with H5N1 isolates located in eastern Asian. Isolates that clustered in clade 7.2 were found to circulate throughout China, with an east-to-west density gradient. Pathogenicity studies in mice showed that these isolates replicate in the lungs, and clade 2.3.2.1 viruses exhibit a notably higher degree of virulence compared to clade 7.2 viruses. Our results contribute to the elucidation of the biological characterization and pathogenicity of HPAI H5N1 viruses.


Subject(s)
Animals , China , Influenza A Virus, H5N1 Subtype , Genetics , Virulence , Influenza in Birds , Virology , Mice, Inbred BALB C , Phylogeny , Poultry
12.
Chinese Journal of Oncology ; (12): 790-792, 2008.
Article in Chinese | WPRIM | ID: wpr-357336

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the cause and treatment as well as prevention measures of rarely occurring severe complications after transcatheter arterial chemoembolization (TACE) for primary hepatic carcinoma.</p><p><b>METHODS</b>573 consecutive patients with primary hepatic carcinoma underwent a total of 1252 TACE procedures from January 2005 to July 2007. All the patients who developed complications after TACE received imaging and biochemical examinations. The cause, treatment and preventive measures of the complications in the 573 cases were analyzed.</p><p><b>RESULTS</b>There were upper gastrointestinal hemorrhage in 3 cases, hepatic failure in 4, pulmonary embolism in 1, cholecystitis in 4, hepatic encephalopathy in 2, gastric perforation in 1, and intrahepatic biloma in 2 cases. Two patients died of the complications: 1 of hepatic failure and 1 of gastric perforation.</p><p><b>CONCLUSION</b>The rarely occurring severe complications after transcatheter arterial chemoembolization for primary hepatic carcinoma is correlated with poor hepatic function and portal hypertension before therapy, overdose and reflux of chemotherapeutic agents or allotopic chemoembolism, etc. It can be reduced or prevented through careful selection of proper cases before the treatment, close observation, and protection of hepatic function and gastric mucosa after treatment.</p>


Subject(s)
Adolescent , Adult , Aged , Aged, 80 and over , Female , Humans , Male , Middle Aged , Young Adult , Antineoplastic Combined Chemotherapy Protocols , Therapeutic Uses , Carcinoma, Hepatocellular , Therapeutics , Chemoembolization, Therapeutic , Methods , Epirubicin , Fluorouracil , Gastrointestinal Hemorrhage , Hepatic Encephalopathy , Iodized Oil , Liver Failure , Liver Neoplasms , Therapeutics , Mitomycin , Pulmonary Embolism
13.
Chinese Journal of Industrial Hygiene and Occupational Diseases ; (12): 725-728, 2008.
Article in Chinese | WPRIM | ID: wpr-347230

ABSTRACT

<p><b>OBJECTIVE</b>To study the effects of domoic acid (DA) on membrane function of primary cultured rat glial cell.</p><p><b>METHODS</b>After the glial cells were treated with 6.4 x 10(-2), 6.4 x 10(-3) and 6.4 x 10(-4) micromol/L DA for 24 h, the activities of Na(+)-K(+)-ATPase and Ca(2+)-Mg(2+)-ATPase, the membrane fluidity and the permeability were measured to reflect the membrane function.</p><p><b>RESULTS</b>After treatment of DA for 24 h, the activities of Na(+)-K(+)-ATPase and Ca(2+)-Mg(2+)-ATPase were inhibited significantly, the membrane fluidity decreased and the membrane permeability increased. The fluorescence polarization and microviscosity in the low, middle and high dosage treatment groups were 0.0626 +/- 0.0051, 0.0685 +/- 0.0097, 0.0648 +/- 0.0086 and 0.3154 +/- 0.0298, 0.3510 +/- 0.0571, 0.3286 +/- 0.0504 respectively, compared with the control group (0.0481 +/- 0.0069 and 0.2338 +/- 0.0372) (P < 0.01).</p><p><b>CONCLUSION</b>DA has obvious effects on membrane function of rat glial cells and may cause further injury on the cells.</p>


Subject(s)
Animals , Rats , Cell Membrane , Cell Membrane Permeability , Cells, Cultured , Kainic Acid , Pharmacology , Membrane Fluidity , Neuroglia , Rats, Sprague-Dawley
14.
Chinese Journal of Industrial Hygiene and Occupational Diseases ; (12): 326-329, 2006.
Article in Chinese | WPRIM | ID: wpr-311494

ABSTRACT

<p><b>OBJECTIVE</b>To study the effects of deltamethrin (DM) on the permeability of mitochondrial membrane and the expression of cytochrome C in brain tissue of rats.</p><p><b>METHODS</b>Wistar rats were randomizedly divided into five groups (including four treated groups and one control group). In the treated groups, DM of 12.5 mg/kg was administered intraperitoneally once in rats and the rats were sacrificed 5, 24, 48 and 72 hours later while in the control group, the salad oil of 5 mg/kg was administered intraperitoneally once. The mitochondria in brain tissue of rats were extracted to measure the membrane permeability and the activity of cytochrome C oxidase as well as the expression of cytochrome C in cortex and hippocampus.</p><p><b>RESULTS</b>After the treatment the permeability of mitochondrial membrane was significantly increased in the treated groups compared with the control group. The expression of cytochrome C was increased in cortex and hippocampus CA1 and CA2 5 h, 24 h and 48 h groups and CA4 24 h group (0.57 +/- 0.04, 0.67 +/- 0.09, 0.58 +/- 0.04) and (0.81 +/- 0.18) (P < 0.05 or P < 0.01) while there was no significant difference in the expression of cytochrome C in cortex and hippocampus CA2 72 h group and CA3 and CA4 5 h, 48 h and 72 h groups between the treated groups and the control group (P > 0.05). The activity of cytochrome C oxidase was inhibited (P < 0.01).</p><p><b>CONCLUSION</b>Deltamethrin can significantly increase the permeability of mitochondrial membrane and the expression of cytochrome C in brain tissue of rats.</p>


Subject(s)
Animals , Male , Rats , Cerebral Cortex , Metabolism , Cytochromes c , Electron Transport Complex IV , Metabolism , Hippocampus , Metabolism , Mitochondrial Membranes , Metabolism , Nitriles , Pharmacology , Permeability , Pyrethrins , Pharmacology , Random Allocation , Rats, Wistar
15.
Chinese Journal of Biotechnology ; (12): 789-793, 2005.
Article in Chinese | WPRIM | ID: wpr-237073

ABSTRACT

Based on the technology of protein separation with immobilized metal ion affinity chromatography (IMAC), a method for oriented immobilization of papain has been selected. Papain was successfully immobilized on magnetic agarose carriers. Cu2+ with iminodiacetate (IDA) was used as the chelating ligand to be correlated with the histidine on papain (His-81). The optimum immobilization conditions of enzyme were as flows: Cu2+ 0.15 x 10(-2) mol/g carriers, time was 4h, enzyme load was 30 mg/g carrier, pH was 7.0, respectively. The pH and temperature were 8.0 and 70 degrees C for immobilized enzyme. The recovery activity of immobilized enzyme was retained 68.4%. The carrier could be recovered from the spent immobilized enzyme, to be reused. After 5 times, the reimmobilization of papain on the regenerated matrix was 79.71% effective with the retention of maximum enzyme activity. The cost of carriers used for industrial applications is very important. The regenerability of carriers is therefore, relevant. The mild conditions used for immobilization, the high recovery of immobilized preparations, the stability and the regenerability of the matrix are the main features of the method reported here. All above indicate this method can be applicable and promising in enzyme immobilization field.


Subject(s)
Catalysis , Chelating Agents , Chemistry , Chromatography, Affinity , Methods , Copper , Chemistry , Enzyme Stability , Enzymes, Immobilized , Chemistry , Hydrogen-Ion Concentration , Imino Acids , Chemistry , Metals , Papain , Chemistry , Temperature
16.
Journal of Experimental Hematology ; (6): 381-382, 2004.
Article in Chinese | WPRIM | ID: wpr-352059

ABSTRACT

The study was to observe the effect of rhG-CSF (lishengsu) in treating leukopenia caused by radiotherapy and chemotherapy in patients with breast cancer. 100 cases of breast cancer received modified radical mastectomy were randomized into two groups with the same treatment of one cycle chemotherapy using the protocol of CAF at two weeks after the operations and then radiotherapy. The patients in treated group received rhG-CSF 75 micro g per day s.c. for 5 - 7 days constantly, and additional 3 - 5 days according to leukopenia during radiotherapy. The patients in control group did not receive rhG-CSF during the chemo- and radio-therapy. The results shows that nadir of WBC and neutrophil counts in the treated group was higher than that in control significantly. In conclusion, effect of lishengsu on leucopenia in process of chemotherapy and radiotherapy shows definite therapeutic effect, the side effects are not remarkable.


Subject(s)
Adult , Female , Humans , Middle Aged , Breast Neoplasms , Therapeutics , Combined Modality Therapy , Granulocyte Colony-Stimulating Factor , Therapeutic Uses , Leukopenia , Drug Therapy , Recombinant Proteins
17.
Chinese Journal of Biotechnology ; (12): 287-290, 2004.
Article in Chinese | WPRIM | ID: wpr-259108

ABSTRACT

Numerous approaches and materials have been explored for preparation of immobilization enzymes because they have considerable advantages over enzyme in bulk solution. Among present enzyme carriers inorganic materials have lower cost and more stability than organic materials. But the present inorganic materials contain less active sites and have to undergo some complicated processes before binding enzyme. So it is necessary to explore a new kind of inorganic enzyme carrier. Papain, a well characterized thiol protease kinetically and structurally, is a suitable model to compare the efficiency of various immobilization procedures. SiO2 particles containing amine groups were synthesized by synchronous hydrolysis of tetraethylorthosilicate (TEOS) and N-(2-aminoethyl)-3-aminopropyltrimethoxysilane (AEAPS) in W/O Microemulsion of Triton X-100/cyclohexane/ammonium hydroxide system. The obtained particles have a diameter range from 0.3 microm to 0.5 microm, and their average size is 0.37 microm and can be controlled by adjusting water to surfactant molar ratio and water to TEOS and AEAPS molar ratio. Compared with traditional porous silica beads, these particles contain much more amine groups and their amine group content can be easily changed in the process of synthesis. Papain was immobilized on the particles which were treated by glutaraldehyde with covalent method. The optimum immobilization conditions of enzyme were as follows: enzyme load was 15 mg/g carrier, pH was 6.5. The pH and temperature optima were 6.5 and 70 degrees C for immobilized enzyme. All above indicate this kind of particle can be a good enzyme immobilization carrier.


Subject(s)
Amino Acids , Chemistry , Enzyme Stability , Enzymes, Immobilized , Chemistry , Metabolism , Papain , Chemistry , Metabolism , Particle Size , Silicon Dioxide , Chemistry
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