Evaluation method of medicine nature of individual ginsenosides / 中国药理学通报

.Aim To establish the evaluation methods of the med¬icine nature of ginsenosides by eorrelation analysis between med¬icine nature and ginsenoside contents in five Panax herbs with different medicine nature and the measurement of their effects on Na + /K + -ATPase activities.Methods 'Hie contents of ginsen¬osides in Sanqi, red ginseng , ginseng , American ginseng and ginseng leaves in the existing literature were eolleeted.and the medicinal nature was assigned by vector methods.rI1ie medicine nature of ginsenosides and the contribution of ginsenoside to the medicine nature were evaluated through bivariate correlation a- nalysis and grey eorrelation degree respectively.'Hie effects of ginsenosides on the Na+ /K +-ATPase activities of L02 eells and in pig eerebral cortex were measured to evaluate the medicine na¬ture of ginsenosides.Results Correlation results indicated that the order of correlation coefficients of ginsenosides to the warm and hot medicine nature was Rf > R1 > Rg3 > Rg2 > Rbl > Ro, while the order of correlation coefficients of ginsenosides to the eool and eold medicine nature was Rb2 > Re > Rd > Fll.Grey eorrelation degree analysis showed that the contribution of ginsen¬oside to the medicine nature was F11 > Re > Kg2 > Rd > Rb2 >Rbl > Rgl > Rc > Rg3 > R1 > Rf > Ro.The effects of ginsen- osides on Na +/K +-ATPase activities showed that the substance basis of the warm medicine nature was ginsenoside Rbl, Rb3, Rc, Rf, Rg2, Rgl, Rhl, Rg3, R1 and Ro, which increased the activity of Na + /K + -ATPase.While the cold and cool medi- cine nature were ginsenoside Rh2, Rd, Rh2, Re and oleanolic acid, which inhibited the activity of Na+/K

Discovery of a novel coronavirus associated with the recent pneumonia outbreak in humans and its potential bat origin

Since the SARS outbreak 18 years ago, a large number of severe acute respiratory syndrome related coronaviruses (SARSr-CoV) have been discovered in their natural reservoir host, bats1-4. Previous studies indicated that some of those bat SARSr-CoVs have the potential to infect humans5-7. Here we report the identification and characterization of a novel coronavirus (nCoV-2019) which caused an epidemic of acute respiratory syndrome in humans, in Wuhan, China. The epidemic, started from December 12th, 2019, has caused 198 laboratory confirmed infections with three fatal cases by January 20th, 2020. Full-length genome sequences were obtained from five patients at the early stage of the outbreak. They are almost identical to each other and share 79.5% sequence identify to SARS-CoV. Furthermore, it was found that nCoV-2019 is 96% identical at the whole genome level to a bat coronavirus. The pairwise protein sequence analysis of seven conserved non-structural proteins show that this virus belongs to the species of SARSr-CoV. The nCoV-2019 virus was then isolated from the bronchoalveolar lavage fluid of a critically ill patient, which can be neutralized by sera from several patients. Importantly, we have confirmed that this novel CoV uses the same cell entry receptor, ACE2, as SARS-CoV.

Substance basis of warm nature of Poria cocos / 中草药·英文版

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.

Analysis of Correlation between Gene Polymorphisms of CRHBP in Yunnan Han Population and Schizophrenia and Aggressive Behavior / 法医学杂志

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.

Observation on therapeutic efficacy of tuina plus cupping for chronic fatigue syndrome / 针灸推拿医学（英文版）

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.

Identification of a novel coronavirus causing severe pneumonia in human: a descriptive study / 中华医学杂志(英文版)

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.

Identification of a novel coronavirus causing severe pneumonia in human: a descriptive study / 中华医学杂志(英文版)

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.

Effects of hesperidin on the proliferation and expression of cyclin D in human pterygium fibroblasts / 国际眼科杂志(Guoji Yanke Zazhi)

@#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.

Effects of HDAC Inhibitor Scriptaid on IM9 Cell Line and Its Mechanism / 中国实验血液学杂志

<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>

Metabolic and excretory process of Citri Reticulatae Pericarpium Viride and Citri Reticulatae Pericarpium in rats' urine and feces analyzed by RRLC-QqQ-MSn / 中国中药杂志

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.

Highly Sensitive and Specific Detection of SRY Gene for Non-invasive Prenatal Diagnosis / 分析化学

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.

Characterization of Highly Pathogenic Avian Influenza H5N1 Viruses Isolated from Domestic Poultry in China / 生物医学与环境科学（英文）

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.

Highly Sensitive and Specific Detection of SRY Gene for Non-invasive Prenatal Diagnosis / 分析化学

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.

Characterization of 23S rRNA gene mutation in primary and secondary clarithromycin-resistant Helicobacter pylori strains from East China.

BACKGROUND/AIMS: Clarithromycin is an effective antibiotic for treating Helicobacter pylori; however, the development clarithromycin- resistance by multiple strains prevents the eradication of Helicobacter pylori. We aimed to characterize mutations in the 23S rRNA gene of primary clarithromycin-sensitive, primary clarithromycin-resistant and secondary clarithromycin-resistant Helicobacter pylori strains that were collected in East China and elucidate the mechanisms of clarithromycin resistance. MATERIALS AND METHODS: The disk diffusion test and E-test method were used to determine the clarithromycin susceptibility of clinical Helicobacter pylori strains. The 23S rRNA gene fragments were amplified by polymerase chain reaction from 18 primary clarithromycin- resistant strains, 15 primary sensitive strains and 8 secondary clarithromycin-resistant strains. Polymerase chain reaction-products were sequenced to determine mutations of the 23S rRNA gene. RESULTS: We found an A2143G (8 strains) mutation in primary clarithromycin-resistant strains, an A2143T (5 strains) mutation in secondary clarithromycin-resistant strains; but no mutations were found in position 2143 of sensitive strains. A T2182C mutation in primary clarithromycin-sensitive, primary clarithromycinresistant and secondary clarithromycin-resistant strains was found with a prevalence of 86.7% (13 strains), 72.2% (13 strains) or 87.5% (7 strains), respectively. In addition, we found a G2254T (8 strains) and a G2172T (7 strains) mutation in secondary clarithromycin- resistant strains. These point mutations were absent in primary clarithromycin-resistant and -sensitive strains. CONCLUSION: The gene mutation in position 2143 was associated with resistance to clarithromycin, but the mutation was different between primary and secondary clarithromycin-resistant strains. The T2182C mutation was not associated with clarithromycin resistance. Two new hotspot mutations: G2254T and G2172T, in 23S rRNA were discovered in secondary clarithromycin-resistant strains.

Analysis of severe complications after transcatheter arterial chemoembolization for primary hepatocellular carcinoma / 中华肿瘤杂志

<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>

Effects of domoic acid on membrane function of primary cultured rat glial cells / 中华劳动卫生职业病杂志

<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>

Tetrandrine-induced apoptosis in rat primary hepatocytes is initiated from mitochondria: caspases and endonuclease G (Endo G) pathway.

Tetrandrine, a bisbenylisoquinoline alkaloid isolated from the dried root of Stephenia tetrandra (S Moore), possesses a remarkable pharmacological profile. However, the mechanisms of tetrandrine hepatotoxicity remain to be elucidated. In this study, we first proved apoptosis and mitochondrial dysfunction induced by tetrandrine in Sprague-Dawley rat liver in vivo. By further assuming apoptosis as an important mechanism in tetrandrine-induced hepatotoxicity, we focused on mitochondria-initiated apoptosis in primary hepatocytes isolated from Sprague-Dawley male rats. Tetrandrine treatment led to significant release of cytochrome c and downregulation of Bcl-X(L) accompanied by caspase 3 activation, and ultimately, DNA fragmentation. Caspase 3 activation was markedly inhibited by cyclosporin A (CsA) and Ac-DEVD-CHO. Furthermore, Endo G, a caspase-independent apoptotic protein, was detected for its expression and DNase activity. CsA blocked the release both of Endo G and cytochrome c significantly. Additionally, the generation of reactive oxygen species (ROS) increased in a time-dependent manner corresponding with a fall in intracellular GSH content after 10 microM tetrandrine treatment in 4h. Tetrandrine also induced mitochondrial dysfunction indicated by transition of mitochondrial transmembrane potential and decrease of intracellular ATP level. The findings indicated that the caspase-dependent mitochondrial apoptosis pathway was primarily involved in tetrandrine-induced apoptosis in rat primary hepatocytes. In addition, a caspase-independent pathway indicated by Endo G also contributed to apoptosis caused by tetrandrine. Meanwhile, ROS was proved an important inducer in this apoptosis process.

Effects of deltamethrin on permeability of mitochondrial membrane and expression of cytochrome C in brain tissue of rats / 中华劳动卫生职业病杂志

<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>

Oriented immobilization of papain on metal chelating carriers / 生物工程学报

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.

Immobilization and stabilization of papain on SiO2 particles containing amine groups / 生物工程学报

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.