2.Construction, expression and preliminary pharmacokinetic analysis of IL-1ra mutants.
Yu-Xin WANG ; Zhi-Xin YANG ; Heng-Qi ZHU ; Xiao-Wei ZHOU ; Pei-Tang HUANG
Chinese Journal of Biotechnology 2006;22(3):472-476
Interleukin-1 receptor antagonist (IL-1ra), a member of IL-1 family, is a naturally occurring IL-1 inhibitor as "receptor antagonist", which blocks biological responses mediated by IL-1. Recombinant human IL-1ra (rhIL-1ra, Kineret) was introduced in clinical trials involving patients with RA. Between 2001 to approximately 2002, rhIL-1 ra was approved by the US Food and Drug Administration and the European Agency for the Evaluation of Medicine Procedure. Unfortunately, 10,000 to 100,000-fold excess amounts of IL-1ra are needed to relieve disease because minimal IL-1 can induce complete biological responses, and the dosage of 100 to approximately 150mg/day in a RA patient is so big that it greatly influence patients' physical, psychological and economical situation. In this study, IL-1ra mutants were established by site-specific mutagenesis to improve its stability. The sites of mutagenesis included R6 K7-AA,R93 K94-AA and K97 R98-AA. IL-1ra and its mutants were expressed in E. coli BL21 (DE3) using pTIG-Trx expressing system with the induction of IPTG. The recombinant proteins were purified by Ni2+ chelate chromatography and Sephadex G75 gel filtration chromatography. The activity of mutants is as high as IL-1ra. We characterized the pharmacokinetic profile of IL-1ra and its mutants. The third mutant's half life is 2.26 times than wt IL-1ra. The study has provided some approaches and experience for further research to improve the metabolism stability of IL-1ra.
Animals
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Escherichia coli
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genetics
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metabolism
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Female
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Humans
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Interleukin 1 Receptor Antagonist Protein
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biosynthesis
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genetics
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pharmacokinetics
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Mutagenesis, Site-Directed
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methods
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Mutant Proteins
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biosynthesis
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pharmacokinetics
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Recombinant Proteins
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biosynthesis
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genetics
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pharmacokinetics
3.Preparation and evaluation of novel solid chlorine dioxide-based disinfectant powder in single-pack.
Min ZHU ; Li-Shi ZHANG ; Xiao-Fang PEI ; Xin XU
Biomedical and Environmental Sciences 2008;21(2):157-162
OBJECTIVETo prepare and evaluate novel chlorine dioxide-based disinfectant powder in single-pack that is more convenient for use and transportation.
METHODSOrthogonal experiment was performed to determine the recipe of the disinfectant powder. Stability test, suspension quantitative bactericidal test, simulation field trial, and animal toxicity test were carried out to observe its bactericidal and toxicological effects.
RESULTSThe orthogonal experiment showed that the type of water solution had no effect on the disinfectant powder and the best ratio of sodium chlorite to solid acid was 1:3. Ten grams of the disinfectant powder was fully dissolved in 20 mL water for 2 min, and diluted to 500 mL in water. After 5-10 min, the concentration of chlorine dioxide (ClO2) solution was 266 mg/L to 276 mg/L. After stored at 54 degrees C for 14 d, the average concentration of ClO2 was decreased by 5.03%. Suspension quantitative bactericidal test showed that the average killing logarithm (KL) value for both Staphylococcus aureus and Escherichia coli in 100 mg/L ClO2 solution for 2 min was over 5.00. in simulation field trial, the average descending KL value for Escherichia coli in the solution containing 100 mg/L ClO2 for 5 min was over 3.00. The mouse acute LD50 in the solution 5 times exceeded 5000 mg/kg. The disinfectant powder was not toxic and irritative to rabbit skin and had no mutagenic effect on mouse marrow polychromatic erythrocytes (PCE).
CONCLUSIONThe stability and bactericidal efficacy of solid chlorine dioxide-based disinfectant powder in single-pack are good. The solution containing 100 mg/L ClO2 can kill vegetative forms of bacteria. The concentration of ClO2 on the disinfecting surface of objects is 100 mg/L. The disinfectant powder is not toxic and irritative.
Chlorine Compounds ; pharmacology ; Disinfectants ; pharmacology ; Escherichia coli ; drug effects ; Oxides ; pharmacology ; Staphylococcus aureus ; drug effects
4.The correlative study on the occurrence of negative emotion of advanced cancer patients and the emotional changes of caregivers
Lin ZHU ; Qing-Xin WANG ; Yi PEI
Chinese Journal of Modern Nursing 2010;16(20):2360-2362
Objective To explore whether the change of emotion of patients with malignant tumors affect their family member and other people around. Methods 62 cases of hospitalized cancer patients and their family member were respectively investigated by using self-rating anxiety scale (SAS) at the same time.According to the result of SAS, patients and their family member were divided into anxious group and nonanxious group. In incidence of negative emotion and abreaction status were compared between the patients group and the family member group. Results 88.71% patients occurred anxiety symptom and 96. 36% patients vented their negative emotion. 85.48% patients' family members occurred anxiety and 94.34% vented their negative emotion. Incidence of anxiety and abreaction status was no significant deference between patients and their family member. Conclusions The negative emotion of advanced cancer patients affected their family member and other people around. Those affected people vented their negative emotion to others. Therefore,psychological nursing should be taken in early stage both for patients and their family member.
5.Establishment and neurochemical studies of animal models with anterior transposition of the ulnar nerve
Ya-Zhong ZHU ; Sui-Pei WU ; Li-Xin YU ; Rong ZHOU ; Xin-Hong ZHU ; De-Sheng DONG ; Shu-Cai LING ; Chun-Sheng WANG
Chinese Journal of Neuromedicine 2010;9(3):258-261
Objective To establish animal models with anterior transposition of the ulnar nerve and evaluate the safety of anterior transposition of the ulnar nerve at molecular level.Methods Location of the ulnar nerve of elbow in 5 rats were found similar to human being by anatomy.Twenty healthy adult SD rats,weighting about 250 g,were performed the anterior transposition of the ulnar nerve in the right forelimbs and the left forelimbs was considered as control group.The bilateral flexor carpi ulnaris muscles were weighed and the slice of cervical spinal cord(C_6-T_1)level were prepared 1 month after the operation.Nissl staining,NADPH-d histochemical staining,IB4 staining and ChAT-immunohistochemical staining were employed to observe the spinal cord(C_6-T_1)level at molecular level;electron microscope was used to observe the ultrastructure of ChAT-positive neurons.Statistical analysis was paired T test.Results The flexor carpi ulnaris muscles in the model group(92.3±9.13mg)and control group(93.2±7.29 mg)were not significantly different(P>0.05).After anterior transposition of the ulnar nerve in rats,no significant differences in cell number and morphology in the cervical spinal cord(C_6-T_1)were found between the model group and the control group(P>0.05).No changes between the 2 groups were noted in the fine structure of anterior horn motor neurons and the expression of nenrotransmitters(P>0.05).Conclusion Anterior transposition of the ulnar nerve can be safely done in the animal models(rats).
6.Linkage disequilibrium analysis of -31, -511, -1470 single nucleotide polymorphisms in the promoter of IL-1 beta in Chongqing population.
Ai-qing WEN ; Jian-xin JIANG ; Pei-fang ZHU ; Kai FENG
Chinese Journal of Medical Genetics 2004;21(4):412-414
OBJECTIVETo investigate the frequencies of -31, -511 and -1470 single nucleotide polymorphisms (SNPs) in the promoter of IL-1 beta in Chongqing population and address the question whether they are in linkage disequilibrium (LD).
METHODSOne hundred and twelve healthy Chongqing people were enrolled in this study. Polymorphisms at -31 (C>T), -511 (T>C) and -1470 (G>C) of IL-1 beta were genotyped with the method of restriction fragment length polymorphism (RFLP). Haplotype frequencies were evaluated with Arlequine software. Two-point LD analyses were done with the software TransposerV1-0. The P values were determined by Pearson chi square test analysis.
RESULTSAllelic frequencies of IL-1 beta -31, -511 and -1470 were 45.33%, 50.00% and 41.67%, respectively. The dominant haplotypes comprising the three loci were T-C-G (44.1%), C-T-C(40.3%) and C-T-G(8.8%), LD analyses revealed that none of the LD parameters(delta value) was 0. Meanwhile, chi square test showed P<0.005.
CONCLUSION-31, -511 and -1470 loci in the promoter region of IL-1 beta are in strong linkage disequilibrium. And this study provides a basis for searching disease-related IL-1 beta haplotye.
Adult ; China ; Female ; Gene Frequency ; Genotype ; Haplotypes ; Humans ; Interleukin-1beta ; genetics ; Linkage Disequilibrium ; Male ; Middle Aged ; Polymorphism, Restriction Fragment Length ; Polymorphism, Single Nucleotide ; Promoter Regions, Genetic ; genetics
7.Detection of cholesterol ester transfer protein in semen of infertile patients.
Yong-Xin LIU ; Pei-Yuan ZHU ; Jun-Jun WANG ; Jun MO ; Yu-Feng HUANG
National Journal of Andrology 2003;9(3):178-180
OBJECTIVESTo detect the cholesterol ester transfer protein (CETP) levels in semen of infertile patients and evaluate the correlation between CETP in semial plasma and infertility.
METHODSOne hundred and sixty-three infertile patients and fifteen fertile males were selected randomly. The routine examination of ejaculates was fulfilled by computer aided semen analysis (CASA). The CETP levels in all seminal plasma samples and fifty-five serum samples were detected by ELISA method.
RESULTSThe CETP levels in infertile patients and fertile males were (2.21 +/- 1.23) microgram/L and (1.40 +/- 0.45) microgram/L, respectively. There were no significant differences between the two groups(P > 0.05). And there were no significant differences of CETP levels in seminal plasma among groups of azoospermia(n = 29), oligoasthenozoospermia (n = 58), oligospermia(n = 15), asthenozoospermia(n = 44) and normozoospermia(n = 17) in the infertile patients(P > 0.05). The CETP in seminal plasma and serum were detected in 55 infertile patients, and there was no correlation between CETP levels in seminal plasma and serum using Spearman analysis(r = 0.009, P > 0.05). The mean CETP level in seminal plasma was almost 1/1,000 of that in serum.
CONCLUSIONSThe CETP level in seminal plasma is extremely low and has no relation with the changes of sperm density or motility. It may ensure the integrity of sperm membrane before the sperm enters into female genital tract.
Adult ; Carrier Proteins ; analysis ; blood ; Cholesterol Ester Transfer Proteins ; Glycoproteins ; Humans ; Infertility, Male ; metabolism ; Male ; Middle Aged ; Semen ; chemistry
8.Sepsis and membrane receptors.
Zhao-xia DUAN ; Pei-fang ZHU ; Jian-xin JIANG
Chinese Journal of Traumatology 2005;8(1):60-64
9.Analysis of polymorphisms in the promoter region of interleukin-1beta by restriction fragment length polymorphism-PCR.
Ai-Qing WEN ; Jun WANG ; Kai FENG ; Pei-Fang ZHU ; Jian-Xin JIANG
Chinese Journal of Traumatology 2004;7(5):271-274
OBJECTIVETo investigate the frequencies of -1470, -511 and -31 single nucleotide polymorphisms (SNPs) in the promoter of IL-1beta and its haplotype constitution in Chongqing population.
METHODSOne hundred and twelve healthy Chongqing people were enrolled in this study. Polymorphisms at -1470 (G to C), -511 (T to C) and -31 (C to T) of IL-1beta were genotyped with the method of restriction fragment length polymorphism (RFLP). Haplotype frequencies were analyzed by Arlequine software.
RESULTSFrequencies of IL-1beta -1470, -511 and -31 SNPs were 41.67%, 50% and 45.33%, respectively. Genotype frequencies of -1470 locus were 39.81%, 37.04% and 23.15% for G/G, G/C and C/C respectively. As for T-511C SNP, genotype frequencies of T/T, T/C and C/C were 29.91%, 40.18% and 29.91%, respectively. Genotyping results of C/C, C/T, and T/T of -31 locus were 35.51%, 38.32% and 26.71% respectively. Haplotype analysis found that there were mainly three haplotypes constituted by three SNPs, ie., G-T-C, C-T-C and G-C-T.
CONCLUSIONSPolymorphisms exist in the promoter of IL-1beta in Chongqing population. Three SNPs locate in the same haplotype block.
Asian Continental Ancestry Group ; genetics ; China ; Cohort Studies ; Female ; Genetics, Population ; Genotype ; Haplotypes ; Humans ; Interleukin-1 ; genetics ; Male ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; Polymorphism, Single-Stranded Conformational ; Promoter Regions, Genetic ; Sensitivity and Specificity
10.Comparison of 3 microsurgical approaches to the treatment of varicocele: report of 120 cases.
Qing SHI ; Hai ZHU ; Xin-Sheng WANG ; Pei-Tao WANG ; Si-Chuan HOU
National Journal of Andrology 2013;19(10):931-934
OBJECTIVETo compare the outcomes of the three microsurgical strategies, inguinal high ligation (IHL), retroperitoneal high ligation (RHL) and low ligation (LL) of internal spermatic veins, in the treatment of varicocele.
METHODSWe retrospectively analyzed 120 cases of varicocele, which were equally divided into groups I , II and III to be treated by IHL, RHL and LL of internal spermatic veins, respectively. We compared the operation times, post-operative complications, recurrence rates and results of pre- and post-operation semen analysis among the three groups.
RESULTSThe mean operation time was significantly longer in group III ( [55 +/- 6 ] min) than in I ([35 +/- 10] min) and II ([42 +/- 12] min) (P<0.05), while the rate of post-operative complications remarkably higher in group I (4 cases of hydrocele [10% ] and 3 cases of epididymitis [7.5%]) than in II (2 cases of hydrocele [5%] and 2 cases of epididymitis [5%]) and III (1 case of hydrocele [2.5%] and 1 case of epididymitis [2.5%]) (P<0.05). Six to 12 (mean 9) months of follow-up visit found 2 cases (5% ) of recurrence in group I, 1 case (2.5%) in group II and none in group III, with no statistically significant difference among the three groups (P>0.05). At 12 months after surgery, group III showed significantly higher sperm concentration, grade a +b sperm and the sperm motility than the other two (P<0.05), but no significant differences were observed in these parameters among the three groups preoperatively.
CONCLUSIONAs a microsurgical approach to the treatment of varicocele, LL is better than IHL and RHL of internal spermatic veins in improving the seminal parameters of the patients.
Humans ; Ligation ; Male ; Microsurgery ; adverse effects ; methods ; Retrospective Studies ; Treatment Outcome ; Urologic Surgical Procedures, Male ; adverse effects ; methods ; Varicocele ; surgery