Animals ; Biological Warfare ; Burns ; pathology ; Burns, Chemical ; pathology ; Chemical Warfare ; China ; Humans ; Military Medicine ; Radiation Injuries ; pathology ; Wounds, Gunshot ; pathology

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 ; Escherichia coli ; genetics ; metabolism ; Female ; Humans ; Interleukin 1 Receptor Antagonist Protein ; biosynthesis ; genetics ; pharmacokinetics ; Mutagenesis, Site-Directed ; methods ; Mutant Proteins ; biosynthesis ; pharmacokinetics ; Recombinant Proteins ; biosynthesis ; genetics ; pharmacokinetics

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

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).

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

From the time of Neijing (Internal Classic) and Nanjing (Classic of Medical Problems) till the end of Qing dynasty, the medical scholars of each dynasty had taken the recognition of deqi as the subject in the stud- ies. Through the historical analysis, the content of regulating deqi was further understood in relevant ancient liter- ature. By checking the ancient works of acupuncture in each dynasty till the end of Qing dynasty, in reference to the evidences in over 10 works, such as Zhenjiu Dacheng (Great Compendium of Acupuncture and Moxibustion) , Zhenjing Zhinan (Instruction of Acupuncture Canon), Zhenjiu Daquan, and in association with the academic views of acupuncture masters in modern time, it was discovered that the medical scholars after the time of Neijing and Nanjing had enriched the understandings of deqi, such as the connection of cold and heat reaction, radiation to the affected site, reinforcing and reducing purpose in qi regulation. The methods of deqi regulations had, been explored till the end of Qing dynasty since the time of Neijing and Nanjing and the understandings of it were vari- ous among scholars.
Acupuncture Therapy ; history ; methods ; Books ; history ; China ; History, Ancient ; Humans ; Medicine in Literature ; Qi ; history ; Sensation

OBJECTIVEEarly gestational mammalian fetuses possess the amazing ability to heal cutaneous wounds in a scarless fashion. Over the past years, scientists have been working to decipher the mechanisms underlying this regenerative repair. The remarkable phenotypic differences between fetal and adult healings behooves us to learn their characteristics in genetics, which represents potentially important mechanisms involved in wound repair observed in fetal versus adult tissues. In this sense, it is reasonable to construct subtractive cDNA library for future research.

METHODSMiddle laparotomy and hysterotomy were performed on pregnant rabbits at 20-day gestation to expose the fetal back, and a longitudinal incision through the skin was made on the back of the fetus. The traumatized fetal skin was harvested 12 hours post-operation, the fetus control and traumatized adult skin specimens were taken at the same time. dscDNA was synthesized from total RNA of skin samples with SMART technology. Taking one of the three samples as Tester respectively and the other two as Drivers, we obtained 1 forward and 2 reverse hybridization products. After being amplified with selective polymerase chain reaction, the products were inserted into a vector, and then transferred into E. coli HB101. The colonies were screened afterwards.

RESULTSThe wounded fetuses were alive for a long time even after birth. Every determinant step, such as RNA isolation, cDNA synthesis, Rsa I digestion, adaptor ligation and hybridization, was well-operated. Subtractive efficiency identification demonstrated that the suppression subtractive hybridization (SSH) was successful. Insertion into vector and transferring to E. coli were satisfactory.

CONCLUSIONSInstead of classic SSH, an improved SSH with 2 Drivers was applied for the experiment. Results confirmed that the improved program was reasonable and correct in both theory and practice. The subtractive cDNA library we have obtained is going to be used for future researches to reveal scarless healing related gene(s) and its (their) expression.

Animals ; Cicatrix ; genetics ; DNA, Complementary ; Disease Models, Animal ; Female ; Fetus ; Gene Library ; Polymerase Chain Reaction ; Pregnancy ; Pregnancy, Animal ; RNA, Messenger ; analysis ; Rabbits ; Sensitivity and Specificity ; Skin ; injuries ; Wound Healing ; genetics ; physiology

Acute-Phase Proteins ; physiology ; Carrier Proteins ; physiology ; Humans ; Membrane Glycoproteins ; physiology ; Receptors, Cell Surface ; physiology ; Receptors, Immunologic ; physiology ; Receptors, Scavenger ; Sepsis ; metabolism ; physiopathology ; Signal Transduction ; Toll-Like Receptors

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

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