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

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

Acupuncture-moxibustion theory believes that needling sensation is a crucial factor in influencing acupuncture efficacy. Judgment of occurrence of needling qi (qualitative measurement) and the intensity of needling qi (quantitative measurement) is the key section in studying the relationship between needling qi and efficacy. According to the relevant literatures, the judgment of the occurrence of needling qi mostly depended on the needling sensations, while a small amount of researches mentioned self-determined threshold or range of needling qi; the quantity of needling qi was measured by factor analysis, Massachusetts General Hospital Acupuncture Sensation Scale (MASS) based on exponential smoothing method, weighted average, addition of needling sensation score, etc. This article holds that the needling qi should be divided into two parts: overall needling qi and needling sensation components, the former referring to the occurrence and intensity of needling qi, and the latter for judging the different sensations and intensity. Currently, the studies on needling qi and efficacy should begin with single treatment at single point and multiple treatments at multiple points, to generally estimate the quality and quantity of needling qi, for seeking a qualitative and quantitative measurement in accordance with both acupuncture-moxibustion theory and clinical practice.

ObjectiveTo explore the relation between the uterine position and acupoint effect by analyzing the data of a clinical trial of electroacupuncture in treating primary dysmenorrhea.MethodThe uterine position was detected by ultrasonic examination;Visual Analogue Scale (VAS) was used to evaluate the pain degree before and after intervention; Retrospective Symptom Scale (RSS) was adopted to determine the improvement of symptoms.ResultThere were no significant differences in comparing the VAS score, real-time effect and post-treatment effect, and effective rate among different uterine positions (P>0.05). Electroacupuncture at Sanyinjiao (SP 6) can produce a real-time effect in releasing abdominal pain and relevant symptoms of dysmenorrhea in patients with anteversion of uterus, a less significant effect was shown in patients with retroposition of uterus, while no effect was shown in patients with uterus at middle position.ConclusionElectroacupuncture at Sanyinjiao possibly has a specific effect in releasing abdominal pain and relevant symptoms of dysmenorrhea at anteversion of uterus, and the uterine position may be related to the corresponding meridians and Zang-fu organs. The current statistical result indicates that there is no relation between the uterine position and the effect of Sanyinjiao, but this conclusion still needs proving by prospective randomized controlled clinicaltrials.

Objective To explore the effect of vitamin D hormone(VDH)on autophagy and apoptosis in hippocampus of rats with traumatic brain injury(TBI). Methods A total of 45 male Sprague-Dawley rats were divided into control group(n=15),model group(n=15)and VDH group(n=15).The TBI model was established in the model group and VDH group.VDH group was injected with VDH 1 μg/kg 30 minutes,24 hours and 48 hours after modeling,respectively.The expression of microtubule as-sociated protein 1 light chain 3(LC3)and p62 was detected with Western blotting,and the number of apoptotic cells were detected with TUNEL three days after modeling.All groups were tested with Morris water maze on the fifth,sixth and seventh days. Results The number of TUNEL positive cells in hippocampus was more in the model group than in the control group (P<0.05),and was lower in VDH group than in the model group(P<0.05),as well as the expression of LC3II/LC3I and p62(P<0.05).The escape latency was longer in the model group than in the control group(P<0.05),and was shorter in VDH group than in the model group (P<0.05). There was no significant difference in the swimming velocity on the fifth,sixth,and seventh days among three groups(F=0.530,P>0.05). Conclusion VDH has potential neuroprotective effects on TBI,which might be associated with its anti-apoptosis effect on the expression of LC3 and p62 proteins in hippocampus after TBI.

Objective: To explore the relationship between serum bilirubin level and the severity of coronary artery disease (CAD). Methods: A total of 568 patients treated in our hospital from 2014-07 to 2016-03 were studied. Based on coronary angiography (CAG), the patients were divided into 2 groups: CAD group, n=438 and Non-CAD group, n=130. CAD group had 166 patients with single vessel disease, 150 with double vessel disease and 122 with triple vessel disease; taking Gensini score 34 as the border, there were 219 patients with low Gensini score and 219 with high Gensini score. Taking total bilirubin (TBIL) level>17.1 μmol/L as the border, the patients were divided into another set of 2 groups: Normal TBIL group, n=426 and High TBIL group, n=142. Clinical data, biochemical parameters, blood routine examination, color doppler ultrasound and CAG results were recorded. Blood levels of TBIL, direct bilirubin (DBIL) and indirect bilirubin (IBIL) were compared among different groups. Results: ①Compared with Non-CAD group, CAD group had decreased serum levels of TBIL, DBIL and IBIL, P<0.05, the detective rate of hyperbilirubinemia had decreasing trend; while detective rate of CAD was similar between Normal TBIL group and High TBIL group, P>0.05. ②In CAD group, the patients with more coronary branch lesions and the higher Gensini scores had the lower serum levels of TBIL, DBIL and IBIL, P<0.05. Pearson correlation analysis showed that TBIL, DBIL and IBIL were positively related to LVEF (r=0.390, r=0.283 and r=0.426), all P<0.05. ③Multi-linear regression analysis indicated that DBIL, IBIL level was the independent risk factor for CAD occurrence and having negative correlation to CAD (B=-3.766, B=-2.705), both P<0.05. ④TBIL, DBIL and IBIL levels were negatively related to hs-CRP in CAD patients (r=-0.617, r=-0.473 and r=-0.580),allP<0.05. Conclusion: Serum bilirubin level was negatively correlated to the degree of the cornary stenosis, which is independent of its forms, low bilirubin might be an independent risk factor of CAD occurrence.