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.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
7.Analysis on regulating deqi in the ancient works of the late dynasties.
Jie HAO ; Jiang ZHU ; Peng ZHANG ; Dandan QI ; Nijuan HU ; Chi LIN ; Pei WANG ; Minyi ZHAO ; Shangqing HU ; Guiwen WU ; Siyuan XIN
Chinese Acupuncture & Moxibustion 2015;35(11):1173-1176
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
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history
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methods
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Books
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history
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China
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History, Ancient
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Humans
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Medicine in Literature
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Qi
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history
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Sensation
8.Construction of the subtractive cDNA library of injured adult and fetal rabbit skins.
Bo ZHANG ; Da-wei LIU ; Zheng-guo WANG ; Pei-fang ZHU ; Ji-hong ZHOU ; Jian-xin JIANG
Chinese Journal of Traumatology 2004;7(1):13-17
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
9.Acupuncture inhibiting responses of spinal dorsal dorsal horn neurons induced by noxious dilation rectum and colon.
Pei-jing RONG ; Bing ZHU ; Qi-fu HUANG ; Xin-yan GAO ; Hui BEN ; Yan-hua LI
Chinese Acupuncture & Moxibustion 2005;25(9):645-650
OBJECTIVETo study on mechanisms of acupuncture in relieving visceral pain.
METHODSIn SD rats CRD was used as noxious visceral stimuli. Activities of spinal dorsal horn wide dynamic (WDR) neurons of L1-L13 were recorded by extracellular microelectrode technique. Acupuncture was given at ipsi-lateral and contra-lateral Zusanli (ST 36) of the same segmental innervation of rectum and colon.
RESULTSVisceral noxious afferent could significantly activate spinal dorsal horn convergent neurons, and mechanical stimulation of contra-lateral body surface and hand acupuncture at Zusanli (ST 36) could inhibit this noxious response. When the spinal cord was acutely blocked, the inhibiting CRD effect of needling CRD effect of needling contra-lateral Zusanli (ST 36) completely disappeared.
CONCLUSIONAcupuncture and visceral noxious afferent signals converge and interact each other in spinal level, and acupuncture at acupoint can inhibit the spinal dorsal horn neuron respon se activated by visceral noxious afferent and this action needs the participation of the center above the spinal cord.
Animals ; Colon ; innervation ; Nociceptors ; Posterior Horn Cells ; Rats, Sprague-Dawley ; Rectum ; Spinal Cord
10.Study on trauma of tympanic membrane and ossicular chain of guinea pigs exposed to experimental blast underpressure
Chao-Jun LI ; Zhao-Hua LIU ; Pei-Fang ZHU ; Zheng-Guo WANG ; Cheng YANG ; Hai-Bin CHEN ; Xin NING ; Ji-Hong ZHOU ;
Chinese Journal of Trauma 2003;0(08):-
Objective To probe into the characteristics of traumatic effect of blast underpressure (BUP) on tympanic membrane and ossicular chain of guinea pigs exposed to blast underpressure. Methods The guinea pigs were exposed to experimental BUP generated by the underpressure generator and their tympanic membranes and auditory ossicules were observed with stereo-microscope.Results Most of the tympanic membranes in 72 guinea pigs (141 ears) presented with traumatic changes,such as engorgement,haemorrhage,epidermic desquamation and perforation,combined with fracture of ossicles in some guinea pigs.The traumatic severity of the tympanic membranes and auditory ossicules was related to the peak and the declinating duration of BUP.The minimum peak of the underpressure inducing perfo- ration was between-22.4kPa and-23.9kPa and the minimum peak inducing perforation of all the tym- panic membranes was between-83.1kPa and-87.2kPa.Conclusion The tympanic membrane in guinea pigs exposed to BUP changed traumatically,such as engorgement,haemorrhage,epidermic des- quamation,perforation and fracture of ossicles.The traumatic severity of the tympanic membranes and au- ditory ossicules is related to the peak and declinating duration of BUP.