1.The Research on Expression and Purification of a Chimeric Anti-p185 Antibody
Jing-Juan QIAO ; Hai-Bo LIU ; Juan-Juan ZHU ; Lian-Sheng CHENG ; Jing LIU ;
China Biotechnology 2006;0(09):-
The Her-2 proto-oncogene encodes a 185kDa transmenbrane glycoprotein p185 which has intrinsic tyrosine kinase activity. It is overexpressed in several malignant human tumors like breast cancer. A chimeric antibody by assembling a single-chain Fv antibody and a human IgG1 Fc fragment was constructed. This chimeric antibody reacts with tumor surface antigen p185c-erbB-2 specifically. In order to put the antibody into clinical application, two steps purification method was used to attain the antibody’s purity more than 95%. Both the lyophilized pharmaceutical formulations of the antibody were found. The formulations can keep the stability and activity of the antibody for at least one year. These results were the foundation of the chimeric antibody for cancer therapy.
2.Influence of Ganlu Xiaodu Dan on Expression of Toll-like Receptor 4 mRNA and Nuclear Factor ?B p65 in Rats with Damp-Heat Syndrome of Seasonal Febrile Disease
Fangping CHENG ; Jie ZHOU ; Juan CHEN ; Songlin LIU ; Guoqiang MEI
Journal of Guangzhou University of Traditional Chinese Medicine 2004;0(06):-
【Objective】To observe the dynamic interference of cellular endotoxin specific receptors of Toll-like receptor 4(TLR4) mRNA and nuclear factor ?B p65(NF-?B p65) in rats with damp-heat syndrome of seasonal febrile disease by Ganlu Xiaodu Dan(GXD),and to explore the therapeutic mechanism of prescriptions with the actions of clearing heat and resolving dampness.【Methods】Wistar rats were randomized into 3 groups: A(normal control),B(models established by feeding of high-fat diet and colon bacillus under the high-temperature and high-humidity condition),and C(rat models treated with GXD).Reverse transcription-polymerase chain reaction(RT-PCR) analysis was used to detect the TLR4 mRNA level in hepatic macrophages and immunohistochemical method was used to detect the activation of NF-?B p65.【Results】The expression of TLR4 mRNA and NF-?B p65 was increased gradually in group B 6,12 and 24 hours after infection with colon bacillus,and the differences were significant,indicating that the expression of TLR4 mRNA and NF-?B p65 increased with the prolongation of disease course.In group C,TLR4 mRNA expression was decreased in different time points after infection,the difference of TLR4 mRNA expression 12 and 24 hours after infection was significant as compared with group B and group A(P
3.Th cell differentiation in chronic stage of viral myocarditis in mice and interference of Qingxin-II Recipe
Zhiqing CHENG ; Hongfei LIU ; Juan ZHANG ; Hangping YAO
Journal of Integrative Medicine 2007;5(3):318-21
OBJECTIVE: To investigate part mechanisms of Th cell differentiation, and to observe the interference effect of Qingxin-II Recipe in the chronic stage of viral myocarditis (VMC), so as to provide some experimental evidences for illuminating the pathogenesis of VMC and treatment mechanisms of Qingxin-II Recipe. METHODS: According to 20%-40% death rate of experiment in advance, 100 BALB/c male mice (4 weeks old and weighing 12-15 g) were used. Twenty mice were randomly assigned to normal control group, and the other 80 mice were intraperitoneally injected with 0.1 ml normal saline containing coxsackie virus B3 at the 1st, 4th and 28th day (the virus densities were 1:2000, 1:1000 and 1:500 respectively) to induce chronic VMC. At the 42nd day, the surviving mice were randomly divided into untreated group and treatment group, with 20 mice in each group. Mice in the treatment group were orally administered with 0.2 ml Qingxin-II Recipe every day, while mice in the normal control group and the untreated group were administered with 0.2 ml normal saline. All the mice were sacrificed after 45 days, and the sera, heart and spleen cells were collected. Then the myocardial pathological changes were observed by using a light microscope, and the levels of IL-4, IL-10 and IFN-gamma in serum were detected by enzyme linked immunosorbent assay (ELISA). And the Th cell differentiation was observed by flow cytometry. RESULTS: No obvious myocardial pathological changes were observed in mice of the normal control group. Myocardial pathological changes in the treatment group were slighter than those in the untreated group. The difference of serum IL-10 level between the normal control group and the untreated group showed no significance (P>0.05), and the levels of IFN-gamma and IL-4 of the untreated group were higher than those of the normal control group (P<0.05 or P<0.01). There was no statistical difference in IL-10 level between the treatment group and the untreated group (P>0.05), while the serum levels of IL-4 and IFN-gamma of the treatment group were lower than those of the untreated group (P<0.05 or P<0.01). There was no significant difference of the Th1 cell responder between the treatment group and the untreated group (P>0.05), while the Th2 cell responder was inhibited significantly in the treatment group (P<0.05). CONCLUSION: Qingxin-II Recipe can restore the balance of Thl and Th2 cells through inhibiting the reaction of Th2.
4.Clinical analysis of Compound Anisodine Injection combined with Iodized Lecithin Tablets in treatment of central serous chorioretinopathy
Xuan LIU ; Juan DU ; Cheng CHENG ; Ni WANG ; Mengrui ZHONG ; Jiangkun WU ; Yin HAO
Drug Evaluation Research 2017;40(2):249-251
Objective To discuss the efficacy of Compound Anisodine Injection combined with Iodized Lecithin Tablets in treatment of central serous chorioretinopathy.Methods Totally 60 patients with CSC were selected,and divided into two groups randomly.The patients in control group (29 cases) were given Iodized Lecithin Tablets.The patients in observation group (31 cases) were given Iodized Lecithin Tablets and Compound Anisodine Injection.The efficacy of Compound Anisodine Injection combined with Iodized Lecithin Tablets in treatment of central serous chorioretinopathy was evaluated by efficacy,visual acuity,light sensitivity,and adverse reaction during treatment.Results After treatment,the effective rates of observation and control groups were 93.5% and 79.3%,and the observation group was significantly higher than control group (P < 0.05).There were no statistical significance on visual acuity between two groups.After treatment,rhe visual acuity of two groups was increased and the visual acuity in the observation group was better (P < 0.05).Before treatment,there were no statistical significance on light sensitivity between two groups.After 2 and 4 weeks treatment,the light sensitivity of two groups were increased and the light sensitivity in the observation group was higher (P < 0.05).During treatment,there was no statistical significance on adverse reaction between two groups.Conclusion Compound Anisodine Injection combined with Iodized Lecithin Tablets has a curative effect on central serous chorioretinopathy.It could increase the visual acuity and improve the light sensitivity of eyes with good security.It is worthy of clinical use.
5.Sequence analysis and identification of a chloroplast matK gene in Rhei Rhizoma from different botanical origins.
Xiaoqin ZHANG ; Chunsheng LIU ; Xingli YAN ; Xiaoli CHENG ; Juan LIU ; Qiuling WANG ; Kai LIU ; Shengli WEI
Acta Pharmaceutica Sinica 2013;48(11):1722-8
Rhei Rhizoma is a Chinese medicine with multiple botanical origins. There is a problem to identify it with conventional methods. To compare the characteristics of chloroplast matK gene sequences of different Rheum species and authenticate inspected species, the matK gene sequences of different species from different origins were amplified, cloned, and sequenced. Genomic DNA of Rheum plants was extracted using modified DNA extracted Kit and matK gene sequences were analyzed by ContingExpress, DNAman and MEGA5.0. The length of matK gene sequences of Rheum palmatum, R. tanguticum and R. officinale were 1 518 bp containing 57 variable loci. According to the mutation sites, R. palmatum, R. tanguticum and R. officinale were divided into different genotypes separately. Based on the established method according to the loci 587, 707, 838, we successfully identified the genuine Rheum species from its adulterants.
6.Experimental study on brain injury in Beagle dogs caused by adjacent cabin explosion in warship
Yanteng LI ; Gang CHENG ; Shuai LIU ; Bangxin LIU ; Shuwei WANG ; Boyuan WEI ; Juan WANG ; Jianning ZHANG
Medical Journal of Chinese People's Liberation Army 2017;42(3):234-238
Objective Through the establishment of adjacent cabin blast injury model of Beagle dog,to investigate the pathophysiological changes in the experimental animals in this scenario,then speculate on the mechanisms of injury.Methods Several adjacent cabins were built in the same size with the real warship.Seven Beagle dogs were subjected to injuries from the explosion,from whom one was selected randomly to implant intracranial pressure transducers before blast,the others were tested on the pathophysiological changes after blast.The dogs were mounted on the platform of a cabinet in the adjacent cabin,subjected to injury from 650g bare TNT explosive blast.The transducers recorded the value of space and intracranial shock wave pressure.Following blast treatment,the serum levels of IL-6,IL-8,neuron specific enolase (NSE),brain and chest CT and pathological changes of the brain tissue were observed.Results Serum levels of IL-6,IL-8 and NSE were elevated to varying degrees after blast.All of them increased significantly at different time points after blast (P<0.05).Brain and chest CT examinations did not show any significant positive results.Pathological results showed that there was a little necrosis in the brain,some neurons had karyopycnosis,karyolysis or disappearance of the nucleoli,and the cell boundaries were blurred.The blast wave was blocked greatly by the scalp and skull (about 90%),but could still penetrate them and cause brain injuries.Conclusions Explosion in the adjacent cabin causes mainly mild traumatic brain injuries.Blast wave can be blocked by the scalp and skull greatly.
7.Effect of Yiqi Jianpi Herb on Content of NPY, VIP and Expression of Mapk14 mRNA of Rats with Spleen-qi Deficiency
Yongqiang DUAN ; Weidong CHENG ; Juan DU ; Xiaoyi YANG ; Liang LIU ; Zihan GONG ; Liming ZHU ; Yingxia CHENG ; Lanzhen LI
Chinese Journal of Information on Traditional Chinese Medicine 2014;(4):59-62,66
Objective To investigate the effect of Yiqi Jianpi herb on content of NPY, VIP and expression of Mapk14 mRNA of rats with spleen-qi deficiency. Methods Rats were randomly divided into normal groups, model group (observed on 7 d, 14 d and 21 d), treatment group of Yiqi Jianpi herb, 10 rats in each group. The spleen-qi deficient model rats were made by rhubarb, exhaustive and hungry method, and treatment group was treated with Sijunzi decoction (20 g/kg) for 21 d, then the content of NPY, VIP in serum and small intestine were evaluated with radioimmunoassay, and expression of Mapk14 mRNA in small intestine tissue was evaluated with real time fluorescent quantitative PCR. Results In model group, content of NPY was much lower than that of normal group (P<0.05), content of VIP and relative expression of Mapk14 mRNA in small intestine tissue were much higher than that of normal group (P<0.05), the difference was obvious in 21 d group. Compared with model 21 d group, content of NPY was increased (P<0.05), content of VIP and relative expression of Mapk14 mRNA in small intestine tissue were decreased (P<0.05) in treatment group. Conclusion Yiqi Jianpi herb has functions of adjusting NPY, VIP secretion and inhibiting abnormal expression of Mapk14 mRNA.
8.Rapid Determination of N-Methylcarbamoyl Adduct in Hemoglobin of Workers Exposed to N,N-Dimethylformamide by Ultra High Performance Liquid Chromatography-Mass Spectrometry
Chunmin WANG ; Qiang LIU ; Jian LI ; Bo XU ; Kai MI ; Juan CHENG
Chinese Journal of Analytical Chemistry 2014;(9):1326-1331
A rapid analytical method was developed for the determination of N-methylcarbamoyl adduct in Hemoglobin of workers exposed to N,N-dimethylformamide by ultra high performance liquid chromatography-mass spectrometry ( UPLC/MS/MS). About 0. 1 g of hemoglobin sample, 40 μmol/L of 3-methyl-5-isopropylhydantoin (MIH) as the internal standard and 4. 75 mL of HCl-acetic acid (2∶1, V/V) were added in the centrifuge tube, and mixed for 3 min. Then the tube was heated in boiling water bath for 1h. After cooling down, 200 μL of the mixture and 600 μL of formic acid-acetonitrile (1%) were added into 96-well extract plate. The vacuum pump pressure was controlled to make the sample collection elute within 2-4 min.The purified collection was transferred into the sample vial, and 3-methyl-5-isopropylhydantoin ( MVH ) as degration product of N-methylcarbamoyl adduct was quantified by UPLC/MS/MS in multiple reaction monitoring ( MRM ) by internal standard method. A good linear relationship was obtained in the MVH concentration range of 0 . 01-1 . 0 μmol/L with the correlation coefficient of 0 . 999 . The recovery of added MVH in the blank sample was 97 . 3% and the relative standard deviation was 1 . 7%. The limit of detection (LOD) was 0. 01 μmol/g. This method was proved to be fast and efficient.
9.Multiple measures to improve perioperative antimicrobial use in typeⅠin-cision operation
Juan LIU ; Zhuoping TIAN ; Shaoming ZHANG ; Chun CHENG ; Guoping CHEN ; Hong YU
Chinese Journal of Infection Control 2016;15(6):422-425
Objective To evaluate the effect of multiple intervention measures on perioperative antimicrobial use in pa-tients undergoing typeⅠincision operation in a hospital,and provide basis for rational use of antimicrobial agents. Methods 9 823 patients with type I incision operation in April and October of 2005-2012 were surveyed retrospectively, data of 2005 was as baseline,from 2006 to 2012,multiple measures,including training,examination,supervision,feed-back,and cooperation of relevant departments were conducted,antimicrobial use before and after intervention was com-pared.Results The qualified rate of perioperative antimicrobial use in patients undergoing typeⅠ incision operation in-creased from 14.20% in 2006 to 92.30% in 2012;the rate of combined use of antimicrobial agents was relatively higher (7.00%-9.00%)in 2006-2009,had a downward trend in 2010- 2012,and decreased to 3.20% in 2012.Types of an-timicrobial agents for prophylactic use in typeⅠincision operation in 2006 and 2007 were similar to that of 2005,the main used antimicrobial agents were cephalosporins,penicillin and it’s compounds,and aminoglycosides;the major antimicrobial prophylaxis in 2008-2012 were the first and second generation cephalosporins,penicillin and it’s compounds.Multivariate non-conditional logistic regression analysis showed that age (40 - 59 years old),departments (orthopedics,general surgery,and ophthalmology),and years (2011 and 2012)were the main influencing factors for the qualified use of antimicrobial agents(all P<0.05).Conclusion Implementation of multiple intervention measures can improve the qualified rate of perioperative antimicrobial use in typeⅠ incision operation,reduce types of antimicrobial use and rate of combined antimicrobial use.
10.The effect of the photon therapeutic apparatus on type 2 diabetic peripheral neuropathy
Xiao LIANG ; Zhiping LIU ; Qingfeng CHENG ; Juan YUAN ; Chengyun WU ; Qinghua ZHAO ; Qifu LI
Chongqing Medicine 2014;(6):656-657,660
Objective To observe the clinical efficacy of the photon therapeutic apparatus on type 2 diabetic patients with periph-eral neuropathy .Methods While giving conventional nerve nourishing ,improving microcirculation treatment and nursing ,adjuvant therapy with Carnation-11 photon therapeutic apparatus was applied to one lower limb only of the 20 diabetes mellitus patients with peripheral neuropathy and compare the curative effects by paired observation of the two lower limbs .Results The evaluation score of peripheral neuropathy dropped obviously with ankle reflex and thumb vibration sensation improved greatly after 10 days of treat-ment and remained an unchanged follow -up test 30 days later ,which means the clinical therapeutic effect of the treated limb was significantly superior to the comparison limb .The difference of the curative effects is statistically significant ( P < 0 .05 ) . Conclusion Conventional therapy accompanied by adjuvant treatment of photon therapy can relieve the clinical symptoms and phys-ical signs of diabetic patients with peripheral neuropathy ,which could improve curative effect on diabetic peripheral neuropathy .