2.Cloning of anti-MMP-2 human antibodies from semisynthetic phage antibody library
Xiuyun DING ; Yan WANG ; Bing HUA ; Al ET
Chinese Journal of Immunology 2000;0(08):-
Objective:To clone human anti MMP 2 antibodies from semisynthetic phage antibody library.Methods:Panning of semisynthetic phage antibody library against recombinant human MMP 2 was conducted to select specific antibodies.The antigen binding characterastics were analyzed by ELISAs.Results:MMP 2 binding phage antibody clones were obtanied after four rounds of panning,but they all showed binding activity to unrelated ags tested.One clone,AD20,was chosen for further analysis by competitive ELISA.It was found that the binding of AD20 to MMP 2 could be inhibited only by free MMP 2 but not unrelated Ags,while the binding to other Ags could not be inhibited by Ags tested,including MMP 2.Neutralization test demonstrated that AD20 could neutralize the enzymatic activity of MMP 2.Conclusion:A neutralizing human antibody against MMP 2 was obtained from a semisynthetic phage antibody library,which shows bindings with unrelated Ags nonspecifically that may be caused by different binding modes,a phenomenon termed polyreactivity.
3.Experimental study on relationship between pungent-hot herb property express and calmodulin.
Xin LIU ; Yan HU ; Yi-Ran CUI ; Xiao-Qing LIU ; Bing-Hua TANG ; Bing ZHANG
China Journal of Chinese Materia Medica 2014;39(5):873-878
OBJECTIVETo explain the essence of pungent-hot herb property express according to in vivo and in vitro studies on its effect on calmodulin on the base of the observation of the adjustment in hypothalamic-pituitary-gonad axis functions of Aconiti Lateralis Radix Praeparata, Curculiginis Rhizoma, Cinnamomi Cortex and bitter-cold herb Phellodendri Chinensis Cortex in rats under the state of yang deficiency.
METHODThe yang-deficient model was duplicated by intramuscularly injecting hydrocortisone sodium succinate powder injection. After the intervention with Aconiti Lateralis Radix Praeparata, Curculiginis Rhizoma, Cinnamomi Cortex and bitter-cold herb Phellodendri Chinensis Cortex for seven days, TSH, T3, T4, 17-OHCS, COR, T, E2 of hypothalamus-pituitary-target gland axis and other relevant indexes were detected. The calmodulin expression in livers and L02 cells cultured in vitro was detected by using Western blot.
RESULTPungent-hot herbs Aconiti Lateralis Radix Praeparata, Curculiginis Rhizoma, Cinnamomi Cortex can significantly correct indicators of hypothalamic-pituitary-gonad axis and calmodulin, whereas the bitter-cold herb Phellodendri Chinensis Cortex showed no obvious effect.
CONCLUSIONThe pungent-hot herb property expression may be closely related to calmodulin.
Animals ; Calmodulin ; metabolism ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; Gonads ; drug effects ; metabolism ; Hypothalamo-Hypophyseal System ; drug effects ; metabolism ; Liver ; drug effects ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley ; Yang Deficiency ; drug therapy ; metabolism
4.Cloning,Expressing and Sequence Analysis of a Novel vip3Aa Gene
Jun CAI ; Dong-Hui HUA ; Liang XIAO ; Bing YAN ; Yue-Hua CHEN ;
Microbiology 1992;0(04):-
The vip3A gene of Bt9816C was cloned and the sequencing result was submitted to GenBank (accession no.AY945939). The gene was identified as a novel vip3Aa gene, which was assigned name vip3Aa18 by the Bacillus thuringiensis delta-endotoxin nomenclature committee. Subsequently, vip3Aa18 was expressed in Escherichia coli BL21 and bioassay demonstrated that the purified recombinant Vip3Aa18 had high toxicity against Helicoverpa armigera and Spodoptera exigua. The results of sequence analysis revealed that a carbohydrate binding domain exists on the C-termini 536 to 667 residues of Vip3Aa18,which maybe participate in binding to midgut receptors in susceptible insects. Moreover, a transmembrane helices located on N-termini 272 to 292 residues was proposed responding for pore formation. Furthermore, a putative disulfide bond was found in the Vip3Aa18 sequence. The specific structures and sites of Vip3Aa18 sequence imply potential function.
5.Application of adrenocorticotropic hormone receptor determination and ultrastructural observation of tumor cells in the subtyping of adrenocortical neoplasms.
Hua FAN ; Han-Zhong LI ; Wei-Gang YAN ; Bing-Bing SHI
Acta Academiae Medicinae Sinicae 2011;33(6):679-684
OBJECTIVETo explore the values of adrenocorticotropic hormone receptor (ACTH-R) determination and ultrastructural observation of tumor cells in the subtyping of adrenocortical neoplasms (ANs).
METHODSThe expression of ACTH-R in 87 AN tissues were determined with Polymer immunohistochemical staining, with 10 normal adrenal tissues as the controls. The ultrastructure of the tumor cells was observed using electron microscopy.
RESULTSThe positive expression rate of ACTH-R was (80.1 +/- 8.2)%, (53.2 +/- 10.3)%, (63.2 +/- 10.1)%, (83.3 +/- 6.5)%, and (70.1 +/- 7.3)% in the sub-CPA group, CPA group, APA group, NFA group, and NC group, respectively. ACTH-R expression was significantly higher in NFA and sub-CPA groups than in NC group (P = 0.001, P = 0.000), APA group (P = 0.000, P = 0.000), and CPA group (P = 0.000, P = 0.000), and was also significantly different between NC group and APA group (P = 0.039) and between APA group and CPA group (P = 0.037). However, no significant difference was found between NFA group and sub-CPA group (P = 0.325). As shown by the electron microscopy, ANs had some partially similar microscopic features, while different AN subtypes showed differences in the type and amount of secretory granules.
CONCLUSIONACTH-R determination and ultrastructural observation of tumor cells may be helpful for subtyping ANs.
Adrenal Cortex Neoplasms ; diagnosis ; metabolism ; ultrastructure ; Adrenal Glands ; metabolism ; ultrastructure ; Adult ; Female ; Humans ; Male ; Middle Aged ; Receptors, Corticotropin ; metabolism
6.Study on physical and mental health status and influencing factors of middle-aged intellectuals.
Xin-hua LI ; Bing DENG ; Yan ZHU
Chinese Journal of Epidemiology 2003;24(9):778-782
OBJECTIVETo understand physical and mental health status of middle-aged intellectuals and related risk factors on mental health under the present social conditions.
METHODSStratified-cluster sampling and questionnaire survey were used. One thousand five hundred and three middle-aged intellectuals were investigated by SCL-90 test and influential reasons, using general relative risk model. Effects and interaction modes of risk factors were analyzed.
RESULTSThe prevalence rate of middle-aged intellectuals was 241.5 per thousand per two weeks, while the prevalence rate of chronic disease was 306.7 per thousand. 6.9% of the middle-aged intellectuals (male 6.1%, female 7.8%) had mental health problems. Results suggested that the major risk factors were, as listed in a order of their effects on mental health disorders: divorced or losing spouse, exposure to poison and noisiness, lack of hobbies, long working hours, having chronic diseases.
CONCLUSIONPhysical and mental health status of middle-aged intellectuals were under by many kinds of pressures. Multiple measures should be taken to improve the health status of the middle-aged intellectuals.
Adult ; Chronic Disease ; Female ; Health Status ; Humans ; Male ; Mental Health ; Middle Aged ; Multivariate Analysis ; Risk Factors
7.Acupuncture at Shangjuxu (ST37) to Inhibit Distention of Colon Induced Discharge Reaction of LC: an Experimental Study.
Qing-yan ZHU ; Hua WANG ; Ze-bing CHEN ; Xi-quan LIU ; Xin JIN
Chinese Journal of Integrated Traditional and Western Medicine 2015;35(5):608-611
OBJECTIVETo study possible mechanisms of Shangjuxu (ST37) and the large intestine.
METHODSTotally 40 SD rats were selected. The distension of end colon was used as injured afferent stimulus. Activities of locus coeruleus (LC) neurons were recorded by extracellular microelectrode technique. Shangjuxu (ST37) and Hegu (L14) were needled to observe general features of discharge reactions, distention of colon induced discharge reactions of LC, and its effects on distention of colon induced discharge reactions of LC.
RESULTSDistention of colon could induce incrased discharge of LC neurons by 127.33% ± 45.48%. But needling at Shangjuxu (ST37) and Hegu (L14) could inhibit this injured response by 38.24% ± 7.69% and 21.29% ± 13.16% respectively (all P < 0.01).
CONCLUSIONSNeedling at Shangjuxu (ST37) and afferent signals of colon distension converged and interacted with each other. Needling at Shangjuxu (ST37) could significantly inhibit colon distension induced discharge of LC neurons, which might be one of mechanisms for Shangjuxu (ST37) and the large intestine relationship.
Acupuncture Therapy ; Animals ; Colon ; Intestine, Large ; Locus Coeruleus ; physiology ; Neurons ; Rats ; Rats, Sprague-Dawley
8.Acrylic resin bone cement composite as a bone substitute for subchondral bone induces knee joint osteoarthritis
Hua JIANG ; Yu YAN ; Hongbing MA ; Bing XU ; Junrui WANG ; Dianming JIANG
Chinese Journal of Tissue Engineering Research 2015;(25):3962-3966
BACKGROUND:The hardened acrylic resin bone cement composite after implantation into human body can resist an intensity of 78-93 MPa. But a large amount of heat energy is released by bone cement during the process of solidification and it wil kil normal cel s, leading to peripheral tissue necrosis. OBJECTIVE:To investigate the characteristics of induced knee joint osteoarthritis after application of acrylic resin bone cement composite as a bone substitute for subchondral bone. METHODS:Thirty male Japanese big ear rabbits were randomly and equal y divided into four experimental groups (A, B, C, D) and a blank control group. After removal of subchondral bone on the right medial tibial plateau, polymethyl methacrylate powder/hydroxyapatite composite materials were implanted in rabbits in the experimental groups A, B, C, and D. Rabbits in the blank control group were only subjected to exposure of periosteum on the left medial tibial plateau. At 3, 6, 9 and 12 weeks after removal of subchondral bone, rabbits in the experimental groups A, B, C and D were sacrificed, and subchondral bone specimens were taken for hematoxylin-eosin staining and matrix metal oproteinase expression analysis. At the same time, interleukin-1βand tumor necrosis factor-ɑ levels in the synovial fluid were determined. RESULTS AND CONCLUSION:Mankin score in the experimental group C was significantly higher than in the blank control group and experimental group A (P<0.05). Mankin score in the experimental group D was significantly higher than in the experimental group B (P<0.05). The gray scale of matrix metal oproteinase-1 was highest in the blank control group, fol owed by experimental groups A, B, C, and the last in the experimental group D (P<0.05). Interleukin-1β and tumor necrosis factor-ɑ levels in the synovial fluid were highest in the experimental group D, fol owed by experimental groups C, B, A and the last in the blank control group (P<0.05). These findings suggest that acrylic resin bone cement composite as a bone substitute for subchondral bone induces knee joint osteoarthritis and leads to increases in matrix metal oproteinase-1 and tumor necrosis factor-ɑ levels in the synovial fluid.
9.Establishment of large phage antibody library
Yuping CHEN ; Guomin ZHANG ; Yuanyuan QIAO ; Gang WANG ; Yufeng LIU ; Bing HUA ; Yan WANG
Chinese Journal of Immunology 1985;0(05):-
Objective:To create a large human phage antibody library from which easy to get diabody and select human antibody clones.Methods:VL and VH genes were amplified by RT-PCR from RNA which came from normal adult peripheral blood and new-born cord blood lymphocytes. VL and VH genes were reamplified to add a region of overlap in the ScFv linker to form ScFv genes in which VH genes were flanked by two non-homologous loxp sites. The ScFv genes were cloned into PDF to obtain a primary library. This primary library was used to infect bacteria Bs1365(expressing cre recombinase) with high multiplicity of infection(MOI) 200∶1. This procedure resulted in a very large phage antibody library through VL and VH recombined by the cre recombinase. Following recombination, phagemide were derived from these bacteria and used to infect bacteria not expressing cre(XL1-Blue) at MOI
10.Application of bandage contact lens in pterygium excision combined with conjunctival transplantation
Zheng-Hua, XU ; Xiao-Ling, ZHANG ; Yi, LIU ; Xiao-Wei, GAO ; Bing, REN ; Yan, CAI
International Eye Science 2017;17(9):1786-1789
AIM:To evaluate the application and effect of bandage contact lens in pterygium excision combined with conjunctival transplantation (CAT).METHODS:In a prospective, randomized, controlled clinical study, 110 patients (110 eyes) diagnosed with primary pterygium were collected in PLA NO.474 Hospital from January 2015 to January 2016.The 110 patients enrolled in the study in turn, and divided into two groups by the odd and even number.The odd number divided into bandage contact lens group (CAT + bandage contact lens, n=55), while the even number divided into control group (CAT, n=55).Visual analog scale (VAS) and corneal irritation were evaluated on 1, 3 and 7d after operation.Cornea fluorescent staining testing was carried out on 3d after operation.Following-up all the patients with 1a at least observed the recurrence of pterygium.RESULTS:The score of VAS in bandage contact lens group less than that in control group on 1d (4.13±2.06 vs 5.80±1.93, t=4.391, P<0.001) and 3d (2.09±1.36 vs 3.65±1.65, t=5.422, P<0.001) after operation, while there was no significant difference between two groups on 7d (t=1.295, P=0.198) after operation.The corneal irritation in bandage contact lens group less than that in control group on 1d and 3d after operation (P<0.05), while there was no significant difference between two groups on 7d after operation (P=0.052).Cornea fluorescent staining testing area in bandage contact lens group was less than that in control group on 7d after operation (0.33±0.37mm2 vs 2.73±2.21mm2, t=7.921, P<0.01).There was no significant difference in recurrence rate between two groups after 1a operation (P=1.000).CONCLUSION:Bandage contact lens could significantly release pain and corneal irritation, promote the healing of the corneal epitheliums in the defected area, and increase the postoperative comfort level in patients after operation.