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.

Antigens, Tumor-Associated, Carbohydrate ; blood ; Female ; Ganoderma ; Gastrointestinal Neoplasms ; blood ; metabolism ; Humans ; Male ; Middle Aged ; Spores

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

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.

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

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.

AIM: To systematic review the effectiveness of refractive multifocal intraocular lens ( MIOL ) ReZOOM vs diffractive MIOL ReSTOR in the treatment of cataract.
METHODS: Randomized controlled trials comparing refractive MIOL ReZOOM with diffractive MIOL ReSTOR were identified by searching CENTRAL, MEDLINE, EMbase, WANFANG MED ONLINE, CMJD, SinoMed, and CNKI. Related journals also were hand-searched. Methodological quality of randomized controlled trials ( RCTs ) was evaluated by simple evaluate method that recommended by the Cochrane Collaboration. Data extracted by two reviewers with designed extraction form. RevMan software ( release 5. 2 ) was used for data management and analysis.
RESULTS:A total of 7 trials (846 eyes) were included for systematic review. Subgroup analyses were used according to different model comparison of ReSTOR MIOL. The results showed a significant difference in the mean of the best distance corrected intermediate visual acuity ( BDCIVA) in the ReZOOM MIOL group with WMD=-0.11, 95% CI (-0.16, -0.06) (P<0.0001). It showed a significantly difference in the mean of the uncorrected near visual acuity ( UCNVA ) , complete spectacle independent rate, halo rate and glare rate in the ReSTOR MIOL group with WMD= 0. 09, 95% CI (0. 05, 0. 14) (P<0-00001), WMD= 2. 62, 95%CI (1. 76, 3. 91) (P<0. 00001), WMD=1. 35, 95% CI (1. 15, 1. 60) (P=0. 0004) and WMD=1-29, 95% CI (1. 09, 1. 53) (P= 0. 003). There was no significant difference between the two groups in the mean of the uncorrected distance visual acuity ( UCDVA ) , the uncorrected intermediate visual acuity ( UCIVA) , the best corrected distance visual acuity ( BCDVA ) and the best distance corrected near visual acuity ( BDCNVA ) with WMD-0.03, 95% CI (-0.06, 0.01) (P=0.15), WMD= -0.04, 95% CI (-0. 09, 0. 01) (P=0. 10), WMD= -0. 01, 95%CI (-0. 04, 0-02) (P=0. 55) and WMD= 0. 06, 95% CI (-0. 06, 0. 17) (P=0. 32).
CONCLUSION: Patients implanted with ReZOOM MIOL can provide better BDCIVA; patients implanted with ReSTOR MIOL show better UCNVA, are less likely to appear light halo, glare and other visual adverse reactions; correction in spectacles cases, patients implanted with ReZOOM or ReSTOR MIOL have considerable performances in the far and near visual acuity.

AIM: To observe the effects of porous polyethylene ( Medpor) as a spacer graft in the reconstruction of large areas eyelid defect after the operation of malignant tumors of lower eyelids.
METHODS: Nineteen cases ( 19 eyes ) of malignant tumors of lower eyelid underwent the eyelid reconstruction were selected. Medpor lower eyelid inserts implantation were used to replace tarsal joint sliding conjunctival flap and pedicle flap, and repaired full -thickness lower eyelid defects then underwent eyelid reconstruction.
RESULTS: Appearance of eyelids and functional improvements were satisfactory with no stimulation on the eyeball and no effect on the visual function. Implants is with no absorption, shift, exclusion or infection and no tumor recurrence in all cases during the follow up for 6-36mo.
CONCLUSION: Medpor lower eyelid inserts implantation can instead tarsal plate for the reconstruction of medium to large areas lower eyelid defect, which is easy performing with rare complications. It is an ideal alternatives of tarsal plate.

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

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.