Nature is abundant in protein scaffolds.By selecting suitable protein scaffold,display and screening methods,the rational and constrained random peptide library(RPL)can be constructed.Compared with the non-constrained RPL,it offered more opportunities for obtaining novel protein structures and more higher affinity ligands against the target molecules.At present,the protein scaffold constrained RPLs have been shown great potential in applications such as target selection,basic research,clinical diagnosis,medical therapy and so on.It is systematically introduced the structure bases,classification and construction of constrained RPL based on scaffolds,as well the recent great advances of application in selection against target molecules with S-S constrained scaffolds,antibodies,Zinc finger protein,Z domain,FN3 domain as important examples.

This study is to investigate the anti-allergic effect of anthocyanidin and to explore its possible mechanism. The experiments of passive cutaneous anaphylaxis reaction (PCA) and colorimetry were used to determine the effect of anthocyanidin on degranulation of mast cells in vivo. For in vitro study, various concentrations of anthocyanidin (100, 50 and 25 micromol x L(-1)) were added to the culture medium of mast cells cultured with 100 microg x L(-1) of dinitrophenyl (DNP) specific IgE overnight. The azelastine (100 micromol x L(-1)) was selected as the positive control. The antigen (DNP-human serum albumin, DNP-HAS)-induced release of degranulation was measured by enzymatic assay, histamine was determined by EIA, and interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-alpha) were measured by Western blotting, separately. In addition, the effects of anthocyanidin on phosphorylation of NF-kappaB, p38MAPK and Akt were observed by Western blotting. The results showed that treatments with anthocyanidin (100 and 50 mg x kg(-1)) were followed by a decrease in PCA of rats. Anthocyanidin (100 and 50 micromol x L(-1)) obviously suppressed the degranulation from mast cells, whereas results from anthocyanidin (100 and 50 micromol x L(-1)) group indicated significant inhibitory effect on histamine, the calcium uptake, TNF-alpha, IL-6, phosphorylation of NF-kappaB, p38MAPK and Akt of mast cells induced by antigen. Anthocyanidin may suppress the anaphylactic reaction by inhibiting the action of mast cells. NF-kappaB, p38MAPK and Akt at least in part contribute to this event.
Animals ; Anthocyanins ; pharmacology ; Anti-Allergic Agents ; pharmacology ; Calcium ; metabolism ; Cell Degranulation ; drug effects ; Histamine Release ; drug effects ; Immunoglobulin E ; immunology ; Interleukin-6 ; metabolism ; Male ; Mast Cells ; immunology ; metabolism ; physiology ; Passive Cutaneous Anaphylaxis ; drug effects ; Proto-Oncogene Proteins c-akt ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; Transcription Factor RelA ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism ; p38 Mitogen-Activated Protein Kinases ; metabolism

Female ; Humans ; Middle Aged ; Pharyngeal Neoplasms ; Pharynx ; Teratoma

Objective:To investigate frequencies of microsatellite instability(MSI)and loss of heterozygosity(LOH)in renal ceil carcinoma(RCC),and to discuss the relationship of clinicopathological characteristics of RCC with MSI and LOH. Methods:Twelve microsatellite markers located at chromosomes 3p,9p and 14q were selected to investigate microsatellite alterations(MSI and LOH)in 31 RCC specimens and their paired metastasis specimens by polymerase chain reaction- polyacrylamide gel elect rophoresis-ethylene dibromide(PCR-PAGE-EB)staining and sequencing.Results:The frequency of MSI could reached 61.3% and that of LOH could reach 54.8%.The highest frequency of MSI was at locus of D9S168(32.3%);the highest frequency of LOH was at locus of D3S1289(21.4%).No correlation was found between MSI or LOH and the patients' age,sex,pathology type and metastastis,except that MSI was correlated with TNM stage of RCC(P

Many Chinese drugs (CHD) have showed their significant effects of integral immune-regulation, and lots of researches have conducted in recent years for exploring their mechanism from different levels, like cytological, molecular and genetic levels. In this paper, the relation between immune-regulation of CHD and Toll-like receptors/nuclear factor-kappaB (TLRs/NF-kappaB) signaling pathway was introduced in brief based upon the achievements of previous researches. It was pointed out that the two are closely related, to explore mechanism of CHD in this way is meaningful not only for further deepening the theoretical understanding of CHD's pharmacological immunoregulation, but also be practically facilitate for enhancing therapeutic efficacy of CHD and developing new CHD.
Adjuvants, Immunologic ; pharmacology ; Drugs, Chinese Herbal ; pharmacology ; Humans ; NF-kappa B ; immunology ; Signal Transduction ; drug effects ; Toll-Like Receptors ; immunology

Objective To establish a method for isolation and cultivation of human invasive pituitary adenoma-derived fibroblast in vitro and explore their biological properties in order to investigate the role of the fibroblasts in the invasive growth of the pituitary adenoma.Methods The fibroblasts were isolated from hu- man invasive pituitary adenoma tissue by different rate of adhesion.Primary culture and passage culture were carried out,and growth characteristics of fibroblasts were observed under phase-contrast microscope in primary and passage culture.Immunohistochemical technique was used to identify the fibroblasts.The cell growth curve was measured by MTT method.The ultrastructure was observed under electronmicroscope.Results The cultured invasive pituitary adenoma-derived fibroblasts showed active proliferative ability.The cell bodies was bigger than other tissue fibroblasts,and the cell shape was irregular.The rough endoplasmic reticulum, Golgi complex and ribosome were profuse in the cytoplasm.The positive expression rate of collagen[and vi- mentin of the fibroblasts were above 95%.Conclusion The method of different rate of adhesion is success- fully utilized to culture human invasive pituitary adenoma-derived fibroblasts.These cells showed active prolif- erative ability,and maybe have an importent function in the invasive growth of the pituitary adenoma.

Adolescent ; Cholesteatoma ; complications ; Ear, External ; abnormalities ; Ear, Middle ; abnormalities ; Humans ; Male ; Temporal Bone

Objective To explore the influence of using lidocaine in the second stage of labor among primiparas. Methods 300 primiparas were divided into three groups. When Group A has uterus contractions with anal fold releasing, bilateral pudendal nerve was conducted by block anesthesia with lidocaine hydrochloride. When Group B has the uterus contractions with head visible on vulva gapping 2～3 cm, lidocaine hydroctdoride bilateral pudendal nerve was carried out by block anesthesia. Group C was not carried out any measures. Results The shortest time of the second stage is Group A. Group C is the longest one. Pain in groups A, B significantly reduced than in group C. The condition of perineum in Group A and B were better than that of in Group C, the rate of postero-lateral episiotomy and the level of perineal laceration in Group C were higher. Conclusions Bilateral pudendal nerve block anesthesia can shorten the second stage of labor and suturing time, alleviate the pain of delivery, reduce the rate of postero-lateral episiotomy and the perineal laceration, and raise the rate of perineal integrity. Compared with the head visible on vulva gapping, dilatation of cervix 10 cm can get the better effect of shortening the second stage of labor.

Virginiamycin acetyltransferase D (VatD) plays a vital rule in streptogramins resistance by chemically inactivating streptogramin A. Therefore, it is desirable to discover novel small molecular weight inhibitors of VatD via state-of-the-art virtual screening techniques. This "cocktail" strategy by combining VatD inhibitor with streptogramins may provide new therapeutic opportunity for resistant bacteria infections. Structure-based virtual screening method (molecular docking) was applied to rank and score a chemical database containing 300 000 commercially available compounds against the VatD substrate binding site. Twenty six out of the 200 top scored compounds from the docking calculation were selected and submitted to the VatD enzymatic inhibition assay. The plasmid pRSET B/vatD was constructed and transformed into E. coli (trxB) host cells for over-expression, and VatD enzyme was purified and validated by showing acetyltransferase activity to Virginiamycin M1. Three out of these 26 tested compounds showed enzymatic inhibition on VatD with IC50 168.6, 91.0 and 55.2 micromol x L(-1), separately. Other compounds could not be dissolved in the system and/or had little effect on the enzyme (IC50 > 200 micromol x L(-1)). To our knowledge, it is first time that small molecular weight organic compounds were identified as VatD inhibitors. It is expected that the VatD inhibitors identified at present study could serve as lead compounds for the further development of the novel therapeutic agents to overcome streptogramins resistance.
Acetyltransferases ; antagonists & inhibitors ; genetics ; metabolism ; Catalysis ; drug effects ; Drug Design ; Drug Resistance, Bacterial ; Enzyme Inhibitors ; chemistry ; metabolism ; pharmacology ; Escherichia coli ; genetics ; Genetic Vectors ; Kinetics ; Molecular Structure ; Plasmids ; Streptogramin Group A ; chemistry ; metabolism ; pharmacology ; Transformation, Genetic

Adolescent ; Adult ; Antigens, Nuclear ; metabolism ; Brain Diseases ; complications ; diagnosis ; pathology ; surgery ; Brain Neoplasms ; pathology ; Child ; Child, Preschool ; Diagnosis, Differential ; Epilepsy ; etiology ; Female ; Ganglioglioma ; pathology ; Glial Fibrillary Acidic Protein ; metabolism ; Humans ; Infant ; Magnetic Resonance Imaging ; Male ; Malformations of Cortical Development ; classification ; complications ; diagnosis ; pathology ; surgery ; Malformations of Cortical Development, Group I ; Microtubule-Associated Proteins ; metabolism ; Neoplasms, Neuroepithelial ; pathology ; Nerve Tissue Proteins ; metabolism ; Neurofilament Proteins ; metabolism ; Retrospective Studies ; Vimentin ; metabolism ; Young Adult