The cytolytic effect of perforin is a mechanism of anti-virus,killing microbial-infected cells and tumor cells.Perforin is a very important non-specific immune factors in fish.In order to understand the function of perforin,the cDNA of grass carp perforin C-terminal peptide was amplified from grass carp liver and kidney cDNA library.It contains a protein kinase C conserved region 2(C2).The cDNA was connected with pET32a,and transformed to expression bacteria DE3.PFP-C was expressed by a prokaryotic expression system and then purified by affinity chromatography.It showed a significant haemolytic activity when tested with rabbit red cells,the optimal pH for haemolytic activity was 7.5,and its haemolytic function dependents on Ca2+ apparently.

Chronic Disease ; Graft Rejection ; therapy ; Humans ; Integrative Medicine ; Kidney Diseases ; etiology ; therapy ; Kidney Transplantation ; adverse effects

Botulism ; therapy ; Child ; Female ; Humans ; Respiration, Artificial

Objective:To identify the function of extracellular soluble vimentin that promotes proliferation, activation and chemotaxis of inflammatory cells. Methods: The proliferation of rat splenocytes stimulated with vimentin was evaluated in vitro. The lymphocyte counts and vimentin-antibody levels of peripheral blood in mice immunized with vimentin were detected in vivo. Peritoneal macrophages were collected and cultured with different concentrations of vimentin to detect the effect on phagocytosing chicken red blood cells. The chemotaxis of NIH3T3 fibroblast towards vimentin was observed in transwell chamber. Results:In vitro vimentin dose dependently promoted the proliferation of splenocytes. The proliferation indexes of primed and naive splenocytes cultured with 16μg/ml vimentin were reached up to ( 196. 0 ± 9. 7 )% and ( 208. 9 ± 4. 6 )% respectively without significant difference. In vivo vimentin significantly enhanced the lymphocytes number(109 L-1)of peripheral blood(5. 74±0. 51 vs. 1. 69±0. 13)and the levels of vimentin specific antibody( OD value 2. 31 ± 0. 06 vs. 0. 19 ± 0. 08 ) that shown no significant difference from immunization with vimentin plus CFA. In vitro vimentin dose dependently stimulated phagocytic ability of macrophages and performed the chemotactic effects on NIH3T3 fibroblasts. Conclusion:Extracellular soluble vimentin promotes the proliferation,activation and chemotaxis of concerned inflammatory cells and possesses the functions as an alarmin.

Chlorine ; Computer Simulation ; Computers, Analog ; Diffusion ; Software

Objective:To evaluate the efficacy of Epstein-Barr nuclear antigen 1/immunoglobulin A (EBNA1/IgA), BamH1 Z transactivator/IgA (Zta/IgA), capsid antigen/IgA (VCA/IgA), and Epstein-Barr virus deoxyribonucleic acid (EBV-DNA) in detecting different stages of na-sopharyngeal carcinoma (NPC). The relationship between the EBV markers and stages of NPC was also analyzed. Methods:Blood sam-ples of 152 untreated patients with NPC and 675 healthy subjects were collected.ELISA was used to detect the serum levels of EBNA1/IgA, Zta/IgA, and VCA/IgA. Fluorescence quantitative PCR (FQ-PCR) was used to detect the plasma levels of EBV-DNA. ROC and correla-tion analyses were employed to assess the detection assays for NPC diagnosis. The positive rates of EBV markers in NPC patients in dif-ferent stages were analyzed statistically. Results: The positive rates of EBNA1/IgA, Zta/IgA, VCA/IgA, and EBV-DNA in NPC patients were higher than those in the healthy individuals. The expression of EBNA1/IgA was relatively high in early NPC. The sensitivity of EB-NA1/IgA was 77.8%. In advanced NPC, the level of EBV-DNA was high, and the sensitivity of EBV-DNA was 88.8%. The specificity of EBV-DNA and EBNA1/IgA could reach more than 96%. The combination of EBV-DNA and EBNA1/IgA showed the best diagnostic value, with a sensitivity of 92.1%(early stage 82.5%, advanced stage 98.9%) and a specificity of 96.9%. The positive rates of EBV-DNA were positively associated with the NPC clinic stage and N stage. The positives rates of Zta/IgA were positively associated with the NPC N stage. Conclusion:The best single index for NPC screening in an asymptomatic population is EBNA1/IgA. EBV–DNA is an ideal index for auxiliary diagnostics of advanced NPC. The combination of EBV-DNA and EBNA1/IgA shows the best diagnostic value. EBV-DNA is an important index in the stage and illness monitoring of NPC. Zta/IgA can indirectly reflect the character of lymph node metastasis, and it may be useful in assessment of NPC surveillance.

Objective To observe the effect of Yishen Paizhuo Decoction(Decoction for reinforcing the kidney and discharging the turbid)on chronic renal failure(CRF)patients.Methods The 82 CRF patients were randomized into observation group(42 cases) and control group(40 cases).In addition to the routine treatment,the observation group was administered modified Yishen Paizbuo Decoction,and the control group was given coated aldehyde oxystarch.The course of treatment was 6 months for both groups. Changes of clinical symptoms,renal functions,blood fat,and hemoglobin were observed,and the linear regression evaluation and renal survival evaluation were employed to evaluate the development of CRF.Results The improvement of observation group in renal func- tions and blood fat was remarkable with the increased renal survival rate,compared with the control group,the difference was signifi- cant(P

Objective To analyze the expression of phosphorylated protein kinase B(pAKT)and PTEN protein in ovarian epithelial cancer and to investigate the correlations between their expression and prognosis of ovarian epithelial cancers.Methods Expression of pAKT and PTEN in 12 normal ovarian tissues,20 benign tumors,12 borderline tumors and 80 cases of ovarian epithelial cancers were detected by immunohistochemical method,and their correlations were analyzed.Results The positive expression of pAKT in normal ovarian and benign tumor tissues were significantly lower than that in ovarian epithelial cancers(8%,10% vs 55%;P

Lycii Cortex, a popular herb medicine in traditional Chinese medicine, is used to treat different inflammation-related diseases. The aim of our work is to find the key constituents inhibiting NF-kappaB, a key regulator of inflammation. In the investigations of cell-based in vitro assays of extracts, we found that both ethyl acetate extract and methanol extract of Lycii Cortex inhibited the TNF-alpha-induced activation of NF-kappaB. Through bioassay-guided fractionation, we identified 4 phenolic amides including trans-N-(p-coumaroyl) tyramine (1), trans-N-feruloyltyramine (2), trans-N-caffeoyltyramine (3), and dihydro-N-caffeoyltyramine (4). Four phenolic amides showed differently inhibitory activities on TNF-alpha-induced NF-kappaB activation. Trans-N-caffeoyltyramine (3) was identified as the key component with an IC50 of 18.41 micromol x L(-1). It was suggested that the hydroxyl group at C-3 in trans-N-caffeoyltyramine might be a key binding site and its C-7,8-double bond might play an important role on NF-kappaB inhibitory activities as the link of the conjugation of pi electrons leading to a partial planar conformation. It might be inferred that the biological activity of compound 3 is attributed to the structure of Michael reaction acceptor containing alpha, beta-unsaturated ketones and benzene along with hydroxyl group in o-diphenol.
Biological Assay ; Cell Line ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; pharmacology ; Humans ; Inflammation Mediators ; antagonists & inhibitors ; immunology ; Lycium ; chemistry ; Molecular Structure ; NF-kappa B ; antagonists & inhibitors ; immunology

To obtain the genome sequence of human bocavirus 2 (HBoV2), different regions of HBoV2 genome were amplified through PCR in fecal specimens which had been identified as single-positive for HBoV2 in 2010. A genome sequence of HBoV2 (HBoV2-NC, 5444 bp) was obtained after sequence assembly. The phylogenetic analysis showed that HBoV2-NC had the closest evolutionary relationship with HBoV2 Lanzhou strain. The predication of inverted terminal repeats of HBoV2-NC by DINAMelt showed that inverted terminal repeats were contained in HBoV2-NC 5' terminal, which had the typical stem-loop structure in other parvoviruses. Finally, some flanking sequences of HBoV2-NC were amplified by linker-PCR.
Base Sequence ; Gene Amplification ; Genome, Viral ; Human bocavirus ; chemistry ; classification ; genetics ; Humans ; Molecular Sequence Data ; Nucleic Acid Conformation ; Parvoviridae Infections ; virology ; Phylogeny ; RNA, Viral ; chemistry ; genetics ; Terminal Repeat Sequences